John: We do around 500 renal biopsies per year and all the sections are mounted on 200 mesh uncoated copper grids. We have an 8 year old Hitachi 7100 and use 60kv. The majority of the glomerulus can be viewed with the 3-4 serial sections lying randomly across the grid bars. We do not need a picture of the whole glomerulus, rather most pictures are between 3,000 and 10,000X. Dr. Tibor Nadasdy is the renal pathologist and decided last year that all our renal biopsies would be captured with the digital camera onto a computer and sent up to him via a network to his computer. So, at the present time we use very little EM film. He diagnoses each biopsy and e-mails representative digitized images to the nephrologists.
Karen L. Jensen, M.S. Project Manager & Associate Scientist Electron Microscopy Research Core
-----Original Message----- } From: "JHoffpa464-at-aol.com"-at-sparc5.microscopy.com [mailto:"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com] Sent: Friday, March 30, 2001 2:20 PM To: microscopy-at-sparc5.microscopy.com
Hello -
Has anyone out there tried to adhere xylem sections to pre-coated microscope slides like Fisher Probe-On Plus slides? I've tried and had a zero percent section retention. I can imagine that this is because of the scarcity of live cells (and plasma membranes). I've tried slow air drying and various temperatures on the slide warmer. The sections are 15 microns, and are from fixed (buffered paraformaldehyde) but not embedded samples. I'm about to try Poly-L-lysine and amino-acyl silane treated slides, but I'm not too hopeful because they (at least Poly-L) rely on the same positively-charged surface principle. I want to avoid gelatin or albumin subbing because I'm treating sections with protease, and also want to minimize background staining. Any tips would be greatly appreciated.
Rachel
****************************************** Rachel Spicer Biological Laboratories 3119 Organismic and Evolutionary Biology Harvard University 16 Divinity Avenue Cambridge, MA 02138
Rishi Raj wrote: ============================================================ We have just acquired a used microscope made by ISI Inc. (their model alpha - 1980). Would anyone please have information on how we can obtain spare parts, filaments etc., for this machine. Many thanks for your help... ============================================================ The business of the ISI SEM's is now being handled by
Aspex Instruments LLC Formerly: RJ Lee Instruments Ltd. 175 Sheffield Drive Delmont, PA 15626 USA Tel: 1-724-468-5400 Fax: 1-724-468-0225 E-mail: pssales-at-rjleeinst.com
The former manager of the SEM operation when it was still ISI, Michael McCarthy, is now with Aspex. Michael might be the single most knowledgeable person in terms of spare parts for the column and vacuum system. Several of the main suppliers of consumables, like SPI Supplies, also offer filaments, new and retipped, apertures, and the other items of that nature you would be needing to maintain the microscope.
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: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I desperately want to be a materials researcher/Electron Microscopist. We have a Newly established Electron Microscope Laboratory. We need knowledge and skills. Please assist for a scholarship/suport as we have no funds, nor courses of the like at our University.
Hello everybody, We have an JEOL JXA 8600 EPMA in our Institute, with three WDS spectrometers. We are planning to buy an other one, and we where thinking about the H-type x-ray spectrometer.
Is there anybody in the list who has experience with this kind of spectrometers that could give me some information about them (their performance in general and also comparatively to standard spectrometers, etc.)?
Thanks
Laura Hernandez Laura Hernandez Laboratorio Microsonda Electronica Instituto GEA Universidad de Concepcion Casilla 160C Concepcion CHILE
Try using Vectabond treated slides. vectabond is available from vector laboratories. We had quite good results with leaf, stem, root sections sticking to these slides. Vector lab: 1-800-227-6666
Good luck,
Soumitra
I have no financial interest in Vector Laboratories.
On Sat, 31 Mar 2001, Rachel Spicer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello - } } Has anyone out there tried to adhere xylem sections to pre-coated } microscope slides like Fisher Probe-On Plus slides? I've tried and had a } zero percent section retention. I can imagine that this is because of the } scarcity of live cells (and plasma membranes). I've tried slow air drying } and various temperatures on the slide warmer. The sections are 15 microns, } and are from fixed (buffered paraformaldehyde) but not embedded samples. } I'm about to try Poly-L-lysine and amino-acyl silane treated slides, but } I'm not too hopeful because they (at least Poly-L) rely on the same } positively-charged surface principle. I want to avoid gelatin or albumin } subbing because I'm treating sections with protease, and also want to } minimize background staining. Any tips would be greatly appreciated. } } Rachel } } } } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } } }
You can do a brightness leveling image process with a number of a standard packages.
The procedure is simply to do a Gaussian blur that gives you the slowly varying component of the background and subtract that from your original image.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Feng Wu [mailto:fwu-at-bgumail.bgu.ac.il] } Sent: Wednesday, March 28, 2001 6:14 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: adjust the intensity of the image } Sensitivity: Confidential } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } Hi, All, } I take some diffraction contrast pctures. As the thickness of } the samples } changes sharply the brightness of the images is not uniform. } Does someone know } any software to adjust the brightness for the whole image? } Thanks inadvance. } Best regards. } Feng } ********************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } voice 972-8-6461473 } } fwu-at-bgumail.bgu.ac.il } ********************************************** } } }
I'm posting this message for a colleague; as always, I appreciate your help. Alice.
Alice Dohnalkova Environmental Microbiology Battelle, PNNL Richland, WA (509) 372-0692
} My graduate student has made TEM images of our plasma polymerized aniline } films. The films seem to have a "cauliflower" structure that could } probably be described as a fractal pattern. Have you ever made plasma } polymerized aniline films and if so did you see a cauliflower structure? } Your comments would be helpful. Thanks. } Pat } } Patrick D. Pedrow, pedrow-at-eecs.wsu.edu, www.eecs.wsu.edu/~pedrow
} Greetings, } I have found that polyethlyene-imine (PEI) is much stickier } than poly-lysine. I have no idea about the resin you mentioned, but } PEI is sticky stuff. It comes as a liquid. I make a 0.1% soloution in } ddwater, which I freeze in aliquots. Then I keep a working one in the } fridge. I coat coverslips in the stuff by floating them on a drop of } the PEI solution for about 10 sec, and then blotting off the excess } and letting them air dry. In that conditions, the coated 'slips are } good for at least months. } } Hope this helps, } Tobias Baskin }
Tobias:
Would this be Sigma catalogue number P-3143?
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
I am looking for a X-ray radiation detector used on TEMs. Currently we have a Geiger counter that is only sensitive to low energy X-ray. The purpose of the new detector is to sense high energy X-ray leakage from a 200 kV scope.
Any suggestion or clue will be greatly appreciated. Please contact off-line.
Dear Haifeng, The x-ray monitors on our HVEM are wrapped in a metalic sheath so they will be sensitive to the brehmsstrahlung spectrum of 1.2 MeV electrons. I don't think that they could be easily connected to your scope, but you might look into making a similar sheath for your existing Geiger counter. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
I have worked with CAMECA SX50 with the horizontal spectrometer. It was helpful when working with fractures, for example for identification of nonmetallic inclusions. Another it's advantage was that it was the only spectrometer with all 4 crystals, so it had more flexibility in maps or line scans acquisition. In quantitative analysis I did not use it both major and trace elements acquisition and never got any problems with its performance.
Vladimir Dusevich
-----Original Message----- } From: Laura Hernandez To: Microscopy-at-sparc5.microscopy.com Sent: 4/2/01 7:41 AM
Hello everybody, We have an JEOL JXA 8600 EPMA in our Institute, with three WDS spectrometers. We are planning to buy an other one, and we where thinking about the H-type x-ray spectrometer.
Is there anybody in the list who has experience with this kind of spectrometers that could give me some information about them (their performance in general and also comparatively to standard spectrometers, etc.)?
Thanks
Laura Hernandez Laura Hernandez Laboratorio Microsonda Electronica Instituto GEA Universidad de Concepcion Casilla 160C Concepcion CHILE
I am trying to determine a method to create or embed points of reference on an SEM sample. To give some background, the samples are sections of microprocessor packages that are placed in a modified stage with a three-point bend fixture. What I'm trying to do is monitor the movement of points on the specimen surface as the load is increased. I tried using the spot mode on the scope to see if I could remove some of the sputter coat, since most of the material underneath is non-conductive, but that was unsuccessful.
Imaging will most likely be done between 300 and 1000x. I need to have multiple reference points on the screen at one time, since I will be measuring relative displacements. The reference points do not need to be distributed in a uniform pattern.
I have tried applying some powder to the surface, since I read about this being done before, but the results were not acceptable. I may just be using the wrong type of powder, but I haven't been able to find out what kind of powders would work best
So, any ideas on how I may accomplish this?
Thanks,
Norman Kay Graduate Student AME Dept. The University of Arizona
I want to put a (cheap) video camera onto the optical microscope of my JEOL 840.
I want to leave the eyepiece lens on, so that users can have the option of easily removing the camera to use the microscope conventionally.
I have tried presenting several different models of CCD video cameras up to the eyepiece, both with and without the camera lens attached, but none gives me anything better than a smallish bright circle in the centre of the (black) field of view.
I presume that I need some sort of intermediate lens, but my understanding of physical optics has largely evaporated over the years.
Can someone point me towards a suitable text or other information source?
thanks
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
} From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } I want to put a (cheap) video camera onto the optical microscope of } my JEOL 840. } } I want to leave the eyepiece lens on, so that users can have the } option of easily removing the camera to use the microscope } conventionally. } } I have tried presenting several different models of CCD video cameras } up to the eyepiece, both with and without the camera lens attached, } but none gives me anything better than a smallish bright circle in } the centre of the (black) field of view. } } I presume that I need some sort of intermediate lens, but my } understanding of physical optics has largely evaporated over the } years. } } Can someone point me towards a suitable text or other information } source? } } thanks } } rtch
A CCD camera without a lens looking into and eyepiece usually has the opposite problem of having too much magnification. The image coverage of the image on the CCD camera can be increased when no lens is present on the camera simply by moving the camera further away from the eyepiece. My set up uses a 2.6x eyepiece for the video camera and a 10 x eyepiece to view the distance between the 2.6 eyepiece and the CCD element is about 2 inches or a little more and I have almost twice the magnification on the CCD camera as I see through the 10X eyepiece.
So just adding a spacer between your camera and your eyepiece should solve your problem.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
If you have coat your specimen with a thin carbon layer for conduction then add another layer of gold through a TEM grid as a mask you should be able to see the grid bars. Check out your local EM supplier's catalogue for the most suitable grid design.
Good luck, Ron
On Mon, 2 Apr 2001 15:48:48 -0700 SEM Machine {SEM-at-ACATC.AME.Arizona.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am trying to determine a method to create or embed points of reference on } an SEM sample. To give some background, the samples are sections of } microprocessor packages that are placed in a modified stage with a } three-point bend fixture. What I'm trying to do is monitor the movement of } points on the specimen surface as the load is increased. I tried using the } spot mode on the scope to see if I could remove some of the sputter coat, } since most of the material underneath is non-conductive, but that was } unsuccessful. } } Imaging will most likely be done between 300 and 1000x. I need to have } multiple reference points on the screen at one time, since I will be } measuring relative displacements. The reference points do not need to be } distributed in a uniform pattern. } } I have tried applying some powder to the surface, since I read about this } being done before, but the results were not acceptable. I may just be using } the wrong type of powder, but I haven't been able to find out what kind of } powders would work best } } So, any ideas on how I may accomplish this? } } } Thanks, } } Norman Kay } Graduate Student } AME Dept. } The University of Arizona } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
The magnifications you are using present the problem. I'm sure that you are also wanting as much resolution as possible from image processing of the resultant images.
Producing a non-conductive spot on the sample is a good idea, as it should stand out well, particularly during a slow record mode scan.
My vote - copier or laser printer toner. Very small particle size, non-conductive plastic composition. Also, once the powder is sprinkled on, it can be adhered to the surface with a little heat to ensure that it doesn't move around.
On Monday, April 02, 2001 5:49 PM, SEM Machine [SMTP:SEM-at-ACATC.AME.Arizona.edu] wrote: } } } I am trying to determine a method to create or embed points of reference on } an SEM sample. To give some background, the samples are sections of } microprocessor packages that are placed in a modified stage with a } three-point bend fixture. What I'm trying to do is monitor the movement of } points on the specimen surface as the load is increased. I tried using the } spot mode on the scope to see if I could remove some of the sputter coat, } since most of the material underneath is non-conductive, but that was } unsuccessful. } } Imaging will most likely be done between 300 and 1000x. I need to have } multiple reference points on the screen at one time, since I will be } measuring relative displacements. The reference points do not need to be } distributed in a uniform pattern. } } I have tried applying some powder to the surface, since I read about this } being done before, but the results were not acceptable. I may just be using } the wrong type of powder, but I haven't been able to find out what kind of } powders would work best } } So, any ideas on how I may accomplish this? } } } Thanks, } } Norman Kay } Graduate Student } AME Dept. } The University of Arizona } } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
My collegue C. Ulhaq want to know who practice Lorentz Microscopy on TEM (in Europe particulary).
The questions would be : which kind of microscope you use, do you use special polar pieces, and did you buy it or were they "home made". Same question about the sample holder. What are the max magnification accessible ? We have a ABT Topcon 002B, with the the possibility to change the polar pieces.
You can answer direct to my collegue (corinne.ulhaq-at-ipcms.u-strasbg.fr), or on the list.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
Thanks for the replies and the advice that you have offered.
I have Some additional information. I am a BSc.[Physics, Mathematics] holder. Currently I am working at the E.M laboratory of the university of Dar es Salaam as a supporting staff, and I am looking for the opportunity to pursue an MSc. and hence a Ph.D that will qualify me to research fellow. Funds are a hindrance. We have one old ZEISS9S2 TEM, and a new LEO 910 analytical TEM. An MSc. course in TEM or a research in metals/ceramics will do.
Hello, We have a client who needs to embed a cell culture in paraffin. In our lab we embed cultures in agarose and then embed for TEM. We do not have experience embedding paraffin. My question is, since xylene is used for paraffin how do you keep the culture from dispersing. Any suggestions? Thank you
Karen Kelley Senior Electron Microscopist University of Florida ICBR Electron Microscopy Core Lab Box 118525 Gainesville Florida Lab: 352-392-1184 fax: 352-846-0251 email: klk-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/staff/karenpage.html
I have not done Lorentz microscopy (here LEM) in a long time and I am not very familiar with the configuration of the Topcon 002B. This is what I remember ,for LEM to work the sample has to be outside the strong magnetic field of the OL pole piece. In the good old days, we used a modified top entry-type specimen holder which was longer than usual so that the specimen would sit below its normal position. In addition, the IL electronics had to be modified so that focus could still be attained when the specimen was in this lower position. We normally used magnifications of up to about 20 KX if I recall.
The second way of doing this is to essentially turn off the objective lens , but you have to be able to focus with the IL . Some of the newer scopes might not be able to do this without modifications to the electronics. In our case all the modifications were done by the manufacturer.
Hope this helps
Jordi Marti
-----Original Message----- } From: Faerber Jacques [mailto:Jacques.Faerber-at-ipcms.u-strasbg.fr] Sent: Tuesday, April 03, 2001 3:25 AM To: Microscopy Society of America
Hi everyone, We are working with suspension cells (Jurkats or H9s) and we need a protocol to prepare them for fluorescent microscope (both fixed and live cell imaging). Thanks in advance for your help.
Asli Oztan
asost2-at-pitt.edu University of Pittsburgh Molecular Virology and Microbiology
You can try spinning (centrifuging) the cells down so they become a packed ball of cells, then carefully put them into a bag (i think they are nylon bags) for processing and subsequently into paraffin block.
Frank Lee
Karen Kelley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello, } We have a client who needs to embed a cell culture in paraffin. In our lab } we embed cultures in agarose and then embed for TEM. We do not have } experience embedding paraffin. My question is, since xylene is used for } paraffin how do you keep the culture from dispersing. Any suggestions? } Thank you } } Karen Kelley } Senior Electron Microscopist } University of Florida } ICBR Electron Microscopy Core Lab } Box 118525 Gainesville Florida } Lab: 352-392-1184 fax: 352-846-0251 } email: klk-at-biotech.ufl.edu } http://www.biotech.ufl.edu/~emcl/staff/karenpage.html
Dear Norman, When we had a similar problem of creating a reference for studying crack growth, we used the gold-evaporated grid method, as Ron Doole mentioned. However, this gave problems because the grid is uniform, so after you've traveled a little way, you had no unique reference to keep you placed. We solved this by also puting a drop of latex sphere suspension on the surface, before sputter coating. This is a suspension of latex spheres of specific size, I believe we used one micron, but you can use a size suitable to your magnification. The suspension dries to form a random pattern of dots that can be compared in photos. You amy have to experiment with the concentration of spheres to get the right coverage. We were lucky enough to have a researcher making latex spheres who gave us some of her duds, but these suspensions can be purchased. I hope this helps. At 03:48 PM 4/2/01 -0700, you wrote: } } I am trying to determine a method to create or embed points of reference on } an SEM sample. To give some background, the samples are sections of } microprocessor packages that are placed in a modified stage with a } three-point bend fixture. What I'm trying to do is monitor the movement of } points on the specimen surface as the load is increased. I tried using the } spot mode on the scope to see if I could remove some of the sputter coat, } since most of the material underneath is non-conductive, but that was } unsuccessful. } } Imaging will most likely be done between 300 and 1000x. I need to have } multiple reference points on the screen at one time, since I will be } measuring relative displacements. The reference points do not need to be } distributed in a uniform pattern. } } I have tried applying some powder to the surface, since I read about this } being done before, but the results were not acceptable. I may just be using } the wrong type of powder, but I haven't been able to find out what kind of } powders would work best } } So, any ideas on how I may accomplish this? } } } Thanks, } } Norman Kay } Graduate Student } AME Dept. } The University of Arizona } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Can someone identify a supplier for small amounts of methlyamine tungstate powder (used for negative staining in TEM)? I have tried EMS, Ted Pella, and Polysciences (the supplier of our original decade-old vial). Robert J. Palmer Jr., Ph.D. Natl Inst Dental Craniofacial Res - Natl Insts Health Oral Infection and Immunity Branch Bldg 30, Room 308 30 Convent Drive Bethesda MD 20892 ph 301-594-0025 fax 301-402-0396
I'm trying to get a handle on the value of an old Auger/ESCA spectrometer. The model is PHI 558, by Perkin-Elmer. It was built around 1985 or 86, and the electronics are quite old, but it's intact and fully functional. It also has a LEED detector and has been upgraded over time. A partial list of parts is included below. Does anyone have an idea what this might be worth?
Thank you, Ellen Carrillo-Heian
emheian-at-engr.ucdavis.edu Dept. of Chem. Eng. and Mat. Sci. UC Davis Davis, CA 95616 USA ---------------------- Partial list of components:
} 32-010 Lock In Amp } 20-0275 Electron Multiplier Supply } 32-095 X Ray Source Control } 22-040 DC Power Supply } 16-020 Heat Exchange / Deionizer } 20-805 Analyzer Control } 32-100 Electron Multiplier Supply } 11-065 Ion Gun Control } 20-115 Ion Gun Control } 11-010 Electron Gun Control } 11-055 ESCA / Auger System Control } 11-500A Auger System Control } Inficon 012-214 } 04-303 Differential Ion Gun } 04-548 Dual Anode X-ray Source } 15-255 GAR Precision Electron Energy Analyzer } Ultek DI Pump } 218075B-26 UHV Instruments (Tag in front of the screw/bellows assembly, on } frame.) } } And a 386 Compaq controlling it. } } The chamber has the dual chamber translation set up. (With the 4 ft } screw/bellows) }
We do this using contamination spots from the microscope itself. We put down a grid array of spots using our Isis system to control the stage and spot positons. Focus the beam to a spot and let is sit there for a minute or so and a contamination spot should appear. We are using electropolished aluminum. I dont know if it will work for your material but its worth a try Then we strain our material and look at it again. Works well.
I would like to do high resolution TEM on spherical, semiconductor nanoparticles. The diameters of these particles range from 1 to 10 nm. Our goal is to acquire good digital images of these particles so that we can size them in house. Please let me know if you can help and how much you charge per sample or per hour. Your help will be greatly appreciated.
Dear Dr. Palmer, Nanoprobes sells the methylamine tungstate you desired under the name "NanoW". It is an excellent negative stain. They also make methylamine vanadate, a similar, but lower atomic number stain they call "NanoVan". More information is at www.nanoprobes.com. J. Hainfeld
Dr. James F. Hainfeld Brookhaven National Laboratory Biology Dept. Bldg. 463 Upton, NY 11973 USA Tel. 631-344-3372 Fax. 631-344-3407 email: hainfeld-at-bnl.gov website: http://bnlstb.bio.bnl.gov/biodocs/stem/stem.html
Generally, equipment depreciates at the rate of 30% per year on the balance. In other words, an SEM with an initial cost of 100K is worth 70K after the first year, 49K the second year, 34.3k the third year, etc.
These numbers are based upon the experience I have had with used equipment sales during the past five years. Other allowances are made for equipment that has been abused, etc. Strangely enough, accessories add little to the resale value of equipment.
I am sure there are others who would differ as it would not fit into their accounting procedures but my experience has been that scientific equipment depreciates more than computers.
Regards,
Earl Weltmer
I have no financial interest in this thread only experience.
----- Original Message ----- } From: "Ellen Carrillo-Heian" {emheian-at-engr.ucdavis.edu} To: "microscopy" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, April 03, 2001 11:25 AM
Hi everybody (again),
I am looking for a program that allows me to quantify a given group of nanoparticles that have various shapes and sizes. These particles range from 1 to 10 nm in diameter. Your help will be greatly appreciated. Thank you.
We sell this as a 2 % aqueous solution (suitable for use directly) - the product is called "Nano-W." It's listed on our web site catalog under "Negative staining."
Regards,
Rick Powell
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
What kind of computer depreciates (looses value) slower than scientific equipment? A regular PC or Mac drops by over 50% the first year. After an additional six months, the system worth next to nothing. But of course, the new and improved model is $2K or more. Either way, the standard IRS depreciation schedule for computers and scientific equipment is five years. I would say that this is more appropriate for scientific equipment than it is for computers. But YMMV.
gary g.
At 05:16 PM 4/3/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This is an interesting thread. Our unit is supposed to operate at "full cost recovery", and as such we are charged depreciation of the instruments (funded from a central equipment grant), which are depreciated over 15 years. Only computers are depreciated over 3 years. If anyone else is aware of some vaguely standard depreciation times for TEMs, SEMs, confocal, light microscopes, I'd appreciate hearing about this.
Thanks, rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
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Rosemary said - } This is an interesting thread. Our unit is supposed to operate at "full } cost recovery", and as such we are charged depreciation of the instruments } (funded from a central equipment grant), which are depreciated over 15 } years.
FWIW, I understand that that our organization (A Canadian federal government one) also depreciates this kind of major capital equipment over 15 years. (Except, apparently, helicopters - our Navy is still using ones which are, literally, older than most of the pilots flying them....)
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
John Russ recently demonstrated an adaptive equalization technique for us that works quite well. Basically it is histogram equalization over a local area of the image that is then stepped over the whole image. He has implemented this method in his IP Toolkit and Fovea Pro packages that plugin to PhotoShop. It is also described in The Image Processing Handbook, 2nd edition on page 222.
Just a satisfied customer...
Henk Colijn
At 11:42 AM 4/2/01 -0400, Walck, Scott D. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Fools are pleased when they discover error. The wise are pleased when they discover truth.
An interesting thread indeed! We are in the midst of surplusing out a lot of used histology equipment, and find that there are two different methods of determining value. Depreciation of equipment for tax purposes is done according to state/federal tax laws--and I think that means after 5 or 10 years (the time seems to change, and I can never keep up with) the _tax_ value is zero. However. When it comes to disposing of the equipment, our business folks are using a different "book value" that has values significantly greater than current market value. So.... Draw your own conclusions.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On Wed, 4 Apr 2001 15:47:39 +1000, Rosemary White wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | This is an interesting thread. Our unit is supposed to operate at "full | cost recovery", and as such we are charged depreciation of the instruments | (funded from a central equipment grant), which are depreciated over 15 | years. Only computers are depreciated over 3 years. If anyone else is | aware of some vaguely standard depreciation times for TEMs, SEMs, confocal, | light microscopes, I'd appreciate hearing about this. | | Thanks, | rosemary | | | Rosemary White | Microscopy Centre | CSIRO Plant Industry | GPO Box 1600 | Canberra, ACT 2601 | Australia | | phone 61-2-6246 5475 | fax 61-2-6246 5000 | email r.white-at-pi.csiro.au | | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
I think that CSIRO is on the high side. In the Canadian government 10 years seems to have been the semi-official standard for as long as I can remember. In my lab, though, we informally think of two 'effective lifetimes', 7 years or thereabouts for the TEM and SEM, and 10 for the other beam instruments (EPMA, SIMS, XPS and SAM). Furthermore, we regard these as upper limits for two reasons:
- vendors these days are operating on ever-tighter parts inventories, thus a beam instrument may have some good years left in it, but you find you can't get a crucial part any longer. This happened to us recently with our 12 year old Cameca SIMS, when a flight tube developed ultrafine cracks and we discovered that they are not kept in stock any more. After a lot of arm-twisting and calling of favors, we tracked down one of the few remaining ones in Europe, otherwise we would have been down for 3 months awaiting a custom-built one.
- we do a lot of work with the private sector, some on a contract basis. They often come to us to get state-of-the-art data quality which has been collected in a timely fashion. The first is what usually gets their attention in the first place, while the second is crucial to keeping their attention.
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 613-992-8735 email: malis-at-nrcan.gc.ca
} ---------- } From: Rosemary White } Sent: Wednesday, April 04, 2001 1:47 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: equipment depreciation } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } This is an interesting thread. Our unit is supposed to operate at "full } cost recovery", and as such we are charged depreciation of the instruments } (funded from a central equipment grant), which are depreciated over 15 } years. Only computers are depreciated over 3 years. If anyone else is } aware of some vaguely standard depreciation times for TEMs, SEMs, } confocal, } light microscopes, I'd appreciate hearing about this. } } Thanks, } rosemary } } } Rosemary White } Microscopy Centre } CSIRO Plant Industry } GPO Box 1600 } Canberra, ACT 2601 } Australia } } phone 61-2-6246 5475 } fax 61-2-6246 5000 } email r.white-at-pi.csiro.au } } }
I am processing samples of white fat for TEM and have been having trouble with a dense precipitate that covers the cytoplasm. Nuclei and blood vessels are relatively unaffected. So far I've tried 3 different fixatives: Trump's and 2.5% glut/2% para in either 0.1M cacodylate of 0.1M phosphate buffer. The samples have been osmicated for one hour and embedded in either Spurr's resin or Epon. The precipitate is present in unstained sections as well as those stained with UA alone or UA and lead. Treatment with .5N HCl for 2 minutes alleviates the problem somewhat but bleaches the sections so much that they are very difficult to see. If anyone has encountered similar problems and has suggestions for me I would be most grateful.
Thanks in advance
Germaine G. Boucher TEM Lab Pfizer Global Research and Development Groton, CT
We are looking to replace a very old EDS system on our Hitachi S-2460N variable pressure SEM. I would like to know about your preferences for and experiences with different companies, both good and bad. Please reply directly to me. If anyone else is interested, I could submit a summary to the list after I've collected all the responses.
Thanks in advance.
Jean Ross Central Microscopy Research Facility University of Iowa
I have two questions concerning service contracts on confocals. Let me start by saying i have had a confocal for about 8 years and would never consider going without one. I have a Biorad 2000 going off warranty and need to make a decision.
First question: Biorad no longer guarantees a response time - they now promise to get to you as fast as they can but no longer promise a 48 or 72 hr response. Have other confocal manufacturers done this also?
Second question: My university is pushing replacing service contracts with "insurance" contracts with a major vendor who then pays for a service visit from the manufacturer on an hourly basis. All parts, travel, service repair time, etc are covered at a price that is typically 75% less than the manufacturer's service contract. They guarantee the price and coverage for 3 years. Personally I don't know how they could make money on this deal since we average a fair number of visits and spare parts (e.g. lasers) in a typical year. Does anyone have experience with this type of situation with confocals? The company the University is dealing with is CIC but there are several other ones out there.
Thanks for any input. -- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
dear listservers.... i need some advice.... we use Permount as our mountant...however, as we cut our slides extremely thick (120 micra) for our staining method (Golgi-impregnations of neurons), it often takes 2-3 weeks for the Permount to dry sufficiently so that we can actually use the slides without it getting on our microscope stage, or the coverslip moving around under our oil-immersion lenses.... so....my question is --- does any microscope maven out there know if there is anything that can be done to accelerate the drying/hardening the Permount....??? many thanks for any help.... regards, Ron Mervis ~~~~~~~~~~ Ronald F. Mervis, Ph.D. Neuro-Cognitive Research Laboratories 2109 West Fifth Avenue Columbus, Ohio 43212 USA ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ..independent nonprofit contract laboratories dedicated to quantitiative neurostructural analysis to promote our knowledge and understanding of human neurological diseases, neurodegeneration, and neuroplasticity.... ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Tel: (614)-486-6404; lab: (614)-486-6080 Fax: (614)-486-6020 e-mail: RonMervis-at-aol.com (or) RonMervis-at-Neuro-Cognitive.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ "...can the human soul be glimpsed through a microscope? Maybe, but you'd definitely need one of those very good ones with two eyepieces." - Woody Allen
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} dear listservers.... {BR} i need some advice.... {BR} we use Permount as our mountant...however, as we cut our slides extremely {BR} thick (120 micra) for our staining method (Golgi-impregnations of neurons), {BR} it often takes 2-3 weeks for the Permount to dry sufficiently so that we can {BR} actually use the slides without it getting on our microscope stage, or the {BR} coverslip moving around under our oil-immersion lenses.... {BR} so....my question is --- does any microscope maven out there know if there is {BR} anything that can be done to accelerate the drying/hardening the {BR} Permount....??? {BR} many thanks for any help.... {BR} regards, {BR} Ron Mervis {BR} ~~~~~~~~~~ {BR} Ronald F. Mervis, Ph.D. {BR} Neuro-Cognitive Research Laboratories {BR} 2109 West Fifth Avenue {BR} Columbus, Ohio 43212 USA {BR} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ {BR} ...independent nonprofit contract laboratories dedicated to quantitiative {BR} neurostructural analysis to promote our knowledge and understanding of human {BR} neurological diseases, neurodegeneration, and neuroplasticity.... {BR} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ {BR} Tel: (614)-486-6404; lab: (614)-486-6080 {BR} Fax: (614)-486-6020 {BR} e-mail: RonMervis-at-aol.com (or) RonMervis-at-Neuro-Cognitive.org {BR} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ {BR} "...can the human soul be glimpsed through a microscope? Maybe, but you'd {BR} definitely need one of those very good ones with two eyepieces." {BR} - Woody Allen {BR} {/FONT} {/HTML}
I have seen some beautiful SEM pictures of semiconductor interconnect lines (Copper and aluminum) in which the dielectric material (Silicon dioxide I assume) has been completely removed. Can someone tell me what was used to remove the dielectric without affecting the metallization?
Hello friends, I have a gold bronze composition material coming in for imaging by SEM, x-ray mapping, and optical microscopy. I could us a sample prep recommendation, particularly for a etchant or so I think. This is primarily a failure analysis project in which we want to clearly observe and differentiate grain boundaries. Recommendations would be greatly appreciated.
thanks, Bruce Brinson Optical Analyst Rice University
Germaine, If you don't rinse in buffer well enough after the initial fixation , the glut will form a precipitate with osmium. Try washing 4x or 5x for 15 minutes each between glutaraldehyde and osmium. Good luck,
At 10:34 AM 4/4/01 -0400, Boucher, Germaine G wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
Thanks for all the recommendations for suppliers of cameras and interfaces, but what I was wanting was a pointer to a text or somesuch from which I could figure out myself what I need.
