We are beginning to look at replacements for our aging (8+ yrs) Anatech Hummer VII. This coater is a bench top unit used for gold coating only although we do have the carbon coater attachment. This is a standard mechanical pump unit (non-turbo).
Our work is all materials related (non-biological) and we typically examine specimens in the 50X to 10,000X region, with an extremely varied selection of parts and contaminants. The SEM is a LaB6, so I don't believe I need the ability to deposit chromium or other metals used for high magnification work.
The main features I would look for are bench-top size; relatively quick cycle times; automatic push-button operation, with the ability to have some manual control for bleeding in argon to help dry samples; and a cost in the $8-10K region. The ability to effectively carbon coat samples would be nice if it worked well. The current Hummer unit has problems when trying to carbon coat filter pads.
I would like to solicit input from the group on their recommendations for purchase of a new coater and any experiences with reliability issues of the various units that are available.
Thanks in advance for any help.
John Giles Senior Materials Engineer Honeywell Space Systems
Thanks to everyone who shared their experience with long term fixation. Before any field work is started, we will use HEPES and PIPES buffers with the GTA fixative and compare the results after storing mammalian brain tissue for -at- 1month in both GTA/buffer and buffer alone. I don't know when this will occur, as it depends on grant applications, coordination with other labs, etc. If anyone is interested in what we discover with this comparison, please send me an e-mail message and I will contact you with the results. thanks again, Louisa Howard EM Facility-Remsen 240 Dartmouth College Hanover, H.N. 03755
While following the thread on pump rebuilds, I wondered if some of you have extended experience with the ages of roughing pumps? What are the ages typically achieved with the usual routine maintenance? I recently had two Edwards pumps fail on me. The cause seemed to be excessive water build-up in the oil chamber that caused rusting. These pumps are used only to prepump film desiccaters. I also found that some of the Al parts were corroded, possibly from calcium sulfate dust from the desiccant. I run the pumps with the ballast somewhat open, but it seems that the exhausted water condenses in the hoses we use to vent the fumes and drips back down into the pump housing. I am wondering if my pumps are prematurely failing or if they're reaching a ripe age (one is about 15 years old).
Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets The Johns Hopkins University Baltimore, Maryland 21218
Message-Id: {2.2.16.19960501190617.223f4c5a-at-email.uncc.edu} X-Sender: sfzane-at-email.uncc.edu X-Mailer: Windows Eudora Pro Version 2.2 (16) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Hello fellow microscopists, On the 17th of April I sent out a questionnaire regarding the administrative hierarchy of your labs. There was a very healthy response to this questionnaire and I have attempted to summarize the responses for those of you who might be interested. The one area which I haven't yet been able to deal with yet is the various responsibilities. So, what I will do is post what I have and when I find time to deal with the rest, I will try to post that,too. I really do appreciate all your comments. They have been very helpful. For those who asked, I am making an attempt to revitalize our lab. We have a TEM and all ancillary equipment. And we have access to a SEM and have in our lab a sputter coater, a critical point dryer and a vacuum evaporator. As the EM work has dropped off in recent years, I have proposed that we form an integrated microscopy lab. One of the first additions I would like to see is a confocal microscope. There are also a couple of fluorescence microscopes in the department as well as some video equipment which might, (or not) become a part of the integrated facility. However, I am the only person employed in the lab at the present time. I do everything from washing glassware to producing the micrographs of the EM projects, ordering, scheduling, etc. Any students or others (some from outside the university community) needing assistance come to me. So I thought that before I bit off more than I could chew, I would ask you how you ran things. Each and every response which I received was helpful in one way or another and I really do appreciate your taking time to help me out. Following is the portion of the summary which I have completed. There were 25 responses and out of those 25:
16 have directors and 9 have coordinators (lab managers or lab supervisors)
Of the 16 directors, 12 are faculty and 4 are staff employees.
Of the 12 faculty directors, 10 have 12 month contracts. The 4 staff directors also have 12 month contracts.
Of the 9 coordinators their supervisors ranged from:
1. senior staff committee, 2. nominated academic staff, 3. dean of COAS, 4. dept. head, 5. faculty committee.
All have 12 month contracts. All are staff except 1 who is a bit unclear.
Again, thank you very much.
Best wishes, Sandra Sandra F. Zane, EM Tech. sfzane-at-email.uncc.edu Dept. of Biology, UNCC Ph.(704)547-4051 9201 University City Blvd. Fax (704)547-3128 Charlotte, NC 28223
We are beginning to look at replacements for our aging (8+ yrs) Anatech Hummer VII. This coater is a bench top unit used for gold coating only although we do have the carbon coater attachment. This is a standard mechanical pump unit (non-turbo).
Our work is all materials related (non-biological) and we typically examine specimens in the 50X to 10,000X region, with an extremely varied selection of parts and contaminants. The SEM is a LaB6, so I don't believe I need the ability to deposit chromium or other metals used for high magnification work.
The main features I would look for are bench-top size; relatively quick cycle times; automatic push-button operation, with the ability to have some manual control for bleeding in argon to help dry samples; and a cost in the $8-10K region. The ability to effectively carbon coat samples would be nice if it worked well. The current Hummer unit has problems when trying to carbon coat filter pads.
I would like to solicit input from the group on their recommendations for purchase of a new coater and any experiences with reliability issues of the various units that are available.
Thanks in advance for any help.
John Giles Senior Materials Engineer Honeywell Space Systems
On Wed, 01 May 1996 16:27:24 -0400 (EDT) Kenneth JT Livi wrote
} What are the ages } typically achieved with the usual routine maintenance?
These comments refer to both belt and direct drive pumps. Life is about 10 - 15 years, and longer on cleaner systems
} I recently had two Edwards pumps fail on me. The cause seemed to be } excessive water build-up in the oil chamber that caused rusting. These } pumps are used only to prepump film desiccaters.
Very hard on the pumps and you could expect shortened life, I think
} the Al parts were corroded, possibly from calcium sulfate dust from the } desiccant. I run the pumps with the ballast somewhat open, but it seems } that the exhausted water condenses in the hoses we use to vent the fumes } and drips back down into the pump housing.
We have always used a sump in the exhaust line as close as possible to the pump itself and it is amazing the amount of fluid which is collected.
} I am wondering if my pumps are prematurely failing or if they're reaching a } ripe age (one is about 15 years old).
I think that's probably pretty good. Cheers jjm Professor John J. Millar, PhD Department of Applied Physics and Electron Microscope Unit RMIT, GPO Box 2476V, MELBOURNE AUSTRALIA 3001 +613 9660 2602 fax 613 9660 5290 email jjmill-at-rmit.edu.au
} I am wondering if my pumps are prematurely failing or if they're reaching a } ripe age (one is about 15 years old). } } Ciao for now, } Ken
There are several practices applicable to your case:
If the moisture is chronic and the use is intermittent, motoor oil. It is amazing stuff, and will emulsify the H2O, loosing only a little VP due to additives.
The heavy duty H2O applications with continuous operation use a heavier oil weight which causes the pump to run hotter than 212F.
GeoffA, For 3D reconstruction as well as digital confocal (i.e., reduction of out-of-focus haze in optical sectioning) VAYTEK, Inc. has several very nice software products for PC. Their software is available as well for Mac and for UNIX systems. VOXBLAST will do your 3D reconstruction. I understand that VAYTEK has developed a new product for "rubber sheeting" sections for a better fit, though I've not seen this. MicroTome and HazeBuster are products that employ digital deconvolution methods to sharpen each image in a series of frames acquired at incremental planes of focus in a thick section. The deconvolution algorithms are much more sophisticated than conventional edge enhancement filters found most image processing software. The VAYTEK products are available as "stand-alone" windows programs or as "plug-ins" to image analysis packages such as Media Cybernetics Image Pro Plus. You can get more information about the products at VAYTEK's web site http://www.vaytek.com
Good luck!
John Libert OPELCO OPtical ELements COrporation
GeoffA wrote: } } Hi all from Sydney where, I'm sad to say, Winter is on the way and it's } a wet sub-20 degrees. And I note that all those conferences in } Australia have affected Nestor when he opens a message with "G'day"! } Will he trade that hat of his for a Croc Dundee model? } } One of our colleagues would like to do 3D reconstructions from serial } histological sections on a PC. Over the years he has tried this a few } times with drawing tablets and various early graphics software but } always given up because it has been ultimately easier to do it with } pencil and paper. He now has a better setup with 'scope, CCD and speedy } Pentium but we still have the perennial problem of registering } successive sections. } } Can anybody tell me of any DOS/Windows (even Unix) image analysis } software (or dedicated reconstruction software) which allows for } independent registration of slices within a stack and maybe } 'rubber-sheeting' as well? } } I know that Prism allows this but it is Mac-based. Elsewhere in our } Museum we have Mac versions of both Dicer and VoxelView (around 1990 } versions) but neither of these lets us move sections. } } Any help gratefully received, } } Geoff Avern } Microscopy Laboratories } Australian Museum } Sydney, Australia.
Besides Tech no Trade (who has yet to send me a new phone number, catalog, address) where might I get platinum pellets, hollow carbon, and filaments for my BAF 301? I have talked to a company that will help me out with the carbon and platinum, but the tungston filaments are another story, or maybe techno trade can get me their number so I can throw a couple of hundred bucks their way.
John Grazul Rutgers University Electron Microscope Facility
The point source bulbs for the L1200 are called PULAM, they are custom made and cost $128 each. These are 24V 100Watt bulbs.
For older units like S45 and S138, S183, there is an adapter for the Edison base (usual lamp socket) to adapt to a bayonet bulb. These are not as good but the only thing that fits the physical constraints in the old units. The adapter is $60 and the bulbs are $12. These are 20Volt 100 Watt bulbs.
We offer an educational discount of 8%.
Please make sure on the old enlargers that you have removed the heat filter normally used only with the 200-300 watt diffuse bulbs, they are almost surely fogged with age and totally eliminate the benefits of point source. The normal voltage for the transformer in the old units is 110 volts, you must make a voltage adapter.
For other questions please feel free to call us.
Steve Miller Integrated Microsytems, Inc. (An Authorized Durst Dealer) P.O.Box 1074 Park Ridge, IL 60068 Phone 800-388-8801 Fax 847-696-2541
Please make your dinner reservations today (May 2) or tomorrow for this event.
} Northern CA Society for Microscopy (NCSM) Meeting Announcement } } Where: Roche Biochemicals (formerly Syntex), Gallery } Conference Center Bldg A-2 } 3401 Hillview Ave, Palo Alto CA } } When: Thursday, May 9, 1994, from 2:30 to 8:45 PM } } } 2:30 -3:00 Registration } Speakers } 3:00-3:45 Hank W. Bass, UC Berkeley, Molecular and Cellular } Biology, "Analysis of Meiosis using 3-D } Deconvolution Light Microscopy". 4:00-4:45 David Blake, } NASA Ames Research Center, Space Sciences Division } "Electron Microscopy of Astrophysical Ice" } 5:00 - 6:30 Social Hour and Local Society A filiate meeting } sponsored by JEOL Instruments } 6:30-7:30 Dinner. Choices: Marinated Stripped Loin with sundried } tomatoes, Lemon chicken with caper sauce, or roasted } vegetable lasagna, } each with garlic mashed potatoes, summer squash medley, } mixed green salad, dinner rolls, and dessert. } 7:45 - 8:45 Paul Carpenter, Manager Analytical and Geology, Cal } Institute of Technology, "XRay and Optical } Methods as Applied to the Study of Gemstones." } } Please help us to plan accurately. Make your reservation before Friday MAY } 3rd } Fax reservation(s) with meal preference to Laura Knoff at (510) 486-4750. } E-Mail address = KNOFF-at-LIPOVX.LBL.GOV } Phone = (510) 486-4088, if leaving a message please speak slowly and clearly. } Price: $17 for Loin, $14 for Chicken, $13 for Lasagna,$8 for student } members. } Avoid standing in line by pre-paying. Send your check, payable to NCSM, } to NCSM c/o Laura Knoff } LawrenceBerkeley National Lab } Bldg 1, Room 264 } 1 Cyclotron Rd } Berkeley, CA 94720 Sorry about the format (or lack of it).
TEMSCAN WITH 120KV, PURCHASED 7/82, UNDER SERVICE CONTRACT, ASID W. FREE LENS, SCAN ROTAT. AND TILT, EXT. WAVE FORM MONITOR, MAGNETIC SPEC. POLE PIECE, SEG, 4 HOLDERS INCLUD. OXFORD COLD, LIQ. N2 BAFFLE, GATAN CAMERA (NEGOTIABLE) ASKING $15K, WILL COST $12-15k TO MOVE AND SET-UP. MORE QUESTIONS: GARONEL-at-POLAROID.COM
In message 2 May 96 11:39:25 EDT, "John Grazul" {GRAZUL-at-BIOLOGY.RUTGERS.EDU} writes:
} Freeze Etchers, } } Besides Tech no Trade (who has yet to send me a new phone number, } catalog, address) where might I get platinum pellets, hollow } carbon, and filaments for my BAF 301? I have talked to a company } that will help me out with the carbon and platinum, but the tungston } filaments are another story, or maybe techno trade can get me their } number so I can throw a couple of hundred bucks their way. } } } } John Grazul } Rutgers University } Electron Microscope Facility } ---------- Try contacting Albrecht (Albi) Auwater or Johnny Hagen at:
TECNO TRADE INTERNATIONAL 7 perimeter Road Manchester, NH 03103-3343 Tel (603) 622-5011 Fax (603) 622-5211
************************************************************************* M.V. Parthasarathy Professor of Plant Biology & Director, Cornell Integrated Microscopy Center 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 am writing for a colleague of mine who is interested in puchasing an updated ultramicrotome. Does anyone have a list of companies that sell microtomes? The two I know are RMC and Leica. Are there more? Also, does anyone have a preference for one microtome over another?
Please send responses to Terry Colberg at colberg-at-okway.okstate.edu.
Thank you in advance,
Ginger R. Baker EM Lab Manager Department of Anatomy, Pathology, and Pharmacology 250 Veterinary Medicine Oklahoma State University Stillwater, OK 74078 (405) 744-6765 FAX: (405) 744-5275 Email: lizard-at-okway.okstate.edu
} Freeze Etchers, } } Besides Tech no Trade (who has yet to send me a new phone number, } catalog, address) where might I get platinum pellets, hollow } carbon, and filaments for my BAF 301? I have talked to a company } that will help me out with the carbon and platinum, but the tungston } filaments are another story, or maybe techno trade can get me their } number so I can throw a couple of hundred bucks their way. } } } } John Grazul } Rutgers University } Electron Microscope Facility John, I just got my order from them. It came quite quickly, actually. The address from the to of the invoice reads: Technotrade International 7 Perimeter Road Manchester, NH 03103-3343 Tel (603)622-5011 Fax (603)622-5211
As far as part numbers, I think they use the old Balzers catalog item numbers. The tungsten cathodes I ordered were item number BU020 23T. Sorry, I didn't order any Pt pellets or hollow carbon. The Balzers numbers for these items are: Pt inserts, 1.5X2mm #BD481 505, hollow carbon=#BD484 055. Give them a call and see if they cross-reference. I threw a couple hundred bucks their way.
} MY QUESTIONS ARE: } 1. Does anyone out there have similar problems with this configuration of } TEM? } 2. Is this a problem with most TEMs (small specimen chamber etc)? } 3. Has anyone any suggestions about how to minimise or remove this problem? }
Malcolm,
I noticed your posting last week concerning spectral artefacts from your Hitachi H7000 TEM and H7110 STEM (STEM & SEM); Link QX2000 EDS x-ray analysis system. Since I have not seen any replies on the Listserver I thought it was time to pass on my own experiences.
Our microscopes are JEOL 2000 fx and 2000 fxII TEM's with high take-off angle detectors (~72=B0). The detectors are old Tracor Northern units with new Link ISIS microanalysers. Both are almost exclusively operated at 200KV in TEM mode. We frequently observe Fe-K and compton scattered Mo-K lines in spectra. The Iron is presumable fluoresced in the specimen area by the compton scattered Mo-k x-rays which are generated in the condenser system.
Replacing the selectable and/or fixed condenser apertures by very thick Mo or Pt apertures should eliminate the compton scattered Mo-k x-rays (or so I'm told, I'm still trying to get part numbers for my microscopes). Removing the condenser aperture strip is not a good idea since the electrons will be sprayed over a much larger area of the sample/specimen chamber than just the area you are analysing (hence the larger Fe peak).
If you try the thick aperture as I've suggested, or any other fixes, please let us know how successful it is. Regards,
Mark Blackford TEM Group Materials Division, Ansto PMB 1, Menai, N.S.W. Australia 2234 Phone (02) 717 3027 =46ax (02) 543 7179
} I have been using a Hitachi H7000 transmission microscope for several years } and my x-ray spectra are often affected by background Mo peaks and } occasionally Fe. Fe is presumably coming from scattered electrons hitting the pole-pieces or something else in the region of/below the specimen, but this will depend on geometry and I do not know the Hitachi.
Is the Mo peak really Mo? Check that it is not slightly lower in energy i.e. about 17.38keV rather than 17.44keV. If it is then the problem is due to Compton scattering, probably in the various grahite inserts, of Mo X-rays. The Mo X-rays will be from thin Mo condensor apertures. This is elliminated by the use of thick apertures. (See reference AR Wilson and LT Lambrianidis, J. of Microscopy, vol160, pt1 Oct 1990, pps 1-7.)
} The objective aperture seems to be the biggest contributor of Mo and when I } removed the aperture strip from the microscope the Mo disappeared but I got } more Fe (presumably from the holder tip). This would be inconvenient anyway } because of the machine down time etc.
What is the objective aperture made out of? If Mo then the problem is due to scattered electrons hitting the aperture (that's what it is for).
alan.wilson-at-dsto.defence.gov.au Dr Alan Wilson Senior Research Scientist Ship Structures and Materials Division Aeronautical and Maritime Research Laboratory Defence Science and Technology Organization 506 Lorimer St Fishermens Bend 3207 Victoria Australia ph 61 3 9626 7508, fax 61 3 9626 7816 or 61 3 9626 7087
Our lab is in need of a Cryo-Nova system as a backup and spare parts source. If anyone has one in storage or permanantly off-line and is willing to part with it for a minimal amount of money, please contact me.
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Customer Support Engineer ************************* Due to continued growth in its biomedical division, Applied Precision, Inc. is presently seeking a Customer Support Engineer to work for the DeltaVision product line. DeltaVision is an optical sectioning, deconvolution microscope for high resolution, fluorescence imaging in 5-dimensions.
We are seeking a person with a technical background, experience in optical microscopy and research instrumentation, and experience with or aptitude for using SGI/Unix-based computer systems. The person must have excellent communication skills. Responsibilities include customer support, system installation and service, and support of conferences and workshops. Extensive travel is required.
Ideal candidates are interested in learning about and helping others to understand and use new technology. You must be comfortable working in a team-based environment, and be able to work independently.
This is a very good opportunity to contribute to a rapidly growing product that makes a significant contribution to the field of microscopy.
For more information about this opportunity, send your cover letter and resume to:
Applied Precision, Inc. 8505 SE 68th St. Mercer Island, WA 98005 Attn: Pete Williams
email: info-at-api.com
Resumes will be held in confidence.
________________________________________________________________________ Dean Waters DeltaVision Systems Applied Precision, Inc. 8505 SE 68th Street Mercer Island WA 98040
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Alas, I typed my home zip code rather than our company zip code in the address noted below. Please forgive the double posting, and note the correction.
Customer Support Engineer ************************* Due to continued growth in its biomedical division, Applied Precision, Inc. is presently seeking a Customer Support Engineer to work for the DeltaVision product line. DeltaVision is an optical sectioning, deconvolution microscope for high resolution, fluorescence imaging in 5-dimensions.
We are seeking a person with a technical background, experience in optical microscopy and research instrumentation, and experience with or aptitude for using SGI/Unix-based computer systems. The person must have excellent communication skills. Responsibilities include customer support, system installation and service, and support of conferences and workshops. Extensive travel is required.
Ideal candidates are interested in learning about and helping others to understand and use new technology. You must be comfortable working in a team-based environment, and be able to work independently.
This is a very good opportunity to contribute to a rapidly growing product that makes a significant contribution to the field of microscopy.
For more information about this opportunity, send your cover letter and resume to:
Applied Precision, Inc. 8505 SE 68th St. Mercer Island, WA 98040 Attn: Pete Williams
email: info-at-api.com
Resumes will be held in confidence.
________________________________________________________________________ Dean Waters DeltaVision Systems Applied Precision, Inc. 8505 SE 68th Street Mercer Island WA 98040
} I have been using a Hitachi H7000 transmission microscope for several years } and my x-ray spectra are often affected by background Mo peaks and } occasionally Fe.
Dear Malcolm, I have a Hitachi H-800 200 kV STEM with an Ortec EDX system at high take-off angle: 68 degrees. Unlike you, I cannot run the EDX with the objective aperture in place, since the back scattered electrons and X-rays from the Mo objective aperture flood and "kill" the EDX system. However, I can run the EDX system with the sample untilted. Generally, I find the spectrum is very clean and free of any artifacts, but if I count for a long time I get a small Fe and Co peak, presumably from the inside of the sample chamber. Can you remove the Mo objective aperture, or swing it out of the way? Apparently, the main column modification required for optimum EDX operation in a STEM is shielding on the moveable condenser aperture. My system has a pure graphite fixed condenser aperture and a thick Mo moveable aperture with lead shielding above and graphite covered aluminum below. This is the modification required for an effective EDX STEM system. Do you know if this system is in your TEM? The only other help I know of is to use carbon-coated nylon grids or BE grids to reduce scattering from the grid bars. The only other solution I have heard of is to cover the objective pole-piece with a shaped insert of beryllium or possibly painting it with carbon DAG. The secret is to figure out where the electron and scattered X-rays are entering your detector or alternately what your detector is "looking" at, and shield it in some way. I hope this is some help. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
} My problem is that this microscope appears to exhibit something } similar to astigmatism even at the lowest magnification. We also have
} Please could you send any thoughts and suggestions to me at } DAVIES-at-ASUKA.NAL.GO.JP } } Any ideas would be gratefully received. Thank you. } } Dr. Ian Davies
Dear Ian,
If the plane of the object being viewed is not at 90 degrees to the axis of the microscope, it can offer a view similar that of an astigmatic system.
I purchased an RMC MT7000 almost two years ago and have been very satisfied with its performance. I think we needed service once. I have been impressed with their technical and service support.
No commercial endorsement should be implied.
-- David Rothbard Institute of Paper Science and Technology
We have had this problem several times. The diaphragm in the outlet is probably clogged with some foreign substance. This diaphragm is found inside a nut, holding the outlet pipe on rear side of the CPD030. It is easily removed. Inspect the small hole of the diaphragm in a stero microscope.
It has happened, especially on the earlier instruments in the series, that small Teflon pieces from the valves obstruct the outlet. This can happen because the valves are placed after the filter.
Good luck!
Rolf Odselius
} We have a Bal-Tec CPD030. It has been used very little. This is } because we cannot get it to drain. It might take all night for the } alcohol-CO2 mixture to empty. The instructions seem very } straigt-forward. We fill partially with 100% ethanol (with or } without a sample), cool to 8 degrees C, let in the carbon dioxide } (yes, the tank is the kind with the dip tube), then press the } medium out button. At first it seems to empty, then gets very slow. } Has anyone had a similar problem? The valves seem to work as I } hear a loud click when pressing buttons. The filters are clean. } Any other ideas? thanks. _________________________________________________________________ Rolf Odselius, PhD |E-mail: Rolf.Odselius-at-emu.lu.se Electron Microscopy Unit |Phone: +46 46 171075 office University Hospital | +46 46 293692 home S-221 85 Lund, Sweden |Pager: +46 740 288992 Minicall http://www.emu.lu.se/ |Fax: +46 46 172975 http://www.ldc.lu.se/~scandem/ |Cellular: +46 70 5581085 GSM/SMS
In message {v02130504adb3b23e3508-at-[199.77.235.102]} David Rothbard writes: } I purchased an RMC MT7000 almost two years ago and have been very satisfied } with its performance. I think we needed service once. I have been impressed } with their technical and service support.
We bought a new MT-7000 about 4 years ago. Over all its been a good performing ultramicrotome for us in a multiple user lab with many users. Initially we had a problem with the fine mechanical advance slipping on the knife stage. Eventually the stage was replaced under warranty with a new one. Some accessory parts on the original order, such as water trough filling device, blockface viewing mirror, service manual, were slow in coming(1-2 years), but finally arrived and it is good solid equipment. (As an aside, thats not the first time I've bought "ghostware": "Yes it has a part number and yes its described in the glossy brochure and yes you paid for it and yes.....we're working on it".)
We like the wide range of controls and flexibility and ease with which it can be operated and with which new sectioners can learn to use it.
This view is not a commercial endorsement, just the experience and opinion of one ultramicrotome supervisor and user.
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"MICROSCOPY & MICROANALYSIS 96" in Minneapolis, Minnesota, Aug.11-15, 1996
The Department of Physics, University of Alberta, Edmonton, Canada is advertising a tenure-track Assistant Professorship in Experimental Condensed Matter Physics and Materials Science, starting 1 Jan 1997.
Preferred areas of research are: scanning-probe microscopy, nanostructures, thin films, surface physics and high-temperature superconductors.
For further information, please contact egerton-at-phys.ualberta.ca or dtlunty-at-phys.ualberta.ca
At 12:56 PM 4/25/96, Pat Kingman wrote: } } Our lab would like to convert our Zeiss Axiomat from film to digital. } We have no direct experience, and would appreciate any comments or } experiences that would help us research the best route to take, both } hardware and software, but particularly the former. } } I believe that this topic has been discussed recently, but unfortunately } I was not concerned until just now! If anyone has saved either their own } previous postings or those of others, I would appreciate if you would } email to me directly if you consider it repetitious to repost. }
Perhaps some info about my experiences would help.
I am a microscopist in an industrial research lab. We have electron and optical microscopes in many varieties. Our optical microscopes are made by Zeiss, Leitz, Wild, B&L and Nikon and include stereos, low-mag macros, and conventional research-grade microscopes. I set up analog video imaging on our optical optical microscopes seven years ago.
Seeing a need to keep copies of images, I switched from analog video imaging to digital imaging on our optical microscopes last year. A "standard" microscope workstation consisted of the following:
1. Microscope 2. Optical coupler for video camera 3. Video camera 4. Computer 5. Image capture board 6. Image storage device 7. Printer
The Optical couplers were from Diagnostic Instruments, ~$300.
The video camera was the Hitachi CV-20, a 3-chip camera with C-mount. It has great image quality and light sensitivity! It cost around $4500.
The computer was a 486 PC. I would recommend a Pentium, the fastest you can afford. The 486 works OK, but is a little slower.
The image capture board was a TARGA+ from Truevision. This is an older board, but it was matched to our Kodak 450GL printers. I would recommend a newer board so as to be more compatible with currently available software. Boards run ~$2000.
The image storage device was to be a recordable CD-ROM, however because of the large number of images we expected to generate and the need to have computer network access to those images, we installed an optical disk "jukebox" capable of storing 40gB of images from both the optical and electron microscopes. All microscope workstations are on a network which accesses this "jukebox". Recordable CD-ROMs run ~$1,000.
The printer was a Kodak 450GL dye-sub printer. Kodak has since discontinued the manufacture of these printers. A real shame because they are really quite good. Also, it met our need for a ~4X5 inch print. Sony and Hitachi make similar printers. I don't know about their pricing. When we need higher quality images on an 8 1/2 X 11 inch format we use a Kodak 8600PS dye-sublimation printer. We use it for both electron and optical micrographs. It cost ~$9000.
Everyone who uses this system is quite happy with the ease of use and quality of image output. The only thing we lack at this point is a good database system for our images. There are several available, but we have not found one that meets all of our specialized needs. As a last resort, we plan to develop our own if we haven't found one in a few months.
I hope this helps you and anyone wanting to get into digital imaging.
Dr. Dennis B. Barr Eastman Chemical Company Microscopy and Morphology Research Laboratory P.O. Box 1972 Kingsport, TN 37662-5150
We have a JEOL 2010 TEM and need to measure the beam current either just prior to or just after EDXS analysis, without the desaturation of the filament (LaB6) or removal of the sample holder. We currently use a Gatan cryo/analytical holder which has a Faraday cup in on it but the position of the cup is beyond the limit of travel of the goniometer. We also have a normal (non-analytical) JEOL sample holder.
The TEM doesn't have a backscatter detector so we can't use that method. We do have a Gatan imaging filter/EELS system and could measure the beam current on the drift tube of this but the setup for EDXS is different from that of EELS and also we would prefer to measure the current at the sample!
Does anyone have any ideas as to how we could measure the beam current at the sample with our current equipment as we don't want to have to purchase yet another holder?
I have digital micrographs of latex spheres and need to calculate a radial intensity profile (grayvalues vs. radius). Does anybody know if for this a macro for NIH Image is available?
Hej, titta p=E5 detta! Snyggt. Och det b=E4sta av allt, portotarifferna finns ocks=E5!
Rolf
------- Forwarded Message Follows -------
Grattis!
Du dr anmdld som smygtittare och har gjort dig fvrtjdnt av adressen http://www.torget.se I morgon blir denna adress officiell, du fer adressen redan idag!
Vdlkommen till Torget,
Redaktionen
P.S. Passa pe att anmdla dig som medlem pe Torget redan nu! D.S.
_________________________________________________________________ Rolf Odselius, PhD |E-mail: Rolf.Odselius-at-emu.lu.se Electron Microscopy Unit |Phone: +46 46 171075 office University Hospital | +46 46 293692 home S-221 85 Lund, Sweden |Pager: +46 740 288992 Minicall http://www.emu.lu.se/ |Fax: +46 46 172975 http://www.ldc.lu.se/~scandem/ |Cellular: +46 70 5581085 GSM/SMS
} I have digital micrographs of latex spheres and need to calculate a } radial intensity profile (grayvalues vs. radius). Does anybody know } if for this a macro for NIH Image is available?
This is a special case of a useful calculation sometimes referred to as "azimuthal averaging". It is also useful in "reciprocal space", for example in improving signal to noise in powder diffraction profiles, or when locating image contrast transfer function zeros. We use a verb in Semper6 for this purpose, which also offers a way to locate the center of a circle (or your sphere) given 3 points on its perimeter. For diffraction patterns, we've put together a Semper macro for statistically refining the center determination with more than one triplet of points on one or more "rings" around the center, and calculating an uncertainty in that determination to boot.
If there is interest in the macro let me know. I am not sure where to find out about the availability of the Semper6 language itself these days, however.
Within NIH Image, there is a macro file called "Plotting macros". Load this file into NIH Image under the special menu, and use the one called "Radial Intensity Distribution..." It's just that. You pick the point on your image from which to do a plot (use the select lines tool to choose the point), the number of lines to do profiles along around the point, and finally the radius of the lines. It will plot a composite histogram of all the profiles.
A colleague of mine is setting up her own microscopy prep lab and has asked where we purchased our angular and linear film measuring device. Ours comes from the Charles Supper Co. which doesn't seem to exist any more. The microscopy supply catalogues do not have a similar device. What we want is a device that would sit on top of a light table. We don't want to use a lupe to measure angles. Does anyone know of a supplier for such a tool. Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets The Johns Hopkins University Baltimore, Maryland 21218
Does anyone have an address for a USA distributor of the Sn/Pd colloidal coating solution made by:
Neyco s.a. Paris, France
Thanks! Phil
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
Further to the recent discussion on "good" buffers, has anyone besides me had problems with osmium made up in PIPES?
I tried using PIPES buffer many years ago after reading the 1973 paper of Salema & Brandao, and got very nice results (with fungal tissue at that time) but the osmium solution went brown within half an hour of being made up in PIPES (diluting 2 percent Os in water to 1 percent with double-strength PIPES). I made up the buffer several times from the same batch of PIPES powder but always got the same result. So I went back to cacodylate.
Was it just me? Did I perhaps have a "bad" batch of "good" buffer?
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department School of Medicine University of Auckland Private Bag 92019 Auckland New Zealand
Message-Id: {199605080150.VAA16092-at-mime2.prodigy.com} X-Mailer: Prodigy Internet GW(v0.9beta) - ae02dm02sc06
-- [ From: Charles A. Garber., Ph.d * EMC.Ver #2.10P ] --
The following was written: =============================== Does anyone have an address for a USA distributor of the Sn/Pd colloidal coating solution made by:
Neyco s.a. Paris, France
Thanks! Phil ========================================= SPI Supplies has been the representative for NEYCO S.A. for North America. We can supply you with the product.
The technique was developed by French researchers and NEYCO has the world wide rights (as I understand it) to distribute this product. You can contact NEYCO directly as follows: {neyco-at-imaginet.fr} .
For your further information, NEYCO has been a long time distributor in France of products for materials science research. They are the French distributors, among other firms, of products of SPI Supplies and also South Bay Technology. The person to contact is Ms. Isabel Richardt. They are located in a Paris.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: spi2spi-at-2spi.com
cc ___________________________________________________________
Stefan Andreatta Institute of Zoology and Limnology, University of Innsbruck Technikerstrasse 25, 6020 Innsbruck, Austria phone: fax: {Stefan.Andreatta-at-uibk.ac.at}
Damian, Try Microlites Scientific, ph. 1-800-263-8902. Talk to Frank Nasser. They are located in Canada, but I had no trouble getting bulbs sent to me in Dallas, TX. I went to them when my local sources didn't have what I needed and they were very helpful. Good Luck.
Karen
On Tue, 7 May 1996, Neuberger, Damian wrote:
} } Hi All: } } Can anyone recommend a source of microscope illuminator bulbs? I'm looking } for OSRAM 64625, 12V/100W and Olympus 6-8V5ATB-1, 6V/30W, a DDL 150W 20V } bulb for our fiber optic light source and similar types of bulbs. Thanks } for any suggestions you may have. } } Damian Neuberger } neuberd-at-baxter.com }
I used osmium in HEPES buffer for years although I have recently switched to using unbuffered osmium in water with no noticeable difference.
} Hi everyone, } } Further to the recent discussion on "good" buffers, has anyone } besides me had problems with osmium made up in PIPES? } } I tried using PIPES buffer many years ago after reading the 1973 } paper of Salema & Brandao, and got very nice results (with fungal } tissue at that time) but the osmium solution went brown within half } an hour of being made up in PIPES (diluting 2 percent Os in water } to 1 percent with double-strength PIPES). I made up the buffer } several times from the same batch of PIPES powder but always got the } same result. So I went back to cacodylate. } } Was it just me? Did I perhaps have a "bad" batch of "good" buffer? } } } } Regards } } Stephen Edgar } } Electron Microscope Unit, Pathology Department } School of Medicine } University of Auckland } Private Bag 92019 } Auckland } New Zealand } } email address: s.edgar-at-auckland.ac.nz } Phone : +64-9-3737599 extn 6473 (GMT + 12h) } Fax : +64-9-3737459
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)
} } SHORT COURSE ANNOUNCEMENT } } ------------------------------------------------- } } FUNDAMENTALS AND APPLICATIONS OF LIGHT MICROSCOPY } } ------------------------------------------------- } } JUNE 16-21, 1996, Burlington Vermont } } } } www.microscopyed.com
Experienced microscopy problem solvers will teach a 5-day hands-on course on achieving the maximum information from light microscopy. The emphasis of the course will be to provide hands-on experience, and the background for interpretation of images. } } The course will cover the principals of light microscopy, contrast techniques for the microscope, adjustments of the microscope for optimum contrast and resolution, interpretation of images in terms of light-matter interactions and image recording. } } A full range of reflected and transmitted light microscopes, as well as contrast equipment, will be provided for use by the students. Students are encouraged to bring their own samples. } } This course will be applicable to all disciplines using light microscopy. Although ideal for beginners, it is designed as an advanced workshop, with the opportunity to concentrate on light microscopy in a relaxed environment. ____________________________________________________________ } } Faculty: Philip C. Robinson, ret. Staffordshire University, UK, Dept of Ceramics, author of the RMS Microscopy Series book, Applied Polarized Light Microscopy. Robert Hoffman, inventor of the Hoffman Modulation Contrast system, Mary. McCann, course organizer, Robert Janes, Metropolitan (London) Forensic Science Laboratory, and Dennis O'Leary, MicroOptical Methods. Vermont Optecs, research instrument specialists, will supply a variety of microscope equipment for the course. } } For further information:www.microscopyed.com } } For course brochure and registration, contact Mary McCann, course organizer, e-mail: mccanns-at-tiac.net telephone 617-484-7865 fax: 617-484-2490
Over the years I've tried the Good buffers for various (mostly botanical) specimens. OsO4 will react with (at least) the piperazine containing ones, likely in competition with what you're trying to fix in the first place. I generally use OsO4 in water, unbuffered, with no apparent drawbacks.
A second consideration is that the buffers that Good's group were working on were for physiological experiments, and one of their criteria was that the buffer should not cross biological membranes. I'm not sure this is good (so to speak) for ultrastructural preservation, if there is an appreciable lag between introduction of the first fixation cocktail and the loss of membrane selectivity.
For a good basic discussion of buffers, in general, I've always liked D.E.Gueffroy's booklet called (not supprisingly) Buffers, from Calbiochem Biochemicals, Behring Diagnostics division of Americanm Hoechst. Also for Good buffers there is Good, et al. 1966. Biochem 5:467.
I use cacodylate buffers mostly now, too.