There must be someone in this learned and experienced community who's been there and done that, isn't there?
"That", for those who may have missed my first post, being the problem of how to work out what sort of intermediate lens would be needed to interface a small cheap CCD video camera (or a webcam) so that it gives a good image when looking into the existing eyepiece lens of a given optical microscope (in this case the OM of a JEOL 840 SEM).
thanks
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Dear colleagues, I would like to buy a digital camera for my Axiolab Zeiss microscope. Unfortunately I am a bit confused in the amount of available data. I would need a digital camera of the resolution that matches the quality of film cameras in order I need not scan photos or negatives. Could you please be so kind and give me a piece of advice? I have Olympus Camedia 3030 with 3,34 mil pixels. Will this camera and resolution do or do I need to buy another one? If yes of what resolution and type? I will be very indebted for an advice because I receive often contradicting information for different distributors and I am not much wise about it. Many thanks in advance. With best wishes Jiri Kalvoda Department of Geology and Paleontology Masaryk University Kotlarska 2 61137 Brno Czech republic
It is easily done with a plasma etch using CF4 + O2.
The top layer of "glass" is typically silicon nitride over silicon dioxide or in earlier devices, it is boron phosphor silicon glass (BPSG).
I have some colorized shots on my web site at:
http://photoweb.net
More get added and some get swapped over time.
gary g.
At 11:48 AM 4/4/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Rod McCabe wrote: ======================================================= I have seen some beautiful SEM pictures of semiconductor interconnect lines (Copper and aluminum) in which the dielectric material (Silicon dioxide I assume) has been completely removed. Can someone tell me what was used to remove the dielectric without affecting the metallization? ======================================================= While HF and Q-Tips can be used to swab off the SiO2, the better way (in our opinion) is with reactive plasma etching, using CF4 as the reactive gas. This way the layer is removed in a way that does not disrurb, for example, corrosion product that might have formed underneath. If you use the wet chemical approach, you can dissolve and swab away features of interest, such as corrosion product. And you have removed that which you might otherwise have been able to analyze with EDS or even Auger.
Several manufacturers offer table top plasma etchers, SPI Supplies being one of them. You can find information about the SPI Plasma Prep™ II on our website given below.
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: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I am writing for comments -- off-list please -- about the CleanZone LF clean air workstation manufactured by IQAir, which, I am told, is used in Switzerland and Germany, but only recently has been introduced in the United States. With thanks,
James Martin Orion Analytical, LLC www.orionanalytical.com martin-at-orionanalytical.com
A colleague has asked for recommendations for setting up a digital darkroom (fun to spend someone else's money!). This person would benefit from a really good scanner that could deal with prints, large format negatives (4"x5", 3.25"x4") as well as 35 mm slides. At one time I looked into an Agfa Duoscan T2500. Do any of you have an opinion about this or other suitable scanners?
I know this subject comes up regularly, but I don't feel bad about introducing it again, since technology evolves so quickly!
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
thanks to everyone for the advices about all type of glues for tripod polishing. I tried to make some sort of summary from the different answers I have received. I would be happy if it could be any use for other tripod beginners as well.
Cheers! Csaba
1.) The best mail which decribes well the everyday life and joy of a microscopist comes first.
You really can't look at a spec sheet and know that it works - it's all trial and error.
2.) About the type of glue to be used:
The best material to use is super glue which is a cyanoacrylate material. There are different types available under various names. They all have slightly different properties and vary in such things as viscosity. The most important point is that the super glue is fresh. Once it is opened, you can use it for a short time, but then need to replace it. Sometimes, the glue from an unopened package can also be bad if it has been on the shelf for a long time. The trouble with using consumer products is that they sometime change without warning.
You should buy it from a place that has a high turnover rate of it so that it is fresh. The IBM guys recommend keeping the stuff unopened in the refrigerator for any length of time. Once it has been opened, you can't use it for very long (a day or two). I have used the Loctite a little longer because it has a very good sealing top. The Loctite product is also available in a "pen" type applicator which seems to have the best bench life of any of the applicators I've used. My tube usually is swiped off the bench long before it goes bad.One of the advantages to the cyanoacrylic cement is it dissolves, in a reasonable time, in acetone.If you were to use a crosslinked epoxy, you'll need to devise a cleaning scheme that will exhaustively remove the epoxy without altering your sample.
Basically what we're looking for is an intermediate viscosity glue (somewhere between maple syrup and water) that bonds in about 30-40 seconds and is very strong. Don't use any type of super glue gel! It doesn't work, you have to use the thin stuff.
In the end we are using a Loctite Prism 460.
Crystalbond 509. This is an acetone soluble glue which melts at 150*C and becomes quite viscous. We use it quite often here at Queens as a temporary glue for ceramics that have to be ground and polished on all four sides. If the bond breaks you simply heat it up and reset the part. This stuff comes in sticks that last for a very long time. Their website is as follows. {http://www.aremco.com/}
I like to glue specimens with epoxy resin because the substance does not harden too fast, and you always have time to find the best location for your sample, so that the latter does not break.
I have tried the "super glue" approach with no luck. Many technicians advocate Lock Tite brand of super glue that many companies sell with the tripod polish kit. I have always utilized Crystalbond adhesive. It is a low melting point (77 C) wax that is dissolved in acetone. If your sample is heat sensitive, super glue is the only other choice I know.
3.)Glueing advices:
--the biggest reason for sections falling off is the cleanliness of the pedastal. It must be cleaned with clean solvents that do not leave any trace of a contamination film on the glass. Check for cleanliness by holding the tool such that light reflects from the surface.
--the suface that you are glueing to should be as rough as possible to obtain a tooth or larger surface area for the glue to adhere to.
--any way you can improve the surface area would be great.
--the same above would be for the specimen
--pressure on the sample while it cures might be important, but the IBM guys just wick the extra stuff away from the sample and don't put a lot of pressure on the sample.
-- ____________________________________________ Csaba Cserhati Univ.of Debrecen / Dept. of Solid State Phys. Hungary tel/fax: 36 52 316073 e-mail: cserhati-at-delfin.klte.hu ____________________________________________
I've followed with interest the discussion on equipment depreciation and service contracts . I've noticed over the years in both metals research and now the semiconductor industry with increasing sophistication/specialisation of equipment running a lab to a given budget seems to mean accepting manufacturers service contracts with fewer 'independent' sources being able or willing to offer maintenance assistance .
My question arises from a recent 'confocal service contract' letter and I wonder does an insurance contract service alternative as offered by CIC exist in the UK ? and if so could anyone let me know where I could get more information ? .
Martyn Harris Device Engineer ESM Ltd , Cardiff Rd Newport , South Wales UK NP10 8YJ .
First, a disclaimer. I am a third party service provider for a variety of equipment, but not confocals. I have worked for a number of manufacturers in the past and been self-employed for around 20 years.
In regards to the first question - service departments in general are getting squirrellier. Contract prices have gone up greatly over the last decade or so while quality and parts stores have gone down. A generalization, granted, but one I'd be happy to back up.
As for the second question - don't get me started again. Given recent problems with the industry in general, I can only suggest that your university carefully study the question and check with several of their customers who have been with them for at least a couple of years insuring similar equipment. That goes for any such provider. I've been a vocal antagonist here of the concept, and from what I have heard from some customers, perhaps rightfully so. Not right, so far, in my feelings regarding the potential long term problems. Rather in the broad approach they have taken. Perhaps in trying to be a jack of all trades, they are a master of none.
To those of you who might be surprised by my abnormally low tone in this posting, please understand that we are getting to a point in time where these organizations are getting quite large in a very short period of time. As with any company or industry, they do have problems that they will not publicize. They also have increasingly large budgets for legal teams that would probably be anxious to root out any libel. They do save many organizations large amounts of money, but you have to ask, at what cost?
On Wednesday, April 04, 2001 10:20 AM, Tom Phillips [SMTP:PhillipsT-at-missouri.edu] wrote: } } } I have two questions concerning service contracts on confocals. Let } me start by saying i have had a confocal for about 8 years and would } never consider going without one. I have a Biorad 2000 going off } warranty and need to make a decision. } } First question: Biorad no longer guarantees a response time - they } now promise to get to you as fast as they can but no longer promise a } 48 or 72 hr response. Have other confocal manufacturers done this } also? } } Second question: My university is pushing replacing service } contracts with "insurance" contracts with a major vendor who then } pays for a service visit from the manufacturer on an hourly basis. } All parts, travel, service repair time, etc are covered at a price } that is typically 75% less than the manufacturer's service contract. } They guarantee the price and coverage for 3 years. Personally I } don't know how they could make money on this deal since we average a } fair number of visits and spare parts (e.g. lasers) in a typical } year. Does anyone have experience with this type of situation with } confocals? The company the University is dealing with is CIC but } there are several other ones out there. } } Thanks for any input. } -- } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax) } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
I don't know if this is exactly what you are looking for, but Reetz and Coworkers have recently published a high troughput routine for nanaoparticle analysis: Reetz, Manfred T.; Maase, Matthias; Schilling, Tobias; Tesche, Bernd. Computer Image Processing of Transmission Electron Micrograph Pictures as a Fast and Reliable Tool To Analyze the Size of Nanoparticles. J. Phys. Chem. B (2000), 104(37), 8779-8781.
Good Luck,
Andreas
************************************************* Dr. Andreas Taubert Dept. of Materials Science and Engineering 3231 Walnut Street University of Pennsylvania Philadelphia PA 19104-6272 tel: +1 215 898 2700 fax: +1 215 573 2128
Physical Chemistry is everything for which 1/T is linear ... *************************************************
Tina: We have the Duoscan 1200 but the 2500 is also a very nice unit--additional (real) resolution and a high O.D. range plus 14 or 16 bit image depth. The Agfa units also come with a built-in transparency plate (rather than having to add on a separate transparency adaptor). I find that color fidelity is very good with the Duoscan, and Agfa provides both reflective and transparency calibration standards. I am currently scanning in 3x4 TEM negatives at 12 bits (yield is about 26MB per image), and scan time is fairly rapid. There is one option that you might want to consider: the DIImage unit is made for up to 4x5 negatives, and I think (can't remember the last time I read the specs--the neurons aren't firing today) that resolution is in the 2700 dpi range--even for the 4x5 size.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
The opinions expressed are solely my own and do not constitute an endorsement of any vendor or manufacturer. I have no fiduciary interest in either company.
On Wed, 4 Apr 2001 17:22:23 -1000 (HST), Tina Carvalho wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Hi, All- | | A colleague has asked for recommendations for setting up a digital | darkroom (fun to spend someone else's money!). This person would benefit | from a really good scanner that could deal with prints, large format | negatives (4"x5", 3.25"x4") as well as 35 mm slides. At one time I looked | into an Agfa Duoscan T2500. Do any of you have an opinion about this or | other suitable scanners? | | I know this subject comes up regularly, but I don't feel bad about | introducing it again, since technology evolves so quickly! | | Mahalo, | Tina | | http://www.pbrc.hawaii.edu/bemf/microangela | **************************************************************************** | * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * | * Biological Electron Microscope Facility * (808) 956-6251 * | * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* | **************************************************************************** | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
We have a professor here who has a 1 cm x 2.5cm steel layer about 100 um in thickness on a ceramic substrate. The metal layer was sputter deposited onto the ceramic substrate. The ceramic substrate extends past the metal layer. He needs to get a thickness gradient across the steel layer along the 2.5 cm length. He would like to have one end at about 50 microns in thickness and the other at 2 microns with a gradually decreasing thickness gradient. Steps down would be ok although a smooth transition would be better. We have a laser profilometer to measure anything that we produce.
Does anyone out there have any ideas how to do this? Would a tripod polisher work? I thought about electropolishing and masking off portions at a time but I worry about what will happen at the interface between the ceramic and the metal. Could we alter a dimpler?
Nominal grain size of film is about 10 microns (varies with film speed etc but this is the right order of magnitude). Thus to digitize the film to it's nominal limits your scanner should be able to digitize to better than this spatial dimension.
A simple back of the envelope calculation says a spatial resolution of 10 microns is 2540 - dpi..... and as we all know that must be the optical resolution of the scanner not the interpolated resolution. Scanners at this end are obviously more than you need to digitize photo's and get expensive quickly. Also when you see 2 numbers listed as the scanners resolution, believe only the first number, that is the CCD resolution.
Now add your bit depth. 12 bits is the minimium I would shoot for greyscale image, but if your attempting diffraction work the higher the better (i.e. 14 -16 bits+). For color work obviously multiple the bit depth by 3 one for each primary color (RGB). I've seen a number of 36 bit color scanners but not too many 48 bit ones at } 2540 dpi.
Lastly, dit depth is irrelevant if you don't have a high optical density capabilities otherwise your just digitizing noise. The highest value I believe is an OD of 4.0 but this is for DRUM scanners. Flatbed scanners typically run as low as 2.8, upwards to about 3.4 for the best I've seen in a flatbed.
At 09:40 AM 4/5/01 +0000, Ritchie Sims wrote: } "That", for those who may have missed my first post, being the } problem of how to work out what sort of intermediate lens would be } needed to interface a small cheap CCD video camera (or a webcam) so } that it gives a good image when looking into the existing eyepiece } lens of a given optical microscope (in this case the OM of a JEOL 840 } SEM).
You might try a web search for "afocal coupling". See http://www.photosolve.com/xtendascope.asp for example.
This page, and the message below, are for coupling to the eyepiece of telescopes, but the process should be similar with a microscope eyepiece. If you're looking for "small and cheap", put in a wide field eyepiece and hold the camera close to it and see what happens. I suspect you'll have some cropping of the image due to optical mismatch.
- John
} A friend of mind recently bought a fairly expensive SLR digital still } camera, an Olympus C-2500L, } } http://www.olympusamerica.com/product.asp?c=57&p=16&s=12&product=380 } } and has experimented with afocal photography through both his 8-inch f/6 } Dob and a microscope. All he sees in the camera is a small central disk } of light. He got the following explanation from a post on rec.photo. } digital. } } For cameras with LARGE taking lenses (f 2.8, f2.0) [...] there will } be a problem since their entry pupil ( the area of the lens that lets } the image into the lens) is so large compared to the exit pupil (the } opening on the eyepiece that lets the image out of the microscope) } that a large amount of the image will be lost!! This results in } severe vignetting, and so only a small central spot of image in a } dark field is recorded by the digital camera. } } Does this sound reasonable? The optics of this situation are pretty } mysterious to me, so I can't judge. But it seems like increasing the } focal length of the camera lens (zooming in) should compensate for the } effect of the small exit pupil, and that the problem might be more one } of eye relief (he just can't get the lens close enough to the eyepieces, } which I think are a couple of Plossls and the 25mm SMA that comes with } Celestron Dobs).
And here's the response so far.
} Subject: Re: afocal astrophoto problem: exit pupil? } Date: Wed, 4 Oct 2000 17:52:19 -0400 } From: "Michael A. Covington" {See http://www.CovingtonInnovations.com for address} } Organization: MindSpring Enterprises } Newsgroups: sci.astro.amateur } } } Does this sound reasonable? } } No. The camera lens is *supposed* to have a larger entrance pupil than the } exit pupil of the telescope. When doing afocal photography with an SLR the } entrance pupil of the lens is an inch or more in diameter. } } If there is vignetting, it's probably because the camera is the wrong } distance from the eyepiece -- either too close or too far. } } -- } } Clear skies, } } Michael A. Covington / AI Center / The University of Georgia } Author, ASTROPHOTOGRAPHY FOR THE AMATEUR } http://www.CovingtonInnovations.com/astro {} { } } } Subject: Re: afocal astrophoto problem: exit pupil? } Date: Wed, 4 Oct 2000 14:25:48 -0700 } From: "Bob May" {bobmay-at-nethere.com} } Organization: Posted via Supernews, http://www.supernews.com } Newsgroups: sci.astro.amateur } } Bet that when you look at the viewfinder, you will see an image just } like one that you would see if your eye were at a certain distance } from the EP. If that image looks like the eye is too far away from } the EP then it's a sure thing that the camera's lens is too far away } from the EP. That's how it's all done. The camera is nothing more } than an aritificial eye. } -- } Bob May } Remember that computers do exactly what you tell them to do, not what } you think that you told them! } Bob May } } } Subject: Re: afocal astrophoto problem: exit pupil? } Date: Wed, 04 Oct 2000 19:43:43 GMT } From: "Chuck Olson" {chuckolson01-at-home.com} } Organization: -at-Home Network } Newsgroups: sci.astro.amateur } } Yes, it is critical that the eyepiece and camera lens be somewhat } physically compatible with each other. The eyepiece must put the } exit pupil about in the plane of the camera iris opening, which } in most instances requires the eyepiece to be virtually in } contact with the camera lens front element. For instance, the } Nikon Coolpix 950 and 990 have relatively small lens fronts and a } nice 28mm (I think) thread that adapts readily to the T-thread } that is often used in astrophotography. As a result, the CP950 } easily looks through 17mm , 26mm, or 32mm Plossl eyepieces. Even } there, as you point out, the camera needs to be operating at the } tele end of its zoom range to fill the rectangular frame, rather } than showing a small, circular, fuzzy-edged, wide-angle field. } } I'm not sure what the C-2500L looks like, but it may have a } physically larger lens that has its iris deep behind the front } surface. This might require a very long focus eyepiece, like a } 40mm Plossl, conceivably, or one with even greater eye relief, to } accomplish the optical hook-up more favorably, and may limit the } operation of the overall system to somewhat lower magnifications. } Oh, once you have a compatible eyepiece, then you can use Barlow } lenses to get back needed magnification for your desired image } scale. The only probmen there is the setup gets pretty long as } these lenses are stacked up, and stability may suffer. } } The Nikon, as mentioned, has been found by many to be almost } ideal for afocal photography of the moon and planets. } } Chuck }
************************************************************************* {/bigger} {/bold} Interest in the sophisticated fluorescence imaging techniques of Fluorescence Resonance Energy Transfer (FRET) and Fluorescent Lifetime Imaging Microscopy (FLIM) amongst the biological research community has grown in recent years. FRET imaging provides a tool to solve complex structural associations at resolution limits beyond conventional optical imaging. FLIM allows the measurement of FRET without the significant problems associated with intensity based FRET measurement, as well as faster, more accurate and quantitative measurement of cell physiology. These techniques are also being implemented in high-throughput screening regimes for drug discovery. Invited lectures by a distinguished group of scientists will concentrate on new technical developments in these areas and demonstrate successful application of these techniques in biological and industrial settings.
A poster session has been organized for June 9th so that registrants may present their experiences with FRET and FLIM.
We anticipate that this will be a most enjoyable as well as intellectually stimulating symposium.
************************************************************************* {/bigger} {/bold} Interest in the sophisticated fluorescence imaging techniques of Fluorescence Resonance Energy Transfer (FRET) and Fluorescent Lifetime Imaging Microscopy (FLIM) amongst the biological research community has grown in recent years. FRET imaging provides a tool to solve complex structural associations at resolution limits beyond conventional optical imaging. FLIM allows the measurement of FRET without the significant problems associated with intensity based FRET measurement, as well as faster, more accurate and quantitative measurement of cell physiology. These techniques are also being implemented in high-throughput screening regimes for drug discovery. Invited lectures by a distinguished group of scientists will concentrate on new technical developments in these areas and demonstrate successful application of these techniques in biological and industrial settings.
A poster session has been organized for June 9th so that registrants may present their experiences with FRET and FLIM.
We anticipate that this will be a most enjoyable as well as intellectually stimulating symposium.
} Dear colleagues, I would like to buy a digital camera } for my Axiolab Zeiss microscope. } Unfortunately I am a bit confused in the amount of } available data. I would need a digital camera of the } resolution that matches the quality of film cameras } in order I need not scan photos or negatives. } ...
To give you an idea of what you are asking: For comparable resolution, the camera would need deliver more than 6M pixels ... and there is also the question of a digital camera capturing the gamut of color capable of film. For example, the camera you mention, which is aimed at consumers, probably delivers a gamut aimed at the "sRGB" color space. Only a film scanner can capture a color gamut comparable to "Ektaspace RGB". Still, your camera is likely to do a very good job if properly adapted to the microscope. You will need a 1X C-mount adapter for the microscope head, and an adapter for mounting the camera on the C-mount. These are readily obtained for Nikon Coolpix cameras, possibly yours too.
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - mshaf-at-darkwing.uoregon.edu Geological Science's Electron Probe Facility - University of Oregon http://epmalab.uoregon.edu/
Hello Tina, I have the scanner you are looking at & like it a lot. To be quite honest I do not find that I need to exploit it'll full capably. If I were in the market again, looking at newer technology I would be interested in a faster scanner of similar quality. Yes I want my cake & to eat it too :). I'll give you this analogy. If I have 10 negatives I will franchise my time, that is let things scan while I hang out in the office doing other things. If I have 20 negatives, I'll probably goto the darkroom to make photos. It is quicker & paper is cheaper. BTW I have an Epson 870 inkjet that produces nice quality images... cost is down to $180 US, (now the Epson 880)....no financial interest in these companies.
Oh yea, get the fastest computer you can afford.
good luck, Bruce Brinson Rice U.
Tina Carvalho wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, All- } } A colleague has asked for recommendations for setting up a digital } darkroom (fun to spend someone else's money!). This person would benefit } from a really good scanner that could deal with prints, large format } negatives (4"x5", 3.25"x4") as well as 35 mm slides. At one time I looked } into an Agfa Duoscan T2500. Do any of you have an opinion about this or } other suitable scanners? } } I know this subject comes up regularly, but I don't feel bad about } introducing it again, since technology evolves so quickly! } } Mahalo, } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
Tina, Have your colleague check out the Imacon Flextight Precision II scanner. The optical resolution is 5760 dpi for slide-sized objects; I believe it drops to 4800 dpi for objects the size of her larger negatives. The scanner collects 14 bits of usable data per channel, which can be exported as a two bytes per channel, and has a dynamic range of 3.9 OD units (4.1 OD max). The machine is also very fast. The URL is:
At 05:22 PM 4/4/2001 -1000, Tina Carvalho wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Tina,
I have the Duoscan and a Nikon slide scanner. The Duoscan can scan slides on the special tray feature but side by side comparisons of the Duoscan and Nikon show that the Nikon scan is much better. For the larger negs we had a special tray made for the Duoscan and we scan in our EM negs. The Nikon has gotten much cheaper and an excellent scanner can be had for $700 with Digital ICE, something you want.
Get two scanners.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
I forgot to mention John that we do not use formar coated grids. For better assessment of the renal biopsy we do not mince the sample, either. Please see our Web page for more details. {pathology.lsuhsc.edu/Pathist/dx_home.htlm} click on M.diagnostic service and then on renal biopsy. Our diagnostic em lab continues to send poloroid HRLM pictures and TEM B&W contact prints with each report.
Lehigh University seeks an Electron Microscopy Technician to perform duties in support of the Microscopy Center of the Materials Science and Engineering Department. The person appointed will work with other technical staff to instruct students in the operation of microscopes and other equipment; maintain and repair instruments; carry out upkeep of the lab; support research professors and students; analyze samples; give tours and demonstrations; maintain a safe environment and perform other assigned duties. A bachelor's degree in physical science and/or 4+ years related work experience is required. Candidates should be familiar with electron microscopes, mechanical and electronic equipment, vacuum systems, computers (PC and/or Mac) and EDS/WDS systems. Experience with a microprobe would be especially valuable. Good communication and interpersonal skills are essential.
Lehigh University offers excellent benefits including medical, vision and tuition. Interested candidates should forward their resume to Jennifer Mohney, Human Resources, 428 Brodhead Avenue, Lehigh University, Bethlehem PA 18015. EEO/AA
-- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
One of our students is trying to stain the DNA of osteoblasts using bisBenzamid, Hoechst no. 33258 trihydrochloride. She is finding conflicting information about the concentration to use and the lethal dose.
I would appreciate it if someone can provide or point us to a protocol they have used successfully. We are primarily a materials lab and do not have a lot of biological reference material at hand.
One of our students is trying to stain the DNA of osteoblasts using bisBenzamid, Hoechst no. 33258 trihydrochloride. She is finding conflicting information about the concentration to use and the lethal dose.
I would appreciate it if someone can provide or point us to a protocol they have used successfully. We are primarily a materials lab and do not have a lot of biological reference material at hand.
I was initially hesitant to introduce a subject that gets periodically posted here, but received so many enthusiastic messages about print and negative scanners that I thought I'd continue the thought. I will be happy to summarize the responses and throw in some opinions of my own. As more and more of us convert to "digital darkrooms" we can share more of our experiences with hardware and software.
I am helping a former traditional photographic media user/computerphobe set up digital imaging capabilities since her university/museum department is closing their darkroom facilities and reassigning personnel (sigh). She has what appears to be a decent budget (until I started pricing the good stuff!). I am proposing she get a fast (733MHz) G4 Mac with maximum (1.5GB) RAM, and she saw and fell in love with the Apple 22" cinema display (as did I when I saw it in person). People seem to like the Agfa T2500 scanner for prints and negatives. A moderate color printer, since she has access to other really good printers in her department. A Nikon Coolpix 990 digital camera for on- and off-microscope. Photoshop 6.0, for which I'll train her. Corel Draw for vector graphics?
Additions, subtractions and comments will be welcome. I'll summarize after a reasonable amount of time and we'll see if there is a consensus on the ideal digital darkroom!
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
How does your institution handle short term storage in the lab of waste osmium tetroxide. Does it need to be in a fume hood? We keep it in closed glass containers mixed with vegetable oil. Does anyone have a reference regarding whether or not it needs to be stored in a hood? Thank you for any responses.
John, I forgot to mention that we do not use formar coated grids. For better assessment of the renal biopsy we do not mince the sample, either. Please see our Web page for more details. {pathology.lsuhsc.edu/Pathist/dx_home.htlm} click on M.diagnostic service and then on renal biopsy. Our diagnostic em lab continues to send poloroid HRLM pictures and TEM B&W contact prints with each report.
I am a laboratory instructor at Brookdale Community College in Lincroft, NJ who operates a Zeiss 9C TEM which was donated to the college. We are still in the process of being able to make our own grids. Does anyone have some grids that they would be willing to sell or donate to the College? Grids of anything would be greatly appreciated, but basic cellular structures is really what we are looking for as we are a community college and only have first and second year students. Thanks in advance for your help.
I used Agfa DuoScan HiD earlier and I try to get it here as well. I like that machine a lot. It's optical resolution is 1000x2000 Dynamic range is 3.7D, which would help scanning DP's. If you want more info you can have a look at:
http://www.agfa.com/scanners/duoscan_HiD.html
Printing is another task you can buy things from AGFA as well. Their photoprinter is just excellent, but a bit expensive. I have tried nice HP injet printers with great success.
Cheers! Csaba
-- ____________________________________________ Csaba Cserhati Univ.of Debrecen / Dept. of Solid State Phys. Hungary tel/fax: 36 52 316073 e-mail: cserhati-at-delfin.klte.hu ____________________________________________
I used Agfa DuoScan HiD earlier and I try to get it here as well. I like that machine a lot. It's optical resolution is 1000x2000 Dynamic range is 3.7D, which would help scanning DP's. If you want more info you can have a look at:
http://www.agfa.com/scanners/duoscan_HiD.html
Printing is another task you can buy things from AGFA as well. Their photoprinter is just excellent, but a bit expensive. I have tried nice HP injet printers with great success.
Cheers! Csaba
-- ____________________________________________ Csaba Cserhati Univ.of Debrecen / Dept. of Solid State Phys. Hungary tel/fax: 36 52 316073 e-mail: cserhati-at-delfin.klte.hu ____________________________________________
I used Agfa DuoScan HiD earlier and I try to get it here as well. I like that machine a lot. It's optical resolution is 1000x2000 Dynamic range is 3.7D, which would help scanning DP's. If you want more info you can have a look at:
http://www.agfa.com/scanners/duoscan_HiD.html
Printing is another task you can buy things from AGFA as well. Their photoprinter is just excellent, but a bit expensive. I have tried nice HP injet printers with great success.
Cheers! Csaba
-- ____________________________________________ Csaba Cserhati Univ.of Debrecen / Dept. of Solid State Phys. Hungary tel/fax: 36 52 316073 e-mail: cserhati-at-delfin.klte.hu ____________________________________________
Based on historical precedent we keep used osmium tetroxide in the fridge in a "Kilner" jar (for pickled fruit and veg.), which has a rubber seal.
BTW what ratio of vegetable oil to osmium solution do you use?
Dave
On Thu, 5 Apr 2001 20:30:30 -0500 Michele von Turkovich {mvonturk-at-zoo.uvm.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } How does your institution handle short term storage in the lab of waste } osmium tetroxide. Does it need to be in a fume hood? We keep it in closed } glass containers mixed with vegetable oil. Does anyone have a reference } regarding whether or not it needs to be stored in a hood? Thank you for any } responses. } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
In respose to Tina's post, I have not seen any mention on the list of the scanner I purchased a few weeks ago, the Epson Expression 1640XL. It has 1600dpi optical resolution (scans at a hardware resolution of 1600x3200 dpi) 42 bit color (14 bit gray) and Dmax of 3.6. It is large format, and the transparency adapter comes with a range of negative holders. Has SCSI or USB interfaces with firewire as an optional extra (I use USB on a Win 2000 system). Of course, you pay for what you get - it isn't cheap.
We are only just beginning to learn how best to use all the resolution and bit depth we now have, but I and my users love it!
This is not a comparison, of course (I haven't used the other models) but just to say we are happy with what we have.
Tony.
} } Hi, All- } } A colleague has asked for recommendations for setting up a digital } darkroom (fun to spend someone else's money!). This person would benefit } from a really good scanner that could deal with prints, large format } negatives (4"x5", 3.25"x4") as well as 35 mm slides. At one time I looked } into an Agfa Duoscan T2500. Do any of you have an opinion about this or } other suitable scanners? } } I know this subject comes up regularly, but I don't feel bad about } introducing it again, since technology evolves so quickly! } } Mahalo, } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** }
Hysterical precedents aside. The idea of the oil is to absorb all tetroxide vapours and render the substance harmless. Metallic Os is essentially non-toxic, so the treated tetroxide waste should smell of whatever vegetable oil and not emit any of the musky smell emitted by osmium tetroxide. Neither should the treated material be regarded as hazardous waste - but I expect that no safety officer would have the courage to make that declaration. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, April 06, 2001 6:52 PM, Patton, David [SMTP:David.Patton-at-uwe.ac.uk] wrote: } } } } Based on historical precedent we keep used osmium tetroxide } in the fridge in a "Kilner" jar (for pickled fruit and } veg.), which has a rubber seal. } } BTW what ratio of vegetable oil to osmium solution do you } use? } } Dave } } } On Thu, 5 Apr 2001 20:30:30 -0500 Michele von Turkovich } {mvonturk-at-zoo.uvm.edu} wrote: } } } } } } } How does your institution handle short term storage in the lab of waste } } osmium tetroxide. Does it need to be in a fume hood? We keep it in closed } } glass containers mixed with vegetable oil. Does anyone have a reference } } regarding whether or not it needs to be stored in a hood? Thank you for any } } responses. } } } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } } }
between membranes in all cellular components that have two or more membrances and
also seeking "volume and size" of all known subcellular organelles with or without membranes. also seeking " other physical measurement parameters" of cellular components by cell type.. by cell age.. etc. thanks.
Computer Aided Cell and Molecular Biology (CACMB), not medicine, will find the cure for cancer and other diseases. There will always be a need for the trained clinician (MD/RN) but, advanced diagnostic and treatment option selection has become gene based, has moved from the physician's practice to the computerized cell and molecular biology laboratory, and appropriate treatment options should now be based on the personal biology of the patient.
Adobe Illustrator or Freehand for vector graphics. Corel Draw is less common and has a proprietary file format that has caused me troubles preparing articles for Microscopy Today. Corel can save in other formats, but people have to use that option. Also, Adobe has educational pricing, and special package prices that bundle full versions of Photoshop and Illustrator.
Phil
} Hi, again- } } I was initially hesitant to introduce a subject that gets periodically } posted here, but received so many enthusiastic messages about print and } negative scanners that I thought I'd continue the thought. I will be happy } to summarize the responses and throw in some opinions of my own. As more } and more of us convert to "digital darkrooms" we can share more of our } experiences with hardware and software. } } I am helping a former traditional photographic media user/computerphobe } set up digital imaging capabilities since her university/museum department } is closing their darkroom facilities and reassigning personnel (sigh). She } has what appears to be a decent budget (until I started pricing the good } stuff!). I am proposing she get a fast (733MHz) G4 Mac with maximum } (1.5GB) RAM, and she saw and fell in love with the Apple 22" cinema } display (as did I when I saw it in person). People seem to like } the Agfa T2500 scanner for prints and negatives. A moderate color printer, } since she has access to other really good printers in her department. A } Nikon Coolpix 990 digital camera for on- and off-microscope. Photoshop } 6.0, for which I'll train her. Corel Draw for vector graphics? } } Additions, subtractions and comments will be welcome. I'll summarize after } a reasonable amount of time and we'll see if there is a consensus on the } ideal digital darkroom! } } Mahalo, } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
We keep used osmium in small sealed bottles in the hood. Our environmentals services office would rather have **undiluted/unmixed** (no oil) waste for them to neutralize. They have no way of knowing whether all the osmium has been denatured by attachment to oil and would have to treat the whole bottle as osmium waste making for more volume to have to dispose of. Check with your hazardous waste office.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Email: inikolak-at-mred.tuc.gr or eisodos-at-otenet. Name: Irene Nikolakaki
Organization: Technical University of Crete
Education: Graduate College
Location: Chanea, Crete,Greece
Question: Dear sir/madam,
I am a postgraduate student at the Technical University of Crete and I seek information (as detailed as possible) on how enumeration of airborne microorganisms collected on Nuclepore filters is done with the use of epi-fluorescence microscopy.I would be grateful if you could provide me with this infromation or any kind of help as to where I can find it.
I both run the microscope labs here and am a researcher in materials engineering.
First I want to say that I strongly support the use of the digital laboratory. While we still occasionally use film for our highest quality requirements, in general we are fully digital. The use of digital cameras has really expanded our undergraduate teaching laboratories and has sped up our research.
I have found one "dark-side" to a digital imaging laboratory as a lab manager. As the lab manager, I have found that keeping a digital laboratory up-to-date is much more expensive than the film laboratory. When we were only film, we had to repair the film cartridges for our Polaroid PN film (it takes about five minutes) and have the microscopes cleaned about once a year at a cost of about $1k.
The digital lab. is much more expensive time and repair wise. Because we crunch our computers with our image size and storage, it takes more of my time to keep stuff going. All our computers are networked and in addition to work, the students tend to junk up the computers with downloads etc which stops them from working for the image processing work. This requires continual monitoring on my part (in spite of rules against using them for these applications!) In addition, keeping computers that will run the data is expensive. I buy pretty much the best out there, but somehow upgrades are still inevitable. I also have to supply print cartridges,etc. Researchers always supplied their own film and dark room supplies. In addition, I've had to have our cameras repaired numerous times. The cost was high (at least $500) and they stayed gone for up to a month. Finally, some of my cameras are about 3 years old. I can see a degredation in the image quality from when they were purchased. The cameras are much noisier. I see a future of regular replacement of my cameras in addition to the computer upgrades. So while the cost to the researchers is lower (which helps me as a researcher), the cost to the lab itself is higher (which hurts me as a lab manager). I'm working on setting up a fee schedule for this equipment but REALLY hate to have to do it. All of you who do this in a university know how painful it is!
Regarding the camera purchase-in addition to considering the camera resolution and cost, I think you should consider the image transfer. I recommend considering a camera with immediate transfer of the image to the microscope if you have numerous inexperienced users. Being able to focus on the screen is extremely helpful. The image transfer time is also important if you have many images to capture. We do image analysis on numerous images and some of the cameras have about a 30 second transfer time for decent resolution. This would be unbearable for the number of images we collect. I'm not sure how the Nikon Coolpix works but this should be considered by anyone that is purchasing a digital camera.
Good luck!
Robin Griffin UAB
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: Thursday, April 05, 2001 6:09 PM To: Microscopy Listserver
Hi, again-
I was initially hesitant to introduce a subject that gets periodically posted here, but received so many enthusiastic messages about print and negative scanners that I thought I'd continue the thought. I will be happy to summarize the responses and throw in some opinions of my own. As more and more of us convert to "digital darkrooms" we can share more of our experiences with hardware and software.
I am helping a former traditional photographic media user/computerphobe set up digital imaging capabilities since her university/museum department is closing their darkroom facilities and reassigning personnel (sigh). She has what appears to be a decent budget (until I started pricing the good stuff!). I am proposing she get a fast (733MHz) G4 Mac with maximum (1.5GB) RAM, and she saw and fell in love with the Apple 22" cinema display (as did I when I saw it in person). People seem to like the Agfa T2500 scanner for prints and negatives. A moderate color printer, since she has access to other really good printers in her department. A Nikon Coolpix 990 digital camera for on- and off-microscope. Photoshop 6.0, for which I'll train her. Corel Draw for vector graphics?
Additions, subtractions and comments will be welcome. I'll summarize after a reasonable amount of time and we'll see if there is a consensus on the ideal digital darkroom!
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
Hello, On the September/October issue of Microscopy and Microanalysis, they have a cover picture of a yeast SEM image collected by David Scharf. I was wondering if anyone knew the details of how the sample was prepared. It looks as though the sample was processed directly from the medium it was growing on. I am used to processing bacteria and yeast by filtering through membrane filters, or depositing on polylysine treated glass/silica. I was wondering if there was something different done for this sample?
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
I am in need of some form of a conductive film coater for SEM sample preparation. I have found a Tousimis Research Corporation Samsputter-2a sputter coater, though it may not be working properly.
Until now, I have been considering constructing my own sputter coater from microwave oven parts and the like, but I have not yet found any books or other resources with clear enough information to get my confidence to the point that I feel that I can do it. I have an understanding of what is needed electrically, but I need something that fills in some details like "Be sure that the target to sample distance can be adjusted between X and Y. You need to get a QRZ type needle valve to admit the argon", etc.
Does anyone have experience with the Tousimis Samsputter 2a? From what I can see, it is an extremely simple unit. Is it something that would be worth repairing? Would it best be used to scavenge parts for a home-built unit?
Since I have breeched the question of home-built sputter coaters, does anyone know of any resources that describe the dos and donts of building one? I know that there are a number of vendors on the listserv. What are the critical features of your units that I just can't get in a home-built unit and just can't live without?
To put this all in perspective, I have an old SEM with 100 Angstrom resolution at best. I don't need flawless, grainless deposition. I have close to zero budget. Even a used sputter coater at market prices is far too expensive. Either I make something like this work, or I learn to work with uncoated samples (which will be difficult, since I am interested in looking at clays - small particle size with next to zero conductivity).
I suggest to use SEM (with calibrated magnification) if you could electroplate your sample with layer of Ni 0.1-0.3 mm thick. Sorry, I do not have a protocol for plating with Ni now, but I did it many times and it is pretty easy.
After coating with Ni you can cut your sample, mount it in epoxy or thermoset and prepare usual cross section. Ni would save the edge of steel layer and measurements will be very simple.
If surface of you samples is flat, you can cut them (better with diamond saw), mount them in epoxy with a steel strip (instead of Ni) attached to steel layer and polish. It is less dependable but still not bad method.
Vladimir Dusevich
-----Original Message----- } From: "rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com [mailto:"rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com] Sent: Thursday, April 05, 2001 8:25 AM To: microscopy-at-sparc5.microscopy.com Cc: hban-at-eng.uab.edu
We have a professor here who has a 1 cm x 2.5cm steel layer about 100 um in thickness on a ceramic substrate. The metal layer was sputter deposited onto the ceramic substrate. The ceramic substrate extends past the metal layer. He needs to get a thickness gradient across the steel layer along the 2.5 cm length. He would like to have one end at about 50 microns in thickness and the other at 2 microns with a gradually decreasing thickness gradient. Steps down would be ok although a smooth transition would be better. We have a laser profilometer to measure anything that we produce.
Does anyone out there have any ideas how to do this? Would a tripod polisher work? I thought about electropolishing and masking off portions at a time but I worry about what will happen at the interface between the ceramic and the metal. Could we alter a dimpler?
Osmium waste is toxic and hazardous due to its reactivity and heavy metal characteristics. One of the "urban legends" that abounds is that Osmium mixed with oil is "neutralized" and made safe. It is true that much of it is reduced to metallic Osmium. However in a bizarre incident it was shown in our lab years ago that reduced Osmium is very reactive with strong oxidizers. In the incident some reduced Osmium was accidentally spilled in a sink which had a small amount of Hydrogen Peroxide in the drain. The ensuing yellow gas cloud of newly formed Osmium Tetroxide from the exothermic reaction was very toxic!!!! I mention this only to confirm that Sara Miller is right again as usual and that mixing Osmium waste with other compounds does not make it safe and does increase the volume.
Hi All, I've been tearing hair out all day trying to find a recipe for GMA. In the past, I have used either JB-4 or Historesin (or Technovit) embedding kits, which have clear directions for embedding tissue and polymerizing these resins without UV. I recently ordered a straight GMA embedding kit (because it was much cheaper), and received not one bit of direction as to how I should combine the three components. I did find a protocol on EMS' website for UV polymerization, but would much prefer to use the non-UV method, since it allows tissue to be better oriented. Before I start experimenting, I thought I'd see if anyone out there can help. Thanks again for your help. Bald in Iowa, Kristen Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 515-294-8854 kalen-at-iastate.edu
Osmium metal is sometimes reported not to react with air, but other sources report slow reactivity of the metal with air to produce OsO4. It is interesting to note that the name osmium is derived from the Greek "osme" meaning smell, referring to the odour of the metal resulting from osmium tetroxide production at its surface. Presumably neither the finely divided metal nor the dioxide (osmium black) can be trusted not to oxidise to OsO4 in air. What makes osmium tetroxide especially hazardous is the fact that it is very volatile. Unlike OsO4 neither osmium metal nor osmium dioxide are volatile, and provided oxygen can be prevented from reaching them they are therefore relatively innocuous. Surrounding them in oil is probably therefore a good strategy, provided the mixture is still treated as toxic waste.
Dr. Chris Jeffree Inveresk Cottage 26, Carberry Road Inveresk Musselburgh Midlothian EH21 8PR Tel: +44 131 665 6062 FAX +44 131 653 6248 Mobile 07710 585 401
I think that I have missed part of this message string somewhere, but it sounds like you are starting with a uniform thickness steel layer that has been deposited on a ceramic substrate. I will then make the assumption that you are attempting to alter the thickness of the steel layer across the length of the sample to produce a 'wedge'. I am not sure that this will help, but you could try the wedge polishing technique used for TEM sample preparation, or bevel polishing. I have had success bevel polishing IC's, but not to the degree of accuracy that you require. I know that South Bay technologies makes a pretty good tripod with micrometer levels at all three corners that may help you out. The biggest problem will be in setting up to ensure a good gradient. You should probably try it a few times with 'dummy' samples to get the feel for how aggressive your polish angle is. I hope this helps. Nick Aitken
-----Original Message----- } From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] Sent: Friday, April 06, 2001 11:37 AM To: microscopy-at-sparc5.microscopy.com Cc: hban-at-eng.uab.edu
I suggest to use SEM (with calibrated magnification) if you could electroplate your sample with layer of Ni 0.1-0.3 mm thick. Sorry, I do not have a protocol for plating with Ni now, but I did it many times and it is pretty easy.
After coating with Ni you can cut your sample, mount it in epoxy or thermoset and prepare usual cross section. Ni would save the edge of steel layer and measurements will be very simple.
If surface of you samples is flat, you can cut them (better with diamond saw), mount them in epoxy with a steel strip (instead of Ni) attached to steel layer and polish. It is less dependable but still not bad method.
Vladimir Dusevich
-----Original Message----- } From: "rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com [mailto:"rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com] Sent: Thursday, April 05, 2001 8:25 AM To: microscopy-at-sparc5.microscopy.com Cc: hban-at-eng.uab.edu
We have a professor here who has a 1 cm x 2.5cm steel layer about 100 um in thickness on a ceramic substrate. The metal layer was sputter deposited onto the ceramic substrate. The ceramic substrate extends past the metal layer. He needs to get a thickness gradient across the steel layer along the 2.5 cm length. He would like to have one end at about 50 microns in thickness and the other at 2 microns with a gradually decreasing thickness gradient. Steps down would be ok although a smooth transition would be better. We have a laser profilometer to measure anything that we produce.
Does anyone out there have any ideas how to do this? Would a tripod polisher work? I thought about electropolishing and masking off portions at a time but I worry about what will happen at the interface between the ceramic and the metal. Could we alter a dimpler?
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Jim Darley wrote: ================================================== Hysterical precedents aside. The idea of the oil is to absorb all tetroxide vapours and render the substance harmless. Metallic Os is essentially non-toxic, so the treated tetroxide waste should smell of whatever vegetable oil and not emit any of the musky smell emitted by osmium tetroxide. Neither should the treated material be regarded as hazardous waste - but I expect that no safety officer would have the courage to make that declaration. ================================================== Jim might be right but there are other points of view on this:
1] I have been led to believe that the vegetable oil reduction of remaining unreduced tetroxide reduces things down to the dioxide, not the metal. The dioxide is a black colloidal solid (the metal is a lighter "bluish gray"). Osmium dioxide is itself relatively innocuous, for example, it is not "regulated" as a dangerous good when shipped, either domestically under US DOT rules or internationally under IATA rules.
2] Osmium is a non-renewable resource and as such, we should all be looking for ways to recycle such materials as opposed to taking them to landfills or in the case of osmium, incineration. I am no environmental "activist" myself, but I think we should all be thinking about such things. The possibility of recycling something however, is intimately connected with the economics of recycling vs. the cost of the purchase of new virgin material. And when the "used" osmium tetroxide containing aqueous liquid is "neutralized" in vegetable oil, for those involved in precious metals recycling, this act essentially kills the economics of recycling.
3] We have in beta testing stage right now an "osmium recycling kit". We are looking for a limited number of laboratories (for now, just in the USA) to participate in our beta testing of this kit. If you are interested in participating in this test, let me know off-line and I will send you the details.
4] In the mean time, I would offer the following advice. Consider using one of the other methods described on this listserver in the past, such as the ones involving KOH as a reducing agent. The reduced material in this state can be recycled economically, but only in large quantities. No one laboratory, in our opinion, could generate enough such material over a reasonable period of time, to make recycling and refining, even of the material is in this state, economical.
5] The one thing you don't ever want to do, at least in the USA, is to declare this as any kind of a "hazardous waste". Once something has been declared to be a hazardous waste, that designation can never (if my understanding of regulations is correct) be reversed, and it forever has to be treated as a waste, and translated, that means it is destined for eventual disposal by either landfill or incineration. I want to be very careful here, it is complicated, this is true so long as we are talking about a RCRA hazardous waste, that is, it meets a listing or characteristic definition. One environmental experts tells me the following: "The reason OsO4 (and OsO2, for that matter) are not regulated as hazardous has nothing to do with their human toxicity (or lack of); it is because osmium, from a regulatory standpoint, is not considered toxic to the environment." He also says it is OK to designate something as a hazardous waste with the intent to recycle it; it simply must be managed as a hazardous waste while it is on-site.
So these containers that are holding reduced material (from the tetroxide) for recycling must be labeled properly, and that would mean something like "osmium dioxide for recycling". I am getting into an area that is not black and white defined, and probably varies from institution to institution in the US, not to mention the variation from country to country. But the important thing is that from a regulatory point of view, what that label says ends up determining the possibilities for the ultimate fate of its contents.
We are striving to find a way to help people transfer, on their environmental "accounting sheets", a material from the column saying "incineration" or "landfill", to the "recylcing" column. And for those who worry about such things, from a legal liability standpoint, there is general recognition that if the material is recycled, there is far less legal liability associated with its disposal than if it is incinerated or sent to landfill. And at the same time, recycling keeps this most valuable nonrenewable resource in the stream of commerce and available for future generations.
Disclaimer: SPI Supplies has developed a kit for recycling osmium. It is not yet commercially available, but is in beta test stage. We also have the obviously ulterior motive of making sure there is osmium tetroxide available for future generations of EM users, otherwise there would be no place for firms like SPI Supplies.......
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
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Different people approach such subjects from rather different angles and they may all be right, but considering their own realities.
A couple of submissions approached the subject from an "absolute safety" point of view. I don't subscribe to that because the amount of tetroxide emitted from gram quantities of metal and some dioxide when surrounded by the tetroxide absorbing oil, would be miniscule. In my opinion, Os waste in vegetable oil is not a substantial hazard in the lab or when dumped. For many, particularly in the USA this is irrelevant, because of strict legal obligations.
There is also an environmental angle. Unfortunately, greatest safety regardless of cost, is at odds with good environmental practice. If a lot of material and energy is used to dispose of a low hazard material, then the total environmental cost may be very high when compared with an "acceptable risk" solution. We should not confuse the ever-increasing demands for greatest safety with "environmentally friendly" practices: they are not synonymous, but often mutually exclusive.
I was pleased to read some of Chuck's submission. Whatever the motivation, recycling of a limited resource is commendable. This has been tried before by turning the waste material back into the tetroxide. I understand that few people still practice the regeneration of osmium. One reason is that the actual fixative solution is poorly defined after reclamation from diverse fixation vehicles.
Chuck's idea of turning the material into metallic osmium is possible. Using precision electrolysis, for instance with SS electrodes at ~400volts, with the plate size determining amperage, the recovered metal could be 99.9% pure. The refiner, however, would charge for assaying and refining and this means another business would need to collect the osmium, resulting in double shipping and double mark-ups. Considering the few grams recoverable, instrument cost and maintenance, this would be a marginal business, but one that the larger labs certainly should consider. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Monday, April 09, 2001 4:45 AM, Garber, Charles A. [SMTP:cgarber-at-2spi.com] wrote: } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Jim Darley wrote: } ================================================== } Hysterical precedents aside. The idea of the oil is to absorb all tetroxide } vapours and render the substance harmless. Metallic Os is essentially } non-toxic, so the treated tetroxide waste should smell of whatever } vegetable } oil and not emit any of the musky smell emitted by osmium tetroxide. } Neither should the treated material be regarded as hazardous waste - but I } expect that no safety officer would have the courage to make that } declaration. } ================================================== } Jim might be right but there are other points of view on this: } } 1] I have been led to believe that the vegetable oil reduction of remaining } unreduced tetroxide reduces things down to the dioxide, not the metal. The } dioxide is a black colloidal solid (the metal is a lighter "bluish gray"). } Osmium dioxide is itself relatively innocuous, for example, it is not } "regulated" as a dangerous good when shipped, either domestically under US } DOT rules or internationally under IATA rules. } } 2] Osmium is a non-renewable resource and as such, we should all be looking } for ways to recycle such materials as opposed to taking them to landfills or } in the case of osmium, incineration. I am no environmental "activist" } myself, but I think we should all be thinking about such things. The } possibility of recycling something however, is intimately connected with the } economics of recycling vs. the cost of the purchase of new virgin material. } And when the "used" osmium tetroxide containing aqueous liquid is } "neutralized" in vegetable oil, for those involved in precious metals } recycling, this act essentially kills the economics of recycling. } } 3] We have in beta testing stage right now an "osmium recycling kit". We } are looking for a limited number of laboratories (for now, just in the USA) } to participate in our beta testing of this kit. If you are interested in } participating in this test, let me know off-line and I will send you the } details. } } 4] In the mean time, I would offer the following advice. Consider using } one of the other methods described on this listserver in the past, such as } the ones involving KOH as a reducing agent. The reduced material in this } state can be recycled economically, but only in large quantities. No one } laboratory, in our opinion, could generate enough such material over a } reasonable period of time, to make recycling and refining, even of the } material is in this state, economical. } } 5] The one thing you don't ever want to do, at least in the USA, is to } declare this as any kind of a "hazardous waste". Once something has been } declared to be a hazardous waste, that designation can never (if my } understanding of regulations is correct) be reversed, and it forever has to } be treated as a waste, and translated, that means it is destined for } eventual disposal by either landfill or incineration. I want to be very } careful here, it is complicated, this is true so long as we are talking } about a RCRA hazardous waste, that is, it meets a listing or characteristic } definition. One environmental experts tells me the following: "The reason } OsO4 (and OsO2, for that matter) are not regulated as hazardous has nothing } to do with their human toxicity (or lack of); it is because osmium, from a } regulatory standpoint, is not considered toxic to the environment." He } also says it is OK to designate something as a hazardous waste with the } intent to recycle it; it simply must be managed as a hazardous waste while } it is on-site. } } So these containers that are holding reduced material (from the tetroxide) } for recycling must be labeled properly, and that would mean something like } "osmium dioxide for recycling". I am getting into an area that is not black } and white defined, and probably varies from institution to institution in } the US, not to mention the variation from country to country. But the } important thing is that from a regulatory point of view, what that label } says ends up determining the possibilities for the ultimate fate of its } contents. } } We are striving to find a way to help people transfer, on their } environmental "accounting sheets", a material from the column saying } "incineration" or "landfill", to the "recylcing" column. And for those who } worry about such things, from a legal liability standpoint, there is general } recognition that if the material is recycled, there is far less legal } liability associated with its disposal than if it is incinerated or sent to } landfill. And at the same time, recycling keeps this most valuable } nonrenewable resource in the stream of commerce and available for future } generations. } } Chuck }
There is a proposal for TEM characterisation of cadmium telluride nanoparticles for morphology and size information. I would appreciate information from the list members whether
1. CdTe and related compounds are stable under electron irradiation 2. there is a accelerating voltage limit to be adhered to 3. any other precautions required to ensure microscope safety
My primary microscopy experience is with metallic alloys and ceramics. I have this impression that these compounds are low melting, unstable and likely to sputter onto the pole pieces. Kindly correct and advise me. I use a JEOL 2000 EX II top entry stage operating at 200 kV.
---- Divakar R Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research Kalpakkam 603102, India ----
Years ago, I did CdTe/CdS junction. As far as I can remember, both layers were beam sensitive and the layer CdS was more so. I could get fairly good twined CdTe structures as well as its overall morphology, but had difficulty revealing the details in CdS as the available effective observation time was much limited plus the CdS layer was very thin. There are a few ways to get around of the issue, including using a) C-coating, b) small spot size (} 2), and perhaps c) proper keV's (I was using 100keV, the max available voltage for me that time). Good luck!
Chao-Ying Ni Scientist Rodel Inc. USA
-----Original Message----- } From: Divakar R [mailto:divakar-at-igcar.ernet.in] Sent: Monday, April 09, 2001 3:58 AM To: Microscopy (E-mail)
There is a proposal for TEM characterisation of cadmium telluride nanoparticles for morphology and size information. I would appreciate information from the list members whether
1. CdTe and related compounds are stable under electron irradiation 2. there is a accelerating voltage limit to be adhered to 3. any other precautions required to ensure microscope safety
My primary microscopy experience is with metallic alloys and ceramics. I have this impression that these compounds are low melting, unstable and likely to sputter onto the pole pieces. Kindly correct and advise me. I use a JEOL 2000 EX II top entry stage operating at 200 kV.
---- Divakar R Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research Kalpakkam 603102, India ----
The tripod polisher may just work. Although your sample is larger than anything I have previously tripoded, it should work if you use a plain L-bracket. First, planarize your sample and the two back feet of the tripod polisher. Then adjust the two back feet to give you the wedge angle you desire (just slightly over 1 degree if my trigonometry for your sample is correct). Likewise, the Multiprep system from Allied High Tech should be able to accept and wedge polish a sample of this size.
If this approach does not work then you may need to use a precision lapping and polishing jig. You can either design your own or you may want to check with South Bay Technology. They make several jigs with precise angular control for polishing crystals. I don’t know however, if they will work with samples this large.
Hope this helps.
Eric Windsor
Disclaimer: I have no financial interest in either South Bay Technology or Allied High Tech Products. I am a satisfied customer of both. Also, there may be other products on the market that will work equally well for preparing this sample.
The opinion expressed is my own and not that of my employer (NIST).
Original Message:
We have a professor here who has a 1 cm x 2.5cm steel layer about 100 um in thickness on a ceramic substrate. The metal layer was sputter deposited onto the ceramic substrate. The ceramic substrate extends past the metal layer. He needs to get a thickness gradient across the steel layer along the 2.5 cm length. He would like to have one end at about 50 microns in thickness and the other at 2 microns with a gradually decreasing thickness gradient. Steps down would be ok although a smooth transition would be better. We have a laser profilometer to measure anything that we produce. Does anyone out there have any ideas how to do this? Would a tripod polisher work? I thought about electropolishing and masking off portions at a time but I worry about what will happen at the interface between the ceramic and the metal. Could we alter a dimpler?
i am interested in a rapid i.e. same day processing schedule, for diagnostic } renal bx. it must be reliable and the cutting qualities good, since we do a } lot of low mag work (250x). thanks in advance. } john
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} i am interested in a rapid i.e. same day processing schedule, for diagnostic {BR} > renal bx. it must be reliable and the cutting qualities good, since we do a {BR} > lot of low mag work (250x). thanks in advance. {BR} > john {BR} {/FONT} {/HTML}
} In respose to Tina's post, I have not seen any } mention on the list of the scanner I purchased } a few weeks ago, the Epson Expression 1640XL. } It has 1600dpi optical resolution (scans at } a hardware resolution of 1600x3200 dpi) 42 bit } color (14 bit gray) and Dmax of 3.6.
I would certainly believe the resolution and the color depth for this scanner is adequate, but if scanning TEM films is an issue, I'd seriously advise measuring the optical density of your films ... I've heard these approach OD} 4 ... which would imply you might consider the dedicated film scanners, e.g., Polaroid 45 Ultra or the new Nikon LS-8000.
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - mshaf-at-darkwing.uoregon.edu Geological Science's Electron Probe Facility - University of Oregon http://epmalab.uoregon.edu/
I have been listening to the thread on scanners. Has anyone done tests of how accurate they are in absolute terms for quantitative digitization?
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering & Center for Transportation Nanotechnology Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu http://www.ctn.northwestern.edu ------------------------------------------------------- The Other Nanotubes http://focus.aps.org/open/st12.html Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
Hello, I'm looking for companies (other than Gatan whom I've already contacted) that sell cryo-stage and cryo-prep. systems for a JEOL 5800LV scanning electron microscope. Thanks for any input! Tracey
Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
Has anyone used any of the denatured ethanols as a substitute for absolute ethanol?
We have recently run into some difficulty when needing to reorder 200 proof ethanol, which we use for dehydration and infiltration of samples (primarily many types of paper) prior to embedding in Spurrs epoxy, and for cleaning samples (non paper) and lenses of light microscopes. The chemical company selling the ethanol is insisting that we must have a liquor license before they will ship to us.
Ethanol denatured with a variety of substances is readily available and can be shipped with no licensing requirements. Our concern is that the denaturing agent will leave a detectable residue on lenses, samples, and may cause problems with the polymerization of Spurrs. Rather than obtaining a liquor license, we are considering using one of the 100:5 ethanol: methanol blends. If any of you have had successful or unsuccessful experiences substituting denatured ethanol for absolute in embedding or cleaning protocols, I would appreciate hearing from you.
Teresa Boes Hewlett-Packard Analytical and Development Lab 1000 Circle Blvd Corvallis, OR 97330 541-715-7055 teresa_boes-at-hp.com
Sorry about being late with this thread...Out here we do about 750 renal biopsies last year and all our sections are mounted on 300 mesh uncoated thin bar Gilder grids. We have a Philips 208S at 80kV. We can get a majority of the glomerulus to lie in the grid square to be viewed. Most of our images are shout between 2800X and 14,000X.
Last November we finally obtained the AMT Advantage HR 1K x 1K system and use it exclusively for our EM images... We also do some Neuropathology and Surgical Pathology EM in this lab.
Out here we have rigged up a system that all the Pathologists who need the EM images are setup on a EM users group on the network. I them upload the images from the computer here in the scope room to a directory on our network so the Pathologists can view the images right at their desktop. The renal Pathologist here is thrilled with the system... It cuts down our expenses, and it shortens our turnaround time on specimens from 5 days to about 3 days or less....
Eric A. Rosen Electron Microscopist UCLA Medical Center
==============================
} John: } We do around 500 renal biopsies per year and all the sections are } mounted on 200 mesh uncoated copper grids. We have an 8 year old Hitachi } 7100 and use 60kv. The majority of the glomerulus can be viewed with the } 3-4 serial sections lying randomly across the grid bars. We do not need a } picture of the whole glomerulus, rather most pictures are between 3,000 and } 10,000X. } Dr. Tibor Nadasdy is the renal pathologist and decided last year that } all our renal biopsies would be captured with the digital camera onto a } computer and sent up to him via a network to his computer. So, at the } present time we use very little EM film. He diagnoses each biopsy and } e-mails representative digitized images to the nephrologists. } } } } } Karen L. Jensen, M.S. } Project Manager & Associate Scientist } Electron Microscopy Research Core } -----Original Message----- } } From: "JHoffpa464-at-aol.com"-at-sparc5.microscopy.com } [mailto:"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com] } Sent: Friday, March 30, 2001 2:20 PM } To: microscopy-at-sparc5.microscopy.com } Subject: renal Em } ok taking a little survey. i am in a diagnostic EM lab. we mount out } sections } on formvar coated slotted grids, so he can shoot pics of the whole } glomerlus. } ok my question. how may of you out there doing diagnostis EM on renals do } this? } john
Tracey VG Microtech make Polaron integrated column-mounted cryo systems including a new system designed for high-resolution work with FEGSEMs Emitech have an off-microscope cryo-prep and transfer system suitable for LTSEM with a conventional tungsten filament or LaB6 SEM BalTec make various cryo transfer and preparation systems which could be used to transfer specimens from e.g. a freeze-etcher to SEM
see http://www.kaker.com/mvd/list.html for addresses and contact numbers for these companies Chris
----- Original Message ----- } From: "Tracey M. Pepper" {tpepper-at-iastate.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, April 09, 2001 6:52 PM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for a 2" and/or 3" scribe tool for GaN. Does anyone have any experience with a quality new/used equipment vendor who supplied such a tool in the past. I am looking for a fairly new programmable scriber.