Cheers, John Heckman
TEM supervisor Center for Electron Optics Michigan State University .} } Hi everyone, } } Further to the recent discussion on "good" buffers, has anyone } besides me had problems with osmium made up in PIPES? } } I tried using PIPES buffer many years ago after reading the 1973 } paper of Salema & Brandao, and got very nice results (with fungal } tissue at that time) but the osmium solution went brown within half } an hour of being made up in PIPES (diluting 2 percent Os in water } to 1 percent with double-strength PIPES). I made up the buffer } several times from the same batch of PIPES powder but always got the } same result. So I went back to cacodylate. } } Was it just me? Did I perhaps have a "bad" batch of "good" buffer? } } } } Regards } } Stephen Edgar } } Electron Microscope Unit, Pathology Department } School of Medicine } University of Auckland } Private Bag 92019 } Auckland } New Zealand } } email address: s.edgar-at-auckland.ac.nz } Phone : +64-9-3737599 extn 6473 (GMT + 12h) } Fax : +64-9-3737459 }
Via: uk.ac.bbsrc; Wed, 8 May 1996 17:31:37 +0100 X400-Received: by /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Wed, 8 May 1996 17:33:16 +0000 X400-Received: by mta arcs.bbsrc.ac.uk in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Wed, 8 May 1996 17:33:16 +0000 X400-Received: by mta LARS.MUAS in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Wed, 8 May 1996 17:32:18 +0000 X400-Received: by mta LARS in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Wed, 8 May 1996 17:32:20 +0000 X400-Received: by /PRMD=UK.AC/ADMD= /C=GB/; converted (ia5-text); Relayed; Wed, 8 May 1996 17:32:29 +0000 Receipt Notification Requested) (IPM Return Requested)
Dear Fellow microscopists,
Can anyone help me solve a problem I have with UV-cured butyl-methyl-methacrylate? I have been using this resin for many months, deresining with acetone and immunolocalizing tubulin and F-actin in secondary vascular tissues of horse chestnut. Great, the system works well, apart from one small niggle: I have been unable to prevent formation of bubbles/vortices within the the resin as it polymerizes. Usually this is not a problem because the tissue blocks are dense enough to stay on the bottom of the cylindrical embedding capsules that I use. However, it is a problem with young tissue and root tips - which are not as dense - and often the tissue floats up to the surface of the resin on the bubbles and is generally unusable. Does anyone have a cure for this phenomenon? I have tried placing the resin-filled capsules under vacuum for 30 min prior to curing but not avoided bubble formation... Any suggestions will be most welcome. Thank you,
Nigel Chaffey
------------------------------------------------------------------------------- Internet: nigel.chaffey-at-bbsrc.ac.uk IACR-LARS, Dept. of Agricultural X400:G=nigel; S=chaffey; O=bbsrc; P=uk; C=GB Sciences, University of Bristol, Tel: +44 (0)1275-392181 ext:230 Long Ashton Research Station, Fax: +44 (0)1275-394281 Long Ashton, Bristol, BS18 9AF -------------------------------------------------------------------------------
An internal message within our department has of a presently unknown technical reason been sent to the Microscopy List. A posting I did send to the list did not turn up though. We must have had some kind of cross-over in our mailserver. Sorry folks!
Maybee e-mail isn=B4t as safe as it is said to be... And thanks for all kind remarks about our "exotic" language :-)
Rolf _________________________________________________________________ Rolf Odselius, PhD |E-mail: Rolf.Odselius-at-emu.lu.se Electron Microscopy Unit |Phone: +46 46 171075 office University Hospital | +46 46 293692 home S-221 85 Lund, Sweden |Pager: +46 740 288992 Minicall http://www.emu.lu.se/ |Fax: +46 46 172975 http://www.ldc.lu.se/~scandem/ |Cellular: +46 70 5581085 GSM/SMS
Bob, I've been examining tissues of all types for a good many years and there are usually several options (preperation wise) that are dependent on what you want to see.
Need a couple more specifics ie: type of tissue and what you want to look at. Given these I may well be able to poit you in the right direction . regards, bob
Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis University of North Carolina CB#7400, Rosenau Hall Chapel Hill, NC 27599-7400 Ph. 919-966-6343 Fax 919-966-6123
A friend is interested in purchasing a film recorder (slide maker) for producing high resolution color slides or transparencies. This is a professional facility where hi res is important and price is a side issue. Please send suggestions and comments to: jrichard-at-siu.edu
Thank you.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
Dr. David G. Gantt Phone: 1-912-681-5964 Dept. of Biology Fax: 1-912-681-0845 Landrum Box 8042 e-mail: david_gantt-at-gsvms2.cc.gasou.edu Georgia Southern University Statesboro, Georgia 30460-8042
The problem you describe sounds very similar to what is often encountered with methylmethacrylate embedding. The bubbles usually form right at the end of the polymerization reaction when the medium is so viscous that they cannot escape to the surface.
Most often the bubbles are from boiling monomer which nucleate on your tissue. You need to address ways of lowering the temperature. I would suggest: - decreasing the volume of monomer polymerized - lowering the concentration of initiator or catalyst - placing the molds in a volume of water to act as a heat sink - performing the polymerization under pressure (raising the boiling pt)
Obviously not all of these are practical solutions. Still, you should find a way of lowering the maximum temperature that the solution achieves.
Good luck, Joe Tabeling Delaware Diamond Knives 3825 Lancaster Pike Wilmington, DE 19805 800-222-5143 FAX:302-999-8320 http://www.ddk.com
I use LM with polarized light and contrast liquids (potassium iodo mercurate in water/glycerol) at 200 and 500 X. For Chrysotile I use also nitrobenzene or mixtures of cinnammic aldehyde and benzyl alcohol. I have a Chrysotile standard. My problem is to manage other asbestos fibers, if present. How I can see the other asbestos fibers: Crocidolite, Tremolite, Anthophyllite, Amosite and Actinolite ? I don't have standards with those fibers. Some of you can help me ?
I have just obtained my dream microscope and and trying to get set up in my home for a small lab. I need to purchase a microtome. Also, I would like a line on glassware and slides. I have the VWR Scientific catalog but was wondering if there are other vendors I might write to for catalogs.
If any of you have phone numbers or addresses I would appreciate it. Also, if anyone has a used microtome for sale I would love to hear from you.
--------------------------------------------------------------- Eric Chapman - Eagle River Alaska _/_/_/ _/_/_/ _/_/ Hemodialysis Patient _/ _/ _/ _/ _/ _/ _/_/ _/ Sic Gorgiamus Alles _/ _/ _/ _/ _/ Subjectamos Nunc _/_/ _/_/_/ _/_/ ---------------------------------------------------------------
The bubbles are probably being formed during polymerization. I have seen this happen with Epon-Araldite and GMA However we did not use UV polymerization. We polymerized at 4o C in a vacuum dessicator and it eliminated most of the problem. Another trick that I used to use on tissues that had a tendency to float was to place a small amount of resin in the capsule, allow it to to partially polyermrize, place the tissue in the mold and then fill it up with the plastic resinand allow to cure. We also degassed our resin for at least an hour prior to filling the capsules. regards bob
Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis University of North Carolina CB#7400, Rosenau Hall Chapel Hill, NC 27599-7400 Ph. 919-966-6343 Fax 919-966-6123
} While following the thread on pump rebuilds, I wondered if some of you have } extended experience with the ages of roughing pumps? What are the ages } typically achieved with the usual routine maintenance?
We have several belt-drive pumps which are chugging along quite happily after 20+ years, but the direct drive ones have somewhat shorter lives in our experience.
} I recently had two Edwards pumps fail on me. The cause seemed to be } excessive water build-up in the oil chamber that caused rusting. These } pumps are used only to prepump film desiccaters.
We also have found problems here. The best pump for this seems to be a small belt-drive Edwards. We also have a very long path between the darkroom and the pump, which might help with the water and CaSO4. I'm afraid that this service is just very hard on pumps. Yours, Bill Tivol
} Am looking for replacement parts for the ion getter pump on the Zeiss EM109, } in particular, the getter grids. The pump is a Leybold-Heraeus IZ80.
David
We just turned off our EM109 because of the ion getter pump. The grids can be repaired with strips of titanium. For some reason we had a dozen or so in the lab. I was able to make many repairs to the pump before the ceramic insulators started to break down. Zeiss wanted a fortune for the replacement parts. The instrument was not used very much so we are in the process of selling it. A general announcement to the microscopy list will be made in June.
Leybold USA does not sell the replacement parts, but Duniway Stockroom (800-446-8811) was interested in repairing the Triode. However, they could not guarantee that they could fix it.
Good luck
Mike
=========================================================== Michael Dunlap lab (916) 752-0284 Facility For Advanced Instrumentation fax (510) 422-2282 University of California mrdunlap-at-ucdavis.edu Davis CA, 95616 http://128.120.187.6/ ============================================================
Is there anyone in the EM community who has an operating manual or schematics, anything at all, on a Varian VE-10 vacuum evaporator. I have acquired one that seems to be in reasonably good condition and would like to use it for some experiments. However, we have not found any documentation on this particular model evaporator here. It is a VE-10, model 961-1000, SN 1151. Varian have not been able to find anything on this because the model was last made in the 1970's and they sold the model to a company called Thermionics, based at the time in Hayward, CA. My hope is that someone out there has a working model and would be willing to send me a Xerox of the operating manual and any schematics so that we can trouble shoot the one we have here.
Kenneth A. Taylor, Ph.D. Phone: 904-644-3357 Institute of Molecular Biophysics Fax: 904-561-1406 Florida State University E-mail: taylor-at-sb.fsu.edu Tallahassee, FL 32306-3015 Home pages: http://www.sb.fsu.edu/~taylor/ http://www.fsu.edu/~biology/faculty/kat.html
My thanks to all those who responded to my question regarding beam current measurement on a JEOL 2010. It appears that Gatan didn't include the "spacer" to pull the sample holder back that 5mm. I've contacted the agents in Australia and they are looking into it.
Thanks again
Colin Veitch ##################################################################### # # # Colin.Veitch-at-geel.dwt.csiro.au # # Instrumentation Scientist # # CSIRO Division of Wool Technology Tel. +61 (0) 52 275611 # # P.O. Box 21 Fax. +61 (0) 52 275657 # # BELMONT Vic 3216 # # Australia # # # # "We see the Universe the way it is because if it were different, # # we would not be here to observe it." # # # #####################################################################
Message-Id: {9605091859.AA5350-at-pho903.sbphrd.com} To: Microscopy {Microscopy-at-Sparc5.Microscopy.Com}
Position available as advertised in New Scientist (Thursday 2nd May 1996) VACANCY FOR A BIOLOGIST - ANALYTICAL SCIENCES, Harlow, Essex, UK. You will join our microscopy and flow cytometry group which comprises a number of talented scientists who provide the creativity and expertise needed in a large and challenging R&D environment. They have one of the longest-established and best-equipped units of its kind in the industry and exploit state-of-the-art facilities across a broad spectrum of research areas and development activities, including neurosciences, vascular biology and anti-infectives research. You should have a proven interest in any major area of modern microscopy and/or flow cytometry, preferably at doctoral/postdoctoral level, backed by demonstrable skills. This includes any aspect of biological light microscopy and electron microscopy, including cryoTEM-electron crystallography of biomolecules and biological atomic force microscopy. Ref: BAS/NS.
Please do not reply to the sender of this e-mail, but send your CV directly to: The Human Resources Manager, SmithKline Beecham Pharmaceuticals, Old Powder Mills, Leigh, near Tonbridge, Kent TN11 9AN, UK. Closing date: 26 May 1996.
I am trying to find out if other labs have bulk rates for use of their electron microscopes. You may recall that there was a series of messages regarding problems the University of Hawaii had with federal audits. This request is related to that thread. As a part of Dartmouth's effort to comply with the various cost accounting regulations (OMB, A-21, A-133, etc.), I need to try to get an idea of how other labs are handling bulk time purchases from research labs in their institutions. Specifically, do other labs permit bulk purchases of time, or is everyone required to pay on an hourly (or some other) basis? Is you lab a departmental or institutional facility? What level of support do you receive from the university/college (% of operating budget)? Is there a rational basis for your rate structure?
To get things started -- This is an institutional lab. For in-house users we charge $40 per hour for microscope time, and at the moment, charge $3,000 per year for up to two individuals per grant for unlimited access to the microscopes. We are heavily subsidized by the various parts of the college (to the extent of about 80% of our operating budget). We are working on making the rates more rational without eliminating all activity.
I would be happy to provide a list of responses, if any come in.
Note that the regulations mentione above will effect all colleges and universities that receive federal support, so many of you will probably want similar information at some point in time.
Thanks in advance,
Chuck ************* Charles P. Daghlian, Ph.D. Director, Rippel E. M. Facility Dartmouth College, HB 7605 Hanover, NH 03755 phone 603-646-1039
} Date: Thu, 9 May 1996 06:08:23 -0500 (CDT) } From: MASONW {william.mason-at-bbsrc.ac.uk} } To: Multiple recipients of list {nih-image-at-Soils.Umn.EDU} } Subject: Job Position Available } } PRODUCT MANAGER } BIOMEDICAL RESEARCH IMAGING } } Fast growing company developing and selling computer-based products } for biomedical and biotechnology research imaging, using fluorescent } microscopy, seeks a dynamic, outgoing recent graduate or postgraduate } in biological sciences or related discipline. You might have experience in } imaging or photometric applications, or both, using optical probes for } microscopic applications including ion imaging, FISH or other similar } techniques on living or dead cells. } } You will act as our key product specialist and represent the company to both } end users and distributors worldwide. Computer literacy important, and } interest or experience in biomedical sales or marketing will be of } assistance in this important role. This is a challenging opportunity based } in the attractive university town of Cambridge, England, offering exciting } growth prospects, possibility for worldwide travel and an attractive } compensation package for the right individual. } } Apply with CV by post, fax or email to: } } Dr. William Henderson } Managing Director } Life Science Resources } Church Street Barns } Great Shelford } CAMBRIDGE CB2 5EL } United Kingdom } Tel: 44-(0)1223-845836 } Fax: 44-(0)1223-840342 } Email: HENDERSON -at- LSR.CO.UK }
-Kirk _____________________________________________ Kirk Rogers krogers-at-materials.ecn.purdue.edu OR kirk.a.rogers.1-at-purdue.edu Purdue University, School of Materials Science and Engineering, 1289 MSEE building, W. Lafayette, IN 47907-1289 OFFICE: 317-494-8751 FAX: 317-494-1204 http://materials.ecn.purdue.edu/~krogers
-- [ From: Charles A. Garber., Ph.d * EMC.Ver #2.10P ] --
Renzo Garlasco wrote the following: ================ I use LM with polarized light and contrast liquids (potassium iodo mercurate in water/glycerol) at 200 and 500 X. For Chrysotile I use also nitrobenzene or mixtures of cinnammic aldehyde and benzyl alcohol. I have a Chrysotile standard.
My problem is to manage other asbestos fibers, if present. How I can see the other asbestos fibers: Crocidolite, Tremolite, Anthophyllite, Amosite and Actinolite ? I don't have standards with those fibers. Some of you can help me ? ====================================== For calibration and testing SPI has been offering the original UICC (Union Internationale Centre le Cancer) reference specimens, in 0.1 gram vials, for chrysotile, anthophylite, amosite, and crocidolite. Use as "knowns" for SAED, EDS, and XRD. Use as morphological standars for TEM and SEM. This might not sould like a lot of material but for most people it is a lifetime supply. Contact SPI below for pricing and other information.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: spi2spi-at-2spi.com
} Dear Microscopists: } } Could you please tell me how to well prepare for a TEM sample of SiC } fibers in order to observe their cross-sections??? } } Your help is appreciated very much. } } X.G.NING }
Two possibilities at least:
1 - try microtomy. It seems rather unbelievable but it is maybe the only=20 way to have a rather exact idea of the fiber surface. Fiber surface is=20 important because properties of the composite depend on interface=20 fiber-matrix. You may read a letter I wrote a few years ago:
=ABMicrotomy : a convenient method for preparing Transmission Electron=20 Microscopy in ceramic science=BB, Jour. Mater. Sci. Lett., 9 (1990), 48.
(with a typographic mistake on top of page 49: the summit of the pyramid=20 is .2 mm, not 2mm.) I can send a copy if needed. Please contact me off line.
2 - "Michael K. Cinibulk" {cinibumk-at-ml.wpafb.af.mil} posted a quite=20 interesting message a few months ago, about an improved ion mill method.=20 copy hereafter.
sincerely,
Yves MANIETTE
******BEGINNING OF MESSAGE*****
} Dear Microscopists: } } Could you please tell me how to well prepare for a TEM sample of SiC } fibers in order to observe their cross-sections??? } } Your help is appreciated very much. } } X.G.NING
I recently asked a similar question and received a whole bunch of replies which are reproduced below.
Hope this helps
____________________________________________________________________________ My original message:
Dear all, Has anyone out there prepared non-conducting ceramic fibres for TEM, either across or along axis? If so, how did you do it? We have some mullite fibres we would like to look at and although we have some ideas about how to prepare specimens we would appreciate any ideas or feedback on this topic. ____________________________________________________________________________ We have been routinely preparing ceramics fiber tows (Nextel, Nicalon, HPZ, etc.) with and without ceramic coatings for TEM characterization for about two years now. After much trial and error with a number of methods (some in the literature) we have settled on a rather efficient method of producing high-quality sections. We are just finishing a short communication on this technique, which I could send to you when completed.
First, the tows are vacuum-impregnated with a high-temperature epoxy, such as G-1. It is important that the fiber-volume fraction be high, which can be achieved by either pulling many tows through the tip of a pipette, or alternatively using shrink tubing. The epoxy is then allowed to cure.
The key to obtaining a good thin section with a large electron transparent area is to minimize ion-milling time, which requires mechanically thinning the sample to thicknesses less than 5 um, ideally less than 1 um. We use a tripod polisher with 3M diamond lapping films to minimize differential polishing and surface relief. The epoxy-impregnated tows have been thinned both axially and transversely; either way wil produce good specimens. Again, you want to thin the specimen as much as possible prior to ion milling; if the specimen is too thick the epoxy will be milled away before the fibers and any coating becomes electron transparent, with the fibers usually falling out. We also need the epoxy to bound the coating for coating thickness determination.
The specimen is then mounted on a Cu washer, and maybe a grid, and ion milled at low angles for 30-60 min.
For more details please contact me off line.
Dr. Michael Cinibulk cinibumk-at-ml.wpafb.af.mil Wright Laboratory/MLLM Wright-Patterson Air Force Base, Ohio ____________________________________________________________________________ Do you have a tripod polisher in your prep lab.
We have a way of prepping fibers using a tripod that worked nicely for cross and longitudinal sections of SiC fibers.
Ron Anderson ____________________________________________________________________________ I assume that you are aware of the difficulties in doing this. I know of two methods which work very well and are based on other, relatively well-known techniques for producing cross-sections.
A) embed in resin and ultramicrotome cross-sections; this is only convenient if you have someone who does ultramicrotomy already, in which case it is probably the simplest.
B) sandwich with epoxy between two silicon wafers and then cross-section 'normally,' just like a VLSI device; by visually monitoring optical transmission through the silicon, you can thin the composite structure well below 10 micrometers before ion-milling (which in turn helps mitigate differential milling effects). This works for almost everything. The same basic technique could be used to thin the fiber along the axis.
Please let me know if something easier comes along, since I have to be doing this as well...
Daniel L. Callahan Assistant Professor of Materials Science Rice University http://www.owlnet.rice.edu:80/~dlc/ ____________________________________________________________________________ I had pretty good luck preparing ceramic fibers (mullite, SiC, Al2O3/ZrO, and others), of 10 to 30 micron diameter, for TEM analysis. The method is very simple, and consists of first making a composite out of the fibers and an epoxy, then thining down as with a bulk sample (polishing, dimpling, ion milling). Things to keep in mind:
1. If the fibers are short and tangled as a small cotton ball, saturate the epoxy with the fibers mixing very well. The fibers will brake but that doesn't matter since is unlikely that you will be interested in crystalline defects. Squeeze the composite between two teflon or other non-sticking sheets using glass slides for backings.
If the fibers are long (more than 3mm) you can lay down a thin layer of epoxy on a teflon sheet, and the fibers on top, one at a time or in bundles, in a single direction to cover a strip 3mm wide x length of the fibers. Then squeeze between two glass slides and another teflon sheet covered with epoxy. Cure the epoxy as per manufacturer instructions. In both cases you will be able to form a very thin composite that requires a little grinding and polishing on both sides before punching out 3mm discs, which can be further polish to 100 micron. Dimple as usual (with diamond paste)
2. Epoxy: try with whatever you have available. Some people prefer conductive epoxies. Initially, I used several conductive and non conductive products from TRA-CON (Medford, MA, 508-391-5550), but for some time now I have settled on G1 epoxy from GATAN (412-776-5260).
3. Ion Milling: the mullite I worked with milled very fast probably because it was very porous. Other ceramics mill very slowly compared to the epoxy. This is a problem that can be alleviated using a LN2 stage in the ion mill, to slow down the milling rate of the epoxy, and having as much fiber content as possible in the mixture.
4. Even if you use a conductive epoxy you may have to carbon coat the thinned samples.
Good Luck!
Augusto Morrone Univ. of Florida Materials Science and Engineering P.O.Box 116400 (352) 392-6985 amorr-at-mse.ufl.edu ____________________________________________________________________________ been using the Tripod polisher and a unique technique for tightly binding fibers together. They have been working with SiC fibers, but I think that it would work for you. Their Email addresses are CINIBUMK-at-ml.wpafb.af.mil and SCHELTFJ-at-ml.wpafb.af.mil
- -Scott Walck ____________________________________________________________________________ I think that someone here used a Gatan PIMS to do this several years ago with some success. The length of the fiber went across the 3mm washer. John Hunt ____________________________________________________________________________ Over the years I've prepared various fibre materials in various ways. These all should give results; the quality varies, but then so does the effort required.
1) If the fibres are thin to begin with, pop them onto a support film covered grid, carbon coat, and voila!
2) If you have experience with ultramicrotomy, embed and section small areas.
You may require a coupling agent to promote resin adhesion, and the result of sectioning will be a mass of shattered fragments (unless the fibres are amorphous), but then you might also get a huge amount of viewable area free of chemical contamination.
3) Mix the fibres into a particularly hard resin such as Petropoxy which you can then prepare (cut, grind, dimple, ion mill or tripod) as a monolithic lump. I'm not aware of any resin hard enough to thin at the same rate as a ceramic, but I've prepared SiC fibres this way with success.
4) Create a composite by electroless deposition of metal (usually Ni) which you can then prepare in the ion mill. This one gives by far the best result in my experience, but is also the trickiest and most potentially time consumming. A short electroless plate to make the fibres conductive , followed by electrolytic deposition in standard plating baths is a good option if you have access to such. The method I've used was adapted from metal powder prep. techniques.
Let me know if you'd like further information on any of this.
############################################################### # # # Don Steele STEELE-at-KRDC.INT.ALCAN.CA # # ALCAN INTERNATIONAL # # Kingston Research and Development Center # # P. O. Box 8400 # # Kingston, Ontario Canada K7L 5L9 # # # ############################################################### ____________________________________________________________________________ The answer is simple (at least in principle!) - embed them in a hard epoxy resin or acrylic, then section them in an ultramicrotome, collect on fine-meshed grids, pop in the TEM, and ---presto, nice uniformly thin fiber cross-sections held together by the resin. There will undoubtedly be some tearing of the section and/or individual cross-sections. The fibers will be randomly dispersed so that you'll get many orientations (inlcuding some near-longitudinal slices, if they're useful). Be careful of beam effects - most embedding materials are somewhat to terribly beam sensitive.
Sound too easy to be true? You're right, ultramicrotomy is somewhat of an art, but can accomplish wonders in materials science. A good reference is an overview by yours truly:
T.F. Malis and D. Steele, Specimen Preparation for Transmission Electron Microscopy of Materials, MRS vol 199, Materials Research Society (1990) p.3.
If you don't have any MRS Proceedings there, I have a few copies left and can send one, but only if you're seriously interested, as they're in short supply. The people at UMIST in the Corrosion Center are experts in this area as well. I tend to act as a information bank for this technique, so let me know how you fare - I'm always looking for new references to quote.
Tom Malis Group Leader - Materials Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) Ottawa, Ontario ph.: 613-992-2310 FAX: 613-992-2310 e-mail: tom.malis-at-cc2smtp.emr.ca ____________________________________________________________________________ This may or may not be applicable, but with the high-voltage EM (lower interaction cross-section) and very low beam currents, we have been able to image non- or poorly-conducting specimens without charging or heat- ing problems. If this holds true for your instrument (e.g. you have an IVEM), the advantage is that there may be no preparation required; the disadvantage is that with these low doses you will either need a very sen- sitive recording medium--LoDose or other x-ray film, image plates or in- tensified or slow-scan CCD--or long exposures. The large grain or speci- men drift could make the images useless. Good luck. Yours, Bill Tivol ____________________________________________________________________________ I am a little late in responding to your inquiry on ceramic fibers; My apologies. Since some of the other responses mentioned using the wedge technique for preparing thin TEM sections, I wanted to mention that BUEHLER KKB, with whom I am affiliated, is a supplier of a wedge polishing tool such as those mentioned. I previously was Applications Engineer for South Bay Technology who offered the first commercially available tripod based polisher. However, I have since come to work for BUEHLER, LTD in the US, and we have made some changes to the IBM design in order to enhance the ease of producing wedge samples. We also offer a complete line of diamond lapping films for this polishing system.
If you would be interested in more information regarding BUEHLER's MICROPRECISE(TM) Tripoint Polisher, I would be happy to have literature sent to you, and/or have a salesperson contact you. If you would like more information, please feel free to contact me directly by phone, fax or e-mail.
Best regards, Scott D. Holt BUEHLER, LTD. 41 Waukegan Rd. Lake Bluff, IL 60044 USA Phone: (847)295-4546 Fax: (847)295-7942 102467.2752-at-compuserve.com ____________________________________________________________________________
_________________________________________________________________ Ian MacLaren, Telephone: 0121 414 3447 IRC in Materials, FAX: 0121 414 3441 The University of Birmingham, email: I.MacLaren-at-bham.ac.uk Birmingham B15 2TT, England. _________________________________________________________________
} I am trying to find out if other labs have bulk rates for use of their } electron microscopes.
We have contracted for blocks of time, but we are a NY state fa- cility, so commercial users are not encouraged to buy time on the HVEM.
} I need to try to get an idea of how other labs are handling bulk time } purchases from research labs in their institutions.
We do it on a case-by-case basis--sorry I can't be more helpful.
} Is you lab a departmental or institutional facility?
Yes, but we are also funded my NIH as a Biotechnological Resource.
} What level of support do you receive from the } university/college (% of operating budget)?
50% of our support comes from NY state; 50% from NIH.
} Is there a rational basis for your rate structure?
Yes. Since NY is not allowed either to sell services at a profit or to give services away at less than cost, someone must calculate the total cost of operating the HVEM (including bldg maintenance, etc.), and we charge that cost--~$207/hr. A contract for $5000 (which happened within the last decade) will buy ~25 hr. } } For in-house users we } charge $40 per hour for microscope time,
No charge for in-house users or not-for-profit (incl academic) out- side users.
} and at the moment, charge $3,000 per } year for up to two individuals per grant for unlimited access to the } microscopes.
No analogous arrangements are allowed here (to the best of my know- ledge).
} We are heavily subsidized by the various parts of the college (to } the extent of about 80% of our operating budget). We are working on making } the rates more rational without eliminating all activity. } See above for our solutions. Yours, Bill Tivol
Dear Nigel: We have had a similar problem when polymerizing Lowicryl K4M in a UV chamber, probably due to too much heat generated during the polymerization. You can try reducing the intensity of the UV light, or use indirect light rather than direct light (if you have a foil lined chamber, put a foil covered baffle between the light and the specimen). A smaller volume of resin might also help (e.g. a smaller gelatin capsule). What works well for us is to use disposable polyethylene flat molds (available from E. Fullam, I think), overfilled with resin and covered with Saran wrap to exclude air.
Arthur R. Hand Central EM Facility UConn Health Center Farmington, CT 06030 USA
To: Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com}
We do a calculation with WDS data, which calculates the equivalent thickness and micro-grams of a material sitting on a surface. It's written in APL, an IBM programming language which is not used much any more.
We would like to understand the calculation, and possibly use a spread sheet to do the calculations and store the results for future reference. I bet it uses a bunch of ZAF type corrections, and may therefore be very involved, primarily due to the vast number of correction factors needed.
There is a T/N program called TFSS, which may do essentially the same calculation. We could use it on our 5500 system, and I should see if the results are similar between the 2. Still, I'd have the same problem of blindly feeding numbers in, and accepting the results.
Does anyone know of a report that explains how to do this calculation? Thanks,
If you send me your address, we'll mail you the new catalog "The Microscope Book". I think it will be helpful to you, especially if used in conjunction with its Web page: http://www.shore.net/~catalogs.
Call us if you have any questions. And have a great time with your new "dream microscope".
Regards,
Elinor Solit Director of Publications The Cambrex Group
On Thu, 9 May 1996, Eric Chapman wrote:
} I have just obtained my dream microscope and and trying to get set up in my } home for a small lab. I need to purchase a microtome. Also, I would like a } line on glassware and slides. I have the VWR Scientific catalog but was } wondering if there are other vendors I might write to for catalogs. } } If any of you have phone numbers or addresses I would appreciate it. Also, } if anyone has a used microtome for sale I would love to hear from you. } } --------------------------------------------------------------- } Eric Chapman - Eagle River Alaska _/_/_/ _/_/_/ _/_/ } Hemodialysis Patient _/ _/ _/ _/ _/ } _/ _/_/ _/ } Sic Gorgiamus Alles _/ _/ _/ _/ _/ } Subjectamos Nunc _/_/ _/_/_/ _/_/ } --------------------------------------------------------------- } }
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At 21:10 09/05/96 +0200, Garlasco wrote: } Dear All, } } I use LM with polarized light and contrast liquids (potassium iodo mercurate } in water/glycerol) at 200 and 500 X. } For Chrysotile I use also nitrobenzene or mixtures of cinnammic aldehyde and } benzyl alcohol. I have a Chrysotile standard. } My problem is to manage other asbestos fibers, if present. } How I can see the other asbestos fibers: Crocidolite, Tremolite, } Anthophyllite, Amosite and Actinolite ? I don't have standards with those } fibers. } Some of you can help me ? } } Thank you !! } } Renzo Garlasco } } garlasco-at-newsoft.it _____________________________________________________________________________
Dear Renzo,
The current standard, and probably the most widely used method for the identification of asbestos is polarized light microscopy (PLM). The condition of having crossed polars on your LM will allow you to see many of the optical characteristics that you can use to identify the six main forms of asbestos. These characteristics can include: pleochroism, anisotropy, birefringence, extinction angle, and sign of elongation.
To conclusively identify the asbestiform minerals, you should use a special objective with a 'stop' ( a black disc) placed in the objective back focal plane. Such an objective is called a 'dispersion staining objective'. The addition of this 'stop' will allow you to see 'dispersion staining colours'. These colours represent certain wavelengths of light that correspond to the specific refractive indexes of the fibre or particulate in question. This optical staining technique is referred to as 'dispersion staining'.
The refractive index values obtained from the dispersion staining colours are used in conjunction with the other optical characteristics already obtained to identify Chrysotile, Crocidolite, Tremolite, Anthophyllite, Amosite and Actinolite, as well as other fibres and particulate.
Refractive Index liquids such as those made by Cargille Laboratories are used as the mounting medium for PLM identification of asbestos fibres. Based on a preliminary stereoscopic examination with a standard stereoscope in a fume hood, different Refractive Index liquids are used depending on which asbestos type is suspected.
Certified bulk standards of the asbestiform minerals can be obtained from the National Institute of Standards and Technology.
For more information please e-mail me at: phhrich-at-phhenvironmental.com
Just a quick note of caution to add to the thread about preparing SiC fibers for TEM: SiC is hard enough to damage a diamond knife. My experience with microtoming SiC is all negative: it nicked my knife, and the SiC was basically shattered. I would stick to tried and true embedding, dimpling, and ion milling techniques (or is that tired and true?), or tripod polishing.
There are MANY companies where you can obtain catalogs, but some are more convient than others. Some sales reps are more "helpful" than others. What I would suggest, if you can, drop into your nearest college or University and visit in the Biology or like department that uses microscopes. Go to the Department Office, and ask if you could talk to one of the faculty. Most college/University departments are very open to "strange requests."
Explain to her/him that you are an newly starting "amateur" microscopist and you are looking for sources of supply for equipment and miscellaneous supplies. You may be refered to another faculty member who is more their "resident microscopist." I would be very supprized if you did not meet more than one faculty memeber. They have the catalogs, and *know* who is local and gives good service.
You might also find a friendly ear, and see many things you had not thought about.
Enjoy your microscope.
Shalom from Jerusalem, Azriel Gorski Head, Optical Microscopy Laboratory Israel National Police
On Thu, 9 May 1996, Eric Chapman wrote:
} I have just obtained my dream microscope and and trying to get set up in my } home for a small lab. I need to purchase a microtome. Also, I would like a } line on glassware and slides. I have the VWR Scientific catalog but was } wondering if there are other vendors I might write to for catalogs. } } If any of you have phone numbers or addresses I would appreciate it. Also, } if anyone has a used microtome for sale I would love to hear from you. } } --------------------------------------------------------------- } Eric Chapman - Eagle River Alaska _/_/_/ _/_/_/ _/_/ } Hemodialysis Patient _/ _/ _/ _/ _/ } _/ _/_/ _/ } Sic Gorgiamus Alles _/ _/ _/ _/ _/ } Subjectamos Nunc _/_/ _/_/_/ _/_/ } --------------------------------------------------------------- } }
Does anyone have any tips for polishing gold to use as a standard?
I have available only very basic polishing equipment which generally serves our needs for polished rock sections very well using diamond paste of varying grades.
My problem seems to be that the gold is so soft that when I jump from one micron diamond to 0.25 I only make things worse.
Any advice would be most welcome.
Robert McDonald Geology & Applied Geology Dept Glasgow University Glasgow Scotland
} } Hi, The old Edwards belt driven pumps are heavily constructed and run at a } lower rotational speed than the new direct-drive units. This is sort of } typical of the new construction philosophies. I repair older Philips } Electron Microscopes so I have quite a bit of first-hand experience with } the older pumps. They are rugged but do not always maintain an optimum } pumping speed. Most people don't notice a decrease in pumping speed, so } they happily let their old pumps chug along for decades. I have replaced } about six belt-driven units with the newer, direct drive pumps but I } certainly don't expect them to last as long as the old ones did. The higher } speed, lower volume of oil and thinner castings all contribute to the } longevity problems. } Dear Alex, We have TC gauges on our belt-drive pumps, and the ultimate vacuums for backing the turbopumps (~5 microns) and roughing the column (~50 microns) have not changed for 15 years. We haven't measured the pumping speed direct- ly, but the pumpdown times for the column and accelerator do not seem to have changed. Yours, Bill Tivol
To find straight lines and circular objects one can apply the linear and circular Hough Transform. This transform is not present in my 'C' based IP package (SCIL-Image).
Question :
Does anyone of you know good comprehensive (extensive) literature refs on this subject from which I can write the Houigh Transform programs or even better ...... is there someone out there who has the C routines to calculate the lin. & circ. H.T. and is prepaired to grant them to me ?
I am grateful for any help !
Kees.
BTW. The Image Processing Handbook does not cover the subject in depth in order to write my own programs.
-- Kees van der Wulp
TNO - Prins Maurits Laboratory INTERNET : vanderwulp-at-voeding.tno.nl Division 1 New VOICE : +31 15 2843101 Department : ATES New FAX : +31 15 2843963 PO-Box 45 General TNO Info : http://www.tno.nl 2280 AA RIJSWIJK (NL) THE NETHERLANDS
In reply to Renzo Garlasco's request for info on asbestos standards:
There are two National Institute of Standards and Technology Standard Reference Materials (SRM) specifically designed for the polarized light microscopic identification of asbestos, unluckily they are not free.
Here is a brief description of the materials, so that you can see if you might be interested:
Standard Reference Material 1866 "Common Commerical Asbestos" consists of chrysotile, Amosite, crocidolite, and a glass fiber blank and Standard Reference Materials 1867 "Uncommon Commercial Asbestos" consists of actinolite, tremolite, and anthophyllite. The optical properties and the asbestiform nature of the asbestos materials are certified. The certified optical properties include morphology, pleochroism, birefringence, extinction, sign of elongation, and the refractive indices of the material as a function of orientation and wavelength. Crocidolite is an exception, because of its strong absortion, refractive indices are reported but not certified. You get several grams of each standard.
You can contact me for further technical information on the standards or for information about obtaining the standard you may contact:
Standard Reference Materials Program National Institute of Standards and Technology Gaithersburg, MD 20899-0001 USA
Eric B. Steel, Leader e-mail: eric.steel-at-nist.gov Microanalysis Research Group Office: 301-975-3902 N.I.S.T. FAX: 301-216-1134 Bldg. 222/Rm A113 Gaithersburg MD 20899
We also use diamond for the polishing. The cloth used we have found to be critical. You must use a soft one like Buehler's "Microcloth" with very light pressure. Nylon produces unacceptable results.
Joe Geller Geller MicroAnalytical Laboratory 426e Boston St. Topsfield, MA 01983 jg-at-gellermicro.com
Manufacturer's of gold standards, and other materials, for x-ray microanalysis.