Eric, I think that your geometry is off. The Tripod Polisher is about 50 mm long from the sample to the feet and has 1 um divisions. The arctan of 1/50000 gives about 0.001 degrees. This job needs an angle given by the arctan of 48/25000 or 0.11 degrees. Plus you need to stick the thickness at one end at a particular thickness -2 um. It is doable with the TP, but will be difficult. You are correct that they will need to planarize the sample. What I would do is also planarize the holder and mount the parallel-sided sample and then set the height of the feet to the thickness of the sample and then add the amount for the angle. Then I would slowly polish using a low value grit 1 or 3 (perhaps even 1/2) um until I went through to the substrate at one end. You would have your angle and thickness values that went from zero to the desired value and a little thicker. The trick is stopping at the right place and accounting for the wear on the feet. You should be able to watch the facet move towards one end. You can watch the progress using a glass plate and look at the thickness fringes at the facet caused by the unpolished and polished surfaces.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Eric Windsor [mailto:Eric.Windsor-at-nist.gov] } Sent: Monday, April 09, 2001 9:15 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Preparation of a steel wedge on a ceramic substrate } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------- } ---------. } } } Robin, } } The tripod polisher may just work. Although your sample is } larger than } anything I have previously tripoded, it should work if you use a plain } L-bracket. First, planarize your sample and the two back feet of the } tripod polisher. Then adjust the two back feet to give you } the wedge angle } you desire (just slightly over 1 degree if my trigonometry } for your sample } is correct). Likewise, the Multiprep system from Allied High } Tech should } be able to accept and wedge polish a sample of this size. } } If this approach does not work then you may need to use a } precision lapping } and polishing jig. You can either design your own or you may } want to check } with South Bay Technology. They make several jigs with } precise angular } control for polishing crystals. I don't know however, if they } will work } with samples this large. } } Hope this helps. } } Eric Windsor } } Disclaimer: I have no financial interest in either South Bay } Technology or } Allied High Tech Products. I am a satisfied customer of } both. Also, there } may be other products on the market that will work equally well for } preparing this sample. } } The opinion expressed is my own and not that of my employer (NIST). } } Original Message: } } We have a professor here who has a 1 cm x 2.5cm steel layer } about 100 um in } thickness on a ceramic substrate. The metal layer was sputter } deposited } onto the ceramic substrate. The ceramic substrate extends } past the metal } layer. He needs to get a thickness gradient across the steel } layer along } the 2.5 cm length. He would like to have one end at about 50 } microns in } thickness and the other at 2 microns with a gradually } decreasing thickness } gradient. Steps down would be ok although a smooth transition } would be } better. We have a laser profilometer to measure anything that } we produce. } Does anyone out there have any ideas how to do this? Would a tripod } polisher work? I thought about electropolishing and masking } off portions at } a time but I worry about what will happen at the interface } between the } ceramic and the metal. Could we alter a dimpler? } } } Thanks, } Robin Griffin } UAB } } }
How you can get the licence if you do not SELL the alcohol? Licence is only for selling..
} We have recently run into some difficulty when needing to reorder 200 proof } ethanol, which we use for dehydration and infiltration of samples (primarily } many types of paper) prior to embedding in Spurrs epoxy, and for cleaning } samples (non paper) and lenses of light microscopes. The chemical company } selling the ethanol is insisting that we must have a liquor license before } they will ship to us.
Does anybody have experience (good or bad) with Imaging Plate scanners on TEMs?
PLease reply on or off-line, I will post a summary - preserving anonymity if necessary!
thanks
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 or 6125 8525 http://www.anu.edu.au/EMU
Your point is well taken, but in this environment, there is no distinction between purchase, application or consumption.
I'm in California and have not run into this situation as yet. But I would bet that the same myopic rules would apply. I buy ethyl alcohol from Ted Pella in Redding, CA. But I don't know the specific proof of the alcohol (Cat.# 19207). It comes in 200mL bottles and poses no problem when purchased in lots of six or fewer bottles.
gg
At 03:38 PM 4/9/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
How do you define "quantitative digitization?" i.e., what variables are you dealing with in this respect? What are the "absolute terms?"
Anyone else have some ideas about this topic?
gg
At 10:10 AM 4/9/2001, you wrote:
} I have been listening to the thread on scanners. Has anyone done } tests of how accurate they are in absolute terms for quantitative } digitization? } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering & } Center for Transportation Nanotechnology } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu } http://www.numis.nwu.edu http://www.ctn.northwestern.edu
I am a Laboratory Technician currently studying for a Diploma in Occupational Health and Safety. My last study module is an Action Research project which I would like to do on the development of ergonomic microscopes and although there is a lot of information on the problems involved in microscopy and methods to relieve them I was wondering if any research has been done into the effectiveness of ergonomic design for improving microscopy diagnosis, and where I could get any papers on the subject. Thanks. Andy.
Perhaps the reason there was no recipe for the GMA is that you can vary the component ratios alot depending on the tissue you're embedding. There are several reviews from the 60s and 70s on different GMA recipes, at least for embedding plant material. One "standard" mix for plant material is 95% v/v pure GMA, 5% v/v polyethylene glycol, 0.15-1.0% w/v benzoyl peroxide (catalyst). With more catalyst, polymerisation is faster and blocks are harder, but too much produces brittle, bubbly blocks. PEG can be 0-10%, PEG 200 and 400 are commonly used.
Like other methacrylate resins, GMA can be heat polymerised but you need to exclude oxygen. You can either seal the GMA blocks in capsules - gelatin capsules for example - dent the cap to exclude as much air as possible, or cover flat embedding moulds so that they are completely sealed - e.g. use a plastic vial cap as flat mould with another on top to seal. If you have access to a vacuum oven, you can polymerise open moulds under nitrogen.
Time to polymerise depends enormously on the composition of the resin, try 60C overnight for starters if using a fairly "standard" mixture.
good luck, cheers, Rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Just a minor correction / update. VG Microtech recently changed name to Thermo VG Scientific, address and contact details remain the same, except for the surface science website:
The Polaron range continues to be represented in the US by Energy Beam Sciences.
Best regards,
Mike Wombwell Polaron Range Business Manager Thermo VG Scientific The Birches Industrial Estate Imberhorne Lane East Grinstead West sussex RH19 1UB UK Tel: +44(0)1342310296 (direct line) Tel: +44(0)1342327211 (Switchboard) Fax: +44(0)1342315074 email: mike.wombwell-at-scientific.com Website: http://www.polaron-range.com
E & OE
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: 09 April 2001 22:04 To: Tracey M. Pepper Cc: microscopy-at-sparc5.microscopy.com
Tracey VG Microtech make Polaron integrated column-mounted cryo systems including a new system designed for high-resolution work with FEGSEMs Emitech have an off-microscope cryo-prep and transfer system suitable for LTSEM with a conventional tungsten filament or LaB6 SEM BalTec make various cryo transfer and preparation systems which could be used to transfer specimens from e.g. a freeze-etcher to SEM
see http://www.kaker.com/mvd/list.html for addresses and contact numbers for these companies Chris
----- Original Message ----- } From: "Tracey M. Pepper" {tpepper-at-iastate.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, April 09, 2001 6:52 PM
Richard: Not true in the good old US of A! 200 proof EtOH is considered a controlled substance, and purchase of even small quantities is regulated. In the Histology and EM Labs here, we use a fairly large supply, so there is a company license which allows purchase of a specific amount, and that means if demands go up dramatically there is a real issue of obtaining adequate supplies from time to time. I think that the issue may well have to do with the quantities being purchased. If one buys the odd bottle now and then, there is no hassle. But when usage passes a certain point, then licenses are necessary. Keeps all of the bureaucrats in jobs. As for the license, Teresa, it is mostly just paperwork. Requires you to specify purpose for use, amounts that will be required, etc. If I remember correctly (had to do this myself a _very_ long time ago--now an internal supply room clerk does it), this is an annual process. And, like most labs, since you have to account for every drop of reagent that enters and leaves the lab for EPA, state EPA, OSHA, etc, you will likewise enter the EtOH into that stream, thus showing that you aren't using it for more pleasurable ends.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals NB--personal opinions and experiences only expressed above. On Tue, 10 Apr 2001 08:38:10 +1000, Vr. Richard Bejsak-Colloredo-Mansfeld wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | How you can get the licence if you do not SELL the alcohol? | Licence is only for selling.. | | } We have recently run into some difficulty when needing to reorder 200 | proof | } ethanol, which we use for dehydration and infiltration of samples | (primarily | } many types of paper) prior to embedding in Spurrs epoxy, and for cleaning | } samples (non paper) and lenses of light microscopes. The chemical company | } selling the ethanol is insisting that we must have a liquor license before | } they will ship to us. | | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
I am interested in freeze drying pieces of murine liver tissue for subsequent embedment in Spurr's epoxy resin. Does anyone have experience with this technique? Specifically I would be interested in times and temperatures. I will be using a turbo freeze dryer from EMS to freeze dry the tissue.
Thanks in advance,
Germaine G. Boucher TEM Lab Pfizer Global Research and Development Groton, CT
Would noise be a good criterion? Say, for a perfectly evenly darkened film (if such a thing existed, or at least even on a scale { { collected pixel size) - what is the value of the noise (standard deviation of pixel value) as a function of film darkness (density)?
This would presumably improve with the time of collection. Thus how "good" your scanner is depends on how you run it or whether it lets you take a slower scan or to average multiple scans. With the exception of drum scanners these devices all use CCD arrays. So what is probably most of interest is the signal to noise ratio as a function of illumination intensity, with everything known about CCD's going into determining this. The maximum density the scanner can handle is just the point at which the noise swamps the signal.
There must be some good literature out there on the sources of noise, optimizing collection (scan) time etc. One article which might be a starting point is:
G. H. Campbell, W. E. King and D. Cohen "Analysis of Experimental Error in High Resolution Electron Micrographs", Microscopy and Microanalysis vol. 3 (1997) p. 451.
This is not very detailed, and treats only the total random noise, i.e. grouping noise arising in collecting the image with that arising from the scanner.
Now, finding a good "Consumer Report" test with hard numbers on commercial models is likely to be a lot harder!
Wharton
} -----Original Message----- } From: Gary Gaugler [SMTP:gary-at-gaugler.com] } Sent: Monday, April 09, 2001 10:37 PM } To: L. D. Marks } Cc: MSA listserver } Subject: Re: Scanners: quantitative accuracy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } How do you define "quantitative digitization?" i.e., what } variables are you dealing with in this respect? What are } the "absolute terms?" } } Anyone else have some ideas about this topic? } } gg } } } At 10:10 AM 4/9/2001, you wrote: } } } I have been listening to the thread on scanners. Has anyone done } } tests of how accurate they are in absolute terms for quantitative } } digitization? } } } } ------------------------------------------------------- } } Laurence Marks } } Department of Materials Science and Engineering & } } Center for Transportation Nanotechnology } } Northwestern University } } Tel: (847) 491-3996 Fax: (847) 491-7820 } } mailto:ldm-at-risc4.numis.nwu.edu } } http://www.numis.nwu.edu http://www.ctn.northwestern.edu }
To the person having trouble getting 200 proof ethanol:
If you are embedding in Spurrs, you do not need ethanol or P.O.. Just dehydrate in E M grade acetone and infiltrate with acetone/Spurrs and you will get good results. Keep it simple, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
I have put together a summary of responses I got from my inquiry about a week or so ago about EDS systems. I really appreciate everyone's input. We haven't made any decisions yet since we are still gathering information but your responses will help. I've included the responses in their entirety so I hope this helps others as well.
Thanks again from everyone who contributed.
Jean Ross Central Microscopy Research Facility University of Iowa
----------------------------------------------------------------------------- I have been using an IXRF EDS / Gresham Detector (with IXRF digital pulse processor) system for almost 2 years. Initially, the IXRF was installed on an ETEC. About 15 months ago the Etec was replaced with a Hitachi 3500, a new detector (Gresham) was purchased, and the EDS was installed on the 3500. Generally, I am quite satisfied. There are minor software bugs, but IXRF has been reasonably good at fixing them when discovered. It has been my experience that all the systems have bugs, perhaps some more than others. Prior to the IXRF, I had a Kevex 8000/Delta.
Low end noise and broad peaks were evident on first installation, but were soon fixed by tweaking the detector preamp and pulse processor amp.
I am still running their first software package, "Iridium". I have the newest release, "EDS 2000", but lack of time has kept me from installing and checking it out.
I should mention that IXRF is a "virtual" company, with people spread out between Texas, California, etc. This has not proven to be a problem.
Woody White McDermott Technology, Inc. nwwhite-at-mcdermott.com
We have had EDAX for about a decade and a half, and we are very pleased with the product and the service we get. Carol Heckman heckman-at-bgnet.bgsu.edu
I would recommend you give consideration to Doug Connors at TN Analyzer Service, Inc. of Dane, WI. Doug has rebuild and upgraded detectors for me for the last 6 years. He is dependable knowledgeable, and economical as well.
Bob Roberts EM Lab Services, Inc. 2409 S. Rural Rd Suite C Tempe, Arizona 85282 (480) 967-3946 bobrobs-at-earthlink.net
We recently purchased a Noran Instruments Vantage DS1. We purchased this based on some very impressive demonstrations of software that the salesperson brought in. Unfortunately they are still working out the bugs in their software. Everything they have is ported over from Unix, and literally runs in a unix shell on an Microsoft Windows NT platform. This makes their software fairly buggy. Their response time to fix major bugs and hang-ups in the software has been very slow, and if given the opportunity to do it all over again I'd probably look at Oxford Instruments. I would still rate the quality of the equipment very high. Our detector performs at the specified resolution, and is a good piece of equipment. Now if they could only get the software end of it straight... My vote: 1) Oxford Instruments 2) Noran Instruments 3) Edax or some of the smaller players The benefit with going with a larger company is support and upgrades. We have a 10 year old WDX that we just purchased new software and interface for last year. Our old EDS was given some trade in value by Noran. And we all know how valuable a service contract can be...
Get back to me if you have any more questions, ~Jonathan Jonathan Dunlap Analytical Laboratory Manager Osram Sylvania Inc. 816 Lexington Avenue Warren, PA 16365 Ph: 814-726-6991 Fax: 814-726-6942 Jonathan.Dunlap-at-sylvania.com
We have an Oxford Instruments Link ISIS Model 200 on our 2460N. We have been happy with it, but I don't know about the direction that Oxford is heading. I don't care for the feel of their new INCA software. Some might like it. It also seems to be slow coming together. Some of the functions are still lacking after 2 (or is it 3) years of seeing it at MSA.
We have an IXRF system on our JEOL 840A. It was a good price ($30K) for an upgrade to our Kevex several years ago. It does what we need. They keep at work on the software and have it freely available on the web. I might have to pay closer attention and stay away from the beta stuff. They are still working on it. They also have a nice digital pulse processor which stills stand alone for about $5k.
I still feel funny about some contacts with EVEX. I can't say much about EDAX, NORAN, or PGT. They should all have good stuff but it might be pricey. The last we seriously looked at them was 6 years ago or so when we opted for the Oxford.
I was intrigued by the unit from Quartz PCI. I think it was called X-ray One, or such. It was new at MSA 1-1/2 years ago but looked promising.
We purchased an EDAX Falcon system for our Hitachi S-3000N and I've been pleased with it. It has better light element sensitivity than most which was very important to me although I don't think that its mapping capabilities are as good as PGT's, say. I don't have direct experience with Noran although I did talk to them and their system seemed ok - but logistics didn't favor Noran so I passed on them. EDAX does have good integration with the Hitachi and the Quartz database.
I'd be glad to respond more specifically if you'd like.
Richard Shalvoy Arch Chemicals Cheshire, CT RBShalvoy-at-archchemicals.com
I have an iXRF systems out of Texas using a Gresham detector. It works well. Not the most cutting edge, but they are one of the "start ups". They have been around for I guess 6-7 years. I have a digital pulse processor and completely active control for x-ray maps and such. They are very price competitive, but lack a dedicated technical support person. You talk to the programmer or electronics expert, but no techs on the phone whenever you have a software question. But, if you willing to wait a day for some answers then they are worth it. I haven't run across the problem where I thought, "if I just had a better system". If you want to integrated w/ WDS than maybe Noran. Also, if you want to integrated w/ motorized stage control, I don't think they off such a package, like the bigger companies.
I have a Hitachi 450. I used to run a 2400 and 500 before I quit my day job and went out on my own. I am very happy w/ Hitachi.
I use Oxford ISIS300 system on HITACHI S-3500N (with VP mode) for light element analysis, mostly C, O, N, F, P, S, Si, Mg, as well as metal Co, Ni-P, Pt, Cr, Fe, W, etc. This system works well. One useful function is the overlay of 2 spectrums. I can easily subtract the blank from the sample spot and make it easy to identify what is (are) in the sample. I am sure some other program may have this kind of function, but I have not seen.
Zhiyu Wang zhiyuw-at-home.com I would be interested in seeing the responses as I am going to try and get funding next year for a replacement for our EDAX PV9100 on an Hitachi s-450.
We have been running EDX on SEMs and TEMs for many years. We used to have a range of systems from Kevex, PGT, Noran, Link, EDAX, however a few years ago we decided that we ought to standardize on one common system. After evaluation we bought three Oxford Instruments ISIS systems. Whenever we have upgraded or bought new systems they have been Oxford Instruments ISIS or now INCA.
I have been happy with the ISIS except for the file handling that was not designed for a multi user facility such as ours (approx 120 EM users in total roughly 25 to 30 swapping every year). I am really quite impressed by the INCA, Oxford Instruments are, at last, listening to the users and adding user requested facilities. They have sorted out the file handling mess of the ISIS and structured it well for an SEM user (not quite as well for a TEM user but there are less of us). The software structure is quite intuitive and there is a really impressive help menu and explanation of everything from the physics of X-ray generation, how EM’s work, how detectors work and how to analyze samples.
Their detectors have always been good and the SATW (thin window detectors) still have a reasonable efficiency at low Z. B is possible but C is easy and even the N peak is over 30% efficient (there is often a high absorption at N).
Another feature that is invaluable for TEM is the integral shutter that will close when the count rate is too high. This protects the crystal, it prevents it overloading and shutting down or worse the crystal efficiency may change for a few minutes until it recovers fully. This may affects your quantitative work. In TEM this is usually caused by hitting the grid bar and not really a problem in SEM but I don't know what secondaries and ions you will have in a variable pressure SEM. It could be useful for you, check with other high pressure SEM users.
Regards, Ron
Please note: Oxford Instruments have upgraded an ISIS to an INCA system in my department, without charge, in return for access to the instrument for development projects and demonstrations for a fixed number of days. I receive no benefit from this and the department has no benefit from Oxford Instruments sales. I remain a thorn in the flesh of all our suppliers if I think they could improve their products or service. ron.doole-at-materials.oxford.ac.uk
Hello, I am very familiar with the Oxford ISIS 300 series spectrometers. They are ok, and the new Inca system looks good too. However, I recently saw the PGT spectrometer at Lehigh and it is very impressive.
I'm also in the market for an EDS system and have looked at EDAX, Noran, PGT and Oxford.
I edited out PGT because in order to quantify you have to optimize the system for the type of sample by playing with fudge factors, which none of the other systems have to do (though one of them, I think Noran, lets you adjust a sensitivity factor if you want to, but they didn't do it on my samples that were tested against known microprobe results and the answers were fine). I also eliminated Oxford, though it has a terrific user interface (maybe at the expense of functionality), because they consistently IDed my aluminum peaks as Br or Tm (!); this made me wonder about all their algorithms. They claim it had to do with the takeoff angle on the particular SEM being used, but that shouldn't be a factor.
I like both EDAX and Noran, though for different reasons. EDAX user interface is better than Noran's, though again, I think Noran possibly offers more routines (it's hard to keep track and see absolutely everything a system has to offer in a demo day....).Noran can multitask - work on several programs while a spectral map is being collected, for instance (does EDAX? I have to check). But EDAX has a beam skirt reduction routine for low vac mode (though it's time consuming, so a bit cumbersome), and their peak modeling is right up front - but Noran can put theirs up front also if you want to have it accessible (yes) and I think Noran might be a little better engineered.
As you can see I'm still in a quandary (ditto for the two contending SEMs, LEO and ESEM). Whatever I decide I'll still be very interested in the results of your posting - especially if other folks' info comes in within the next week or so it would help in my decision too.
I hope my input helps a little. Good luck with your quest!
I looked into Noran, EDAX, and PGT. Noran was quickly culled (less user friendly, less abilities, didn't work right during demo), but EDAX and PGT both seemed to have equivalent capabilities (PGT claimed a 'proprietary' signal amplifier/digitizer doohickey, but it was only in the placement.) For the long-time spectrum gathering (I forget the technical term), PGT makes many passes with short dwell times while EDAX dwells on each pixel much longer to collect data & does it in 1 pass. Kinda 6 of one, half dozen of the other. What made us choose the EDAX Phoenix system was the fact that the PGT software was UNIX-based (although hidden) while EDAX is PC-based. I've heard rumors that PGT is switching to PC-based; we purchased our system in 1999. I've also heard that Noran has practically no service techs (but that may be an East coast thing.) We've been happy with the EDAX service, and I enjoyed their user school very much. By the way, our SEM is a variable pressure JEOL 5900, and it's integrated with the EDAX system.
Hope I've been helpful,
Jane L. LaGoy Development Engineer Bodycote IMT, Inc. 155 River Street Andover, MA 01810 978-470-1620 jlagoy-at-bodycote-imt.com
Hi All: I have been asked to examine some films on a Si carbide substrate using TEM. Does anyone out there work with this material. Can one mechanically thin it using diamond films? Please offer some suggestions of what you are doing. Thanks in advance, Michael Coviello University of Texas Arlington
We have some staining that suggests the staining dye has transferred from one place to another and I write to see if anyone could shed some light on this for us.
In the experiment we stain isolated chromosomes with DAPI, rinse them, and then introduce them into plant protoplast cells (likely one or few per cell). There is no autofluroescence in the DAPI channel and we can follow the course of the dyed chromosomes over time. At first the DAPI fluorescence appears in the cytosol on structures likely to be the introduced chromosomes. Then it appears in the nucleus, where all the chromosomes are stained! This is especially evident when we culture these cells. Dividing cells at metaphase show DAPI fluorescence over the entire metaphase plate. Note that only some cells show DAPI fluorescence, consistent with the presumption that our fusion process that introduces the chromosomes is only successful in some of the cells.
Has anyone had an experience where dye has been transferred from one structure to another in a living cell? What are some alternative interpretations of this phenomenon?
Thanks for your comments,
Howard Berg
R. Howard Berg, Ph.D. Director, Integrated Microscopy Facility, Associate Member Donald Danforth Plant Science Center/Nidus Center 893 North Warson St. Louis, MO 63141
We tripod polished thin films on SiC. It works OK with diamond lapping film and diamond powder slurry as a final polish, but it takes a loooonnng time. Put a sacrificial piece of SiC on the second side tool pedestal so the final wedge polish will go from SiC sample to SiC tool. Otherwise the glass pedestal polishes faster than the SiC and makes a little step that can lead to sample breakage. ...Always a good idea to match the pedestal hardness to the specimen--hard or soft.
As I recall, we had more problems with poor film adhesion to the SiC than with polishing the SiC substrate.
Although we haven't tried it, I suppose a FIB would work fine.
Ron
There was a thread recently on scanners for TEM film. I have looked up all the models mentioned, on the web and called agents for prices - and produced a comparative table, given below.
I do not guarantee that the figures are accurate but they are my best interpretation of the data given.
In the light of experience and Nestor's comments, I would suggest that 2000 dpi is a minimum for TEM negatives. You may be able to get away with less nine times out of ten, but there will be occasions when you need more. I would exclude the Minolta and all the Epsons from consideration (despite the incredibly low prices of some of the Epsons) because of the low pixel density.
Among the rest the Nikon has the best pixel density and the best optical density (another critical parameter for TEM negatives). The price is very competitive too. The Nikon web site does not give a time for scanning a negative. On the face of it the Nikon would be a best buy - get a separate, inexpensive flatbed scanner for the other work.
These comments are all my own opinions based on manufacturers' data. Since we are considering purchase any comments to the contrary would be most welcome.
Code Maker Model Type
A Agfa DuoScan T2500 Flatbed -Transparency included
B Epson 1640 several versions Flatbed -Transparency option 1680 several versions
C 1600 several versions Flatbed -Transparency included
D Imacon Flextight Precision II Drum -for film and large format
E Minolta Dimage ScanMulti II Film
F Nikon Super Coolscan 8000ED Film
G Polaroid 45 Ultra Film
H Umax Powerlook 3000 Flatbed -Transparency included
Code dpi OD Time Price Opinion at 6 x 9 cm
A 2500 x2500 3.4 3 min $4,500 Fair
B 1600 x 3200 3.6 $300-$3000 Poor $800-$1400 Poor
C 1600 x 3200 3.3 $650-$1160 Not suitable
D 2240 x2240** 3.9/4.1 N/A above $10k Good: low pixel density
E 1128 x 1128 3.6 Not suitable
F 4000 x 4000 4.2 N/A $2,695 V. Good
G 2500 x 2500 3.8 5 min $7,495 Good but pricey
H 3048 x 3048 3.6 3 min $6,499 Good
-- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
I too am about to buy and I would make a couple of comments on your evaluation. First, let me remind everyone that the Dynamic range is a log scale so small numerical differences are significant.
I also think the Nikon Coolscan 8000 looks great but it only takes a 2.5 x 3.5 negative which is smaller than my JEOL and Hitachi EM negative sizes (~ 3 1/2 by 4 1/2"). Have these EM manufacturers gone to a smaller film size or is Nikon using a non-Japanese EM as their standard? seems odd but I don't see how the Nikon would be very useful. You say a {2000 line scanner would be useful 9 out of 10 times but want the 2000+ lines for the occasional high res scan. I would argue that the size of the negative was the more important variable to be worried about. The Nikon couldn't handle 4x5 LM negatives or transparencies from autoradiography of Westerns/Northerns, etc.
My leading candidate is the ArtixScan 1100 has a Dmax of 3.9 (about $1600 with SCSI card). This was has a 1000 x 2000 dpi resolution. more details at www.microtek.com. This is my leading candidate. It was 4 negative carriers and I await word whether one could be modified to carry a 3 1/2 by 4 1/2 negative. At worst, I will have my scientific instrumentation shop guys fabricate a holder. It comes with a glass 8 x 10 glass carrier for odd size negs but I want to avoid Newton rings and want a glassless carrier.
I would appreciate comments on the following argument (I think I have this correctly figured out but am not sure since so many out there seem to want to have a higher resolution scanner). I have a Fuji Pictrography 3000 printer with a 400 dpi output that is as good as any other widely available printer in the academic world. If you figure the maximum published image size is about 8 inches, that would mean the maximum image size be 3200 dpi wide. A 1000 dpi scan of my negative would be 4500 x 3500 dpi. I could crop by about 28% or 10% depending on the orientation of the negative and still be taking full advantage of the printer resolution. In reality, most EM publication prints are smaller than 8" wide so one could crop even more and still not need more than 1000 dpi. A resolution } 1000 dpi would be useful for subtle morphometric analysis but a 4000 dpi scan of a 3 x 4 negative would be 192 MB. That is pretty big for doing morphometry on! A 1000 dpi scan of a 3.5 x 4.5" negative would be about 16 MB and that is much more manageable. Perhaps the difference is in the type of EM we are doing. I am working with biological specimens doing standard thin section type stuff. are you doing some Material Sci application that demands more?
I will be interested in Alwyn (and any others) reply since I hope to buy one soon!
} . } } } There was a thread recently on scanners for TEM film. I have looked up } all the models mentioned, on the web and called agents for prices - and } produced a comparative table, given below. } } I do not guarantee that the figures are accurate but they are my best } interpretation of the data given. } } In the light of experience and Nestor's comments, I would suggest that } 2000 dpi is a minimum for TEM negatives. You may be able to get away } with less nine times out of ten, but there will be occasions when you } need more. } I would exclude the Minolta and all the Epsons from consideration } (despite the incredibly low prices of some of the Epsons) because of the } low pixel density. } } Among the rest the Nikon has the best pixel density and the best optical } density (another critical parameter for TEM negatives). The price is } very competitive too. The Nikon web site does not give a time for } scanning a negative. On the face of it the Nikon would be a best buy - } get a separate, inexpensive flatbed scanner for the other work. } } These comments are all my own opinions based on manufacturers' data. } Since we are considering purchase any comments to the contrary would be } most welcome. } } } } Code Maker Model Type } } } } A Agfa DuoScan T2500 Flatbed } -Transparency included } } B Epson 1640 several versions Flatbed } -Transparency option } 1680 several versions } } C 1600 several versions Flatbed } -Transparency included } } D Imacon Flextight Precision II Drum -for } film and large format } } E Minolta Dimage ScanMulti II Film } } F Nikon Super Coolscan 8000ED Film } } G Polaroid 45 Ultra Film } } H Umax Powerlook 3000 Flatbed } -Transparency included } } } } } } Code dpi OD Time Price } Opinion } at 6 x 9 cm } } } A 2500 x2500 3.4 3 min } $4,500 Fair } } B 1600 x 3200 3.6 } $300-$3000 Poor } } $800-$1400 Poor } } C 1600 x 3200 3.3 } $650-$1160 Not suitable } } D 2240 x2240** 3.9/4.1 N/A above } $10k Good: low pixel density } } E 1128 x 1128 3.6 } Not suitable } } F 4000 x 4000 4.2 N/A } $2,695 V. Good } } G 2500 x 2500 3.8 5 min } $7,495 Good but pricey } } H 3048 x 3048 3.6 3 min } $6,499 Good } } } -- } .......... } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University } 5 East Packer Avenue } Bethlehem } Pennsylvania 18015-3195 } Phone 610 758 4231 } Fax 610 758 4244 } jae5-at-lehigh.edu
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I assume that your SiC is single crystal. The small angle cleavage technique works for SiC. See MRS Proceedings, TEM Prep IV Vol 480.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Michael Coviello [mailto:coviello-at-mae.uta.edu] } Sent: Tuesday, April 10, 2001 10:59 AM } To: listserver } Subject: TEM-SiC wafer sample prep? } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } Hi All: } I have been asked to examine some films on a Si carbide } substrate using } TEM. Does anyone out there work with this material. Can one } mechanically thin it using diamond films? Please offer some } suggestions } of what you are doing. } Thanks in advance, } Michael Coviello } University of Texas Arlington } } }
Microscopy Scientist for two Recently Established NIH Centers for Biomedical Research at The University of Wyoming
As part of our seven interrelated projects into the biology, chemistry, and molecular biology of cardiovascular function and nitric oxide we seek a skilled microscopist to be hired at the non-tenured Assistant Research Professor level. Experience and research interests in state-of-the-art microscopy, including confocal and epifluorescence, and ultrastructural techniques. Primary responsibility will be management of the University's Microscopy Center. Opportunity includes the possibility of establishing a research program within this Center. Appointment will be at a (non-tenure track) Assistant Professor level for a renewable four year term.
Job Description: The person we seek will be responsible for organization of a new research laboratory facility at the University of Wyoming which will include two confocal microscopes and an electron microscope. The position includes overall management of the microscope facility, user training, and user supervision. Requirements for the position include experience with light, confocal, and transmission electron microscopy. This individual will oversee all aspects of specimen accession and processing, operation of the microscopes, photography, and record keeping. The individual will work with only minimum supervision. Responsibilities include: Serve as the technical manager of the facility and be responsible for the operation and maintenance of the confocal and EM microscope facility. In addition the manager will perform preventative maintenance on the equipment; maintain the lab, order supplies, schedule instruments, and oversee billing. Image analysis at the light, confocal and electron microscopic levels and preparation of micrographs for publication. Applicant Qualifications: Experience focus on both confocal and EM. Regarding confocal microscopy, we require experience with confocal and digital imaging techniques, visualization of living cells containing fluorescent probes, photobleaching, and fluorescence in situ hybridization. The successful applicant will have experience with tissue preparation for EM and the maintenance of an electron microscope. Excellent interpersonal and organizational skills are essential. Ph.D.. degree required Desirable Experience: Expertise and training in the operation of confocal microscope and EM microscope systems is required. Familiarity with light microscopy methods, immunofluorescent staining, use of fluorescent probes for physiologic measurements and the general principles of cell biological research are desirable. Significant facility with computers is desired. Salary Range: Commensurate with experience.
For additional information see our websites: www.uwyo.edu/nocobre www.uwyo.edu/MolecBio/Cobre To apply send complete CV, three references, and a cover letter indicating which position(s) you are applying for to: Lynda Payne, Department of Chemistry, University of Wyoming, Laramie, WY 82071-3838, USA. The searches will remain open until all positions are filled.