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-- [ From: Charles A. Garber., Ph.d * EMC.Ver #2.10P ] --
In resonse to ======= } Could you please tell me how to well prepare for a TEM sample of SiC fibers in order to observe their cross-sections??? ========== There was quite a long list of extremely valuable information which we found very useful. Thank you!
However there was no discussion on the technical merits of a "tri-pod" polishing/ion milling vs. diamond knife thin sectioning approach. I came to realize early on that nothing in the world of EM is without artifacts, and the challenge more often than not is to make judgements as to what is real and what is artifact. So I was wondering how those with the experience, and those able to really compare the two methods from first hand experience would comment on the matter of artifacts and to what degree one approach is more artifact free than the other.
It has been our own experience, though I suspect very limited in comparison to others who made postings, that the ion milling induced artifcats are isotropic and therefore not so easy to spot where as the ultramicrotomy induced artifacts are anisotropic in nature and to do an "artifact test" one need only to rotate the sample in the microtome and see whether the suspected artifact feature does or does not rotate in the final image. So even though ultramicrotomy might in fact not have fewer artifacts, those that are there are more readily recognizable as to what they are.
Also, in response to the following by Dan Schwartz: =================== Just a quick note of caution to add to the thread about preparing SiC fibers for TEM: SiC is hard enough to damage a diamond knife. My experience with microtoming SiC is all negative: it nicked my knife, and the SiC was basically shattered. I would stick to tried and true embedding, dimpling, and ion milling techniques (or is that tired and true?), or tripod polishing. ===============================
Well it is a fact that cutting "hard" materials like SiC fibers will more quickly wear down a diamond knife in the same way as quick starts and stops will more quickly wear out automobile tires. However, in some instances, diamond knives may in fact be a viable alternative, if not in some cases, a more attractive alternative. True there is cost but the real issue is the cost per sample possible per knife rather than the cost of the knife itself. "Perfection of the art" enables one to obtain acceptable sections faster using fewer passes of the sample over the knife (e.g. less knife wear), and of course using "materials" instead of "life" science diamond knives reduces the per sample cost still further, and by more than a trivial amount. In fact in our own laboratory we have got it down to the point whereby such a "hard" sample adds typically to the client cost an incremental amount not exceeding $150 per sample assuming there are multiple samples. But for the uninitiated, that cost could be far greater. We know of some users who have literally "broken" the knife the first time out.
Disclosure: SPI Supplies offers "Materials Science Diamond Knives" (see on-line electronic catalog) and surely would like to see more diamond knives being used up in the conduct of this kind of work! Also, our analytical services laboratories will thin section "hard" samples for clients as a service.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: spi2spi-at-2spi.com
To add to what Chuck Garber has said re: the identification of artifacts in microtomed vs. ion milled samples:
-------------------------------------------------------------------- "that the ion milling induced artifcats are isotropic and therefore not so easy to spot where as the ultramicrotomy induced artifacts are anisotropic in nature and to do an "artifact test" one need only to rotate the sample in the microtome and see whether the suspected artifact feature does or does not rotate in the final image. " ----------------------------------------------------------------------------
the fracture behaviour of crystalline materials during microtomy can sometimes be used for a quickly identification of phase distribution in a mixed sample. I've used this while studying activated alumina pellets, graphite anodes, and while I've never confirmed it, I've become sufficiently familiar with the fracture artifacts of intermetallic constituents in Al alloys to usually have a good idea what they are by their morphology in the section.
As for which method I choose when preparing a sample (fibre or otherwise), it depends on what I'm looking for, how much time I have, and where the results are going to end up. Regardless of how clearly and quickly my microtomed "fragment" may have shown the thickness of a surface layer, if an image is required for publication, I'm likely to use an ion milled sample. If I'm concerned with quick results where EDXS is required, then microtomy will likely be my method of choice.
Does anyone out there know how to make high-quality overheads from standard B&W photographs (e.g., TEM images including HRTEM images)? I have always made slides for my talks up until now. Just using the standard xerox transparencies does not give very good results, but I have seen talks where people use transparencies and they are remarkably good. Is there some trick or method I am missing out on?
Roy Christoffersen Texas Center for Superconductivity 3201 Cullen Houston, TX 77204-5932 roy-at-bayou.uh.edu (713) 743-8273 FAX: (713) 743-2787
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A San Diego, California based manufacturer of monochrome digital RS-422 (10 or 8 bits) and RS-170 (analog 10 bit video cameras) with S/N of } 62dB in real time 30 frames/ second, with a defect free sensor and 100% fill factor, has a new and informative web site! -------------------------------------------------------------------- (((((((( The URL is: http://www.edt.com/dvc/dvc.html )))))))))) -------------------------------------------------------------------- The company can in addition provide complete (plug and play) frame grabbers for digital or (analog 10 bit / 1024 gray level PCI bus), Mac Nu-bus or, Sun S-bus. SGI digital output option available, as seen on new Hot Mix 11 html SGI CD, along with custom cables, Mac and Sun imaging software and tunable electronic filters for mono 400-1100nm range and (RGB sequential version.) Feel free to e-mail requests directly, stating bus of interest, digital or analog video interest, and application. Signal to noise / pixel gray level depth is just as, or more important than just spacial resolution! Is your camera offering only 7 bits/64 gray levels, or 50dB S/N at best, and a New England snow storm when the gain is turned up? Are you facing the expensive, only 8 bit, ticking 7.5 fps slow expensive digital cameras along with no RS-170 analog output. Full details on how to choose a CCD camera and various frame grabbers listed by bus on the above web site. Feel free to email your application, spectrial nm area of interest etc. Thank you.
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Thanks for all the references, they will help in the quest. However, the quest has not been completed.
What happened to the people who replied when I said that double labeling using two mouse monoclonals had probably not been done before. I received some interesting, and sharp, replies with some good examples of how two monoclonals had been used to label the same tissue slices.
Best regards,
Paul Webster Center for Cell Imaging Yale University School of Medicine http://info.med.yale.edu/cellimg
I am trying to locate information on a stereomicroscope, Model M650, manufactured by Wild-Heerburg, serial no. 10682. Can anyone provide the phone number of a dealer so I can inquire about parts? Many thanks.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
Does anyone have any tips for polishing gold to use as a standard?
I have available only very basic polishing equipment which generally serves our needs for polished rock sections very well using diamond paste of varying grades.
My problem seems to be that the gold is so soft that when I jump from one micron diamond to 0.25 I only make things worse.
Any advice would be most welcome.
Robert McDonald Geology & Applied Geology Dept Glasgow University Glasgow Scotland
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Regarding my previous query: replacing LaB6 e-gun assy with W ... I have just re-installed the LaB6 (currently pumping down), and have inspected the W assy which wasn't working. It is definitely open, but I wondering if what I'm seeing isn't as much an answer but rather a clue. That is, the W filament appears to have been "zapped" ... open with beads on both ends. Could the LaB6 configuration have put too much current through the W filament?? TIA (again) ...
cheers, shaf {\/} /\ {\/} /\ {\/} /\ {\/} cognito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/epmahome/
Robert McDonald requested information on polishing gold by hand or with basic equipment. My suggestion is to perform your final, deformation removing step, on a soft cloth such as BUEHLER's MICROCLOTH(R) or MASTERTEX(R). Instead of 0.25micron diamond, use MASTERMET(R) final polishing suspension with light pressure to polish the gold surface.
The ideal is to perform final polishing on a vibratory polisher such as BUEHLER's VIBROMET(R) 2. Recent years have seen a great change in vibratory polishing at BUEHLER. In the past, vibratory polishing was used sparingly by many people due to the rounding effects incurred. However, the VIBROMET(R) 2 has been changed to produce less of a vertical movement component, and to increase the horizontal component. This effectively eliminates the rounding effects which were the bane of vibratory polishing in the past.
If you have any further questions regarding gold polishing or vibratory polishing, please don't hesitate to contact me (I will be out for the remainder of the week), or one of our other sample preparation laboratory experts by calling (800)BUEHLER [(800)283-4537]. Or if you would rather contact us by FAX: (847)295-7942.
Best regards, Scott D. Holt BUEHLER 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-4546 http://www.buehlerltd.com
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I have removed my LaB6 assembly from our JSM-6300 for cleaning, and wanted to replace it with, and use, the W assy in the mean time. The trouble is the HV heat indicate some amount of the "preheat" which is for the LaB6 but I can't put any more current thru the filament than that ... and the emission implies zero. The logical thing to check is if the filament is actually open but it should be good.
It is my impression in spite of the LaB6 demanding "preheat", I should be able to interchange both gun assemblies. Is there something which is associated with the LaB6 configuration that I am missing??
Also: in spite of you 840|6300 users not be able to help, please respond anyway (directly ... see "oregon" address below) ... I'd like to create an e-mail group which is a bit more specific to my "microscopy" applications. TIA ...
cheers, shaf {\/} /\ {\/} /\ {\/} /\ {\/} cognito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/epmahome/
We have a FuJix color printer that does transparencies very well. It is not cheap and the transpareancy paper itself is ca. $3.0o a page. The Codonics printers also do a good job. They are ca. $9000. Good luck. Nina allen
On Tue, 14 May 1996, Roy Christoffersen wrote:
} Does anyone out there know how to make high-quality overheads from standard } B&W photographs (e.g., TEM images including HRTEM images)? I have always } made slides for my talks up until now. Just using the standard xerox } transparencies does not give very good results, but I have seen talks where } people use transparencies and they are remarkably good. Is there some trick } or method I am missing out on? } } Roy Christoffersen } Texas Center for Superconductivity } 3201 Cullen } Houston, TX 77204-5932 } roy-at-bayou.uh.edu } (713) 743-8273 } FAX: (713) 743-2787 } } }
} Does anyone have an idea how long and how fast to spin IgG gold } solutions and BSA to help reduce background on en-grid imunogold } labelling. } } I am specifically looking at p450 enzyme in lung tissue but am having } a bit of difficulty with high backgroud in my controls. } } Thanks alot } } Ray Gilbert } University of Leicester } e-mail rtg2-at-le.ac.uk } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
Depends on the size of the gold. we spin 15-20 nm gold one minute at top speed on a microfuge. As our IgG gold gets older a small pellet will form. New stuff rarely has a pellet ******************************************************* Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
Dear Paul Webster, I did not see the messages that you were talking about. But I remember reading about this in an article by A.C. Cuello in the book Immunochemistry II, that he/she edited, published in 1993, by Wiley, in their "IBRO Handbook Series: Methods in the Neurosciences (General editor A.D.Smith) Volume 14. The book is mostly about how to follow neurons around in huge volumes of brain, but Cuello has an article that deals with multiple mab labelling. If I remember, the method required direct conjugation of different dyes to the different mabs. This sounds a bit tedious, but then it has the delightful advantage of elimanating hassles with crossreacting secondaries. I have no personal experience with the method.
Here is a brief summary of the responses to my question of last week:
} has anyone } besides me had problems with osmium made up in PIPES?
I am not sure whether their responses went to the List or just to me, but my thanks to Rosemary White, Tom Phillips, Larry Oakford, and John Heckman for their comprehensive replies. Sorry I don't have the time to respond to each of you separately.
Rosemary suggested the problem was with the NaOH used to adjust the pH of the PIPES, which makes sense since that was what I used. She recommended KOH-PIPES or a mix of MES & PIPES instead.
Larry considered Veronal buffer to be the most appropriate buffer for OsO4, but the majority opinion (LO, TP, JH) was that unbuffered osmium works at least as well as buffered osmium.
Coincidentally, I discovered that our trainee technician recently made up our osmium unbuffered .... and no-one noticed. (Don't tell our Linda though!)
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department School of Medicine University of Auckland Private Bag 92019 Auckland New Zealand
The main methods of producing high-quality EM viewgraphs that I've come across are:
1. Going to a Copyshop that has a Laser Photocopier, viewgraph ready for photocopying, and having it lasercopied onto transparency. The results are remarkably good in my experience; because the laser-copier prints digitally you can also photocopy a laser-copy with results far better than a photocopy of the original (e.g. for hand-outs from your talk etc.).
2. We obtain very nice results by scanning the original EM negative into a PC using an AGFA Arcus II flatbed scanner; the picture files are then inverted (to get a positive), imported into MS Powerpoint and incorporated in viewgraphs that way. The scanner is used in 'transmission mode' for negatives. If you have a scanner without the transmission feature, scanning a print works almost as well for viewgraphs.
} } Does anyone have an idea how long and how fast to spin IgG gold } } solutions and BSA to help reduce background on en-grid imunogold } } labelling. } } } } I am specifically looking at p450 enzyme in lung tissue but am having } } a bit of difficulty with high backgroud in my controls. } } } } Thanks alot } } } } Ray Gilbert } } University of Leicester } } e-mail rtg2-at-le.ac.uk } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } Depends on the size of the gold. we spin 15-20 nm gold one minute at top } speed on a microfuge. As our IgG gold gets older a small pellet will form. } New stuff rarely has a pellet } ******************************************************* } Greg Erdos Phone: 352-392-1295 } Scientific Director, } ICBR Electron Microscopy Core Lab } 218 Carr Hall Fax: 352-846-0251 } University of Florida E-mail: gwe-at-biotech.ufl.edu } Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ } } ******************************************************* } } ******************************************************* Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
Joe Michael writes, } Dear Colleagues, } } There is an excellent reference on the Hough Transform that covers both the } extraction of linear features and circular features. The title is } } "Shape Detection in Computer Vision Using the Hough Transform" } } by V. F. Leavers } Springer-Verlag } New York
For your information, Visilog from Noesis Vision Inc provides a module for doing Hough Transform and Correlation. You can extract linear or circular features using these algorithms but also predefined shapes in an image using a model of the shape.
---------------------------------------------------------------------------- --------------------- Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com http://www.cam.org/~noesis 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada
Join the Noesis NewsGroup at Visilog-at-noesisvision.com ---------------------------------------------------------------------------- ---------------------
} Date: Tue, 14 May 1996 08:53:53 -0500 } From: (Roy Christoffersen) {roy-at-bayou.uh.edu} } Subject: High-quality overheads from EM micrographs } To: Microscopy-at-sparc5.microscopy.com } MIME-version: 1.0 } Content-type: text/plain; charset="us-ascii" } Content-transfer-encoding: 7BIT } } Does anyone out there know how to make high-quality overheads from standard } B&W photographs (e.g., TEM images including HRTEM images)? I have always } made slides for my talks up until now. Just using the standard xerox } transparencies does not give very good results, but I have seen talks where } people use transparencies and they are remarkably good. Is there some trick } or method I am missing out on? } } Roy Christoffersen } Texas Center for Superconductivity } 3201 Cullen } Houston, TX 77204-5932 } roy-at-bayou.uh.edu } (713) 743-8273 } FAX: (713) 743-2787
Roy,
You can digitize you EM images, import into PageMaker, export to high quality laser printer (} 1200dpi), and print on transparencies directly. The quality is much better than from xerox. Good luck,
Ya Chen
Ya Chen
========================================================================== Cryo/SEM Coordinator Integrated Microscopy Resource (IMR)-- III M M RRRRRR an NIH Biomedical Research Resource I M M M M R R University of Wisconsin, Madison, WI I M M M RRRRRR 1675 Observatory Drive #167 I M M R R Madison, WI 53706, USA I M M R R TEL : 608-263-8481 I M M R R FAX : 608-265-4076 III M M R R Email:YChen-at-macc.wisc.edu ========================================================================== IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html 2nd Symposium on Integrated Microscopy: Sept. 20-22, 1996
Thank you to everyone who replied to my query on differentiation live and dead plant cells.
The approaches fell into two categories.
Firstly those detecting membrane integrity using dye exclusion (eg Trypan Blue) sometimes with a stain such as Neutral Red that is taken up by the live cells.
Secondly those detecting enzyme activity in the living cells, particularly using fluoroscein diacetate (non-fluorescent) which is split by esterases in living cells an then fluoresces. Example references
The Heslop-Harrison & Heslop-Harrison (1970) Stain Tech. vol 45 pp 115- 120 - pollen. For a tissue culture reference see Widholm, J.M. (1972) Stain Tech. vol47 pp 189-194. Harris, N and Oparka, K - Plant Cell Biology, A practical approach (1994) IRL Press
A modification of this second is using propidium iodide to penetrate dead cells - (live green fluorescence, dead red - using blue excitation)
In a similar vein the use of LIVE/DEAD kits from Molecular Probes was suggested by several people though these are not designed for plant cells.
We have carried out an initial test using fluoroscein diacetate which has worked in our experimental system, though there are some problems with clumping of the cells.
Once again thanks to everyone
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
Thank you to everyone who replied to my query on differentiation live and dead plant cells.
The approaches fell into two categories.
Firstly those detecting membrane integrity using dye exclusion (eg Trypan Blue) sometimes with a stain such as Neutral Red that is taken up by the live cells.
Secondly those detecting enzyme activity in the living cells, particularly using fluoroscein diacetate (non-fluorescent) which is split by esterases in living cells an then fluoresces. Example references
The Heslop-Harrison & Heslop-Harrison (1970) Stain Tech. vol 45 pp 115- 120 - pollen. For a tissue culture reference see Widholm, J.M. (1972) Stain Tech. vol47 pp 189-194. Harris, N and Oparka, K - Plant Cell Biology, A practical approach (1994) IRL Press
A modification of this second is using propidium iodide to penetrate dead cells - (live green fluorescence, dead red - using blue excitation)
In a similar vein the use of LIVE/DEAD kits from Molecular Probes was suggested by several people though these are not designed for plant cells.
We have carried out an initial test using fluoroscein diacetate which has worked in our experimental system, though there are some problems with clumping of the cells.
Once again thanks to everyone
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
A high background is unlikely to be fixed by centrifugation. Rather look at your method. Possible causes include (partly taken from BioCell's Gold Conjugates Technical Information and Guidelines booklet, partly personal experience and partly a selection of papers):
..Ionic concentration too low. Use increased salt concentration, up to 30%. ..Inadequate washing. ..Nonspecific charge attraction of antibody. Use detergent (I use 0.4% saponin; Tween is OK). Include normal serum (from the species the gold conjugate came from) at 2% and cold water fish gelatin at 1% in antibody solutions and at 10% and 1% respectively in the preincubation solution. ..Free aldehyde groups in tissue. Float sections on 0.5M NH4Cl for 1 hour before incubation. ..Antibody concentration too high. ..Gold conjugate concentration too high. ..Tissue poorly fixed. Damaged cells stain nonspecifically.
Is the background on particular cell structures (eg nuclei), is it all over the tissue containing area, or is it on the whole section (including resin only areas)? Look at this first. Is the antibody or gold conjugate OK or has it been around too long? Is there any bacterial contamination? What sort of water are you using? Has the method worked before? Has the weather changed recently; were you in a bad mood; was the moon full? Good luck.
Diana van Driel Dept Ophthalmology Sydney University AUSTRALIA 2006
I have had some reasonable results printing digital images onto overhead transparencies on a 600 dpi laser printer. You should scan the images in to at least 2000 by 2000 pixel resolution. The quality depends upon the original digital image and the printer, so the better video printers will do a better job. Also, some copiers have a photo copying option (gray-scale) that will reproduce mid-tones.
However, I don't think an overhead transparency will ever be as good as a slide. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
} Hi All in Microland: } } Does anyone have any tips for polishing gold to use as a standard? } } I have available only very basic polishing equipment which generally } serves our needs for polished rock sections very well using diamond } paste of varying grades. } } My problem seems to be that the gold is so soft that when I jump from } one micron diamond to 0.25 I only make things worse. } } Any advice would be most welcome. }
Dear Robert
The cloth is important as mentioned already as pressure. Having absolutely clean uncontaminated cloths are important as well as moving the sample in a spiral movement on the cloth in order to ensure a even distribution of the diamonds. To prevent cross contamination there should not be gap between the sample and the mould. Araldite is brilliant and is available in most EM labs. Reasonable stable under the beam when slightly C-coated. To ultrasonic clean the sample between cloths with sample face down is also a plus to prevent cross contamination. For final polish of 0.25 micron I had better luck with Cerenium Oxide than with diamond. Stephan H Coeztee Electron Microscope Unit Private Bag 3 Wits 2050 South Africa
Via: uk.ac.bbsrc; Wed, 15 May 1996 09:16:42 +0100 X400-Received: by /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Wed, 15 May 1996 09:17:49 +0000 X400-Received: by mta arcs.bbsrc.ac.uk in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Wed, 15 May 1996 09:17:49 +0000 X400-Received: by mta LARS.MUAS in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Wed, 15 May 1996 09:02:09 +0000 X400-Received: by mta LARS in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Wed, 15 May 1996 09:02:13 +0000 X400-Received: by /PRMD=UK.AC/ADMD= /C=GB/; converted (ia5-text); Relayed; Wed, 15 May 1996 09:02:18 +0000 Receipt Notification Requested) (IPM Return Requested)
Dear Fellow Microscopists,
Just a short note to thank all those who kindly replied to my original query about problems with bubble formation in methacrylate. I have received lots of ideas of ways to overcome the problem: I will be trying them when I have some more plant material ready. In the meantime, it is always comforting to know that there are people out there willing to share their knowledge. 'A problem shared, is a problem halved, quartered, etc...'
Thank you,
Nigel
------------------------------------------------------------------------------- Internet: nigel.chaffey-at-bbsrc.ac.uk IACR-LARS, Dept. of Agricultural X400:G=nigel; S=chaffey; O=bbsrc; P=uk; C=GB Sciences, University of Bristol, Tel: +44 (0)1275-392181 ext:230 Long Ashton Research Station, Fax: +44 (0)1275-394281 Long Ashton, Bristol, BS18 9AF -------------------------------------------------------------------------------
"A high background is unlikely to be fixed by centrifugation."
Unfortunately, we have just discovered that ultra-high centrifugation of some antibodies prior to labeling, can remove non-specific binding on sections. I have no explanation for this and offer my apologies for complicating the field further.
There is a section on our CCI home page concerning problems with antibody labeling if anyone is interested.
Thank you to all those who re-posted the information on muliple antibody labeling protocols using monoclonal antibodies.
Paul Webster Center for Cell Imaging Yale University School of Medicine htt://info.med.yale.edu/cellimg
Thank you for the information concerning the different types of microtomes. My colleague much appreciates it.
Sincerely,
Ginger R. Baker EM Lab Manager Dept. of Anatomy, Pathology, and Pharmacology 250 Veterinary Medicine Oklahoma State University Stillwater, OK 74078 (405) 744-6765 FAX: (405) 744-5275
Thank you for the information concerning the different types of microtomes. My colleague much appreciates it.
Sincerely,
Ginger R. Baker EM Lab Manager Dept. of Anatomy, Pathology, and Pharmacology 250 Veterinary Medicine Oklahoma State University Stillwater, OK 74078 (405) 744-6765 FAX: (405) 744-5275
I am seeking information on the requirements to digitize our Hitachi S-2400 SEM. We would like to digitally capture, archive (on CD-R, etc.), and analyze images on a Power Mac. Can anyone recommend a good frame grabber interface? Would we need any other equipment/hardware? I would also appreciate any price information as we need this for a budget revision on a pending NSF grant, which is due in a couple of days.
Thanks in advance.
Stephen Beck Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829
{I have made a diblock copolymer and would like to investigate microphase separation behavior in this material. One of the blocks is PMMA while the other one is a linear conjugated polyene. I think osmium tetroxide is a good first choice but I need some advice as to the exact way to stain. I already have the samples in the grids. Do I stain with the vapor from an aqueous solution or do I put the grids directly into the solution? for how long? Please reply to tmmaddux-at-midway.uchicago.edu Phone 312-702-1147 Todd MADDUX
I have made a diblock copolymer and would like to investigate its microphase separation behavior using TEM. One block is PMMA while the other is a linear conjugated polyene. I think osmium tetroxide is a good first choice for staining but I really don't know how to go about it. The sample is already on the grids. Should I expose the grids to the vapor from an aqueous solution or should I place the grids directly in the aqueous solution. For how long? Please reply to TMMADDUX-at-MIDWAY.UCHICAGO.EDU
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Hi,
I am looking into the purchase of a new conventional (W/LaB6) SEM for a Materials Science laboratory and had a couple of questions for users of newer SEM equipment: 1. I was told that LaB6 doesn't buy much of a brightness improvement over W and therefore isn't worth the added expense, is this your experience? 2. With digital imaging as the fast increasing method for documentation, is the record/film option still necessary?
All inputs are greatly appreciated as well as any other comments along these lines. Thanks in advance.
I believe there were some recent posts about capturing images of microscope slides on a scanner. Unfortunately I did not save them but now I find that this might be just what we need to do. Could someone briefly summarize what was learned and respond either on the list or to me privately?
Gary Radice 804-289-8107 (voice) Department of Biology 804-289-8233 (FAX) University of Richmond Richmond VA 23173 USA
Hi! If you are making overheads from negatives, Ted Pella has a Dry Silver Processor made by 3M that makes excellent overheads. You expose the negative on the overhead transparency on the enlarger the same as if you were printing the negative. It then takes 6 seconds to run it thru the processor for the finished product. The printer sells for $2150. The overhead transparency film costs about $2 apiece.
Lee Dickey, Micro Lines Marketing Representing: Ted Pella Inc., RMC Inc., Denton Vacuum, Durst, and PGT
I'm back again! Thanks for all the previous help. I love this place!!
Today's problem involves a way to increase gold staining. I have tissues freshly fixed in 2% PFA (also long term fixed in PLP) and run up in both LRWhite and Lowicryl K4M. Can these resins be etched to reveal more structure? Does the antibody and gold label get much below the surface anyway? I have increased time in primary to 24hrs. and don't get much staining. The P.I. on this project has great LM under fluorescence so we know that the antibody works. I am working with a kit from E.Y. Laboratories for the other solutions. Any and all help is deeply appreciated.
Does anyone know who makes attachments/accessories which add confocality to existing microscopes?
I have read that a spinning disk with pinholes may be used at the focus of a 160 mm conjugated microscope lens to provide a confocal arrangement that can work with a camera at video rates.
I have a home-built inverted microscope and want to add confocality, but require video rates (I use an image intensified camera to look a fluorescently tagged DNA in solution in real-time).
I would like to thank everyone who responded to my question about the MT 7000 ultramicrotomes.
I got a number of responses that ranged from very happy to very dissatisfied.
In the interim, I have been in contact with RMC and they have been extremely helpful in trouble-shooting the problems and following up with service. Specifically, I would like to thank Mr. Greg Carter and Mr. Tom Kennedy, who both have given me considerable attention and assured me that my problems with these instruments will be resolved. We just need to wait a bit now, for parts..
I am finding this forum quite useful to solve issues that probably occur in several labs. Thanks again!
Christine H. Block, Ph.D. VA Medical Center Cleveland, OH
-- _____ |\___\ || | \|___| mow down MO'TOWN--GO TRIBE!
First assumption: your Hitachi 'scope has a conventional video out (either NTSC or PAL) port. If that assumption is invalid, contact 4pi analysis about a full digital interface for running the 'scope.
The Scion LG-3 frame grabber is about $900 and does an excellent job digitizing images with the NIH Image software. One caveat - make sure you have a full-size (12") PCI or NuBus slot in your PowerMac (the 6100, for example, has a 7" slot). Be sure to specify video format (US video = NTSC/RS-170, European video = PAL/CCIR).
We use both systems regularly and are pleased with the performance.
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
Scion Corporation (frame grabber) 152 West Patrick St. Frederick, MD 21701
voice: 301-695-7870 fax: 301-695-0035
4pi Analysis (digital EM control and EDS capture) 3500 Westgate Dr Durham, NC 27707-2534
Hello, I am trying to bring an old ISI SX-30 Scanning Electron Microscope back to usable condition and have great difficulty trying to find good documentation and usable schematic diagrams. I realize this is an old instrument and maybe not exactly wonderful when it was at it's best. Topcon Technologies (the new owners of ISI) have been contacted but offer little help, even though we were willing to pay for the necessary documents. One thing we did buy was what appears to be about a 3rd or 4th generation Xerox copy of the system schematics. It is very difficult to read. Any help would be greatly appreciated.
I would appreciate any feedback from people using fluorecsent antifade compounds,such as DABCO,p-phenylenediamine or commercial kits such as slowfade , prolong, citifluor etc.Being relatively new to the wonderful world of fluorescent microscopy, I'd like to know how good they are,and are they easy to use. Thanks
We are processing insect eggs for TEM and SEM. The egg is covered with a impermeable surface that does not allow for the transfer of solutions and especially resins. One attempt has been made following the standard protocol which resulted in a full collapse of the sample. Cutting a slit in the egg is not possible because the inside flushes out. Does anyone know of a method for making this surface permeable?
Thank you in advance
---------------------------------------------------------------------------- --------- Karen Vaughn Tel.(904) 392-1184 EM Technician University of Florida Fax.(904) 846-0251 Electron Microscopy Core Laboratory Email. KLV-at-biotech.ufl.edu Interdisciplinary Center for Biotechnology Research http://www.biotech.ufl.edu/~emcl/ 214 Bartram Hall Gainesville, Fl 32611
I got several responses to previous BB postings asking how to get access to our differential contrast (hysteresis) imaging software (detailed in MSA and MAS 95, 96, Scanning 1996, and my WWW pages).
We now have for distribution a software version for Silicon Graphics Workstations (using SGI libraries) for 8-16 bit graytone, 24-bit RGB color (both as TIFF), and unlimited image sizes. Since differential hysteresis processing is very computer intensive, we are developing an option for networking several SGI workstation for image processing through distributed computing. A single SGI Indigo2 will need ca. 15 min. for a 8-16 bit 1Kx1K image, graytone or color. Smaller images require fractional time (256x256 24-bit Color Tiff less than a min.).
The software is described at: http://panda.uchc.edu/htklaus/DHP-Img.html; graytone applications are at: http://panda.uchc.edu/htklaus/Microsc-Img.html; 24-bit color application are at: http://panda.uchc.edu/htklaus/Color/Color-Intro.html.
If you like to use this software please use the form provided at http://panda.uchc.edu/htklaus/DHP-Img.html (if you do not have web access please let me know by return email). You will need a SGI workstation, you must work at an academic institution, and you will have to sign an University license for non-distribution and non-commercial use, because my University patented the technology. However, the processing results--I am sure--will be an adequate reimbursement for the licensing inconvenience.
Also, we are putting the software on the web for unrestricted interactive image processing (will be available in a few months).
Does anyone know of a software package which will allow measurements of rotary shadowed molecules traced on a digitizing tablet from TEM micrographs? We do not want to take the time to scan the images into the computer, and DO want to work with printss. The molecules are quite kinked, so measurements with a loupe are problematic. At one time we had a Bioquant system, using a digitizing tablet and an IBM PC XT, but the tablet is broken and no longer available, and the software worked only with that tablet. It did everything we wanted when it worked. It would be nice if the suggested software would work on a IBM computer and an off the shelf digitizing tablet, though MAC based software would also work. We already have NIH Image, but the act of scanning hundreds of images into the computer is not very unattractive.
Yes I have seen the recent rash blatant advertising on the listserver and have sent messages to each of the offenders. Please be advised that none of these posting came with my "approval" even though at least one was suggested by it's leading message. In addition 2 of these individuals are not subscribers to the distribution list, they were simply opportunists taking advantage of the Email system. Unfortunately after sending a rebutting message there is little I can do.
This is an unmoderated server and as such I see the messages at just about the same time all of you do. Occasionally I get requests to review postings and will continue to do so and advise on the appropriateness of any posting. Many commerical subscribers take advantage of this and you should all appreciate their co-operation.
Just as a refresher I will repost a portion of the Listserver "rules". I am please to say that the vast majority of you follow them quite well. As always, thanks for your help, it makes my life simplier and allows me to get to sleep earlier.
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Hello All,
I guess this might interest fairly many of you. I have used the method which I learned in Japan, Sendai. Juji makes a product called FUJIGRAPH PROJECTION FILM PT-100, which you can get in A4-size. The film can be used as enlarging paper and the results are very good. They used this film in the MRI in Tohoku University, where Professor Hiraga taught me to use it.
Regards, Jouko
Jouko K. M=E4ki, Ph.D., Laboratory Manager University of Turku, Laboratory of Electron Microscopy Kiinamyllynkatu 10 FIN-20520 TURKU FINLAND Tel: +358 21 333 7318 GSM: +358 40 505 2521 FAX: +358 21 333 7380
Dear all, Please will you e-mail me protocols for DAB intensification using transition metal salts if you use these methods. I have also put this request out on the confocal listserver. For those of you on both listservers I apologise for the duplication.Jeremy.Sanderson-at-path.ox.ac.uk Jeremy Sanderson Light & Electron Microscopy Sir William Dunn School of Pathology South Parks Road Oxford OX1 3RE Tel: 44-1865 275539 Fax: 44-1865 275515 e-mail:Jeremy.Sanderson-at-path.ox.ac.uk
Neal Nicklaus wrote: } } Does anyone know who makes attachments/accessories which add confocality to } existing microscopes? } } I have read that a spinning disk with pinholes may be used at the focus of a } 160 mm conjugated microscope lens to provide a confocal arrangement that can } work with a camera at video rates. } } I have a home-built inverted microscope and want to add confocality, but } require video rates (I use an image intensified camera to look a } fluorescently tagged DNA in solution in real-time). } } Neal Nicklaus } } SEQ Limited } } Voice: 609-452-6033 Ext. 13 } Fax: 609-452-5955 } email nnicklaus-at-seq.sarnoff.com
Dear Neal,
Nikon, Inc. markets the K2sBio (Nipkow Spinning Disk) Confocal Attachment and is priced at about $16K. Unfortunately, this unit will only work on Upright compound microscopes as it takes the place of a traditional Epi-Fluorescence attachment. To solve your dilemma contact Meridian Instruments at (800) 247-8084, they make a similar unit named the Insight that works on most Inverted platforms.
Good Luck,
Lawrence Kordon Nikon, Inc. Columbia, Maryland nikon-at-jagunet.com
A friend of mine will be offering an EM course at the University of Hartford this fall. The course has not been run for a few years and he finds that he is in need of an ultramicrotome (or two). As is the case with all small colleges, there is not a great deal of money available. He would like to spend as little as possible (free would be ideal) on a ultramicrotome. Since he is most familiar with the Sorval MT2B, he was hoping that someone out there might have one sitting in their lab, in more-or-less working order, that he could have.
He does not have access to the internet, so he asked me if I would send this message out. So, if there is anyone who has a MT2B and would not mind parting with it, please Email me.
Thanks, Marty Levin
Martin A. Levin Department of Biology Eastern Connecticut State University Willimantic, CT 06226 Phone: (860)465-4324 FAX: (860)465-5213
Hello Peter, I use the receipe of Christian Broesamle (tips and tricks in netscape) The receipe is as follows: 6 g glycerol (analytical grad) 2,4 g mowiol 4-88 [Calbiochem #475904] 6 ml dd H2O add 12 ml 0.2 M Tris buffer pH 8.5 and mix for half a day on shaker let the mixture sit for 2h incubate at 50 degrees centigrade for 10 min centrifugate mixture at 5000g for 15' aliquot and freeze supernatant at -20 C until use add 0.1 % DABCO [Aldrich #D2.780-2]
This helps to reduce fading but still.... good luck annette Bakker --------------------------------------
Peter Smith
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Mary Anton wrote: Hi, I am looking into the purchase of a new conventional (W/LaB6) SEM for a Materials Science laboratory and had a couple of questions for users of newer SEM equipment: 1. I was told that LaB6 doesn't buy much of a brightness improvement over W and therefore isn't worth the added expense, is this your experience? 2. With digital imaging as the fast increasing method for documentation, is the record/film option still necessary? All inputs are greatly appreciated as well as any other comments along these lines. Thanks in advance. Mary Anton University of Connecticut USA
My response........... Hi Mary and all,
I'll throw my two cents in! Technical advantages of LaB6 aside, the extended life is worth it, as you'll spend less time changing filaments. To keep a W filament working at peak potential, you nees to spend a significant amount of time keeping the anode, gun and column clean. Also , depending on your SEM, you may need to assure that the W filament is mechanically aligned for best performance. I also think that the W filament is more stable once it's been in use for a while, which is an advantage when doing ultra slow scans required for Xray mapping, SC imaging and some BSE work.
WRT the Polaroid option: If your clients have PC's, I'd say don't get it. Network the images to them and let them view them on the PC. If you absolutely need a film copy at some point in the future, send your image to a printer and have it printed on any media you wish. In my experience a printout (or two) on an inexpensive 600 dpi printer can convey as much information as a Polaroid if the image is carefully chosen. I'm not saying the print quality is as good, but a low mag and possibly a high mag view of the area of interest can convey as much information to the user as a Polaroid.
Please keep in mind that my suggestions are based on a non-existent knowledge about exactly what you'll be doing. In general, I think materials people would agree with me, but I'm very interested in reading their response.
The University of Florida is strongly urging all of us to drop our service contracts on all types of equipment in favor of going with a company called CIC Agency who will handle service payments. We chose the service provider and they pay for the services charges. It sort of like an HMO. We pay a fixed annual fee.
Has anyone had experience with CIC or similar? If so could you share with us your feelings. ******************************************************* Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
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I would like to subscribe to the Microscopy Listserver. My email address is DM-at-lilly.com.