The University of Wyoming is located in a high (2,200 m) valley surrounded by the Rocky Mountains in the southeast corner of Wyoming. The University of Wyoming is an equal opportunity/affirmative action employer.
The Beam, newsletter for the Southeastern Microscopy Society (SEMS) is now online at the SEMS website in PDF format: http://www.biotech.ufl.edu/sems/
It contains information on the upcoming meeting in Clemson, SC. If you are a member, and have not received this notice via e-mail, and wish to be informed about the society through e-mail, please respond off-listserve at: jshields-at-cb.uga.edu
John Shields Center for Ultrastructural Research Univ. of Georgia Athens, GA
Such an instrument could provide for low-angle rotary shadowing capability for visualizing purified proteins at high resolution. It would need to have an electron beam gun for platinum evaporation and a separate one for carbon coating.
If this is what you have in mind, I support it. I purchased such an instrument that had good performance for about $20,000 about 10 years ago.
Dr. Joseph C. Besharse Professor and Chairman Dept of Cell Biology, Neurobiology and Anatomy Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226-0509
} From: Gang Ning {gning-at-mcw.edu} } Organization: Medical College of Wisconsin } Reply-To: gning-at-mcw.edu } Date: Tue, 10 Apr 2001 14:17:42 -0500 } To: Microscopy Newsgroup {Microscopy-at-sparc5.microscopy.com} } Cc: "Dr. Traktman" {ptrakt-at-post.its.mcw.edu} , Ming Lei {mlei-at-mcw.edu} , "Dr. } Besharse" {jbeshars-at-mcw.edu} } Subject: Sputter coater } } Hi All: } } I want to buy a new/used sputter coater which enables to do rotary } shadowing as well as carbon coating. Any suggestions/input are } appreciated. } } Greg Ning } } EM Facility } Medical College of Wisconsin }
Question: Hi, I have just purchased a microscope, a good biological one. I need a microscope kit but nobody sells them around here. Anyway already I have glass slides and covers. I have read some on microscopy and I need an adhesive, resin I think its called to prepare slides? is this true? Also some ink to stain specimens. What are the names of all these chemicals so I can buy them all seperatly since no one sells them all together. Thankyou.
Dear Theresa: Although a metallographic laboratory, we use alcohol for specimen preparation, mainly for preparation of etchants and for specimen cleaning. We found the paperwork associated with pure ethanol onerous and switched to denatured alcohol Type 3A with no discernable difference. Beware of some of the denaturants, they produce unusual side effects.
Sam Purdy National Steel Tech Center Trenton MI
} ---------- } From: BOES,TERESA (HP-Corvallis,ex1) } Sent: Monday, April 9, 2001 2:50 PM } To: 'microscopy-at-MSA.microscopy.com' } Subject: Substitutes for absolute ethanol? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Has anyone used any of the denatured ethanols as a substitute for absolute } ethanol? } } We have recently run into some difficulty when needing to reorder 200 } proof } ethanol, which we use for dehydration and infiltration of samples } (primarily } many types of paper) prior to embedding in Spurrs epoxy, and for cleaning } samples (non paper) and lenses of light microscopes. The chemical company } selling the ethanol is insisting that we must have a liquor license before } they will ship to us. } } Ethanol denatured with a variety of substances is readily available and } can } be shipped with no licensing requirements. Our concern is that the } denaturing agent will leave a detectable residue on lenses, samples, and } may } cause problems with the polymerization of Spurrs. Rather than obtaining a } liquor license, we are considering using one of the 100:5 ethanol: } methanol } blends. If any of you have had successful or unsuccessful experiences } substituting denatured ethanol for absolute in embedding or cleaning } protocols, I would appreciate hearing from you. } } Teresa Boes } Hewlett-Packard } Analytical and Development Lab } 1000 Circle Blvd } Corvallis, OR 97330 } 541-715-7055 } teresa_boes-at-hp.com } }
I would caution biased opinions from installation engineer and sales representatives ie... James Fotinopoulos.... www.semguy.com...
Hmmmmmmmmm?
Food for thought
----- Original Message -----
} From: "jeanross" {jeanross-at-blue.weeg.uiowa.edu} } To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} } Sent: Tuesday, April 10, 2001 10:22 AM } Subject: EDS summary } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I have put together a summary of responses I got from my inquiry about a } week } } or so ago about EDS systems. I really appreciate everyone's input. We } } haven't made any decisions yet since we are still gathering information } but } } your responses will help. I've included the responses in their entirety } so I } } hope this helps others as well. } } } } Thanks again from everyone who contributed. } } } } Jean Ross } } Central Microscopy Research Facility } } University of Iowa } } } } -------------------------------------------------------------------------- } --- } } I have been using an IXRF EDS / Gresham Detector (with IXRF digital pulse } } processor) system for almost 2 years. Initially, the IXRF was installed on } } an ETEC. About 15 months ago the Etec was replaced with a Hitachi 3500, a } } new detector (Gresham) was purchased, and the EDS was installed on the } 3500. } } Generally, I am quite satisfied. There are minor software bugs, but IXRF } } has been reasonably good at fixing them when discovered. It has been my } } experience that all the systems have bugs, perhaps some more than others. } } Prior to the IXRF, I had a Kevex 8000/Delta. } } } } Low end noise and broad peaks were evident on first installation, but were } } soon fixed by tweaking the detector preamp and pulse processor amp. } } } } I am still running their first software package, "Iridium". I have the } } newest release, "EDS 2000", but lack of time has kept me from installing } and } } checking it out. } } } } I should mention that IXRF is a "virtual" company, with people spread out } } between Texas, California, etc. This has not proven to be a problem. } } } } Woody White } } McDermott Technology, Inc. } } nwwhite-at-mcdermott.com } } } } -------------------------------------------------------------------------- } --- } } } } } } We have had EDAX for about a decade and a half, and we are very pleased } with } } the product and the service we get. } } Carol Heckman } } heckman-at-bgnet.bgsu.edu } } } } -------------------------------------------------------------------------- } --- } } } } } } look at IXRF eds systems there web site is www.ixrfsystems.com, they are } } very affordable and offer no nonsense performance that second to none. } } } } happy ixrf user, } } } } James Fotinopoulos } } yzfrjim-at-ix.netcom.com } } } } } } -------------------------------------------------------------------------- } ---- } } } } I would recommend you give consideration to Doug Connors at } } TN Analyzer Service, Inc. of Dane, WI. Doug has rebuild and } } upgraded detectors for me for the last 6 years. He is dependable } } knowledgeable, and economical as well. } } } } Bob Roberts } } EM Lab Services, Inc. } } 2409 S. Rural Rd Suite C } } Tempe, Arizona 85282 } } (480) 967-3946 } } bobrobs-at-earthlink.net } } } } -------------------------------------------------------------------------- } --- } } } } } } We recently purchased a Noran Instruments Vantage DS1. We purchased this } } based on some very impressive demonstrations of software that the } salesperson } } brought in. Unfortunately they are still working out the bugs in their } } software. Everything they have is ported over from Unix, and literally } runs } } in a unix shell on an Microsoft Windows NT platform. This makes their } } software fairly buggy. Their response time to fix major bugs and hang-ups } in } } the software has been very slow, and if given the opportunity to do it all } } over again I'd probably look at Oxford Instruments. I would still rate } the } } quality of the equipment very high. Our detector performs at the } specified } } resolution, and is a good piece of equipment. Now if they could only get } the } } software end of it straight... } } My vote: } } 1) Oxford Instruments } } 2) Noran Instruments } } 3) Edax or some of the smaller players } } The benefit with going with a larger company is support and upgrades. We } have } } a 10 year old WDX that we just purchased new software and interface for } last } } year. Our old EDS was given some trade in value by Noran. And we all } know } } how valuable a service contract can be... } } } } Get back to me if you have any more questions, } } ~Jonathan } } Jonathan Dunlap } } Analytical Laboratory Manager } } Osram Sylvania Inc. } } 816 Lexington Avenue } } Warren, PA 16365 } } Ph: 814-726-6991 } } Fax: 814-726-6942 } } Jonathan.Dunlap-at-sylvania.com } } } } } } -------------------------------------------------------------------------- } ---- } } } } } } We have an Oxford Instruments Link ISIS Model 200 on our 2460N. We have } been } } happy with it, but I don't know about the direction that Oxford is } heading. I } } don't care for the feel of their new INCA software. Some might like it. It } } also seems to be slow coming together. Some of the functions are still } lacking } } after 2 (or is it 3) years of seeing it at MSA. } } } } We have an IXRF system on our JEOL 840A. It was a good price ($30K) for an } } upgrade to our Kevex several years ago. It does what we need. They keep at } } work on the software and have it freely available on the web. I might have } to } } pay closer attention and stay away from the beta stuff. They are still } working } } on it. They also have a nice digital pulse processor which stills stand } alone } } for about $5k. } } } } I still feel funny about some contacts with EVEX. I can't say much about } EDAX, } } NORAN, or PGT. They should all have good stuff but it might be pricey. The } } last we seriously looked at them was 6 years ago or so when we opted for } the } } Oxford. } } } } I was intrigued by the unit from Quartz PCI. I think it was called X-ray } One, } } or such. It was new at MSA 1-1/2 years ago but looked promising. } } } } Feel free to call if you want more details. } } } } Warren E Straszheim } } wesaia-at-iastate.edu } } } } -------------------------------------------------------------------------- } --- } } } } } } We purchased an EDAX Falcon system for our Hitachi S-3000N and I've been } } pleased with it. It has better light element sensitivity than most which } } was very important to me although I don't think that its mapping } } capabilities are as good as PGT's, say. I don't have direct experience } with } } Noran although I did talk to them and their system seemed ok - but } logistics } } didn't favor Noran so I passed on them. EDAX does have good integration } } with the Hitachi and the Quartz database. } } } } I'd be glad to respond more specifically if you'd like. } } } } Richard Shalvoy } } Arch Chemicals } } Cheshire, CT } } RBShalvoy-at-archchemicals.com } } } } -------------------------------------------------------------------------- } --- } } } } } } I have an iXRF systems out of Texas using a Gresham detector. It works } } well. Not the most cutting edge, but they are one of the "start ups". They } } have been around for I guess 6-7 years. I have a digital pulse processor } } and completely active control for x-ray maps and such. They are very price } } competitive, but lack a dedicated technical support person. You talk to } the } } programmer or electronics expert, but no techs on the phone whenever you } } have a software question. But, if you willing to wait a day for some } } answers then they are worth it. I haven't run across the problem where I } } thought, "if I just had a better system". If you want to integrated w/ WDS } } than maybe Noran. Also, if you want to integrated w/ motorized stage } } control, I don't think they off such a package, like the bigger companies. } } } } I have a Hitachi 450. I used to run a 2400 and 500 before I quit my day } job } } and went out on my own. I am very happy w/ Hitachi. } } } } Good Luck } } } } Fell free to call with any specifics. } } } } Their web page is www.ixrfsystems.com } } } } Ric } } } } SMARTech } } 860-491-3299 } } www.semguy.com } } 19 Cornwall Drive } } Goshen CT 06756 } } smartech-at-javanet.com } } } } -------------------------------------------------------------------------- } ---- } } } } } } Hello, all: } } } } I use Oxford ISIS300 system on HITACHI S-3500N (with VP mode) for light } } element analysis, mostly C, O, N, F, P, S, Si, Mg, as well as metal Co, } Ni-P, } } Pt, Cr, Fe, W, etc. This system works well. One useful function is the } overlay } } of 2 spectrums. I can easily subtract the blank from the sample spot and } make } } it easy to identify what is (are) in the sample. I am sure some other } program } } may have this kind of function, but I have not seen. } } } } Zhiyu Wang } } zhiyuw-at-home.com } } I would be interested in seeing the responses as I am going to try and get } } funding next year for a replacement for our EDAX PV9100 on an Hitachi } s-450. } } } } Dave } } David.Patton-at-uwe.ac.uk } } } } -------------------------------------------------------------------------- } --- } } } } } } We have been running EDX on SEMs and TEMs for many years. We used to have } a } } range of systems from Kevex, PGT, Noran, Link, EDAX, however a few years } ago } } we decided that we ought to standardize on one common system. After } evaluation } } we bought three Oxford Instruments ISIS systems. Whenever we have upgraded } or } } bought new systems they have been Oxford Instruments ISIS or now INCA. } } } } I have been happy with the ISIS except for the file handling that was not } } designed for a multi user facility such as ours (approx 120 EM users in } total } } roughly 25 to 30 swapping every year). I am really quite impressed by the } } INCA, Oxford Instruments are, at last, listening to the users and adding } user } } requested facilities. They have sorted out the file handling mess of the } ISIS } } and structured it well for an SEM user (not quite as well for a TEM user } but } } there are less of us). The software structure is quite intuitive and there } is } } a really impressive help menu and explanation of everything from the } physics } } of X-ray generation, how EM's work, how detectors work and how to analyze } } samples. } } } } Their detectors have always been good and the SATW (thin window detectors) } } still have a reasonable efficiency at low Z. B is possible but C is easy } and } } even the N peak is over 30% efficient (there is often a high absorption at } N). } } } } Another feature that is invaluable for TEM is the integral shutter that } will } } close when the count rate is too high. This protects the crystal, it } prevents } } it overloading and shutting down or worse the crystal efficiency may } change } } for a few minutes until it recovers fully. This may affects your } quantitative } } work. In TEM this is usually caused by hitting the grid bar and not really } a } } problem in SEM but I don't know what secondaries and ions you will have in } a } } variable pressure SEM. It could be useful for you, check with other high } } pressure SEM users. } } } } Regards, } } Ron } } } } Please note: Oxford Instruments have upgraded an ISIS to an INCA system in } my } } department, without charge, in return for access to the instrument for } } development projects and demonstrations for a fixed number of days. I } receive } } no benefit from this and the department has no benefit from Oxford } Instruments } } sales. I remain a thorn in the flesh of all our suppliers if I think they } } could improve their products or service. } } ron.doole-at-materials.oxford.ac.uk } } } } } } -------------------------------------------------------------------------- } ---- } } } } } } Hello, } } I am very familiar with the Oxford ISIS 300 series spectrometers. They are } } ok, and the new Inca system looks good too. However, I recently saw the } PGT } } spectrometer at Lehigh and it is very impressive. } } } } Steve } } Stephen_Skirius-at-bkitech.com } } } } } } } } -------------------------------------------------------------------------- } } } } } } I'm also in the market for an EDS system and have looked at EDAX, Noran, } } PGT and Oxford. } } } } I edited out PGT because in order to quantify you have to optimize the } } system for the type of sample by playing with fudge factors, which none of } } the other systems have to do (though one of them, I think Noran, lets you } } adjust a sensitivity factor if you want to, but they didn't do it on my } } samples that were tested against known microprobe results and the answers } } were fine). I also eliminated Oxford, though it has a terrific user } } interface (maybe at the expense of functionality), because they } } consistently IDed my aluminum peaks as Br or Tm (!); this made me wonder } } about all their algorithms. They claim it had to do with the takeoff angle } } on the particular SEM being used, but that shouldn't be a factor. } } } } I like both EDAX and Noran, though for different reasons. EDAX user } } interface is better than Noran's, though again, I think Noran possibly } } offers more routines (it's hard to keep track and see absolutely } everything } } a system has to offer in a demo day....).Noran can multitask - work on } } several programs while a spectral map is being collected, for instance } } (does EDAX? I have to check). But EDAX has a beam skirt reduction routine } } for low vac mode (though it's time consuming, so a bit cumbersome), and } } their peak modeling is right up front - but Noran can put theirs up front } } also if you want to have it accessible (yes) and I think Noran might be a } } little better engineered. } } } } As you can see I'm still in a quandary (ditto for the two contending SEMs, } } LEO and ESEM). Whatever I decide I'll still be very interested in the } } results of your posting - especially if other folks' info comes in within } } the next week or so it would help in my decision too. } } } } I hope my input helps a little. Good luck with your quest! } } } } Dee Breger } } micro-at-ldeo.columbia.edu } } } } } } -------------------------------------------------------------------------- } - } } } } } } I looked into Noran, EDAX, and PGT. Noran was quickly culled (less user } } friendly, less abilities, didn't work right during demo), but EDAX and PGT } } both seemed to have equivalent capabilities (PGT claimed a 'proprietary' } } signal amplifier/digitizer doohickey, but it was only in the placement.) } } For the long-time spectrum gathering (I forget the technical term), PGT } } makes many passes with short dwell times while EDAX dwells on each pixel } } much longer to collect data & does it in 1 pass. Kinda 6 of one, half } dozen } } of the other. What made us choose the EDAX Phoenix system was the fact } that } } the PGT software was UNIX-based (although hidden) while EDAX is PC-based. } } I've heard rumors that PGT is switching to PC-based; we purchased our } system } } in 1999. I've also heard that Noran has practically no service techs (but } } that may be an East coast thing.) We've been happy with the EDAX service, } } and I enjoyed their user school very much. By the way, our SEM is a } } variable pressure JEOL 5900, and it's integrated with the EDAX system. } } } } Hope I've been helpful, } } } } Jane L. LaGoy } } Development Engineer } } Bodycote IMT, Inc. } } 155 River Street } } Andover, MA 01810 } } 978-470-1620 } } jlagoy-at-bodycote-imt.com } } } } } } } } } } }
{HTML} {FONT FACE=arial,helvetica} Jean, {BR} {BR} I would caution biased opinions from installation engineer and sales {BR} representatives ie... James Fotinopoulos.... www.semguy.com... {BR} {BR} Hmmmmmmmmm? {BR} {BR} Food for thought {BR} {FONT SIZE=2} {BR} {BR} {BR} {BR} {BR} ----- Original Message ----- {BR} {/FONT} {FONT COLOR="#000000" SIZE=3 FAMILY="SANSSERIF" FACE="Arial" LANG="0"} {BR} {/FONT} {FONT COLOR="#000000" SIZE=2 FAMILY="SANSSERIF" FACE="Arial" LANG="0"} {BLOCKQUOTE TYPE=CITE style="BORDER-LEFT: #0000ff 2px solid; MARGIN-LEFT: 5px; MARGIN-RIGHT: 0px; PADDING-LEFT: 5px"} From: "jeanross" <jeanross-at-blue.weeg.uiowa.edu> {BR} To: "Microscopy Listserver" <Microscopy-at-sparc5.microscopy.com> {BR} Sent: Tuesday, April 10, 2001 10:22 AM {BR} Subject: EDS summary {BR} {BR} {BR} > ------------------------------------------------------------------------ {BR} > The Microscopy ListServer -- Sponsor: The Microscopy Society of America {BR} > To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {BR} > On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {BR} > -----------------------------------------------------------------------. {BR} > {BR} > {BR} > I have put together a summary of responses I got from my inquiry about a {BR} week {BR} > or so ago about EDS systems. I really appreciate everyone's input. We {BR} > haven't made any decisions yet since we are still gathering information {BR} but {BR} > your responses will help. I've included the responses in their entirety {BR} so I {BR} > hope this helps others as well. {BR} > {BR} > Thanks again from everyone who contributed. {BR} > {BR} > Jean Ross {BR} > Central Microscopy Research Facility {BR} > University of Iowa {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > I have been using an IXRF EDS / Gresham Detector (with IXRF digital pulse {BR} > processor) system for almost 2 years. Initially, the IXRF was installed on {BR} > an ETEC. About 15 months ago the Etec was replaced with a Hitachi 3500, a {BR} > new detector (Gresham) was purchased, and the EDS was installed on the {BR} 3500. {BR} > Generally, I am quite satisfied. There are minor software bugs, but IXRF {BR} > has been reasonably good at fixing them when discovered. It has been my {BR} > experience that all the systems have bugs, perhaps some more than others. {BR} > Prior to the IXRF, I had a Kevex 8000/Delta. {BR} > {BR} > Low end noise and broad peaks were evident on first installation, but were {BR} > soon fixed by tweaking the detector preamp and pulse processor amp. {BR} > {BR} > I am still running their first software package, "Iridium". I have the {BR} > newest release, "EDS 2000", but lack of time has kept me from installing {BR} and {BR} > checking it out. {BR} > {BR} > I should mention that IXRF is a "virtual" company, with people spread out {BR} > between Texas, California, etc. This has not proven to be a problem. {BR} > {BR} > Woody White {BR} > McDermott Technology, Inc. {BR} > nwwhite-at-mcdermott.com {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > {BR} > {BR} > We have had EDAX for about a decade and a half, and we are very pleased {BR} with {BR} > the product and the service we get. {BR} > Carol Heckman {BR} > heckman-at-bgnet.bgsu.edu {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > {BR} > {BR} > look at IXRF eds systems there web site is www.ixrfsystems.com, they are {BR} > very affordable and offer no nonsense performance that second to none. {BR} > {BR} > happy ixrf user, {BR} > {BR} > James Fotinopoulos {BR} > yzfrjim-at-ix.netcom.com {BR} > {BR} > {BR} > -------------------------------------------------------------------------- {BR} ---- {BR} > {BR} > I would recommend you give consideration to Doug Connors at {BR} > TN Analyzer Service, Inc. of Dane, WI. Doug has rebuild and {BR} > upgraded detectors for me for the last 6 years. He is dependable {BR} > knowledgeable, and economical as well. {BR} > {BR} > Bob Roberts {BR} > EM Lab Services, Inc. {BR} > 2409 S. Rural Rd Suite C {BR} > Tempe, Arizona 85282 {BR} > (480) 967-3946 {BR} > bobrobs-at-earthlink.net {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > {BR} > {BR} > We recently purchased a Noran Instruments Vantage DS1. We purchased this {BR} > based on some very impressive demonstrations of software that the {BR} salesperson {BR} > brought in. Unfortunately they are still working out the bugs in their {BR} > software. Everything they have is ported over from Unix, and literally {BR} runs {BR} > in a unix shell on an Microsoft Windows NT platform. This makes their {BR} > software fairly buggy. Their response time to fix major bugs and hang-ups {BR} in {BR} > the software has been very slow, and if given the opportunity to do it all {BR} > over again I'd probably look at Oxford Instruments. I would still rate {BR} the {BR} > quality of the equipment very high. Our detector performs at the {BR} specified {BR} > resolution, and is a good piece of equipment. Now if they could only get {BR} the {BR} > software end of it straight... {BR} > My vote: {BR} > 1) Oxford Instruments {BR} > 2) Noran Instruments {BR} > 3) Edax or some of the smaller players {BR} > The benefit with going with a larger company is support and upgrades. We {BR} have {BR} > a 10 year old WDX that we just purchased new software and interface for {BR} last {BR} > year. Our old EDS was given some trade in value by Noran. And we all {BR} know {BR} > how valuable a service contract can be... {BR} > {BR} > Get back to me if you have any more questions, {BR} > ~Jonathan {BR} > Jonathan Dunlap {BR} > Analytical Laboratory Manager {BR} > Osram Sylvania Inc. {BR} > 816 Lexington Avenue {BR} > Warren, PA 16365 {BR} > Ph: 814-726-6991 {BR} > Fax: 814-726-6942 {BR} > Jonathan.Dunlap-at-sylvania.com {BR} > {BR} > {BR} > -------------------------------------------------------------------------- {BR} ---- {BR} > {BR} > {BR} > We have an Oxford Instruments Link ISIS Model 200 on our 2460N. We have {BR} been {BR} > happy with it, but I don't know about the direction that Oxford is {BR} heading. I {BR} > don't care for the feel of their new INCA software. Some might like it. It {BR} > also seems to be slow coming together. Some of the functions are still {BR} lacking {BR} > after 2 (or is it 3) years of seeing it at MSA. {BR} > {BR} > We have an IXRF system on our JEOL 840A. It was a good price ($30K) for an {BR} > upgrade to our Kevex several years ago. It does what we need. They keep at {BR} > work on the software and have it freely available on the web. I might have {BR} to {BR} > pay closer attention and stay away from the beta stuff. They are still {BR} working {BR} > on it. They also have a nice digital pulse processor which stills stand {BR} alone {BR} > for about $5k. {BR} > {BR} > I still feel funny about some contacts with EVEX. I can't say much about {BR} EDAX, {BR} > NORAN, or PGT. They should all have good stuff but it might be pricey. The {BR} > last we seriously looked at them was 6 years ago or so when we opted for {BR} the {BR} > Oxford. {BR} > {BR} > I was intrigued by the unit from Quartz PCI. I think it was called X-ray {BR} One, {BR} > or such. It was new at MSA 1-1/2 years ago but looked promising. {BR} > {BR} > Feel free to call if you want more details. {BR} > {BR} > Warren E Straszheim {BR} > wesaia-at-iastate.edu {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > {BR} > {BR} > We purchased an EDAX Falcon system for our Hitachi S-3000N and I've been {BR} > pleased with it. It has better light element sensitivity than most which {BR} > was very important to me although I don't think that its mapping {BR} > capabilities are as good as PGT's, say. I don't have direct experience {BR} with {BR} > Noran although I did talk to them and their system seemed ok - but {BR} logistics {BR} > didn't favor Noran so I passed on them. EDAX does have good integration {BR} > with the Hitachi and the Quartz database. {BR} > {BR} > I'd be glad to respond more specifically if you'd like. {BR} > {BR} > Richard Shalvoy {BR} > Arch Chemicals {BR} > Cheshire, CT {BR} > RBShalvoy-at-archchemicals.com {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > {BR} > {BR} > I have an iXRF systems out of Texas using a Gresham detector. It works {BR} > well. Not the most cutting edge, but they are one of the "start ups". They {BR} > have been around for I guess 6-7 years. I have a digital pulse processor {BR} > and completely active control for x-ray maps and such. They are very price {BR} > competitive, but lack a dedicated technical support person. You talk to {BR} the {BR} > programmer or electronics expert, but no techs on the phone whenever you {BR} > have a software question. But, if you willing to wait a day for some {BR} > answers then they are worth it. I haven't run across the problem where I {BR} > thought, "if I just had a better system". If you want to integrated w/ WDS {BR} > than maybe Noran. Also, if you want to integrated w/ motorized stage {BR} > control, I don't think they off such a package, like the bigger companies. {BR} > {BR} > I have a Hitachi 450. I used to run a 2400 and 500 before I quit my day {BR} job {BR} > and went out on my own. I am very happy w/ Hitachi. {BR} > {BR} > Good Luck {BR} > {BR} > Fell free to call with any specifics. {BR} > {BR} > Their web page is www.ixrfsystems.com {BR} > {BR} > Ric {BR} > {BR} > SMARTech {BR} > 860-491-3299 {BR} > www.semguy.com {BR} > 19 Cornwall Drive {BR} > Goshen CT 06756 {BR} > smartech-at-javanet.com {BR} > {BR} > -------------------------------------------------------------------------- {BR} ---- {BR} > {BR} > {BR} > Hello, all: {BR} > {BR} > I use Oxford ISIS300 system on HITACHI S-3500N (with VP mode) for light {BR} > element analysis, mostly C, O, N, F, P, S, Si, Mg, as well as metal Co, {BR} Ni-P, {BR} > Pt, Cr, Fe, W, etc. This system works well. One useful function is the {BR} overlay {BR} > of 2 spectrums. I can easily subtract the blank from the sample spot and {BR} make {BR} > it easy to identify what is (are) in the sample. I am sure some other {BR} program {BR} > may have this kind of function, but I have not seen. {BR} > {BR} > Zhiyu Wang {BR} > zhiyuw-at-home.com {BR} > I would be interested in seeing the responses as I am going to try and get {BR} > funding next year for a replacement for our EDAX PV9100 on an Hitachi {BR} s-450. {BR} > {BR} > Dave {BR} > David.Patton-at-uwe.ac.uk {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > {BR} > {BR} > We have been running EDX on SEMs and TEMs for many years. We used to have {BR} a {BR} > range of systems from Kevex, PGT, Noran, Link, EDAX, however a few years {BR} ago {BR} > we decided that we ought to standardize on one common system. After {BR} evaluation {BR} > we bought three Oxford Instruments ISIS systems. Whenever we have upgraded {BR} or {BR} > bought new systems they have been Oxford Instruments ISIS or now INCA. {BR} > {BR} > I have been happy with the ISIS except for the file handling that was not {BR} > designed for a multi user facility such as ours (approx 120 EM users in {BR} total {BR} > roughly 25 to 30 swapping every year). I am really quite impressed by the {BR} > INCA, Oxford Instruments are, at last, listening to the users and adding {BR} user {BR} > requested facilities. They have sorted out the file handling mess of the {BR} ISIS {BR} > and structured it well for an SEM user (not quite as well for a TEM user {BR} but {BR} > there are less of us). The software structure is quite intuitive and there {BR} is {BR} > a really impressive help menu and explanation of everything from the {BR} physics {BR} > of X-ray generation, how EM's work, how detectors work and how to analyze {BR} > samples. {BR} > {BR} > Their detectors have always been good and the SATW (thin window detectors) {BR} > still have a reasonable efficiency at low Z. B is possible but C is easy {BR} and {BR} > even the N peak is over 30% efficient (there is often a high absorption at {BR} N). {BR} > {BR} > Another feature that is invaluable for TEM is the integral shutter that {BR} will {BR} > close when the count rate is too high. This protects the crystal, it {BR} prevents {BR} > it overloading and shutting down or worse the crystal efficiency may {BR} change {BR} > for a few minutes until it recovers fully. This may affects your {BR} quantitative {BR} > work. In TEM this is usually caused by hitting the grid bar and not really {BR} a {BR} > problem in SEM but I don't know what secondaries and ions you will have in {BR} a {BR} > variable pressure SEM. It could be useful for you, check with other high {BR} > pressure SEM users. {BR} > {BR} > Regards, {BR} > Ron {BR} > {BR} > Please note: Oxford Instruments have upgraded an ISIS to an INCA system in {BR} my {BR} > department, without charge, in return for access to the instrument for {BR} > development projects and demonstrations for a fixed number of days. I {BR} receive {BR} > no benefit from this and the department has no benefit from Oxford {BR} Instruments {BR} > sales. I remain a thorn in the flesh of all our suppliers if I think they {BR} > could improve their products or service. {BR} > ron.doole-at-materials.oxford.ac.uk {BR} > {BR} > {BR} > -------------------------------------------------------------------------- {BR} ---- {BR} > {BR} > {BR} > Hello, {BR} > I am very familiar with the Oxford ISIS 300 series spectrometers. They are {BR} > ok, and the new Inca system looks good too. However, I recently saw the {BR} PGT {BR} > spectrometer at Lehigh and it is very impressive. {BR} > {BR} > Steve {BR} > Stephen_Skirius-at-bkitech.com {BR} > {BR} > {BR} > {BR} > -------------------------------------------------------------------------- {BR} > {BR} > {BR} > I'm also in the market for an EDS system and have looked at EDAX, Noran, {BR} > PGT and Oxford. {BR} > {BR} > I edited out PGT because in order to quantify you have to optimize the {BR} > system for the type of sample by playing with fudge factors, which none of {BR} > the other systems have to do (though one of them, I think Noran, lets you {BR} > adjust a sensitivity factor if you want to, but they didn't do it on my {BR} > samples that were tested against known microprobe results and the answers {BR} > were fine). I also eliminated Oxford, though it has a terrific user {BR} > interface (maybe at the expense of functionality), because they {BR} > consistently IDed my aluminum peaks as Br or Tm (!); this made me wonder {BR} > about all their algorithms. They claim it had to do with the takeoff angle {BR} > on the particular SEM being used, but that shouldn't be a factor. {BR} > {BR} > I like both EDAX and Noran, though for different reasons. EDAX user {BR} > interface is better than Noran's, though again, I think Noran possibly {BR} > offers more routines (it's hard to keep track and see absolutely {BR} everything {BR} > a system has to offer in a demo day....).Noran can multitask - work on {BR} > several programs while a spectral map is being collected, for instance {BR} > (does EDAX? I have to check). But EDAX has a beam skirt reduction routine {BR} > for low vac mode (though it's time consuming, so a bit cumbersome), and {BR} > their peak modeling is right up front - but Noran can put theirs up front {BR} > also if you want to have it accessible (yes) and I think Noran might be a {BR} > little better engineered. {BR} > {BR} > As you can see I'm still in a quandary (ditto for the two contending SEMs, {BR} > LEO and ESEM). Whatever I decide I'll still be very interested in the {BR} > results of your posting - especially if other folks' info comes in within {BR} > the next week or so it would help in my decision too. {BR} > {BR} > I hope my input helps a little. Good luck with your quest! {BR} > {BR} > Dee Breger {BR} > micro-at-ldeo.columbia.edu {BR} > {BR} > {BR} > -------------------------------------------------------------------------- {BR} - {BR} > {BR} > {BR} > I looked into Noran, EDAX, and PGT. Noran was quickly culled (less user {BR} > friendly, less abilities, didn't work right during demo), but EDAX and PGT {BR} > both seemed to have equivalent capabilities (PGT claimed a 'proprietary' {BR} > signal amplifier/digitizer doohickey, but it was only in the placement.) {BR} > For the long-time spectrum gathering (I forget the technical term), PGT {BR} > makes many passes with short dwell times while EDAX dwells on each pixel {BR} > much longer to collect data & does it in 1 pass. Kinda 6 of one, half {BR} dozen {BR} > of the other. What made us choose the EDAX Phoenix system was the fact {BR} that {BR} > the PGT software was UNIX-based (although hidden) while EDAX is PC-based. {BR} > I've heard rumors that PGT is switching to PC-based; we purchased our {BR} system {BR} > in 1999. I've also heard that Noran has practically no service techs (but {BR} > that may be an East coast thing.) We've been happy with the EDAX service, {BR} > and I enjoyed their user school very much. By the way, our SEM is a {BR} > variable pressure JEOL 5900, and it's integrated with the EDAX system. {BR} > {BR} > Hope I've been helpful, {BR} > {BR} > Jane L. LaGoy {BR} > Development Engineer {BR} > Bodycote IMT, Inc. {BR} > 155 River Street {BR} > Andover, MA 01810 {BR} > 978-470-1620 {BR} > jlagoy-at-bodycote-imt.com {BR} > {BR} > {BR} > {BR} > {BR} {BR} {/XMP} {/FONT} {FONT COLOR="#0f0f0f" SIZE=2 FAMILY="SANSSERIF" FACE="Arial" LANG="0"} {BR} {/FONT} {/HTML}
Return-Path: {ptarq-at-evex.com} Received: from rly-xb03.mx.aol.com (rly-xb03.mail.aol.com [172.20.105.104]) by air-xb02.mail.aol.com (v77_r1.36) with ESMTP; Tue, 10 Apr 2001 17:56:34 -0500 Received: from mxr01.nyc01.dsl.net (mxr01.nyc01.dsl.net [216.175.203.53]) by rly-xb03.mx.aol.com (v77_r1.36) with ESMTP; Tue, 10 Apr 2001 17:56:05 -0400 Received: from pb133nt (64-51-105-68.client.dsl.net [64.51.105.68]) by mxr01.nyc01.dsl.net (Postfix) with SMTP id 1F49C34538 for {lhconsulting-at-aol.com} ; Tue, 10 Apr 2001 17:52:57 -0400 (EDT) Message-ID: {002b01c0c208$95dfe860$6601a8c0-at-evex.com} Reply-To: "Evex Mail" {ptarq-at-evex.com} } From: "Evex Mail" {ptarq-at-evex.com} To: {lhconsulting-at-aol.com}
I guess it was a slip: In a sputter coater you can do rotary coating but shadowing is possible in evaporators only. For carbon coating the sputter coater would need an attachment for carbon string evaporation, which may be used for SEM and analyses, but not in TEM. Disclaimer: ProSciTech is the EMITECH instrument distributor in Australasia. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Wednesday, April 11, 2001 5:18 AM, Gang Ning [SMTP:gning-at-mcw.edu] wrote: } } } Hi All: } } I want to buy a new/used sputter coater which enables to do rotary } shadowing as well as carbon coating. Any suggestions/input are } appreciated. } } Greg Ning } } EM Facility } Medical College of Wisconsin }
A colleague and I each recently bought Microtek scanners to scan TEM negatives. I have the Artixscan 1100 and he has the Model 8700 which has similar characteristics (actually higher resolution -1200dpi), 3.9 dmax at 42 bits color (14 grayscale), and the glassless film carrier setup. The 8700 has USB and Firewire interfaces and is cheaper ( {$1000), and the 1000 dpi Model 1100 has a SCSI interface. You might want to check out the specs of the lower cost model 8700 on the microtekusa website if your computer can handle USB or Firewire. Both scanners have performed up to our expectations, which I would characterize as modest. Microtek does not supply a 3-1/4 x 4 " negative carrier for standard size TEM film but you can easily make a serviceable one from stiff paper or light cardboard.
How much scanner resolution should you buy? The answer depends on how you intend to use it. Most applications do not require capturing the full resolution of the negative. From a practical viewpoint, the scanner resolution just determines how many times you can magnify the negative image to produce the final print size. For example, to get a publication-size print at 300 dpi, an image scanned at 1200 dpi scan could be zoomed 4X. A practical alternative to spending more for higher scanning resolution is to take photos at higher magnification. One exception is with lattice images from the TEM, which (depending on the lattice fringe spacing on the negative) might require higher scan resolutions to avoid getting a moire effect. (Of course, not everyone agrees. My colleague prefers to always scan at the maximum resolution).
What does a Dmax of 3.9 mean to you? To me it means a very dark negative. D is the log of the transmitted to incident intensity ratio. I wonder if users ever actually verify the manufacturer's specs with a calibrated density target. A Dmax of 3.9 can be useful for scanning TEM diffraction patterns that might have high contrast, but TEM micrograph negatives of metals and ceramics generally don't have that much contrast and biological thin section photos tend to have rather weak contrast. If your negatives are simply dark, use shorter photo exposure times. Scanning with maximum allowed grayscale resolutions (e.g., 14 bits rather than 8) is highly recommended if you intend to enhance or adjust images, but that's another story.
Larry Thomas Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto:Larry.Thomas-at-pnl.gov
---------- From: Tom Phillips Sent: Tuesday, April 10, 2001 11:22 AM To: Alwyn Eades Cc: Microscopy-at-sparc5.microscopy.com Subject: Re: Scanners
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I too am about to buy and I would make a couple of comments on your evaluation. First, let me remind everyone that the Dynamic range is a log scale so small numerical differences are significant.
I also think the Nikon Coolscan 8000 looks great but it only takes a 2.5 x 3.5 negative which is smaller than my JEOL and Hitachi EM negative sizes (~ 3 1/2 by 4 1/2"). Have these EM manufacturers gone to a smaller film size or is Nikon using a non-Japanese EM as their standard? seems odd but I don't see how the Nikon would be very useful. You say a {2000 line scanner would be useful 9 out of 10 times but want the 2000+ lines for the occasional high res scan. I would argue that the size of the negative was the more important variable to be worried about. The Nikon couldn't handle 4x5 LM negatives or transparencies from autoradiography of Westerns/Northerns, etc.
My leading candidate is the ArtixScan 1100 has a Dmax of 3.9 (about $1600 with SCSI card). This was has a 1000 x 2000 dpi resolution. more details at www.microtek.com. This is my leading candidate. It was 4 negative carriers and I await word whether one could be modified to carry a 3 1/2 by 4 1/2 negative. At worst, I will have my scientific instrumentation shop guys fabricate a holder. It comes with a glass 8 x 10 glass carrier for odd size negs but I want to avoid Newton rings and want a glassless carrier.
I would appreciate comments on the following argument (I think I have this correctly figured out but am not sure since so many out there seem to want to have a higher resolution scanner). I have a Fuji Pictrography 3000 printer with a 400 dpi output that is as good as any other widely available printer in the academic world. If you figure the maximum published image size is about 8 inches, that would mean the maximum image size be 3200 dpi wide. A 1000 dpi scan of my negative would be 4500 x 3500 dpi. I could crop by about 28% or 10% depending on the orientation of the negative and still be taking full advantage of the printer resolution. In reality, most EM publication prints are smaller than 8" wide so one could crop even more and still not need more than 1000 dpi. A resolution } 1000 dpi would be useful for subtle morphometric analysis but a 4000 dpi scan of a 3 x 4 negative would be 192 MB. That is pretty big for doing morphometry on! A 1000 dpi scan of a 3.5 x 4.5" negative would be about 16 MB and that is much more manageable. Perhaps the difference is in the type of EM we are doing. I am working with biological specimens doing standard thin section type stuff. are you doing some Material Sci application that demands more?
I will be interested in Alwyn (and any others) reply since I hope to buy one soon!
} . } } } There was a thread recently on scanners for TEM film. I have looked up } all the models mentioned, on the web and called agents for prices - and } produced a comparative table, given below. } } I do not guarantee that the figures are accurate but they are my best } interpretation of the data given. } } In the light of experience and Nestor's comments, I would suggest that } 2000 dpi is a minimum for TEM negatives. You may be able to get away } with less nine times out of ten, but there will be occasions when you } need more. } I would exclude the Minolta and all the Epsons from consideration } (despite the incredibly low prices of some of the Epsons) because of the } low pixel density. } } Among the rest the Nikon has the best pixel density and the best optical } density (another critical parameter for TEM negatives). The price is } very competitive too. The Nikon web site does not give a time for } scanning a negative. On the face of it the Nikon would be a best buy - } get a separate, inexpensive flatbed scanner for the other work. } } These comments are all my own opinions based on manufacturers' data. } Since we are considering purchase any comments to the contrary would be } most welcome. } } } } Code Maker Model Type } } } } A Agfa DuoScan T2500 Flatbed } -Transparency included } } B Epson 1640 several versions Flatbed } -Transparency option } 1680 several versions } } C 1600 several versions Flatbed } -Transparency included } } D Imacon Flextight Precision II Drum -for } film and large format } } E Minolta Dimage ScanMulti II Film } } F Nikon Super Coolscan 8000ED Film } } G Polaroid 45 Ultra Film } } H Umax Powerlook 3000 Flatbed } -Transparency included } } } } } } Code dpi OD Time Price } Opinion } at 6 x 9 cm } } } A 2500 x2500 3.4 3 min } $4,500 Fair } } B 1600 x 3200 3.6 } $300-$3000 Poor } } $800-$1400 Poor } } C 1600 x 3200 3.3 } $650-$1160 Not suitable } } D 2240 x2240** 3.9/4.1 N/A above } $10k Good: low pixel density } } E 1128 x 1128 3.6 } Not suitable } } F 4000 x 4000 4.2 N/A } $2,695 V. Good } } G 2500 x 2500 3.8 5 min } $7,495 Good but pricey } } H 3048 x 3048 3.6 3 min } $6,499 Good } } } -- } .......... } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University } 5 East Packer Avenue } Bethlehem } Pennsylvania 18015-3195 } Phone 610 758 4231 } Fax 610 758 4244 } jae5-at-lehigh.edu
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Noise is only part of the problem, or solution, when considering a quantitative approach to scanning in negative or positive images. Since noise in electronic detection systems is largely dependent on temperature, that must also be brought into consideration. But the characteristics of the detector can be even more important. An array type device, such as a CCD, can have characteristics that vary from pixel to pixel as sensitivity, dynamic range and noise susceptibility. Your desire for a Consumer's Report on scanners is probably most appropriate as these various effects are difficult, if not impossible, to measure in a lab. Even if possible, these measurements may not be extendable to similar machines that aren't individually tested. Best to try to find a consensus on visual traits.
On Tuesday, April 10, 2001 7:50 AM, Sinkler, Wharton [SMTP:WSinkler-at-uop.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Laurie and Gary, } } Would noise be a good criterion? Say, for a perfectly evenly darkened film } (if such a thing existed, or at least even on a scale { { collected pixel } size) - what is the value of the noise (standard deviation of pixel value) } as a function of film darkness (density)? } } This would presumably improve with the time of collection. Thus how "good" } your scanner is depends on how you run it or whether it lets you take a } slower scan or to average multiple scans. With the exception of drum } scanners these devices all use CCD arrays. So what is probably most of } interest is the signal to noise ratio as a function of illumination } intensity, with everything known about CCD's going into determining this. } The maximum density the scanner can handle is just the point at which the } noise swamps the signal. } } There must be some good literature out there on the sources of noise, } optimizing collection (scan) time etc. One article which might be a } starting point is: } } G. H. Campbell, W. E. King and D. Cohen "Analysis of Experimental Error in } High Resolution Electron Micrographs", Microscopy and Microanalysis vol. 3 } (1997) p. 451. } } This is not very detailed, and treats only the total random noise, i.e. } grouping noise arising in collecting the image with that arising from the } scanner. } } Now, finding a good "Consumer Report" test with hard numbers on commercial } models is likely to be a lot harder! } } Wharton } } } -----Original Message----- } } From: Gary Gaugler [SMTP:gary-at-gaugler.com] } } Sent: Monday, April 09, 2001 10:37 PM } } To: L. D. Marks } } Cc: MSA listserver } } Subject: Re: Scanners: quantitative accuracy } } } } -------------------------------------------------------------------- ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } How do you define "quantitative digitization?" i.e., what } } variables are you dealing with in this respect? What are } } the "absolute terms?" } } } } Anyone else have some ideas about this topic? } } } } gg } } } } } } At 10:10 AM 4/9/2001, you wrote: } } } } } I have been listening to the thread on scanners. Has anyone done } } } tests of how accurate they are in absolute terms for quantitative } } } digitization? } } } } } } ------------------------------------------------------- } } } Laurence Marks } } } Department of Materials Science and Engineering & } } } Center for Transportation Nanotechnology } } } Northwestern University } } } Tel: (847) 491-3996 Fax: (847) 491-7820 } } } mailto:ldm-at-risc4.numis.nwu.edu } } } http://www.numis.nwu.edu http://www.ctn.northwestern.edu } } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Question: My student accidently got immersion oil on the 40x Leica Plan Acromat. I took a chance and used a fine artist's brush and xylene to clean it recalling (I may be wrong) that lens adhesives are soluble in alcohols not xylene, toluene and similar. Well, the lens didn't fall out. Too bad because I need an excuse to upgrade! If (or when) this happens again, what would you recommend for cleaning?
Does anyone out there have any experience microtoming bone that has NOT been decalcified? Is it even possible? I'm hoping to be able to have an answer for the admin people--prior to simply trying it out and possibly damaging my diamond knife.
Don Moravits Senior Technician Southwest Research Institute 6220 Culebra Road San Antonio, Texas 78238
I am embedding feather barbs which are mostly spongey dry collagen with keratin and small air pockets.
I am attempting Jan Dycks method-
1. 0.25 M NaOH 30 mins 2. formic acid / absolute alcohol 2:3 2 hrs ( to fill air pockets with solution) 3. 15% epon / propylene oxide 3 days 4. standard graduated increase of epon / dehydrant
Any other suggestions? Possibly from other similar material such as plant material.
Thanks
Tim Quinn Kansas University Museum of Natural History Lawrence, KS 785-864-4556
I would love to take advantage of the Firewire option but my information is that the 8700 has a Dmax of 3.4 vs the 3.9 for the 1100. That is a significant difference. Do EM negatives of biological thin sections reach that? I think so. I do a lot of EM immunocytochemistry and have to look for gold (intensely black) against a very dark tissue component so I am hoping the higher Dmax improves my results. I frequently scan negatives on a Umax 1100 (Dmax 3.4??) and can't differentiate the gold from the background although by eye I can discriminate them when the negative is placed on a light box. Changing my exposure would give me an unuseable image for the rest of the tissue. Maybe this is an extreme case but I suspect that lots of "dark organelles" (e.g., lysosomes, nuclei) have fine structure that get lost in the scanning with a low Dmax scanner. Tom
} A colleague and I each recently bought Microtek scanners to scan TEM } negatives. } I have the Artixscan 1100 and he has the Model 8700 which has similar } characteristics (actually higher resolution -1200dpi), 3.9 dmax at } 42 bits color } (14 grayscale), and the glassless film carrier setup. The 8700 has USB and } Firewire interfaces and is cheaper ( {$1000), and the 1000 dpi Model 1100 has a } SCSI interface. You might want to check out the specs of the lower cost model } 8700 on the microtekusa website if your computer can handle USB or Firewire. } Both scanners have performed up to our expectations, which I would } characterize } as modest. Microtek does not supply a 3-1/4 x 4 " negative carrier } for standard } size TEM film but you can easily make a serviceable one from stiff paper or } light cardboard. } } How much scanner resolution should you buy? The answer depends on how you } intend to use it. Most applications do not require capturing the full } resolution of the negative. From a practical viewpoint, the scanner } resolution } just determines how many times you can magnify the negative image to } produce the } final print size. For example, to get a publication-size print at 300 dpi, an } image scanned at 1200 dpi scan could be zoomed 4X. A practical alternative to } spending more for higher scanning resolution is to take photos at higher } magnification. One exception is with lattice images from the TEM, which } (depending on the lattice fringe spacing on the negative) might require higher } scan resolutions to avoid getting a moire effect. (Of course, not everyone } agrees. My colleague prefers to always scan at the maximum resolution). } } What does a Dmax of 3.9 mean to you? To me it means a very dark } negative. D is } the log of the transmitted to incident intensity ratio. I wonder if } users ever } actually verify the manufacturer's specs with a calibrated density target. A } Dmax of 3.9 can be useful for scanning TEM diffraction patterns that } might have } high contrast, but TEM micrograph negatives of metals and ceramics generally } don't have that much contrast and biological thin section photos tend to have } rather weak contrast. If your negatives are simply dark, use shorter photo } exposure times. Scanning with maximum allowed grayscale resolutions (e.g., 14 } bits rather than 8) is highly recommended if you intend to enhance or adjust } images, but that's another story. } } } Larry Thomas } Pacific Northwest National Laboratory } MSIN P8-16 } P.O. Box 999 } Richland, WA 99352 } Phone: (509)372-0793 Fax: (509)376-6308 } Email: mailto:Larry.Thomas-at-pnl.gov } } } } ---------- } From: Tom Phillips } Sent: Tuesday, April 10, 2001 11:22 AM } To: Alwyn Eades } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: Scanners } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } I too am about to buy and I would make a couple of comments on your } evaluation. First, let me remind everyone that the Dynamic range is } a log scale so small numerical differences are significant. } } I also think the Nikon Coolscan 8000 looks great but it only takes a } 2.5 x 3.5 negative which is smaller than my JEOL and Hitachi EM } negative sizes (~ 3 1/2 by 4 1/2"). Have these EM manufacturers gone } to a smaller film size or is Nikon using a non-Japanese EM as their } standard? seems odd but I don't see how the Nikon would be very } useful. You say a {2000 line scanner would be useful 9 out of 10 } times but want the 2000+ lines for the occasional high res scan. I } would argue that the size of the negative was the more important } variable to be worried about. The Nikon couldn't handle 4x5 LM } negatives or transparencies from autoradiography of } Westerns/Northerns, etc. } } My leading candidate is the ArtixScan 1100 has a Dmax of 3.9 (about } $1600 with SCSI card). This was has a 1000 x 2000 dpi resolution. } more details at www.microtek.com. This is my leading candidate. It } was 4 negative carriers and I await word whether one could be } modified to carry a 3 1/2 by 4 1/2 negative. At worst, I will have } my scientific instrumentation shop guys fabricate a holder. It comes } with a glass 8 x 10 glass carrier for odd size negs but I want to } avoid Newton rings and want a glassless carrier. } } I would appreciate comments on the following argument (I think I have } this correctly figured out but am not sure since so many out there } seem to want to have a higher resolution scanner). I have a Fuji } Pictrography 3000 printer with a 400 dpi output that is as good as } any other widely available printer in the academic world. If you } figure the maximum published image size is about 8 inches, that would } mean the maximum image size be 3200 dpi wide. A 1000 dpi scan of my } negative would be 4500 x 3500 dpi. I could crop by about 28% or 10% } depending on the orientation of the negative and still be taking full } advantage of the printer resolution. In reality, most EM publication } prints are smaller than 8" wide so one could crop even more and still } not need more than 1000 dpi. A resolution } 1000 dpi would be } useful for subtle morphometric analysis but a 4000 dpi scan of a 3 x } 4 negative would be 192 MB. That is pretty big for doing morphometry } on! A 1000 dpi scan of a 3.5 x 4.5" negative would be about 16 MB } and that is much more manageable. Perhaps the difference is in the } type of EM we are doing. I am working with biological specimens } doing standard thin section type stuff. are you doing some Material } Sci application that demands more? } } } I will be interested in Alwyn (and any others) reply since I hope to } buy one soon! } } } } . } } } } } } There was a thread recently on scanners for TEM film. I } have looked up } } all the models mentioned, on the web and called agents for } prices - and } } produced a comparative table, given below. } } } } I do not guarantee that the figures are accurate but they are my best } } interpretation of the data given. } } } } In the light of experience and Nestor's comments, I would suggest that } } 2000 dpi is a minimum for TEM negatives. You may be able to get away } } with less nine times out of ten, but there will be occasions when you } } need more. } } I would exclude the Minolta and all the Epsons from consideration } } (despite the incredibly low prices of some of the Epsons) because of } the } } low pixel density. } } } } Among the rest the Nikon has the best pixel density and the best } optical } } density (another critical parameter for TEM negatives). The price is } } very competitive too. The Nikon web site does not give a time for } } scanning a negative. On the face of it the Nikon would be a best buy } - } } get a separate, inexpensive flatbed scanner for the other work. } } } } These comments are all my own opinions based on manufacturers' data. } } Since we are considering purchase any comments to the } contrary would be } } most welcome. } } } } } } } } Code Maker Model Type } } } } } } } } A Agfa DuoScan T2500 Flatbed } } -Transparency included } } } } B Epson 1640 several versions Flatbed } } -Transparency option } } 1680 several versions } } } } C 1600 several versions Flatbed } } -Transparency included } } } } D Imacon Flextight Precision II Drum -for } } film and large format } } } } E Minolta Dimage ScanMulti II Film } } } } F Nikon Super Coolscan 8000ED Film } } } } G Polaroid 45 Ultra Film } } } } H Umax Powerlook 3000 Flatbed } } -Transparency included } } } } } } } } } } } } Code dpi OD Time Price } } Opinion } } at 6 x 9 cm } } } } } } A 2500 x2500 3.4 3 min } } $4,500 Fair } } } } B 1600 x 3200 3.6 } } $300-$3000 Poor } } } } $800-$1400 Poor } } } } C 1600 x 3200 3.3 } } $650-$1160 Not suitable } } } } D 2240 x2240** 3.9/4.1 N/A above } } $10k Good: low pixel density } } } } E 1128 x 1128 3.6 } } Not suitable } } } } F 4000 x 4000 4.2 N/A } } $2,695 V. Good } } } } G 2500 x 2500 3.8 5 min } } $7,495 Good but pricey } } } } H 3048 x 3048 3.6 3 min } } $6,499 Good } } } } } } -- } } .......... } } Alwyn Eades } } Department of Materials Science and Engineering } } Lehigh University } } 5 East Packer Avenue } } Bethlehem } } Pennsylvania 18015-3195 } } Phone 610 758 4231 } } Fax 610 758 4244 } } jae5-at-lehigh.edu } } -- } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax)
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Does anyone know where I can find a multiple staining device for TEM grids using UA and Reynolds lead citrate that will result in CLEAN grids. Thank you Connie rosscac-at-okstate.edu
} Name: Kade } Organization: RMIT Melbourne Australia } Education: Graduate College } Location: Melbourne, Victoria, Australia } } Question: Hi, I have just purchased a microscope, a good biological } one. I need a microscope kit but nobody sells them around here. } Anyway already I have glass slides and covers. I have read some on } microscopy and I need an adhesive, resin I think its called to } prepare slides? is this true? } Also some ink to stain specimens. What are the names of all these } chemicals so I can buy them all seperatly since no one sells them all } together.
Kade -
You need a book on "microtechnique" plus a general idea of the types of specimens that you want to look at, BEFORE you start ordering things. Even tho you've "read some on microscopy " you could start with this book:
Nachtigall, W. 1995 Exploring With the Microscope 160 pp. hardbound. 6.5x9.5". $19.95. ISBN 0-8069-0866-1 Sterling Publishing Co., NY. Although this book is intended for adult amateur microscopists, it is well written and will provide teachers and classroom volunteers with much useful information on "serious" light microscopy. Almost half of the book is devoted to simple preparation methods for biological specimens and descriptions (with gool illustrations) of commonly encountered organisms. Adult. RECOMMENDED
You will find a lot of help on the amateur microscopy website http://www.microscopy-uk.org.uk
Both of these listings are from the Project MICRO bibliography (URL below).
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Your information is correct and mine is not. The Dmax of the 8700 is 3.4.
Larry
---------- From: Tom Phillips Sent: Wednesday, April 11, 2001 8:22 AM To: Microscopy-at-sparc5.microscopy.com Cc: jae5-at-lehigh.edu; Thomas, Larry (PNNL) Subject: RE: Scanners
I would love to take advantage of the Firewire option but my information is that the 8700 has a Dmax of 3.4 vs the 3.9 for the 1100. That is a significant difference. Do EM negatives of biological thin sections reach that? I think so. I do a lot of EM immunocytochemistry and have to look for gold (intensely black) against a very dark tissue component so I am hoping the higher Dmax improves my results. I frequently scan negatives on a Umax 1100 (Dmax 3.4??) and can't differentiate the gold from the background although by eye I can discriminate them when the negative is placed on a light box. Changing my exposure would give me an unuseable image for the rest of the tissue. Maybe this is an extreme case but I suspect that lots of "dark organelles" (e.g., lysosomes, nuclei) have fine structure that get lost in the scanning with a low Dmax scanner. Tom
} A colleague and I each recently bought Microtek scanners to scan TEM } negatives. } I have the Artixscan 1100 and he has the Model 8700 which has similar } characteristics (actually higher resolution -1200dpi), 3.9 dmax at } 42 bits color } (14 grayscale), and the glassless film carrier setup. The 8700 has USB and } Firewire interfaces and is cheaper ( {$1000), and the 1000 dpi Model 1100 has a } SCSI interface. You might want to check out the specs of the lower cost model } 8700 on the microtekusa website if your computer can handle USB or Firewire. } Both scanners have performed up to our expectations, which I would } characterize } as modest. Microtek does not supply a 3-1/4 x 4 " negative carrier } for standard } size TEM film but you can easily make a serviceable one from stiff paper or } light cardboard. } } How much scanner resolution should you buy? The answer depends on how you } intend to use it. Most applications do not require capturing the full } resolution of the negative. From a practical viewpoint, the scanner } resolution } just determines how many times you can magnify the negative image to } produce the } final print size. For example, to get a publication-size print at 300 dpi, an } image scanned at 1200 dpi scan could be zoomed 4X. A practical alternative to } spending more for higher scanning resolution is to take photos at higher } magnification. One exception is with lattice images from the TEM, which } (depending on the lattice fringe spacing on the negative) might require higher } scan resolutions to avoid getting a moire effect. (Of course, not everyone } agrees. My colleague prefers to always scan at the maximum resolution). } } What does a Dmax of 3.9 mean to you? To me it means a very dark } negative. D is } the log of the transmitted to incident intensity ratio. I wonder if } users ever } actually verify the manufacturer's specs with a calibrated density target. A } Dmax of 3.9 can be useful for scanning TEM diffraction patterns that } might have } high contrast, but TEM micrograph negatives of metals and ceramics generally } don't have that much contrast and biological thin section photos tend to have } rather weak contrast. If your negatives are simply dark, use shorter photo } exposure times. Scanning with maximum allowed grayscale resolutions (e.g., 14 } bits rather than 8) is highly recommended if you intend to enhance or adjust } images, but that's another story. } } } Larry Thomas } Pacific Northwest National Laboratory } MSIN P8-16 } P.O. Box 999 } Richland, WA 99352 } Phone: (509)372-0793 Fax: (509)376-6308 } Email: mailto:Larry.Thomas-at-pnl.gov } } } } ---------- } From: Tom Phillips } Sent: Tuesday, April 10, 2001 11:22 AM } To: Alwyn Eades } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: Scanners } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } I too am about to buy and I would make a couple of comments on your } evaluation. First, let me remind everyone that the Dynamic range is } a log scale so small numerical differences are significant. } } I also think the Nikon Coolscan 8000 looks great but it only takes a } 2.5 x 3.5 negative which is smaller than my JEOL and Hitachi EM } negative sizes (~ 3 1/2 by 4 1/2"). Have these EM manufacturers gone } to a smaller film size or is Nikon using a non-Japanese EM as their } standard? seems odd but I don't see how the Nikon would be very } useful. You say a {2000 line scanner would be useful 9 out of 10 } times but want the 2000+ lines for the occasional high res scan. I } would argue that the size of the negative was the more important } variable to be worried about. The Nikon couldn't handle 4x5 LM } negatives or transparencies from autoradiography of } Westerns/Northerns, etc. } } My leading candidate is the ArtixScan 1100 has a Dmax of 3.9 (about } $1600 with SCSI card). This was has a 1000 x 2000 dpi resolution. } more details at www.microtek.com. This is my leading candidate. It } was 4 negative carriers and I await word whether one could be } modified to carry a 3 1/2 by 4 1/2 negative. At worst, I will have } my scientific instrumentation shop guys fabricate a holder. It comes } with a glass 8 x 10 glass carrier for odd size negs but I want to } avoid Newton rings and want a glassless carrier. } } I would appreciate comments on the following argument (I think I have } this correctly figured out but am not sure since so many out there } seem to want to have a higher resolution scanner). I have a Fuji } Pictrography 3000 printer with a 400 dpi output that is as good as } any other widely available printer in the academic world. If you } figure the maximum published image size is about 8 inches, that would } mean the maximum image size be 3200 dpi wide. A 1000 dpi scan of my } negative would be 4500 x 3500 dpi. I could crop by about 28% or 10% } depending on the orientation of the negative and still be taking full } advantage of the printer resolution. In reality, most EM publication } prints are smaller than 8" wide so one could crop even more and still } not need more than 1000 dpi. A resolution } 1000 dpi would be } useful for subtle morphometric analysis but a 4000 dpi scan of a 3 x } 4 negative would be 192 MB. That is pretty big for doing morphometry } on! A 1000 dpi scan of a 3.5 x 4.5" negative would be about 16 MB } and that is much more manageable. Perhaps the difference is in the } type of EM we are doing. I am working with biological specimens } doing standard thin section type stuff. are you doing some Material } Sci application that demands more? } } } I will be interested in Alwyn (and any others) reply since I hope to } buy one soon! } } } } . } } } } } } There was a thread recently on scanners for TEM film. I } have looked up } } all the models mentioned, on the web and called agents for } prices - and } } produced a comparative table, given below. } } } } I do not guarantee that the figures are accurate but they are my best } } interpretation of the data given. } } } } In the light of experience and Nestor's comments, I would suggest that } } 2000 dpi is a minimum for TEM negatives. You may be able to get away } } with less nine times out of ten, but there will be occasions when you } } need more. } } I would exclude the Minolta and all the Epsons from consideration } } (despite the incredibly low prices of some of the Epsons) because of } the } } low pixel density. } } } } Among the rest the Nikon has the best pixel density and the best } optical } } density (another critical parameter for TEM negatives). The price is } } very competitive too. The Nikon web site does not give a time for } } scanning a negative. On the face of it the Nikon would be a best buy } - } } get a separate, inexpensive flatbed scanner for the other work. } } } } These comments are all my own opinions based on manufacturers' data. } } Since we are considering purchase any comments to the } contrary would be } } most welcome. } } } } } } } } Code Maker Model Type } } } } } } } } A Agfa DuoScan T2500 Flatbed } } -Transparency included } } } } B Epson 1640 several versions Flatbed } } -Transparency option } } 1680 several versions } } } } C 1600 several versions Flatbed } } -Transparency included } } } } D Imacon Flextight Precision II Drum -for } } film and large format } } } } E Minolta Dimage ScanMulti II Film } } } } F Nikon Super Coolscan 8000ED Film } } } } G Polaroid 45 Ultra Film } } } } H Umax Powerlook 3000 Flatbed } } -Transparency included } } } } } } } } } } } } Code dpi OD Time Price } } Opinion } } at 6 x 9 cm } } } } } } A 2500 x2500 3.4 3 min } } $4,500 Fair } } } } B 1600 x 3200 3.6 } } $300-$3000 Poor } } } } $800-$1400 Poor } } } } C 1600 x 3200 3.3 } } $650-$1160 Not suitable } } } } D 2240 x2240** 3.9/4.1 N/A above } } $10k Good: low pixel density } } } } E 1128 x 1128 3.6 } } Not suitable } } } } F 4000 x 4000 4.2 N/A } } $2,695 V. Good } } } } G 2500 x 2500 3.8 5 min } } $7,495 Good but pricey } } } } H 3048 x 3048 3.6 3 min } } $6,499 Good } } } } } } -- } } .......... } } Alwyn Eades } } Department of Materials Science and Engineering } } Lehigh University } } 5 East Packer Avenue } } Bethlehem } } Pennsylvania 18015-3195 } } Phone 610 758 4231 } } Fax 610 758 4244 } } jae5-at-lehigh.edu } } -- } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax)
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Dear all, We are having a great deal of difficulty getting hold of our normal film "Tritiated Hyper film" for use with tritiated ligands. Does any one out there know of a local supplier (U.K.)