Announcing the Annual Minnesota Microscopy Society Annual SPRING SYMPOSIUM ***********************************************************
To be held at SHERATON INN, MIDWAY, I-94 at HAMLINE AVENUE, ST. PAUL MN on THURSDAY - MAY 23, 1996
*********************************************************** SCHEDULE OF EVENTS 8:00 - 9:00 am Coffee and Late Registration
9:00 - 9:45 am Understanding Video Signals: Cameras, Recording Formats and Printers MARTY HARALDSON, Alpha Video, Edina, MN
9:45 - 10:30 am Coffee Break \ Vendor Displays
10:30 - 11:15 am Capturing, Storing and Organizing Digital Image Files MARK SANDERS, Imaging Center, College of Biological Sciences, University of Minnesota, St.Paul Campus
11:15 - 12:45 pm A buffet lunch will be served at the Sheraton Inn
12:45 - 1:00 pm MMS Business Meeting and Election of Officers
1:00 - 1:45 pm Microscopy Resources on the Internet Dr. STUART MCKERNAN, Center for Interfacial Engineering, Characterization Facility, U of MN, East Bank Campus
1:45 - 2:15 pm Coffee Break \ Vendor Displays
2:15 - 3:00 pm Public Domain Shareware, Microscopy and the Internet Dr. DAVID BRIGHT, 1996 MSA Traveling Speaker Research Chemist, Microanalysis Group, NIST, MD
3:00 - 4:30 pm Vendor Displays
Please make your reservation in advance! NO LATER THAN MONDAY, MAY 20, if you plan to attend the Symposium (or you can pay at the door if unavoidable). Contact Stuart McKernan at (612) 626-7942, stuartm-at-maroon.tc.umn.edu or Dwight Erickson at (612) 736-2830, usmmm214-at-ibmmail.com
Symposium Fee: $20.00 current regular MMS members 95/96, $10.00 student members 95/96, $30 non-member(confers regular membership), or $15.00 non-member students (confers student membership).
Stuart McKernan stuartm-at-maroon.tc.umn.edu CIE Microscopy Center, University of Minnesota Office: (612) 626-7942 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
Message-Id: {199605161453.KAA14180-at-vaxserv} X-Sender: nnicklaus-at-cave.sarnoff.com X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I thank everyone for the feedback. A brief summary is below. I'll send an update as more info arrives.
1) Tracor Ltd in the USA making tandem confocals
2) Noran (http://www.noran.com/)
3) Meridian Instruments at (800) 247-8084, attachment for inverted platforms
4) Nikon, Inc. markets the K2sBio (Nipkow Spinning Disk) Confocal Attachment and is priced at about $16K. Unfortunately, this unit will only work on Upright compound microscopes
5) Possible new design with better light budget by Rimas Juskaitis' and Tony Wilson's
6) Recommended book to read is: (Handbook of Biological Confocal Microscopy, James B Pawley ed. 2nD edition, Plenum ISBN 0-306-44826-2)
--------------------------------------- At 12:13 AM 5/16/96 -0800, you wrote: } } Does anyone know who makes attachments/accessories which add confocality to } } existing microscopes? } } } } I have read that a spinning disk with pinholes may be used at the focus of a } } 160 mm conjugated microscope lens to provide a confocal arrangement that can } } work with a camera at video rates. } } } } I have a home-built inverted microscope and want to add confocality, but } } require video rates (I use an image intensified camera to look a } } fluorescently tagged DNA in solution in real-time). } } } } } } } } Neal Nicklaus } } } } SEQ Limited } } } } Voice: 609-452-6033 Ext. 13 } } Fax: 609-452-5955 } } email nnicklaus-at-seq.sarnoff.com } } } } } Nick, } } I'd be quite grateful if you'd share your results with me. } } Thanks, } } } } } } } Regards, } } } } (signed) Ed Monberg {em-at-mediacity.com} } } -------------------------------------------------- } } 510-429-1060 Fax 429-1065 } LMDC, (Laser Motion Development Co.) } 3101 Whipple Road } Union City, CA 94587-1216 } } } For our Most recent Catalogue of "On Hand" EQUIPMENT: } Send empty mail to: {Cat-at-lasermotion.com} } } } Our web page: http://www.lasermotion.com (Is beginning to take shape!) } Our e-mail: office-at-lasermotion.com } } {-------------------------------- Our page width } -----------------------------} } } } }
I'll answer just one of Mry Anton's questions, that regarding LaB6 cathodes. I tested LaB6 cathodes extensively on our Philips TEMs a few years ago (reported in the 1994 MAS Proceedings). The result is that you'll only see a factor of 3 improvement over W filaments if you buy LaB6 cathodes with a 90 degree cone angle. The 10x brightness which has been touted for years only occurs for smaller cone angles (60 degrees). If one actually looks at the original literature which is the source of the 10x claim (J. Vac. Sci. Technol. 15(3)1978), the cone angle of the emitters is less than 90 degrees.
Russell E. Cook Electron Microscopy Center for Materials Research Argonne National Laboratory 9700 South Cass Avenue MSD-212 Argonne, IL 60439 (708)252-7194 FAX: (708)252-4798
I'm spending more and more time at my keyboard doing images etc and find myself suffering. Someone suggested the ergonomic keyboards have been a real help, in addition to changing desks, layouts, foam pads etc. Anyone have some pros and cons of these odd looking keyboards before I run out and buy one? I have read several books in this regard, but would welcome any other tips on reducing repetitive stress (my doctor, bless him, said dryly, well, if it hurts to do it,....then don't do it....).
thanks
steve
Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University San Diego, CA 92182-4614 phone: (619)594-4523 fax:(619)594-5676 email:sbarlow-at-sunstroke.sdsu.edu
Carl Zeiss has just announced that they are selling their own used equipment. What a chance to get some of those wonderful older microscopes. Suggest you call them at 800-356-1090. Let me know how this works out for you.
Regards,
Ellie Solit The Microscope Book
On Thu, 16 May 1996, SNYDER, JOSEPH wrote:
} I am looking for sources of used equipment such as equipment brokers etc. } Specifically the types of equipment include coating, microscopy and } mechanical test equipment. Secondly I'm also trying to locate sources of } discounted consumable supplies for metallurgical polishing work. } } I would deeply appreciate any leads. }
I use sodium borate, add enough to raise pH to about 9.0. Our chillers are on a preventative maintainance schedule with our maintainance department and they change the water. I just add the borate.
Best of luck Ed Calomeni Medical College of Ohio Toledo, OH 43699 emlab-at-opus.mco.edu
The water in our recirculating unit was recently changed under our yearly service contract. The engineer filled it with water from our Milli-Q system and had us order one liter of Ethylene Glycol (Fisher Scientific) to add to the system. I just checked, and there is nothing growing, that I can see, however there is a faint metallic sheen across surface of the water. Hope this helps. Linda M. Fox lfox1-at-wpo.it.luc.edu
} I was told that LaB6 doesn't buy much of a brightness improvement } over W and therefore isn't worth the added expense, is this your } experience?
Hi Mary,
I fully agree with John Best's remark that "technical advantages of LaB6 aside, the extended life time is worth it". This becomes particularly important if you are going to use automated procedures. We upgraded our JEOL 840 microscope from W to LaB6 filaments because we are often running automated overnight measurements of electron back scattering patterns. With an average life time of 50 hrs for a W filament, there is a probability of 1/3 that an overnight run will be interrupted by a filament burn-out!
Yours sincerely, Joergen. J. B. Bilde-Soerensen Materials Department Risoe National Laboratory DK-4000 Roskilde Denmark
I hope I'm sending this to the right place. This is the first time I have tried to post, and not just reply.
I need some help tracking down a commercial supplier of biotinylated hyaluronic acid binding protien and/or some comments on it's specificity. I am looking for an alternative method of detecting hyaluronic acid in tissue other than enzyme digestion. I have seen HABP used for this, but I read in one article that there is some cross over with chondroitin sulfate, just as there is with most enzymes. Is this true, or is there more than one version of HABP out there?
You might want to try DAB Enhancer from Innovex. It works great Doesn't contain heavy metels and is easy to use. Simply apply for 5 minutes after the DAB step and counter stain. I don't know who the UK supplier is but the US fax # is 510-222-7803. regards, bob
Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis University of North Carolina CB#7400, Rosenau Hall Chapel Hill, NC 27599-7400 Ph. 919-966-6343 Fax 919-966-6123
Hmmm..... This isn't strictly Microscopy, but it is related to using the computer for imaging which is part of our charter, so I guess it's okay........ ;-)
The Apple one is a disaster. Don't buy it. The one I bought (their first & I don't know if there is a second version) has the numeric keypad disconnected. As a result of this disconnection the entire keyboard "system" now has an effective length of nearly1.5 times normal.
This means that if you use the keyboard, keypad and mouse all the time (which I do) then you hand is constantly lifting off the main keyboard and streching to reach the mouse, which is now wayyyyyyyy off to one side. I tried it for a day and then returned it. I started to get a sore shoulder and arm in addition to numb hands.I just use a keyboard at the correct height with a wrist rest.
The only good side is that with my two numb hands I don't paint the walls or rake the grass anymore!
While we are on the subject of TEM screens, has anyone noticed a change in the quality of the screens that are being coated in the recent years? I have always wondered whether the industry has changed their production method or my eyes have just gradually deteriorated over the years.
I have been sending my old screens out for recoating periodically. I have not been totally happy with the screens. Another possibility is that my microscope may be the culprit that it may be contaminating the screen. (Although, recently, I was not happy even with the brand new screens which were just put in the microscope without having too much time to be contaminated.) Susan J.-H. Tao Cheng NIH, Bldg 36, Rm 2A-21 Bethesda, MD 20892 Tel: (301) 496 0579 Fax: (301) 402 6875
} I was told that LaB6 doesn't buy much of a brightness improvement } over W and therefore isn't worth the added expense, is this your } experience?
Hi Mary, I fully agree with John Best's remark that "technical advantages of LaB6 aside, the extended life time is worth it". This becomes particularly important if you are going to use automated procedures running overnight. We upgraded our JEOL 840 microscope from W to LaB6 filaments because we are often running automated overnight measurements of electron back scattering patterns. With an average life time of 50 hrs for a W filament, there is a probability of 1/3 that an overnight run will be interrupted by a filament burn-out !
Yours sincerely, Joergen ------------------------------------------------------------------ J. B. Bilde-Soerensen Materials Department Risoe National Laboratory DK-4000 Roskilde Denmark
Fight this "service contract" tooth and nail, at least for your electron microscopes. These types of services work well for normal routine stuff, but not at all for major repairs. Like any "insurance" they will try and get out of paying for repairs. An example, our SEM developed a vacuum leak, we called our service contract provided, within a day they were here and "fixed it". A week later another vacuum leak occured, this time they came with a mass spec detector and really found the leak. This second time, all day was spent in the lab fixing this "minor" vacuum leak. How would this company your institution is planning on dealing with handle this??
Anyway, fight this! Best of Luck, Ed Calomeni Medical College of Ohio Toledo, OH 43699 emlab-at-opus.mco.edu
I am attempting to evaluate EDS systems for use on an SEM. Has anyone done such an evaluation, and is there documentation you could e-mail me? My samples are typically pharmaceutical powders, packaging material, filter residues, etc. The SEM I use is actually an ElectroScan ESEM, model E-3.
Microscopists, Does anyone have a good protocol for a Sudan Black or other myelin stain that can be used on brain sections? Thanks a bundle. Glen -- Glen T. Prusky Ph.D., Department of Psychology, The University of Lethbridge, 4401 University Drive, Lethbridge, AB, Canada T1K 3M4 pruskyg-at-hg.uleth.ca http://www.uleth.ca/psy/index.htm Office-403-329-5161 Lab-403-329-2410 Fax-403-329-2555
} I am curious as to what other labs use in their TEM closed circuit cooling } systems as an anti-algae, anti-corrosion agent. Are these treatments } successful and how often do they need replacing?
Allan
Once a system is clean, we use about 5-8% ethylene glycol (cheapest= laboratory grade) in distilled water. In systems we run constantly we can= change it every two years if it looks clean. A very slight green tinge= after that time suggests there has been some copper reaction, but it= doesn't worry us. In systems that are not run for a few months, we see= some cloudiness develop.
-- David Rothbard Institute of Paper Science and Technology
The Pacific Northwest Electron Microscopy Society is planning a workshop entitled "Current Techniques in Microtomy of Materials and Life Sciences" presented by Michele Wilhite of RMC. The workshop will be held at the University of Idaho in Moscow, ID on Saturday, June 22, and will include ultramicrotomy of polymers at room temperature and cryo tempertures. Also on the agenda will be cryo-ultramicrotomy and immunolabeling of biological samples, as well as surface preparation for optimum edx and backscatter analysis.
On the evening of the 21st, a dinner buffet and business meeting is planned.
If you are interested in attending either/or, please RSVP by 21 May to jfb-at-uidaho.edu, or fax to (208)885-8937.
While we are on the subject of TEM screens, has anyone noticed a change in the quality of the screens that are being coated in the recent years? I have always wondered whether the industry has changed their production method or my eyes have just gradually deteriorated over the years.
I have been sending my old screens out for recoating periodically. I have not been totally happy with the screens. Another possibility is that my microscope may be the culprit that it may be contaminating the screen. (Although, recently, I was not happy even with the brand new screens which were just put in the microscope without having too much time to be contaminated.)
Susan J.-H. Tao Cheng NIH, Bldg 36, Rm 2A-21 Bethesda, MD 20892 Tel: (301) 496 0579 Fax: (301) 402 6875
The Royal Microscopical Society would like to subscribe to the Microscopy Listserver, and would like to post the following message.
MICRO 96 - International Microscopy Conference and Exhibition
Main theme: Probes in Light, Electron and Digital Microscopy
2-4 July 1996
Novotel, Hammersmith, London W6
Conference topics include:
Green fluorescent proteins Confocal microscopy and 3D imaging Scanning probe microscopy Fluorescent, fluorogenic and luminescent probes Flow cytometry NO synthase Immunogold probes for LM and EM Catalysis Particles and pollution Low energy filtering microscopy Electron backscattering SEM development Biomaterials Paper
Exhibition
There will be an extensive exhibition by 75 companies. For free tickets, a list of exhibitors, stand plan and an exhibition preview please contact the Royal Microscopical Society.
For further details on the MICRO 96 Conference and/or Exhibition or any other RMS activities please contact:
The Royal Microscopical Society 37-38 St Clements Oxford OX1 4AJ UK Tel +44 (0)1865 248768 Fax +44 (0)1865 791237 rms-at-vax.ox.ac.uk
What about the wonderful sounding "tru-form w/pointer" from Adesso? This thing has a two-button touch-pad built into the board at the juncture of the two key pads and it can be used "without removing your hands from the keyboard". I'm really thinking about buying this for exactly the reasons stated below!
} Hmmm..... This isn't strictly Microscopy, but it is related } to using the computer for imaging which is part of our } charter, so I guess it's okay........ ;-) } } The Apple one is a disaster. Don't buy it. The one I bought } (their first & I don't know if there is a second version) } has the numeric keypad disconnected. As a result of this } disconnection the entire keyboard "system" now has an effective } length of nearly1.5 times normal. } } This means that if you use the keyboard, keypad and mouse } all the time (which I do) then you hand is constantly lifting } off the main keyboard and streching to reach the mouse, which is } now wayyyyyyyy off to one side. I tried it for a day and then } returned it. I started to get a sore shoulder and arm in addition } to numb hands.I just use a keyboard at the correct height with a wrist rest. } } } The only good side is that with my two numb hands I don't paint the } walls or rake the grass anymore! } } Nestor } Your Friendly Neighborhood SysOp...... microscopy-at-sparc5.microscopy.commicroscopy-at-sparc5.microscopy.com
Message-Id: {2.2.32.19960517181320.006b5044-at-darkwing.uoregon.edu} X-Sender: mshaf-at-darkwing.uoregon.edu X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
We have an interest to dampen the effects seen due to the electron beam locally heating a silicate (rock) thinsection. The user wants to cool the specimen prior to chamber access, whereas I'm afraid of condensation affecting my ion pumped LaB6 gun. Can anyone alleviate my fears ... or has anyone a remedy??
TIA & cheers, shaf {\/} /\ {\/} /\ {\/} /\ {\/} cognito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/epmahome/
I am looking for sources of used equipment such as equipment brokers etc. Specifically the types of equipment include coating, microscopy and mechanical test equipment. Secondly I'm also trying to locate sources of discounted consumable supplies for metallurgical polishing work.
We have a dual Haskris water chiller, closed recirculating system, that cools our two EM's. Approximately quarterly, I sprinkle a fungicide powder called dichlorophene on top of the water in the reservoir tank in the cooling unit. It floats on top but slowwly disolves over time. We have no problems with stuff growing in the water. We very rarely change the water completely, just add water to the tank once and awhile.
I last purchased dichlorophene powder in 1985 from: K. & K. Labs, 121 Express St., Engineers Hill, Plainview, NY 11803
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"MICROSCOPY & MICROANALYSIS 96" in Minneapolis, Minnesota, Aug.11-15, 1996
Dear Allan, } } We have been recommended to use a product called "Thermoclean DC" as an } anti-algae, anti-corrosion agent in the water of our TEM closed circuit } cooling system. However the data supplied with the product recommends that } the water in the system be replaced every 6-8 months. I personally do not } feel like changing the total 240 litres of water in our two systems this } frequently. } I am curious as to what other labs use in their TEM closed circuit cooling } systems as an anti-algae, anti-corrosion agent. Are these treatments } successful and how often do they need replacing? } We use a product called Aqua Treet 42, which is molybdenum-based (Z = 42 ;-)). We get it from Aqua Laboratories, Inc., P.O. Box 645, 8 Industrial Way, Amesbury MA, USA, (508) 388-3989. We adjust the concen- tration using a test kit obtained from the same company, then adjust the pH with NaOH to 8.0-8.5. We have never had any problems with this since we switched over from a silica-based product (also OK, but no longer avail- able). The water does not have to be changed. We also installed filters in the lines, and this has saved us a lot of grief. These do have to be changed every few months, and we check them monthly along with the concentration of Mo and the pH. We float a little dichlorophene on top of the water in the Haskris circulator to stop the bugs from growing. We get that from K & K division of ICN. Yours, Bill Tivol
This is in response to Greg Erdos' message regarding CIC Agency.
I was "encouraged" last year by our college's administrators to put our 2 scopes on contract w\ CIC. I am approaching the 8 month period now since I did include our new SEM on the contract. The Veterinary teaching hospital here, as well as some other labs are also using this company. Here are some of our observations/experiences.
CIC Agency is a Risk Management Corporation...essentially an insurance company. Instead of contracting with your regular vendor for preventative maintenance, ie JEOL, Phillips, etc., you contract with CIC. Their charge for the service is typically 20-30% below your vendor (its 26% savings on our SEM over JEOL). They include 2 preventative maintenances per year. When you need service, you call whoever you want (ie JEOL) and schedule emergency service. The service engineer orders the parts, does the work as usual, and hands you a bill for parts, labor, and whatever else they charge for. You send the bill to CIC and they pay it. The only stipulation is that you get approval from them if the bill is expected to be over $5000.
Some of the folks here who are using them for hospital equipment are not too pleased and are talking about switching back. I haven't decided yet, but am sure that I won't be putting my TEM with them this upcoming year.
I would be happy to expound on this a bit more with anyone who is interested.
W. L. Steffens, Ph.D Dept. of Veterinary Pathology College of Veterinary Medicine University of Georgia Athens, GA 30602 STEFFENS.B-at-CALC.VET.UGA.EDU Voice: (706) 542-5536 FAX: (706) 542=5828
Netters, Just a note about antifades. Over the years, I have seen or helped folks compare the relative merits of several of these (DABCO, ppd, ehgo) and the results were different each time. Because different types of preparation were being considered (plastic sections, whole mounts and things in between, various chromomophores) I concluded that the relative merits of antifades are specimen/prep dependent. So, if you *really* want the best, you may need to test your own specimen. For the past several years, we have been using a commercial product, "Vecta shield" from Vector labs (I have no interest in this company or product), which has served our needs perfectly (we mostly do plastic sections with cy-3 as a fluorochrome). Hope this helps, Tobias Baskin
I've had and continue to have problems with my wrists. Our safety officer knew of a keyboard in another office and arranged for me to borrow it to try out. I don't remember the brand name, but it was one that has cupped depressions (a strange contraption when you first see it) that accommodate the differences in finger lengths as well as allow the wrists to be in a more natural position. I tried it out for a week. It takes a lot of getting used to because you no longer have to reach for keys as much as on a standard keyboard. I think had I used it for any length of time, it would have been difficult to switch back. And since I use at least 7 different computers at work and home, I didn't think it was worth it. One of these days I want to try the type that just splits the keyboard near the middle and angles the two sides. In my case, that might be the best alternative, although I think that it would still be hard to go back and forth. Perhaps someday the standard keyboard will be adjustable so that you can sit down at any machine and adjust the keyboard (mouse to, maybe) to fit your own hands, much the same way as you adjust the eyepieces on a microscope for you own eyes.
My suggestion is that you try them before you buy them. Check egonomic equipment catalogs and not just computer supply houses.
I, too, have benefited by using something to elevate my wrist once the orignal flareup was under control and my case seems to be mild compared to some peoples.
Regards,
--------------------------------------------------------------------- Jeff L. Brown | Electronics Engineer WL/AADP BLDG 620 | Heterojunction Physics Branch 2241 Avionics Circle RM C2G69 | Electron Devices Division Wright-Patterson AFB OH 45433-7322 | Avionics Directorate | Wright Laboratory (USAF) =====================================================================
Our variety of instruments with their variety of operators have cooling systems maintained in a variety of ways! I don't think we've decided which is best. We have service contracts on our major instruments, so we rely on the contract provider to bail us out if something goes wrong, but anyway... what we mostly use(add) is:
Dichlorophene(2,2-Methylenebis-P-chlorophenol) (Panacide). This is a powder originally suggested by Philips for our TEM purchased circa 1979. We run a Muriatic acid descaler through every few years during an anual maintenace visit.
or... Ethylene Glycol. This is a "conventional wisdom" solution suggested by our air conditioning/heating people, and is what was suggested to way back in my electron microscopy school days. Instruments being fed this stuff also get an occaisional descaling to clean out any corrosion, or scale from hard water. We avoid dH2O due to it's somewhat corrossive nature.
We have been recommended to use a product called "Thermoclean DC" as an anti-algae, anti-corrosion agent in the water of our TEM closed circuit cooling system. However the data supplied with the product recommends that the water in the system be replaced every 6-8 months. I personally do not feel like changing the total 240 litres of water in our two systems this frequently. I am curious as to what other labs use in their TEM closed circuit cooling systems as an anti-algae, anti-corrosion agent. Are these treatments successful and how often do they need replacing?
Thanks in advance.
Allan Mitchell South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
You can get some awfully good results from scanning an image at anything over 100dpi and reprinting with a d2t2 (diesub) printer.
We use the Kodak XLS8600, which gives excellent results in B&W or colour.
############################################################### # # # Don Steele STEELE-at-KRDC.INT.ALCAN.CA # # ALCAN INTERNATIONAL # # Kingston Research and Development Center # # P. O. Box 8400 # # Kingston, Ontario Canada K7L 5L9 # # # ###############################################################
Does anyone out there know how to make high-quality overheads from standard B&W photographs (e.g., TEM images including HRTEM images)? I have always made slides for my talks up until now. Just using the standard xerox transparencies does not give very good results, but I have seen talks where people use transparencies and they are remarkably good. Is there some trick or method I am missing out on?
Roy Christoffersen Texas Center for Superconductivity 3201 Cullen Houston, TX 77204-5932 roy-at-bayou.uh.edu (713) 743-8273 FAX: (713) 743-2787
I am looking into the purchase of a new conventional (W/LaB6) SEM for a Materials Science laboratory and had a couple of questions for users of newer SEM equipment: 1. I was told that LaB6 doesn't buy much of a brightness improvement over W and therefore isn't worth the added expense, is this your experience? 2. With digital imaging as the fast increasing method for documentation, is the record/film option still necessary?
All inputs are greatly appreciated as well as any other comments along these lines. Thanks in advance.
One of the users of our EM Unit wishes to immunogold-label cytoskeletal proteins in cultured fibroblasts. The cells will be lightly fixed, processed by the "progressive lowering of temperature" technique and embedded in Lowicryl K11M. We will use a Leica AFS substitution chamber to process the samples. Unfortnately (for us) this person wishes to keep the cells as a monolayer and section them "face on". We have done this with conventional epoxy resin in the past, however to do this in the AFS chamber may be a little difficult. Has anybody got any suggestions on the substrates to use etc,that would be suitable for PLT processing and embedding in Lowicryl K11M?
Many thanks,
Allan Mitchell South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
We have been recommended to use a product called "Thermoclean DC" as an anti-algae, anti-corrosion agent in the water of our TEM closed circuit cooling system. However the data supplied with the product recommends that the water in the system be replaced every 6-8 months. I personally do not feel like changing the total 240 litres of water in our two systems this frequently. I am curious as to what other labs use in their TEM closed circuit cooling systems as an anti-algae, anti-corrosion agent. Are these treatments successful and how often do they need replacing?
Thanks in advance.
Allan Mitchell South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
} I'm back again! Thanks for all the previous help. I love this } place!! } } Today's problem involves a way to increase gold staining. I have } tissues freshly fixed in 2% PFA (also long term fixed in PLP) and run } up in both LRWhite and Lowicryl K4M. Can these resins be etched to } reveal more structure? Does the antibody and gold label get much } below the surface anyway? } I have increased time in primary to 24hrs. and don't get much } staining. The P.I. on this project has great LM under fluorescence } so we know that the antibody works. I am working with a kit from E.Y. } Laboratories for the other solutions. Any and all help is deeply } appreciated. } } Linda Fox -at-wpo.it.luc.edu
GR Newman and JA Hobot: Modern acrylics for postembedding immunostaining techniques. J Histochem Cytochem 1987; 35:971-981... talks about penetration of reagents into LR White. They found immunoperoxidase got in, but immunogold (they used 10-12nm) stayed on the surface. One quote "Even storage granules within a few nanometers of the section's surface failed to immunolabel".
Personally I hate LR White; nothing ever seems to label! Sorry I can't help with the rest of the problem.
Diana van Driel Dept Ophthalmology Sydney University AUSTRALIA 2006
5 th BRAZILIAN CONFERENCE ON MICROSCOPY OF MATERIALS***MICROMAT 96***
Rio de Janeiro, Brazil October 13-16, 1996
Here is an extract from the first announcement:
Scope of the Conference: MICROMAT 96 follows four previous meetings on Microscopy of Materials [S.Paulo (1988, 1990), Rio de Janeiro (1992), S.Carlos (1994)] sponsored by the Brazilian Society for Electron Microscopy, and will be held at The Marina Palace Hotel in Rio de Janeiro from October 13 to 16, 1996.
Scientific Program: - Electron Microanalysis and Diffraction Techniques - Instrumentation - New Microscopies and Techniques - Application to Materials Research in the Academic and Industrial Context
(Invited lecturers, oral presentations and poster sessions)
Languages: the official languages will be English and Portuguese. No simultaneous translation will be provided.
Conference Proceedings: all communications will be published in a special volume of Acta Microscopica.
Exhibition; Located in the poster zone, a limited number of stands will be available for microscope manufacturers and representatives of microscopy related techniques. Commercial exhibitors should contact conference secretariat for stand reservation and advertisement in the proceedings book.
Conference site and accommodation: The conference hotel is located by the beach with a splendid view of Ipanema and Leblon. October average temperature 25-30 C. A limited number of double occupancy rooms (US$ 40.00 per person) available at the Marina Palace Hotel, other hotels, restaurants within walking distance.
Registration fees: until July 20 th. - BSEM members professional $5o, student $25; non members $90, $50. After July 20, $60, $35, $100, $60.
Post Conference course: minicourse on Digital Microscopy (Cosponsored by BSEM and Gatan Inc.(limited participants!) on Oct.17 and 18, 1996 at The Catholic University of Rio de Janeiro (near the Marina Palace Hotel).
Second circular for those who contact the secretariat will be sent May/June with guidelines for submission of papers and other info.
Chairman:Guillermo Solarzano (PUC-Rio); General Coordinator Luiz Henrique de Almeida (UFRJ) Sponsors: FINEP, CNPQ, FAPERJ, FAPESP, FAPEMIG, FAPERS (all government non-profit agencies)
Contact Secretariat
MICROMAT 96 POBox 38090 22452-970 Rio de Janeiro Brazil Fax + 55 21 5112182 Email sbme-at-rdc.puc-rio.br
Prof.Walter A.Mannheimer Metallurgy and Materials Engineering Federal University of Rio de Janeiro POBox 68505 21945 Rio de Janeiro, Brazil Voice 5521 280-7443 (Dept.office) 5521 590-0579 (direct) Fax 5521 290-6626 Email: wamann-at-metalmat.ufrj.br
Group - David McClellan recently asked for advice on the subject of selection of EDS systems. Due to my rather unique position relating to this thread, I would like to address my response to the full group. As it happens, prior to my current life, I enjoyed reasonably responsibe positions with two of the several major EDS suppliers for some 10 plus years. First, the EDS business is very "mature". When it comes to (pure) EDS hardware and software from the leading 4/5 manufacturers, there is very little difference. Excluded from this generalization is the topic of detectors. Sure, one system might have a very minor "plus" or an equally minor "minus". I expect that a number of readers will not agree with me - most of which, I expect, have become "swift" with one system and happen to find others not so "user friendly" or, otherwise, lacking.. Bottom line, as to the selection of an EDS supplier, I would recommed the following: 1) Find users in YOUR AREA and ask for comments regarding after-sales support. I would not put a great deal of weight on a list that might be supplied by the manufacturer, but rather look for your own inputs. This, I submit, today is the key to your selection! 2) If you have selected your EM manufacturer, you might talk to their applications folks. In doing so, you should understand that they could have their own preferences (based upon their own system knowledge) and that they are conditioned not to recommend one supplier over another. But - if you push, you might learn a bit. David, GOOD LUCK! Don Grimes, Microscopy Today
The Central States Microscopy Society meeting date has been changed to June 28, 1996 due to a conflict occurring during the previous week. Seems that the Street Machines National will be meeting nearby and ALL rooms for a radius of 60 miles have been booked already. Please amend your schedules. I apologize for this and thank Mort Harloe who, being the early-bird that he is, found out about the lack of rooms.
The Spring meeting of the Central States Microscopy Society will take place on June 28, 1996 in Carbondale, IL at the Giant City State Park Lodge. The theme of the meeting will be "Ride the Wave" (e.g., technological wave). Innovative uses of new and established technologies (microwave, LM, SEM, TEM, scanned probes, etc) are invited. Biological and physical science presentations are welcome.
Cash prizes for best student presentations. Typically this ranges from $50-100, depending upon number of entrants and quality of work presented. Abstracts must be received 3 weeks prior to meeting to be considered in the student competition. Follow MSA abstract guidelines.
CSMS corporate sponsors are invited to participate by means of talks and demonstrations of equipment. Contact me well in advance of the meeting, however, if you require tables, space or special electrical setups. There will be no charge for CSMS corporate sponsors.
Programs, lodging recommendations, directions to meeting site will be sent several weeks in advance of the meeting or you may e-mail me for this information.
Speakers and presentors: please contact John Bozzola for topics/timing no later than May 28, 1996.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
To: Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com} *** Reply to note of 05/15/96 16:21
LaB6 ! Yes, it's much better than W. It's about 10x brighter, and lasts much longer. If you can afford it, and not field emission, go for it.
I get 1500 to 2000 hours on a LaB6 tip. Most W filaments last about 40 hours. LaB6 tips cost about $500/each, and it takes the same time to install as W. Alignment time depends on the instrument (ours is a Cambridge S250). W is a mess to clean up, and LaB6 deposits mainly flake off.
Greg, Ed, I couldn't agree more. EM's are special beasts. The EM company's own service is your best bet: calamities WILL HAPPEN (Murphy's law), and your in house service won't want to shell out for a $10K part! Besides, if they're not experienced on working on these complicated pieces of equipment, it may take them forever to figure it out. We have in house service on small stuff (small centrifuges, pH meters, etc.), but all EMs are on EM company contracts.
Sara Miller, Director Diagnostic Virology EM Lab Cancer Center IEM/EM Shared Resource Surgical Pathology EM Lab
On Fri, 17 May 1996, EmLab wrote:
} Date: Fri, 17 May 1996 10:52:56 -0500 (EST) } From: EmLab {EMLAB-at-OPUS.MCO.EDU} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Service contracts } } Dear Greg, } } Fight this "service contract" tooth and nail, at least for your electron } microscopes. These types of services work well for normal routine stuff, but } not at all for major repairs. Like any "insurance" they will try and get out } of paying for repairs. An example, our SEM developed a vacuum leak, we called } our service contract provided, within a day they were here and "fixed it". A } week later another vacuum leak occured, this time they came with a mass spec } detector and really found the leak. This second time, all day was spent in } the lab fixing this "minor" vacuum leak. How would this company your } institution is planning on dealing with handle this?? } } Anyway, fight this! } Best of Luck, } Ed Calomeni } Medical College of Ohio } Toledo, OH 43699 } emlab-at-opus.mco.edu }
============================================================================== Steve Schwarz sschwarz-at-morgan.ucs.mun.ca Dept. of Earth Sciences Memorial University of Newfoundland Newfoundland CANADA A1B 3X5 1-709-737-8142 -737-2589 FAX ******************************************************************************
Message-Id: {9605191055.AA17336-at-super.mhv.net} To: nestor {Microscopy-at-Sparc5.Microscopy.Com}
Good day group,
I would like to partially agree with Don Grimes recommendataions to David Mclellan regarding EDS selection. I thought the marketplace was mature about 5 yeard ago (more or less), when everyone was still using PDP-11's.
I'll suggest that PGT's digital front end and the new spin off companies that are producing inexpensive PC based EDS systems have, are, and will be upsetting the applecart.
First the spin-offs: I think these guys (like EVEX) are doing a great service. The traditional EDS people have been pretty slow in switching over the standard EDS algorithms to low cost PC based platforms. The price of a modern EDS system is roughly half (feature for feature) of a new system five years ago.
The thing that's a bit disturbing is that we're losing some neat tools in the process. Most of the old big 5 (Edax, Kevex-} Fisons, Tracor-} Noran, PGT and Link) had some optional software packages for things like automatically characterizing features based on morphology and chemistry. These days we try do do this using PC based image analysis software on captured images. It's just not as productive as some of the on-the-fly approaches that had beed developed.
I think the most interesting think in EDS these days is a DSP at the front end. I havn't been able to follow up on this, but theoretically, a DSP should be able to deconvolute peaks that formerly had to be rejected because they contained information about two (or more) separate xrays. The improved throughput might make "darn real time xray imaging" a reality. Digital front ends might also make detector sensitivity less of a factor for many applications. Instead of deconvoluting an Xray spectra for elemental IDing, and so called "quantitative" analysis, the digital front end can deconvolute each individual peak, thus producing a truly digital spectrum. Even though DSP's have been in use in other industries for years, I think their impact might ultimately be a huge boon to microscopists.
It would be nice (to the end users advantage, but not necessarily the manufacturers) if ultimately the entire DSP was embedded in a box on the detector assembly and all data was transmitted to the PC via a STANDARDIZED digital interface.
My reccomendation for David Mclelland would be: Buy a really low cost unit at present, because great things are going to be happening in the future with EDS, and you'll want to save your budget for something like a "digital facelift" after this paradym shift has had a chance to settle.
I can't wait to read the groups thoughts on the EDS system of the future.
Regards and Best Wishes to All, John Best -- ELMDAS Co.
We have not had any additive in cooling water with success for many years. Instead, we thoroughly de-ionize the water in the closed circuit system and keep it in this state by passing all the water through carbon filters. If the water needs replacing then we again run it through a large de-ionization unit. With in around 200imp. gallons (800lt) in our system I would not like to be changing the water too often. This system serves 2 EM's, an XPS DPump and another 3 DP's.
alan.wilson-at-dsto.defence.gov.au
Dr Alan Wilson Senior Research Scientist Ship Structures and Materials Division Aeronautical and Maritime Research Laboratory Defence Science and Technology Organization 506 Lorimer St Fishermens Bend 3207 Victoria Australia ph 61 3 9626 7508, fax 61 3 9626 7816 or 61 3 9626 7087
I have been browsing communications on my e-mail and have found your address. Perhaps you can help. My quey is for information regarding the preparation and staining of transverse sections of heather stems ( heavily lignified ) for examination under an optical microscope. These samples have to be softened for sectioning (boiling?). and stained in order to count their annual rings (?) Any help or pointers of who to ask welcome.
Ron Johnston University Wales College Newport johnsto-at-newport.ac.uk
Message-Id: {9605201214.AA07532-at-super.mhv.net} To: nestor {Microscopy-at-sparc5.microscopy.com}
Hello fellow microscopists,
In the short month or so I've been a member of the newsgroup, I haven't seen anything posted on in situ hybridization of nucleic acids and wonder if anyone has information on :
a. past postings on this subject - how can I access them?
b. short courses/workshops in the US in the next six months - can anyone recommend any?
c LR -White embedded tissue - I want to localize viral nucleic acids in plant tissue - using thin sections from samples previously embedded and used for immunolabeling viral proteins Does anyone have any experience, words of wisdom etc on LR White and in situ habridization?
I deeply appreciate any input you can give me - I'm totally new to this technique - haven't even tried it yet!
Peggy
Peggy Brannigan Electron Microscopy Floral and Nursery Plants Research Unit National Arboretum
Dr. Theresa A. Fassel Sr. Research Associate fassel-at-post.its.mcw.edu Department of Microbiology (414)-456-8410 Medical College of Wisconsin Fax (414)-266-8522 8701 Watertown Plank Road Milwaukee, WI 53226-0509
We have the following new printed circuit boards in stock which we are offering for sale at very reduced prices. These parts are for the 450 model but some of them may also be used in earlier models. As these parts are probably no longer available from PMV, you might want to pick some of these parts up for future use. Contact Greg Becker at RMC, Tucson, AZ (520) 889-7900 or e-mail directly at rmcbtli-at-aol.com.