We use 70 % isopropanol to clean emersion oil off of our lenses.
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan (734)763-1170 FAX (734)763-1166 dsoren-at-umich.edu
On Wed, 11 Apr 2001 mckaylodge-at-aol.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Email: mckaylodge-at-aol.com } Name: Robert Lodge } } Organization: McKay Lodge Home School } } Education: 9-12th Grade High School } } Location: Oberlin, OH 44074 } } Question: My student accidently got immersion oil on the 40x Leica } Plan Acromat. I took a chance and used a fine artist's brush and } xylene to clean it recalling (I may be wrong) that lens adhesives are } soluble in alcohols not xylene, toluene and similar. Well, the lens } didn't fall out. Too bad because I need an excuse to upgrade! If (or } when) this happens again, what would you recommend for cleaning? } } Bob Lodge } } --------------------------------------------------------------------------- }
Has anyone had experience in determining the concentration of particles in a solution by counting on EM grids in a similar way to using a haemocytometer? Is it possible to count the particles in a defined number of grid squares then calculate back to the area of the grid and the amount of solution which was applied and allowed to dry down?
Richard Gardiner Department of Plant Sciences University of Western Ontario
Just tried to order some Kodabrome II RC paper, grade F5. Vendor says it is discontinued. Any suggestions for an alternative? We have been using Kodabrome RC II for a long time with a Mohr processor. How about Polycontrast RC? Or is this the beginning of the end for old fashioned darkrooms?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
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One of the big advantages of computer printing of TEM images is the ability to adjust the contrast of the final image. However, to prevent generating a negative that has completely overexposed areas which are difficult to salvage by any means-such as one may get with metal or ceramic specimens-an alternative film developer may help. There are two bath "split" developers which lower contrast to manageable levels, in effect chemically "dodging" a developing negative. Fixing is carried out normally. I'm told biological specimens don't usually need such treatment. I have no financial interest in the following company, but I am pleased with the results obtained with their developer called Diafine. It is made by Acufine Inc., 5441 North Kedzie Ave. Chicago, Il., 60625.
Bernard Kestel E-mail: {kestel-at-anl.gov} Materials Science Division Argonne National Laboratory Argonne, Il., 60439
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id TAA26206 for dist-Microscopy; Wed, 11 Apr 2001 19:12:49 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id TAA26197 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 11 Apr 2001 19:12:19 -0500 (CDT) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id TAA26190 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 11 Apr 2001 19:12:08 -0500 (CDT) Mime-Version: 1.0 X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {p05001900b6faa35d2003-at-[206.69.208.21]}
Below is the result of your feedback form. It was submitted by (jgh7f0-at-mizzou.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, April 11, 2001 at 14:32:21 ---------------------------------------------------------------------------
Email: jgh7f0-at-mizzou.edu Name: john harris
Organization: university of missouri-columbia
Education: Undergraduate College
Location: Columbia, Missouri
Question: Dear Sir or Madam; i am looking for an image of S. Agalactiae attached to an epithelial cell or some other gram + cocci.
Kodabrome is for sure an old product. But a good one. I quit using it long ago in favor of Ilford paper. Ilford makes RC paper--but my work has only been output to archival fiber paper. But I suspect that the RC quality is still up to par.
I use plain matte for normal prints--and archival matte fiber for fine art prints (nudes, etc.). I use a Kreonite processor for the print papers. All b/w neg media is hand processed in separate roll holders (Nikor). My normal format is 6x4.5cm and 6x7cm. The same recipes would apply to other formats.
I did some 4x5" work in prior years and do the same mechanics today.
Try some Ilford paper.
gary g.
http://photoweb.net
At 03:25 PM 4/11/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Wow...this is really deja vu. I used Diafine and Acufine way many years ago. At that time, the issue was push processing. Today, the zone system prevails. Using the zone system does not require special chemicals. It is a matter of how the neg is rated and how it is processed.
Look up some of Ansel Adams' works (The Negative). The main idea is to adjust your neg's EV for total tonal range such that it can be printed with less than bone crushing effort.
All of my fine art prints are shot and printed using this process.
gary g.
http://photoweb.net
At 04:41 PM 4/11/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I used Illiford RC paper for a long time and I liked it better than anything else I have ever used. It may be just be personal preference but it seemed to have brighter higlights and blacker blacks than I could get with other RC papers. Just be sure and get Illford filters to go with it. One other nice thing is it has a matt finsh as well as glossy if you don't want glossy prints. The matt will scan great. It doesn't have texture problems like most pearl or simi-glossy papers have. The matt finish displays a lot better than glossy finsh IMHO.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
} From: "Gary Gaugler" {gary-at-gaugler.com}
} Kodabrome is for sure an old product. But a good one. } I quit using it long ago in favor of Ilford paper. Ilford } makes RC paper--but my work has only been output to archival } fiber paper. But I suspect that the RC quality is still up to par. } } I use plain matte for normal prints--and archival matte fiber } for fine art prints (nudes, etc.). I use a Kreonite processor } for the print papers. All b/w neg media is hand processed } in separate roll holders (Nikor). My normal format is 6x4.5cm } and 6x7cm. The same recipes would apply to other formats. } } I did some 4x5" work in prior years and do the same } mechanics today. } } Try some Ilford paper. } } gary g. } } http://photoweb.net } } } } } Just tried to order some Kodabrome II RC paper, grade F5. Vendor says it is } } discontinued. Any suggestions for an alternative? We have been using } } Kodabrome RC II for a long time with a Mohr processor. How about } } Polycontrast RC? Or is this the beginning of the end for old fashioned } } darkrooms? } } } } Jonathan Krupp
} I am using Polymax II RC. For draft pictures it's good enough in my point } of view. In compare with Ilford Multigrade III RC, Polymax gives better } contrast (filter #5 on Ilford is equal to #3 on Polymax in my hands).
Sergey
} At 03:25 PM 4/11/2001, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Usualy to reduce contrast you under expose the film. Then develop it to the disired density using a developer that generates low contrast. One way to reduce contrast is to dilute you developer by a factor of 2, 4 or more with water. It extends the developing time a good deal but it reduces the contrast. You might also look at low contrast developers that work with the film you are using.
A few question on rec.photo.darkroom will get you more information than you can handle and some of it will actually work.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
} } } } } } One of the big advantages of computer printing of TEM images is the } } ability to adjust the contrast of the final image. However, to prevent } } generating a negative that has completely overexposed areas which are } } difficult to salvage by any means-such as one may get with metal or ceramic } } specimens-an alternative film developer may help. } } There are two bath "split" developers which lower contrast to } } manageable levels, in effect chemically "dodging" a developing negative. } } Fixing is } } carried out normally. I'm told biological specimens don't usually need } } such treatment. } } I have no financial interest in the following company, but I am } } pleased with the results obtained with their developer called Diafine. It is } } made by Acufine Inc., 5441 North Kedzie Ave. Chicago, Il., 60625. } } } } Bernard Kestel E-mail: {kestel-at-anl.gov} } } Materials Science Division } } Argonne National Laboratory } } Argonne, Il., 60439 } }
-----Original Message----- } From: christine [mailto:ac.richardson2-at-btinternet.com] Sent: Wednesday, April 11, 2001 1:59 PM To: microscopy-at-sparc5.microscopy.com
Dear all, We are having a great deal of difficulty getting hold of our normal film "Tritiated Hyper film" for use with tritiated ligands. Does any one out there know of a local supplier (U.K.)
Back in the pre PC (chemical) days, toluene was a recommended solvent for cleaning lenses of immersion oil. Now, we at National Steel, wipe off excess oil with lens tissue and then clean the lens with Kodak lens cleaner solution.
Best regards,
Sam Purdy National Steel Tech Center Trenton MI
} ---------- } From: mckaylodge-at-aol.com } Sent: Wednesday, April 11, 2001 9:59 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ask-A-Microscopist:Help Cleaning Lenses } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Email: mckaylodge-at-aol.com } Name: Robert Lodge } } Organization: McKay Lodge Home School } } Education: 9-12th Grade High School } } Location: Oberlin, OH 44074 } } Question: My student accidently got immersion oil on the 40x Leica } Plan Acromat. I took a chance and used a fine artist's brush and } xylene to clean it recalling (I may be wrong) that lens adhesives are } soluble in alcohols not xylene, toluene and similar. Well, the lens } didn't fall out. Too bad because I need an excuse to upgrade! If (or } when) this happens again, what would you recommend for cleaning? } } Bob Lodge } } -------------------------------------------------------------------------- } - }
Kodabrome has not been discontinued across the board. However, certain size/surface/grade combinations have been. Kodabrome II RC F5 is available in 250sheet 8x10 packages only. We stock them but a dealer should be able to order them. As far as alternatives.... Agfa offers Brovira Speed RC, a developer incorporated paper in grades 2-5 and available in 100 sheet 8x10 packs. Brovira Speed is a cold tone paper. Agfa also offers a Multicontrast RC product. From Kodak, Polycontrast or Polymax are variable contrast papers. Polycontrast is developer incorporated, as Kodabrome is, so it will process in developers as well as many activators. Polymax requires the use of a developer and will not process in activators. Ilford offers Multigrade IV Deluxe which is not developer incorporated. All of these papers use filters to control contrast, although a #5 filter with any of them is not the same as a grade 5 Kodabrome. Both Polymax and Multigrade IV have a slightly wider contrast range than Polycontrast. For those who prefer a cooler tone print, Ilford also has a new paper, Multigrade Cooltone, a non developer incorporated paper with a cooler image tone and a cool white base tint.
George
George Laing National Graphic Supply v:(800) 223-7130 X3109 f:(800) 832-2205 email: scisales-at-ngscorp.com
} Just tried to order some Kodabrome II RC paper, grade F5. Vendor } says it is discontinued. Any suggestions for an alternative? We have } been using Kodabrome RC II for a long time with a Mohr processor. How } about Polycontrast RC? Or is this the beginning of the end for old } fashioned darkrooms? } } Jonathan Krupp
The paper is still listed on the Kodak Web sight, http://www.kodak.com/cluster/global/en/professional/support/techPubs/f33/old/f33.shtml . The web sight does list Kodabromide paper as discontinued but lists Kodabrome II RC as its replacement and does not list the latter as discontinued. I was curious, so, I called the 800 technical hotline (800) 242-2424 (option 02 gives you an operator) and asked. They said that it is still available but not in as many sizes as it use to be.
I've had venders tell me a product is discontinued when it is only discontinued from their stocking process, not by the manufacture. In this case it may be that only the size you wanted was discontinued. You may want to call the vendor back or check with another vender.
I have no connection with Kodak but wanted to see if the predictions I'd heard about, as you put it, " the beginning of the end for old fashioned darkrooms," was starting. It would seem not quite yet in this case. Though I'm hearing that in 2 or 3 years it will, somewhat sad I think.
Jim Roberts
James L. Roberts Firearm and Toolmark Examiner Ventura Co. Sheriff's Lab (805) 654-2308 James.Roberts-at-mail.co.ventura.ca.us
} } } {jmkrupp-at-cats.ucsc.edu} 04/11/01 03:25PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi:
Just tried to order some Kodabrome II RC paper, grade F5. Vendor says it is discontinued. Any suggestions for an alternative? We have been using Kodabrome RC II for a long time with a Mohr processor. How about Polycontrast RC? Or is this the beginning of the end for old fashioned darkrooms?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Yes, Thermanox cover slips survive treatment with HMDS. We occationally process cell monlayers grown on them for SEM.
Regards,
Dotty
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan (734)763-1170 FAX (734)763-1166 dsoren-at-umich.edu
On Thu, 12 Apr 2001, Randall, Kevin J wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I need to process some thermanox coverslips for SEM. Does anyone know } whether thermanox survives HMDS? } } Cheers } } Kevin }
Hi All: Thank you all for sharing your experiences about working with SiC. Your suggestions have helped me come up with a plan of attack. Regards, Mike Coviello UT Arlington
We are a Kodak distributor and we checked with Kodak this morning and Kodabrome II RC paper, grade F5 is still available. If you still interested you may contact me off-line.
John Arnott
Jon Krupp wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Hi: } } Just tried to order some Kodabrome II RC paper, grade F5. Vendor says it is } discontinued. Any suggestions for an alternative? We have been using } Kodabrome RC II for a long time with a Mohr processor. How about } Polycontrast RC? Or is this the beginning of the end for old fashioned } darkrooms? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
We are about to install a Kimball ES-423E (extended life) LaB6 cathode in a Hitachi H7100 TEM and were wondering if anyone had some starting points for the voltage settings for filament heating. This would save us a lot of time, if so.
Thank you.
John B.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 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 ##############################################################
Here is an odd way that I thought up to clean oil from an objective without using solvents that may attack lens cement.
Remove the lens from the scope. I usually like to view the oil contamination using a stereoscopic microscope. Wipe excess oil away - I like using Ross Optical Paper. Using a cotton applicator wrapped in Ross optical paper, apply a small amount of Dawn dishwashing detergent. Gently work the surface to emulsify the oil into the detergent using the optical papered applicator. You may have to add a small amount of water. Do not allow fluids to go much beyond the lens area!
At this point, hold the lens vertical with the back focal plane pointing up. Apply a small amount of deionized water on the final lens element from the side of the opjective to create a hanging drop. I usually use a wash bottle or 10cc syringe. The surface tension of water will create a small drop around the optical surface. Start a stream of DI water flowing through the small drop to wash away the oil/water suspension. After about a minute, stop the water flow while allowing a small drop of water to stay on lens. At this time, blow the water off using C02 or some other clean compressed gas. This will eliminate streaks. Upon inspection, if you see contamination, repeat the process and your problem should be solved.
It's weird but it works.
Robert
Robert Fitton Teaching Associate/Director of Laboratories Luther College Department of Biology 700 College Drive Decorah, IA 52101
Voice 563-387-1559 FAX 563-387-1080
Enjoy a visit to our website: http://www.luther.edu/~biodept/
Dear Richard, The most common use of this is for asbestos fibre load determination. I did it once. You count the number of fibres in a specified number of grids openings, then calculate back to the original collected volume or vacuumed area. It should work similarly for any recognizable particle. I could look up the method, if you like. At 06:08 PM 4/11/01 -0400, you wrote: } } Has anyone had experience in determining the concentration of particles } in a solution by counting on EM grids in a similar way to using a } haemocytometer? Is it possible to count the particles in a defined } number of grid squares then calculate back to the area of the grid and } the amount of solution which was applied and allowed to dry down? } } Richard Gardiner
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Below is the result of your feedback form. It was submitted by (dngeorge-at-wam.umd.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, April 12, 2001 at 16:04:35 ---------------------------------------------------------------------------
Email: dngeorge-at-wam.umd.edu Name: Damali Martin
Organization: University of Maryland
Education: Graduate College
Location: College Park, MD
Question: I have embedded and sectioned some salivary glands in epon and have placed the sections on glass slides. However, when I counterstain, a high percentage of sections float off and are lost. How can this problem be prevented?
Also, several of my sections have wrinkles in them. Is there any trick to getting rid of them?
Anyone got any suggestions as to the cause of (and remedy for) the pronounced image shift that occurs when I turn the Y stigmator control on my JEOL 840A?
It also has the usual stigmatic effect.
The X control shifts the image only very little.
The coils seem to check out OK.
There's something quite poetic about working on this particular part of the instrument today of all days.
thanks
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Stigmators operate as two opposing coils. It may be that one of the coils for the Y stigmator is not active while the opposing Y coil is. Perhaps one of the coils has shorted? The resistance of these coils is very low, making a simple check rather difficult, not to mention that they are often series connected within the column making physical access difficult. But if you can measure the current drawn by each, you may find a significant difference if one is shorted.
If so, the only remedy is replacement of the coils.
On Friday, April 13, 2001 1:12 PM, Ritchie Sims [SMTP:r.sims-at-auckland.ac.nz] wrote: } } Hi } } Anyone got any suggestions as to the cause of (and remedy for) the } pronounced image shift that occurs when I turn the Y stigmator } control on my JEOL 840A? } } It also has the usual stigmatic effect. } } The X control shifts the image only very little. } } The coils seem to check out OK. } } There's something quite poetic about working on this particular part } of the instrument today of all days. } } thanks } } rtch } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
About 25 years ago, when I was taking the embryology course at the Marine Biology Laboratory at Woods Hole, we were taught the following methods for cleaning oil from lenses by Robert Allen, a well known microscopist:
Remove the lens from the microscope and invert it. Remove most of the excess oil from the objective with lens paper without touching the objective surface itself. Gently lay a strip of lens paper over the objective. Since the lens is recessed the lens paper does not touch the glass directly. Then place a drop or two of ether on the lens paper just next to where it contacts the lens, and draw the lens paper over the objective surface. The ether vapors swirl around under the lens paper and dissolve the oil while the lens paper absorbs the mixture and carries it away, without actually touching the glass surface. You might need to repeat a few times to remove the oil.
It works with all but the most stubborn deposits. The upside is that it completely avoids touching the glass surface, and the ether fumes are in contact with the element such a short time that it miakes it less likely you will dissolve the glues that hold the lens elements together. The downside is that it requires ether. I haven't tried it with other solvents. -- Gary P. Radice gradice-at-richmond.edu Associate Professor of Biology 804 289 8107 (voice) University of Richmond 804 289 8233 (FAX) Richmond VA 23173 http://www.science.richmond.edu/~radice
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Accountabilities % of time
70 Collect data from production activities to track and improve quality. Develop collection systems to optimize this process. We have a new Hitachi S3000N with an Oxford INCA EDS system and an Oxford/Gatan CL system with cryostage. Routine analysis will include EBIC, EDS, and CL.
20 Interface with Manufacturing Engineers, Application engineers, Detail Engineers, Product Managers, Technicians, Maintenance, Tool Room, Field Service, and other stakeholders to achieve assigned project goals. Recommend design changes to improve manufacturability and quality.
10 Coordinate project work activity, including testing, machining, and purchasing, as needed to ensure meeting project schedules. Design and build fixtures, prototypes, and samples. Provide training on equipment and techniques to assemblers.
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Learn about ADC - The Broadband Company at www.adc.com
I usually see image shift as a result of stigmation in both of our scopes (JEOL 840A and Hitachi 2460N). I rather accepted that it just went with the territory. If it is a sign of a problem with the scope or its alignment, I too would be interested in hearing how to eliminate it.
Warren
At 06:12 PM 4/13/2001 +0000, you wrote:
} Hi } } Anyone got any suggestions as to the cause of (and remedy for) the } pronounced image shift that occurs when I turn the Y stigmator } control on my JEOL 840A? } } It also has the usual stigmatic effect. } } The X control shifts the image only very little. } } The coils seem to check out OK. } } There's something quite poetic about working on this particular part } of the instrument today of all days. } } thanks } } rtch } } Ritchie Sims Phone : 64 9 3737599 ext 7713
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
The 840 has some pots (potentiometers) on one of the circuit boards that adjust out the image shift while adjusting the stigmators. They probably balance a bias, or an offset, in the circuit for the stigmation coils. If you have the manual with the schematics, they may be identified. I may be able to find it in ours, if you don't have it.
Normally, X or Y stig adjustment will cause image shifting. More shifting at higher mag. Since stigmation is done with coils by the scan coils, changing current in the stig coils appears to the beam as a change in scan coil current. Hence, the image shifts. Later day systems account for this by feeding some of the stig signal to the scan coils as feedback. It will also accept additional feedback from magnification. The operation is to adjust scan coil current and condenser current as stig is adjusted so that the net result is minimal image shift with varying stig input.
Since one stig channel out of two is not working right, there are likely one of two or three things which could go wrong. First, and worst, the stig coil is open or shorted. Somewhat unlikely I think. Second, one part of the stig coil drive circuit has failed. Third, some part of the feedback system has failed. Since you say that it does stig, but causes image shift, then the stig circuit itself and the coil is OK. You can verify this.
Since the stig coil drive systems are identical (ususally), you should be able to trace readings from the good channel and compare them to the bad channel. This should show right away where the problem is. The stig, beam alignment and image shift circuits are usually all the same. If so in your system, they will give plenty of data points for checking. The coils (one for X, one for Y) are typically driven by push-pull power transistors which are high current buffers on the output of a small op amp. The coils are in the negative feedback loop of the op amp. One lead of a coil would connect to the output of the buffer transistors (large ones) while the other end connects to the inverting input of the op amp and is returned to ground through a low value resistor. Measuring the voltage across this resistor will tell how much current is flowing through the coil (I=E/R).
Stig effect is lower at lower mags. So there would be less automatic compensation for stig vs. mag. Try a low mag setting and measure the voltage on each side of the stig coils (two leads each) and the voltage on the low value resistors. Have the stig controls at 12 O'clock each. If these readings match, odds are that the coils are for sure OK. Then the problem ought to be narrowed to the stig balance circuit. Look for this circuit and compare the two stig signal feedback loops for differences. It could be something as simple as a bad op amp in the path from the stig circuit to the beam alignment coil drivers. Since stig has little effect at low mag, but huge effect at high mag, the usual control path for stig compensaton versus magnification is to electronically change the beam position by sending a stig sourced voltage to the scan coil driver circuit.
If you don't have schematics for the system, that is of course a major problem. In this case, perhaps someone who has your same model has encountered this problem before and knows of the failure mechanism and cause.
gary g.
At 11:12 AM 4/13/2001, you wrote:
} Hi } } Anyone got any suggestions as to the cause of (and remedy for) the } pronounced image shift that occurs when I turn the Y stigmator } control on my JEOL 840A? } } It also has the usual stigmatic effect. } } The X control shifts the image only very little. } } The coils seem to check out OK. } } There's something quite poetic about working on this particular part } of the instrument today of all days. } } thanks } } rtch } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
Stigmator shift is almost always down to the balance of the stigmator coils. Stigmators have 8 coils in four sets of two. If an opposing pair are not correctly balanced the stronger coil will cause the image to move away from that coil.
Some instruments have the balancing potentiometers easily accessible others hide them away in the electronics.
The pots will be called Xx, Xy, and Yx, Yy I do not know where they are on a JEOL 840 but if you look inside you may be lucky?
To adjust - 1. Place an easily recognizable feature in the center of the screen 2. Turn the X stigmator fully in one direction and re center with the Xx or Xy control whichever fits. 3. Turn in the opposite direction and use the other compensation control to center 4. Repeat with the Y stigmator and Yx and Yy controls.
I have just completed the promised "Monitoring & Maintaining Electron Microscope Performance" interactive CD, this area is covered in the instrument tuning section so its pretty fresh in my mind. Need to see even more for yourself then we have the course of the same name running in Sydney early October this year?
Kindest regards and good luck
Steve Chapman Senior Consultant, Protrain For professional training in SEM, TEM and EDX world wide www.emcourses.com
I have a Hitachi SEM model S-570 (1985). I am interested of getting a X-ray analyzer for this microscope. My preference would be to buy a used analyzer with an ultrathin window for light elements analysis.
If you know someone who is interested in selling it's analyzer, please let me know...
Thank you
Christian Normand Tekna Systems Plasma (819) 820-2204
I hesitated to make any response regarding this subject. But--here goes anyway. Someone may find it useful.
I think that the issue is not reduction of contrast but rather extension of tonal range of the negative. That is what the zone system is all about. I think that it is a fact that if the silver is not exposed, there is not going to be any information gleaned from that unexposed or too underexposed area. The approach I use for fine art images is to overexpose and under-develop. It takes a LOT of work to develop a complete system of ISO rating, development and printing to achieve marketable prints. But it certainly can be done. The goal is to minimize the time spent in the darkroom doing printing. Ideally, the neg should print without any effort on grade 3 paper. I only use Ilford archival matte fiber paper, so extensions to RC papers may or may not directly apply. I also do not use variable contrast paper. But the principles are extensible I would think.
Being all-digital with my SEM, I use realtime histogram feedback to adjust gain and dynamic range. For TEM, I have no direct experience. But I might suggest that the same photo techniques for fine art neg work may apply to TEM negs. If you think that this might be true, read on. Otherwise, nevermind.
A neg with bad dynamic range (huge extremes of EV or low contrast) is not going to produce a very good scan. This is the realm of drum scanners with 4.0-6.0D. Pixel resolution is subordinate to D range in this case. If one is going to print a neg on paper, that is one aspect of the problem. If the neg is to be output to a magazine or printed page, that is another aspect. So the crux of the matter is what the actual intended end use of the neg is to be? If it is a nice print, OK. If it is a magazine or litho output, these are 133lpi. Doing scans at 4000dpi only to print them in a magazine at 133lpi is rather silly. One would probably be better off just printing the neg and scanning at 300dpi.
But the 300 dpi is the effective dpi for the size of the final image....not the original neg. So, if the neg is say 3" x 4" and is to be printed at that same size, a 300 dpi scanner should do the job (ignoring tonal range for the time being). If the neg is to be printed at twice its physical size, then the scanner has to have twice the resolution (600 dpi). And it goes on from there. Thus, for most image output methods on a printed page, 600-1200 optical dpi should be plenty of resolution. If you agree with this discussion at this point, then the resolution factor should no longer be an issue. What remains is D range.
(Note that commercial images are created at high resolution to accommodate manipulation and merging with other images. This way, image quality can gradually be reduced without affecting the final result.)
But the negative is the key item in all of this discussion. A bad neg is not likely to do anyone much good. So the challenge is to get a good neg at the get go. Here we go, back to the zone system. The idea is to use the neg as a non-linear image capture media for data which may span a wide dynamic range. And in so doing, be able to output (print or scan) this information with fidelity and minimal effort. This drives D value.
An unexposed, developed neg will have a transmission of 100%. It is actually slightly lower due to absorption by the medium, emulsion and residual anti-dispersion coating. But at 100% transmission, the neg would have an opacity of 100%input/100%output or 1.0. Since D=log1/T, the D value for the unexposed neg is log 1/1=0.0. This then is Dmin. The area with the most exposure (darkest region) will pass the least amount of transmitted light. If, for example, this region passes 1% of the transmitted light, this is a Tval=100%/1%=100. And D=log (1/100)=2.0. This then is Dmax. The D range of the negative is Dmax-Dmin=2.0-0.0=2.0. So a scanner with a D value of } 2.0 would capture the tonal range of this neg. The tonal range of this neg is 100 tonal variations. Prints generally have D values between 1.7 and 2.0. So the same scanner would handle the neg and a typical print.
Let's say that the darkest area of the neg transmits 0.1% of the transmitted source light. This then yields Tval=100/0.1 =1000. And D=log 1000 = 3.0. Thus, this neg has 1,000 tonal variations in it from clear to black. If the neg has double the number of tonal variations (2,000) that would mean that the opacity is 2,000 and D=log 2000 = 3.3. At 4000 tonal range, D=3.6. Thus, each 0.3D equates to doubling the opacity range or halving or doubling the transmission value. Thus, if a scanner has a D rating of 3.0 (1000 tonal ranges) and the neg passes a corresponding 0.1% of the transmitted light, and another scanner has a D rating of 3.3 (2000 tonal ranges) and the neg passes 0.05% of the transmitted light--can you really tell one from the other?
The eye is more responsive to subtle changes in tonal variations in the white region of an image than in the dark ones. Thus, it is important to retain as much variability and rendition in the darkest areas of the negs (the highlights) but to do so without sacrificing detail in the shadows (clearer areas of the neg). This is done using the zone system and expansion and contraction of tonal range.
The response of the neg emulsion's exposure to light is not linear over the range of clear to full black. Its response curve is somewhat like a flattened S. The low exposure region is called the toe, which consists of the film base + fog density (Dmin). Moving up the curve is the straight line section (not exactly a straight line), and then to the shoulder. At this point, increasing amounts of light do not have corresponding quantitative amounts of change in density. Further exposure reaches saturation, or Dmax. At this point, no increase in exposure will change the density of the neg.
The ultimate goal is to maximize the straight line section and move Dmax as high as possible, without detrimentally affecting Dmin. I use contraction to accomplish this. The approach is to overexpose and underdevelop. With a neg, the rule of thumb is to expose for the shadows. If there is insufficient exposure in the shadow areas, there will not be any detail rendered.
Using Ilford FP4+ film, I rate it at ISO 80 (mfg=125). Development is done using stock developer diluted 1:1. It is used one time and discarded. Never replenish or try to refresh the developer. Development time will vary, depending on the tonal range of the scene. Here are some extreme examples of how this system works: (if you are offended by nude images, use the still life links. These following links are to fine art nudes and still life which were done using the zone system previously described)
[nudes] A. This shot illustrates a typical impossible shot. Inside the room, the exposure was about EV 3, the outside was dense fog with a starch white picket fence at EV 11. A scene like this with eight stops of variation would typically be shot to either render detail in the highlights (fog and fence) or in the inside room's details (shadows). To render shadow detail and retain highlight detail, the zone system was used as described. The image via this link is as-scanned on a UMAX Powerlook III of a 6x6cm negative using the transparency adapter. You can see the detail in the paint on the wall and can see the fence in the fog. http://www.photoweb.net/pw_gal_nude/pw_gal_nude_1/g_2.html
B. This shot shows great shadow detail despite the outside light creating a hot spot on the model's head. And the window on the left was rendered, despite the high level of light. http://www.photoweb.net/pw_gal_nude/pw_gal_nude_1/g_6.html
C. This shot shows a rather low contrast scene. There are no remarkable highlights. There is much material in shadow. Overexposure and ensuring at least two zones of exposure for the shadows and then overdeveloping N+1 achieved a perfectly printable neg. Again, this neg is shown as-scanned. http://www.photoweb.net/pw_gal_nude/pw_gal_nude_6/g_3.html
[Still life] A. high side lighting. http://www.photoweb.net/pw_gal_still/pw_gal_still_4/g_5.html
B. Low shadow lighting, highlights present. http://www.photoweb.net/pw_gal_still/pw_gal_still_4/g_6.html
C. Even lighting. There is actually more detail revealed than is showed in this web pix. http://www.photoweb.net/pw_gal_still/pw_gal_still_3/g_6.html
Non-web images of these negs are of course much better than those on-line. But I hope these illustrate the points of the zone system. I do think it can apply to TEM negs. The other point is to keep in mind the relationship of D values of scanners to what you are actually going to be scanning. If a scanner has sufficient resolution, and say a D rating of 3.2. Are you really going to be able to tell any difference using a scanner with a 3.4 or 3.5D at much higher cost? If you have a densitometer, check the D range of some of your negs. They are probably all less than 3.0. Maybe I'm wrong in this respect since I have little experience with TEM media. But the idea is go get the equipment you need for the job you need (the output destination and the use of the image) based on the actual media being scanned. Otherwise, there is a great opportunity to buy capability which will never be utilized.