PMV part # Description ( all printedcircuit boards) 14150 F 303 A 14151 F 303 B 14152 F 303 C 14153 F 303 D 14154 F 303 F 14156 F 303 G
Thank you,
Greg Becker RMC 4400 S. Santa Rita Ave Tucson, AZ 85714 tel. (520) 889-7900 fax:(520) 741-2200
Posted-Date: Mon, 20 May 1996 17:02:06 +0200 Message-Id: {31A0896E.7D0C-at-csb.ki.se} Karolinska-at-ondine.csb.ki.se, Institute-at-ondine.csb.ki.se, Depatment-at-ondine.csb.ki.se, of-at-ondine.csb.ki.se, Bioscience-at-ondine.csb.ki.se, Novum-at-ondine.csb.ki.se, S-14157-at-ondine.csb.ki.se, Huddinge-at-ondine.csb.ki.se, Sweden-at-ondine.csb.ki.se Organization: Karolinska Institute X-Mailer: Mozilla 3.0b3 (Win95; I) Mime-Version: 1.0 To: MSA mailing list {Microscopy-at-Sparc5.Microscopy.Com}
Hello everybody,
we have problems with boiling nitrogen in the workstation of a Gatan cryo holder. Particularly large bubbles form near the specimen making transfer very difficult. Has anybody had that problem and found a solution.
On Thu, 16 May 1996 RNBALDUC-at-ARCRIDE.EDU.AR wrote:
} Dear microscopyst: } Please will you e-mail me protocols or technics for rebuild my old TEM screen } thanks in advance } F. Balducci
I would also be interested in this information.
Thomas Moninger (moninger-at-emiris.iaf.uiowa.edu) Central Microscopy Research Facility 85 EMRB University of Iowa Iowa City, IA 52242 319-335-8142 FAX 319-335-8049
I am interested in elucidating the structure of membrane stacks that are roughly 0.5 microns in diameter and 0.2 microns tall. Does anyone out there have any expertise they could share with me on the topic of electron tomography? I know just enough about it to be dangerous and would like more information.
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
A couple of months ago I asked this net about flatbed scanners for my Macintosh. Now I have money and think I have it narrowed down to a Umax Powerlook II or a Microtek Scanmaker III (both come with a variety of software bundles). Does anyone out there have and opinion as to which one might give the best and longest service? Of course, I have to spend the money by Thursday.
Thanks in advance
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
Hello Jeremy, The OTO reaction is one of the many now classic contributions of Professor Jacob S. Hanker when he was a young investigator at Johns Hopkins. "Osmiophilic reagents: new cytochemical principles for light and electron microscopy." Science Vol. 146:1039-1043, 1964. Hanker also published with Seligman, et al. "Histochemical demonstration of some oxidized macromolecules with thiocarbohydrazide (TCH) or thiosemicarbazide (TSC) and osmium tetroxide." J. Histochem. Cytochem. Vol. 13:629-639, 1965. Hope this helps.
Regards to all, Beverly Giammara Center for Clinical Investigations Medical College of Ohio Toledo, OH 43699 419-381-4996 e-mail: Bgiammara-at-gemini.mco.edu
Like Alan Wilson, we too use de-ionized water in our closed circuit system without any additives. This has been the case since 1984 when our system was built. I have a possibility of connecting up to four microscopes and two vacuum evaporators into the circuit, each with their own pumps. I have a filter for each equipment but no active carbon or alike. Everything has worked very fine. The tubing is made of plastic material and everywhere else stainless steel (acid proof). No copper was allowed, because water dissolves very strongly copper.
Regards,
Jouko
Jouko K. M=E4ki, Ph.D., Laboratory Manager University of Turku, Laboratory of Electron Microscopy Kiinamyllynkatu 10 FIN-20520 TURKU FINLAND Tel: +358 21 333 7318 GSM: +358 40 505 2521 FAX: +358 21 333 7380
We use Kodalith to produce photographic quality overheads. I guess other companies manufacture similar products, designed, I believe, for lithographic processes. It's about 1 to 1.5 stops faster than ordinary photographic paper and processes in exactly the same way, apart from drying (like polaroid negatives, you can put it through a dryer but have to be very careful!). With a bit of masking using overheads you can put on micron markers, frames etc. to make it look good. A lot cheaper & quicker than buying a scanner and high resolution laser printer.
Richard Beanland, GMMTL, Caswell, Towcester, Northants NN12 8EQ, UK
} } We have an interest to dampen the effects seen due to the electron beam } locally heating a silicate (rock) thinsection. The user wants to cool the } specimen prior to chamber access, whereas I'm afraid of condensation } affecting my ion pumped LaB6 gun. Can anyone alleviate my fears ... or has } anyone a remedy?? } Dear Shaf, I have found that low-dose imaging aleviates the effects of both heating and charging. I use a 1.2 MV instrument, so our conditions might not be comparable to yours, but you might wish to try using a very low beam current and LoDose or other extremely sensitive film. As dictated by the no-free-lunch principle, you pay for the sensitivity with bigger grain. However, this is a quick and inexpensive thing to try. Good luck. Yours, Bill Tivol
Message-Id: {2.2.32.19960520141720.006b39fc-at-mail.ims.uconn.edu} X-Sender: semlab-at-mail.ims.uconn.edu X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Hi to all,
Thank you for the many helpful responses to my LaB6/digital imaging questions. I am adding a new wrinkle to my SEM purchasing plans (for a Materials Science service laboratory); namely, the variable pressure SEM--which can be run in the conventional or variable pressure mode. I anticipate the use of the variable pressure option to initially be low--less than 20%. My problem is that I want to run a LaB6 filament in conventional SEM mode and occasionally use the variable pressure mode ( the latter buys me a capability I don't presently have). Would I have to change to a W filament for the occasional variable pressure application? I would appreciate any comments, especially regarding potential problems with these variable pressure type SEMs. Examples of uses in the non-biological fields for the variable pressure mode would also be appreciated.
Regards,
Mary Anton University of Connecticut Storrs, CT USA
I have an increasing need to standardise my EPMA for the analysis of natural gold grains, which are usually Au/Ag alloys in the region of 60% (w/w) to 100% Au. The 1995-1996 NIST Catalogue lists such alloys (SRMs 481 and 482), but they seem to be available only in sets of six compositions, for US$384 per set. I really need only one grain (preferably } 250 um) of either the 80% or the 60% Au, and our organisation (particularly the EPMA facility) doesn't have much money. I would be grateful if someone out there could sell me one such grain.
Also, I can find only two synthetic glasses (SRMs 1872 and 1873), one is Pb-rich, the other Ba-rich. I had the impression that they put out some synthetic glasses suitable for use in the analysis of the usual geological major elements. Am I looking in the wrong place, or the wrong catalogue?
Thanks
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Hello Ronald, I have had some limited experience in cutting woody tissue for histological purposes and the most reliable technique has been to simply saok the paraffin-embedded tissue in cool water for at least 12 hours or more. This should allow you to cut reasonable sections at say 10 - 12 microns. If you need more sections than obtained in the first ribbon(s) then just soak the block again. To stain the tissue just use a standard safranin O - fast green technique or simpler still, a toluidine blue O stain. The precise details can be seen in any reasonable plant histotechnique book e.g. 'Plant Microtechnique" by Donald A. Johansen (1940) "Staining procedures used by the biological stain commission' by G. Clark (1973) Hope this helps.
Regards,
Brett Cockman
---------------------------------------------------------------------------- Brett W. Cockman Technologist in Charge School of Dentistry University of Western Australia Voice: (619-2205834) Fax: (619-2213829) e-mail; bcockman-at-uniwa.uwa.edu.au
There is a potential problem with using ethylene glycol in chillers hooked to electron microscopes, and this could arise if some of the water from the chiller system happened to get inside the intstrument column. I know this is an unlikely event, but it has happened in our labs a couple of times. Ethylene glycol is a hygroscopic material that tends to attract and hold water. It can be difficult to get it out of the cracks and crevices inside an instrument, and so it can give rise to problems with the level of vacuum achieved. The algicides, particularly Chloramine-T, usually are only sparingly soluble in water, and they are used in low concentrations. Therefore, a solution of these materials in distilled water would seem to be less likely to leave objectionable deposites inside the instrument in the event of an accident.. I don't know whether or not ethylene clycol is particularly likely to promote corrosion, and would be interested to know if anyone has any authoritative info on this matter. I can't find any reference to it in any of my three books on corrosion. W. C. Bigelow (bigelow-at-umich.edu)
In article alan.wilson-at-dsto.defence.gov.au (Alan Wilson) writes: } Date: Mon, 20 May 1996 13:41:26 +1000 } From: alan.wilson-at-dsto.defence.gov.au (Alan Wilson) } Subject: Re: TEM cooling system additives
} We have not had any additive in cooling water with success for many years. } Instead, we thoroughly de-ionize the water in the closed circuit system and } keep it in this state by passing all the water through carbon filters. If } the water needs replacing then we again run it through a large } de-ionization unit. With in around 200imp. gallons (800lt) in our system I } would not like to be changing the water too often. This system serves 2 } EM's, an XPS DPump and another 3 DP's.
} alan.wilson-at-dsto.defence.gov.au
Distilled (and I imagine de-ionised) water has a more corrosive effect on copper piping than water with additive. Can't give a reference at this date but this was in the (engineering?) literature back in 1967. So we always use additives for fear of corroding cooling pipes inside lenses.
On the other hand we have a 10 micron water filter in line before each of our microscopes, which catches most of the life in our cycled supply.
I have our favorite in situ hybridization protocol on our Web site. See:
http://cellbio.utmb.edu/childs/childs.htm and link to http://cellbio.utmb.edu/childs/cytochem.htm This will provide links to our favorite cytochemical protocols and in situ hybridization is on the list.
Best wishes! Gwen Childs
****************************** Gwen V. Childs, Ph.D. Professor and Vice-Chair Program Director Cell Biology University of Texas Medical Branch Galveston, TX 77555-1043 childs-at-mbian.utmb.edu http://cellbio.utmb.edu/childs/childs.htm
----------
Hello fellow microscopists,
In the short month or so I've been a member of the newsgroup, I haven't seen anything posted on in situ hybridization of nucleic acids and wonder if anyone has information on :
a. past postings on this subject - how can I access them?
b. short courses/workshops in the US in the next six months - can anyone recommend any?
c LR -White embedded tissue - I want to localize viral nucleic acids in plant tissue - using thin sections from samples previously embedded and used for immunolabeling viral proteins Does anyone have any experience, words of wisdom etc on LR White and in situ habridization?
I deeply appreciate any input you can give me - I'm totally new to this technique - haven't even tried it yet!
Peggy
Peggy Brannigan Electron Microscopy Floral and Nursery Plants Research Unit National Arboretum
Yes Rick, I accidentally used the term peak instead of pulse. My dyslexia must be acting up.
You write, "The second use of "peak" and "deconvolution" is confused. It is still not possible at the level of individual photon events in the detector to tell whether a particular photon came from an element line or Bremsstrahlung. This can only be done in a statistical sense looking at the entire spectrum, as has been done for many years. A "digital front end" has no effect on this part of EDS spectrum processing, which has always been digital (as in manipulating numbers in a computer)."
No, I'm not confused. I meant deconvolution of a pulse. Yes, the Bremsstrahlung are a problem in the application of DSP technology to EDS. And finally, of course the Xray spectra is "digital" in the sense that it is an array of numbers manipulated by a computer. But that somehow doesn't do justice to the possibility that a digital front end (Brehmsstralung aside) could eliminate a bunch of a costly analog signal processing hardware by digitizing the pulse shape, not just it's maximum amplitude. Thus the data (from which the familiar spectra is derived) is via digital deconvolution of the pulses and not analog preprocessing of the pulses. Perhaps instead of using the somewhat loose term "purely digital" I should have used something like "digital spectral representation of digitally derived data". (Double digital :) yuk yuk.)
In closing, I'm sure your aware that we're communicating on a relatively low bandwidth medium, and semantic mistakes will sometimes occur. There's no need to be defensive about PGT's technologies. I think what PGT is doing is marvelous for the EM community. If you chose to respond, please do so without malice.
Allan Mitchell asked about controlling algal growth in closed circuit cooling systems. This can be a very real problem for most electron microscopes. There are three things that can be done to control it; 1. Algal growth can be greatly retarded by excluding light from all parts of the cooling system. This involves using opaque tubing, and keeping the water reservoir in the water chiller covered with a light-tight cover at all times. 2. Further control can be achieved by using an algicide. The old standby is a product known as Chloramine-T, which is the sodium salt of N-chloro-p-toluenesulphonamide. This is available from most specialty chemical companies (e.g. Polysciences, Sigma, Aldrich), and is used at the level of about 1 gram per gallon of water in the cooling system (0.25 g/litre). 3. A filter should be installed on the intake to the water line to prevent algae and other solid materials from getting into the cooling lines of the lenses, etc. This filter must be cleaned, and preferably replaced, on a regular basis. ( Ref: Vacuum Methods in Electron Microscopy, by W. C. Bigelow, Portland Press, 1994, p. 216.) I have not had any experience with the product called Thermoclean DC; however, we have used the Chloramine-T stuff in our several systems here at the Univ. of Michigan for a number of years with very good success. We use distilled water to avoid scale formation in heated parts of the system (the stuff sold in drugstores is good enough), and add more as necessary from time to time to keep the system up to the necessary operating level. We only replace it when it gets dirty or otherwise contaminated, or when it is lost due to service problems. Good luck with your system Wil Bigelow (bigelow-at-umich.edu)
X-Mailer: WinNET Mail, v2.30 Message-ID: {946-at-oimag.win-uk.net} Reply-To: Software department {software-at-oimag.win-uk.net} To: rick-at-pgt.com, Microscopy-at-Sparc5.Microscopy.Com
Thank you Rick for clarifying the confusion between deconvolution of spectral peaks and separating individual signal pulses arising from the x-ray detector. I certainly echo the comments in your last paragraph.
I feel the words "The first sense, in which pulse pile-up can be untangled, is no longer in the realm of theory." are rather overstating the case. Practical circuits to detect pulse pile-up have been available and present in equipment for over 20 years so this issue is hardly "in the realm of theory". The major advances in recent years have been in improved electronic detection of low energy x-ray events ( { 1 keV ) which reduces pile and gives better spectral fidelity in this low energy region.
Now that high speed ADC's are available at reasonable cost, a digital electronic implementation is considerably more convenient and flexible than playing around with delay lines and complex time-variant analogue circuitry. Nevertheless, all electronic processors are faced with the same problem: the major component of noise at the detector head is inextricably linked with the photon signal before the signal is digitised. "Adaptive pulse shaping" has both benefits and disadvantages depending on the intended application but as far as I am aware, it cannot tell the electrons in the detector and FET when and where to move.
I am also one of those commercial guys who wants everyone to buy an EDX system... I hope for all the right reasons.
Peter Statham Oxford Instruments Microanalysis Group
} John Best writes: } } {... skipping some interesting opinions ...} } } } I think the most interesting think in EDS these days is a DSP at the } } front end. I havn't been able to follow up on this, but theoretically, a } } DSP should be able to deconvolute peaks that formerly had to be rejected } } because they contained information about two (or more) separate xrays. } } The improved throughput might make "darn real time xray imaging" a } } reality. Digital front ends might also make detector sensitivity less of } } a factor for many applications. Instead of deconvoluting an Xray spectra } } for elemental IDing, and so called "quantitative" analysis, the digital } } front end can deconvolute each individual peak, thus producing a } } truly digital spectrum. } } The word "peak" is being used for two things here. The first sense, in } which pulse pile-up can be untangled, is no longer in the realm of theory. } This is what PGT's adaptive pulse processing does, and has been commercially } available since 1993. The word "peak" is being used to describe the time } waveform emerging from the detector preamplifier or any subsequent analog } stages before digitization; perhaps "pulse" is a more usual term. } } The second use of "peak" and "deconvolution" is confused. It is still not } possible at the level of individual photon events in the detector to tell } whether a particular photon came from an element line or Bremsstrahlung. } This can only be done in a statistical sense looking at the entire } spectrum, as has been done for many years. A "digital front end" has } no effect on this part of EDS spectrum processing, which has always been } digital (as in manipulating numbers in a computer). } } DISCLOSURE: I work for PGT, so I have an obvious interest in the outcome } of this debate. } } Regards to all, } } Rick Mott } rick-at-pgt.com }
-- -- Software Dept.( shared email facility ) -- Oxford Instruments Microanalysis Group -- Halifax Road, High Wycombe, Bucks HP12 3SE, UK
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-- [ From: Charles A. Garber., Ph.d * EMC.Ver #2.10P ] --
In response to: ================ While we are on the subject of TEM screens, has anyone noticed a change in the quality of the screens that are being coated in the recent years? I have always wondered whether the industry has changed their production method or my eyes have just gradually deteriorated over the years. ======================================
For year and years, the optimum phosphors used for TEM screens were Cd based, until that is, people found out how really bad of an actor (e.g. highly toxic) these Cd based phosphors really were. As a result, the phosphor industry pretty much reformulated their phosphors with less toxic alternatives, however for the TEM application, I have certainly been led to believe that the "substitutes" are clearly not equivalent by any sense of the word.
From what I have seen with my own eyes over the years, going back to my own graduate school career, at least in my own experience which might not be representative, the coating of screens was never done with the respect deserving of such toxic compounds. I have personally been convinced that this is something better left in the hands of people who do this for a full time living.
SPI has offered a screen recoating service for some years. We do not do this in house, but in an outside facility dedicated to the coating of phosphor screens. I have been told that they have a "life time" supply of the original Cd based phosphor. I do not permit this kind of activity to be performed on our premises because of the safety concerns. We have stopped offering the Cd based phosphors as part of our business for the same liability concerns that have caused the manufacturers to stop producing the original materials.
Another alternative to the need to periodically recoat screens is to replace the center of the screen with a YAG single crystal scintillator disc. While the initial purchase price might seem a bit on the high side, the cost is probably equivalent to two or three screen recoatings. And you do not have to put up with the down time associated with the changing of screens. Nothing in life is forever, of course, but the YAG single crystal disc comes close to it, and I think it would be safe to say that it would be impossible for a "normal" TEM beam to "burn" a hole in one of our SPI single crystal YAG scintillator discs being used as a TEM screen. At least I have not ever been made aware of anyone who has ever burned such a hole in a YAG disc modified screen.
Disclaimer: SPI Supplies both recoats screens and also supplies the YAG single crystal scintillator discs so we would clearly have an interest in customers not coating their own screens!
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: spi2spi-at-2spi.com
} } While we are on the subject of TEM screens, has anyone noticed a change in } the quality of the screens that are being coated in the recent years? I } have always wondered whether the industry has changed their production } method or my eyes have just gradually deteriorated over the years. } } I have been sending my old screens out for recoating periodically. I have } not been totally happy with the screens. Another possibility is that my } microscope may be the culprit that it may be contaminating the screen. } (Although, recently, I was not happy even with the brand new screens which } were just put in the microscope without having too much time to be } contaminated.) } Dear Susan, For many years we had coated our own screens using a method which involved suspending phosphor in gelatin and letting it settle out of sus- pension. We got very good screens when things worked properly, but often there would be inhomogeneities, and the skill in pouring fades when not used often. The gelatin-based screens develop a brownish cast over the course of about a year and fade accordingly. We recently obtained screens from Grant Scientific and Fullam which were prepared my a method using an organic solvent. We have just installed three of the screens from Grant--a high-brightness screen, a focussing screen, which has a central low-brightness-high-resolution insert, and a transmission screen for our video system. We have evaluated these screens vs our old ones, and we find that our old screens are brighter, but the Grant screens are very uniform and of high resolution. The focussing screen is of exceptional quality, and the high-brightness and video screens are pretty good, although we would prefer them to be brighter. I am going to talk to Grant to see if future screens could be made brighter, and I have no reason to believe that they could not. I have no idea as yet whether the Grant screens will have longer or shorter useful lives than our old type, and I have not had a chance to evaluate Fullam's (high-brightness) screen. Yours, Bill Tivol
} Return-Path: Microscopy-request-at-Sparc5.Microscopy.Com } Date: Fri, 17 May 1996 10:27:19 -0500 } From: Linda Fox {lfox1-at-wpo.it.luc.edu} } To: Microscopy-at-aaem.amc.anl.gov } Subject: RE: TEM cooling system additives } } The water in our recirculating unit was recently changed under our } yearly service contract. The engineer filled it with water from our } Milli-Q system and had us order one liter of Ethylene Glycol (Fisher } Scientific) to add to the system. I just checked, and there is } nothing growing, that I can see, however there is a faint metallic } sheen across surface of the water. Hope this helps. } Linda M. Fox lfox1-at-wpo.it.luc.edu } }
Help me! I need to unsubscribe from the bulletin board and I don't know how. Can someone send me some instructions?
psic-at-uclink4.berkeley.edu
Doug Davis Staff Research Associate Electron Microscope Lab University of California Berkeley, CA 94720 (510) 642-2085, fax 643-6207 dbd1-at-uclink4.berkeley.edu
At 05:18 PM 5/16/96 -0600, you wrote: "The 10x brightness which has been touted for years only occurs for smaller cone angles (60 degrees)."
LaB6 cathodes with 60 degree cone angles are readily available for all TEMs and SEMs, although they are more commonly used with TEMs.
I would be happy to supply technical data sheets on LaB6 brightness to anyone who is interested.
Disclaimer: Energy Beam Sciences manufactures tungsten filaments and distributes Denka LaB6 cathodes.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
} Return-Path: Microscopy-request-at-Sparc5.Microscopy.Com } Date: Fri, 17 May 1996 08:49:50 -0500 } To: Microscopy-at-Sparc5.Microscopy.Com } From: David.Rothbard-at-ipst.edu (David Rothbard) } Subject: Re: TEM cooling system additives } } } I am curious as to what other labs use in their TEM closed circuit cooling } } systems as an anti-algae, anti-corrosion agent. Are these treatments } } successful and how often do they need replacing? } } Allan } } Once a system is clean, we use about 5-8% ethylene glycol (cheapest laboratory grade) in distilled water. In systems we run constantly we can change it every two years if it looks clean. A very slight green tinge after that time suggests there has been some copper reaction, but it doesn't worry us. In systems that are not run for a few months, we see some cloudiness develop. } } -- } David Rothbard } Institute of Paper Science and Technology } } }
} I would have thought that totally de-ionized water would cause a } problem, since it is "seeking ions", which can lead to } corrosion. This is the same reason that you never want to } put deionized water in your car's radiator. Perhaps } you mean "distilled water"?
No, the water is initially deionized through a resin exchange system. One point to note is that all of the plumbing is copper with some brass fittings. (and stainless steel in the microscopes). This system has run for } 10 years with no problems. One of our "plumbers" put a galv. fitting in the system which was noticed after a couple of days. The fittings was severely attacked in this short time. Thus a caveat may be required - use deionized water only if the plumbing is single metal, or copper/brass.
alan.wilson-at-dsto.defence.gov.au
Dr Alan Wilson Senior Research Scientist Ship Structures and Materials Division Aeronautical and Maritime Research Laboratory Defence Science and Technology Organization 506 Lorimer St Fishermens Bend 3207 Victoria Australia ph 61 3 9626 7508, fax 61 3 9626 7816 or 61 3 9626 7087
Dear Philip: To help alleviate the bubbling disturbances in the workstation during specimen transfer, we have found that the insertion of a piece of filter paper approximately 2cm x 4cm in a vertical position over the hole where the holder enters the workstation is worthwhile. To reduce the chance of the grid bubbling out of the holder after transfer and before replacement of the hexring, use a liquid nitrogen level just above the holder tip. If you are fast, the split second or so that the grid is in the cold band above the liquid surface will not affect ice quality. Alternatively, try a heavier grid such as molybdenum; it won't bounce around as much. Good luck. We can discuss further through direct E-Mail.
Donald Gantz Boston Univ Med Schoo Gantz-at-Med-biophd.bu.edu
} } -- [ From: Charles A. Garber., Ph.d * EMC.Ver #2.10P ] -- [skip] } Nothing in life is forever, } of course, but the YAG single crystal disc comes close to it, and I } think it would be safe to say that it would be impossible for a } "normal" TEM beam to "burn" a hole in one of our SPI single crystal } YAG scintillator discs being used as a TEM screen. At least I have not } ever been made aware of anyone who has ever burned such a hole in a YAG } disc modified screen.
Dear Chuck, Is this also true for those operating in diffraction mode? I have heard of YAG developing a permanent set of dark spots, which would be most distracting. Yours, Bill Tivol
} Return-Path: Microscopy-request-at-Sparc5.Microscopy.Com } Date: Fri, 17 May 1996 10:52:56 -0500 (EST) } From: EmLab {EMLAB-at-OPUS.MCO.EDU} } Subject: Re: Service contracts } To: microscopy-at-Sparc5.Microscopy.Com } X-VMS-To: IN%"microscopy-at-msa.microscopy.com" } } Dear Greg, } } Fight this "service contract" tooth and nail, at least for your electron } microscopes. These types of services work well for normal routine stuff, but } not at all for major repairs. Like any "insurance" they will try and get out } of paying for repairs. An example, our SEM developed a vacuum leak, we called } our service contract provided, within a day they were here and "fixed it". A } week later another vacuum leak occured, this time they came with a mass spec } detector and really found the leak. This second time, all day was spent in } the lab fixing this "minor" vacuum leak. How would this company your } institution is planning on dealing with handle this?? } } Anyway, fight this! } Best of Luck, } Ed Calomeni } Medical College of Ohio } Toledo, OH 43699 } emlab-at-opus.mco.edu } }
} Return-Path: Microscopy-request-at-Sparc5.Microscopy.Com } Date: Fri, 17 May 1996 14:21:19 -0500 } From: "Gib Ahlstrand" {giba-at-puccini.crl.umn.edu} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Re: TEM cooling system additives } } We have a dual Haskris water chiller, closed recirculating system, that cools } our two EM's. Approximately quarterly, I sprinkle a fungicide powder called } dichlorophene on top of the water in the reservoir tank in the cooling unit. It } floats on top but slowwly disolves over time. We have no problems with stuff } growing in the water. We very rarely change the water completely, just add water } to the tank once and awhile. } } I last purchased dichlorophene powder in 1985 from: K. & K. Labs, 121 Express } St., Engineers Hill, Plainview, NY 11803 } } Gib Ahlstrand, MMS Newsletter Editor } Electron Optical Facility, University of Minnesota, Dept. Plant Pathology } 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 } 612-625-9728 FAX, giba-at-puccini.crl.umn.edu } } "MICROSCOPY & MICROANALYSIS 96" in Minneapolis, Minnesota, Aug.11-15, 1996 } }
A colleague of mine is looking for a tissue chopper (NOT a Vibratome). Does anyone have any info that might help her?
I've heard that the tissue chopper might be back in production after several years' absence, but I haven't been able to get any details. Maybe this is just a rumor??
I would appreciate any information you might be able to provide. Thanks in advance.
Bob Chiovetti Applications Lab Manager RMC 1-800-453-2242 Fax 520-741-2200 RMCBTLI-at-aol.com
} Return-Path: Microscopy-request-at-Sparc5.Microscopy.Com } X-Sender: alan.wilson-at-SoMPop.dsto.defence.gov.au } Date: Mon, 20 May 1996 13:41:26 +1000 } To: Microscopy-at-Sparc5.Microscopy.Com } From: alan.wilson-at-dsto.defence.gov.au (Alan Wilson) } Subject: Re: TEM cooling system additives } } We have not had any additive in cooling water with success for many years. } Instead, we thoroughly de-ionize the water in the closed circuit system and } keep it in this state by passing all the water through carbon filters. If } the water needs replacing then we again run it through a large } de-ionization unit. With in around 200imp. gallons (800lt) in our system I } would not like to be changing the water too often. This system serves 2 } EM's, an XPS DPump and another 3 DP's. } } alan.wilson-at-dsto.defence.gov.au } } Dr Alan Wilson } Senior Research Scientist } Ship Structures and Materials Division } Aeronautical and Maritime Research Laboratory } Defence Science and Technology Organization } 506 Lorimer St } Fishermens Bend 3207 } Victoria Australia } ph 61 3 9626 7508, fax 61 3 9626 7816 or 61 3 9626 7087 } } }
I used to have 2 AEI 801 electron microscopes and always recoated my screens. It can be frustrating at times but I got real nice screens. I used the phoshor from JEOL and I got good contrast and brightness on the screen. I tried other phosphors from different companies but the JEOL phosphor seemed to work the best. If anyone is interested, I can fax a copy of how I recoated the screen. I will e-mail if you'd rather. No I am not connected with JEOL. I do have 2 JEOL scopes and 1 Zeiss scope.
1) Every instrument I used since 1979 (Joel, Zeiss, Hitachi) contained glycol in the water and have yet to detect problems outlined in this discussio:
a) Make sure that you contact the chiller manufacturer to get proper ratio of water to glycol, b) Yes, glycol COULD harden seals with leak potential. I think that regular maintenance and replacement of old glycol can help. c) Mix with the glycol only dianized or distilled water, never tap water!
********************************************************************* *Cesar D. Fermin, Ph.D \|T|/ Fax (504) 587-7389 * *Tulane Medical School /|M|\ Voice Mail (504) 584-2618 * *Pathology/SL79 \|C|/ Secretary (504) 584-2436 * *New Orleans La 70112-2699 /|*|\ Lab/Techn. (504) 584-2521 * * {Fermin-at-tmc.tulane.edu} ----} Prof. Pathology & Otolaryngology * *http://www1.omi.tulane.edu/departments/pathology/fermin/cdftop.html* *********************************************************************
} Return-Path: Microscopy-request-at-Sparc5.Microscopy.Com } Date: Thu, 16 May 1996 11:23:55 GMT } X-Sender: gwe-at-biotech.ufl.edu } To: Microscopy-at-Sparc5.Microscopy.Com } From: gwe-at-biotech.ufl.edu (Greg Erdos) } Subject: Service contracts } } The University of Florida is strongly urging all of us to drop our } service contracts on all types of equipment in favor of going with a company } called CIC Agency who will handle service payments. We chose the service } provider and they pay for the services charges. It sort of like an HMO. We } pay a fixed annual fee. } } Has anyone had experience with CIC or similar? If so could you share with } us your feelings. } ******************************************************* } Greg Erdos Phone: 352-392-1295 } Scientific Director, } ICBR Electron Microscopy Core Lab } 218 Carr Hall Fax: 352-846-0251 } University of Florida E-mail: gwe-at-biotech.ufl.edu } Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ } } ******************************************************* } }
} Return-Path: Microscopy-request-at-Sparc5.Microscopy.Com } Alternate-Recipient: allowed } Auto-Forwarded: prohibited } Content-Return: allowed } Disclose-Recipients: prohibited } Conversion: allowed } Importance: normal } Priority: normal } Sensitivity: Company-Confidential } Subject: RE: TEM cooling system additives } From: "Scott D. Walck WL/MLBT" {walcksd-at-ml.wpafb.af.mil} } To: Linda Fox {lfox1-at-wpo.it.luc.edu} , } Microscopy ListServer {Microscopy-at-Sparc5.Microscopy.Com} } Date: Fri, 17 May 96 20:30:21 -0400 } } } The water in our recirculating unit was recently changed under our } } yearly service contract. The engineer filled it with water from our } } Milli-Q system and had us order one liter of Ethylene Glycol (Fisher } } Scientific) to add to the system. I just checked, and there is } } nothing growing, that I can see, however there is a faint metallic } } sheen across surface of the water. Hope this helps. } } Linda M. Fox lfox1-at-wpo.it.luc.edu } } } You might want to check this out, but I once was told by a reliable source (professor that taught and worked in corrosion) that ethylene glycol by itself is corrosive and that in antifreeze there are antioxidants and such added to it to prevent corrosion. Now I might be suffering from old age and this was given to me a long time ago, but I'm pretty sure that I got this right. If anyone out there in microland knows for sure, I would like to know if what I'm relaying here is in fact true. } } - -Scott Walck } }
We have complimentary 1 day exhibit passes available for the 5th World Biomaterials Congress held in Toronto May28th -June 2nd in Toronto.
We just got a hold of these tickets & I'd like to notify individuals who were planning on attending to see Digital Instrument's SPM at the Congress. These passes will probably save you about $75.
If this applies to you, please contact Art Priebe at 613-832-0094 or E-mail us at sys4rsch-at-travel-net.com.
(We are the Canadian Representatives for D.I.)
Thankyou
Mark Priebe
Mark Priebe Sales Representative Systems for Research Corp (sys4rsch-at-travel-net.com)
In message Mon, 20 May 1996 13:53:57 +0100 (BST), Ronald Johnston {johnston-at-sun1.newport.ac.uk} writes:
} I have been browsing communications on my e-mail and have found your } address. Perhaps you can help. } My quey is for information regarding the preparation and staining of } transverse sections of heather stems ( heavily lignified ) for } examination under an optical microscope. } These samples have to be softened for sectioning (boiling?). } and stained in order to count their annual rings (?) } Any help or pointers of who to ask welcome. } } Ron Johnston } University Wales College Newport } johnsto-at-newport.ac.uk } ----------- If you are basically interested in counting the annual ring, a lignin stain such as Phlolroglucin should give you very quick results. Place the sections in 0.1 gram of phloroglucin in 10 cc of 95% ETOH for a few minutes and transfer to 25% HCL. Lignified tissue will appear red-violet. If your material would lend itself to it, you may try even a faster method by staining the cut end of the wood, provided the cut is smooth. You could "paint" the cut end with phloroglucin and HCL and view the wood under a dissection microscope to count the annual rings! Good Luck!
************************************************************************* M.V. Parthasarathy Professor of Plant Biology & Director, Cornell Integrated Microscopy Center 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 ***********************************************************************
} we have problems with boiling nitrogen in the workstation of a Gatan } cryo holder. Particularly large bubbles form near the specimen making } transfer very difficult. Has anybody had that problem and found a } solution.
Dear Philip, Once everything gets cold, there should no longer be much boiling. If you have the heater running, or if the temp has not reached steady state, there will be problems. I usually pour a lot of LN2 in the station and after it reaches steady state, I pour some more LN2 in so that there is liquid in the depressions in the aluminum piece surrounding the stage, but the level is below that of the specimen. The secret is to be very pa- tient about everything and not to worry if a little LN2 comes out the side of the station (where the stage goes in). Good luck. Yours, Bill Tivol
On Mon, 20 May 1996 wise-at-vaxa.cis.uwosh.edu wrote:
} To all, } } A couple of months ago I asked this net about flatbed scanners for } my Macintosh. Now I have money and think I have it narrowed down to a Umax } Powerlook II or a Microtek Scanmaker III (both come with a variety of } software bundles). Does anyone out there have and opinion as to which one } might give the best and longest service? Of course, I have to spend the } money by Thursday. } } Thanks in advance } } Bob } } } Robert R. Wise, PhD } Director, UWO Electron Microscope Facility } Department of Biology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } (414) 424-3404 tel } (414) 424-1101 fax } wise-at-vaxa.cis.uwosh.edu }
We have had a Microtek IIHR with the transparency adaptor for a few months now and have been quite pleased. It came with full Adobe Photoshop (which we wanted anyway) and some optical character recognition software. We had a bit of a conflict with the 16 bit scanner running under 32 bit '95 but a fix-it DLL from Adobe cured the problem. THe only other problem we have is a bit of inconsistancy in image quality when scanning dense TEM negs, and that can usually be solved by adjusting the setting manually. From what I have heard the III is equal to or better than the IIHR. Hope this helps.
Tom
Thomas Moninger (moninger-at-emiris.iaf.uiowa.edu) Central Microscopy Research Facility 85 EMRB University of Iowa Iowa City, IA 52242 319-335-8142 FAX 319-335-8049
} Return-Path: Microscopy-request-at-Sparc5.Microscopy.Com } From: William Tivol {tivol-at-wadsworth.org} } Subject: Re: TEM cooling system additives } To: richard.easingwood-at-stonebow.otago.ac.nz (Richard Easingwood) } Date: Fri, 17 May 1996 11:21:56 -0400 (EDT) } Cc: microscopy-at-Sparc5.Microscopy.Com } Content-Length: 1463 } } Dear Allan, } } } } We have been recommended to use a product called "Thermoclean DC" as an } } anti-algae, anti-corrosion agent in the water of our TEM closed circuit } } cooling system. However the data supplied with the product recommends that } } the water in the system be replaced every 6-8 months. I personally do not } } feel like changing the total 240 litres of water in our two systems this } } frequently. } } I am curious as to what other labs use in their TEM closed circuit cooling } } systems as an anti-algae, anti-corrosion agent. Are these treatments } } successful and how often do they need replacing? } } } We use a product called Aqua Treet 42, which is molybdenum-based } (Z = 42 ;-)). We get it from Aqua Laboratories, Inc., P.O. Box 645, 8 } Industrial Way, Amesbury MA, USA, (508) 388-3989. We adjust the concen- } tration using a test kit obtained from the same company, then adjust the } pH with NaOH to 8.0-8.5. We have never had any problems with this since } we switched over from a silica-based product (also OK, but no longer avail- } able). The water does not have to be changed. } We also installed filters in the lines, and this has saved us a lot } of grief. These do have to be changed every few months, and we check them } monthly along with the concentration of Mo and the pH. } We float a little dichlorophene on top of the water in the Haskris } circulator to stop the bugs from growing. We get that from K & K division } of ICN. } Yours, } Bill Tivol } }
In message Mon, 20 May 1996 10:25:44 -0400, brannign-at-asrr.arsusda.gov (Peggy Brannigan) writes:
} Hello fellow microscopists, } } In the short month or so I've been a member of the newsgroup, I haven't } seen anything posted on in situ hybridization of nucleic acids and wonder } if anyone has information on : } } a. past postings on this subject - how can I access them? } } b. short courses/workshops in the US in the next six months - can anyone } recommend any? } } c LR -White embedded tissue - I want to localize viral nucleic acids in } plant tissue - using thin sections from samples previously embedded and } used for immunolabeling viral proteins Does anyone have any experience, } words of wisdom etc on LR White and in situ habridization? } } I deeply appreciate any input you can give me - I'm totally new to this } technique - haven't even tried it yet! } } Peggy } } Peggy Brannigan } Electron Microscopy } Floral and Nursery Plants Research Unit } National Arboretum } } Bldg. 010A R.238 } 10300 Baltimore Avenue } Beltsville, MD USA20705 } } Phone: (301) 504-6097 } Fax : (301) 504-5096 } Email: brannign-at-asrr.arsusda. gov } -------------- Although a few have used LR White embedded tissue for in situ hybridization studies, you will find that most prefer Lowicryls. I suggest that you refer to the recent article by Xingxiang Li & Thomas W. Okita (1995), "Localization of RNA by high resolution in situ hybridization" In: Methods in Plant Cell Biology; Edts. D.W. Galbraith, H.J. Bohnert & D.P. Bourque, Vol 49, Part A, pages 185-199; Academic Press. Good Luck!
************************************************************************* M.V. Parthasarathy Professor of Plant Biology & Director, Cornell Integrated Microscopy Center 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 ***********************************************************************
} Return-Path: Microscopy-request-at-Sparc5.Microscopy.Com } From: "Buddy Steffens" {STEFFENS.B-at-calc.vet.uga.edu} } Organization: College of Vet. Med } To: gwe-at-biotech.ufl.edu (Greg Erdos), Microscopy-at-Sparc5.Microscopy.Com } Date: Fri, 17 May 1996 12:48:18 EST } Subject: Re: Service contracts w\ CIC Agency } Priority: normal } } This is in response to Greg Erdos' message regarding CIC Agency. } } I was "encouraged" last year by our college's administrators to put } our 2 scopes on contract w\ CIC. I am approaching the 8 month period } now since I did include our new SEM on the contract. The Veterinary } teaching hospital here, as well as some other labs are also using } this company. Here are some of our observations/experiences. } } CIC Agency is a Risk Management Corporation...essentially an } insurance company. Instead of contracting with your regular vendor } for preventative maintenance, ie JEOL, Phillips, etc., you contract } with CIC. Their charge for the service is typically 20-30% below } your vendor (its 26% savings on our SEM over JEOL). They include 2 } preventative maintenances per year. When you need service, you call } whoever you want (ie JEOL) and schedule emergency service. The } service engineer orders the parts, does the work as usual, and hands } you a bill for parts, labor, and whatever else they charge for. You } send the bill to CIC and they pay it. The only stipulation is that } you get approval from them if the bill is expected to be over $5000. } } Some of the folks here who are using them for hospital equipment are } not too pleased and are talking about switching back. I haven't } decided yet, but am sure that I won't be putting my TEM with them } this upcoming year. } } I would be happy to expound on this a bit more with anyone who is } interested. } } } } } } W. L. Steffens, Ph.D } Dept. of Veterinary Pathology } College of Veterinary Medicine } University of Georgia } Athens, GA 30602 } STEFFENS.B-at-CALC.VET.UGA.EDU } Voice: (706) 542-5536 } FAX: (706) 542=5828 } }
Workshop on Specimen Preparation for Transmission Microscopy in The Physical and Life Sciences
Sponsored by
Midwest Microscopy & Microanalysis Society (MMMS)
June 7 & 8, 1996
____________ Location ____________ Argonne National Laboratory Materials Science Division Building 212 9700 S. Cass Ave Argonne, Illinois 60439
Organizer: Nestor J. Zaluzec
____________ Dates & Times ____________
June 7 - Physical Sciences 10 am - 4 pm
June 8 - Life Sciences 10 am - 4 pm
AM - Plenary Invited Lectures PM - Hands on Workshop/Lab
____________ Registration ____________
All visitors must Pre-Register and obtain a site pass for entry to Argonne National Laboratory. Call Sheila Jungman at (708)-252-4987 or register electronically on the WWW at
MMMS Members - None NonMembers - $10.00 Students - $5.00
Registration Fees are payable at the meeting.
__________________ Meeting Format ___________________
The meeting will consist of morning lectures followed by afternoon hands on workshops. All presentations will be by invited speakers. An exhibit area will be available for Corporate Members.
______________________ Workshop Program ______________________ Physical Sciences ______________________
* Electro-Chemical Polishing B. Kestel - ANL
* Ion Milling, Mechanical Thinning, & Cross-sectioning R. Alani - Gatan Inc.
* Microtomy of Hard Materials P. Swab-Adv. Refract. Tech.
__________________ Life Sciences __________________
* Introd. to Staining and Fixation G. Scott - Univ. of Wisc.
* Immuno-Gold Labeling R. Albrecht- Univ. of Wisc.
* Microwave Fixation Techniques L. Dickey - Micro Lines Mrkt.
* CryoMicrotomy for Immunocytochemical Studies M. Wilhite - Consultant
Thanx for your comments. One question that comes to mind is whether there will be a special charge from the microscope companies to re-enter their service contract program if the cheaper service doesn't work out. It still may be worth it, but the figures are things one should know up front before switching. Sara
On Mon, 20 May 1996 amorr-at-mse.ufl.edu wrote:
} Date: Mon, 20 May 96 13:16:15 EST } From: amorr-at-mse.ufl.edu } To: Sara Miller {saram-at-acpub.duke.edu} , EMLAB-at-OPUS.MCO.EDU } Cc: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Service contracts } } Greg, Sara, Ed: } Changes always give us feelings of uncertainty and thus we } resist them. Greg's initiative will give us insight through } responses from people who used this type of service } contract companies before, and hence will alleviate those } fears. Another source of information are the } "orientation" meetings that these companies are giving to } prospective clients. Although I missed one such meeting, I } heard some interesting comments from someone who } attended. Based on those comments, I would like to } add to Ed's and Sara's response to Greg that we } should be informed on what each company is offering under } the guidelines of the client institution (say, our } university). In this particular case, the company (that I } believe already won the univ. bid) does not propose to have } their own repair engineers do the work, but rather gives us } the choice of whom to call. So, we can still call JEOL to } repair our Jeol EMs, Hitachi to fix our Hitachi instruments, } etc (or, more ineterstingly, the other way around). } Also, if this does not work out, we can discontinue the } service and go back to our present service contracts. A } big point in favor of the new system is that it is cheaper, } and the prices are guaranteed for 2 years. } } I look forward to see more posting on this issue. } } Augusto Morrone } Univ. of Florida } MSE }
It is nice to see a thread on energy-dispersive spectrometry get going here. The original question dealt with selecting an EDS system from the current systems available. There is no question that some really neat advances have been made in the hardware arena for pulse processing etc. I am not really qualified to comment on such things.
However, as a user of EDS systems for 13-odd years, I do have a few observations made as an analyst in the lab. I'm intentionally taking the devils advocate point of view here, and welcome corrections and comments.
First, those who used Tracor-Northern systems for EDS analysis obtained the Flextran code (interpreted Basic/Fortran sort of), and one could educate oneself as to exactly how you started with an EDS spectrum, obtained the fitted peak areas by least-squares, and finally got the concentrations of the elements after ZAF correction. When you bought the system, you got the source code. Today you will find that almost nobody will give you the very code that is used to do all this; you may find that it is not so easy to get explicit details of all the processing and correction algorithms (exactly whose version of ZAF or PRZ is being used?). In a nutshell, the software is not available to the user. The user can neither learn about all the details of the algorithm, nor add to it with code that they have written themselves (with the exception of macros or command sequences, which just automate procedures, mainly).
Secondly, most turnkey systems are demonstrated, purchased, and used to do so-called "standardless analysis", whereby either stored library EDS reference spectra are used (obtained typically on another instrument, likely at different take-off angle and accelerating voltage than what you are using to do your analyses), or fundamental parameters algorithms are used to calculate the spectra that would represent actual standards. Clearly, it is easier to acquire a spectrum on your sample and turn it over to the standardless software and let it do its magic. However, you implicitly accept errors possibly as large as 100% (!) due primarily to imperfections in the fundamental parameters equations and/or data. One is better off acquiring spectra on actual standards and fitting the sample spectra to those spectra acquired on the same instrument under the same conditions. But even in state of the art commercial systems, this procedure is convoluted and is definitely not within the realm of the casual user. And I have attended many a demo.
I hope we all agree that the goal is to perform quantitative EDS analysis. If not, then one can simply acquire spectra on different parts of (say) a failed solder joint, tell the client "looks like more Sn over here than over there" and be done with it. Many questions might be answered with this qualitative style of analysis. One can find commercial PC-based systems that allow you to acquire EDS spectra, but perhaps either don't do least-squares fitting and ZAF correction (cheaper systems), or only do the standardless thing (more expensive, generally). Many software packages do not even print out the fitted peak area and/or k-ratio (only the final analysis), and almost none propagate the counting statistics error from the peak measurement to the concentration units of the resulting analysis. For standardless analysis packages, many do not show you the normalization factor that was used, but we don't care 'cause those 100% totals every time sure look great. You know, users now pay up to several hundred dollars per hour for SEM/EDS time. Educated clients ask questions like "what is the concentration of Fe here and there, what is the precision of measurement for Fe vs. Mg, is Cr below the detection limit, and what is the accuracy of the analysis", and I'll bet that analysts outfitted with the current state-of-the-art systems can't answer those questions. (Not to say that older systems gave you this information either, it just has yet to be deemed as necessary information).
The trend is toward black-box configurations that are easier to use and produce a result that acceptable for typical applications -- understandably. EDS vendors have had to sell systems for a fraction of what they once sold for (and still carry on with development), and this places really tough constraints on what they can deliver. I admire them for toughing it out, and I know from dealing with EDS software that the code can get really complex and requires much work to develop. DTSA is a case in point; this program is arguably 5 years old and is still being tuned up because it is big big BIG.
My contention is that while hardware advances have been made, it is the software that is now hobbling quantitative EDS analysis.
Tell me this: who has an EDS system that can obtain a digital image of a sample where a full least-squares fit and ZAF correction has been done on the fly at each pixel, and the resulting image portrays the concentration of the element mapped (not a scaled x-ray intensity map). I've seen papers at national meetings describing this capability, as far back as 5-10 years ago. The limitation for sure isn't the processing speed of the computers now available, or the amount of RAM or hard drive storage space. How hard can it be to do a complete analysis at each pixel in an image?
Remember, our goal was to do quantitative EDS.
Paul Carpenter
+----------------------------------------------------+ | Paul K. Carpenter paulc-at-gps.caltech.edu | | Division Analytical Facility | | Geological and Planetary Sciences MC 170-25 | | California Institute of Technology | | Pasadena, CA 91125 | | 818-395-6126 (X-ray Lab) 818-568-0935 (Dept. FAX) | +----------------------------------------------------+
When Sorval got out of the microscopy business It sold off the 'Histology' line to Polaron/BioRad and the 'EM' line to RMC. I believe that if you contact Steven Slapp at 75767 -at- SMTP (ENERGY BEAM SCIENCES, INC) {75767.640-at-CompuServe.COM} he may be abel to help you. regards,
Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis University of North Carolina CB#7400, Rosenau Hall Chapel Hill, NC 27599-7400 Ph. 919-966-6343 Fax 919-966-6123
Thanks to those who have shown their interest in the
5 th. Brazilian Conference on Microscopy of Materials MICROMAT96 Rio de Janeiro October 13-16, 1996
Please note that there has been a change in the email address
sbme-at-dcmm.puc-rio.br
greetings to all Prof.Walter A.Mannheimer Metallurgy and Materials Engineering Federal University of Rio de Janeiro POBox 68505 21945 Rio de Janeiro, Brazil Voice 5521 280-7443 (Dept.office) 5521 590-0579 (direct) Fax 5521 290-6626 Email: wamann-at-metalmat.ufrj.br
Amen. We are discarding our (expensive, nearly useless and unsupported) commercial software and rewriting our old (Fortran) code that did most of what is described by Paul Carpenter.
On Tue, 21 May 1996, Paul K. Carpenter wrote:
} } It is nice to see a thread on energy-dispersive spectrometry get going } here. The original question dealt with selecting an EDS system from the } current systems available. There is no question that some really neat } advances have been made in the hardware arena for pulse processing etc. I } am not really qualified to comment on such things. } } However, as a user of EDS systems for 13-odd years, I do have a few } observations made as an analyst in the lab. I'm intentionally taking the } devils advocate point of view here, and welcome corrections and comments. } } First, those who used Tracor-Northern systems for EDS analysis obtained the } Flextran code (interpreted Basic/Fortran sort of), and one could educate } oneself as to exactly how you started with an EDS spectrum, obtained the } fitted peak areas by least-squares, and finally got the concentrations of } the elements after ZAF correction. When you bought the system, you got the } source code. Today you will find that almost nobody will give you the very } code that is used to do all this; you may find that it is not so easy to } get explicit details of all the processing and correction algorithms } (exactly whose version of ZAF or PRZ is being used?). In a nutshell, the } software is not available to the user. The user can neither learn about } all the details of the algorithm, nor add to it with code that they have } written themselves (with the exception of macros or command sequences, } which just automate procedures, mainly). } } Secondly, most turnkey systems are demonstrated, purchased, and used to do } so-called "standardless analysis", whereby either stored library EDS } reference spectra are used (obtained typically on another instrument, } likely at different take-off angle and accelerating voltage than what you } are using to do your analyses), or fundamental parameters algorithms are } used to calculate the spectra that would represent actual standards. } Clearly, it is easier to acquire a spectrum on your sample and turn it over } to the standardless software and let it do its magic. However, you } implicitly accept errors possibly as large as 100% (!) due primarily to } imperfections in the fundamental parameters equations and/or data. One is } better off acquiring spectra on actual standards and fitting the sample } spectra to those spectra acquired on the same instrument under the same } conditions. But even in state of the art commercial systems, this } procedure is convoluted and is definitely not within the realm of the } casual user. And I have attended many a demo. } } I hope we all agree that the goal is to perform quantitative EDS analysis. } If not, then one can simply acquire spectra on different parts of (say) a } failed solder joint, tell the client "looks like more Sn over here than } over there" and be done with it. Many questions might be answered with } this qualitative style of analysis. One can find commercial PC-based } systems that allow you to acquire EDS spectra, but perhaps either don't do } least-squares fitting and ZAF correction (cheaper systems), or only do the } standardless thing (more expensive, generally). Many software packages do } not even print out the fitted peak area and/or k-ratio (only the final } analysis), and almost none propagate the counting statistics error from the } peak measurement to the concentration units of the resulting analysis. For } standardless analysis packages, many do not show you the normalization } factor that was used, but we don't care 'cause those 100% totals every time } sure look great. You know, users now pay up to several hundred dollars per } hour for SEM/EDS time. Educated clients ask questions like "what is the } concentration of Fe here and there, what is the precision of measurement } for Fe vs. Mg, is Cr below the detection limit, and what is the accuracy of } the analysis", and I'll bet that analysts outfitted with the current } state-of-the-art systems can't answer those questions. (Not to say that } older systems gave you this information either, it just has yet to be } deemed as necessary information). } } The trend is toward black-box configurations that are easier to use and } produce a result that acceptable for typical applications -- } understandably. EDS vendors have had to sell systems for a fraction of } what they once sold for (and still carry on with development), and this } places really tough constraints on what they can deliver. I admire them } for toughing it out, and I know from dealing with EDS software that the } code can get really complex and requires much work to develop. DTSA is a } case in point; this program is arguably 5 years old and is still being } tuned up because it is big big BIG. } } My contention is that while hardware advances have been made, it is the } software that is now hobbling quantitative EDS analysis. } } Tell me this: who has an EDS system that can obtain a digital image of a } sample where a full least-squares fit and ZAF correction has been done on } the fly at each pixel, and the resulting image portrays the concentration } of the element mapped (not a scaled x-ray intensity map). I've seen papers } at national meetings describing this capability, as far back as 5-10 years } ago. The limitation for sure isn't the processing speed of the computers } now available, or the amount of RAM or hard drive storage space. How hard } can it be to do a complete analysis at each pixel in an image? } } Remember, our goal was to do quantitative EDS. } } Paul Carpenter } } } } +----------------------------------------------------+ } | Paul K. Carpenter paulc-at-gps.caltech.edu | } | Division Analytical Facility | } | Geological and Planetary Sciences MC 170-25 | } | California Institute of Technology | } | Pasadena, CA 91125 | } | 818-395-6126 (X-ray Lab) 818-568-0935 (Dept. FAX) | } +----------------------------------------------------+ } } }
As I said I used green phosphor from JEOL, cat. # 423-011. Call and see if this number still holds true.
You will need a collodion solution: 4 grams parlodian 25 ml absolute ethel alcohol 75 ml ether
You will also need a dish big enough for the screen and a cover (preferably glass) with a small hole for removal of the liquid.
1. Remove the old coating from the screen by washing in acetone. The cleaned plate must be free from all particles of matter and the surface must be free from blemishes and scratches.
2. Prepare a sufficient volume of 4% (w/v) suspension of phosphor powder in acetone containing about 1% collodion. The total volume must be sufficent to fill selected dish with liquid to a depth of about 1cm above the surface of the screen in position on the bottom of the dish.
3. Agitate the suspension vigorously (I used sonicator) then pour it rapidly into the dish.
4. Wait about 5-10 seconds to allow larger particles to settle and swirling to cease.
5. Slide the plate smoothly into the liquid, preferably without scraping the bottom of the dish.
6. Cover the dish and leave to settle. When the suspension has settled and the remaining liquid is clear, draw off the liquid by inserting a suction tube through the previously prepared hole in the lid of the dish. It is extremely important not ot disturb the screen plate or the liquid above it in any way as this is done. Draw off the liquid steadily and slowly then remove the suction tube.
7. Leave the screen to dry without any disturbance of any kind. Do not lift the lid to inspect the screen until the powder is quite dry because a slight change in drying conditions can produce a visible mark on the damp surface.
8. When the powder is quite dry, remove the screen and wipe off any excess phosphor from the back and sides of the screen.
9. Install.
10. A newly coated screen will outgas for a short time when it is first placed in the microscope. Pumping times may therefore be a little longer than normal at first.
A very small particle size is desirable for high resolution screens and it may be advantageous to agitate the the phosphor suspension in an ultrasonicator before putting it into the dish.
Hope this helps. It can get tricky and you may not get it the first time. Patience really helps!
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I have used a package produced by Oxford for the EXL series called QMAP that does perform a multiple least squares fit and quantitative analysis (bulk or thin film) on EDS spectra obtained at every pixel of a map. I have not used it for a few years, but it was very nice when trying to obtain maps from overlapped peaks in the EDS spectra.
I have no financial interest in Oxford, besides the fact that I am a customer.
Joe Michael _______________________________________________________________________________
It is nice to see a thread on energy-dispersive spectrometry get going here. The original question dealt with selecting an EDS system from the current systems available. There is no question that some really neat advances have been made in the hardware arena for pulse processing etc. I am not really qualified to comment on such things.
However, as a user of EDS systems for 13-odd years, I do have a few observations made as an analyst in the lab. I'm intentionally taking the devils advocate point of view here, and welcome corrections and comments.
First, those who used Tracor-Northern systems for EDS analysis obtained the Flextran code (interpreted Basic/Fortran sort of), and one could educate oneself as to exactly how you started with an EDS spectrum, obtained the fitted peak areas by least-squares, and finally got the concentrations of the elements after ZAF correction. When you bought the system, you got the source code. Today you will find that almost nobody will give you the very code that is used to do all this; you may find that it is not so easy to get explicit details of all the processing and correction algorithms (exactly whose version of ZAF or PRZ is being used?). In a nutshell, the software is not available to the user. The user can neither learn about all the details of the algorithm, nor add to it with code that they have written themselves (with the exception of macros or command sequences, which just automate procedures, mainly).
Secondly, most turnkey systems are demonstrated, purchased, and used to do so-called "standardless analysis", whereby either stored library EDS reference spectra are used (obtained typically on another instrument, likely at different take-off angle and accelerating voltage than what you are using to do your analyses), or fundamental parameters algorithms are used to calculate the spectra that would represent actual standards. Clearly, it is easier to acquire a spectrum on your sample and turn it over to the standardless software and let it do its magic. However, you implicitly accept errors possibly as large as 100% (!) due primarily to imperfections in the fundamental parameters equations and/or data. One is better off acquiring spectra on actual standards and fitting the sample spectra to those spectra acquired on the same instrument under the same conditions. But even in state of the art commercial systems, this procedure is convoluted and is definitely not within the realm of the casual user. And I have attended many a demo.
I hope we all agree that the goal is to perform quantitative EDS analysis. If not, then one can simply acquire spectra on different parts of (say) a failed solder joint, tell the client "looks like more Sn over here than over there" and be done with it. Many questions might be answered with this qualitative style of analysis. One can find commercial PC-based systems that allow you to acquire EDS spectra, but perhaps either don't do least-squares fitting and ZAF correction (cheaper systems), or only do the standardless thing (more expensive, generally). Many software packages do not even print out the fitted peak area and/or k-ratio (only the final analysis), and almost none propagate the counting statistics error from the peak measurement to the concentration units of the resulting analysis. For standardless analysis packages, many do not show you the normalization factor that was used, but we don't care 'cause those 100% totals every time sure look great. You know, users now pay up to several hundred dollars per hour for SEM/EDS time. Educated clients ask questions like "what is the concentration of Fe here and there, what is the precision of measurement for Fe vs. Mg, is Cr below the detection limit, and what is the accuracy of the analysis", and I'll bet that analysts outfitted with the current state-of-the-art systems can't answer those questions. (Not to say that older systems gave you this information either, it just has yet to be deemed as necessary information).
The trend is toward black-box configurations that are easier to use and produce a result that acceptable for typical applications -- understandably. EDS vendors have had to sell systems for a fraction of what they once sold for (and still carry on with development), and this places really tough constraints on what they can deliver. I admire them for toughing it out, and I know from dealing with EDS software that the code can get really complex and requires much work to develop. DTSA is a case in point; this program is arguably 5 years old and is still being tuned up because it is big big BIG.
My contention is that while hardware advances have been made, it is the software that is now hobbling quantitative EDS analysis.
Tell me this: who has an EDS system that can obtain a digital image of a sample where a full least-squares fit and ZAF correction has been done on the fly at each pixel, and the resulting image portrays the concentration of the element mapped (not a scaled x-ray intensity map). I've seen papers at national meetings describing this capability, as far back as 5-10 years ago. The limitation for sure isn't the processing speed of the computers now available, or the amount of RAM or hard drive storage space. How hard can it be to do a complete analysis at each pixel in an image?
Remember, our goal was to do quantitative EDS.
Paul Carpenter
+----------------------------------------------------+ | Paul K. Carpenter paulc-at-gps.caltech.edu | | Division Analytical Facility | | Geological and Planetary Sciences MC 170-25 | | California Institute of Technology | | Pasadena, CA 91125 | | 818-395-6126 (X-ray Lab) 818-568-0935 (Dept. FAX) | +----------------------------------------------------+
The old Dupont-Sorvall TC-2 Tissue Chopper unfortunately got lost in the shuffle when BioRad purchased their JB-4 and RMC their ultramicrotomes. It is no longer made, although used TC-2s can sometimes be found.
The McIlwain Tissue Chopper is distributed in the U.S. by Brinkmann Instruments (phone: 516-334-7500, fax: 516-334-7506).
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
I am trying to model a complex, interconnected membrane structure in three dimensions. If I can hand-measure dimensions off of micrographs, is there a computer program into which I can enter those numbers and then draw a "rotatable", 3-D image? I know that such a capability exists on some confocal systems but they have several million data bits to deal with. I would probably only have a hundred or so measurements. Am I asking the right question? Let me ask it this way: is there software available that would allow someone to scan TEMs of serial sections and then display a registered, 3-D image? Is this similar to what confocal scopes do?
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
After being used about 10 years, my Hitachi direct-drive rotary vacuum pump (model 160VP CuteVac for ISI DS-130 SEM) started making a rumbling noise this morning. The oil level is OK and the oil is clean. The SEM is functional but I think this is the first symptom of a future pump problem.
Does anyone know of any HItachi rotary pump suppliers in the US ? Thanks for your information.
we had problems with bubbles in the area of the specimen insertion of the workstation (our device is several years old, not one of the newer units). After playing around with it for several months, we bit the bullet and sent the sample holder back for assessment. Gatan told us it was a problem in the cryounit itself, not the workstation. From their description of the problem, our bubble problem is not an uncommon occurrence in the older units and was caused by a leak in the specimen arm. Our sampleholder in the same workstation worked fine after they rebuilt it and added new zeolite. The repair ran 1-2K.
good luck with it.
steve
} Hello everybody, } } we have problems with boiling nitrogen in the workstation of a Gatan } cryo holder. Particularly large bubbles form near the specimen making } transfer very difficult. Has anybody had that problem and found a } solution. } } Thanks in advance, } } Philip
Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University San Diego, CA 92182-4614 phone: (619)594-4523 fax:(619)594-5676 email:sbarlow-at-sunstroke.sdsu.edu
We have urgent need of an ion mill (used). We also find ourselves with a sum of money for equipment upgrade that we must either use or lose by June 30th. Please can anyone help?
} we have problems with boiling nitrogen in the workstation of a Gatan } cryo holder....
Before transferring the grid into the holder, we drain excess LN2 from the crytotransfer system by backing the stage a couple of centimeters out of the transfer device, until the flow of LN2 dribbling out of the port is low, then reseating the stage. The LN2 level is then just below the level of the stage, and the upper reservoir is still filled. We have encountered no problems with devitrification or frost when transferring the specimens to the stage through the cold nitrogen gas above the stage, so long as the transfer is made quickly with forceps that are adequately precooled (and kept cool by periodic immersion in the reservoir, should the grid need to be centered in the stage). We do this because we found that (a) loading was very difficult if the LN2 was at or above the level of the stage, and (b) the specimens often seemed to have more frost on them when they were transferred through the LN2.
} At 9:55 AM 5/21/96, melvyn dickson wrote: } } Distilled (and I imagine de-ionised) water has a more corrosive effect on } } copper piping than water with additive. Can't give a reference at this date } } but this was in the (engineering?) literature back in 1967. So we always use } } additives for fear of corroding cooling pipes inside lenses. } } } } That appeared to be the case for our JEM-4000EX. We use double distilled } water for the lens cooling. The cooper rust built-up was just unbelievable } we found out last year when we had to change the cooling water. The fluid } was so bad that I saved a vial for teaching purposes. I guess the ion } concentration in the fluid must be balanced so that it does not leave any } deposite and yet it does not try to take too many ions from the cooper } pipes. }
Perhaps another caveat regarding out use of de-ionized water. The system is very stable and we have not changed the water for years. Perhaps it has reached an equilibrium if corrossion of copper with de-ionized (not distilled) water is a problem. Also, there is some clear tubing in our system with no evidence of water discolouration.
alan.wilson-at-dsto.defence.gov.au
Dr Alan Wilson Senior Research Scientist Ship Structures and Materials Division Aeronautical and Maritime Research Laboratory Defence Science and Technology Organization 506 Lorimer St Fishermens Bend 3207 Victoria Australia ph 61 3 9626 7508, fax 61 3 9626 7816 or 61 3 9626 7087
} The classified ads in The Scientist include a lot of used equipment vendors. } The most recent issue also includes a web address (http://www.labx.com) and } indicates that you can find information from multiple vendors. I haven't } checked it out yet myself. } } Susan Udin } Dept. of Physiology } SUNY, Buffalo NY
Dear Long, I would imagine that the NSA (Nissei Sangyo America) company that sells Hitachi microscopes would sell the rotary pumps, since that pump is supplied on all their microscopes. I have had the same problem on my Hitachi rotary pumps and it turned out to be deterioration of the rubber coupler between the motor and pump. This was simple and cheap to fix myself and I keep a supply of them for when a pump gets noisy. If the pump starts to leak oil, then I replace the two shaft seals inside the pump, about a 1/2 hour job. The three pumps on my TEM have been going constantly for 131/2 years now. Regards, Mary Mager
Long Liang wrote: } Dear Microscopists, } } After being used about 10 years, my Hitachi direct-drive rotary vacuum } pump (model 160VP CuteVac for ISI DS-130 SEM) started making a rumbling } noise this morning. The oil level is OK and the oil is clean. The SEM } is functional but I think this is the first symptom of a future pump } problem. } } Does anyone know of any HItachi rotary pump suppliers in the US ? Thanks } for your information. } } Long Liang } ARCO EPMA/SEM Lab } Plano, TX
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
I should have been a little more thoughtful in extrapolating my experience with DSP's into the EDS arena. As Rick and Peter point out with only the experience EDS design engineers could, there are certain things mother nature just won't let us do.
However; I'll remain firm in my prediction that the digital front end will do more for EDS systems than the combined innovations of the last 10 years.
I'd like to introduce a new subject if I may. Would anyone care to comment on the ramifications to the end user if SEM manufacturers would agree to implement a standard interface for control of the beam by EDS systems?
Paul Carpenter and all listening to the EDS thread,
You've asked many good questions. It sounds like you've gotten quite a few frustrations off your chest at once! :)
I'm going to comment on just a few this time.
You write: I hope we all agree that the goal is to perform quantitative EDS analysis.
I'd respond, for some (mabey a majority) of us. I know of many situations in which it is sufficient to say what is present, even if I couldn't get the exact concentration. Also, in mapping situations, it's been sufficient to tell where something is, although I couldn't say exactly how much. I'd guess you must be involved in a critical process control application.
You raise some much larger questions regarding the evolution of systems and the accessability of an interested end-user to their inner workings. I think this could be a touchy issue with manufacturers, so I'm going to give it some thought before responding. But I agree with you that in general I don't like "point and shoot" type of instruments. This approach is nice in some situations, but us hard core users have lost some flexability. I'll broaden it a bit and ask: how could the SEM, EDS, and IA systems manufacturers provide some standardization (without suffering commercially) and provide end users with a common interface to control low level functionality of the various instruments?
Your last couple of paragraphs ring similar to a comment I made a few days ago. I think the manufacturers have been led around by users who are more interested in what operating system is being used than what the system can actually do. Not that having compatability with a particular computing environment isn't important, but it seems the evolution of some very nice software packages was cut short whilst everyone jumped on the Windows bandwagon and scrambled (spent a lot of programmers hours) to rewrite the code for more basic functionality of the system. Whew.
That's probably why PDP-11 based systems hung around for so long. I'll bet you can still find PDP-11 based EDS systems that can do a lot of things many of the new Windows and Mac based systems can't. How do we avoid this waste the next time around? Mabey all EDS and SEM related software can be written in JAVA.
} To all, } } I am trying to model a complex, interconnected membrane structure } in three dimensions. If I can hand-measure dimensions off of micrographs, } is there a computer program into which I can enter those numbers and then } draw a "rotatable", 3-D image? I know that such a capability exists on } some confocal systems but they have several million data bits to deal with. } I would probably only have a hundred or so measurements. Am I asking the } right question? Let me ask it this way: is there software available that } would allow someone to scan TEMs of serial sections and then display a } registered, 3-D image? Is this similar to what confocal scopes do?
Robert -
I use NIH Image to accomplish some of these tasks (serial TEM section reconstruction, distortion correction, etc), starting with drawing outlines from scanned TEM negatives. Image comes with built in registration capabilities, but you easily can enhance the functionality (and interactivity needed) with macros. Lets say, you generate a macro, which allows you to registrate/measure some points on your image (thresholded, manually drawn, whatever), save these measured points as coloured points on extra images, have these images a few times duplicated (always in mind the scale calibration) to get real 3d objects of your measured points, have them projected with lots of angle and transparency paramters, and save them as pics image stacks. Animate them. Very handy. However - Image does NOT allow no realtime interactive rotation of the projected images.
Dietmar
+++ Dietmar Reiter {dietmar.reiter-at-uibk.ac.at} ++++++++++++++++ +++ Dept. of Zoology and Limnology, University of Innsbruck ++++ +++ Technikerstrasse 25, A - 6020 Innsbruck, Austria +++++++++ +++ ph: (+43)-512-507-6170 (-6161), fax: ..-507-2930 (-2957) +++
Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis University of North Carolina CB#7400, Rosenau Hall Chapel Hill, NC 27599-7400 Ph. 919-966-6343 Fax 919-966-6123
I recently was assigned to investigate the utility of confocal and microprobe (perhaps AFM) microscopy for examining topical applications of biocides on pig skin samples. I have already performed some SEM/EDS work on this but now the question is how does the biocide affect the skin surface. I was hoping someone may know of some similar work in this area.
Thanks in advance,
Dave Audette Olin Research Center Cheshire, CT deaudette-at-corp.olin.com
Subject: Time: 10:09 AM OFFICE MEMO RE:Corrosion w Deionized water Date: 5/23/96
I would be very surprised if either deionized or distilled water is significantly more corrosive than ordinary tap water in copper and stainless steel systems. In a wet environment corrosion (i.e. the dissolving of the metal) always is a galvanic phenomenon that involves a potential difference between two parts of the system, an anodic area and a cathodic area. The anodic area is always more electrochemically reactive than the cathodic area, and the overall corrosion mechanism consists of four separate processes 1. Metal atoms dissolve at the anodic area, forming metal ions which go into solution. 2. This reaction releases electrons, which travel through the metal parts to the cathodic area. 3. At the cathodic area these electrons are consumed in one or more of a variety of 'cathodic reactions' (in aerated, neutral solutions such as might typically be found in water chillers this reaction usually involves the combination of 2 electrons with two molecules of water and one molecule of oxygen to form 4 hydroxyl ions). 4. There also must be a flow of ions between the two electrode areas through the water that connects them to maintain charge neutrality. Corrosion can be retarded by interrupting any one of these four processes, and can be enhanced by anything that promotes any of them. One common cause of accelerated corrosion is having two metals of considerably different electrochemical activity directly in contact in an aqueous environment. An example is putting a steel pipe fitting into a copper water line. The iron is electrochemically more active than the copper, and so it will become strongly anodic, releases iron ions into solution, and corroding away rapidly. (The iron ions react with hydroxyl ions in solution forming a hydrated oron oxide, which we call rust. Rust is evidence of corrosion, but not the basic problem.) If you put plastic nipples between the iron and copper components so that there cannot be electron flow from the iron to copper, corrosion will be prevented. I suspect that systems filled with deionized or distilled water that show high rates of discoloration, and other evidence of corrosion, have such bimetallic couplings that are promoting the corrosion. Under some circumstances stainless steel is cathodic with respect to copper, and so if a system has stainless steel parts directly connected to copper parts, so that there could be a flow of electrons between them, then sufficient galvanic potential might develop between them to cause the copper to corrode (i.e. copper ions might dissolve into the solution and cause it to become discolored). I think this is a more likely explanation for any such phenomena than the lack of ions in solution in the deionized and distilled waters, and I would expect that the phenomena would disappear if the system were rearranged so that the copper and stainless steel parts were no longer in direct contact. Ref: Materials Science & Engineering by W. D. Callister, 2nd Ed., Wiley, Ch. 18). W. C. Bigelow (bigelow-at-umich.edu)
Thanks to all for the information on computer programs for 3D reconstructions. Looks like NIH Image is the easiest and cheapest way to get started in this.
And I was mightily impressed by the 600-700 consecutive serial sections reported by one respondent!
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
} } Colleagues: } } I am looking for a source for DNP conjugated secondary antibodies. } Can anyone help?? } ******************************************************* Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
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Hi there,
On several occasions I have been asked for literature on the how's and why's of EDX analysis. Does anyone have any suggestions as to where I can obtain such material?
Microscopists, we have had less than favorable results fixing buffy coat of reptiles (white blood cell fraction) with standard 2.5% glute in 0.1M cacodylate. Unfortunately we do not know the osmolarity of the blood, though our experience with other reptiles reveals a significant range within the same species (wild caught). Does anyone have any suggestions such as possible additives?
Thanks in advance,
ERJ Elliott Jacobson Professor Department of Small Animal Clinical Sciences P.O. Box 100126 College of Veterinary Medicine University of Florida Gainesville, Florida 32610, USA Phone: 904-392-4700 X4773 Fax: 904-392-6125 E-Mail: ERJ-at-vetmed1.vetmed.ufl.edu WEB Site: www.vetmed.ufl.edu
In article tania-at-dynamotive.com (Tania Jones) writes: } Date: Thu, 23 May 1996 15:11:18 -0700 } From: tania-at-dynamotive.com (Tania Jones) } Subject: EDX information
} Hi there,
} On several occasions I have been asked for literature on the how's and } why's of EDX analysis. Does anyone have any suggestions as to where I } can obtain such material?
If its still available a very clear intoductory teatment is:
Energy Dispersive X-Ray Microanalysis: An Introduction Kevex Instruments Inc., San Carlos CA Douglas Vaughan, Ed. printed 1983, 1988, 1989, 199?
Also X-Ray Microanalysis in the Electron Microscope J.A. Chandler 1977 a volume in Practical Methods in Electron Microscopy Audrey Glauert, ED. Elsevier/North-Holland ISBN 07204 0607 2
A much fuller treatment: Scanning Electron Microscopy and X-Ray Microanalysis 2nd Edition Goldstein, Newbury, Echlin, Joy, Romig, Lyman, Fiori and Lifshin Plenum Press ISBN 0-306-44175-6
`On several occasions I have been asked for literature on the how's and `why's of EDX analysis. Does anyone have any suggestions as to where I `can obtain such material? ` `Thanks in advance, ` `Tania Jones `Lab Manager `DynaMotive Technologies
I have been writing a series of articles on EDX for the newcomer for Microscopy Today entitled "More than one ever wanted to know about x-ray detectors." These are available from Microscopy Today, from me, or maybe best of all, on http://www.MOXTEK.com.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. ************************************************* Orem UT 84057 **"Soft x-rays in the 21st Century" conference ** 801-225-0930 ** 8-11 January 1996, Midway Utah ** FAX 801-221-1121 ** http://volta.byu.edu/xray/info.html ** lundm-at-xray.byu.edu *************************************************
"He spoke with a certain what-is-it in his voice, and I could see that, if not actually disgruntled, he was far from being gruntled."
Comments: Authenticated sender is {wesleysm-at-biology.und.ac.za}
Hi Rajesh We have routinely prepared Aspergillus small plugs for TEM using an overnight fixation in 2.5%GA in phosphate buffer, followed by several buffer washes and postfixation in 0.5 - 1% osmium tetroxide for 1 hour (all at 4C). After a distilled-water rinse, en bloc staining/fixation with 0.5% uranyl acetate in 30% acetone (30 min), followed by graded acetone series, infiltrated with 50% Spur's resin for 4 hs and overnight in full resin.
Two questions to the forum, though:
1. It is interesting that the cultures will float until (approx) the 50% acetone step. Although osmium vapour will fix those exposed areas, I wonder about the role played by GA (or even U Ac) when it comes to conidiophores and conidia. The results are good anyway.
2. I have know users to favour the use of an equal-parts mixture of GA and osmium at fairly high concentrations for SEM. How widespread is the use of this protocol? Given the instantaneous cross-reactivity between the two fixatives, I really wonder about its usefulness.
Any comments?
James Wesley-Smith Electron Microscope Unit George Campbell Building University of Natal Durban, South Africa
Dear Paul, In regard to your question, I would like to point out that the Oxford Instruments Link ISIS system has a fully quantitative mapping package (Quantmap). Full spectrum processing (including digital filtering and fitting) followed by a quantitative matrix correction is performed at each point. The resulting digital images show elemental concentrations rather than just a scaled x-ray intensity map. Standard deviation maps are also produced to allow significance testing.
Your local Oxford rep would be delighted to tell you more! Best Regards, Julie Sheffield-Parker.
At 05:15 PM 21/05/96 -0700, you wrote:
} } Tell me this: who has an EDS system that can obtain a digital image of a } sample where a full least-squares fit and ZAF correction has been done on } the fly at each pixel, and the resulting image portrays the concentration } of the element mapped (not a scaled x-ray intensity map). I've seen papers } at national meetings describing this capability, as far back as 5-10 years } ago. The limitation for sure isn't the processing speed of the computers } now available, or the amount of RAM or hard drive storage space. How hard } can it be to do a complete analysis at each pixel in an image? } } Remember, our goal was to do quantitative EDS. } } Paul Carpenter } } } } +----------------------------------------------------+ } | Paul K. Carpenter paulc-at-gps.caltech.edu | } | Division Analytical Facility | } | Geological and Planetary Sciences MC 170-25 | } | California Institute of Technology | } | Pasadena, CA 91125 | } | 818-395-6126 (X-ray Lab) 818-568-0935 (Dept. FAX) | } +----------------------------------------------------+ } } } }
Are there any microprobe (WDS) people who analyze Os alloys (e.g., osmiridium)? If so, what standard are you using, and where did you obtain it?
----------------------------------------- Alfred Kracher Geological Sciences Iowa State University Ames, IA 50011-3212 akracher-at-iastate.edu http://www.public.iastate.edu/~akracher vox:515 294 5439 fax:515 294 6049 -----------------------------------------
} } Are there any microprobe (WDS) people who analyze Os alloys (e.g., } osmiridium)? If so, what standard are you using, and where did you obtain } it? } Dear Alfred, I am not doing WDS, nor am I studying alloys, but I made some Ir and Pt standards by using the acetylacetonate complexes, which dis- olve in resin. These are available from Strem Chemicals Inc., 7 Muliken Way, Dexter Industrial Park, Newburyport MA 01950. Alfa Aesar, (800) 343-0660, carries Os, Ir & Pt in many forms--powder, wire, sponge, etc. I guess these would be better suited for stds for alloys. Yours, Bill Tivol
Aspergillus freezes quite nicely. If you have the set-up for plunge freezing and freeze substitution I would recommend that procedure over regular chemical fixation. I can send you the protocol if you want to consider fs.
Best regards,
Beth Richardson Botany Dept. EM Lab Coordinator
} Does anyone have a working protocol for processing aspergillus? } } thanks in advance. } } rpatel-at-rwja.umdnj.edu
Does anyone know where I can purchase TEM apertures (condenser, objective, and selected area) for a Philips EM301 (besides Philips).
Thanks
************************************************************************** Lucille A. Giannuzzi, Ph.D. phone: 407 823-5770 University of Central Florida fax: 407 823-0208 Materials Science Program email: lag-at-pegasus.cc.ucf.edu Dept. of Mechanical and Aerospace Eng. Orlando, FL 32816-2450 **************************************************************************
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A list of companies that broker or directly sell used microscopes and accessories is available on the WWW at http://www.mwrn.com/product/used.htm
Susanne Pignolet Brandom, Ph.D. MC Services
MicroWorld Internet Resources at http://www.mwrn.com/ MicroWorld News available by e-mail from MWN-at-mwrn.com
} At 01:39 PM 5/16/96 -0500, you wrote: } } I am looking for sources of used equipment such as equipment brokers etc. } } Specifically the types of equipment include coating, microscopy and } } mechanical test equipment. Secondly I'm also trying to locate sources of } } discounted consumable supplies for metallurgical polishing work. } } } } I would deeply appreciate any leads. } } } } } } }
Message-ID: {199605241808.NAA01386-at-IndyNet.indy.net} To: "Audette, David" {deaudette-at-corp.olin.com} , Microscopy list {microscopy-at-Sparc5.Microscopy.Com}
On Thu, 23 May 1996, Tania Jones wrote:
} Hi there, } } On several occasions I have been asked for literature on the how's and } why's of EDX analysis. Does anyone have any suggestions as to where I } can obtain such material? } } Thanks in advance, } } Tania Jones } Lab Manager } DynaMotive Technologies
In the recent thread on this topic, Eugene Betrin's book, "Principles and Practice of X-ray Spectrometric Analysis" was mentioned (ISBN 0-306-30809-6). I checked out a copy from our Chemistry department library. It is extremely comprehensive, almost to a fault. Another good reference source is Joseph Goldstein, etal "Scanning Electron Microscopy and X-ray Microanalysis" (ISBN 0-306-44175-6). Both of these are texts, so might be a little deep for the casual reader.
A week or so ago I presented a question how to prepare insect eggs for TEM. I thank everyone who sent us advice in this area, however we are still unable to successfully infiltrate the eggs. So far this is what has been done.
1) Standard fixation and infiltration protocol. 2) Microwave fixation for impervious biological specimens. 3) Pre-treatment with 1M meta periodate, extended acetone dehydration followed by infiltration with Spurrs. All of these methods left us with collapsed samples.
I would like to try a pre-treatment of chitinase and possibly a combination of chitinase and meta periodate on this sample. Does anyone out there in the microscopy world have experience with the area or know of a reference?
Once more Thank you
---------------------------------------------------------------------------- --------- Karen Vaughn Tel.(904) 392-1184 EM Technician University of Florida Fax.(904) 846-0251 Electron Microscopy Core Laboratory Email. KLV-at-biotech.ufl.edu Interdisciplinary Center for Biotechnology Research http://www.biotech.ufl.edu/~emcl/ 214 Bartram Hall Gainesville, Fl 32611
As promised, following is the summary of responses I received to my = request for information on sputter coaters.
There were a total of 21 responses. 17 of those contained evaluations: = 8 Denton Desk II, 3 SPI Sputter Module, 1 Ernest F. Fullam EFFACoater, 1 = each for Edwards EL2E, Hummer V, Ladd evaporator, and 2 who asked not to = be included in the summary and have not been.
There were a couple themes that ran through virtually all the responses. = Almost everyone 1) was happy with what they had, and 2) felt what they = had was better than what it replaced.
Out of the 18 responses there were two comments about the current = control response via vacuum control being slow (one each Denton and = SPI). And one about a slightly mis-shaped glass chamber cylinder which = required paying a little attention to orientation to insure a good = vacuum seal (Denton). All other comments were of a positive nature. = Judging from the total of the comments received, it would appear the = units available are all reasonably reliable. There were several = comments about service, all good.
There were 8 responses on film thickness monitors. The "vote" was 5 = "not necessary" to 3 "necessary". The general feeling seemed to be = that FTM's were not all that worthwhile primarily because there were too = many factors affecting their correlation to the metal actually deposited = on the sample.
I can provide the individual responses (less the two who requiested to = be excluded) to anyone interested.
My thanks and appreciation to all those who took the time to respond.
Marc C. Brande, MS, Founder Cultured Cell Systems Voice: (619) 587-4830 3840 Camino Lindo FAX: (619) 552-1516 San Diego, CA 92122 Email: mcbrande-at-sierra.net
} Dear Paul, } In regard to your question, I would like to point out that the Oxford } Instruments Link ISIS system has a fully quantitative mapping package } (Quantmap). Full spectrum processing (including digital filtering and } fitting) followed by a quantitative matrix correction is performed at each } point. The resulting digital images show elemental concentrations rather } than just a scaled x-ray intensity map. Standard deviation maps are also } produced to allow significance testing. } } Your local Oxford rep would be delighted to tell you more! } Best Regards, Julie Sheffield-Parker. } } At 05:15 PM 21/05/96 -0700, you wrote: } } } } } Tell me this: who has an EDS system that can obtain a digital image of a } } sample where a full least-squares fit and ZAF correction has been done on } } the fly at each pixel, and the resulting image portrays the concentration } } of the element mapped (not a scaled x-ray intensity map). I've seen papers } } at national meetings describing this capability, as far back as 5-10 years } } ago. The limitation for sure isn't the processing speed of the computers } } now available, or the amount of RAM or hard drive storage space. How hard } } can it be to do a complete analysis at each pixel in an image? } } } } Remember, our goal was to do quantitative EDS. } } } } Paul Carpenter } ************************************************* } From:- } } Julie Sheffield-Parker, } Oxford Instruments Pty. Ltd., } P. O. Box 7, } Pennant Hills, } NSW 2120, } Sydney, AUSTRALIA } } Tel: ++ 61 2 484 6108 } Fax: ++ 61 2 484 1667 } E-Mail: oisydney-at-ozemail.com.au } } *************************************************
Hi,
With respect, I don't think the issue is, for example, the capability to do quantitative EDX mapping. Surely the question is how? Commercial EDX systems are just (software) black boxes. Before anybody can have real confidence, it is at least necessary to know the algorithms used for the data processing, although the source code would be preferable.
However, having been involved in EM for some time, I would agree with an earlier comment, that for a large number of microanalytical problems, true quantitative analysis is not required - knowing that there is ' a lot', 'some', or 'a trace' of a certain element at a specific point is usually all the information that is required.
=46IVE POSTDOCTORAL POSITIONS IN MATERIALS PHYSICS
OAK RIDGE NATIONAL LABORATORY, SOLID STATE DIVISION
=46ive postdoctoral openings are available in the Electron Microscopy Group, ORNL, in a joint experimental and theoretical program coupling atomic scale imaging of interfaces with state of the art computational studies on interface structure and properties. The Solid State Division has one of the world's finest facilities for atomic scale imaging of materials: a VG Microscopes HB603 300 kV scanning transmission electron microscope with a 1.26=C5 probe size provides direct, Z-contrast imaging capabilities for interfaces in materials. A VG Microscopes HB501UX 100 kV microscope with a high sensitivity parallel EELS capability provides atomic resolution spectroscopy. The Electron Microscopy Group has a Silicon Graphics PowerIndigo workstation with a Molecular Simulations' Cerius 2 package incorporating the CASTEP pseudopotential code. The Division has a number of additional workstations, and access to extensive parallel computing capabilities, the Intel Paragon XP/S 35 and XP/S 150 with 512 and 2048 processors respectively.
In addition, postdoctoral positions are available in collaboration with the Ion-Solid Interactions Group, ORNL, Northwestern University and the University of Illinois at Chicago:
(I) Grain Boundary Structure and Properties:
Two positions are available in the Solid State Division, ORNL (Dr. Steve Pennycook and Dr. Richard Wood). Atomic scale imaging and spectroscopy, macroscopic property measurements and theoretical simulations will unravel the link between grain boundary atomic structure and the electrical transport properties of electronic ceramics and superconductors. The program will focus on bicrystals and thin films, and investigate the applicability of the structural unit model for predicting properties, its extension to three dimensions, and impurity site imaging and spectroscopy with a view to grain boundary engineering. Theoretical studies will range from empirical bond-valence sums to full ab initio calculations. Two positions are available, either one theorist and one experimentalist, or alternatively, qualified candidates may wish to perform theory and experiment on a specific class of materials.
(II) Interfaces in Functional Magnetic/Electroceramic Materials:
This position is a collaborative venture between the Solid State Division (Dr. Steve Pennycook) and The Department of Materials Science & Engineering, Northwestern University (Prof. Vinayak P. Dravid), focussing on the role of interfaces in functional magnetic and electroceramic materials, e.g. colossal magnetoresistant (CMR) materials, PTCR, GBBLC oxides. Atomic resolution Z-contrast imaging and EELS will be used to determine the atomic and electronic structure of interfaces in thin film multilayers and bulk polycrystals for correlation with macroscopic properties. The successful applicant will reside at ORNL for at least 2/3 of time, and serve as a liaison between Northwestern University and ORNL. Instrumentation available at NU in the newly restructured Electron Probe Instrumentation Center (EPIC) includes a Hitachi HF-2000 FEG TEM/STEM with x-ray, EELS, in-situ I/V cryogenic holder and e- holography capability, and a Hitachi S4500-II FEG SEM with x-ray, EBSP/OIM and liquid helium stage with in-situ I/V probes. This position also involves collaborative work on other interface controlled materials such as electroceramics and high Tc compounds.
(III) Structure-Property Relationships in MBE grown Optoelectronic Devices:
This position is a collaborative venture between the Solid State Division (Dr. Steve Pennycook) and the Department of Physics, University of Illinois Chicago (UIC, Prof. Nigel Browning), to investigate the fundamental link between the properties of MBE grown optoelectronic devices and the atomic structure of the film-substrate interface, leading ultimately to the production of compliant and alternative substrates for coherent heteroepitaxy. Z-contrast imaging and electron energy loss spectroscopy offer unique insights into the nature of the compliant interface, and film and defect nucleation mechanisms. The Microphysics Laboratory at UIC is internationally recognized for its extensive facilities and expertise in MBE growth, including an OPUS 45 MBE facility for the transfer and processing of 5-inch wafers, and two RIBER 2300 MBE systems with extensive growth monitoring and in-situ surface analysis capabilities. This position offers a unique opportunity to work at a fundamental level in an area of high technological significance. Applicants should demonstrate extensive knowledge of crystal defects.
(IV) Ion-Solid Interactions:
Research in the Ion-Solid Interations Group is materials oriented and involves ion beam synthesis or modification of materials to produce novel and technologically relevant properties. Research in which the successful candidate will be involved includes (a) the formation of nanocrystals and quantum dots in insulators to produce unique optoelectronic responses, (b) synthesis of buried layers in semiconductors for interconnect or isolation applications, and (c) the physics of ion-induced defects in semiconductors. The diverse scope of this program and the interactions it has fomented with other national laboratories, industry, and universities will provide the successful candidate a unique opportunity for professional growth. Applicants should have extensive experience in transmission electron microscopy and the preparation of cross section specimens.
Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound background in the relevent materials issues and a burning ambition to develop a forefront area in materials physics. If this is you, send your resume and publication list to Dr. S. J. Pennycook at the address below. Prior experience using transmission electron microscopy is essential only where explicitly stated; consideration will be based on the candidates overall potential for success in the field. Positions are for one year initially, normally renewed for a second year and possibly a third. ORNL is a multipurpose national laboratory managed by Lockheed Martin Energy Research Corporation for the U.S. Department of Energy. ORNL is an equal opportunity employer committed to building and maintaining a diverse work force.
---------------------------------------------------------------------------- ------------------------------------------Stephen J. Pennycook Oak Ridge National Laboratory PO Box 2008 Oak Ridge TN 37831-6030
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-- [ From: Charles A. Garber., Ph.d * EMC.Ver #2.10P ] --
Forgive me if this is thought to not be an appropriate use of the listserver, but I just found out today that Dr. Wilf Gee, VG Microtech UK (Fisons Instruments) is retiring the end of the month.
Dr. Gee for many years was the chief engineer (and he held other high positions as well) at what was once Polaron Equipment Ltd, and which later was acquired by Bio-Rad. That business was sold to Fisons in about 1992.
It is my understanding that Dr. Gee might very well be the only person who really knows the inner workings of some of the larger equipment items, especially that which was made some years ago by Polaron Equipment. Certainly more than a few persons including yours truly have grown to depend on Wilf's advice and guidance when it came to keeping in operation equipment made some years ago.
Wilf was willing to have me post this information as well as his e-mail address: {wgee-at-vacgen.fisons.co.uk} . Of course, the e-mail address is probably going to "expire" at the end of the month as well, so anyone wanting to say in touch with him should send him their address before that time. Wilf, always wanting to help someone in need with their equipment, I am sure, even in his retirement, would want to pass on what he knows.
At least I for one wish Wilf a happy and healthy coming retirement!
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: spi2spi-at-2spi.com
A research associate is sought to establish, operate, and maintain the Environmental Scanning Electron Microscope which is to be set up as a University user and outreach facility. The individual is expected to conduct research on materials characterization and behavior with the ESEM in collaboration with faculty, students and government personnel. Research areas include composite materials, metals, ceramics, polymers, fiber reinforced cement, asphalts, food, packaging materials, soils, plants and wood products. The ESEM is equipped with various stages for in situ stress-temperature-environment studies over a wide range of conditions. The ESEM is also equipped with an EDS detector for chemical analysis.
The person occupying this position will be responsible for operation and maintenance of the ESEM as well as the training of faculty, staff, graduate students and non-campus users. In addition, the successful candidate will conduct research with the ESEM and is expected to develop contacts with potential off-campus users in the local and state-wide community to assist in generating funds for the support and maintenance of the ESEM. The responsibilities for this position are divided approximately as follows: Research-50%, Outreach-25%, Administration-15%, Teaching-10%.
Qualifications: A Ph.D. in Materials Science, Physics, Chemistry, Geology, Biology or Engineering is required. A combination of course work and hands-on research experience with SEM is required, ESEM is preferred. The applicant must exhibit very high levels of oral and written communication skills. Previous experience in budgeting and accounting would be useful.
Position is open until filled. Salary is commensurate with qualifications and experience. Send a complete curriculum vitae, graduate level transcript, and three references to: Professor Lawrence T. Drzal, Michigan State University, Composite Materials and Structures Center, Engineering Research Complex B-100, East Lansing, MI 48824-1326 tel: 517/ 353-5466 fax: 517/ 432-1634
Contact Michael Rich, Research Specialist and Laboratory Manager, for further information regarding this position. RICH-at-EGR.MSU.EDU. Michigan State University is an equal-opportunity institution.
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-- [ From: Charles A. Garber., Ph.d * EMC.Ver #2.10P ] --
The concept of CIC, which I had not heard of previously, was described as follows: ==================================== "CIC Agency is a Risk Management Corporation .....essentially an insurance company." ====================================
This discussion is beginning to sound somewhat like our national debate on health care and the HMO approach in that people are a) complaining about the ever increasingly higher costs of providing microscope care, b) have a perception that the people offering such contracts for this care are taking advantage of what is thought to be a captive market, and c) are under no financial incentives (e.g. the patient microscope lab managers) to help reduce the costs for the provision of the services on the part of the service provider.
I can remember some years ago having this very samediscussion with the National Service Manager of one of the major column instrument manufacturers in the USA. He said "legally" it could never happen because of "GSA Contract considerations" as well as other reasons. I presume he was talking about it from the standpoint of their particular firm offering service contracts at different prices to different people, since the pricing would be "experience based". There is of course a concept that one can not sell something at a lower cost than they would sell the same thing for to Uncle Sam. On the other hand, maybe he just did not understand what I was suggesting. He might even have been incorrect in his legal interpretation and whether the GSA concern would apply in this kind of a situation.
He went on to say "the approach you are suggesting is already in place, our people when they visit a customer spend part of their time looking for the embryo of tomorrow's problem on today's visit because it in our own self-interest to keep service calls to a minimum". Well, that could certainly be correct, but as has been found to be the case for the provision of medical services, there are many aspects of one's health that just are not apparent during that brief visit and exam. And the patient has to take ultimate responsibility for watching for signs of anything not being right. Yet at the same time, because of human nature being what it is, the existence of a "co-pay" is a vital component of any kind of cost containment process.
And in today's environment, service engineers seem to be so harried, and so pressed to increase the number of service calls made per unit of time, that I am not sure just how much discretionary time is really left any more to seek out those "problems just waiting to happen".
So unless firms like CIC are going onto the premises of their client firms, holding training sessions and seminars on how one could operate their laboratory from the perspective of reducing the costs to the insurer, so to speak, then CIC is indeed acting only as an insurer, and is NOT acting as a provider of a broad risk management program for the laboratory (one that has as the goal to help that laboratory client to operate in a way that the costs to CIC would be less rather than more, the incentive being a lower premium in coming years). And only a believer in the tooth fairy would also believe that somehow, when it was all said and done, after taking into consideration this extra layer of bureaucracy, not to mention corporate profit, that they were going to be getting the exact same service for less money.
And this is because just like with an HMO, there are only a limited number of ways the "insurer" can reduce costs:
a) limit or ration benefits (for example, agree to perform only a fixed number of "emergency" calls in one year) under the Agreement. Maybe routine service calls would be scheduled more at the convenience of the service provider rather than the convenience of the customer. There could be other accommodations that might mean a lot to the insurer but not represent much of a "give back" to the laboratory.
b) institute a meaningful co-pay, so that each time a service engineer is called in to work his charm, there is a definite "out of pocket" cost paid, thereby limiting (just as it is meant to do for visits to a physician's office) the total number of visits. In our own laboratory situation, it is just amazing how much that extra incentive to solve problems on the phone instead of calling for an emergency visit can reduce the actual number of required visits.
c) institute a real program of loss prevention and risk analysis, where by on a regular basis, the laboratory is visited, and "hazardous" (with regard to future equipment failures) situations are spotted and remedied before causing the insurance company an expensive "loss". One does not have to eliminate too many emergency field service calls before such a program would provide large economic rewards. Although I have no hard data, but just the experience of managing column instruments for nearly thirty years, just spotting a water chiller about to konk out or a pump belt getting ready to break could save a great deal of down time and money.
We ourselves made a comparison between the frequency of needed calls for service engineers before vs. after I dropped all annual service contracts in our own laboratories. It was amazing how many fewer visits we seem to be able to get along with, without any noticeable increase in down time. One might think that as instruments get older, then they would require more service, but for us, that has not been the case. Perhaps it is the pedigree of our instruments. The several major disasters we have had, although expensive, still leave us light years ahead of where we would have been had we been covering the instruments under paid service contracts (at one time six) all these years. And the "disasters" and ensuing down time would have happened anyhow, service contract or no service contract. But those are the outliers of experience that do occasionally occur and unfortunately too often become the horror stories on which the sales of service contracts are promoted.
If some agency was going to establish some kind of insurance company that at the same time, would institute a good risk management approach to the servicing of column instruments, EDS systems, etc., then I would sure like to know about it myself. We would be among the first to want to sign up. We would see that as a really worth while program since it would have a real chance of reducing the costs to maintain a column instrument.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: spi2spi-at-2spi.com
Hello, I was away for a few days and noticed much discussion about cooling water. If I may, at this late date, I would like to spalsh around a bit myself.
Many years ago, I was a Service Engineer for Philips and had about 15 Haskris systems running in Florida. My philosophy has always been Simplicity in concept and exicution or the well known "KISS Principal." Anyway, the formula for sucess was to use dionized water or distilled water, put three or four drops of oil on top of the water, in the reservour and if you have some, toss a couple crystals of Iodine in the tank. A five micron water filter is a good idea, also. The way it works, as I understand it...Since water is such a wonderful solvent, it leaches away a small bit of the material through which it travels and creates a eguilibrium. The water sort of tunes itself to the system. A fungicide or other poison does what it should but what is the result? My experience has been the creation of gray slime in the system. A monolayer of oil on the reservour keeps algae away. Additives have been the cause of many problems with seals, small openings where the water is supposed to flow and pump failures.
Thanks for your time.
Alex Greene Scientific Instrumentation Services, Inc. Austin, Texas
Hi Phil, I always wondered how to coat a screen. Thanks very much for the detailed information. For anyone not up to trying it themselves, Grant Scientific does a great job of screen recoating. They are located in Gilbert, South Carolina and their phone no. is 803/892-2841.
Alex Greene Scientific Instrumentation Services, Inc. Austin, Texas
At 11:23 AM 5/22/96 -0400, you wrote: } To all interested: } } As I said I used green phosphor from JEOL, cat. # 423-011. Call and see } if this number still holds true. } } You will need a collodion solution: 4 grams parlodian } 25 ml absolute ethel alcohol } 75 ml ether } } You will also need a dish big enough for the screen and a cover } (preferably glass) with a small hole for removal of the liquid. } } } 1. Remove the old coating from the screen by washing in acetone. The } cleaned plate must be free from all particles of matter and the surface } must be free from blemishes and scratches. } } 2. Prepare a sufficient volume of 4% (w/v) suspension of phosphor powder } in acetone containing about 1% collodion. The total volume must be } sufficent to fill selected dish with liquid to a depth of about 1cm above } the surface of the screen in position on the bottom of the dish. } } 3. Agitate the suspension vigorously (I used sonicator) then pour it } rapidly into the dish. } } 4. Wait about 5-10 seconds to allow larger particles to settle and } swirling to cease. } } 5. Slide the plate smoothly into the liquid, preferably without scraping } the bottom of the dish. } } 6. Cover the dish and leave to settle. When the suspension has settled } and the remaining liquid is clear, draw off the liquid by inserting a } suction tube through the previously prepared hole in the lid of the } dish. It is extremely important not ot disturb the screen plate or the } liquid above it in any way as this is done. Draw off the liquid steadily } and slowly then remove the suction tube. } } 7. Leave the screen to dry without any disturbance of any kind. Do not } lift the lid to inspect the screen until the powder is quite dry because } a slight change in drying conditions can produce a visible mark on the } damp surface. } } 8. When the powder is quite dry, remove the screen and wipe off any } excess phosphor from the back and sides of the screen. } } 9. Install. } } 10. A newly coated screen will outgas for a short time when it is first } placed in the microscope. Pumping times may therefore be a little longer } than normal at first. } } A very small particle size is desirable for high resolution screens and } it may be advantageous to agitate the the phosphor suspension in an } ultrasonicator before putting it into the dish. } } Hope this helps. It can get tricky and you may not get it the first time. } Patience really helps! } } Peace, } } Phil Rutledge } }
The general (daily) program for Microscopy & Microanalysis - 96 is now available on-line at the following URL.
http://www.msa.microscopy.com
This listing outlines the , daily schedule of symposia, poster sessions, platform sessions, business meetings, workshops and tutorials. A listing of authors and abstract titles will be available shortly at the same site, and will be announced when on-line.
Hi everybody, I have a question. One researcher is interested in passage of plastic microspheres about 100 nm mean diameter through the arterial wall of the experimental animal. He would like to know the path and where the microspheres are accumulating. I was asked for advice how to prepare the samples that the microspheres would be visible in TEM preparation.Plastic microspheres are translucent for electron beam and therefore invisible in TEM preparation. My question.Is there any possibility to attach an EM marker to these microspheres? Any suggestion will be appreciated. TIA John Gabrovsek CCF Cleveland, Ohio
Karen: Your problem is not new. About 30 years ago I was put in charge of an small EM lab. One of the problems which was to challenge my enthusiasm, which was rather moderated by a lack of technical acumen, was a project on the development of the cricket egg. Its leathery case withstood all ingenuity. I think that the only kind of "success" we had was with warm 2% KMnO4. This only fixes membranes and is now a fixative of ill repute.
Perhaps we should have tried 2% OsO4 or just vapour at perhaps 40 degrees C. (Take care of fumes!) Perhaps a penetrating agent like the very toxic DMSO could have helped - but I wonder: would it change structure before fixation is effective.
Glutaraldehyde is too large a molecule, although it penetrates further than Os; a tight case like the cricket's egg is more likely to be penetrated by a large atom like Os, than a large molecule. In my experience, difficult tissues like these, when surrounded GA, are more likely to go into autolysis. Fixation may only occur when the "post fixative" is added. Certainly good fixation is required prior to any plastic infiltration; it's too easy to blame infiltration when poor fixation is the more likely underlying problem.
If I had to deal with those cricket eggs now, I would look at more recently developed alternative techniques. Maybe freeze etching or perhaps freeze substitution, but I expect that some cryo fixation /sectioning techniques really holds the greatest promise.
If all of this seems difficult, try dry aleurone. That is the live, outer layer of the wheat grain. Its easy, after it is imbibed, but try it dry. "Old fashioned TEM" still has challenges greater than printing overheads and computer imaging!
Jim Darley
At 12:36 24-05-96 GMT, you wrote: } A week or so ago I presented a question how to prepare insect eggs for TEM. } I thank everyone who sent us advice in this area, however we are still } unable to successfully infiltrate the eggs. So far this is what has been done. } } 1) Standard fixation and infiltration protocol. } 2) Microwave fixation for impervious biological specimens. } 3) Pre-treatment with 1M meta periodate, extended acetone dehydration } followed by infiltration with Spurrs. } All of these methods left us with collapsed samples. } } I would like to try a pre-treatment of chitinase and possibly a combination } of chitinase and meta periodate on this sample. Does anyone out there in the } microscopy world have experience with the area or know of a reference? } } Once more } Thank you } } } ---------------------------------------------------------------------------- } --------- } Karen Vaughn Tel.(904) 392-1184 } EM Technician } University of Florida Fax.(904) 846-0251 } Electron Microscopy Core Laboratory Email. KLV-at-biotech.ufl.edu } Interdisciplinary Center for Biotechnology Research } http://www.biotech.ufl.edu/~emcl/ } 214 Bartram Hall } Gainesville, Fl 32611 } } } } } Probing & Structure Microscopy Supplies & Accessories
} Perhaps we should have tried 2% OsO4 or just vapour at perhaps 40 degrees C. } (Take care of fumes!) Perhaps a penetrating agent like the very toxic DMSO } could have helped - but I wonder: would it change structure before fixation } is effective.
I have no experience with insect larvae at all, but difficult plant material may have similar problems. Incubations with high concentrations - 10% - of DMSO in the fixative preserve plant ultrastructure very well - at least for immunofluorescence. Not sure how the material would look under the EM.
Colleagues here deal with desiccated plant material - will get back re. how they prepare material for TEM.
extra good luck,
Rosemary White __ / Department of Ecology _/ \__/ \ and Evolutionary Biology / \ Monash University / Australia \ Clayton, Victoria 3168 \ ____ / phone 61-3-9905 5670 \_/ \_*_/ fax 61-3-9905 5613 __ email rgwhite-at-sci.monash.edu.au \/ or rgwhite-at-vaxc.cc.monash.edu.au
Another fixative that may be worth trying would be acrolein ("tear gas"), which has a reputation for rapid penetration. These references may be of interest:
Luft JH, 1959. The use of acrolein as a fixative for light and electron microscopy. Anat Rec 133:305.
King JC, Lechan RM, Kugel G, Anthony ELP, 1983. AcroleIn: A fixative for immunocytochemical localization of peptides in the central nervous system. J Histochem Cytochem 31:62-68.
Grote M, Dolecek C, Vanree R, Valenta R. 1994. Immunogold electron microscopic localization of timothy grass (_Phleum pratense_) pollen major allergens PHL P I and PHL P V after anhydrous fixation in acrolein vapor. J Histochem Cytochem 42 (3 C):427-43 1.
A. Kent Christensen University of Michigan {akc-at-umich.edu}
-------------------------------------
On Mon, 27 May 1996, Probing & Structure wrote:
} Karen: Your problem is not new. About 30 years ago I was put in charge of an } small EM lab. One of the problems which was to challenge my enthusiasm, } which was rather moderated by a lack of technical acumen, was a project on } the development of the cricket egg. Its leathery case withstood all } ingenuity. I think that the only kind of "success" we had was with warm 2% } KMnO4. This only fixes membranes and is now a fixative of ill repute. } } Perhaps we should have tried 2% OsO4 or just vapour at perhaps 40 degrees C. } (Take care of fumes!) Perhaps a penetrating agent like the very toxic DMSO } could have helped - but I wonder: would it change structure before fixation } is effective. } } Glutaraldehyde is too large a molecule, although it penetrates further than } Os; a tight case like the cricket's egg is more likely to be penetrated by a } large atom like Os, than a large molecule. In my experience, difficult } tissues like these, when surrounded GA, are more likely to go into } autolysis. Fixation may only occur when the "post fixative" is added. } Certainly good fixation is required prior to any plastic infiltration; it's } too easy to blame infiltration when poor fixation is the more likely } underlying problem. } } If I had to deal with those cricket eggs now, I would look at more recently } developed alternative techniques. Maybe freeze etching or perhaps freeze } substitution, but I expect that some cryo fixation /sectioning techniques } really holds the greatest promise. } } If all of this seems difficult, try dry aleurone. That is the live, outer } layer of the wheat grain. Its easy, after it is imbibed, but try it dry. } "Old fashioned TEM" still has challenges greater than printing overheads and } computer imaging! } } Jim Darley } } At 12:36 24-05-96 GMT, you wrote: } } A week or so ago I presented a question how to prepare insect eggs for TEM. } } I thank everyone who sent us advice in this area, however we are still } } unable to successfully infiltrate the eggs. So far this is what has been done. } } } } 1) Standard fixation and infiltration protocol. } } 2) Microwave fixation for impervious biological specimens. } } 3) Pre-treatment with 1M meta periodate, extended acetone dehydration } } followed by infiltration with Spurrs. } } All of these methods left us with collapsed samples. } } } } I would like to try a pre-treatment of chitinase and possibly a combination } } of chitinase and meta periodate on this sample. Does anyone out there in the } } microscopy world have experience with the area or know of a reference? } } } } Once more } } Thank you } } } } } } ---------------------------------------------------------------------------- } } --------- } } Karen Vaughn Tel.(904) 392-1184 } } EM Technician } } University of Florida Fax.(904) 846-0251 } } Electron Microscopy Core Laboratory Email. KLV-at-biotech.ufl.edu } } Interdisciplinary Center for Biotechnology Research } } http://www.biotech.ufl.edu/~emcl/ } } 214 Bartram Hall } } Gainesville, Fl 32611 } } } } } } } } } } } Probing & Structure } Microscopy Supplies & Accessories } } Phone +61 77 740 370 Fax: +61 77 892 313 } Internet Catalogue: http://www.ultra.net/~pns/ } }
Do you fix your specimens in osmium tetraoxide ? If your answer is Yes, then it will have a better chance to locate the plastic microspheres in the arterial wall without any addition EM marker.
On Sun, 26 May 1996, John Gabrovsek wrote:
} Hi everybody, } I have a question. One researcher is interested in passage of plastic } microspheres about 100 nm mean diameter through the arterial wall of } the experimental animal. He would like to know the path and where the } microspheres are accumulating. I was asked for advice how to prepare } the samples that the microspheres would be visible in TEM } preparation.Plastic microspheres are translucent for electron beam } and therefore invisible in TEM preparation. My question.Is there any } possibility to attach an EM marker to these microspheres? } Any suggestion will be appreciated. } TIA } John Gabrovsek } CCF Cleveland, Ohio } }
[1;33m ***********************************************[0m [1;33m * Ming H. Chen, PhD *[0m [1;33m * Medicine/Dentistry Electron Microscopy Unit *[0m [1;33m * mingchen-at-gpu.srv.ualberta.ca *[0m [1;33m ***********************************************[0m
I would like to share some experience dimpling both Ti/SiC and Al/SiC. Both of these materials require mechanical thinning to less than 5 microns in order to minimize the difficulties associated with dissimilar ion milling rates between the SiC fibers and the bulk material. Aspects that need to be considered, particularly in the case of Al/SiC is that the bulk material will smear rather than grind, and the bond between the SiC fibers and the bulk material is usually weak. I have found that a minimal force applied to the specimen results in the SiC dropping out of the matrix and becoming suspended in the abrasive media. When this occurs, significant scratching of the specimen's surface will result.
After a great deal of experimentation, I was able to successfully dimple both Ti/SiC and Al/SiC. The dimpling grinder used in the research was our Model 2000 Specimen Prep System. The conditions initially utilized were a force of approximately 20 grams, a grinding rate of 1.0 microns/minute and a 3 micron diamond abrasive. Dimpling was conducted to a thickness of 15 microns when the rate was reduced to 0.5 microns/min and the abrasive was changed to 1 micron diamond. This was done until the sample thickness was 10 microns. The grinding rate was then reduced to 0.2 microns/minute, the force to 15 grams and the abrasive to 0.25 micron diamond. Dimpling was terminated at a specimen thickness of 3 microns. During the final few microns of specimen material removal the abrasive was changed every 2-3 minutes.
The key to the success in dimpling was having the ability to program a grinding rate. By establishing a grinding rate less than the material's actual removal rate (as determined by the applied force and abrasive grit size), incremental amounts of material can be readily removed. The rate control mechanism exhibits a resolution of 37 nm, and because the grinding wheel stage is supported by the rate control stage, the grinding wheel actually becomes more concentric with use. It is important to note that wheel vibration due to an eccentric wheel is the single most contributing factor to specimen breakage.
For a more complete description please refer to my paper in MRS 199.
I hope this helps.
Paul
paul.fischione-at-internetmci.com Paul E. Fischione, President E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 USA Phone (412)325-5444 FAX (412)325-5443
------- FORWARD, Original message follows -------
Dear lists:
My colleague would like to pose a question into this cyber space:
} Hello all, } } Does anyone know the spectroscopic properties(peak emission wavelength and } FWHM, etc.) of the photochromic pigment such as below? } } 1) Name of the photochromic pigment: } 1,3,3-trimethylindolime-6-nitrobenzospiropyran, generally called TNSB for } short. } 2) Circumstance: } This photochromic pigment is in the Silicon oil. } 3) Excitation source: } The excitation source is the Nitrogen Laser(wavelength is 337.1nm). } } Many thanks in advance. } ------------------------------
An excellent, short handbook about EDX can be found in the Royal microscopical Society series #05: "X-ray Microanalysis in Electron Microscopy for Biologists" by A.J. Morgan. It is about 70 pages long and glosses over introductions and terms, x-ray production, x-ray detection, qualitative analysis, quantitative analysis, and specimen preparation.
Oxford University Press ISBN 0-19-856409-0
Dennis Shubitowski University of Michigan School of Dentistry dshubito-at-umich.edu
Dennis Shubitowski University of Michigan School of Dentistry dshubito-at-umich.edu
If anyone has experience problems with the light fixture located on the head of an LKB Ultrotome Nova and has fixed it or found the problem, please send me your feedback, it would be greatly appreciated.
Thank You,
Philip _______________________________________________ SCOTT SCIENTIFIC P.O. Box 66552 Station Cavendish Montreal, Quebec, H4W 3J6, Canada
We have made apertures for many years. More information on our apertures is available on the WWW at http://www.fullam.com/aperture.htm or you can contact us at the numbers listed in our signature.
Dianne Fullam
} Does anyone know where I can purchase TEM apertures (condenser, objective, } and selected area) for a Philips EM301 (besides Philips). } } Thanks } } ************************************************************************** } Lucille A. Giannuzzi, Ph.D. phone: 407 823-5770 } University of Central Florida fax: 407 823-0208 } Materials Science Program email: lag-at-pegasus.cc.ucf.edu } Dept. of Mechanical and Aerospace Eng. } Orlando, FL 32816-2450 } ************************************************************************** } } } Ernest F. Fullam, Inc. Phone: (518) 785-5533 FAX: (518) 785-8647 E-Mail: pdf-at-fullam.com
} To: John Gabrovsek {gabrovj-at-cesmtp.ccf.org} } From: gwe-at-biotech.ufl.edu (Greg Erdos) } Subject: Re: TEM em-markers } Cc: } Bcc: } X-Attachments: } } } Hi everybody, } } I have a question. One researcher is interested in passage of plastic } } microspheres about 100 nm................ } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } Your researcher might consider magnetic microspheres, since they are loaded with iron and are quite visible at the TEM. I have embedded and section them using routine techniques. Also most of these plastic microsperes have reactive surfaces so you might be able to attach a protein that has been coupled to small colloidal gold } ******************************************************* Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
Sorry to bother, but could someone please let me know how a new person would sign on to this list server? It's been so long that I have been on that I have forgotten and my recent hard disk crash wiped out the old messages for instructions.
TIA
Jim
***************************************** Jim Crandall, Ph.D. Senior Neurobiologist Department of Developmental Neurobiology E. Kennedy Shriver Center 200 Trapelo Road Waltham, MA 02154 USA
tel 617-642-0278 fax 617-893-4018 email jcrandall-at-shriver.org *****************************************
I've always ordered apertures (platinum and gold foil) for my Philips 301 through Electron Microscopy Sciences. Their phone # is 1-800-523-5874. I've been happy with their products and service, and their prices are very competitive.
Debbie Cassout E.M. Dept. Texas Veterinary Medical Diagnostic Lab. College Station, TX
} } } Lucille A. Giannuzzi {lag-at-pegasus.cc.ucf.edu} 05/24/96 10:43am } } } Does anyone know where I can purchase TEM apertures (condenser, objective, and selected area) for a Philips EM301 (besides Philips).
Thanks
************************************************************************** Lucille A. Giannuzzi, Ph.D. phone: 407 823-5770 University of Central Florida fax: 407 823-0208 Materials Science Program email: lag-at-pegasus.cc.ucf.edu Dept. of Mechanical and Aerospace Eng. Orlando, FL 32816-2450 **************************************************************************
Greetings: Has anyone had experience with Feulgen staining of LR Gold or LR White sections? We would like to use this procedure to verify fertilization of frog oocytes. Methods, tips, pitfalls, etc. would be appreciated.
Thanks in advance,
Arthur R. Hand Central EM Facility UConn Health Center Farmington, CT
My interest in the CIC Services took me to their home page where I could e-mail them for more information. I think the idea for this type of "insurance company" pooling of service contracts makes sense. In fact that is exactly what each individual service contract really amounts to... insurance to repair future problems. Their response to my e-mail is attached for anyone interested. Look into it or not as you wish. Linda Fox lfox1-at-wpo.it.luc.edu Received: from quickmail.cicagency.com ([206.104.48.12]) by mail.cicagency.com (Netscape Mail Server v1.1) with SMTP id AAA95 for {lfox1-at-wpo.it.luc.edu} ; Mon, 27 May 1996 15:06:47 -0500 Message-ID: {n1378913100.90616-at-quickmail.cicagency.com}
Subject: Time: 2:44 PM CIC Services Date: 5/27/96
Thank you for the notification of our services being discussed on your list. You are exactly right in your assessment that our services do not preclude anyone from using the original manufacturer as a service provider. In fact, under CIC's program, most facilities continue to use the service provider they are most comfortable with, which is a popular feature of CIC's asset management services.
In summary, the program works as follows:
CIC takes your existing service and maintenance contract(s) and combines them into one comprehensive asset management agreement. In the process, you save 15 to 25% over manufacturer service contracts. When a service event occurs, you call the service vendor OF YOUR CHOICE, who comes out and effects the repair as usual. Upon receipt of the invoice, you simply submit it to CIC who pays the service vendor directly, or reimburses you after you pay it, depending on the terms of our agreement. In exchange, you pay CIC a fixed, budgetable amount that is (again) 15 to 25% below what your facility was paying.
I am unsure as to whether we would be allowed to subscribe to your list, as many lists preclude private companies from subscribing. I would be greatly appreciative if any misconceptions about our company and the service we provide could be corrected. We are very proud of the efficiency and cost-effectiveness we bring to facilities with high-tech equipment, and we are also proud of the many endorsements we hold from major universities and hospitals across the U.S. and Canada. Currently, we provide such services to the University of Florida, the University of Ohio, Texas A&M, Cornell, Colorado State, Louisiana State, UC-Davis, the University of Georgia, and Virginia Tech. In addition, there are hundreds of hospitals in our client base as well.
Perhaps you could copy my message to your list? Either way, thank you for your inquiry and I will pass along your questions to a CIC Regional Manager in your area. Thanks again.
Hi all, I am interested in getting in touch with other people doing microscopy work (research or routine) of silicon and possibly other semiconductors, for sharing of experiences, and possible collaboration.
I am currently working on: better understanding the role of bulk microdefectsa in the gettering properties of silicon. (TEM, Optical, lifetime , scanning Infra-Red Microscopes)
fundamental understanding of crystal growth - vacancy and interstitial incorporation, grown in defects etc.. (tem, optical etc..)
SOI - interface, diffusion through etc.. (TEM)
developement on IR scanning tools for use in semiconductor bulk defect characterization.
I am also interested in a variety of other areas of semiconductor research, including problems related to device growth (metallization, oxide-related, implantation etc..).
I work at MEMC Electronic Materials Inc, which is one of the world's largest silicon wafer maker. Thus obtaining material (and funding if the problem is relevant) is usually not a problem.
cheers
Lucio
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ~~~~~~~~~~~~~~~~~~~~ Lucio Mule'Stagno MEMC Electronic Materials Inc University of Missouri -St.Louis Silicon Materials Research Group Physics Dept., 501 Pearl Dr., 8001 Natural Bridge rd., St.Peters, St.Louis MO 63376 MO 63121 tel: 314 279 5338 314 516 5931/3 fax: 314 279 5363 314 516 6152 email: lmulestagno-at-memc.com luciom-at-newton.umsl.edu URL:http://www.newton.umsl.edu ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ~~~~~~~~~~~~~~~~~~~~
Message-ID: {n1378843917.3500-at-quickmail.llnl.gov} "South Bay Technology" {73531.1344-at-CompuServe.COM} X-Mailer: Mail*Link SMTP-QM 3.0.2
RE} TEM: Dimpling Ti with SiC fibers 5/28/96
A number of years ago we discovered that CBN (cubic-Boron Nitride) will dimple metals such as Al, Ti, Zr, Ta, Pt, Cu, Ni,...etc. much faster, with less damage and smearing than diamond. This is for two possible reasons. Diamond can chemically react with the metals and become bonded to the sample, thus preventing any further dimpling, in fact you may find that your dimpling wheel (steel, phosphor bronze, etc.) will actually wear away faster. Second, as the abrasive action is taking place during the dimpling the diamond powders become covered with the removed sample material and you are no longer dimpling with diamond but with particle coated with metal-hence smearing and gauling. CBN particles have different shaped corners and edges and will not react with the metals. It is still necessary to change slurries more often when dimpling metals and the process is slower that say dimpling Silicon.
CBN of course will not dimple SiC very well. The trick is to mix the CBN with the diamond in the relative proportion of the sample composition. We have found it possible to dimple two phase materials of vastly different hardness at nearly identical rates using this multi-abrasive slurrey technique. We start out with 2-4um then finish with 0-2um. We do not polish with a soft cloth, this will instantly cause relief between the soft and hard phase.
Good luck
Mark Wall Lawrence Livermore National Lab 510-423-7162
--------------------------------------
You've been so helpful in the past that I thought I'd give it another go!
I am looking for any help I can get on dimpling Ti with SiC fibers. I don't have much more information than that at this point as I am relaying this message for someone else. If anyone can help, I'd really appreciate it.
While dimpling was the question, if you have information on Tripod Polishing, ion milling, or any other technique that would also be quite useful.
Thank you very much!
Best regards-
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: 73531.1344-at-compuserve.com sbt-at-msa.microscopy.com http://www.southbaytech.com
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy.
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Grateful Med (NLM) computer reference search program for the Mac V2.2. I just upgraded to Mac system 7.5.3 which comes with Open transport 1.1. In trying to start GM 2.2 there is a crash every time. I tried everything that makes sense to solve the problem and communicated the techn services at the NLM with no results. Anyone with similar problems and possible fixes please contact me direclty.
********************************************************************* *Cesar D. Fermin, Ph.D \|T|/ Fax (504) 587-7389 * *Tulane Medical School /|M|\ Voice Mail (504) 584-2618 * *Pathology/SL79 \|C|/ Secretary (504) 584-2436 * *New Orleans La 70112-2699 /|*|\ Lab/Techn. (504) 584-2521 * * {Fermin-at-tmc.tulane.edu} ----} Prof. Pathology & Otolaryngology * *http://www1.omi.tulane.edu/departments/pathology/fermin/cdftop.html* *********************************************************************
Does anyone know of a blocking procedure for elastin in frozen thick cryostat sections. Specifically we're using anti-rabbit FITC and are getting non-specific staining of elastin, which we would like to block. The tissue (rat tail artery and mesenteric artery) was fixed in 4% parformaldehyde and embedded in Tissuetek. This is'nt directly an EM question, but I thought someone might know anyway.........
I have been asked to do immuno on bacteria (E. Coli). Has anyone had any experience with this ? What fixative is the best to use ? Should I do pre or post labeling ? I have tried two fixatives with this bacteria and 2% Glut worked better than 0.5% Glut and 4% Para. Anybody have any suggestions ? Thank you !!
Barbara Hartman Schering-Plough Research Institute 201-579-4343 201-579-4211 (FAX)
Message-Id: {199605291346.JAA15191-at-vaxserv} X-Sender: nnicklaus-at-cave.sarnoff.com X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Does anyone know of "attachments" for Zeiss microscopes which add confocal laser scanning abilities?
I have both upright and inverted Zeiss microscopes. Both use the infinity corrected, ICS, optics. I want to be able to use these objectives amongst my various microscopes.
P.S. This question will be placed with both the confocal microscopy group and the microscopy group.
P.P.S. I had previously asked about video rate confocal attachments. This inquiry is for a different application (surface studies in reflection mode).
Message-Id: {199605291352.JAA15206-at-vaxserv} X-Sender: nnicklaus-at-cave.sarnoff.com X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Does anyone know of "attachments" for Zeiss microscopes which add confocal laser scanning abilities?
I have both upright and inverted Zeiss microscopes. Both use the infinity corrected, ICS, optics. I want to be able to use these objectives amongst my various microscopes.
P.S. This question will be placed with both the confocal microscopy group and the microscopy group.
P.P.S. I had previously asked about video rate confocal attachments. This inquiry is for a different application (surface studies in reflection mode).
P.P.P.S.: 2nd time I sent this to Microscopy Group - forgot to add subject to the first transmission.
Would appreciate any help with the following: Am trying space group determination for the first time. If I understand correctly, which is questionable, for +G and -G conditions you move the condenser aperture to the Bragg condition which would be halfway between the bright field and the dark field disc at the zone axis orientation. In one direction is OK, with little leeway, in the opposite direction a severe cut-off occurs on all discs, say 50-70%. It appears that this is a fixed aperture or polepiece (CM20). So where have I stumbled? (Condenser aperture size 200 micrometres) Thanks Mike Witcomb
Dr MJ Witcomb Electron Microscope Unit University of the Witwatersrand Private Bag 3 WITS 2050 South Africa
} } Your researcher might consider magnetic microspheres, since they are loaded } with iron and are quite visible at the TEM. I have embedded and section } them using routine techniques. Also most of these plastic microsperes have } reactive surfaces so you might be able to attach a protein that has been } coupled to small colloidal gold } } } Greg Erdos
I would go with the latter recommendation, as opposed to iron or osmium (or other heavy metal) loaded microspheres. Aside from possible reactivity/bodily reaction problems due to the heavy metals, the metals will chang the specific gravity, and therefore flow characteristics of the spheres. This would change their distribution within the blood vessels, and so possibly affect where, how, and if they pass out of the vessels into the surrounding tissues.
&&&&&&&&&&&& Illigitmi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&
Philip Oshel soon-to-be-closed Center for Electron Microscopy University of Illinois Rm 74 Bevier Hall 905 S. Goodwin Ave. Urbana, IL 61801 (217) 244-3145 oshel-at-ux1.cso.uiuc.edu
************ looking for a job again ****************************************
A standard fixation technique in the drosophila world requires several steps: 1) dissolve the chorion with ~5% Na hypochlorite (bleach) ~5 minutes then rinse usually with PBS and .3% Trition X-100; 2) fix in a solution of heptane and your choice of buffered aldehyde;3) pipet the samples off the interface and transfer to a heptane:methanol and shake vigorously--the vitelline membrane will burst when the tissue swells. 4) collect the samples that sink to the bottom and rehydrate in beffer. This technique is called phase-partition fixation. See M. Zalokar & I Erk, "Phase-Partition Fixation and Staining of Drosophila Eggs", Stain Technology, Vol 52, No. 2, 1977, pp. 89-95.
Note the above is for embryos--it may not work with larvae that have a more developed membrane. I usually slice very small larvae in half or at the tip while submerged in fix or if they are larger I pin them out and dissect them as appropriate. This requires a very sharp blade and good microdissection technique--most grad students and post-docs can do it. Contact me directly for more tips and hints.
Larry Ackerman mishot-at-itsa.ucsf.edu The Laboratories of Lily & Yuh Nung Jan Voice (415) 476-8751 Howard Hughes Medical Institute UCSF, Box 0724, Rm U426 FAX (415) 476-5774 533 Parnassus Ave. San Francisco, CA 94143
Greetings, Does anyone know if it's true that chloral hydrate actively quenches fluorescence??? We found that some whole mount preps that had been cleared in chloral hydrate were non-fluorescent, and I have heard vaguely about quenching by chloral h. I was wondering if anyone had any similar experience, or understood why from first principles such a molecule would be expected to quench. Or are there folks out there who get great fluorescence from specimens cleared in chloral h. Many thanks, Tobias Baskin
There has been many comments and questions about CIC service.
My comment/question is since you would no longer have a service contract with the manufacture, is the same response time available?
Most companies put the service contract customers first, hence quick response time. Since this would no longer be the case, the question now is how long can one wait for service. I have a feeling it may be quite some time, maybe even weeks. A long wait can be very costly in down time. So in reality I think there would not be a savings, more like a loss.
Eric is the champion. His suggestion fixed the problem. Solution, removed the open transport that comes with the 7.5.3 upgrade and do a custom install as described by Eric. What a relief! T H A N K S E R I C! ! ! ! } Grateful Med (NLM) computer reference search program for the Mac V2.2. } I just upgraded to Mac system 7.5.3 which comes with Open transport 1.1. In } trying to start GM 2.2 there is a crash every time.
You are not going to like hearing this but you will have to remove the OT1.1 software and step back to the normal communications extensions you were using pre-7.5.3. Open Transport has a mass of new extensions that are not compatible with the majority of communications packages currently available.
--------------------------------------------------------------- Eric Chapman - Eagle River Alaska _/_/_/ _/_/_/ _/_/ Hemodialysis Patient _/ _/ _/ _/ _/ _/ _/_/ _/ Sic Gorgiamus Alles _/ _/ _/ _/ _/ Subjectamos Nunc _/_/ _/_/_/ _/_/ --------------------------------------------------------------- The easiest method is to go into your system folder and delete all open trasport extensions. Then, use a pre-7.5.3 install disk or CD and use custom install. Just install the communications toolbox extensions and you should be in business. We have a few of the new 7500 boxes in our shop here and I will run over and talk to them about the problems we have been having with open transport. I will get back to you before the end of the day.
A complete listing of all Authors and Manuscript Titles for Papers and Posters which will be presented at the Microscopy & Microanalysis-96 Meeting in Minneapolis, Minnesota August 11-15, 1996 is now on-line at the URL
http://www.msa.microscopy.com
Look for the hot link entitled "Author/Title Index" under the Annual Meetings Area.
See you in Minneapolis...........
Nestor Your Friendly Neighborhood SysOp (& Program Chairman- Microscopy & Microanalysis-96)
Try the Zeiss people at their US headquarters: 800-356-1090. You may be able to obtain more information from their Web site. Access it through ours: http://www.shore.net/~catalogs. You'll find many products featured there. If you call, please tell them we sent you.
Best of luck,
Elinor Solit, The Microscope Book
On Wed, 29 May 1996, Francisco J. Hernandez wrote:
} I'm searching for used phase contrast microscopes to } buy. I will be very grateful for any information. } } Francisco J. Hernandez-Blazquez } fjhblazq-at-spider.usp.br } fjhblasq-at-biomed.icb2.usp.br } } }
-- [ From: Garber, Charles A. * EMC.Ver #2.10P ] --
The American Association for Laboratory Accreditation (A2LA) has announced a new listserver for the discussion of issues relating to laboratory accreditation. Specifically:
"Who is doing what, how and why organizations are becoming accredited, what the successes and concerns have been, what concepts, methods, software and organizatons have been beneficial, etc"
Subscription information can be found at the following URL:
http://www.fasor.com/~iso25/listserv.html
The A2LA home page is accessible from the above URL code. If anyone thinks laboratories are not being accredited, they would have to be impressed with the long listing of laboratories under current accrreditation by A2LA. More than a few EM oriented laboratories are covered by the A2LA accreditation program, including our own laboratory. What ever money we have had to spend to maintain our accreditation program, we are completely convinced comes back to us in the form of significantly lower costs associated with "rework" (e. g. having to do work over again because it was not done right the first time).
Chuck
Hello Charlie:
I am sending this non-list server comment to you, off the listsever in order that my comments not be misinterpreted by anyone since we are one of the several main sellers of osmium tetroxide to the world wide EM community.
At the risk of appearing to be not so knowledgeable, let me give you my own thoughts on this issue:
a) There is no reason why freezing aqueous osmium tetroxide should in any way, shape, or form degrade the "quality" of the material in the vial. Therefore it would not surprise me that no one reported any erosion of quality.
b) My biggest fear has been from both a safety and also product liability standpoint. The safety aspect comes into play because as the aqueous component drops through 4 deg. C, where there is a maximum in the expansion curve, it just seemed to me to be the classic accident waiting to happen: Any ampoule with glass containing even the slightest defect (would be a major stress riser), and with the glass being so brittle, I would expect that the ampoule would spontaneously shatter.
I just can't imagine a worse kind of clean up problem and depending on when it happened and when it was discovered, who knows what other kind of damage could occur. Or what kind of injury could occur. Of course, there was one posting that indeed did report a broken ampoule in the freezer, the failure being presumably due to the freezing.
From a product liability standpoint, I would be very reluctant to recommend this as a method of storage. I admit that this is "defensive" business practice, but I sure don't want to have to start hiring lawyers to defend our firm against legal action, in the event we should have suggested freezing as an acceptable method of storage and have it then result in an accident and damage, worse yet, some kind of human injury.
If someone really did want to store the osmium tetroxide (aqueous) in a freezer, then there are plastic sleeves that I would recommend they be place in, if for no other reason that in the event there should be some kind of a failure of the glass ampoule, then the plastic sleeve would at least tend to contain any osmium from getting out into the general part of the refrigerator.
Now for what might at least have the appearance of a commercial pitch, and I don't mean it to be such a "pitch", we have been offering aqueous osmium tetroxide for more than twenty years. If one practices ultra clean practices, both in terms of the glass ampoule before filling and also with the water diluent (all the osmium starts out with the same starting purity), and also the use of a high quality dry nitrogen for filling the head space, then the product should last virtually indefinitely without refrigeration! Yes, that is right. Now not all people offering aqueous osmium tetroxide are ampuoling it to that standard as evidenced by the fact that people have had problems and have resorted to freezing it in order to extend the shelf life. I am almost certain that our aqueous osmium tetroxide is not the cheapest around, but people who have been using it, just don't have to go to such lengths in order to keep it active and in the long term, the higher stability is well worth any few pennies more in the selling price.
My own gut feeling is that the typical end user should just be purchasing aqueous osmium tetroxide of the highest quality and avoid altogether the safety and other risks associated with the freezing in a glass ampoule this really hazardous material.
You have my permission to re-post this message on the listserver, if you think it would be of more general interest. You also have my permission to edit it as you see fit.
Best regards.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service:spi2spi-at-2spi.com
Dear all, I realise that this question has been asked in the recent past but I didn't think to keep the responses.
What are peoples experiences with the use of high res laser printers or other types of printer for printing EM images? Do you have any printer that you would recommend? I would be interested in using it for printing out a wide variety of TEM and SEM images of materials science specimens.
I would also be very glad if anyone has a summary of previous messages that have been posted on this subject that they could forward to me.
Thanks
_________________________________________________________________ Ian MacLaren, Telephone: 0121 414 3447 IRC in Materials, FAX: 0121 414 3441 The University of Birmingham, email: I.MacLaren-at-bham.ac.uk Birmingham B15 2TT, England. _________________________________________________________________
In everyone’s opinion, what is the best commercial Image processing and analysis package available on the market today. One that offers excellent support, intuitive ease of use, fully editable scripts or macros, and can handle a wide variety of applications. Currently, I am using Kontrons IBAS v2.0, thinking about upgrading to a new system. Thanks Gregory Argentieri Sandoz Pharmaceuticals Corp 201-503-8617 Fax 201-503-6339
} We currently have users book microscope time by writing with a pen on } booking sheets made of paper. We are contemplating changing this antiquated } system to a computer booking system which would permit people to book (and } to check existing bookings) over the net. } } Does anyone know of software to do this? Please take into account that we } would want the software to meet the following conditions: } } It should work for several instruments. } It should prevent one user erasing another's booking. } It should allow staff to change the bookings of users. } If a user deletes his or her own booking, a record of the booking and the } time of the cancellation should be kept. } It should be possible to program booking rules into the system. These rules } may be different for each instrument. The rules would include such things } as how many sessions may be booked at any one time and how long a session } may be booked. } It should be possible to include various degrees of access for the } users. Some users may be allowed to book evenings and weekends while others } may only book daytime sessions. } } We feel that unless the software we use can meet most of these requirements, } we will be better off staying with pen and paper. Your suggestions would be } most welcome. } Alwyn Eades eades-at-uimrl7.mrl.uiuc.edu } } Center for Microanalysis of Materials, University of Illinois, } 104 S. Goodwin, Urbana, Illinois, 61801-2985, USA } 217-333-8396, Fax 217-244-2278.
You should be able to do all of this by networking one or more database files by using a database manager such as Filemaker Pro (Claris).
Specifically: } It should work for several instruments. you could use several different files. } It should prevent one user erasing another's booking. by using access privileges and pass words you can restrict what particular users could see or do in different layouts } It should allow staff to change the bookings of users. this could be allowed by their password and access privileges } If a user deletes his or her own booking, a record of the booking and the } time of the cancellation should be kept. A enw current record could be made and the old one kept. } It should be possible to program booking rules into the system. These rules } may be different for each instrument. The rules would include such things } as how many sessions may be booked at any one time and how long a session } may be booked. different files with different sets of access privileges. } It should be possible to include various degrees of access for the } users. Some users may be allowed to book evenings and weekends while others } may only book daytime sessions. different access privileges for different sets of users.
The drawback to this approach is that a copy of the program (or site license) all the user computers (or at least for all of those running at once) should be purchased in order to be legal. Unless your campus has a license. On the other hand database managers can be used to maintain several different files.
Bill
Bill Trevarrow Institute of Neuroscience University of Oregon 1254 Eugene, OR 97403-1254 Off.Tel: (541) 346-4598 Fac. Tel: (541) 346-4512 Fax: (541) 346-4548 e-mail: trevarro-at-uoneuro.uoregon.edu
} In everyone=92s opinion, what is the best commercial Image processing and } analysis package available on the market today.=20 } One that offers excellent support, intuitive ease of use, fully editable } scripts or macros, and can handle a wide variety of applications.=20 } Currently, I am using Kontrons IBAS v2.0, thinking about upgrading to a n= ew } system. } Thanks } Gregory Argentieri } Sandoz Pharmaceuticals Corp } 201-503-8617 } Fax 201-503-6339 } =20 } =20
That's easy -
For 2D - MetaMorph by Universal Imaging For 3D - VoxelView by Vital Imaging
Michael
Dept. of Biology Univ.of New Mexico Albuquerque, NM 87131
} In everyones opinion, what is the best commercial Image processing and } analysis package available on the market today. } One that offers excellent support, intuitive ease of use, fully editable } scripts or macros, and can handle a wide variety of applications. } Currently, I am using Kontrons IBAS v2.0, thinking about upgrading to a new } system. } Thanks } Gregory Argentieri } Sandoz Pharmaceuticals Corp } 201-503-8617 } Fax 201-503-6339 } }
Personally, I have had a very good experience with the new Quantimet Q600 system from Leica. It is a fast traditional image analysis system (like the IBAS) but is user friendly and the macros can be written with the mouse (very simple). In the high resolution mode it can handle images above 1K X 1K pixel.
My two cents,
Lars Bjork Dept Immunology Wenner-Gren Institute Stockholm University SWEDEN
Helpful people: I'm having trouble with some seed material...we need to look at the pericarp(s) of these guys through the seeds' development. The early stages were cake, but I'm having trouble with older material - it is more "woody" as we go. I've found several seed prep protocols, but I was hoping someone out there has a tried and true method... Thanks! Tamara Howard CSHL
Here's another suggestion I'd like to in the mix. I worked with a materials person a few years ago doing some TEM on mixtures of polymeric materials. The one I remember was a mixture of Polysytrene and Starch. I thought we were going to have real problems visually separating the two materials in the TEM thin sections, and was ready to play all sorts of staining games. But he took the first step and simply sectioned the material (unfixed, unembedded) and popped them into the scope and we had absolutely no problem seeing them in the scope! One phase was nearly electron transparent (light grey) and one phase was nearly electron opaque (dark grey - sorry I do not remember which one was which).
Before trying weird stains and possible immuno labeling horrors: Have you tried emmbeding the spheres (or serval possible different material types) sectioning them and looking to see what shows up ijn the TEM? I would think the nice, neat, very uniform density circles should be fairly easy to find in amongst the 'mess' of biological material in the scope.
(I am a Biologist - primarily - and any one offended by refering to a 'mess of biological material' needs to relax and get their sense of hummor back.)
Richard E. Edelmann Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
Message-Id: {2.2.32.19960531132011.00751be8-at-noesisvision.com} X-Sender: ln-at-noesisvision.com X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable
At 08:20 PM 5/30/96 -0400, you wrote: } In everyone=92s opinion, what is the best commercial Image processing and } analysis package available on the market today.=20 } One that offers excellent support, intuitive ease of use, fully editable } scripts or macros, and can handle a wide variety of applications.=20 } Currently, I am using Kontrons IBAS v2.0, thinking about upgrading to a new } system. } Thanks } Gregory Argentieri } Sandoz Pharmaceuticals Corp } 201-503-8617 } Fax 201-503-6339 } }
You can try Visilog from Noesis Vision Inc. We have been developing software for over 11 years and are in the process of introducing a brand new 32 bit version compatible with Windows 95 and NT. Visilog runs on both PC and Unix workstations and provides a wide selection of image processing algorithms, macro language(editable), a redone easy to use GUI (user interface)and drivers for various grabbers and stages. Visilog is widely used in the microscopy field.
= =20 ---------------------------------------------------------- --------------------------------------- Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com http://www.cam.org/~noesis 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada
Join the Noesis NewsGroup at Visilog-at-noesisvision.com ---------------------------------------------------------------------------- ---------------------
At 04:41 PM 5/30/96, you wrote: } Dear all, } I realise that this question has been asked in the recent past but I didn't } think to keep the responses. } } What are peoples experiences with the use of high res laser printers or } other types of printer for printing EM images? Do you have any printer } that you would recommend? I would be interested in using it for printing } out a wide variety of TEM and SEM images of materials science specimens. } } I would also be very glad if anyone has a summary of previous messages that } have been posted on this subject that they could forward to me. } } Thanks } } _________________________________________________________________ } Ian MacLaren, Telephone: 0121 414 3447 } IRC in Materials, FAX: 0121 414 3441 } The University of Birmingham, email: I.MacLaren-at-bham.ac.uk } Birmingham B15 2TT, England. } _________________________________________________________________ } }
Hi Ian. If you have www access, then I the man you want to hear from. Go to the address listed at the bottom of this message and look in the "Tips & Tricks" section. There you will find a link to a set of links to a set of links... Look for the "Image processing, conversion, and software" link and you will find a recent discussion that came across the list. If for some reason you do not have web access, get it , or e-mail me back and I will get the file to you. As an answer, we have a Lexmark Optra R, The cheap, get them proofs, 1200 dpi laser and the Kodak xls 8600 dye sub, bet you can't tell which is the photograph printers. Both have been excellent and just recently withstood the battering only a group of inexperienced students could dish out flawlessly. In all we have been pleased. Hope this helps.
Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 904-392-1295 ICBR EM Core Lab fax 904-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/
Responding to the message of {v01510103add3f6f4208b-at-[128.223.140.24]} from trevarro-at-uoneuro.uoregon.edu: } } } We currently have users book microscope time by writing with a pen on } } booking sheets made of paper. We are contemplating changing this antiquated } } system to a computer booking system which would permit people to book (and } } to check existing bookings) over the net. } } } } Does anyone know of software to do this? Please take into account that we } } would want the software to meet the following conditions: } }
} You should be able to do all of this by networking one or more database } files by using a database manager such as Filemaker Pro (Claris). } } Specifically: } } } It should prevent one user erasing another's booking. } by using access privileges and pass words you can restrict what particular } users could see or do in different layouts
I have found it very difficult to get Filemaker Pro to do something like this. We have attempted to get around this problem by having a Web browser front end that communicates with Filemaker Pro using an applescript cgi which does the access privelege part.
The disadvantage of the system is that, even for a University, the number of copies of Filemaker we have (one for each microscope for logging of hours rather than booking of time) represents a ridiculous expense. We could have a Web browser do the logging part too, but suspect that it will get used for Web browsing rather than data entry!
The main advantage of the system is that relatively little human intervention is necessary for the generation of monthly billing statements.
Stuart McKernan stuartm-at-maroon.tc.umn.edu CIE Microscopy Center, University of Minnesota Office: (612) 626-7942 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
alwyn eades wrote: } } We currently have users book microscope time by writing with a pen on } booking sheets made of paper. We are contemplating changing this antiquated } system to a computer booking system which would permit people to book (and } to check existing bookings) over the net. } } Does anyone know of software to do this? Please take into account that we } would want the software to meet the following conditions: } } It should work for several instruments. } It should prevent one user erasing another's booking. } It should allow staff to change the bookings of users. } If a user deletes his or her own booking, a record of the booking and the } time of the cancellation should be kept. } It should be possible to program booking rules into the system. These rules } may be different for each instrument. The rules would include such things } as how many sessions may be booked at any one time and how long a session } may be booked. } It should be possible to include various degrees of access for the } users. Some users may be allowed to book evenings and weekends while others } may only book daytime sessions. } } We feel that unless the software we use can meet most of these requirements, } we will be better off staying with pen and paper. Your suggestions would be } most welcome. } Alwyn Eades eades-at-uimrl7.mrl.uiuc.edu } } Center for Microanalysis of Materials, University of Illinois, } 104 S. Goodwin, Urbana, Illinois, 61801-2985, USA } 217-333-8396, Fax 217-244-2278.
Alwyn,
Try FileMaker Pro by Claris. It is available for Windows, Windows95 and Macintosh. It is a fully relational, customizable, open architecture database that has extensive password level protection. It can be learned in a few hours and has endless capabilities. Trying to find a specific software for "microscope sign-up" will be close to impossible unless one of these kind folks on this server will share one of their previous efforts. Even then you will find it is a template from a relational database or spreadsheet.
Good Luck,
Lawrence Kordon Nikon, Inc. Columbia, Maryland nikon-at-jagunet.com
} Date: Fri, 31 May 1996 14:59:10 -0500 } To:Kathy427-at-aol.com } From:loakford-at-hsc.unt.edu (Larry Oakford) } Subject:Re: Comment/Question about Service Contracts } Cc: } } I concur wholeheartedly with Kathy's comments below. My } understanding is that all non-service contract and non-installation work } is last in line to get service from all major electron microscope service } providers. It also means that if a contract call comes in while they are } at your site they may have to leave and attend to it before getting back } to your problem. I have known of non-service contract labs having to wait } as long as 3 months to see a service engineer, and this was from a major } TEM manufacturer. If possible extended down time is nondisruptive to the } functioning of your lab then I would say go ahead with the CIC plan, } otherwise fight with every breath in your body to maintain a standard } service contract, even a modified one with less coverage or extended } response times, most service organizations are willing to negotiate. You } may get lucky and have a microscope that stays healthy and never needs to } have a service call, but you may be like the individual I know who had to } wait 3 months to get operational again while fending off users who needed } to use the microscope. There is a lot of truth in the saying "you get } what ya pays for". I hope this helps. } } P.S. It is also true, as pointed out by another writer, that if you decide } to return to a service contract after the present one runs out, and you } did not renew, it will cost you. You will need to go through a } pre-service contract routine where all expenses(time, travel & parts) are } directly charged above the cost of the new service contract (this can } amount to an extra bill of $2000 or more). This is a standard practice in } the microscope service industry concerning lapsed contracts, both } manufacturers and independent providers alike. } } } There has been many comments and questions about CIC service. } } } } My comment/question is since you would no longer have a service contract with } } the manufacture, is the same response time available? } } } } Most companies put the service contract customers first, hence quick response } } time. Since this would no longer be the case, the question now is how long } } can one wait for service. I have a feeling it may be quite some time, maybe } } even weeks. A long wait can be very costly in down time. So in reality I } } think there would not be a savings, more like a loss. } } } } Kathy }
In everyone’s opinion, what is the best commercial Image processing and analysis package available on the market today. One that offers excellent support, intuitive ease of use, fully editable scripts or macros, and can handle a wide variety of applications. Currently, I am using Kontrons IBAS v2.0, thinking about upgrading to a new system. Thanks Gregory Argentieri Sandoz Pharmaceuticals Corp 201-503-8617 Fax 201-503-6339