Despite all the discussion of processing negs, as more digital capture and image processing products come out, things will change. There are rather simple ways to expand the contrast of an otherwise low contrast, poor neg. Likewise, there are ways to extract subtle detail from negs which have blown out highlights. More about this later.
gary g.
Reference: Adams, A. (1981). The negative. Boston: Little, Brown and Company. ISBN 0-8212-1131-5 (twelfth printing, 1992).
At 01:07 AM 4/12/2001, you wrote:
} Usualy to reduce contrast you under expose the film. Then develop it to } the disired density using a developer that generates low contrast. One way } to reduce contrast is to dilute you developer by a factor of 2, 4 or more } with water. It extends the developing time a good deal but it reduces the } contrast. You might also look at low contrast developers that work with } the film you are using. } } A few question on rec.photo.darkroom will get you more information than } you can handle and some of it will actually work. } } Gordon } Gordon Couger gcouger-at-couger.com } Stillwater, OK www.couger.com/gcouger
Good job, Gary! Gary's description of dynamic range is the clearest and most easily understood I have seen (and I have been looking for a good one!). I think I already understood most of what he said before he said it but I was having a devil of a time trying to articulate the concept to one of my staff. My only followup question is in regards to his statement near the end where he suggests some test the density of a TEM negative with a densitometer to see if they really go much above 3.0. I would like to encourage any one who has or will be measuring this for a typical and even finicky biological thin section TEM image to please post the info on the Microscopy listserver. Thanks.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I hesitated to make any response regarding this subject. } But--here goes anyway. Someone may find it useful. } } I think that the issue is not reduction of contrast but rather extension } of tonal range of the negative. That is what the zone system is } all about. I think that it is a fact that if the silver is not exposed, } there is not going to be any information gleaned from that } unexposed or too underexposed area. The approach I use for fine } art images is to overexpose and under-develop. It takes a LOT } of work to develop a complete system of ISO rating, development } and printing to achieve marketable prints. But it certainly can } be done. The goal is to minimize the time spent in the darkroom } doing printing. Ideally, the neg should print without any effort } on grade 3 paper. I only use Ilford archival matte fiber paper, so } extensions to RC papers may or may not directly apply. I also } do not use variable contrast paper. But the principles are extensible } I would think. } } Being all-digital with my SEM, I use realtime histogram feedback } to adjust gain and dynamic range. For TEM, I have no direct } experience. But I might suggest that the same photo techniques } for fine art neg work may apply to TEM negs. If you think that this } might be true, read on. Otherwise, nevermind. } } } A neg with bad dynamic range (huge extremes of EV or low contrast) is not } going to produce a very good scan. This is the realm of drum } scanners with 4.0-6.0D. Pixel resolution is subordinate to } D range in this case. If one is going to print a neg on paper, } that is one aspect of the problem. If the neg is to be output } to a magazine or printed page, that is another aspect. So the } crux of the matter is what the actual intended end use of the } neg is to be? If it is a nice print, OK. If it is a magazine or litho } output, these are 133lpi. Doing scans at 4000dpi only to } print them in a magazine at 133lpi is rather silly. One would } probably be better off just printing the neg and scanning at } 300dpi. } } But the 300 dpi is the effective dpi for the size of } the final image....not the original neg. So, if the neg is say } 3" x 4" and is to be printed at that same size, a 300 dpi scanner } should do the job (ignoring tonal range for the time being). } If the neg is to be printed at twice its physical size, then the } scanner has to have twice the resolution (600 dpi). And it } goes on from there. Thus, for most image output methods } on a printed page, 600-1200 optical dpi should be plenty of } resolution. If you agree with this discussion at this point, } then the resolution factor should no longer be an issue. } What remains is D range. } } (Note that commercial images are created at high resolution } to accommodate manipulation and merging with other images. } This way, image quality can gradually be reduced without } affecting the final result.) } } But the negative is the key item in all of this discussion. A } bad neg is not likely to do anyone much good. So the challenge } is to get a good neg at the get go. Here we go, back to the } zone system. The idea is to use the neg as a non-linear } image capture media for data which may span a wide } dynamic range. And in so doing, be able to output (print } or scan) this information with fidelity and minimal effort. } This drives D value. } } An unexposed, developed neg will have a transmission of 100%. } It is actually slightly lower due to absorption by the medium, } emulsion and residual anti-dispersion coating. But at 100% } transmission, the neg would have an opacity of 100%input/100%output } or 1.0. Since D=log1/T, the D value for the unexposed neg } is log 1/1=0.0. This then is Dmin. The area with the most } exposure (darkest region) will pass the least amount of } transmitted light. If, for example, this region passes 1% of } the transmitted light, this is a Tval=100%/1%=100. } And D=log (1/100)=2.0. This then is Dmax. The D range } of the negative is Dmax-Dmin=2.0-0.0=2.0. So a scanner } with a D value of } 2.0 would capture the tonal range of this } neg. The tonal range of this neg is 100 tonal variations. } Prints generally have D values between 1.7 and 2.0. } So the same scanner would handle the neg and a typical } print. } } Let's say that the darkest area of the neg transmits 0.1% } of the transmitted source light. This then yields Tval=100/0.1 } =1000. And D=log 1000 = 3.0. Thus, this neg has 1,000 } tonal variations in it from clear to black. If the neg has double } the number of tonal variations (2,000) that would mean } that the opacity is 2,000 and D=log 2000 = 3.3. At 4000 } tonal range, D=3.6. Thus, each 0.3D equates to doubling } the opacity range or halving or doubling the transmission } value. Thus, if a scanner has a D rating of 3.0 (1000 tonal } ranges) and the neg passes a corresponding 0.1% of the } transmitted light, and another scanner has a D rating } of 3.3 (2000 tonal ranges) and the neg passes 0.05% of } the transmitted light--can you really tell one from the other? } } The eye is more responsive to subtle changes in tonal } variations in the white region of an image than in the } dark ones. Thus, it is important to retain as much variability } and rendition in the darkest areas of the negs (the highlights) } but to do so without sacrificing detail in the shadows (clearer } areas of the neg). This is done using the zone system } and expansion and contraction of tonal range. } } The response of the neg emulsion's exposure to light is not linear } over the range of clear to full black. Its response curve is somewhat } like a flattened S. The low exposure region is called the toe, which } consists of the film base + fog density (Dmin). Moving up the curve } is the straight line section (not exactly a straight line), and then to } the shoulder. At this point, increasing amounts of light do not } have corresponding quantitative amounts of change in density. } Further exposure reaches saturation, or Dmax. At this point, } no increase in exposure will change the density of the neg. } } The ultimate goal is to maximize the straight line section and } move Dmax as high as possible, without detrimentally affecting } Dmin. I use contraction to accomplish this. The approach is } to overexpose and underdevelop. With a neg, the rule of thumb } is to expose for the shadows. If there is insufficient exposure } in the shadow areas, there will not be any detail rendered. } } Using Ilford FP4+ film, I rate it at ISO 80 (mfg=125). Development } is done using stock developer diluted 1:1. It is used one time } and discarded. Never replenish or try to refresh the developer. } Development time will vary, depending on the tonal range of the } scene. Here are some extreme examples of how this system works: } (if you are offended by nude images, use the still life links. These } following links are to fine art nudes and still life which were done using the } zone system previously described) } } [nudes] } A. This shot illustrates a typical impossible shot. Inside the room, } the exposure was about EV 3, the outside was dense fog with } a starch white picket fence at EV 11. A scene like this with } eight stops of variation would typically be shot to either render } detail in the highlights (fog and fence) or in the inside room's } details (shadows). To render shadow detail and retain highlight } detail, the zone system was used as described. The image } via this link is as-scanned on a UMAX Powerlook III of a } 6x6cm negative using the transparency adapter. You can } see the detail in the paint on the wall and can see the fence in } the fog. } http://www.photoweb.net/pw_gal_nude/pw_gal_nude_1/g_2.html } } B. This shot shows great shadow detail despite the outside light } creating a hot spot on the model's head. And the window } on the left was rendered, despite the high level of light. } http://www.photoweb.net/pw_gal_nude/pw_gal_nude_1/g_6.html } } C. This shot shows a rather low contrast scene. There are no } remarkable highlights. There is much material in shadow. } Overexposure and ensuring at least two zones of exposure } for the shadows and then overdeveloping N+1 achieved } a perfectly printable neg. Again, this neg is shown as-scanned. } http://www.photoweb.net/pw_gal_nude/pw_gal_nude_6/g_3.html } } } } [Still life] } A. high side lighting. } http://www.photoweb.net/pw_gal_still/pw_gal_still_4/g_5.html } } B. Low shadow lighting, highlights present. } http://www.photoweb.net/pw_gal_still/pw_gal_still_4/g_6.html } } C. Even lighting. There is actually more detail revealed } than is showed in this web pix. } http://www.photoweb.net/pw_gal_still/pw_gal_still_3/g_6.html } } } } Non-web images of these negs are of course much better than } those on-line. But I hope these illustrate the points of the zone } system. I do think it can apply to TEM negs. The other point } is to keep in mind the relationship of D values of scanners to } what you are actually going to be scanning. If a scanner has } sufficient resolution, and say a D rating of 3.2. Are you } really going to be able to tell any difference using a scanner } with a 3.4 or 3.5D at much higher cost? If you have a } densitometer, check the D range of some of your negs. They } are probably all less than 3.0. Maybe I'm wrong in this } respect since I have little experience with TEM media. } But the idea is go get the equipment you need for the job } you need (the output destination and the use of the image) } based on the actual media being scanned. Otherwise, there } is a great opportunity to buy capability which will never be } utilized. } } Despite all the discussion of processing negs, as more } digital capture and image processing products come out, } things will change. There are rather simple ways to expand } the contrast of an otherwise low contrast, poor neg. Likewise, } there are ways to extract subtle detail from negs which have } blown out highlights. More about this later. } } gary g. } } } Reference: } Adams, A. (1981). The negative. Boston: Little, Brown and Company. } ISBN 0-8212-1131-5 (twelfth printing, 1992). } } } } At 01:07 AM 4/12/2001, you wrote: } } } Usualy to reduce contrast you under expose the film. Then develop it to } } the disired density using a developer that generates low contrast. One way } } to reduce contrast is to dilute you developer by a factor of 2, 4 or more } } with water. It extends the developing time a good deal but it reduces the } } contrast. You might also look at low contrast developers that work with } } the film you are using. } } } } A few question on rec.photo.darkroom will get you more information than } } you can handle and some of it will actually work. } } } } Gordon } } Gordon Couger gcouger-at-couger.com } } Stillwater, OK www.couger.com/gcouger
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I tried sending this earlier, but seems to have not made it.
The 840 has some pots (potentiometers) on one of the circuit boards that are used to adjust out any image shift during stigmation adjustments. They probably adjust some bias voltages, or some offsets, in the stigmator circuits. They can be adjusted so there is no image shift.
If the pots can't eliminate the image shift, then a component has probably gone bad, rather than just drifted a bit. If you have the manual with the schematics, they should be identified in there. If you don't have the manual, I might be able to find them in ours. Let me know.
The 18th Annual NESM Woods Hole Symposium will be held May 11-12th, at the Marine Biological Lab at Woods Hole, MA.
This meeting is supported by members of the Connecticut Microscopy Society (CMS), the Metropolitan Microscopy Society (MMS), and the New York Society of Experimental Microscopists (NYSEM).
Pre-registration is encouraged and is a must if you plan to attend the Friday night dinner. Inquiries re: registration for this meeting should be directed to Mary McCann (617) 484-7865 or by email: mccanns-at-tiacc.net. Advance regis- tration including dinner on May 11th is $40.00 for NESM members, and $55.00 for non-members (this cost includes a one- year membership to NESM).
PLEASE NOTE: ADVANCE REGISTRATION MUST BE RECEIVED no later than MAY 4th!!! Registrations received after May 4th will NOT include dinner.
Friday, May 11th begins at Noon and consists of 2 sessions with an afternoon coffee break. Following the presentations, a cocktail hours and dinner will commence in the Swope Center.
Saturday, May 12th, NESM will present a symposium on Remote Access Microscopy. Afterwards, there will be commercial exhibits and posters on display in the Swope Center. Presentation of Poster and Photos-As-Art Awards and Door Prizes will follow. This year lunch at the Swope Center will be optional; those interested will pay $16.00. After lunch, two 45-minute tours of the Marine Resource Center will take place.
NESM welcomes new members to the Society! Please join us for this most enjoyable meeting.
Peggy Sherwood, Corresponding Secretary NESM -- Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) sherwood-at-helix.mgh.harvard.edu
You have a really good point for discussion about actual available exposure time. Real world nudes and still lifes can be for several seconds exposure. What is it for a typical TEM neg? I don't know. I suspect the same drift problems occur in TEM as they do in SEM. So I too like to get the shot as quickly as possible yet with minimum noise. So it is a tradeoff of pixel density and pixel dwell time. Some original loss can be made up later using the computer. With a digital active scan and image capture system synchronized to 60 Hertz, a SEM shot at 3000x3200 pixels takes 96 seconds at 10uS dwell time. This works fine. But for a TEM which may not have sync to line frequency, I can see that image shift is a big problem. Are TEM cameras and scanning routinely synchronized to line frequency?
I used Diafine and Accufine back in photojournalism days to push process TriX to ISO1600 or beyond. The job was to shoot basketball and football games without strobe. The reason was to gain even lighting and high contrast shots. Since the pix were physically small, the higher grain was not a big issue.
Your Diafine approach to TEM negs sounds like a solid method. Indeed, the key is to get the shot and make a print with minimum amount of tweaking (burning, dodging, etc.). If one were to only have to make one print ever from one neg, it would not matter all that much. But if more than one print is or will be made, reproducibility is a tough issue with a less than perfect neg.
I see a similarity in your approach and what I was talking about. Your procedure limits the development time of the neg. But while it limits the time for dense areas, it limits the time for the whole neg too. The procedure I described ensures that as much light information is captured on the film, but extends the tonal range of the film by reducing the strength of the developer and the development time.
I already have de-rated the speed of the film in this procedure. It may be that doing this with TEM negs makes the resulting absolute speed too slow. What is the rated ISO speed of some TEM media in use today?
gary g.
At 01:56 PM 4/13/2001, you wrote: } Reply to: RE: Film Processing & dynamic range & scanners (longish) } I saved your extensive info about negs. I would not use Diafine for } regular scenic photos--too low in contrast. However, the manner in which } split developers achieve this is to LIMIT the amount of developer } available to "dense" negative areas. I only use it for contrasty TEM } negs. that I routinely print on #2 or #3 paper , whichever is more } pleasing. By having easily retrievable info in all of a negative, less } time should be needed by any final printing method. The developer has a } long tank life and works at room temperature. An extreme test of TEM negs } made at 4 f/stops equivalent underexposure still printed nicely on #3 } grade paper. } Of course I used a very long 12 mimutes in each half of the split } developer, but this can allow short exposure on specimens with a } "drifting" problem and save an expensive experiment. } As primarily an old "wet" printer, I enjoyed your thorough } explaination and have heard of the zone system and the great A. Adams. I } like to get good results with little effort, thats all. If you ever want } a copy of the Diafine Co.'s explaination of it's product, I can send you } one. Nearly all of my published photos in Ultramicroscopy were done with } this stuff, including a "lucky" cover photo on Ultra. 25 (1988) 351-354. } } Bernie Kestel E-mail: {kestel-at-anl.gov} } Materials Science Division } Argonne National Lab } 9700 So. Cass Ave., } Argonne Il., 60439 } Gary Gaugler wrote:
Has anyone had experience in determining the concentration of particles in a solution by counting on EM grids in a similar way to using a haemocytometer? Is it possible to count the particles in a defined number of grid squares then calculate back to the area of the grid and the amount of solution which was applied and allowed to dry down?
Dear Richard, I can forsee one difficulty. The particles may not be deposited uniformly due to a number of factors: 1) Evaporation of the applied drop of specimen will not be uniform, so the particles could be dragged in toward the grid center as the edges evaporate (or they could be preferrentially deposited toward the edges if they are hydrophobic). 2) The grid surface may not be flat, causing particles to deposit preferrentially in the centers of the grid squares--if these are lower--or near the grid bars. 3) The particles could be preferrentially deposited either on top of the grid bars or on the open areas, which could look like uniform deposition, but would give erronious quantitation. Of course, one could test this by depositing a known amount of suspension of a known concentration of the kind of particles to be measured, then seeing if the grid was covered uniformly and if the calculation gave the correct result. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
I asked if xylene was the best solvent for cleaning immersion oil off a non-immersion objective. I recalled in my brief training that xylene was safe but alcohols dissolve the more common lens adhesives (I could be wrong and have this reversed). Here is the array of responses I received. The diversity may interest you. THANKS to all for your advice.
1 We regularly use xylene with a cotton tipped applicator to clean our lenses and remove immersion oil. This was recommended to us by the supplier of our lenses and microscopes for a number of reasons. The primary reason is that xylene effectively 'cuts' the oil, without damaging the lens. Acetone will affect the lens coating, and Isopropyl alcohol only rinses the oil without effectively removing it. Hope this helps
2 You are lucky that the lens elements did not dislodge. Use alcohol on lens paper. Never use xylene or toluene; these solvents can dissolve the cement used to seat the various lens elements.
3 I would recommend Sparkle Glass Cleaner. I only use solvents as a very last resort (unless you really, really do want to upgrade) The cements are soluble in most solvents. With intractable grime, I use a cotton tipped applicator moistened with either xylenes or toluene and shake all excess off. The tip must be moist, not wet or dry and make single passes until the grime come off. Only enough solvent on the applicator to dampen the surface, not enough to wet.
4 For day to day cleaning of lenses, including oil on the 40X. (I can't believe this is the first time! The graduate students and Post Doc's drag the40X through the oil at least twice a week. I have given them repeated instructions but they seem intractable) Invert an ocular and examine the surface for cleanliness. If it is clean, don't clean it. Dust off any loose debris. Check the lens again. Moisten a cotton tipped applicator in Sparkle and wipe the lens 1 swipe with a rolling action to present a new surface and lift off any grit. Discard. Take a dry applicator and remove the film. Check the lens with an inverted ocular to see if it is clean. Do no more than is necessary. There has been an ongoing rampage on the list about lens cleaning.... to solvent, not to solvent... lens tissue, not to lens tissue etc.. I can append you a couple, and a microscope maintenance handout if you would like. It will be a bit long about 15 pages (38K).
5 The care instructions for my immersion oil suggest cleaning with a soft cloth or lens tissue (no Kimwipes) moistened with ether/alcohol (7:3) or xylenes. My microscope manuals recommend removing finger prints using alcohol. Sounds like you're good to go.
6 We use 70 % isopropanol to clean emersion oil off of our lenses.
7 Cleaning with xylene is a little drastic. If the lens eventually gets xylene in behind the cement it could do some damage and you would have to send it in to Nikon for repair. I use Kodak lens cleaner and some lens cleaning tissues (not regular Kim wipes) to get the oil off, constantly checking them under a dissecting scope to make sure that it is all removed. It works well on some expensive lens that we have here.
8 We use Green Soap from the pharmacy. It works the best with ultra pure water.
9 Actually, you've got your solvents reversed; alcohol is usually safer. But safer still is diluted detergent "Joy" or similar, followed by water. Use alcohol only if that doesn't work. And fumes from xylene, toluene, etc. are best avoided.
10 Back in the pre PC (chemical) days, toluene was a recommended } solvent for cleaning lenses of immersion oil. Now, we at National Steel, } wipe off excess oil with lens tissue and then clean the lens with Kodak lens } cleaner solution.
Ritchie, What you are describing is a stigmator drive that is out of calibration. If you look on the schematic for the stigmator drive, you should see the X(or Y) control that goes to an op-amp that in turn feeds a voltage divider which in turn controls a different coil in the stigmator coil assembly. Each part of this voltage divider has its own trim pot for calibration purposes. Simply rock the X stigmator control back and forth while adjusting each pot for minimum image shift. Then, increase your mag & repeat. You should be able to get all of the shift completely out. Good luck!
Gary M. Easton, Pres. Scanners Corporation SEM/EDS/IMAGING Sales & Service
----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, April 13, 2001 2:12 PM
Thanks very much to all those who responded to my post.
My hands didn't bleed at all.
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
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At 12:53 PM -0400 4/10/01, Ronald Anderson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am sending this message again because it was blocked by a list server filter - presumably because 'home' is part of your e-mail address.
Malcolm
-------- Original Message --------
Please reply directly to:
} From: {hmskaug-at-tartarus.uwa.edu.au} } } Subject: TEM } } } Hi, } } I am doing some preperation work for an assignment, and have a question. } } If a fresh unfixed piece of brain tissue (weighing approx. 1 gram) is } received in a diagnostic electron microscopy laboratory, how would I } proceed with the specimen? The brain is taken in surgery within the last 5 } min. } } If I was asked to carry out a rapid viral diagnosis on the tissue, what } precautions should I take, and how would I proceed? } I read that prions are able to survive fixation. What then? } } A respond is very much appreciated. } } Thank You ! } } Sincerely, } } Hege Skaug
Greg Erdos Assistant Director Biotechnology Program Ph. 352-392-1295 University of Florida Fax 352-846-0251 PO Box 118525 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
To all colleagues who supply standards for EDS work I need a standard sample of known thickness to check the accuracy of my TEM/EDS system during standardless quant. I need to include light elements (Z {11) as I have a Moxtek SUTW window on my detector. I guess an amorphous glass sample that contains both nitrogen and oxygen with with a very thin layer of carbon on it to prevent charging would do the trick. Can anybody out there supply one? Thanks.
Alan Fox
Professor Alan G. Fox BSc PhD CEng FIM Director, Center for Materials Science and Engineering Naval Postgraduate School Monterey California 93943 USA
This is an apology written in embarassment for some of my private comments about EDX systems that recently got broadcast to the list. I had too-quickly responded offline to a posting and was chagrined to find my comments passed along without the balanced context I would have put them in if I'd been more sensitive to the probability of dissemination. It's too late to rephrase them but I want to emphasize that my recent demos with Oxford, Noran, PGT and EDAX were all highly positive experiences and I came away believing that all are solid and supportive companies offering world class instrumentation. Any system will have many pros and a few cons; our final choice will rest simply on which has a bigger cluster of the former than the latter for our particular set of applications. The cons I had mentioned were subjectively-perceived blips within the significent strengths that each system offers. While cons can serve to pare down the choice, once it's made, as I've discovered through discussion with reference users representing all four companies, users across the board (at least all those I've spoken to) are happy with whichever system they purchased.
Dee
*************************************************************** Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 914/365-8640 F: 914/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
There is a decent test sample out there for checking and monitoring your EDS system called the NiOx sample. One place that you can find it is at Ted Pella. Here is the web site for that product. It has further information on it. http://www.tedpella.com/calibrat_html/TEM7.htm
I am not sure what you mean by the accuracy of your EDS system. You will need to calibrate it and determine whether you are having problems with icing. You can do that with the NiOx sample. The literature that comes with the sample or that you can get from Ted Pella will tell you how to do this. It is also tells you how to monitor your system's performance over time.
If you want to check for N2, just get some Hexagonal BN, crush it between two glass slides, and collect it on a carbon coated grid. You will easily find thin particles.
You hit on a couple of problems with your request. You will have a difficult time ascertaining the thickness of any sample accurately, but with glass, you will only be limited to doing it with EELS or contamination spots. With glass, the composition can change under the beam as elements particularly alkali elements diffuse out of the irradiated area. The composition of glasses can change depending on the depth from the surface and so where you are can make a big difference. I do not know where you would get a glass with a N2 concentration. You can also cause the glass to soften in the beam in very thin areas if care is not taken. Higher accelerating voltages help here tremendously.
I have found that frequently I do not need to coat my glass samples in a 200 keV TEM. I did have to do it when I used a 100 keV machine. For cross section samples, I started using Si blanks as half of the sample and it seems to have eliminated heating and charging problems.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
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} -----Original Message----- } From: Alan Fox [mailto:fox-at-nps.navy.mil] } Sent: Monday, April 16, 2001 12:50 PM } To: MS listserver } Subject: Standard sample for EDS quant } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------- } ---------. } } } To all colleagues who supply standards for EDS work } I need a standard sample of known thickness to check the } accuracy of my TEM/EDS system during standardless quant. I need to } include light elements (Z {11) as I have a Moxtek SUTW window on my } detector. I guess an amorphous glass sample that contains } both nitrogen } and oxygen with with a very thin layer of carbon on it to prevent } charging would do the trick. Can anybody out there supply one? Thanks. } } Alan Fox } } } Professor Alan G. Fox BSc PhD CEng FIM } Director, Center for Materials Science and Engineering } Naval Postgraduate School } Monterey } California 93943 } USA } } Tel (831) 656 2142 (work) } (831) 657 9239 (home) } Fax (831) 656 2238 } } }
May I suggest that we consider some ground rules regarding postings of summaries of replies to questions? It happens on occasion that replies which are assumed to be made in private get posted publicly, sometimes to the embarrassment of those doing the replying. This is, I'm sure, never done with any bad intent and is usually harmless , but can nonetheless be a little disconcerting.
I have posted summaries myself without thinking of the possible consequences, so I'm not pointing any fingers. It's something that's easy to forget about until you get caught yourself.
I would suggest as starting points that: 1) questioners always make clear their intent to post a summary, 2) that the identities of the repliers be removed from summaries (this is often done anyway), and 3) that all who reply to a question indicate if they want their replies to be kept private.
Anyone else have any thoughts on this matter?
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
{FONT face=3D"MS Sans Serif"} {FONT size=3D2} =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D {BR} {FONT color=3D"#FF0000"} {B} Removal Instructions: {BR} {/B} {/FONT} To be removed from our "in house" mailing list {BR} {a href=3D"mailto:remove.promo-at-yourfreehosting.net?subject=3Dremovepromo"} = click here {/a} {BR} and you will automatically be removed from future mailings. {BR} =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D {BR} {FONT color=3D"#0000FF"} {B} Currency Trading Made Simple! {BR} {/B} {/FONT} {BR} {FONT color=3D"#008000"} {B} Do You Have The Yen To Be a A Millionaire? {BR} {/B} {/FONT} {BR} 200% return in less than 90 days! {BR} {BR} Unique Strategy Trading in the International Currency Markets! {BR} {BR} Largest MarketPlace in the World! {BR} {BR} Get our Reports, Charts and Strategies on the U.S. Dollar vs {BR} Japanese yen and euro dollar. {BR} {BR} {FONT color=3D"#0000FF"} {B} Example: {BR} {/B} {/FONT} {BR} A $5,000 Investment in the yen vs the dollar, "properly positioned", {BR} on 08/18 could have returned $15,000 on 09/19/99. For your FREE {BR} information package, contact us today. {BR} {BR} {a href=3D"http://www.independent-hosting.com/FreeHosting/290357/intro.asp= "} click here {/a} {BR} {BR} {BR} {BR} =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D {BR} {FONT color=3D"#FF0000"} {B} Removal Instructions: {BR} {/B} {/FONT} To be removed from our "in house" mailing list {BR} {a href=3D"mailto:remove.promo-at-yourfreehosting.net?subject=3Dremovepromo"} = click here {/a} {BR} and you will automatically be removed from future mailings. {BR} {BR} You have received this email by either requesting more information {BR} on one of our opportunities or someone may have used your email address. {= BR} If you received this email in error, please accept our apologies. {BR} (Any attempts to disrupt the removal email address etc., will not allow us= to {BR} be able to retrieve and process the remove requests.) {BR} =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D {BR} {BR} {BR} {/FONT} {/FONT} {/BODY} {/HTML}
I would need to prepare DNA for TEM using spreading/shadowing in Cytochrome C films. My pilot experiments using just plasmid DNA and the Lang & Mitani method (Biopolymers, 9, p.373, 1970) worked rather poorly and I would like to hear from people with some experience in this method. What are the critical points here? Purity of water/chemicals, Cyt C concentration, time? Any hints would be appreciated.
Do I understand you believe this is CJD or prion infected tissue? If so several methods of disinfection have been suggested (using the word disinfect loosely). No one is sure exactly what will inactivate this prion and the procedure has been disgusted on Histonet for routine pathology giving the latest suggested methods. I can attempt to forward some of that information to you along with the website address. Let me know if you would like the information. Most of us are alittle frightened of this one as it can take years to manifest. Pamela A. Marcum Histology/Microscopy Product Development Manager 400 Valley Road Warrington, PA 18976 Phone: 800-523-2575 Ext 167 215-343-6484 Ext 167 Fax: 215-343-0214 E-mail: pmarcum-at-polysciences.com
-----Original Message----- } From: Greg Erdos [mailto:gwe-at-biotech.ufl.edu] Sent: Monday, April 16, 2001 9:17 AM To: Microscopy-at-sparc5.microscopy.com
Please reply directly to:
} From: {hmskaug-at-tartarus.uwa.edu.au} } } Subject: TEM } } } Hi, } } I am doing some preperation work for an assignment, and have a question. } } If a fresh unfixed piece of brain tissue (weighing approx. 1 gram) is } received in a diagnostic electron microscopy laboratory, how would I } proceed with the specimen? The brain is taken in surgery within the last 5 } min. } } If I was asked to carry out a rapid viral diagnosis on the tissue, what } precautions should I take, and how would I proceed? } I read that prions are able to survive fixation. What then? } } A respond is very much appreciated. } } Thank You ! } } Sincerely, } } Hege Skaug
Greg Erdos Assistant Director Biotechnology Program Ph. 352-392-1295 University of Florida Fax 352-846-0251 PO Box 118525 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
To all colleagues who supply standards for EDS work I need a standard sample of known thickness to check the accuracy of my TEM/EDS system during standardless quant. I need to include light elements (Z {11) as I have a Moxtek SUTW window on my detector. I guess an amorphous glass sample that contains both nitrogen and oxygen with with a very thin layer of carbon on it to prevent charging would do the trick. Can anybody out there supply one? Thanks.
Alan Fox
Dear Alan, I can't get you a standard with all the qualifications you want, but I do have some standards prepared by Chuck Fiori. The matrix is a lithium borate glass, and there are several blocks with various elements evenly dispersed in the matrix. The down side is that you will have to melt the glass, prepare thin specimens--by blowing with a platinum straw--break off pieces from the bubble, and put them on (or in) a suitable grid. I have found that folding grids work well. Of course, you will have to measure the thickness after you prepare the specimen. Anyway, it is amorphous glass with light elements, including oxygen. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
We are trying out the suggestion of using Nile Red to increase fluorescence. This seems very promising at this stage.
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz
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We are looking at purchasing a stereo microscope with fluorescence capabilities primarily to look at GFP fluorescence in plant specimens. Does anyone have any particular comments on the relative merits of the systems produced by the major microscope manufactureres (Leica, Nikon, Olympus and Zeiss). We also have a dichotomy amongst users on whether to provide film or digital cameras for recording - my own preference is more on the digital side but again has anyone any comments or suggestions.
Thanks
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz