Because the lamp in mounted in this microscope with the socket up, large amounts of heat are concentrated in the lampsocket. You are also drawing a lot of current through the connections. Check the cables that connect the power supply to the lamp, there are two, a long one and a short one. You are looking for damage to the connectors, four of them. Changing these cables or assuring that the contacts are clean and tight may solve the problem.
Don't know if anyone makes something commercially, but a lamp current control based on feedback from sampling the light intensity should do it.
Woody
} -----Original Message----- } From: Doug Cromey [mailto:Cromey-at-Arizona.edu] } Sent: Friday, February 01, 2002 4:04 PM } To: microscopy-at-sparc5.microscopy.com } Subject: LM: lamp stability problems in time-lapse } } } -------------------------------------------------------------- } ---------- } 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 } -------------------------------------------------------------- } ---------. } } } Listers, } } We have an Olympus IMT-2 set up for time lapse imaging using } DIC. During } our over-night exposures we find that the lamp does not } provide consistent } illumination, dimming occasionally. This makes for somewhat } less than } satisfactory movies. Is there a way we could make the lamp } more stable? } } Details: 50W tungsten bulb, electrical power goes through a } UPS before it } enters the microscope, the lamp is typically at a setting of } about 10/12 on } the LCD, I will concede that the UPS is old & I'm unsure of } the state of } the batteries, although I'm inclined more to believe its the } lamp or lamp } housing. } } Thanks for your comments. } } Doug } } .................................................................... } : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : } : Research Specialist, Principal University of Arizona : } : (office: AHSC 4212A) P.O. Box 245044 : } : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : } : (FAX: 520-626-2097) (email: Cromey-at-Arizona.edu) : } :...................................................................: } http://swehsc.pharmacy.arizona.edu/exppath/ } Home of: "Microscopy and Imaging Resources on the WWW" } }
I agree with Gary Gaugler in general but, if you only need to power a 50w light source, a good, adjustable, regulated power supply for this alone (12 volts, 5 amps???) can be obtained for much less than $500. Just run the lamp directly from this.
Jim P.
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-- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU 3D Microscopy of Living Cells Course, June 10 - 20, 2002, UBC, Vancouver Canada Info: http://www.3dcourse.ubc.ca/ Applications due by March 15, 2002
There has been a recent thread discussing methods for using Photoshop to place micron markers on micrographs. One thing that people seemed to want was a method to directly use the (presumably known) image magnification to create the final result. In response, I've created a plug-in for Photoshop (and compatible programs) that does this. You enter the magnification of the image (e.g., 1000x), the dpi with which it was acquired (e.g., 300 dpi for your scanner, or the corresponding pixel spacing for your camera), and the length of the bar you desire (in microns), and the program draws and labels the bar in the lower right corner of the image using the selected foreground and background colors. Using a Photoshop action, you can easily apply this procedure to an entire folder of images.
This plug-in is freely downloadable for use on either Mac or Windows computers from the ReindeerGraphics web site, at {http://www.reindeergraphics.com/free.html#entermag} . It can be used on 8 and 16 bit grey scale images or 24 or 48 bit RGB color images. While it is compatible with the Fovea Pro and Image Processing Tool Kit software, it does not require them (but please do see what other kinds of processing and measurement are available when you visit the web site).
If you have comments on the plug-in, or urgent needs for other features or capabilities, please let me know.
Very good point, Jim. Yes indeed. For one little lamp, a regulated DC supply will do nicely. I use these myself as well. The only area of concern, or potential difficulty, is mating the lamp itself to the supply. If I recall the IMT-2 correctly, it has a separate lamp house with interconnecting cable to the un-regulated AC supply in the stand. She would either have to find a mating connector to affix to the DC supply or whack off the current connector and do whatever is necessary to mate with the DC supply. Either way, it is definitely cheaper than a dual conversion UPS.
Powerware makes lower VA rating units, down to I think 500 VA. These units are a few hundred dollars. I used to use DC supplies for 'scope lamps but with all of the bad power problems last year here in California, the dual conversion UPS solved many problems. I got way larger units than I probably needed, only to ensure long backup time.
A good source of supply for various DC supplies is http://www.digikey.com
By all means, do not get a B&K 10Amp unit. It has a soft start over-current feature which will trip with a halogen lamp. When cold, they have low resistance and have high starting current. I found out the hard way about the B&K.
gary g.
At 11:09 AM 2/2/2002, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Try http://www.carnoy.org/ - you can download Carnoy program from this web site. Carnoy is $15 shareware. It is written at Katholieke Universiteit Leuven, in Netherlands http://www.kuleuven.ac.be .
First you have to calibrate the software for a particular instrument by measuring known features of images taken with calibration sample at various magnifications. Result of each measurement has to be written into the menu along with the magnification value. Once set, use of the program becomes simple. Just open the image file, then open the menu, choose magnification value, length unit (from kilometer to nanometer- it will also work for maps), number of units (length of the scale bar), and appearance of the scale bar and characters (color, height, and location), and click OK. Program will automatically superimpose scale bar and a legend on the image. All these things can be set as default. Then only 3 clicks will put scale bar on the image (open menu, choose mag., click OK).
You can undo the scale bar before the image file is saved, but once saved, scale bar becomes part of the image.
Other features will allow you to define and count particles, measure perimeters and densities, and convert image files into various formats back and forth.
The only inconvenience which I encountered while using Carnoy software was somewhat different response of the controls, as compared with expected response of a standard Windows program. But technical support via e-mail was adequate.
Disclaimer: SIA does not have any financial interest in Carnoy software. We are just satisfied customers.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
This message is made of 100% recycled electrons. ----- Original Message ----- } From: {"kellymcg-at-seas.upenn.edu"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 31, 2002 2:53 PM
You can get power supplies that have sensing circuits built in them. I would require some interface circuitry to be build to lock the power supply to the light output.
Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
: Don't know if anyone makes something commercially, but a lamp current : control based on feedback from sampling the light intensity should do it. : : Woody : : } -----Original Message----- : } From: Doug Cromey [mailto:Cromey-at-Arizona.edu] : } Sent: Friday, February 01, 2002 4:04 PM : } To: microscopy-at-sparc5.microscopy.com : } Subject: LM: lamp stability problems in time-lapse : } : } : } -------------------------------------------------------------- : } ---------- : } 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 : } -------------------------------------------------------------- : } ---------. : } : } : } Listers, : } : } We have an Olympus IMT-2 set up for time lapse imaging using : } DIC. During : } our over-night exposures we find that the lamp does not : } provide consistent : } illumination, dimming occasionally. This makes for somewhat : } less than : } satisfactory movies. Is there a way we could make the lamp : } more stable? : } : } Details: 50W tungsten bulb, electrical power goes through a : } UPS before it : } enters the microscope, the lamp is typically at a setting of : } about 10/12 on : } the LCD, I will concede that the UPS is old & I'm unsure of : } the state of : } the batteries, although I'm inclined more to believe its the : } lamp or lamp : } housing. : } : } Thanks for your comments. : } : } Doug : } : } .................................................................... : } : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : } : Research Specialist, Principal University of Arizona : : } : (office: AHSC 4212A) P.O. Box 245044 : : } : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : } : (FAX: 520-626-2097) (email: Cromey-at-Arizona.edu) : : } :...................................................................: : } http://swehsc.pharmacy.arizona.edu/exppath/ : } Home of: "Microscopy and Imaging Resources on the WWW" : } : } : :
After doing a bit of searching myself, I posted a request to a mailing list for The Gimp, asking how one might write a script to enter scale bars, using existing images at various magnifications. The Gimp (GNU Image Manipulation Program) is a free software project that works on GNU/Linux or Windoze, I think, as well as Mac OS/X, I think also. It is highly capable.
I wanted to show that there are other ways to do things than the old, hackneyed solutions.
Alan Davis
Begin forwarded message:
We had a similar problem with a metallograph. As it turned out, the dealer included bulbs with oxidized leads. This led to arcing and subsequent damage to the socket. We polished the contacting surfaces in the socket, sputter coated them with gold, returned the old bulbs to the dealer and purchased bulbs from another dealer with newer stock. The illumination is stable and we haven't replaced a bulb in many years.
At 01:09 PM 2/2/2002 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gtg457a-at-prism.gatech.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, February 3, 2002 at 12:54:22 ---------------------------------------------------------------------------
Email: gtg457a-at-prism.gatech.edu Name: Christal Moore
Organization: Georgia Institute of Technology
Education: Undergraduate College
Location: Atlanta, GA USA
Question: My group in a class is working on a project pertaining to electron microscopy. We do know that current imaging methods either have lower spatial resolution or lack the temporal acquisition capability. We are looking to for a new method or a new way of using existing ones that would have spatial resolution sufficient for depicting the smallest possible cellular structures, and temporal resolution suitable for visualizing as many cellular processes as possible. We have just begun researching electron microscopy and do not know that much about it, such as to why are there are obstacles with the spatial resolution and temporal resolution? Do you know of any new methods you could share with the group, or perhaps a good staring point for us to being searching? Also, is it possible to view cellular processes? What are the steps for doing so? Do you suggest any other experts to contact? Thank you for your time.
We are hoping to stain the nuclei of polychaete worm larvae with propidium iodide. The problem is that due to small sample size, it's kind of a "one-shot" deal and I have no protocol to refer to for concentrations of stain to use. I've searched the web and found protocols saying everything between 0.5mg/ml for sea urchin larvae to 1 micromolar for Xenopus embryos. Should we err on the side of excess and go with the really high concentration?
Background prep info: fixed in 4% paraformaldehyde in sea water labeled with anti-tubulin FITC-conj. antibody
The info sheets from Molecular Probes also recommends RNAse pretreatment. Do people have input on this?
Thanks, Pauline Yu Manahan Research Lab http://www.usc.edu/manahanlab
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
After doing a bit of searching myself, I posted a request to a mailing list for The Gimp, asking how one might write a script to enter scale bars, using existing images at various magnifications. The Gimp (GNU Image Manipulation Program) is a free software project that works on GNU/Linux or Windoze, I think, as well as Mac OS/X, I think also. It is highly capable.
I wanted to show that there are other ways to do things than the old, hackneyed solutions.
Alan Davis
Begin forwarded message:
?Xianglin,
Melting temperature of AlSb = 1338K (J. Phys. Chem. Solids 36 (1975) p.931) microhardness of AlSb = 413 kg /mm2 or 359 kg/mm2(From Landolt-Bornstein, vol 17, semiconductors, Springer 1982) Good luck.
Young W. Kim, Ph.D. Research Professor School of Materials Science and Engineering Seoul National University Kwanak-ku Shinlim-dong San 56-1 Seoul, Republic of Korea 151-744
-----Original Message----- } From: Xianglin Li [mailto:Xianglin_Li-at-student.uml.edu] Sent: Monday, February 04, 2002 1:54 AM To: Microscopy-at-sparc5.microscopy.com
Hi, all
I am looking for some parameters of III-V semiconductors. If you can help me, I will very appreciate for that.
1. Melting point of AlSb. 2. Hardness (on the Mohs scale) of GaN, AlSb, and InSb.
Thanks in advance!
Sincerely yours, Xianglin
Center for Advanced Material Department of Chemical Engineering University of Massachusetts, Lowell Xianglin_Li-at-student.uml.edu Tel: 978-934-3411
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pooley-at-tidewater.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, February 3, 2002 at 19:28:15 ---------------------------------------------------------------------------
Email: pooley-at-tidewater.net Name: Alan S. Pooley, PhD
Organization: retired, volunteer at Umaine marine center
Education: Graduate College
Location: Newcastle & Walpole Maine
Question: Is there a good, relatively cheap digitizer (at least 1024 or 2048 square pixels) for the Zeiss DSM940A SEM? Company name or web site info sought please
I was recently sent some images for image analysis. Unfortunately they in .VTI file format. Is there any software which can convert to .BMP or .TIF format?
Rosemary, Sometimes that information can be found in the MSDS (sorry, Material Data Safety Sheet - required in the US) for the chemical. VWR has them on line (probably others do, also).
Ken Converse owner Quality Images third party SEM service Delta, PA
Rosemary White wrote:
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Does anyone have a triocular LOMO head attachment MFN-11 on the microscope? I’m a beginner and I’d like to talk about microphotography concerning: Kind of B/W film Focusing of specimen on the film plane inside the camera Filters Optical microscope setup Exposure film time
I have some qusetions about EM of bacteriophages. They are double-stranded DNA phages for the plant pathogenic bacterium Pseudomonas syringae.
1. Negative staining. Could someone suggest a good stain and protocol for this type of bacteriophage?
2. Embedding in resin. The bacteriophages will be attached to bacteria on an agar plate. Can anyone suggest a good way to handle them. I suspect if I just cut out a piece of agar I could lose a lot of sample during dehydration.
Thanks in advance
Dave
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I do not know any software that can do this, but you could try ctrl+print screen to copy screen and ctrl+insert to paste the file in windows paint program or any other program. After you insert the image crop it and save in BMP format. If you have any problems e-mail me.
Pavel Lozovyy Argo-Tech SEM lab atcsem-at-earthlink.net
} ... } ... I've created a plug-in for Photoshop (and compatible } programs) that does this. You enter the magnification } of the image (e.g., 1000x), the dpi with which it was acquired } ... } } This plug-in is freely downloadable ... at } {http://www.reindeergraphics.com/free.html#entermag} . ...
Thanks John. I wonder however ... what I had noticed fo a variety of image acquisition systems, that a specific constant was needed. That is, even for a specific SEM which indicated the magnification, I had to use a different magnification constant depending on the acquisition software (e.g., JEOL's own or alternative Oxford). How would your plugin address this? (I can tell you one one method, vs the other, acquired only a portion of the other ... e.g., the Oxford acquired a square image from the center of the rectangular JEOL image)
My own method was to create a printable spreadsheet for my SEM users, which (depending on acquisition method) would indicate the "length of a selection box" for a given magnification. This also allowed for user preferred fonts, micron-bar placement, and style (e.g., simple line, outlined box, black on white, W/B, or color). It certainly wasn't automatic, but it was the only way to accommodate users' presentation preferences.
I've dealt with this problem (supply voltage fluctuation of a few volts) by using a constant voltage transformer (SOLA). The problem with the UPS it that it tracks the supply voltage. The only time the UPS' regulated output kicks in is when the supply voltage disappears. My biggest complaint with the Sola transformer is that it is mechanically noisy, so it gets annoying to be in the area for a long time. They also put out a little electrical noise, but if you can physically isolate the transformer and put it on a separate circuit from your analytical instrumentation, then that shouldn't be much of a problem. Most instrumentation has decent filtering on the power line, anyhow.
For critical work, the light output feedback is the best, assuming you can get a sensor mounted somewhere.
Bill William A. Heeschen, Ph.D. The Dow Chemical Company Microscopy, Digital Imaging 1897 Bldg, E-84 / 2040 Bldg, 1330 waheeschen-at-dow.com voice: 989-636-4005 fax: 989-638-6443
-----Original Message----- } From: Gordon Couger [mailto:gcouger-at-couger.com] Sent: Sunday, February 03, 2002 1:30 AM To: microscopy-at-sparc5.microscopy.com
You can get power supplies that have sensing circuits built in them. I would require some interface circuitry to be build to lock the power supply to the light output.
Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
: Don't know if anyone makes something commercially, but a lamp current : control based on feedback from sampling the light intensity should do it. : : Woody : : } -----Original Message----- : } From: Doug Cromey [mailto:Cromey-at-Arizona.edu] : } Sent: Friday, February 01, 2002 4:04 PM : } To: microscopy-at-sparc5.microscopy.com : } Subject: LM: lamp stability problems in time-lapse : } : } : } -------------------------------------------------------------- : } ---------- : } 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 : } -------------------------------------------------------------- : } ---------. : } : } : } Listers, : } : } We have an Olympus IMT-2 set up for time lapse imaging using : } DIC. During : } our over-night exposures we find that the lamp does not : } provide consistent : } illumination, dimming occasionally. This makes for somewhat : } less than : } satisfactory movies. Is there a way we could make the lamp : } more stable? : } : } Details: 50W tungsten bulb, electrical power goes through a : } UPS before it : } enters the microscope, the lamp is typically at a setting of : } about 10/12 on : } the LCD, I will concede that the UPS is old & I'm unsure of : } the state of : } the batteries, although I'm inclined more to believe its the : } lamp or lamp : } housing. : } : } Thanks for your comments. : } : } Doug : } : } .................................................................... : } : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : } : Research Specialist, Principal University of Arizona : : } : (office: AHSC 4212A) P.O. Box 245044 : : } : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : } : (FAX: 520-626-2097) (email: Cromey-at-Arizona.edu) : : } :...................................................................: : } http://swehsc.pharmacy.arizona.edu/exppath/ : } Home of: "Microscopy and Imaging Resources on the WWW" : } : } : :
I have set of images with different magnifications in Photoshop .psd format with additional layers with micron bars on them. Now I can just drag and drop layer with micron bar on new images with the same magnification after I crop them. It works fine for me since number of images in publications and presentations anyway is quite limited. Of course, it is possible to use Photoshop's "automate" to place micron bars on many images with the same magnification.
To create image with micron bar for copying: 1. create new layer 2. draw micron bar with the same length as original bar. 3. using Type Tool type N Microns (or millimeters or whatever else), new (third) layer will be created automatically 4. make background layer invisible 5. click on layer-} merge visible 6. make background visible again 7. save in .psd format with name "Magnification M"
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
I know that this has been asked before but I just want to see if there has been any updated software. I would like to know what is available out there for online scheduling of equipment. Thanks. ______________________________ Roberto Garcia rgarcia-at-unity.ncsu.edu http://spm.aif.ncsu.edu/aif http://spm.aif.ncsu.edu/asm
Dear Christal, First, let me wish your group the best of luck with this project, it is a difficult but worthwhile undertaking. The problem with obtaining spatial and temporal resolution is a complex one. To start I will point out one of the fundamentals of microscopy, the Abbe equation: (I would suggest visiting this web site for a better explanation: http://lsvl.la.asu.edu/bio598L/notes/lightlenses/ ) resolution = (0.612 x l) / (n sin a)
l = (wavelength of light) n = refractive index of medium between the objective lens and the specimen. a = the aperture angle, which is half the angle of the cone of light from the specimen accepted by the front lens objective. This is also called the half aperture angle. (n sin a) is the 'numerical aperture' of a lens, and is usually printed on the side of objectives for light microscopes.
What is important to note is that resolution is a function of essentially two things, your lens and the light you are using. More specifically, the wavelength of the light is related to resolution in such a way that shorter wavelength = better resolution. Electron microscopes achieve their remarkable spatial resolution through the use of electrons as the 'light'. The wavelengths of visible light are in the range of 400-700 nm; electrons have a considerably shorter wavelength, about 0.005nm (recall that matter behaves as a particle and also as a wave). It is because of this shorter wavelength that electron microscopes have such excellent spatial resolution. So, electron microscopes have excellent spatial resolution; why don't they have temporal resolution as well? Temporal resolution is the ability to see change over time. To see a cellular process change over time, you must be looking at a live cell. Unfortunately, it is not possible to do this with an electron microscope for a number of reasons (there are probably others, but these are the most obvious, at least to me). The first has to do with the fact that electrons are very easily scattered when they pass into/through matter; electrons are so easily scattered that the tube of electron microscopes must be pumped down to a vacuum so that the air in the column doesn't scatter the electron beam! If electrons have a hard time traveling through air, one can imagine that the relatively denser matter of a biological specimen would be nearly impenetrable. To make it possible for electrons to get through a specimen, you have to cut the specimen into very thin sections. A typical eukaryotic cell might be on the order of 10-15 microns thick; most sections for transmission electron microscopy are around 0.1 microns thick. After sectioning a specimen, electrons will pass through it. The challenge then becomes being able to distinguish cellular structures. A professor of mine once compared this to looking for chunks of clear Jell-O in a swimming pool. The problem is that electrons (and light in general) pass through most cell components in the same way. To get contrast you have to stain specimens with an electron dense material, usually a heavy metal stain such as Osmium tetroxide (OsO4). So this gives us three major reasons why living cells hate electron microscopy: 1. specimens must be placed in a vacuum while they are being viewed 2. you have to cut the specimen into very thin sections 3. electron dense stains (such as OsO4) are usually highly toxic None of these conditions are compatible with living cells. To make matters even worse, it is necessary to imbed specimens in a plastic resin for them to hold their shape during sectioning (otherwise it's a bit like slicing a tomato with a butter knife). For information on microscopic techniques currently in use I would recommend the following web sites: University of Arizona's web site list http://swehsc.pharmacy.arizona.edu/exppath/micro/index.html Lance Ladic's site on confocal microscopy http://www.cs.ubc.ca/spider/ladic/overview.html You may also want to visit the home pages for the major microscope manufacturers, such as Zeiss, Leica, Olympus, Nikon, BioRad, etc. Again, best of luck to you. -- Russell McConnell Confocal Imaging Facility Technician Department of Neuroscience Tufts University School of Medicine M&V Building room #137 136 Harrison Ave. Boston, MA 02111 Tel. (617) 636-3795
gtg457a-at-prism.gatech.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 } -----------------------------------------------------------------------. } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (gtg457a-at-prism.gatech.edu) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, } February 3, 2002 at 12:54:22 } --------------------------------------------------------------------------- } } Email: gtg457a-at-prism.gatech.edu } Name: Christal Moore } } Organization: Georgia Institute of Technology } } Education: Undergraduate College } } Location: Atlanta, GA USA } } Question: My group in a class is working on a project pertaining to } electron microscopy. We do know that current imaging methods either } have lower spatial resolution or lack the temporal acquisition } capability. We are looking to for a new method or a new way of using } existing ones that would have spatial resolution sufficient for } depicting the smallest possible cellular structures, and temporal } resolution suitable for visualizing as many cellular processes as } possible. We have just begun researching electron microscopy and do } not know that much about it, such as to why are there are obstacles } with the spatial resolution and temporal resolution? Do you know of } any new methods you could share with the group, or perhaps a good } staring point for us to being searching? Also, is it possible to } view cellular processes? What are the steps for doing so? Do you } suggest any other experts to contact? Thank you for your time. } } ---------------------------------------------------------------------------
I operate a JEOL 848 SEM but am limited to less than 30,000x due to at least two frequencies of external noise. The service engineers from JEOL have determined the noise is not due to the SEM. I will be conducting as much of a room survey as possible later in February but I am hoping to obtain some insight into common testing formats, noise sources, and solutions. From what I have been able to learn so far, this noise reduction is something of an art rather than science. Thank you in advance from this microscopist with much to learn.
Sola magnetic resonance transformers are good for this application. However, they are not cheap--especially as their VA rating increases. And yes, they get noisy.
For critical work, I suggest either a regulated DC supply or a dual conversion UPS. The dual conversion UPS never "kicks in" but rather is always producing regulated (frequency and voltage) AC from DC. The DC is either that from rectified line voltage or from the UPS batteries.
gary g.
At 06:43 AM 2/4/2002 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Have any of you compared the Nikon Coolpix 995 (or earlier) with the Olympus C-4040 (or earlier) mounted on a light microscope? They are both comsumer-level digital cameras with about the same technology, and so I'm wondering if there is any reason to choose one over the other. The Olympus has better features on paper, so I'm hoping to hear from someone who uses one. This will be used to supplement the Magnafire SP I'm proposing to buy, since I need a camera that will fit into the eyepiece of our confocal.
Mahalo! Tina
**************************************************************************** * 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* ****************************************************************************
I have been trying to image with negative stain, protein chains and protein dimers with 2.0% Uranyl Acetate. It seems when I pipet onto the grid, various dilutions (10-100x) of the buffered protein dimer sample, I get alot of protein chain formation. I want the dimer form to stay in that configuration-- not link up into chains. I am photographing at 100,000 using 75 kv on a traditional Hitachi 7100 TEM.
Does anyone out there work with these types of specimen? Is there a special way to prep the grid, dry the grid, etc.? Should I use some other type of grid besides the formvar/carbon coated copper grid? Should I sonicate the specimen before I place the sample on the grid?
Thanks for any help you can provide.
Karen Bentley, M.S. (previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
Don't be too quick to accept the service engineers claim that the SEM is not the source of noise in the image. When they can't easily figure out what the problem is, it is too easy for them to blame the "fields," because it is something not well understood by most people. I went through all of the field service people, the service people at the JEOL headquarters in Massachusetts, and an "expert" from Japan. The expert's final conclusion was that the problem was caused by the fields in the room, and not our new SEM. A while later, we moved the SEM some miles down the road to a different building. We had JEOL check the room before moving in, and they blessed the room. When we had the exact same problem, I had a discussion with the people in Massachusetts. Eventually, we had a repaired system that provided excellent images. This all took way too long to resolve. The only company that I am aware of having intimate knowledge of their systems "noise" was Amray. They used a spectrum analyzer on the system, and identified every source of every frequency.
Good luck! Your problem will, most likely, not be this difficult. Darrell
"Curtis Olson" {COlson-at-scpglobal.com} on 02/04/2002 01:12:43 PM
To: {Microscopy-at-sparc5.microscopy.com} cc:
I operate a JEOL 848 SEM but am limited to less than 30,000x due to at least two frequencies of external noise. The service engineers from JEOL have determined the noise is not due to the SEM. I will be conducting as much of a room survey as possible later in February but I am hoping to obtain some insight into common testing formats, noise sources, and solutions. From what I have been able to learn so far, this noise reduction is something of an art rather than science. Thank you in advance from this microscopist with much to learn.
I am a student at Scottsdale Community College. I am working with an ISI SX-40 Scanning Electron Microscope. It has an old polaroid camera that snaps a picture of a CRT image. I am looking for options(and donated equipment) to put these images on to a PC, the big problem is I am on a very tight budget($800) so I am very willing to look for used equipment (card or camera or info) . This machine is old and did not work at all. Over the past 8 months or so I have rebuilt the diffusion pump, roughing pump, new seals, column cleaning, and learned basic operation by myself with this apparatus. And I have learned a lot from following this list server, so I wanted to send out a big thanx to all the listers. Thank you in advance for your time and consideration. Any and all input is appreciated
Wil Kunkel Student Extraordinaire
This email was sent with 100.00% recycled electrons.
I would start with a close inspection of the power supplied to the SEM. Although measuring for stray fields and floor vibration may be in order, a good place to begin is with the A/C power and the potential for ground loops if wired common or shared with other equipment. The ground terminal into the microscope main power supply can often be a good source of noise.
On a positive note, if this noise is a problem as low as 30KX it should be somewhat easier to find.
Regards,
Bob Roberts EM Lab Services, Inc. Tempe, Arizona 85284
Curtis Olson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Announcing the Seventh Annual INTERNATIONAL 11-Day Short Course on
3D Microscopy of Living Cells, June 10 - 20, 2002
Sixth, Post-course Workshop on, 3D Image Processing, June 22 - 24, 2002
Organized by Prof. James Pawley, University of Wisconsin-Madison
in association with the, UBC Brain Research Centre, Prof. Max Cynader, Director. University of British Columbia, Vancouver, BC, Canada
(CORRECTION!! some early brochures were sent out with an incorrect URL. Course info can be found at: ht tp://www.3dcourse.ubc.ca)
THE PURPOSE OF THE COURSE
Modern methods of 3D light microscopy promise a revolutionary improvement in our ability to view living cells. To help convert this promise to reality for a wider selection of biological scientists, the organizers have designed an intensive eleven-day residential course concentrating on all aspects of the 3D Microscopy of Living Cells. Sponsored by the Brain Research Centre at the University of British Columbia, it will be held in June of 2002. The course includes 4 days on 2D techniques, 5 days of 3D techniques and 2 days on 3D measurement and display. It includes everything from basic microscopy to confocal and multiphoton microscopy. A half-day Pre-course is offered for those wishing to brush up on (very!) basic optics.
INTERNATIONAL FACULTY o Stephen Adams University of California-SD o Dan Axelrod University of Michigan o Mark Cannell University of Auckland, NZ o Rainer Duden Cambridge Institute for Medical Research, UK o Ping Chin Cheng SUNY, Buffalo o Stefan Hell Max Planck Institute, Goettingen o Alan Hibbs BioCon, Melbourne, Australia o Iain Johnson Molecular Probes, OR o Larry Keenan Cell Robotics, NM o Ernst Keller Carl Zeiss, Thornwood, NY o Andres Kriete Tissue Informatics, Pittsburgh o Glen MacDonald Virginia Bloedel Hearing Inst, WA o Irina Majoul Max Planck Institute, Goettingen o Felix Margadant University of Sydney, AU o Tim Murphy University of British Columbia o Jim Pawley University of Wisconsin-Madison o Steve Potter California Institute of Technology o Badri Roysam Rensselaer Polytechnic Institute, NY o Lee Tierney Eppendorf Scientific,Albequerque, NM o Michael Weis Agriculture Canada
APPLICATIONS Applicants will submit an application to assess knowledge level and field of interest. Enrollment will be limited to 24 - 32 participants (depending on equipment availability). Selection will be made on the basis of background and perceived need. Those with little previous LM experience will be provided with basic texts to read before the course begins, and should take the Pre-course.
Application forms and other course information from this and past years can be downloaded from the WWW site at
h ttp://www.3dcourse.ubc.ca/home.html
or obtained from:
Prof. James Pawley, Zoology Department., 1117 W. Johnson Drive, Madison, WI. Phone: 608-263-3147 fax. 608-265-5315 Email: jbpawley-at-facstaff.wisc.edu
IMPORTANT DATES Applications must be received by Mar. 15, 2002 Deposit due Apr. 15, 2002 Registration 5:00 - 7:00 pm Sunday, June 9, 2002 Intro. Lecture 7:00 PM, Sunday, June 9, 2002 Last class ends with lunch, Thursday, June 20, 2002 3D IP Workshop Saturday, June 22-24, 2002 -- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jhardy-at-coh.org) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, February 4, 2002 at 15:28:55 ---------------------------------------------------------------------------
Email: jhardy-at-coh.org Name: John Hardy
Organization: City of Hope Medical Center
Education: Graduate College
Location: Duarte, California
Question: Does anyone have experience with, or comments about the "Image Supercharger Upgrade" by Ascend Technical Sales? It would be an add-on image processor for our "mature"(i.e. 1984) Philips 505 SEM. Please contact me off line at: jhardy-at-coh.org Thank you in advance John Hardy City of Hope Medical Center Duarte, CA (626)301-8265
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Email: ever4us-at-comcast.com } Name: Denise Everett } } Organization: Pitman Middle School } } Education: 6-8th Grade Middle School } } Location: Pitman, NJ 08071 } } Question: I've recently become the science coordinator of our school } but my science background is in enzymology so I don't have alot of } experience with microscopy. We are looking to buy some new scopes } and in the process I've been looking at our old ones. The 400x } magnification is pretty unusable on these. Is that supposed to be oil } immersion use only? } These lenses are pretty dirty and have probably not been maintained. } The top objective does not come out for cleaning as far as I can see. } Do I just need to send these to a technician? } } --------------------------------------------------------------------------- Denise -
I'm glad that you've asked for help; we're here to provide it. There are several topics here. First, new microscopes. Let's assume that the scopes that you have are salvageable. Since you're in a middle school, PLEASE consider purchasing "dissecting" rather than compound scopes like the ones that you have. A lot of introductory microscopy for your age group is observstion of thick specimens at lower magnifications; looking at large insects, flowers, shells, etc. So having a mix of types will greatly expand your capabilities. You'll find a detailed discussion of selection criteria on Project MICRO's website (URL below). I suggest 20x monocular dissecting scopes, wich will cost you around $75 each; sources are listed on the MICRO site and you can find an example online at www.microscopeworld.com. Your dirt diagnosis is probably accurate. It would be best if you learn to clean the scopes yourself; you'll then know how to keep them that way. The New York Microscopical Society has members and meeting rooms in New Jersey, and one of their members may be available to show you what to do; I'm copying this Email to Jean Portell {JeanDP-at-aol.com} , one of their officers. I also can provide you with detailed cleaning instructions for teachers, written by a MSA member. 400x is "high dry" - oil immersion is 1000x and inappropriate for middle school. While you're visiting the MICRO website, don't miss MSA's middle school manual, "Microscopic Explorations"; it's an excellent introduction to scientific observation and inquiry, written by the science educators at the Lawrence Hall of Science. If you want a reference book for your own use, here's another listing from the MICRO bibliography:
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; it's unequalled as a basic reference for beginners. Almost half of the book is devoted to simple preparation methods for biological specimens and descriptions (with good illustrations) of commonly encountered organisms. Adult. RECOMMENDED
Caroline
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
I need some help in converting some raw EELS data into formats readable by either EL/P or digital micrograph. The data I have gathered is in .ZAL (Zaluzek) format. The data looks very similar to the .TAD format. Let me know if there is a program that reads .ZAL format and converts it to .TAD or even two column text. Perhaps Nestor would be able to help in this regard, right? :). Thanks in advance,
Prad
Pradyumna Prabhumirashi Dept. of Materials Science Northwestern University Phone: (847)491-7798 Fax : (847)491-7820
Hi, To create an image with a labeled micron bar in 20-30 seconds: 1. Make the info tab totally visible as a separate window on the right side of PS. 2. Crop your image. I assume you are calibrated in cm. 3. Click on the line drawing tool. 4. In the lower left corner, click where you want the marker to start. 5. Drag to the right slightly, then hold down the shift key for a perfect straight line. (Use the option tab to increase the line width (5-15 pixels).) 6. Drag to the right until the D: in the INFO window that shows the length you need on the final printed image You should take into account how you might have changed the DPI values. For example, if you scanned a neg. at 600 DPI and cut it to 200 DPI, then a negative of 10,000X will be about 30,000X on the print and the line length for 1µm should be D: 3.00 CM in INFO. This varies a bit with your printer. Use scanned graph paper to calibrate your DPI to exactly 3x. I use 196 DPI on a HP1200TN to get 3X, after scanning at 600 DPI. 7. CAREFULLY release the mouse button, then the shift key and you have your perfect line. 8. Click on the type tool and click at the position you want the text above the marker. I use centered, 15 point, crisp, black ink, ariel MT, bold, etc. 9. Type in using 15 points the length of the line; 1µm, 0.1µm, 0.3 µms, etc. 10.. Highlight the "1µm" text and copy it as a template to the clipboard for the next image. Click OK. 11. Continue to the next image.
We are in process of buying a TEM for our Science Faculty; it would be used by physicists and materials scientists, and also chemists (inorganic and organic samples).
We have a 300kV FEG machine with ultra hi res polepiece (restricted tilt). We do polymers and inorganics, no problem. We now need a LaB6 machine (budget constraint) with high tilt to be more of a Workhorse Machine.
For the sake of resolution and penetration, we are favoring a 300kV analytical TEM with ~2.1A resolution, and around 40 degrees tilt. This will be perfect for the physical scientists.
HOWEVER: The chemists are worried that they will have trouble looking at their polymer samples in a 300kV LaB6. Can someone help to support me (or correct me) in my thesis that this machine will be able to support them just as well as a 200kV machine? (I know eg that the HT can be reduced to 100 or 200kV, but the chemists are still skeptical - they have no prior TEM experience).
If there is someone with a 300kV LaB6 TEM running polymers, I would be very glad to hear from you! Any references to your papers that would show polymers imaged in a 300kV LaB6 machine would be most welcome (esp any soft copies that I can circulate to our committee).
Thanks to all in advance,
Mark
%%%%%%%%%%%%%%%%%% Mark Yeadon Senior Research Fellow Institute of Materials Research and Engineering 3 Research Link Singapore 117602
Assistant Professor Department of Materials Science National University of Singapore Singapore 119260 http://www.matsci.nus.edu.sg/STAFF/Mark.html
I've been asked if the stage micrometers I use for calibrating our optical scopes are NIST-traceable. Is there such a thing and, if not, what would satisfy requirements for ISO certification in the area of metrology?
Thanks!
Dave Stadden Research Scientist Armstrong World Industries, Inc. Lancaster, PA
Bob Bloodgood, a cell biologist at the University of Virginia, has posted nice images of light microscopes on postage stamps on the web at http://www.med.virginia.edu/med-ed/cell/stamps/index.html
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
Pawley, J.B. Strategy for Locating and Eliminating Sources of Mains Frequency Stray Magnetic Fields. Scanning 7:43-46 (1985).
Pawley, J.B. Use of Pseudo-Stereo Techniques to Detect Stray Field in the SEM. Scanning 9-3:134-136 (1987).
The matter of ground loops, and stray fields from bad house wiring are a bit complex to discuss in emails. Please excuse the self-advertisement.
Jim P.
} Curtis, } } I would start with a close inspection of the power supplied to the } SEM. Although measuring for stray fields and floor } vibration may be in order, a good place to begin is with the A/C } power and the potential for ground loops if wired common } or shared with other equipment. The ground terminal into the } microscope main power supply can often be a good source of noise. } } On a positive note, if this noise is a problem as low as 30KX it } should be somewhat easier to find. } } Regards, } } Bob Roberts } EM Lab Services, Inc. } Tempe, Arizona 85284 } } } Curtis Olson wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU 3D Microscopy of Living Cells Course, June 10 - 20, 2002, UBC, Vancouver Canada Info: ht tp://www.3dcourse.ubc.ca/ Applications due by March 15, 2002
} From the personal responses I've been getting it seems like a lot of people haven't figured out that the Olympus C-3030 and C-4040 cameras can, indeed, be mounted on light microscopes. Olympus can supply all the adapters needed for this task, and can come in with an easier setup, better resolution, and a lower price. The main attraction of the Coolpix seems to be that it can swivel to make viewing the screen easier (I heard a rumor that this feature is about to be discontinued), but either camera would benefit by being plugged into a monitor. I haven't had a chance to compare the cameras, and was hoping to scare up someone who has. Interestingly, the ones who have replied with information have not been from the US, so local marketing must be spotty.
} From the replies so far from people who have tried both, one bought the Nikon, one bought the Olympus.
Aloha, Tina
**************************************************************************** * 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* ****************************************************************************
I'd like to throw out another possible option for creating micron bars on images. The PAX-it Basic Measurement Module (as an add-on for the PAX-it image database product) provides this exact capability. Similar to other software options suggested previously, the Basic Measurement Module also offers other measurement capability such as point-to-point distances, angle measurements, segmented line lengths, parallel line calipers, etc. There is also an Enhanced Measurement Module adding a range of additional features for area detection, blob counting, sizing, and sorting, etc. The measurement modules also provide direct report generating capabilities through (OLE) Automation links to MS Word, Excel, or PowerPoint.
However, what PAX-it and its measurement modules offer, that other options mentioned previously do not appear to offer, is a fully integrated relational database for images and related documents such as Word documents, Excel spreadsheets, PDF files, PowerPoint presentations. Everything relating to a project can be entered into the database as various records with searchable criteria. The database user-interface is an easy-to-understand visual presentation using file cabinets, cabinet drawers, file folders, and thumbnails of the images. And, the entire database can be placed on a file server and, with a special module, made available across internal networks or the internet to be accessed using just a standard web browser interface.
PAX-it is a commercial product and we are a reseller for the product. Hopefully this message is not inappropriate in this forum because we are not the only reseller for the product, and we are not directly soliciting sales of the product through this message. The intent is simply to raise awareness of another option for image measurement and management.
The manufacturer for PAX-it is MIS (847-455-0450) and the web page is www.paxit.com. There are multiple dealers nationwide and interested parties should contact MIS to find the dealer nearest them. (You might indicate that you became aware of the product through the Microscopy ListServer.)
-----Original Message----- } From: Beauregard [mailto:beaurega-at-westol.com] Sent: Monday, February 04, 2002 6:38 PM To: microscopy-at-sparc5.microscopy.com
Hi, To create an image with a labeled micron bar in 20-30 seconds: 1. Make the info tab totally visible as a separate window on the right side of PS. 2. Crop your image. I assume you are calibrated in cm. 3. Click on the line drawing tool. 4. In the lower left corner, click where you want the marker to start. 5. Drag to the right slightly, then hold down the shift key for a perfect straight line. (Use the option tab to increase the line width (5-15 pixels).) 6. Drag to the right until the D: in the INFO window that shows the length you need on the final printed image You should take into account how you might have changed the DPI values. For example, if you scanned a neg. at 600 DPI and cut it to 200 DPI, then a negative of 10,000X will be about 30,000X on the print and the line length for 1µm should be D: 3.00 CM in INFO. This varies a bit with your printer. Use scanned graph paper to calibrate your DPI to exactly 3x. I use 196 DPI on a HP1200TN to get 3X, after scanning at 600 DPI. 7. CAREFULLY release the mouse button, then the shift key and you have your perfect line. 8. Click on the type tool and click at the position you want the text above the marker. I use centered, 15 point, crisp, black ink, ariel MT, bold, etc. 9. Type in using 15 points the length of the line; 1µm, 0.1µm, 0.3 µms, etc. 10.. Highlight the "1µm" text and copy it as a template to the clipboard for the next image. Click OK. 11. Continue to the next image.
I have joint this group recently so, I do not know about the previous discussion. The problem about women radiation workers is that (in my opinion): At birth each female carries a lifetime supply of egg cells. These egg cells are not in their final form but anyway they can be damaged or altered when a female radiation worker is exposed to radiation any time.
To make it short, it is not only what we are doing during pregnancy, it is what we have been doing before pregnancy as well.
Recently, I had a baby and I have cancelled my experimental work at research reactors and my flights during my pregnancy.
Ayten Celik Aktas -at-UIUC
On Thu, 31 Jan 2002, Leona Cohen-Gould 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 Evelyn, } this question came up a while back,so you may want to scan the } archives, but here are my 2 cents... } I have a healthy, happy 9 year old son. I have been doing TEM & SEM } for 25 years. While I was "family pIanning" and pregnant, I wore } double gloves and worked in the hood when appropriate, washed my } hands frequently, followed OSHA guidelines and used common sense. I } still do (except for the double gloves part). Yes, much of what we } use can be dangerous to ourselves as well as our progeny, but with } appropriate care I don't think one needs to take a leave or stop } doing your job. } } JMHO, } Lee }
My requirements are that I have to put a label as well as a scale bar on each optical micrograph and burn CD to archive, all with GLP documentation (we are working towards a 21CFR Part 11 compliant system). I have calibrated our research microscope with various optical systems and photoeyepieces as well as a new macroscope and created about 100 scale bars. We then automated the whole process as much as we could using Actions and Droplets and a Word macro to create a label as well as a list of all the files burned on a CD by filling in a form.
Next we are hoping to be able to write a plug-in that will create a history text file to document each change to a photograph. I asked Adobe if they could do that and the person said that it would be feasible but apparently not enough demand, yet.
I evaluated the Nikon Coolpix 990 and Olympus C-3040 for use on my Olympus SZX12 stereomicroscope when working on-site. When asked, I recommend both cameras.
I chose the C-3040 because Olympus sells a fixed tubelength adapter (C2000Z-ADP) that mounts to the trinocular head of the Olympus stereomicroscope (and my Continuum infrared microscope, which was manufactured using Olympus components). The adapter makes the camera parfocal on Olympus microscopes -- no need to focus an adapter. Also, the C-3040 comes with a remote shutter release as a standard accessory.
Performance on and off the microscopes has been outstanding. In addition to making still micrographs, I use the camera to record digital video of solubility and microchemical tests for clients and colleagues. The live video out function makes focusing and instruction easy (I use a Sony Trinitron 13 monitor on road trips, or an inexpensive monochrome monitor when I fly).
I also used the camera to record my son's first rookie hit in Little League. What does the commercial say? Priceless.
James Martin Orion Analytical, LLC Post Office Box 550 Williamstown, MA 01267 t: 413-458-0233 f. 413-458-5542 www.orionanalytical.com
----- Original Message ----- } From: "Tina Carvalho" {tina-at-pbrc.hawaii.edu} To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, February 05, 2002 1:56 PM
Are you referring to the EMSA format when you are referring to the ZAL format? If you are, there is a program in the EMSA/MAS library that I put in called "EELS_Plot" this last summer. I am about to send Nestor a newer version of this to replace it. It will plot EELS and EDS data in EMSA format and do a few other things as well. If you want to color, display, compare, subtract background, rescale, and display up to five spectra, copy to window applications, and keep notes, this program will do it. If you can't wait, send me your address, and I will send you a copy on CD.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Pradyumna Prabhumirashi [mailto:p-prabhumirashi-at-northwestern.edu] Sent: Monday, February 04, 2002 11:25 PM To: Microscopy Listserver (Microscopy Listserver)
Hello,
I need some help in converting some raw EELS data into formats readable by either EL/P or digital micrograph. The data I have gathered is in .ZAL (Zaluzek) format. The data looks very similar to the .TAD format. Let me know if there is a program that reads .ZAL format and converts it to .TAD or even two column text. Perhaps Nestor would be able to help in this regard, right? :). Thanks in advance,
Prad
Pradyumna Prabhumirashi Dept. of Materials Science Northwestern University Phone: (847)491-7798 Fax : (847)491-7820
We are using a Noran Voyager 3.3 energy dispersive analyzer to generate linescan analyses across catalyst granules. We would like to obtain the data in a spreadsheet format (like Excel or even ASCII) for use outside of the Noran system. We are able to open regular analyses saved in the MSA format but the linescans can not be saved in this format.
Does anyone know how this could be done?
As always, we are very appreciative of any help that you might provide.
JB
-- ############################################################## 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 ##############################################################
In a message dated 2/5/02 5:11:37 PM, neuberger1234-at-attbi.com writes:
} Next we are hoping to be able to write a plug-in that will create a history } text file to document each change to a photograph. I asked Adobe if they } could do that and the person said that it would be feasible but apparently } not enough demand, yet.
The need to document each step in the image processing chain is also important for forensic applications. You should check with Chris Russ at Reindeer Graphics (jcr6-at-AOL.com), who has been working on that need and may have some ideas or even a plug-in for you to use.
What worked for this lab was to ask the female employee in question to inform you when she intended to become pregnant and then to arrange for someone in the lab to take responsibility for sample preparation during the "intention period", 9 months of pregnancy, the maternity leave and the additional months during which the mother was nursing the child. This amounted to a substantial amount of time in our case but I felt better about taking on the added responsibility rather than deal with possibly serious consequences after the fact. Rosemary Walsh
Dr. Russ, We have already contacted Chris and are working on an agreement to help us, or write for us, the plug-in(s). though someone who reports to me is handling this, I will be calling Chris myself to discuss it. It is something that I would like to make available to everybody when done, perhaps through the IPTK next version.
Damian Neuberger P.S. I hope that I can take the class this time around, last time I had to cancel due to travel restrictions.
----- Original Message ----- } From: {DrJohnRuss-at-aol.com} To: {neuberger1234-at-attbi.com} ; {microscopy-at-sparc5.microscopy.com} Sent: Tuesday, February 05, 2002 6:34 PM
To whom it may concern:
I am an undergraduate student enrolled at the Georgia Institute of Technology. I am in a biomedical engineering class, and we have a problem that we must solve involving electron microscopes. I have a few questions that I hope will get answered ASAP:
1. What are the advantages and disadvantages in using electrons for microscopy rather than light? 2. Does the wavelength of the electrons have anythign to do with the spatial resolution that the microscope produces in the final picture? 3. What is temporal resolution and how is it produced in the electron microscope?
Thank you for your time. I greatly appreciate your efforts in helping me understand more of this subject.
Sincerely, Jenny Wang
------------------------------------------------- Sent through Cyberbuzz- A Server for the Students http://cyberbuzz.gatech.edu/
I am looking around for information about geometry based algorithms for analysis of microscopy images. I have some experience with this kind of algorithms, which are extremely powerful but somewhat CPU-hungry and I am looking for improvements.
On the top my personal website you will find examples about what I mean with geometry based analysis of microscopy images of cells and tissues:
} We are using a Noran Voyager 3.3 energy dispersive analyzer to } generate linescan analyses across catalyst granules. We would like to } obtain the data in a spreadsheet format (like Excel or even ASCII) } for use outside of the Noran system. We are able to open regular } analyses saved in the MSA format but the linescans can not be saved } in this format. } } Does anyone know how this could be done?
I have written an application to read the various Voyager file formats (.eds,.lscan,.grey,.xray) on a PC and display/export data in text format. For the linescan format it will display the image file with line and each of the linescans graphs. These can be exported to a text file for use in Excel, etc. I can let you have a copy of this if you wish.
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 1223 334325 Fax: +44 1223 334567 Email: djv23-at-cam.ac.uk
I am looking around for information about geometry based algorithms for analysis of microscopy images. I have some experience with this kind of algorithms, which are extremely powerful but somewhat CPU-hungry and I am looking for improvements.
On the top my personal website you will find examples about what I mean with geometry based analysis of microscopy images of cells and tissues:
I think your instructor's hope would be that you figured out the answers to class problems on your own. Asking an expert in the field and then simply regurgitating that information is a worthless exercise. If you are going to invest the time and money required to earn a degree, you might want to try to learn something along the way. I am a big supporter of listservers but hate to see them used in this way.
} } } To whom it may concern: } } I am an undergraduate student enrolled at the Georgia Institute of } Technology. } I am in a biomedical engineering class, and we have a problem that we must } solve involving electron microscopes. I have a few questions that I hope will } get answered ASAP: } } 1. What are the advantages and disadvantages in using electrons for } microscopy } rather than light? } 2. Does the wavelength of the electrons have anythign to do with the spatial } resolution that the microscope produces in the final picture? } 3. What is temporal resolution and how is it produced in the electron } microscope? } } Thank you for your time. I greatly appreciate your efforts in helping me } understand more of this subject. } } Sincerely, } Jenny Wang } } ------------------------------------------------- } Sent through Cyberbuzz- A Server for the Students } http://cyberbuzz.gatech.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)
as you have probably noticed, 21CFR Part 11 is a pretty tough cookie, and in fact involves more than one piece of software. If you read it closely, you'll find that it involves the operating system, as well as Operating Procedures, which are outside the realm of any single software. We have found a solution to this, but I don't want to advertise this here, so please contact me off-line if you are interested.
Also, regarding the micron bars, I'd like to mention, that this is always an afterthought for Photoshop. John Russ has done a good job providing some features to put a scale bar on the image, but for a comprehensive solution, I think there are other solutions. We, for example (and other programs probably as well) make sure, that an image is calibrated from the start by either reding the calibration or magnification from the instrument or entering the magnification by hand. The scale bar is then displayed at any time in the viewport. We found, that a scale bar attached to the image or the overlay is not the best solution in many cases. For example, if you have a large image, the size of the scale bar depends on the "screen magnification": if you set that to 100%, the scale bar has a good size to see, but you may not see it because the image does not fit on the screen. If you fit the image to the screen, the scale bar may be too small to read. We tried to solve this problem by calculating the scale bar dynamically and displaying it as a part of the viewport, not the image (unless, of course, you want to attach it to the image).
Finally, I'd like to announce that we moved to new locations in Lakewood. Please make a note of our new address.
Mike Bode
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Damian Neuberger [mailto:neuberger1234-at-attbi.com] Sent: Tuesday, February 05, 2002 2:43 PM To: microscopy-at-sparc5.microscopy.com
Listers,
My requirements are that I have to put a label as well as a scale bar on each optical micrograph and burn CD to archive, all with GLP documentation (we are working towards a 21CFR Part 11 compliant system). I have calibrated our research microscope with various optical systems and photoeyepieces as well as a new macroscope and created about 100 scale bars. We then automated the whole process as much as we could using Actions and Droplets and a Word macro to create a label as well as a list of all the files burned on a CD by filling in a form.
Next we are hoping to be able to write a plug-in that will create a history text file to document each change to a photograph. I asked Adobe if they could do that and the person said that it would be feasible but apparently not enough demand, yet.
In a message dated 02/05/2002 8:42:07 AM US Mountain Standard Time, David_R_Stadden-at-armstrong.com-at-sparc5.microscopy.com writes:
{ { I've been asked if the stage micrometers I use for calibrating our optical scopes are NIST-traceable. Is there such a thing and, if not, what would satisfy requirements for ISO certification in the area of metrology?
Thanks!
Dave Stadden Research Scientist Armstrong World Industries, Inc. Lancaster, PA } }
Dave,
Yes, there is such a thing. You can get a stage micrometer that's NIST-traceable from:
If I recall correctly, you pay a minimum fee for certification of a set number of points on the scale (you can specify which points on the scale are to be certified). You can also pay a minimal fee to certify additional points above the minimum number.
SEM has historically been used for metrology of various structures. I can't seem to find much literature about the artifacts associated with this type of measurement. We do use the NIST standards to check the calibration of our equipment, but I haven't characterized how the different beam or sample parameters effect the measurements. What do you do? Has anyone figured out his or her actual accuracy and precision? We have found that we can safely give measurements within +/-5% taking into account most human and equipment errors. This is based on the precision of measurements made of NIST structures, measured the same way, over several years. On the other hand, the smallest structure we can measure on the standard is 2um (line and space. How do you determine at what magnification you will no longer guarantee the measurement? I see that my MRS-3 from Geller says it's for 10x to 50kX. How do they figure out that the max magnification it is useful? Maybe it as simple as being able to fit the structure on the screen. If that were true, you would expect that the instrument would also be calibrated to a much higher magnification. How high could I say it is accurate to? Can I safely measure a 1000A line assuming no obvious issues (i.e. drift)? Can anyone educate me more on this topic or point me to resources?
Things that could effect measurements (feel free to add to list): Drift (mechanical / beam) Charging (obvious or stretching of image from a slow scan) Magnification (adjusted for each set of lens relays) kV (surface vs. subsurface image) Working Distance Delineation method (raised vs. depression, materials contrast) Amount of delineation (3D effect) Resolution (near resolution limit of SEM?) Contrast (or lack of, bright / dark line) Edge effect (bright line) Consistency between tools (calibration, etc.) Consistency between methods (SEM, TEM, ellipsometry, AFM, etc.) Operator's eye (where to measure. Measure outside to inside, center to center, out to out, in to in?) Variance in measured layer thickness (topography, sloped profile (i.e. base larger than top)) Angle to beam Preparation methods (polish (i.e. smearing), cleave (i.e. pull of soft material), FIB (i.e. angle)) Type of algorithm if doing it automatically (i.e. %50 threshold)
_________________________________________________________________ MSN Photos is the easiest way to share and print your photos: http://photos.msn.com/support/worldwide.aspx
Yes, NIST-traceable stage micrometers do exist. At least one place they are available from is Klarmann Rulings (www.reticles.com).
We have utilized these NIST-traceable micrometers in previous systems we've implemented with much success.
Good Luck
*Kind Regards, *Dave Hall *Resolution Technology, Inc - (614) 921-0045 *Please visit our website at http://www.restechimage.com to find our Optimas training schedule and other useful information.
-----Original Message----- } From: David_R_Stadden-at-armstrong.com [mailto:David_R_Stadden-at-armstrong.com]On Behalf Of "David_R_Stadden-at-armstrong.com"-at-sparc5.microscopy.com Sent: Tuesday, February 05, 2002 10:29 AM To: Microscopy-at-sparc5.microscopy.com
I've been asked if the stage micrometers I use for calibrating our optical scopes are NIST-traceable. Is there such a thing and, if not, what would satisfy requirements for ISO certification in the area of metrology?
Thanks!
Dave Stadden Research Scientist Armstrong World Industries, Inc. Lancaster, PA
I am looking for a space in my lab for installation of 2 UPS systems (6 and 8 kVa) from Hitachi. One of the options is to install them in the same room where ultramicrotome is. I do not like this option, but space is tight. I appreciate any comments about microtome performance in the presence of UPS. Microtome is sitting on the air antivibration table.
Thank you,
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} I think your instructor's hope would be that you figured out the } answers to class problems on your own. Asking an expert in the field } and then simply regurgitating that information is a worthless } exercise. If you are going to invest the time and money required to } earn a degree, you might want to try to learn something along the } way. I am a big supporter of listservers but hate to see them used } in this way.
Good for you!!
I sent the young lady the same message via private e-mail. I wonder if the instructor knows about this sort of "research"?
} } To whom it may concern: } } } } I am an undergraduate student enrolled at the Georgia Institute of } } Technology. } } I am in a biomedical engineering class, and we have a problem that we must } } solve involving electron microscopes. I have a few questions that I hope will } } get answered ASAP: } } } } 1. What are the advantages and disadvantages in using electrons for } } microscopy } } rather than light? } } 2. Does the wavelength of the electrons have anythign to do with the spatial } } resolution that the microscope produces in the final picture? } } 3. What is temporal resolution and how is it produced in the electron } } microscope? } } } } Thank you for your time. I greatly appreciate your efforts in helping me } } understand more of this subject. } } } } Sincerely, } } Jenny Wang } } } } ------------------------------------------------- } } Sent through Cyberbuzz- A Server for the Students } } http://cyberbuzz.gatech.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)
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 would like to thank those who gave me some input to my first delimma.
I found that there are a ton of different frame grabbers out there. What I dont know is what kind of output signal the scope produces, as far as my understanding of the ISI SX-40 SEM it puts a signal out to the CRT. Some of the venders' questions have been if it is an NTSC, PAL, or RS170 type signal.I am slightly familiar with the first two, but the latter I have no idea. I do know that there are a couple of "off the shelf" products from GW Electronics and Image Slave, but these setups are priced at $4000+. So what I am looking for is if someone can point me in the direction of where I can reseach this information.
Wil
This email was sent with 100.00% recycled electrons.
P.S. This is a project that hopefully will get students involved with microcopy from our Biology and Chemistry departments in order to familiarize the students with instrumentation available in their area of study.
you could try this website of Florence University:
http://www.unifi.it/unifi/dbag/lbs1/lbsdip.htm
(sorry...it is in Italian) and mail to Dr. Stefano Bianchi at this address: stefano.bianchi-at-dbag.unifi.it
Good luck. Best Regards, Ing. Massimo Tosi ----- Original Message ----- } From: "Peter Van Osta" {pvosta-at-unionbio-eu.com} To: "Microscopy" {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, February 06, 2002 9:17 AM
Dear Lister:
I have a Epson scanner (Perfection 1200 Photo) with a transparency adaptor. Recently, we have some problems when we scan the negative films. The pre-scan image looks pretty nice after some adjustment, but the final scan image is too dark almost just a piece of black stuff. Any help will be appreciated.
Nikon Coolpix 995 has 2.3Megapixels non-interpolated
Tina Carvalho {tina-at-pbrc.hawaii.edu} wrote:
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} I operate a JEOL 848 SEM but am limited to less than 30,000x due to at least } two frequencies of external noise. The service engineers from JEOL have } determined the noise is not due to the SEM. I will be conducting as much of a } room survey as possible later in February but I am hoping to obtain some } insight into common testing formats, noise sources, and solutions. From what } I have been able to learn so far, this noise reduction is something of an art } rather than science. Thank you in advance from this microscopist with much to } learn.
I agree that the other responses have very good points.
In addition, if the problem is actually caused by magnetic fields interacting with the beam, it should get worse at lower kV and at longer working distances. Do you know the frequencies of the interference? If so, that can be a very good clue as to its source.
Also, a digital gauss meter that I have found very useful for locating sources of magnetic fields can be purchased for under $100 US [see the Extech Model #480823 at www.MetersandInstruments.com, (800) 773-0370, - I have no financial interest in this company]. This meter makes frequency independent RMS readings between 30 and 300 Hz. Since line frequency and harmonics are the most common fields, it works very well.
With such a digital meter, or even a coil of wire connected to an oscilloscope to act as an uncalibrated magnetic field meter, you can first check for field strength near the chamber. If a significant magnetic field (i.e., } 1 mG rms) is observed, it can often be tracked back to the source by moving the meter around. If the field simply "fills the room", it may be from a power line with an unbalanced load. For example, if the current runs on the "hot" wire, but does not return on the neutral or ground wire, then a large magnetic field will be generated, while normally there are equal and opposite currents which cancel at any significant distance. In this case, the wires themselves can be outside the room and the actual wiring problem may be anywhere in the building!
As a final comment, a good check on environmental problems is simply to come in after hours when other equipment is more likely to be turned off (or to go around and turn off everything that you can, but that is typically limited to your immediate lab). For example, I have watched a gauss meter go from showing a low field to a significant field simultaneously as an image at high mag degraded with line frequency interference. The source of the field was discovered to be power lines outside the SEM room that supplied power to a laser several labs away. When the laser operator started working in the morning and increased the laser power, the meter and SEM display clearly showed the interference.
Good luck.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
**************************************************************************** * 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* ****************************************************************************
} -----Original Message----- } From: Brian Wajdyk [mailto:electronmicroscopist-at-hotmail.com] } Sent: Wednesday, February 06, 2002 10:47 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: measurement and calibration onthe SEM } } } -------------------------------------------------------------- } ---------- } 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 } -------------------------------------------------------------- } ---------. } } } Fellow Microscopists, } } SEM has historically been used for metrology of various } structures. I can't } seem to find much literature about the artifacts associated } with this type } of measurement. We do use the NIST standards to check the } calibration of } our equipment, but I haven't characterized how the different } beam or sample } parameters effect the measurements. What do you do? Has } anyone figured out } his or her actual accuracy and precision? We have found that } we can safely } give measurements within +/-5% taking into account most human } and equipment } errors. This is based on the precision of measurements made of NIST } structures, measured the same way, over several years. On } the other hand, } the smallest structure we can measure on the standard is 2um } (line and } space. How do you determine at what magnification you will no longer
Cheap grating replicas with 2600 lines per mm give 0.46 um per line, and this is not too bad for calibrating 50,000 magnification. I am happy I do not need certified standards.
} guarantee the measurement? I see that my MRS-3 from Geller } says it's for } 10x to 50kX. How do they figure out that the max magnification it is } useful? Maybe it as simple as being able to fit the structure on the
Maximum useful magnification is very specimen dependant, especially for low voltage and low vacuum modes. Of course, for digital images it is possible to check brightness profiles and if they have slopes on edges of features, then measure "size" on half height of the slope. But I am not aware about publications which dependably justify this kind of measurements (manipulations with brightness and contrast and specimen tilt could change slopes significantly).
} screen. If that were true, you would expect that the } instrument would also } be calibrated to a much higher magnification. How high could } I say it is } accurate to? Can I safely measure a 1000A line assuming no
It depends on resolution for your microscope/specimens and on calibration standard you are using. And I think periodic lines with spacing 2 um not really good standard to measure feature with the size of 0.1 um.
} obvious issues } (i.e. drift)? Can anyone educate me more on this topic or
If you have visible drift during single exposure, then something wrong with microscope or specimen preparation technique.
} point me to } resources? } } } Things that could effect measurements (feel free to add to list): } Drift (mechanical / beam)-
exposure time should be small for significant drift.
} Charging (obvious or stretching of image from a slow scan) } Magnification (adjusted for each set of lens relays)
Could be eliminated with proper calibration.
} kV (surface vs. subsurface image) } Working Distance
Could be eliminated with proper calibration.
} Delineation method (raised vs. depression, materials contrast) } Amount of delineation (3D effect) } Resolution (near resolution limit of SEM?) } Contrast (or lack of, bright / dark line) } Edge effect (bright line) } Consistency between tools (calibration, etc.) } Consistency between methods (SEM, TEM, ellipsometry, AFM, etc.) } Operator's eye (where to measure. Measure outside to inside, } center to } center, out to out, in to in?) } Variance in measured layer thickness (topography, sloped } profile (i.e. base } larger than top)) } Angle to beam } Preparation methods (polish (i.e. smearing), cleave (i.e. } pull of soft } material), FIB (i.e. angle)) } Type of algorithm if doing it automatically (i.e. %50 threshold)
Some of the things you have mentioned relate to specimen/experiment, to stereology, but not to microscope. For example, if I need to measure size of depression without sharp edges, I have to find (or at least to declare)right procedure for it's measurements. May be I have to perform stereo measurements and define an edge as a place, where a depth of depression become equal to 0.1 um (or 10% of total depth, or whatever else, depending on a study).
And thank you for your extensive list - it is very helpful for observation of the problem. And about additions to your list - I think everybody can say something. For example recently I tried to measure in ESEM thickness of a layer which, as it turned out, was a viscous liquid...
Regards,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
On Wed, 6 Feb 2002 tartenon-at-netscape.net-at-sparc5.microscopy.com wrote:
} Nikon Coolpix 995 has 2.3Megapixels non-interpolated
You made me check into this! Nikon advertises 3.2 megapixels for the Coolpix 995, and Olympus advertises 4 megapixels for the C-4040, but BOTH really deliver 2048 x 1536 pixels. Interesting marketing.
They are basically the same technology, with the only difference being the menus and availability of adapters, I guess. And now all the adapters are available all kinds of places, so I guess it's a matter of taste. There aer other manini (a manini is a small fish) differences in number of threads, recording media, stuff like that.
Mahalo, Tina
**************************************************************************** * 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* ****************************************************************************
We have a Hitachi H800 TEM (200 Kev,LaB6) and a few days ago we started having problems with the HV. Static discharges in the gun area (even at 75 KeV) were traced back to the failed insulator. Hitachi is trying to get us new one, but they tell me it could take several months since this is normally not a stock item. If any body out there has one that they are willing to part with (for money) or if any body has other suggestions I would really appreciate your input.
RS-170 refers to composite b/w video. NTSC refers to composite color video added to the RS-170 format. Composite means that the video signal and sync pulses (H & V & blanking) are conveyed via a single wire.
If I recall the SX-40 correctly (let me know if I am wrong on this), it has a TV rate display for focusing and stig and eye-ball viewing, but image recording is/was via the slow scan high rez short persistence 2K line film recorder CRT. This poses a significant problem for image capture.
RS-170 video is interlaced between even/odd lines (fields). One frame (two fields) is not presented at the same time. The rapid scan (60/s per field) and the persistence of the viewing CRT, fool the eye into being able to see integrated sets of lines. A frame grabber can't be fooled. The purpose of interlacing is to reduce flicker which would occur if the whole frame were sent at one time.
The RS-170 format is essentially a frame of 640x480 pixels. But the problem is that to capture the whole frame, one needs to store the even and odd fields and then grab the frame. This feature is typically called an image buffer or frame buffer. Its output is RS-170 but consisting of a full frame, either realtime or the result of slow scan.
I don't think you have this in the SX-40. Your only option, as I see it, is to get a passive capture system which is attached to the recording CRT. The passive capture systems connect to the recording CRT's H & V sync pulses and the blanking pulse. GW makes passive capture systems, Soft Imaging does, as do others. But as you have found, these are not cheap systems. In some cases, the hardware is not particularly complex. But the software is.
The passive system follows the scan pulses from the SEM's scan generator and uses an A/D converter to sequentially digitize the output of the SE detector. A challenge with this is to obtain fidelity between what is seen versus what is captured. i.e., same contrast and brightness on the viewing screen as on the captured image. Doable, once set up properly.
Perhaps you could elicit the assistance of the Electrical Engineering Department? Some clever grad student might love to undertake a project to digitize your SEM.
gary g.
At 10:27 AM 2/6/2002, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Wil,
the answer is fairly simple:
A "video signal" is commonly referred to if the signal follows the standards set for commercial TV. This allows you to take a camera and connect it directly to a TV or VCR. As you probably know, there are different standards in the US and Europe. NTSC is the signal used for color data in the US and Japan, PAL is the signal used for color data in Europe and other places, RS170 is for b/w signals. Some frame grabbers can understand all of these signals, others only one or two.
The problem with a video signal is, that the resolution is very low (480x640x8bits for NTSC).
If you use the SEM in "slow scan mode", most frame grabbers will not be able to detect the correct signal because it does not conform to the NTSC or PAL standards anymore. Then you need to get special electronics, which is not made in large numbers and is therefore more expensive.
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: "curari-at-asu.edu"-at-sparc5.microscopy.com [mailto:"curari-at-asu.edu"-at-sparc5.microscopy.com] Sent: Wednesday, February 06, 2002 11:27 AM To: MSA
Dear Lister
I would like to thank those who gave me some input to my first delimma.
I found that there are a ton of different frame grabbers out there. What I dont know is what kind of output signal the scope produces, as far as my understanding of the ISI SX-40 SEM it puts a signal out to the CRT. Some of the venders' questions have been if it is an NTSC, PAL, or RS170 type signal.I am slightly familiar with the first two, but the latter I have no idea. I do know that there are a couple of "off the shelf" products from GW Electronics and Image Slave, but these setups are priced at $4000+. So what I am looking for is if someone can point me in the direction of where I can reseach this information.
Wil
This email was sent with 100.00% recycled electrons.
P.S. This is a project that hopefully will get students involved with microcopy from our Biology and Chemistry departments in order to familiarize the students with instrumentation available in their area of study.
From root Wed Feb 6 21:55:53 2002 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id VAA12231 for dist-Microscopy; Wed, 6 Feb 2002 21:01:57 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id VAA12227 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 6 Feb 2002 21:01:26 -0600 (CST) Received: from server.imre.org.sg ([137.132.173.15]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id UAA12178 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 6 Feb 2002 20:49:18 -0600 (CST) Message-ID: {FEBD650DBBBFD311AF1D009027CC7DD701366A1A-at-exchange.imre.org.sg}
Dear Jenny,
We're glad that you are interested in microscopy, and that you did some research to find this list: below are a few pointers to get you started. There are some excellent texts on the subject, some of which I cite below from my own lecture course, but check out your own library too. Also, there are some very well known electron microscopists at Georgia Tech whom you could consult with - be brave and go say hi! You will find the majority of scientists (microscopists included) are always glad of an excuse to wax eloquent on their favorite subject, esp over coffee. I guess the best advice is always to talk to the experts, unless forbidden by your instructor (which I would doubt - our instructors always encouraged our intiative in these matters!)
Best wishes for your studies,
Mark
%%%%%%%%%%%%%%%%%% Mark Yeadon Senior Research Fellow Institute of Materials Research and Engineering 3 Research Link Singapore 117602
Assistant Professor Department of Materials Science National University of Singapore Singapore 119260 http://www.matsci.nus.edu.sg/STAFF/Mark.html
} -----Original Message----- } From: "gtg990a-at-prism.gatech.edu"-at-sparc5.microscopy.com } [SMTP:"gtg990a-at-prism.gatech.edu"-at-sparc5.microscopy.com] } Sent: Wednesday, February 06, 2002 11:14 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Questions on the Electron Microscope } } ------------------------------------------------------------------------ } 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 whom it may concern: } } I am an undergraduate student enrolled at the Georgia Institute of } Technology. } I am in a biomedical engineering class, and we have a problem that we must } } solve involving electron microscopes. I have a few questions that I hope } will } get answered ASAP: } } 1. What are the advantages and disadvantages in using electrons for } microscopy } rather than light? [Mark YEADON] couple of advantages: electrons have a shorter wavelength (much shorter); since resolution in this case is dependent on wavelength (Abbe's expression - resolution ~ 0.61 x wavelength / sin alpha [see texts for fuller explanation]) - in principle we can resolve distances of a fraction of an atom with 100kV electrons). Also, electrons are IONIZING radiation, and we can detect signals arising from ionization processes when they interact with our sample (such as characteristic x-rays, charcteristic of the atoms the electron has interacted with in your sample)
couple of disadvantages: electrons require a vacuum if you want them to travel enough distance to be of use in an electron microscope. this is expensive and requires substantial maintenance. electron guns and columns are much bigger and more complex than a regular light microscope - equates to dollars!
However, because the value of alpha (in above equation) is so much smaller for electron microscopes you also get an amazingly high depth of field (see texts below for a derivation - esp the first three). ie, you can see 3D objects such as bugs in clear focus over the entire sample in a scanning electron microscope with marvelous resolution, whereas in the light microscope only one small part of the bug will be in focus at high magnification...
} 2. Does the wavelength of the electrons have anythin to do with the } spatial } resolution that the microscope produces in the final picture? [Mark YEADON] Absolutely - see above.
} 3. What is temporal resolution and how is it produced in the electron } microscope? } [Mark YEADON] Spatial resolution relates to dimensions of distance, for a 200kV TEM with thermionic emission gun you would expect about 2Angstroms spatial resolution. Temporal resolution would relate to time, and I'm guessing you're thinking of the time taken to capture images - this would depend upon the imaging system you are using. How quickly can you record 'frames', one after the other. We get 1/30s temporal resolution for in-situ experiments quite ok. In principle you can go much better than this with a very fast CCD camera and a high intensity electron beam. (The more electrons you have per unit time, the more electrons you can get per frame, and the better the signal to noise ratio - the limit is usually the recording equipment (signal to nosie ratio of the CCD chip) and not the TEM.
} Thank you for your time. I greatly appreciate your efforts in helping me } understand more of this subject. [Mark YEADON] You're most welcome. We love our subject and are delighted to help you in your understanding. See if you have any of the following in your library, although there are many other good ones also:
Transmission Electron Microscopy, DB Williams and CB Carter Electron Microscopy and Analysis, Goodhew and Humphreys Light and Electron Microscopy, Slayter and Slayter Handbook Of Microscopy, ed. by S. Amelinckx et al., Wiley -VCH
} Sincerely, } Jenny Wang } } ------------------------------------------------- } Sent through Cyberbuzz- A Server for the Students } http://cyberbuzz.gatech.edu/
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pjfenneran-at-msn.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, February 6, 2002 at 10:05:13 ---------------------------------------------------------------------------
Email: pjfenneran-at-msn.com Name: Patrick Fenneran
Organization: Florida Institute of Technology
Education: Graduate College
Location: Melbourne, Florida
Question: I am using an Zeiss 900m TEM looking at E.coli and have the following questions:
1. The formvar/carbon grids keep on getting blown out, like there is too much power or the beam is too concentrated.
2. When I am taking pictures, the bacteria do not have resolution, which adjustments should I make?
3. Do you know of any paperwork that can be obtained that explains the operation the components of the machine, the only book I have is the one that came with the machine and it is partly in German.
It depends exactly what you mean by 'failed' but we have had success cleaning ceramic insulators that have been subjected to tracking of the HT. Even quite deep arcing tracks have been cleaned out and when returned to service have worked OK. If they are on the vacuum side they are just left alone, if they are on a greased joining surface we ensure that the grease gets well down into the cleaned track.
I don't know the type of insulator Hitachi use on your machine but you may be able clean tracks out by polishing with Al2O3 paste (5um Al2O3 in alcohol). If this does not work try shot blasting the ceramic with clean Al2O3 beads. After polishing or blasting blow off with clean N2 (from a regulated cylinder), clean in several washes of alcohol in an ultrasonic bath until there is no trace of alumina in the alcohol and bake out in a clean vacuum oven overnight. If you cannot get a vacuum oven then heating in air and a very long pumpout in a vacuum rig may work. It will take a couple of days but it may get you up and running and may even save you buying a replacement insulator.
Disclaimer: I take no responsibility for people not applying common sense to any of the procedures descibed. If you do not have good technical experience and skills you are likely to cause further damage.
Good luck, Ron
On Wed, 6 Feb 2002 14:55:44 -0700 "Marti, Jordi" {jordi.marti-at-honeywell.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 } -----------------------------------------------------------------------. } } } Hello! } } We have a Hitachi H800 TEM (200 Kev,LaB6) and a few days ago we started having problems with the HV. Static discharges } in the gun area (even at 75 KeV) were traced back to the failed insulator. Hitachi is trying to get us new one, but they } tell me it could take several months since this is normally not a stock item. If any body out there has one that they } are willing to part with (for money) or if any body has other suggestions I would really appreciate your input. } } Thanks } } Jordi Marti }
---------------------- 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
We have blown the window on our old Noran EDS detector. I would like to find out an approximate price on a new system within about 10k. We would like to replace the old one. Before we get the formal quotes, I though a few of you that have recently purchased systems could give me a ballpark price. Thanks,
Mark Windland Honeywell Minneapolis, Minnesota 763-954-2845
I read Ms Wang's message, thought the same thoughts you articulated here, and deleted the message. I think that we as a group should agree not to do homework for students. Thanks for sharing your thoughts with us and raising the issue.
Don
On Wed, 6 Feb 2002, Tom Phillips 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 think your instructor's hope would be that you figured out the } answers to class problems on your own. Asking an expert in the field } and then simply regurgitating that information is a worthless } exercise. If you are going to invest the time and money required to } earn a degree, you might want to try to learn something along the } way. I am a big supporter of listservers but hate to see them used } in this way. } } } } } } } } To whom it may concern: } } } } I am an undergraduate student enrolled at the Georgia Institute of } } Technology. } } I am in a biomedical engineering class, and we have a problem that we must } } solve involving electron microscopes. I have a few questions that I hope will } } get answered ASAP: } } } } 1. What are the advantages and disadvantages in using electrons for } } microscopy } } rather than light? } } 2. Does the wavelength of the electrons have anythign to do with the spatial } } resolution that the microscope produces in the final picture? } } 3. What is temporal resolution and how is it produced in the electron } } microscope? } } } } Thank you for your time. I greatly appreciate your efforts in helping me } } understand more of this subject. } } } } Sincerely, } } Jenny Wang } } } } ------------------------------------------------- } } Sent through Cyberbuzz- A Server for the Students } } http://cyberbuzz.gatech.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) } }
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
We had the same problem with the ceramic insulator (sounds like a law of nature...). A big help in our case was a dentist. He has much experience in cleaning ceramic and the perfect tools for doing this job. So my advice is to bring the part to be cleaned to a dentist in your neighborhood.
email: mklein-at-visitec-em.de WWW: http://www.visitec-em.de ++++ Home of the world`s largest SEM ++++
-----Ursprüngliche Nachricht----- Von: rdoole-at-materials.ox.ac.uk [mailto:rdoole-at-materials.ox.ac.uk]Im Auftrag von Ron Doole Gesendet: Donnerstag, 7. Februar 2002 09:49 An: 'Microscopy' Betreff: Re: Ceramic Insulator for Hitachi H800 TEM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Jordi,
Sorry to hear about the insulator.
It depends exactly what you mean by 'failed' but we have had success cleaning ceramic insulators that have been subjected to tracking of the HT. Even quite deep arcing tracks have been cleaned out and when returned to service have worked OK. If they are on the vacuum side they are just left alone, if they are on a greased joining surface we ensure that the grease gets well down into the cleaned track.
I don't know the type of insulator Hitachi use on your machine but you may be able clean tracks out by polishing with Al2O3 paste (5um Al2O3 in alcohol). If this does not work try shot blasting the ceramic with clean Al2O3 beads. After polishing or blasting blow off with clean N2 (from a regulated cylinder), clean in several washes of alcohol in an ultrasonic bath until there is no trace of alumina in the alcohol and bake out in a clean vacuum oven overnight. If you cannot get a vacuum oven then heating in air and a very long pumpout in a vacuum rig may work. It will take a couple of days but it may get you up and running and may even save you buying a replacement insulator.
Disclaimer: I take no responsibility for people not applying common sense to any of the procedures descibed. If you do not have good technical experience and skills you are likely to cause further damage.
Good luck, Ron
On Wed, 6 Feb 2002 14:55:44 -0700 "Marti, Jordi" {jordi.marti-at-honeywell.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 } -----------------------------------------------------------------------. } } } Hello! } } We have a Hitachi H800 TEM (200 Kev,LaB6) and a few days ago we started having problems with the HV. Static discharges } in the gun area (even at 75 KeV) were traced back to the failed insulator. Hitachi is trying to get us new one, but they } tell me it could take several months since this is normally not a stock item. If any body out there has one that they } are willing to part with (for money) or if any body has other suggestions I would really appreciate your input. } } Thanks } } Jordi Marti }
---------------------- 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
I have just been given the approval to purchase a new carbon coater. We are looking for a model that doesn't use a water source. We have an existing Denton 502A that we are going to change to a backup system. It has been a workhorse for us for the past 14 years.(Up and running six hours a day.)
At your lab, what has been a proven carbon coater that will last?
I need to submit three vendors to my boss before he will approve a purchase of a new carbon coater. As you can see I have been out of the loop on carbon coaters for fourteen years so any information will help me tremendously.
Thank you for your help.
Luis Bustillos AMA Analytical Services, Inc. lbustillos-at-amalab.com
} } Dear Lister: } } I have a Epson scanner (Perfection 1200 Photo) with a transparency } adaptor. Recently, we have some problems when we scan the negative } films. The pre-scan image looks pretty nice after some adjustment, } but the final scan image is too dark almost just a piece of black } stuff. } Any help will be appreciated. } } Thanks } } Jinguo Wang ******** I have an Epson Expression 1600 and had the same problem. My solution was to select "TPU for Pos film" which does not do the automatic inversion. I then use the Invert function in Photoshop to reverse the contrast from negative to positive. It works beautifully. Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
About a year ago there was a valuable discussion of negative scanners. One concern raised about the Polaroid Sprintscan 45 Ultra was the absence of a film holder for 3 1/4 x 4 1/4 film. A message was sent that said that John Warren at Polaroid sent several of you free negative holders.
Last year I purchased this scanner and included a 3 1/4 x 4 1/4 negative holder on the P.O., and have been waiting for the holder. Polaroid told my sales rep that such a holder never existed and that John Warren no longer worked a polaroid.
I have (and love) the Polaroid scanner, but am frustrated about not having the appropriate film holder. Have any of you received this item, does it have a part number, and how did you come to have it?
Any help would be appreciated.
Thanks,
Don
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
The Microbeam Analysis Society's home page has a new url, http://www.microbeamanalysis.org. We apologize for this inconvience and I want to thank John Mansfield for his prompt attention in establishing our new domain name. If you are a subscriber to the MAS listeserver, John autimatically changed your subscription to the new address at microprobe-at-microbeamanalysis.org.
Also, the MAS membership email service (masmembership-at-excite.com ) was interrupted for a few weeks in January but is fully functional again.
Lou Ross MAS Membership Services masmembership-at-excite.com 1-800-4MASMEM (1-800-462-7636) www.microbeamanalysis.org
Sounds to me like both points 1. and 2. could be explained if the condenser aperture is out of the column. Its the uppermost adjustable aperture sticking out of the side of most TEM's. If thats out, you get huge beam current down the column, big loss of resolution, and its easy to blow out the sections or support films.
Or even if the condenser aperture is in, if the next adjustable aperture down the column, the objective aperture, is out, that also could result in blown out films, and loss of contrast in the image.
As for the German, best to find someone there who can give you a few pointers - in English,
Good luck,
Gib
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
} Email: pjfenneran-at-msn.com } Name: Patrick Fenneran } } Organization: Florida Institute of Technology } } Education: Graduate College } } Location: Melbourne, Florida } } Question: I am using an Zeiss 900m TEM looking at E.coli and have the } following questions: } } 1. The formvar/carbon grids keep on getting blown out, like there is } too much power or the beam is too concentrated. } } 2. When I am taking pictures, the bacteria do not have resolution, } which adjustments should I make? } } 3. Do you know of any paperwork that can be obtained that explains } the operation the components of the machine, the only book I have is } the one that came with the machine and it is partly in German. }
We have progressed during the past few years to where the majority of our optical and SEM images are digital rather than Polaroid prints. In addition, we also scan TEM negatives and store them digitally. I am interested in some recommendations for software that can store the images in a database so they can be easily retrieved by keywords and also software for image analysis (e.g., particle size, image analysis, etc.) What options are available that people have experience with. I'd be interested in hearing both pro and con.
Thanks,
Bob Comstock Westinghouse Electric Co. Pittsburgh, PA 15235
Were the bacteria fixed and stained? Did you carbon coat your formvar grids before use? What mesh size grids are you using? I have no information on that particular microscope, but if you have never used any TEM, you normally use a spread weak beam rather than a concentrated one for imaging (spread the beam with the condensor lense). Electron microscopes work best in the dark.
We have some procedures on our website you may wish to browse: http://biology.berkeley.edu/EML
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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
On Thu, 7 Feb 2002 pjfenneran-at-msn.com wrote: } 1. The formvar/carbon grids keep on getting blown out, like there is } too much power or the beam is too concentrated. } } 2. When I am taking pictures, the bacteria do not have resolution, } which adjustments should I make? } } 3. Do you know of any paperwork that can be obtained that explains } the operation the components of the machine, the only book I have is } the one that came with the machine and it is partly in German. } } } } --------------------------------------------------------------------------- }
Sorry You're Right I miss type the numbers it really is 3.2 Megapixels where 2048 X 1536 = 3,145,728 (3.2 Megapixels) I'm using Nikon 995 and I can Photograph Bright, Fluorescence Images My Understanding is that you can increase that resolution using Interpolation, but the real resolution of the image will be 3.2 Megapixels with the Nikon 995. I do not belive Olympus has 4 Megapixels non-interpolated (nor any available digital camera)
Regards
Tina Carvalho {tina-at-pbrc.hawaii.edu} wrote:
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} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } Tom: } } I read Ms Wang's message, thought the same thoughts you articulated here, } and deleted the message. I think that we as a group should agree not to } do homework for students. Thanks for sharing your thoughts with us and } raising the issue. } } Don } } On Wed, 6 Feb 2002, Tom Phillips wrote: } } } } I think your instructor's hope would be that you figured out the } } answers to class problems on your own. Asking an expert in the field } } and then simply regurgitating that information is a worthless } } exercise. If you are going to invest the time and money required to } } earn a degree, you might want to try to learn something along the } } way. I am a big supporter of listservers but hate to see them used } } in this way. } } } } } } } } To whom it may concern: } } } } } } I am an undergraduate student enrolled at the Georgia Institute of } } } Technology. } } } I am in a biomedical engineering class, and we have a problem that we must } } } solve involving electron microscopes. I have a few questions that I hope will } } } get answered ASAP: } } } } } } 1. What are the advantages and disadvantages in using electrons for } } } microscopy } } } rather than light? } } } 2. Does the wavelength of the electrons have anythign to do with the spatial } } } resolution that the microscope produces in the final picture? } } } 3. What is temporal resolution and how is it produced in the electron } } } microscope? } } } } } } Thank you for your time. I greatly appreciate your efforts in helping me } } } understand more of this subject. } } } } } } Sincerely, } } } Jenny Wang } } } } } } ------------------------------------------------- } } } Sent through Cyberbuzz- A Server for the Students } } } http://cyberbuzz.gatech.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) } } } } } } ______________________________________________________________________ } Donald L. Lovett e-mail: lovett-at-tcnj.edu } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } P.O. Box 7718 fax: (609) 637-5118 } The College of New Jersey } Ewing, NJ 08628-0718
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 **********************************************
From root Thu Feb 7 13:17:04 2002 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id NAA15504 for dist-Microscopy; Thu, 7 Feb 2002 13:05:04 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id NAA15501 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 7 Feb 2002 13:04:34 -0600 (CST) Received: from tumor.soft-imaging.com ([67.104.115.34]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id NAA15494 for {Microscopy-at-sparc5.Microscopy.Com} ; Thu, 7 Feb 2002 13:04:18 -0600 (CST) Received: from lakewood.soft-imaging.com (lakewood.soft-imaging.com [192.168.5.225]) by tumor.soft-imaging.com (8.10.2/8.10.2/SuSE Linux 8.10.0-0.3) with ESMTP id g17J4jr07268 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 7 Feb 2002 12:04:46 -0700 Received: by hq-dc2.soft-imaging.com with Internet Mail Service (5.5.2653.19) id {DRCSJ20P} ; Thu, 7 Feb 2002 12:03:40 -0700 Message-ID: {6127CE87B9BDD511B59D0001028A497D03E2A3-at-hq-dc2.soft-imaging.com}
Hello Listers,
While I agree with what has been said so far in that nobody should do somebody else's work, especially not homework for students, I am wondering if that is not being interpreted a bit too narrowly here. After all, this listserver **IS** a great resource and the students show some initiative in finding and posting to it. Wouldn't it be better to help the students find the information they are after rather than denying help? Something like: "These are very interesting questions, and there are many good books about this issue. Try reading any one of these books (...) or talk to anyone who is doing electron microscopy."
Then again, these are my thoughts, and if you disagree, please send me an email.
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Donald Lovett [mailto:lovett-at-tcnj.edu] Sent: Thursday, February 07, 2002 6:32 AM To: Tom Phillips Cc: "gtg990a-at-prism.gatech.edu"-at-sparc5.microscopy.com; Microscopy-at-sparc5.microscopy.com
Tom:
I read Ms Wang's message, thought the same thoughts you articulated here, and deleted the message. I think that we as a group should agree not to do homework for students. Thanks for sharing your thoughts with us and raising the issue.
Don
On Wed, 6 Feb 2002, Tom Phillips 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 think your instructor's hope would be that you figured out the } answers to class problems on your own. Asking an expert in the field } and then simply regurgitating that information is a worthless } exercise. If you are going to invest the time and money required to } earn a degree, you might want to try to learn something along the } way. I am a big supporter of listservers but hate to see them used } in this way. } } } } } } } } To whom it may concern: } } } } I am an undergraduate student enrolled at the Georgia Institute of } } Technology. } } I am in a biomedical engineering class, and we have a problem that we must } } solve involving electron microscopes. I have a few questions that I hope will } } get answered ASAP: } } } } 1. What are the advantages and disadvantages in using electrons for } } microscopy } } rather than light? } } 2. Does the wavelength of the electrons have anythign to do with the spatial } } resolution that the microscope produces in the final picture? } } 3. What is temporal resolution and how is it produced in the electron } } microscope? } } } } Thank you for your time. I greatly appreciate your efforts in helping me } } understand more of this subject. } } } } Sincerely, } } Jenny Wang } } } } ------------------------------------------------- } } Sent through Cyberbuzz- A Server for the Students } } http://cyberbuzz.gatech.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) } }
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
Greetings, I would like to find a distributor for Cargille Laser Liquids (series 5610). I would like to do some experiments involving immersion oils of varied RI in combination with mountants of differing RI in a manner similar to the method employed in:
Scalettar, Swedlow, Sedat & Agard. 1996. Dispersion, abberation and deconvolution in multi-wavelength fluorescence images. Journal of Microscopy. 182:1, 50-60.
Any advice regarding sources of reagents and techniques to adjust the RI of immersion media appropriate for LSCM/MP is welcome. Thanks in advance. -Karl G.
_______________________________________________ Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illiniois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Room B650J Tel: (217) 244-6292 Fax: (217) 244-6219 www.itg.uiuc.edu
From root Thu Feb 7 14:31:53 2002 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id OAA15738 for dist-Microscopy; Thu, 7 Feb 2002 14:26:25 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id OAA15734 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 7 Feb 2002 14:25:54 -0600 (CST) Received: from mante65.nam.dow.com (mail1.dow.com [216.99.65.22]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id OAA15727 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 7 Feb 2002 14:25:43 -0600 (CST) Received: by mante65.nam.dow.com with Internet Mail Service (5.5.2653.19) id {D8514XLZ} ; Thu, 7 Feb 2002 15:21:41 -0500 Message-ID: {F98016EDC45DD411935900D0B781F18C04D19B2F-at-MANTE69}
Bob: You're likely to get a ton of vendor responses on this one, but I'll throw in a couple of general thoughts.
Disclaimer: the commercial products mentioned here are EXAMPLES. This use does not constitute any endorsement by myself or Dow Chemical or imply anything about the quality or applicability of the product. It also does not imply anything about the competitive products that are NOT mentioned.
First off, for image analysis, you need to keep ImageJ (http://rsb.info.nih.gov/ij/) in mind. It is public domain, is fully customizable (using Java language) has a wide range of users/developers around the world and works nicely. The user community is doing a lot of cool stuff, much of which gets back onto the ImageJ download page and is available for everyone else. This is the next generation of NIH Image, but it is now a Java app and runs well on MacOS, Windows, UNIX, Linux and any other OS that supports Java. You are likely to want something commercial for your "turnkey" stuff, like Image Pro+ (http://www.mediacy.com/), but ImageJ is a great tool to keep handy.
As for database/archive, I encourage you to do some math: How many images do you collect per year? How much additional information do you need to keep with those images in order for them to be useful? How many times do you need those images back after the project is complete? How long do you want to keep the images in on-line storage? How much on-line storage are you able to afford? Who will be administering the database and how much will that administration cost?
We deal with the issue by using our LIMS project number, then burning a CD(s) with all the images related to that project so that we can pull out the CD to retrieve the images. We leave the images on the network file server until the project is complete, then burn the CD and delete the on-line files.
If we have "definitive" or reference images, we keep those in on-line storage. These are the ones that really need to be part of a database.
One tool that has been useful is Thumbs+ (http://www.cerious.com/). We can make an image directory of the project CDs, then browse that to see if we can find the image we want. Thumbs+ is really aimed at a "smaller" operation of just a few people. One company that I know of with a product for maintaining a large-scale database is Advanced Database Systems (http://www.adsdb.com/), but there are others out there as well. Cost is proportional to scale!
Bill William A. Heeschen, Ph.D. The Dow Chemical Company Microscopy, Digital Imaging 1897 Bldg, E-84 / 2040 Bldg, 1330 waheeschen-at-dow.com
-----Original Message----- } From: Comstock, Robert J. [mailto:comstorj-at-westinghouse.com] Sent: Thursday, February 07, 2002 11:43 AM To: MicroscopyListserver (E-mail)
We have progressed during the past few years to where the majority of our optical and SEM images are digital rather than Polaroid prints. In addition, we also scan TEM negatives and store them digitally. I am interested in some recommendations for software that can store the images in a database so they can be easily retrieved by keywords and also software for image analysis (e.g., particle size, image analysis, etc.) What options are available that people have experience with. I'd be interested in hearing both pro and con.
Thanks,
Bob Comstock Westinghouse Electric Co. Pittsburgh, PA 15235
From root Thu Feb 7 16:17:32 2002 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id QAA15976 for dist-Microscopy; Thu, 7 Feb 2002 16:02:05 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id QAA15973 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 7 Feb 2002 16:01:35 -0600 (CST) Received: from postal1.lbl.gov (postal1.lbl.gov [128.3.7.82]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id PAA15936 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 7 Feb 2002 15:47:02 -0600 (CST) Received: from SpamWall.lbl.gov (localhost [127.0.0.1]) by postal1.lbl.gov (8.11.2/8.11.2) with ESMTP id g17Lh0E25008 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 7 Feb 2002 13:43:00 -0800 (PST) Received: from lbl.gov (maokeefe.wins.lbl.gov [131.243.3.225]) by SpamWall.lbl.gov (8.11.2/8.11.2) with ESMTP id g17LgxJ25002; Thu, 7 Feb 2002 13:42:59 -0800 (PST) Message-ID: {3C62F5F8.2463A63A-at-lbl.gov}
Don:
I'm afraid I disagree with you and Tom. Part of a scientist's training is to learn how to identify and use resources. And we (collectively) are a resource that I believe should be made available to all who can benefit. Of course, that leaves open
the question of whether the student benefits more by working things out in isolation,
or by seeking guidance from experts and either really understanding the answers or (hopefully not) merely regurgitating them by rote. One hopes that the intelligent student will reject the latter course.
Mike
Donald Lovett 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 } -----------------------------------------------------------------------. } } Tom: } } I read Ms Wang's message, thought the same thoughts you articulated here, } and deleted the message. I think that we as a group should agree not to } do homework for students. Thanks for sharing your thoughts with us and } raising the issue. } } Don } } On Wed, 6 Feb 2002, Tom Phillips 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 think your instructor's hope would be that you figured out the } } answers to class problems on your own. Asking an expert in the field } } and then simply regurgitating that information is a worthless } } exercise. If you are going to invest the time and money required to } } earn a degree, you might want to try to learn something along the } } way. I am a big supporter of listservers but hate to see them used } } in this way. } } } } } } } } } } } } } To whom it may concern: } } } } } } I am an undergraduate student enrolled at the Georgia Institute of } } } Technology. } } } I am in a biomedical engineering class, and we have a problem that we must } } } solve involving electron microscopes. I have a few questions that I hope will } } } get answered ASAP: } } } } } } 1. What are the advantages and disadvantages in using electrons for } } } microscopy } } } rather than light? } } } 2. Does the wavelength of the electrons have anythign to do with the spatial } } } resolution that the microscope produces in the final picture? } } } 3. What is temporal resolution and how is it produced in the electron } } } microscope? } } } } } } Thank you for your time. I greatly appreciate your efforts in helping me } } } understand more of this subject. } } } } } } Sincerely, } } } Jenny Wang } } } } } } ------------------------------------------------- } } } Sent through Cyberbuzz- A Server for the Students } } } http://cyberbuzz.gatech.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) } } } } } } ______________________________________________________________________ } Donald L. Lovett e-mail: lovett-at-tcnj.edu } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } P.O. Box 7718 fax: (609) 637-5118 } The College of New Jersey } Ewing, NJ 08628-0718
In a message dated 2/7/02 12:22:59 PM, tartenon-at-netscape.net-at-sparc5.microscopy.com writes:
} My Understanding is that you can increase that resolution using Interpolation, } but the real resolution of the image will be 3.2 Megapixels with the Nikon } 995. I do not belive Olympus has 4 Megapixels non-interpolated (nor any } available digital camera)
It's really a lot more complicated than that. The actual spatial resolution of digital cameras is not simply the number of transistors on the chip (and in that regard I have the new Sony DSC-f707 which has 5 megapixels on the chip and produces awesome images, as compared to my Nikon 995). The problem is that the single chip cameras use an array of filters to expose some transistors to red, green and blue light. In order to get the image that is stored, they use interpolation to fill in the color information where it was not directly measured. The filters have broad wavelength coverage, and the interpolation schemes are pretty good (lots of patents in that area). But by direct measurement based on the Fourier power spectra none of these cameras has a spatial resolution that approaches the value you would expect based on the number of pixels in the stored image (which is usually the same as the number of transistors, except for Fuji who save images that have even more than that). For the Nikon and Sony cameras it is typical to find that the actual resolution elements across an image - beyond which you have empty magnification - number about 2/3 of the number of pixels. So a camera with 2000 transistors on each line (e.g., 2000 x 3000 = 6 megapixels) would probably measure as having about 1300-1350 elements of resolution (in other words, if you reduced the image to that size you would not really be discarding any information, and any enlargement beyond that size is empty).
The spatial resolution of the 5 M-pixel Sony is not as good as a 35 mm film camera, but it is extremely good all the same and certainly represents the best quality available now (and probably good enough for a great many applications) provided you store the image as uncompressed tif and don't throw away the important details by allowing the camera to use jpeg compression (which all of these cameras will do by default, to save memory). The tonal resolution - range of brightness values from dark to bright - is much more problematic. I have had good experience using high end consumer digital cameras for bright field microsopy, but they don't begin to have enough range for darkfield or fluorescence work. Remember that 8 bits is 256 grey levels, and even the cameras with internal 10 bits or more only produce about 8 bits on output because of the conversion from a linear detector to a film-like log output. Film easily covers several thousand discernible brightness steps, which would require a 12 bit or more range. That's why cooled cameras are used for these applications, and why the tiny chips used in consumer cameras won't work (the small transistors simply can't hold enough electrons to give that kind of dynamic range).
If you want to compare consumer type cameras, there is a wealth of unbiased information available online at {http://www.dpreview.com/reviews/specs.asp} . The discussion is centered on more typical photographic applications but there are comparative images, and a lot of info.
Digital cameras are great, and they save a lot of time and money. But don't expect an inexpensive consumer type camera to cover all applications.
Pixera 600-series digital camera produce non-interpolated honest/real 5.8M pixels, RGB.
gary g.
At 09:02 AM 2/7/2002, you wrote:
} Sorry You're Right I miss type the numbers it really is 3.2 Megapixels } where 2048 X 1536 = 3,145,728 (3.2 Megapixels) I'm using Nikon 995 and I } can Photograph Bright, Fluorescence Images My Understanding is that you } can increase that resolution using Interpolation, but the real resolution } of the image will be 3.2 Megapixels with the Nikon 995. I do not belive } Olympus has 4 Megapixels non-interpolated (nor any available digital camera)
Polaroid will not deliver the holder insert. Period. They did make a few of them. I saw a metal one, photocopied it and measured it. I have the insert spec's but not at home here. I could scan it and post it on my web page. The thing is only a thin but sturdy piece of sheet metal with rounded edges that has two slots to fit the two pegs in the 4X5 holder. The holder is bent or stamped to be slightly recessed, to equal the thickness of a sheet of film and the recess is the size of a sheet of film. This recess depth matches the rubberized surface of the regular 4X5 holder. You can make one from copper sheet stock or brass shim. You would make the outer thin and slotted piece the thickness of your film. Then solder on a thin piece of copper below the first piece to hold the film 'recessed'. I bought the copper material but then it did this:
} From a previous email: I have a Sprintscan 45i using binuscan as a driver 1.0.3 (wish I didn't). I could not get Polaroid to furnish any type of holder for 3.25 by 4 inch cut film, like SO-163. I put the film in sideways and use that. That raises questions about the focal plane of the film and it bending in the holder. These are minimal problems for me. However sometimes when the film is dried, it can be bent. So, I also have my own economy fix that you can build yourself.
Materials needed: 1 or 2 sheets of the cardboard that come with Kodak SO-163 film. An old exposed sheet or a new yellow sheet of SO-163 film. One clear sheet of developed clear and colorless SO-163 film. 20 inches of double back tape. A roll of 3M Scotchbrand® magic mending tape. One pair of scissors. Access to a paper cutter, optional. One 6 inch ruler with metric millimeters on it. One Sprintscan 45i 4X5 metal film holder. One magic marker to blacken all surfaces of the cardboard above (optional).
Cut one sheet of the cardboard so that it is square, 93 mm long and 36 mm high. It should fit snuggly in the opening of the film holder towards the locking nut or screw end. Once fitted, place a strip of 3M mending tape on the length of it so that only half of the tape is pressed onto the 93 mm long side by the opening for the film. Fold the tape over tightly, no wrinkles, and press the rest of the tape unto the opposite side of the blackened cardboard. This 93 mm edge will not fray from now on. Cut off any excess tape overhanging the ends. Take your colored film sheet and cut two pieces from the two good 4 inch straight edge sides. One piece will be "4 inches" ( actually 3 and 7/8ths) by 27 mm. The other will be "4 inches" by 12 mm. Take a piece of scrap 3 1/4 X 4" TEM film with an image on it and decide how much viewing area you are willing to sacrifice or lose at the edge of the micrograph that will go up against the cardboard side. 1-2 mm should be about right for a Philips CM12 piece of SO-163 3 ¼ X 4" cut film.
Place a 4 inch piece of double back tape on the back of the 27 mm wide piece so that it is flush against the 4 inch edge of it. Place it on the cardboard so that this 4 inch edge is recessed 1-2 mm from the 4 inch edge of the cardboard and centered so that 2-3 mm is overhanging the ends of the cardboard. Repeat this with the 12 mm piece and place it exactly on top of the first colored film piece. You now have a piece of cardboard with two pieces of centered colored film taped to it but inset 1-2 mm. The overhanging film edges will support this template holder in the Polaroid 4x5 holder, when installed.
(You can install more cardboard on the bottom of the first cardboard piece, if you want the holder to be more rigid.)
Take the 3.25 inch edge of the scrap colorless or developed film and cut a piece off 3 ¼ inches by 10 mm. Using double backed tape, install the tape so that it's length covers all 3.25 inches and it is set back 1-2 mm from the straight 3.25 inch edge of this colorless film piece. Turn it over, center it lengthwise over the dual stack of film and so that the colorless film's 3.25 inch edge is flush with the edge of the cardboard, not the colored dual film pieces. Press it into place on top of the colored film stack. You have now created a two film thick slot to insert your next TEM film into that you want to scan and it will be automatically aligned. The top piece is colorless to provide a view of how you are inserting the negative being mounted.
One job needs to be done. There is a peg on the 4x5 holder that your film to be scanned will rest up against. There is another one over by the locking mechanism. Cut the edge of the new template you made so that it has a notch that lines up with the peg nearest the locking screw. Press the ends of the template down onto the holder making sure it is square in the holder. You should now have an opening that is about 93 mm by 76 mm. That will be the area of your film that will be scanned. You will notice that the white or blackened cardboard is below the edge of the metal holder. Put a few pieces of mending tape around the template to hold it in place permanently and to keep it from sliding around.
To load the film holder: Slide the new film into the slot so that it is above the white cardboard and below the clear film piece. Slide the rest of the film up until it hits the one peg nearest the hinge. Square up the film if needed. Close the film holder. The film will be centered in the holder and flat all around the edges. The best part is this holder is FREE and only should take an hour or two to make.
My Polaroid scanner lasted 1½ years and is now defunct. When I turn it on, it moves the holder in and out continuously. It never initializes. It's not my favorite piece of equipment. I use a Powerlook III and it's much faster to scan with it. It does not have the OD range and resolution is worse but it didn't cost $7000. My customers don't complain. Use the Fuji setting for low constrast negs. Use regular transmission on high contrast negs. and invert.
Info on ordering UMAX scanner PLASTIC cut film holders that won't scratch the glass on your flat bed: Umax Phone: Area 510 - 651 - 4000 ext 3038 Parts are ordered from Fremont, Calf. only.
#SKIT-29002 PKG of 3 4" by 5" plastic cut film holders. $39.99
??? PKG of 5 2.25" by 3.25" holder $49.99 These holders can be routed out to make a 3 1/4 X 4" holder. It takes some skill to do this. Placing the 3x4 film in sideways in a 4x5 holder works just fine and that's what I use.
Paul Beauregard Sr. Research Associate PPG Industries Monroeville, PA 15146
} Last year I purchased this scanner and included a 3 1/4 x 4 1/4 negative } holder on the P.O., and have been waiting for the holder. Polaroid told } my sales rep that such a holder never existed and that John Warren no } longer worked a polaroid. } } } } Any help would be appreciated. } } Thanks, } } Don } } ______________________________________________________________________ } Donald L. Lovett e-mail: lovett-at-tcnj.edu } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } P.O. Box 7718 fax: (609) 637-5118 } The College of New Jersey } Ewing, NJ 08628-0718 } } } } } }
} Last year I purchased this scanner and included a 3 1/4 x 4 1/4 negative } holder on the P.O., and have been waiting for the holder. Polaroid told } my sales rep that such a holder never existed and that John Warren no } longer worked a polaroid. } } I have (and love) the Polaroid scanner, but am frustrated about not having } the appropriate film holder. Have any of you received this item, does it } have a part number, and how did you come to have it? } } Any help would be appreciated. } } Thanks, } } Don }
I assure all of you who responded to me that I will NOT be just regurgitating the information you have sent me. All the information was greatly appreciated, and hopefully I can understand it all to apply to my assignment. This is NOT just a homework assignment, and there is still much more that I am required to research and think about and discuss with my group before producing a final solution. The goal of our class is to learn how to research using different types of sources, and one of the ways we are told, when one has a limited amount of time to learn something, is to ask others who are more knowledgeable in the subject, professionals like yourselves. So thank you for your time for those who were kind enough to help me. Sincerely, Jenny
Quoting Michael O'Keefe {MAOKeefe-at-lbl.gov} :
} Don: } } I'm afraid I disagree with you and Tom. Part of a scientist's training } is to learn } how to identify and use resources. And we (collectively) are a resource } that I } believe should be made available to all who can benefit. Of course, } that leaves open } the question of whether the student benefits more by working things out } in isolation, } or by seeking guidance from experts and either really understanding the } answers or } (hopefully not) merely regurgitating them by rote. One hopes that the } intelligent } student will reject the latter course. } } Mike } } Donald Lovett 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 } } } -----------------------------------------------------------------------. } } } } Tom: } } } } I read Ms Wang's message, thought the same thoughts you articulated } here, } } and deleted the message. I think that we as a group should agree not } to } } do homework for students. Thanks for sharing your thoughts with us } and } } raising the issue. } } } } Don } } } } On Wed, 6 Feb 2002, Tom Phillips 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 think your instructor's hope would be that you figured out the } } } answers to class problems on your own. Asking an expert in the } field } } } and then simply regurgitating that information is a worthless } } } exercise. If you are going to invest the time and money required } to } } } earn a degree, you might want to try to learn something along the } } } way. I am a big supporter of listservers but hate to see them } used } } } in this way. } } } } } } } } } } } } } } } } } } To whom it may concern: } } } } } } } } I am an undergraduate student enrolled at the Georgia Institute } of } } } } Technology. } } } } I am in a biomedical engineering class, and we have a problem that } we must } } } } solve involving electron microscopes. I have a few questions that } I hope will } } } } get answered ASAP: } } } } } } } } 1. What are the advantages and disadvantages in using electrons } for } } } } microscopy } } } } rather than light? } } } } 2. Does the wavelength of the electrons have anythign to do with } the spatial } } } } resolution that the microscope produces in the final picture? } } } } 3. What is temporal resolution and how is it produced in the } electron } } } } microscope? } } } } } } } } Thank you for your time. I greatly appreciate your efforts in } helping me } } } } understand more of this subject. } } } } } } } } Sincerely, } } } } Jenny Wang } } } } } } } } ------------------------------------------------- } } } } Sent through Cyberbuzz- A Server for the Students } } } } http://cyberbuzz.gatech.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) } } } } } } } } } } } ______________________________________________________________________ } } Donald L. Lovett e-mail: lovett-at-tcnj.edu } } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } } P.O. Box 7718 fax: (609) 637-5118 } } The College of New Jersey } } Ewing, NJ 08628-0718 } }
------------------------------------------------- Sent through Cyberbuzz- A Server for the Students http://cyberbuzz.gatech.edu/
The deadline for abstract submission for the Microscopy & Microanalysis 2002 (Quebec City, Canada) is rapidly approaching. Among the featured Symposia at this year meeting, "Electron Microscopy of Macro-, Micro- and Meso-porous Materials" will address the current state of the art.
The deadline for electronic abstract submission is February 15th. We look forward to seeing you in Quebec City!
Manuel E. Brito, AIST, Japan Douglas Blum, ORNL, USA Dear Colleagues,
-- _/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/ National Institute of Advanced Industrial Science and Technology Synergy Materials Research Center
Manuel E. Brito, Eng. D.
Moriyama-ku, Nagoya 463-8687 JAPAN Tel: +81-52-739-0135 Fax: +81-52-739-0136 e-mail: manuel-brito-at-aist.go.jp
Hi, I think you have meant 2160 lines per mm giving 0.46 um between two adjacent lines (a standard replica grating); 2600 lines per mm should give .38 um spacing.
Best regard from Prague Oldrich
On 6 Feb 2002 at 15:30, Dusevich, Vladimir 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 } -----------------------------------------------------------------------. } } } } } } -----Original Message----- } } From: Brian Wajdyk [mailto:electronmicroscopist-at-hotmail.com] } } Sent: Wednesday, February 06, 2002 10:47 AM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: measurement and calibration onthe SEM } } } } } } -------------------------------------------------------------- } } ---------- } } 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 } } -------------------------------------------------------------- } } ---------. } } } } } } Fellow Microscopists, } } } } SEM has historically been used for metrology of various } } structures. I can't } } seem to find much literature about the artifacts associated } } with this type } } of measurement. We do use the NIST standards to check the } } calibration of } } our equipment, but I haven't characterized how the different } } beam or sample } } parameters effect the measurements. What do you do? Has } } anyone figured out } } his or her actual accuracy and precision? We have found that } } we can safely } } give measurements within +/-5% taking into account most human } } and equipment } } errors. This is based on the precision of measurements made of NIST } } structures, measured the same way, over several years. On } } the other hand, } } the smallest structure we can measure on the standard is 2um } } (line and } } space. How do you determine at what magnification you will no longer } } Cheap grating replicas with 2600 lines per mm give 0.46 um per line, } and this is not too bad for calibrating 50,000 magnification. I am } happy I do not need certified standards. } } } guarantee the measurement? I see that my MRS-3 from Geller } } says it's for } } 10x to 50kX. How do they figure out that the max magnification it is } } useful? Maybe it as simple as being able to fit the structure on the } } Maximum useful magnification is very specimen dependant, especially } for low voltage and low vacuum modes. Of course, for digital images } it is possible to check brightness profiles and if they have slopes } on edges of features, then measure "size" on half height of the slope. } But I am not aware about publications which dependably justify this kind } of measurements (manipulations with brightness and contrast and } specimen tilt could change slopes significantly). } } } screen. If that were true, you would expect that the } } instrument would also } } be calibrated to a much higher magnification. How high could } } I say it is } } accurate to? Can I safely measure a 1000A line assuming no } } It depends on resolution for your microscope/specimens and on } calibration standard you are using. And I think periodic lines with } spacing 2 um not really good standard to measure feature with } the size of 0.1 um. } } } obvious issues } } (i.e. drift)? Can anyone educate me more on this topic or } } If you have visible drift during single exposure, then something } wrong with microscope or specimen preparation technique. } } } point me to } } resources? } } } } } } Things that could effect measurements (feel free to add to list): } } Drift (mechanical / beam)- } } exposure time should be small for significant drift. } } } Charging (obvious or stretching of image from a slow scan) } } Magnification (adjusted for each set of lens relays) } } Could be eliminated with proper calibration. } } } kV (surface vs. subsurface image) } } Working Distance } } Could be eliminated with proper calibration. } } } Delineation method (raised vs. depression, materials contrast) } } Amount of delineation (3D effect) } } Resolution (near resolution limit of SEM?) } } Contrast (or lack of, bright / dark line) } } Edge effect (bright line) } } Consistency between tools (calibration, etc.) } } Consistency between methods (SEM, TEM, ellipsometry, AFM, etc.) } } Operator's eye (where to measure. Measure outside to inside, } } center to } } center, out to out, in to in?) } } Variance in measured layer thickness (topography, sloped } } profile (i.e. base } } larger than top)) } } Angle to beam } } Preparation methods (polish (i.e. smearing), cleave (i.e. } } pull of soft } } material), FIB (i.e. angle)) } } Type of algorithm if doing it automatically (i.e. %50 threshold) } } Some of the things you have mentioned relate to specimen/experiment, } to stereology, but not to microscope. For example, if I need to measure } size of depression without sharp edges, I have to find (or at least to } declare)right procedure for it's measurements. May be I have to perform } stereo measurements and define an edge as a place, where a depth of } depression become equal to 0.1 um (or 10% of total depth, or } whatever else, depending on a study). } } And thank you for your extensive list - it is very helpful for } observation of the problem. And about additions to your list - } I think everybody can say something. For example recently I tried } to measure in ESEM thickness of a layer which, as it turned out, } was a viscous liquid... } } Regards, } } Vladimir } } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } }
+-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of electron microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-2-4752399 Fax: +420-2-4752347 WEB:http://www.biomed.cas.cz/mbu/lem113/lem.htm
Greetings all; can anyone provide the formulation for Hank's buffer? I have looked in all of my books (most, but not all of which, admittedly, lean towards plants) and can't find a single reference. It is called for in a protocol for controlling autofluorescence in aldehyde fixed tissue.
thanks in advance shea
Dr. S. Shea Miller Agriculture & AgriFood Canada Eastern Cereal & Oilseed Research Centre Rm. 2068, K.W. Neatby Bldg. Central Experimental Farm Ottawa, Ontario, Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 E-mail: millers-at-em.agr.ca
I have two sets of data to interpreted: 1) X-ray rocking curve data, to analyze the defect (damage) of the wafer surface. 2) XPS data, to trace how the wafer surface composition will change with the increasing of sputtering time.
I need to have some fundamental idea of these two theories to interpreted data. Do you have any recommendations of the textbooks, or other papers I should read?
I have been quite happy with SIS Analysis package and database. We generate about 10-15k images a year here and it does a nice job managing it all and keeping relevant data with the images. My users may place a copy of the full program on their PC and run the analysis offline hooking directly to the network served db (MS Access based) and a new option will soon allow my users to log in with a web browser and query the db for their data directly- password protected and everything (hit a couple of snags and don't have it worked out yet). All three SEM's capture directly into the db, TEM data is dragged and dropped after capture, and I hope to soon have the LM directly capturing into it as well. I just haven't gotten it installed yet.
Hitachi distributes PCI quartz which seems very similar though I have no experience with it.
On the cheap side there is a program called thumbs+ from Cerious software for simple archiving.
Scott Whittaker SEM Lab Manager Smithsonian Institution PO Box 37012 MRC104 National Museum of Natural History Washington DC 20560-0104 202-357-1651
} } } "Comstock, Robert J." {comstorj-at-westinghouse.com} 02/07/02 11:42AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have progressed during the past few years to where the majority of our optical and SEM images are digital rather than Polaroid prints. In addition, we also scan TEM negatives and store them digitally. I am interested in some recommendations for software that can store the images in a database so they can be easily retrieved by keywords and also software for image analysis (e.g., particle size, image analysis, etc.) What options are available that people have experience with. I'd be interested in hearing both pro and con.
Thanks,
Bob Comstock Westinghouse Electric Co. Pittsburgh, PA 15235
I'm rather embarrassed that some of the members of this listserver were too narrow minded and/or arrogant to see that Ms. Wang was using this group of experts as a source, just as one would use a book. And besides, if she were just going to "regurgitate" the information learned here, wouldn't she just be doing the same with information pulled from a book?
I suddenly feel dirty somehow... Oh my God... It won't wash off!
Jeff (I'm in for it now) Oakley
} } I think your instructor's hope would be that you figured out the } } answers to class problems on your own. Asking an expert in the field } } and then simply regurgitating that information is a worthless } } exercise. If you are going to invest the time and money required to } } earn a degree, you might want to try to learn something along the } } way. I am a big supporter of listservers but hate to see them used } } in this way. } } } } } } } } To whom it may concern: } } } } } } I am an undergraduate student enrolled at the Georgia Institute of } } } Technology. } } } I am in a biomedical engineering class, and we have a problem that we must } } } solve involving electron microscopes. I have a few questions that I hope will } } } get answered ASAP: } } } } } } 1. What are the advantages and disadvantages in using electrons for } } } microscopy } } } rather than light? } } } 2. Does the wavelength of the electrons have anythign to do with the spatial } } } resolution that the microscope produces in the final picture? } } } 3. What is temporal resolution and how is it produced in the electron } } } microscope? } } } } } } Thank you for your time. I greatly appreciate your efforts in helping me } } } understand more of this subject. } } } } } } Sincerely, } } } Jenny Wang
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******************************************************************* Lucille A. Giannuzzi, Ph.D.
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by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id JAA17530 for dist-Microscopy; Fri, 8 Feb 2002 09:00:11 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id IAA17527 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 8 Feb 2002 08:59:40 -0600 (CST) Received: from www.matscieng.sunysb.edu (www.matscieng.sunysb.edu [129.49.36.33]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id IAA17520 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 8 Feb 2002 08:59:29 -0600 (CST) Received: (from jquinn-at-localhost) by www.matscieng.sunysb.edu (8.11.6/8.11.6) id g18ErCT04893 for Microscopy-at-sparc5.microscopy.com; Fri, 8 Feb 2002 09:53:12 -0500
Don -
College students should not broadcast messages seeking answers to elementary questions. Additionally, the use of "ASAP" was a bad idea.
JQuinn
PS: I sent Jenny Wang's message to her Chairperson and UG Program Director.
} From Microscopy-request-at-sparc5.microscopy.com Fri Feb 8 03:44:15 2002 } Date: Thu, 07 Feb 2002 13:47:36 -0800 } From: "Michael O'Keefe" {MAOKeefe-at-lbl.gov} } Organization: Lawrence Berkeley National Laboratory } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Questions on the Electron Microscope } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Don: } } I'm afraid I disagree with you and Tom. Part of a scientist's training is to learn } how to identify and use resources. And we (collectively) are a resource that I } believe should be made available to all who can benefit. Of course, that leaves open } } the question of whether the student benefits more by working things out in isolation, } } or by seeking guidance from experts and either really understanding the answers or } (hopefully not) merely regurgitating them by rote. One hopes that the intelligent } student will reject the latter course. } } Mike } } Donald Lovett 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 } } -----------------------------------------------------------------------. } } } } Tom: } } } } I read Ms Wang's message, thought the same thoughts you articulated here, } } and deleted the message. I think that we as a group should agree not to } } do homework for students. Thanks for sharing your thoughts with us and } } raising the issue. } } } } Don } } } } On Wed, 6 Feb 2002, Tom Phillips 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 think your instructor's hope would be that you figured out the } } } answers to class problems on your own. Asking an expert in the field } } } and then simply regurgitating that information is a worthless } } } exercise. If you are going to invest the time and money required to } } } earn a degree, you might want to try to learn something along the } } } way. I am a big supporter of listservers but hate to see them used } } } in this way. } } } } } } } } } } } } } } } } } } To whom it may concern: } } } } } } } } I am an undergraduate student enrolled at the Georgia Institute of } } } } Technology. } } } } I am in a biomedical engineering class, and we have a problem that we must } } } } solve involving electron microscopes. I have a few questions that I hope will } } } } get answered ASAP: } } } } } } } } 1. What are the advantages and disadvantages in using electrons for } } } } microscopy } } } } rather than light? } } } } 2. Does the wavelength of the electrons have anythign to do with the spatial } } } } resolution that the microscope produces in the final picture? } } } } 3. What is temporal resolution and how is it produced in the electron } } } } microscope? } } } } } } } } Thank you for your time. I greatly appreciate your efforts in helping me } } } } understand more of this subject. } } } } } } } } Sincerely, } } } } Jenny Wang } } } } } } } } ------------------------------------------------- } } } } Sent through Cyberbuzz- A Server for the Students } } } } http://cyberbuzz.gatech.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) } } } } } } } } } } ______________________________________________________________________ } } Donald L. Lovett e-mail: lovett-at-tcnj.edu } } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } } P.O. Box 7718 fax: (609) 637-5118 } } The College of New Jersey } } Ewing, NJ 08628-0718 } }
In a message dated 2/8/02 10:07:03 AM, jquinn-at-www.matscieng.sunysb.edu writes:
} College students should not broadcast messages seeking answers } to elementary questions.
Some of them do things even worse than that. As the author of a moderately well known book on image analysis I get several messages each week asking questions that boil down to something like this:
"I've been asked to report on X. Could you give me a concise answer so I won't have to read and digest all of the information in your text? Oh, and I need it by tomorrow.
The only question that is even more annoying is "I can't afford your book. Would you please send me a copy?"
The art of reading, digesting and combining information from multiple sources is vital in education. To try to short cut this and get someone else to chew the food for you and then regurgitate it is not only lazy and dishonest, it also prevents students from learning to think, which is a more important part of education than the factual stuff they seem to be dealing with.
While Jenny's questions were rather vague and sounded like she was looking for an easy answer to an assignment, we should probably give students the benefit of the doubt. How many times have *we* presented ill-formed questions when we were not quite sure of what we were asking?
I agree that we should not do a student's assignment for him, but perhaps we can somewhat more gently steer them to the source of the answers rather than flame them. I think that if the questions sound inappropriate, we can make a comment to the effect that we're not going to provide the answer, but only the source of the info. The ensuing discussion may lead to a sharpening of the question as the student thinks though what he is trying to ask. While there are, indeed, students looking for the easy way out, we need to be careful not to flame the student who framed the question poorly.
Cheers, Henk Colijn
{...much deleted material...}
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Fools are pleased when they discover error. The wise are pleased when they discover truth.
Jeff: I am assuming from your rayovac.com you are not an educator. That doesn't disqualify your opinion but I would be more worried if other currently teaching academics widely expressed this view. Learning to look something up in the library is part of the teaching assignment. Finding the correct information and distilling it is not trivial. Most experts on the listserver could answer each of those questions in a few concise sentences. You would be hard pressed to find any source in a library in which you found the question followed by the answer. When you read the literature, lots of time you end up reading additional information on peripheral topics that add to the learning process. More importantly, students constantly give me sentences in their papers that are clearly paraphrased from the literature. I am not suggesting this constitutes plagiarism but it is often apparent from the sentence that they don't really understand what it means. They think they do but when I discuss it with them, they are unable to explain the sophisticated sentence in basic terms. Students frequently comment in my teaching evaluations that the most important thing they learned in class was that memorizing and rephrasing the literature doesn't equate to real understanding. If the instructor wanted them simply to ask an expert to get the bottom line answer, why didn't the instructor simply say it in lecture or give them a handout? Don't you think the instructor knew? Do you think a bioengineering class was designed to teach web surfing tools. I don't know if you should feel dirty but I think your batteries need recharging. Tom Phillips
} } } I'm rather embarrassed that some of the members of this listserver were too } narrow minded and/or arrogant to see that Ms. Wang was using this group of } experts as a source, just as one would use a book. And besides, if she were } just going to "regurgitate" the information learned here, wouldn't she just } be doing the same with information pulled from a book? } } I suddenly feel dirty somehow... Oh my God... It won't wash off! } } } Jeff (I'm in for it now) Oakley } } } } } } I think your instructor's hope would be that you figured out the } } } answers to class problems on your own. Asking an expert in the field } } } and then simply regurgitating that information is a worthless } } } exercise. If you are going to invest the time and money required to } } } earn a degree, you might want to try to learn something along the } } } way. I am a big supporter of listservers but hate to see them used } } } in this way. } } } } } } } } } } } To whom it may concern: } } } } } } } } I am an undergraduate student enrolled at the Georgia Institute of } } } } Technology. } } } } I am in a biomedical engineering class, and we have a problem that we } must } } } } solve involving electron microscopes. I have a few questions that I } hope will } } } } get answered ASAP: } } } } } } } } 1. What are the advantages and disadvantages in using electrons for } } } } microscopy } } } } rather than light? } } } } 2. Does the wavelength of the electrons have anythign to do with the } spatial } } } } resolution that the microscope produces in the final picture? } } } } 3. What is temporal resolution and how is it produced in the electron } } } } microscope? } } } } } } } } Thank you for your time. I greatly appreciate your efforts in helping } me } } } } understand more of this subject. } } } } } } } } Sincerely, } } } } Jenny Wang
-- 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)
Listers, I am in need of a used but functional EDS detector ported for an ETEC AUTOSCAN SEM. Preferences being Tracor/Noran, Kevex, PGT or EDAX, T/W if possible. If anyone has any leads, please feel free to contact me off line. Thanks in advance.
Gary M. Easton, President Scanners Corporation 90 Aileron Court, Suite 6 Westminster, Maryland USA 21157 410.857.7633(v) 410.857.7636(f) www.scannerscorp.com
Which software environment you choose depends largely on your needs. Most people need a standard package which is easy to use and there are several software packages on the market. Do you want to do basic image analysis or do you ever want to do more complicated analysis on large datasets, this will influence your choice. Most users "use" imaging software and don't do a lot of basic image algorithm development themselves.
Most of the software is available for both Windows and Macintosh, like the "NIH-Image" family. There is also AnalySIS and ImagePro Plus. For EM there is specialised software from Gatan and a software package like KHOROS is also suitable.
There are several image databases available, I believe there are now several packages available with a web based interface which enables you to browse the database through a webbrowser.
Best regards,
Peter
} -----Original Message----- } } From: Comstock, Robert J. [mailto:comstorj-at-westinghouse.com] } Sent: Thursday, February 07, 2002 11:43 AM } To: MicroscopyListserver (E-mail) } Subject: Database/image analysis for digital images } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } We have progressed during the past few years to where the majority of our } optical and SEM images are digital rather than Polaroid prints. In addition, } we also scan TEM negatives and store them digitally. I am interested in } some recommendations for software that can store the images in a database so } they can be easily retrieved by keywords and also software for image } analysis (e.g., particle size, image analysis, etc.) What options are } available that people have experience with. I'd be interested in hearing } both pro and con. } } Thanks, } } Bob Comstock } Westinghouse Electric Co. } Pittsburgh, PA 15235
-- Dr. Peter Van Osta
Union Biometrica N.V./S.A. European Scientific Operations
I am new to this listserve so I hesitated to respond, but here it goes. I think a student's questions should be answered in a fashion as you suggest. Mining all resources for information (including media such as this) is a very necessary skill in todays (and definitely tomorrows) world. Whether we agree or not, searching table of contents in the hardcopy library is becoming less and less valuable. Having high school students as children, I have been exposed to a tremendous opportunity for expeditious research covering a broad spectrum of resources using this media. At this time, a combination of hardcopy library and electronic media seems appropriate.
The concern of regurgitation may be moot. After all, from what I have read, this student may very well have the best of intentions and will list this server as her information source. This is an ethical question only she can answer for herself. In addition, it's certainly possible that this is a small fraction of her group's assignment and gleaning the answers to preliminary questions here will only open doors to deeper understanding later.
Obviously, to use a resource such as this to do frequent homework assignments is a mis-use of our time. However, the natural and logical consequences are for the student to deal with when he/she reconciles with the ethical questions and attempts to enter the job market.
Jenny, you are to be commended on your fine response to the negative comments of some of "my" listserver colleagues. Those of us with 10-25 years of "hand's on experience" in various aspects of microscopy are a valuable asset of knowledge for new students in our field. Likewise, there is much we can hopefully learn your age group. Good luck!
Bruce F. Ingber, Biologist USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124 bingber-at-srrc.ars.usda.gov 504-286-4270 phone 504-286-4419 fax
We need to purchase a new calibration standard for our SEM that gets us down to the next level. We need to have accuracy down to 0.010 microns. We have been using the Geller MRS-3 and now are considering purchasing the MRS-4 traceable standard. I believe this will give us what we need but I was wondering if there are other standards out there that are better, the same, worse? I need to hear from the calibration specialists out there. Thanks,
Mark Windland Honeywell Minneapolis, Minnesota 763-954-2845
I am looking for some SEM images of semiconductor circuits. Would like to have a range of magnifications and view angles from those that show whole die with bond pads and leads to close-ups of circiut elements. Looking for interesting features and topography. These will be used as guides to build some 3D models for a SEM animation video I am working on. If anyone can supply images for this project please send them to me via E-mail.
Thanks for your help!
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
Though I was not in on the discussion, I am going to throw in my two cents.
When I read the original post, it looked like someone had a test sheet, or assignment sheet, with those questions on it. The request appeared to be for direct answers to those questions. It did not surprise me that this fine collection of knowledge would conclude that someone was trying to shortcut the leaning process, and avoid really learning a subject. If the student was going to just "regurgitate" something from a book, then we have not contributed to the current decay in the quality of the knowledge, and understanding, in the students receiving degrees. Those posts that did give excellent research sources, ignoring the APPARENT shortcut, did well in my view. If the request was for help in understanding a concept, function, process, etc., then a more direct answer from an expert becomes extremely valuable.
I don't think ANYBODY on the list should be embarrassed or ashamed. Had Ms. Wang requested help in a different manner, she would have gotten a stream of replies with all sorts of information, and the simple, fill out the test, answers probably would have been in there, also.
Darrell
"Oakley, Jeff" {oakleyj-at-rayovac.com} on 02/08/2002 08:42:30 AM
To: Microscopy-at-sparc5.microscopy.com cc:
I'm rather embarrassed that some of the members of this listserver were too narrow minded and/or arrogant to see that Ms. Wang was using this group of experts as a source, just as one would use a book. And besides, if she were just going to "regurgitate" the information learned here, wouldn't she just be doing the same with information pulled from a book?
I suddenly feel dirty somehow... Oh my God... It won't wash off!
Jeff (I'm in for it now) Oakley
} } I think your instructor's hope would be that you figured out the } } answers to class problems on your own. Asking an expert in the field } } and then simply regurgitating that information is a worthless } } exercise. If you are going to invest the time and money required to } } earn a degree, you might want to try to learn something along the } } way. I am a big supporter of listservers but hate to see them used } } in this way. } } } } } } } } To whom it may concern: } } } } } } I am an undergraduate student enrolled at the Georgia Institute of } } } Technology. } } } I am in a biomedical engineering class, and we have a problem that we must } } } solve involving electron microscopes. I have a few questions that I hope will } } } get answered ASAP: } } } } } } 1. What are the advantages and disadvantages in using electrons for } } } microscopy } } } rather than light? } } } 2. Does the wavelength of the electrons have anythign to do with the spatial } } } resolution that the microscope produces in the final picture? } } } 3. What is temporal resolution and how is it produced in the electron } } } microscope? } } } } } } Thank you for your time. I greatly appreciate your efforts in helping me } } } understand more of this subject. } } } } } } Sincerely, } } } Jenny Wang
} Gentlemen, } } I disagree and find this attitude somewhat surprising coming from educators. } I took the time to answer her questions and encourage her in her studies. I } believe this list is just as valuable a resource as any other method she } could have used. I believe students should feel comfortable in using it in } the attempt to "learn something along the way".
Her instructor wanted her to research the answer, otherwise he would have told her the answers (to her very elementary questions) himself. Asking someone other than her instructor is not research. The way her questions were asked indicated to me that she was merely repeating questions she had been asked. As for identifying sources of information, a college-level biology text would have the answers she wanted. A text book on EM would have the answers. Even a web site on EM would have the answers. She did not look for any of those, she tried to get the answers handed to her with no more effort than a e-mail. She probable still does not know that all of those other resources exist. What will she do if her server goes down? At my institution, we insist that students look up simple, straightforward facts for themselves rather than using the faculty as encyclopeidas. That is what the textbook is for. Once they are in possession of the facts, we then ask them to use them to solve problems. We don't view reciting facts to students as higher education.
} Roy Beavers } Southern Methodist University } Dept. of Geological Sciences } Electron Microprobe Lab } P.O. Box 750395 } Dallas, Tx 75275 } voice: 214-768-2756 } fax: 214-768-2701 } E-mail: rbeavers-at-mail.smu.edu } } } } -----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-UMDNJ.EDU] } Sent: Thursday, February 07, 2002 11:59 AM } To: Donald Lovett } Cc: Tom Phillips; Microscopy-at-sparc5.microscopy.com } Subject: Re: Questions on the Electron Microscope } } ------------------------------------------------------------------------ } 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 agree with Don and Tom. } } Donald Lovett 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 } } -----------------------------------------------------------------------. } } } } Tom: } } } } I read Ms Wang's message, thought the same thoughts you articulated here, } } and deleted the message. I think that we as a group should agree not to } } do homework for students. Thanks for sharing your thoughts with us and } } raising the issue. } } } } Don } } } } On Wed, 6 Feb 2002, Tom Phillips wrote: } } } } } } } I think your instructor's hope would be that you figured out the } } } answers to class problems on your own. Asking an expert in the field } } } and then simply regurgitating that information is a worthless } } } exercise. If you are going to invest the time and money required to } } } earn a degree, you might want to try to learn something along the } } } way. I am a big supporter of listservers but hate to see them used } } } in this way. } } } } } } } } } } } To whom it may concern: } } } } } } } } I am an undergraduate student enrolled at the Georgia Institute of } } } } Technology. } } } } I am in a biomedical engineering class, and we have a problem that we } must } } } } solve involving electron microscopes. I have a few questions that I } hope will } } } } get answered ASAP: } } } } } } } } 1. What are the advantages and disadvantages in using electrons for } } } } microscopy } } } } rather than light? } } } } 2. Does the wavelength of the electrons have anythign to do with the } spatial } } } } resolution that the microscope produces in the final picture? } } } } 3. What is temporal resolution and how is it produced in the electron } } } } microscope? } } } } } } } } Thank you for your time. I greatly appreciate your efforts in helping } me } } } } understand more of this subject. } } } } } } } } Sincerely, } } } } Jenny Wang } } } } } } } } ------------------------------------------------- } } } } Sent through Cyberbuzz- A Server for the Students } } } } http://cyberbuzz.gatech.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) } } } } } } } } } } ______________________________________________________________________ } } Donald L. Lovett e-mail: lovett-at-tcnj.edu } } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } } P.O. Box 7718 fax: (609) 637-5118 } } The College of New Jersey } } Ewing, NJ 08628-0718 } } 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 } **********************************************
-- ********************************************** 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 **********************************************
John; You are correct about the small CCDs of consumer digital cameras have sever performance deficiencies due to their small pixel size. The F707 and other consumer digital cameras that use the same chip (2/3"-Nikon 5000 and Olympus E20N) suffer from noise even in visible light photographs. The Nikon D1X and the new Canon EOS 1D(advertised 11.5 µm pixels) are significant improvements, but cost $6000 just for the camera body! One organization addressing this problem is http://www.fredmiranda.com/Action_profilesPage/index.html where they discuss and sell Photoshop plug-ins for noise reduction. Another digital camera site I really like is: http://www.imaging-resource.com/
John Mardinly Intel
-----Original Message----- } From: "DrJohnRuss%aol.com"-at-sparc5.microscopy.com [mailto:"DrJohnRuss%aol.com"-at-sparc5.microscopy.com] Sent: Thursday, February 07, 2002 4:50 PM To: microscopy-at-sparc5.microscopy.com
In a message dated 2/7/02 12:22:59 PM, tartenon-at-netscape.net-at-sparc5.microscopy.com writes:
} My Understanding is that you can increase that resolution using Interpolation, } but the real resolution of the image will be 3.2 Megapixels with the Nikon } 995. I do not belive Olympus has 4 Megapixels non-interpolated (nor any } available digital camera)
It's really a lot more complicated than that. The actual spatial resolution of digital cameras is not simply the number of transistors on the chip (and in that regard I have the new Sony DSC-f707 which has 5 megapixels on the chip and produces awesome images, as compared to my Nikon 995). The problem is that the single chip cameras use an array of filters to expose some transistors to red, green and blue light. In order to get the image that is stored, they use interpolation to fill in the color information where it was
not directly measured. The filters have broad wavelength coverage, and the interpolation schemes are pretty good (lots of patents in that area). But by
direct measurement based on the Fourier power spectra none of these cameras has a spatial resolution that approaches the value you would expect based on
the number of pixels in the stored image (which is usually the same as the number of transistors, except for Fuji who save images that have even more than that). For the Nikon and Sony cameras it is typical to find that the actual resolution elements across an image - beyond which you have empty magnification - number about 2/3 of the number of pixels. So a camera with 2000 transistors on each line (e.g., 2000 x 3000 = 6 megapixels) would probably measure as having about 1300-1350 elements of resolution (in other words, if you reduced the image to that size you would not really be discarding any information, and any enlargement beyond that size is empty).
The spatial resolution of the 5 M-pixel Sony is not as good as a 35 mm film camera, but it is extremely good all the same and certainly represents the best quality available now (and probably good enough for a great many applications) provided you store the image as uncompressed tif and don't throw away the important details by allowing the camera to use jpeg compression (which all of these cameras will do by default, to save memory).
The tonal resolution - range of brightness values from dark to bright - is much more problematic. I have had good experience using high end consumer digital cameras for bright field microsopy, but they don't begin to have enough range for darkfield or fluorescence work. Remember that 8 bits is 256
grey levels, and even the cameras with internal 10 bits or more only produce
about 8 bits on output because of the conversion from a linear detector to a
film-like log output. Film easily covers several thousand discernible brightness steps, which would require a 12 bit or more range. That's why cooled cameras are used for these applications, and why the tiny chips used in consumer cameras won't work (the small transistors simply can't hold enough electrons to give that kind of dynamic range).
If you want to compare consumer type cameras, there is a wealth of unbiased information available online at {http://www.dpreview.com/reviews/specs.asp} .
The discussion is centered on more typical photographic applications but there are comparative images, and a lot of info.
Digital cameras are great, and they save a lot of time and money. But don't expect an inexpensive consumer type camera to cover all applications.
Having read the comments on both sides I tend to think that part of the problem was due to quite a bit of misunderstanding (mainly on our part). I too had the same negative reaction when I first read Ms. Wang's e-mail. The way the first e-mail read I felt that the student was trying to have others solve her assignment problem ( which is why I did not respond to her request). My reaction changed however when I read her latest e-mail explaining more the nature of the exercise. I hope I learned from this experience and that in the future I will have a better attitude and ask first for more information before I decide to give an answer.
Jordi
-----Original Message----- } From: Oakley, Jeff [mailto:oakleyj-at-rayovac.com] Sent: Friday, February 08, 2002 8:43 AM To: Microscopy-at-sparc5.microscopy.com
I'm rather embarrassed that some of the members of this listserver were too narrow minded and/or arrogant to see that Ms. Wang was using this group of experts as a source, just as one would use a book. And besides, if she were just going to "regurgitate" the information learned here, wouldn't she just be doing the same with information pulled from a book?
I suddenly feel dirty somehow... Oh my God... It won't wash off!
Jeff (I'm in for it now) Oakley
} } I think your instructor's hope would be that you figured out the } } answers to class problems on your own. Asking an expert in the field } } and then simply regurgitating that information is a worthless } } exercise. If you are going to invest the time and money required to } } earn a degree, you might want to try to learn something along the } } way. I am a big supporter of listservers but hate to see them used } } in this way. } } } } } } } } To whom it may concern: } } } } } } I am an undergraduate student enrolled at the Georgia Institute of } } } Technology. } } } I am in a biomedical engineering class, and we have a problem that we must } } } solve involving electron microscopes. I have a few questions that I hope will } } } get answered ASAP: } } } } } } 1. What are the advantages and disadvantages in using electrons for } } } microscopy } } } rather than light? } } } 2. Does the wavelength of the electrons have anythign to do with the spatial } } } resolution that the microscope produces in the final picture? } } } 3. What is temporal resolution and how is it produced in the electron } } } microscope? } } } } } } Thank you for your time. I greatly appreciate your efforts in helping me } } } understand more of this subject. } } } } } } Sincerely, } } } Jenny Wang
} ------------------------------------------------------------------------ } 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'm rather embarrassed that some of the members of this listserver were too } narrow minded and/or arrogant to see that Ms. Wang was using this group of } experts as a source, just as one would use a book.
Thank you. Do all disagreements with your educational philosophy fall under this umbrella?
} And besides, if she were } just going to "regurgitate" the information learned here, wouldn't she just } be doing the same with information pulled from a book?
The point is, she didn't use a book. Rather than look it up for herself she tried to get someone else to give her the answers. Rather like the old rubric about teaching a man to fish instead of giving him a fish.
} I suddenly feel dirty somehow... Oh my God... It won't wash off! } } Jeff (I'm in for it now) Oakley } } } } I think your instructor's hope would be that you figured out the } } } answers to class problems on your own. Asking an expert in the field } } } and then simply regurgitating that information is a worthless } } } exercise. If you are going to invest the time and money required to } } } earn a degree, you might want to try to learn something along the } } } way. I am a big supporter of listservers but hate to see them used } } } in this way. } } } } } } } } } } } To whom it may concern: } } } } } } } } I am an undergraduate student enrolled at the Georgia Institute of } } } } Technology. } } } } I am in a biomedical engineering class, and we have a problem that we } must } } } } solve involving electron microscopes. I have a few questions that I } hope will } } } } get answered ASAP: } } } } } } } } 1. What are the advantages and disadvantages in using electrons for } } } } microscopy } } } } rather than light? } } } } 2. Does the wavelength of the electrons have anythign to do with the } spatial } } } } resolution that the microscope produces in the final picture? } } } } 3. What is temporal resolution and how is it produced in the electron } } } } microscope? } } } } } } } } Thank you for your time. I greatly appreciate your efforts in helping } me } } } } understand more of this subject. } } } } } } } } Sincerely, } } } } Jenny Wang
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 **********************************************
You are correct, I am not an educator. I also agree with you whole heartedly that when someone reads a text in search of information, they learn a great deal more than they had planned on... It happens to me every time I pick up a book in search of an answer to a question. I'm sure this probably was the intent of the instructor.
The point I should have made in my previous post is that instead of shutting someone down and "scolding" them (which is exactly what some of the listers did) for what appeared to be a Cliff's Notes research method, the person should have been guided to useful web pages or texts that would have made them find the answers for themselves (which other posters did - kudos to those).
It is possible to be helpful while at the same time not giving someone a free ride.
I don't think my batteries are dead, Tom, I just think we are operating at different voltages.
Jeff
-----Original Message----- } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] Sent: Friday, February 08, 2002 9:26 AM To: Oakley, Jeff Cc: Microscopy-at-msa.microscopy.com
While I am no longer in academia, were that not the case, I might have a different view. However, consider the future relative to all students.
If they are aspiring to science, microscopy, etc., given their experience with MSA as a student, what might their opinion be when they become professionals? Isn't it possible that they could have a bad taste in their mouth about MSA in particular and the list specifically?
I don't belive that professionals should answer student's questions in a concise and packaged format. Rather, professionals should be used primarily as pointers to sources of information. Sometimes, they might provide specific data. Either way, it should (emphasis) stimulate the student towards their goal. If their motivation for seeking information from professionals is simply to get their assignment done, this could lead to several consequences. One of these is that they will not know the material and will be unable to perform as an employee.
It seems that this point is what most listers should be concerned about--and probably are.
} -----Original Message----- } From: Oldrich Benada [mailto:benada-at-biomed.cas.cz] } Sent: Friday, February 08, 2002 2:20 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: RE: measurement and calibration onthe SEM } } } -------------------------------------------------------------- } ---------- } 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, } I think you have meant 2160 lines per mm giving 0.46 um between two } adjacent lines (a standard replica grating); 2600 lines per mm should } give .38 um spacing. } } Best regard from Prague } Oldrich } } } } On 6 Feb 2002 at 15:30, Dusevich, Vladimir 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 } } } -------------------------------------------------------------- } ---------. } } } } } } } } } } } -----Original Message----- } } } From: Brian Wajdyk [mailto:electronmicroscopist-at-hotmail.com] } } } Sent: Wednesday, February 06, 2002 10:47 AM } } } To: Microscopy-at-sparc5.microscopy.com } } } Subject: measurement and calibration onthe SEM } } } } } } } } } -------------------------------------------------------------- } } } ---------- } } } 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 } } } -------------------------------------------------------------- } } } ---------. } } } } } } } } } Fellow Microscopists, } } } } } } SEM has historically been used for metrology of various } } } structures. I can't } } } seem to find much literature about the artifacts associated } } } with this type } } } of measurement. We do use the NIST standards to check the } } } calibration of } } } our equipment, but I haven't characterized how the different } } } beam or sample } } } parameters effect the measurements. What do you do? Has } } } anyone figured out } } } his or her actual accuracy and precision? We have found that } } } we can safely } } } give measurements within +/-5% taking into account most human } } } and equipment } } } errors. This is based on the precision of measurements } made of NIST } } } structures, measured the same way, over several years. On } } } the other hand, } } } the smallest structure we can measure on the standard is 2um } } } (line and } } } space. How do you determine at what magnification you } will no longer } } } } Cheap grating replicas with 2600 lines per mm give 0.46 um per line, } } and this is not too bad for calibrating 50,000 magnification. I am } } happy I do not need certified standards. } } } } } guarantee the measurement? I see that my MRS-3 from Geller } } } says it's for } } } 10x to 50kX. How do they figure out that the max } magnification it is } } } useful? Maybe it as simple as being able to fit the } structure on the } } } } Maximum useful magnification is very specimen dependant, especially } } for low voltage and low vacuum modes. Of course, for digital images } } it is possible to check brightness profiles and if they have slopes } } on edges of features, then measure "size" on half height of } the slope. } } But I am not aware about publications which dependably } justify this kind } } of measurements (manipulations with brightness and contrast and } } specimen tilt could change slopes significantly). } } } } } screen. If that were true, you would expect that the } } } instrument would also } } } be calibrated to a much higher magnification. How high could } } } I say it is } } } accurate to? Can I safely measure a 1000A line assuming no } } } } It depends on resolution for your microscope/specimens and on } } calibration standard you are using. And I think periodic lines with } } spacing 2 um not really good standard to measure feature with } } the size of 0.1 um. } } } } } obvious issues } } } (i.e. drift)? Can anyone educate me more on this topic or } } } } If you have visible drift during single exposure, then something } } wrong with microscope or specimen preparation technique. } } } } } point me to } } } resources? } } } } } } } } } Things that could effect measurements (feel free to add to list): } } } Drift (mechanical / beam)- } } } } exposure time should be small for significant drift. } } } } } Charging (obvious or stretching of image from a slow scan) } } } Magnification (adjusted for each set of lens relays) } } } } Could be eliminated with proper calibration. } } } } } kV (surface vs. subsurface image) } } } Working Distance } } } } Could be eliminated with proper calibration. } } } } } Delineation method (raised vs. depression, materials contrast) } } } Amount of delineation (3D effect) } } } Resolution (near resolution limit of SEM?) } } } Contrast (or lack of, bright / dark line) } } } Edge effect (bright line) } } } Consistency between tools (calibration, etc.) } } } Consistency between methods (SEM, TEM, ellipsometry, AFM, etc.) } } } Operator's eye (where to measure. Measure outside to inside, } } } center to } } } center, out to out, in to in?) } } } Variance in measured layer thickness (topography, sloped } } } profile (i.e. base } } } larger than top)) } } } Angle to beam } } } Preparation methods (polish (i.e. smearing), cleave (i.e. } } } pull of soft } } } material), FIB (i.e. angle)) } } } Type of algorithm if doing it automatically (i.e. %50 threshold) } } } } Some of the things you have mentioned relate to specimen/experiment, } } to stereology, but not to microscope. For example, if I } need to measure } } size of depression without sharp edges, I have to find (or } at least to } } declare)right procedure for it's measurements. May be I } have to perform } } stereo measurements and define an edge as a place, where a depth of } } depression become equal to 0.1 um (or 10% of total depth, or } } whatever else, depending on a study). } } } } And thank you for your extensive list - it is very helpful for } } observation of the problem. And about additions to your list - } } I think everybody can say something. For example recently I tried } } to measure in ESEM thickness of a layer which, as it turned out, } } was a viscous liquid... } } } } Regards, } } } } Vladimir } } } } } } Vladimir M. Dusevich, Ph.D. } } Electron Microscope Lab Manager } } 3127 School of Dentistry } } 650 E. 25th Street } } Kansas City, MO 64108-2784 } } } } Phone: (816) 235-2072 } } Fax: (816) 235-5524 } } Web: http://www.umkc.edu/dentistry/microscopy } } } } } } } } } +-----------------------------------+ } Oldrich Benada } Acad. Sci. CR } Institute of Microbiology } Laboratory of electron microscopy } Videnska 1083 } CZ - 142 20 Prague 4 - Krc } Czech Republic } +------------------------------------+ } Phone: +420-2-4752399 } Fax: +420-2-4752347 } WEB:http://www.biomed.cas.cz/mbu/lem113/lem.htm } } }
No offence Jeff, but does anyone else think that this thread has taken on some of the aspects of the Energizer Bunny? ;-)
"Oakley, Jeff" 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 } -----------------------------------------------------------------------. } } Tom, } } You are correct, I am not an educator. I also agree with you whole } heartedly that when someone reads a text in search of information, they } learn a great deal more than they had planned on... It happens to me every } time I pick up a book in search of an answer to a question. I'm sure this } probably was the intent of the instructor. } } The point I should have made in my previous post is that instead of shutting } someone down and "scolding" them (which is exactly what some of the listers } did) for what appeared to be a Cliff's Notes research method, the person } should have been guided to useful web pages or texts that would have made } them find the answers for themselves (which other posters did - kudos to } those). } } It is possible to be helpful while at the same time not giving someone a } free ride. } } I don't think my batteries are dead, Tom, I just think we are operating at } different voltages. } } Jeff } } -----Original Message----- } } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] } Sent: Friday, February 08, 2002 9:26 AM } To: Oakley, Jeff } Cc: Microscopy-at-msa.microscopy.com } Subject: RE: Questions on the Electron Microscope } } Jeff: I am assuming from your rayovac.com you are not an educator. } That doesn't disqualify your opinion but I would be more worried if } other currently teaching academics widely expressed this view. } Learning to look something up in the library is part of the teaching } assignment. Finding the correct information and distilling it is not } trivial. Most experts on the listserver could answer each of those } questions in a few concise sentences. You would be hard pressed to } find any source in a library in which you found the question followed } by the answer. When you read the literature, lots of time you end } up reading additional information on peripheral topics that add to } the learning process. More importantly, students constantly give me } sentences in their papers that are clearly paraphrased from the } literature. I am not suggesting this constitutes plagiarism but it } is often apparent from the sentence that they don't really understand } what it means. They think they do but when I discuss it with them, } they are unable to explain the sophisticated sentence in basic terms. } Students frequently comment in my teaching evaluations that the most } important thing they learned in class was that memorizing and } rephrasing the literature doesn't equate to real understanding. If } the instructor wanted them simply to ask an expert to get the bottom } line answer, why didn't the instructor simply say it in lecture or } give them a handout? Don't you think the instructor knew? Do you } think a bioengineering class was designed to teach web surfing tools. } I don't know if you should feel dirty but I think your batteries need } recharging. Tom Phillips } } } } } } } I'm rather embarrassed that some of the members of this listserver were too } } narrow minded and/or arrogant to see that Ms. Wang was using this group of } } experts as a source, just as one would use a book. And besides, if she } were } } just going to "regurgitate" the information learned here, wouldn't she just } } be doing the same with information pulled from a book? } } } } I suddenly feel dirty somehow... Oh my God... It won't wash off! } } } } } } Jeff (I'm in for it now) Oakley } } } } } } } } } } I think your instructor's hope would be that you figured out the } } } } answers to class problems on your own. Asking an expert in the field } } } } and then simply regurgitating that information is a worthless } } } } exercise. If you are going to invest the time and money required to } } } } earn a degree, you might want to try to learn something along the } } } } way. I am a big supporter of listservers but hate to see them used } } } } in this way. } } } } } } } } } } } } } } To whom it may concern: } } } } } } } } } } I am an undergraduate student enrolled at the Georgia Institute of } } } } } Technology. } } } } } I am in a biomedical engineering class, and we have a problem that we } } must } } } } } solve involving electron microscopes. I have a few questions that I } } hope will } } } } } get answered ASAP: } } } } } } } } } } 1. What are the advantages and disadvantages in using electrons for } } } } } microscopy } } } } } rather than light? } } } } } 2. Does the wavelength of the electrons have anythign to do with the } } spatial } } } } } resolution that the microscope produces in the final picture? } } } } } 3. What is temporal resolution and how is it produced in the } electron } } } } } microscope? } } } } } } } } } } Thank you for your time. I greatly appreciate your efforts in } helping } } me } } } } } understand more of this subject. } } } } } } } } } } Sincerely, } } } } } Jenny Wang } } -- } 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)
Sending Jenny Wang's message to her Chairperson and UG Program Director was, in my eyes, really unnecessary. Maybe it was just a nice try from Jenny Wang to get her homework done, but this is really overdone.
Regards,
Stefan
Jim Quinn wrote:
} Don - } } College students should not broadcast messages seeking answers to elementary questions. } Additionally, the use of "ASAP" } was a bad idea. } } JQuinn } } PS: I sent Jenny Wang's message to her Chairperson and UG Program Director.
°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°° Stefan Geimer MCDB Dept. Yale University P.O. Box 208103 New Haven, CT 06520-8103 U.S.A.
Sending Jenny Wang's message to her Chairperson and UG Program Director was, in my eyes, really unnecessary. Even if it was just a nice try from Jenny Wang to get her homework done, this is really overdone.
Regards,
Stefan
Jim Quinn wrote:
} Don - } } College students should not broadcast messages seeking answers to elementary questions. } Additionally, the use of "ASAP" } was a bad idea. } } JQuinn } } PS: I sent Jenny Wang's message to her Chairperson and UG Program Director.
°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°° Stefan Geimer MCDB Dept. Yale University P.O. Box 208103 New Haven, CT 06520-8103 U.S.A.
My feeling is that those of you who lack a positve and constructive response to a question that is posed, simply should not respond. I have seen plenty of queries by "experts" which could be answered rather simply by opening a book, but I certainly do not stoop to condescension.
Cavin Mooers, Research Assistant Vitreous State Laboratory The Catholic University of America Hannan Hall Washington, DC 20064 (202) 319-5346phone (202) 319-4469fax
I believe you may be looking Hanks' Balanced Salt Solution. We can get it for you if you are looking to buy it, or contact me direct if you just need some information on it.
Dr. Charles Duvic
--
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Shea Miller 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 } -----------------------------------------------------------------------.} } Greetings all; } can anyone provide the formulation for Hank's buffer? I have looked in all of my books (most, but not all of which, admittedly, lean towards plants) and ca } } thanks in advance } shea } } Dr. S. Shea Miller } Agriculture & AgriFood Canada } Eastern Cereal & Oilseed Research Centre } Rm. 2068, K.W. Neatby Bldg. } Central Experimental Farm } Ottawa, Ontario, } Canada K1A 0C6 } Phone: (613)759-1760 } Fax: (613)759-1701 } E-mail: millers-at-em.agr.ca
The killer in vibration impact on sensitive equipment is resonance. We (Schnabel Engineering) have worked on many projects with varying equipment vibration problems. The common denominator in all of them is that resonances in the source, the vibration path (whether geological or structural) and the receiver (the equipment and its mountings) combine to produce an impact that must be ascertained. It is certainly desirable for the impact to be determined in advance, because the range of mitigation techniques is broader then. However, various retrofits are available. The key is to use the appropriate retrofit.
Vibration, travelling in waves, is different than heat, and a solution of just packing "stuff" around the site is generally inadequate; sometimes it works, but that is then just dumb luck. I have used TEM equipment (in grad school) and know that the column for a 100kV microscope has a substantially different construction and configuration, and therefore substantially different vibration response from a 1.2MV microscope. The taller tower of the 1.2MV instrument will most probably have lower resonant frequencies than the 100kV instrument. I am not sure if such information (mechanical resonance) is available for them, but it certainly can be measured.
Mitigation techniques range from modification of the source, to barriers (trenches or caissons around the facility), to floating floors, to dynamic or passive absorbers. Each has its place, and the best (including cost!) solution may be a combination of the above. Again, the reduction of resonances is the key to successful vibration mitigation. I would be happy to discuss such solutions with those interested.
As a side note, I first became acquainted with this list about a year and a half ago, with respect to optical microscope standards. I am impressed with the quality and quantity of contributions to the list.
Regards,
Doug Anderson
Douglas A. Anderson, PhD Senior Consultant Schnabel Engineering Associates (http://www.schnabel-eng.com) 510 East Gay Street West Chester, PA 19380 Phone: 610 696-6066, Fax: 610 696-7771
} -----Original Message----- } From: Lesley S. Bechtold [SMTP:lsb-at-jax.org] } Sent: Wednesday, January 02, 2002 1:31 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Fwd: vibration isolation standards } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } } } Happy New Year to everyone! } } } } We are setting up a totally new EM/LM lab - being built from the ground } } up. We have told our engineers that we need to have a vibration-free } } environment for optimum equipment operation. They would like to know } } exactly what vibration is tolerable and what isn't. Are there any } } standards or measurements out there that detail what limits can be } } tolerated and what can't? } } } } Thank you! } } } } Lesley } } } } Lesley S. Bechtold } } Supervisor, Biological Imaging } } The Jackson Laboratory } } 600 Main St. } } Bar Harbor, ME 04609 } } 207-288-6191 } } Lesley S. Bechtold } Supervisor, Biological Imaging } The Jackson Laboratory } 600 Main St. } Bar Harbor, ME 04609 } 207-288-6191 } } } } This e-mail including attached files is confidential. Its transmission is solely as an accommodation for the benefit of the recipient. The recipient bears the responsibility for checking its accuracy against corresponding originally signed documents provided by Schnabel Engineering Associates, Inc. If you received this e-mail in error, its use is prohibited. Please destroy it and immediately notify postmaster-at-schnabel-eng.com
I have two sets of data to interpreted: 1) X-ray rocking curve data, to analyze the defect (damage) of the wafer surface. 2) XPS data, to trace how the wafer surface composition will change with the increasing of sputtering time.
I need to have some fundamental idea of these two theories to interpreted data. Do you have any recommendations of the textbooks, or other papers I should read?
Any kind of help will be appreciated!
Xianglin Li
Center for Advanced Material Department of Chemical Engineering University of Massachusetts, Lowell Xianglin_Li-at-student.uml.edu Tel: 978-934-3411
Looks like there are different opinions out there. Before this turns into a flame war, I would personally like to know what the general opinion is. I therefore propose a poll. Just send me an email with nothing but one of the following numbers in the subject line, and I will tally those votes and provide a summary in a week (if I get enough emails):
1) Students should work through their assignments alone. Let's not support laziness at all. 2) We should help them help themselves by pointing them to general microscopy books. 3) We should take their questions and point them to specific microscopy books dealing with their problems 4) We should encourage them to contact us offline so we can test their sincerity and then either help them or not. This should be posted on the server to avoid redundancy. 5) We should help them with their questions by trying to answer them in a general way 6) We should go all out and answer their questions as best as we can.
I hope the statistics will not be skewed by a zillion students who send me "6" as an answer ;-)
Any suggestions are welcome! I also promise not to misuse the email addresses!
} } } please send the email to mb-at-soft-imaging.com { { { { { {
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Oakley, Jeff [mailto:oakleyj-at-rayovac.com] Sent: Friday, February 08, 2002 11:46 AM To: Microscopy-at-sparc5.microscopy.com
Tom,
You are correct, I am not an educator. I also agree with you whole heartedly that when someone reads a text in search of information, they learn a great deal more than they had planned on... It happens to me every time I pick up a book in search of an answer to a question. I'm sure this probably was the intent of the instructor.
The point I should have made in my previous post is that instead of shutting someone down and "scolding" them (which is exactly what some of the listers did) for what appeared to be a Cliff's Notes research method, the person should have been guided to useful web pages or texts that would have made them find the answers for themselves (which other posters did - kudos to those).
It is possible to be helpful while at the same time not giving someone a free ride.
I don't think my batteries are dead, Tom, I just think we are operating at different voltages.
Jeff
-----Original Message----- } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] Sent: Friday, February 08, 2002 9:26 AM To: Oakley, Jeff Cc: Microscopy-at-msa.microscopy.com
At 10:28 PM 02/07/02 -0500, Beauregard wrote: } Polaroid did make a few of them. I saw a metal one, photocopied it } and measured it. I have the insert spec's but not at home here. I } could scan it and post it on my web page.
Hi Don,
I have scanned in the photocopy of the special insert for the 4X5 holder. I made a mistake about it being recessed. The SS45 comes with one (2¼x2¼?) holder that is recessed and that was what I recalled at home as being recessed. After looking at the photocopy of an official insert, I realized my mistake.
The holder is nothing but a totally flat piece of steel with 6 pins sticking up, a rectangular hole in it to allow transmitted light to shine through the negative, and along the one outside edge of the insert are two slots that fit into the two pins on the 4x5 holder.
I will post two JPG images of the insert(s) at:
http://www.westol.com/~beaurega/ss45.htm
I included a scanned image of a steel insert / holder that came standard with the scanner. Notice the two holders have identical slot alignment in my one image. Adjust the DPI of the image to get your laserjet printed insert image to line up with the two pins in the 4x5 holder. Then use this accurate template to make or have made the insert.
One could use a double layer of old film to make the equivalents of the pin posts to keep the film in register. The whole thing is painted black. Use a Dremel tool cut off wheel to make the slots.
I agree, Stefan. Not only overdone, but ugly. Since when did we become proctors of other people's courses? I was satisfied with Jenny's answer. If she had ever considered a career in electron microscopy, I'll bet she's reconsidering now.
Randy Tindall EM Core University of Missouri
-----Original Message----- } From: Stefan Geimer To: MSA Listserver Sent: 2/8/2002 8:39 AM
Sending Jenny Wang's message to her Chairperson and UG Program Director was, in my eyes, really unnecessary. Even if it was just a nice try from Jenny Wang to get her homework done, this is really overdone.
Regards,
Stefan
Jim Quinn wrote:
} Don - } } College students should not broadcast messages seeking answers to elementary questions. } Additionally, the use of "ASAP" } was a bad idea. } } JQuinn } } PS: I sent Jenny Wang's message to her Chairperson and UG Program Director.
°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°° Stefan Geimer MCDB Dept. Yale University P.O. Box 208103 New Haven, CT 06520-8103 U.S.A.
VLSI Standards is in the midst of releasing (the product is in beta at this juncture) a NIST Traceable, 100 nm pitch standard for use with SEMs. The accuracy is equal to or less than 1 nm in most cases. Besides 100 nm, it will also be certified for 4 other pitch values. It will come in various wafer / die form factors to be able to accommodate CD-SEMs utilizing automated handlers, as found in the Semiconductor and related industries.
Please contact me directly offline and I'd be glad to provide you information on this exciting new product.
Regards -
Marc Helvey Strategic Accounts Manager VLSI Standards, Inc. 3087 North First Street San Jose, CA 95134-2006 Phone: (408) 428-1800, ext. 108 E-Mail: marc.helvey-at-vlsistd.com {mailto:marc.helvey-at-vlsistd.com} Internet: http://www.vlsistandards.com
-----Original Message----- } From: Windland, Mark J (MN14) [mailto:Mark.Windland-at-honeywell.com] Sent: Friday, February 08, 2002 7:51 AM To: Microscopy-at-sparc5.microscopy.com
We need to purchase a new calibration standard for our SEM that gets us down to the next level. We need to have accuracy down to 0.010 microns. We have been using the Geller MRS-3 and now are considering purchasing the MRS-4 traceable standard. I believe this will give us what we need but I was wondering if there are other standards out there that are better, the same, worse? I need to hear from the calibration specialists out there. Thanks,
Mark Windland Honeywell Minneapolis, Minnesota 763-954-2845
I don't think it is a simple answer to your questions. If the request is like the subject of these discussions, then I would say number three. If the student has researched the basic information (showing an earnest effort at learning), and asks for help with understanding what they have found, or carrying it further, then I see no problem with the experts discussing and feeding information to the student. Giving them "food for thought" should not be a problem.
Darrell
Mike Bode {mb-at-Soft-Imaging.com} on 02/08/2002 06:19:42 PM
To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} cc:
Hello Listers:
Looks like there are different opinions out there. Before this turns into a flame war, I would personally like to know what the general opinion is. I therefore propose a poll. Just send me an email with nothing but one of the following numbers in the subject line, and I will tally those votes and provide a summary in a week (if I get enough emails):
1) Students should work through their assignments alone. Let's not support laziness at all. 2) We should help them help themselves by pointing them to general microscopy books. 3) We should take their questions and point them to specific microscopy books dealing with their problems 4) We should encourage them to contact us offline so we can test their sincerity and then either help them or not. This should be posted on the server to avoid redundancy. 5) We should help them with their questions by trying to answer them in a general way 6) We should go all out and answer their questions as best as we can.
I hope the statistics will not be skewed by a zillion students who send me "6" as an answer ;-)
Any suggestions are welcome! I also promise not to misuse the email addresses!
} } } please send the email to mb-at-soft-imaging.com { { { { { {
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Oakley, Jeff [mailto:oakleyj-at-rayovac.com] Sent: Friday, February 08, 2002 11:46 AM To: Microscopy-at-sparc5.microscopy.com
Tom,
You are correct, I am not an educator. I also agree with you whole heartedly that when someone reads a text in search of information, they learn a great deal more than they had planned on... It happens to me every time I pick up a book in search of an answer to a question. I'm sure this probably was the intent of the instructor.
The point I should have made in my previous post is that instead of shutting someone down and "scolding" them (which is exactly what some of the listers did) for what appeared to be a Cliff's Notes research method, the person should have been guided to useful web pages or texts that would have made them find the answers for themselves (which other posters did - kudos to those).
It is possible to be helpful while at the same time not giving someone a free ride.
I don't think my batteries are dead, Tom, I just think we are operating at different voltages.
Jeff
-----Original Message----- } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] Sent: Friday, February 08, 2002 9:26 AM To: Oakley, Jeff Cc: Microscopy-at-msa.microscopy.com
Dear all
I've been reading the discussions on this topic with interest, as a lecturer who sets similar EM assignment questions for undergraduate students.
Last year, a couple of my students posted their assignment questions to the listserver. After an initial feeling of annoyance that the students were being lazy, and not seeking out information for themselves, I eventually decided that this probably wasn't such a bad thing - as Mike said in his email, we are supposed to be teaching students to identify and utilize different sources of information!
In my case, I hadn't directly told the class about the listserver, so my students had either sought it out by themselves, or had actually read through a list of suggested reference texts and websites which had been given to them early in the semester. Either way, this was something to be encouraged!
I certainly don't think the listserver is the place where we should provide full and detailed answers to students' assignment questions - and particularly not when demanded "ASAP", and without evidence of the student having done any of their own homework on the topic!
However, I also share the views expressed in some earlier messages that we are a useful "resource" for students. For those who have the time and inclination to respond to students' requests, I support the approach of offering some basic information (brief, easy to understand) as a starting point, and then suggesting that the student refer to texts, papers or other sources. Indeed, this is exactly what happened with my students, and both presented assignments with information gleaned from a wide variety of sources. Many thanks to those of you who responded in this way.
Cheers Deb ***************************************************** Dr Deborah Stenzel Lecturer (Microbiology) School of Life Sciences and Applications Specialist (Biological) Analytical Electron Microscopy Facility Queensland University of Technology GPO Box 2434 Brisbane 4001 Australia
We have a perfectly good LKB Ultramicrotome ("Ultratome") Model #2088 (circa. 1976) for which we have no need. We have several others and we DO need the space! It also has most of the parts for a cryokit.
If anyone is interested: its yours for FREE! Just come and pick it up or arrange for shipment.
Please feel free to call or leave a message at any time. -- Peter Ingram Sr. Physicist, RTI Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
In much less time than it took to find addis for Jenny Wang's chairperson and program director he could have referred Jenny to Bozzola/Russell or any other EM text for the answers. This string has become more about those who want to educate and help vs. those who would judge and punish.
Still listening and learning. Pete Polsgrove
} ===== Original Message From Stefan Geimer {stefan.geimer-at-yale.edu} ===== } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Pete Polsgrove NAU Flagstaff, AZ. pjp6-at-dana.ucc.nau.edu micro2001p-at-netscape.net
I have been stunned by how many would deny answers to a student. I have to confess that I did not analyse the enquirer's motives, and offered some simple answers to her questions
Is the objection that the enquiry came via the internet, so was directed to all on the list? Would your reaction have been the same if the student made a more personal, targeted approach a) Called at your lab/office to ask questions b) Wrote asking questions c) Phoned/Faxed you asking questions
I think we have to acknowledge that the pre-internet world of the printed page and the post-internet world are totally different. Students are under immense pressure, not least under the burgeoning weight of the paper literature, and simply cannot afford the time to plough through roomfuls of books, however good this would be for their souls. They need entry points to a problem, and they should be congratulated for using all of the facilities currently at their disposal to get there. If that changes the way educators have to go about assessment of project work so be it. That is our professional problem, not the student's problem. This list has a significant educational role at all levels within research and tertiary education, and I would be saddened if barriers are erected against enquiries from students.
Dr. Chris Jeffree University of Edinburgh Biological Sciences EM Facility
As for "bad taste" from the MSA response, I taste no such displeasure.
I must say the questions asked were vague and rather general. Have I missed something? In all this communication, have we heard from the course professor explaining, defending, or otherwise making a statement as to what he or she was requiring of the student?! Why is not the professor explaining this material to the student? The professor should be the FIRST resource, or at least provide the student with a rudimentary understanding of the topic and perhaps assigning readings from handouts or materials on reserve in the library.
I teach a dedicated biomicroscopy course (optical light microscopy, TEM amd SEM) to upper division biology students, and have for 9 years. I would never just suggest students be turned out to fend on their own. I provide a a number of reserve textbooks and a huge number of handouts. I ALWAYS suggest if students are having a difficult time finding answers to questions I have proposed, they come to me FIRST!! In this manner, I have control over the ratio of student ability and information available.
I believe the hallmark of an educator is to entice and DIRECT the student in a manner of investigation, not to just through out a bunch of questions, allowing the student to randomly be come up with the answers, some of which depending of the source may be incorrect.
I must admit, when I saw the original email, my thoughts were divided into two direction: 1) here is a student who is looking for quick answers to some vague, rather general questions; and 2) here is a professor who is too busy with something else and has not put forth the foundation from which the student could asked specific questions about the general topics, e.g., "I have read this about the subject and do not understand. Is there someone on the listserver who can explain it differently from how I interpret the subject?"
Again, to some extent this is the responsibility of the professor.
Enough said...
Ken -- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203
} ===== Original Message From Dr Deborah Stenzel {d.stenzel-at-qut.edu.au} ===== Hello Group, I agree with Deb's post. Following this thread, I got to thinking that it is maybe time WE (as educator/citizens) do a little reading. Times they are a changin'. Try to find this article: Get Ready for the Net Generation, by Mark L. Alch, taken from the February 2000 issue of Training & Development. I found it in the Human Resources 01/02 Annual Editions, ISBN 0-07-243342-6. The way the new generations are learning happens to be a little different than the way most of us did. We not only have to understand how they learn, but we will also have to understand what motivates them once we hire them. Randy
} Dear all } Last year, a couple of my students posted their assignment questions } to the listserver. After an initial feeling of annoyance that the } students were being lazy, and not seeking out information for } themselves, I eventually decided that this probably wasn't such a bad } thing - as Mike said in his email, we are supposed to be teaching } students to identify and utilize different sources of information! } } In my case, I hadn't directly told the class about the listserver, so } my students had either sought it out by themselves, or had actually } read through a list of suggested reference texts and websites which } had been given to them early in the semester. Either way, this was } something to be encouraged! } } I certainly don't think the listserver is the place where we should } provide full and detailed answers to students' assignment questions - } and particularly not when demanded "ASAP", and without evidence of } the student having done any of their own homework on the topic! } } However, I also share the views expressed in some earlier messages } that we are a useful "resource" for students. For those who have the } time and inclination to respond to students' requests, I support the } approach of offering some basic information (brief, easy to } understand) as a starting point, and then suggesting that the student } refer to texts, papers or other sources. Indeed, this is exactly } what happened with my students, and both presented assignments with } information gleaned from a wide variety of sources. Many thanks to } those of you who responded in this way. } } } Cheers } Deb } ***************************************************** } Dr Deborah Stenzel } Lecturer (Microbiology) } School of Life Sciences } and } Applications Specialist (Biological) } Analytical Electron Microscopy Facility } Queensland University of Technology } GPO Box 2434 } Brisbane 4001 } Australia } } Phone + 61 7 3864 5036 } Fax + 61 7 3864 5100 } email d.stenzel-at-qut.edu.au } } http://www.sci.qut.edu.au/aemf
I can not believe all the unnecessary and self-righteous email one student has generated by asking a few questions! For those of you out there who feel it was "wrong" for a student to ask these questions, don't answer them. But as for emailing her Dept. head, when did this list become some sort of regulated police like forum? If this is what it has degraded to, I what off! As I tell my children and student alike there are no stupid or bad questions. As professional (at least I thought we all were before this) I believe that it is not only out duty but our obligation to help other learn. I don't mean doing there homework for them, but the response that this girl/women received embarrassed and ashamed me. I simple reply guiding her were to search for the answers could have saved everyone a lot of time and seem energy as well. As for people in general asking questions that the answers can be found in books, again if you don't want to answer them, don't. As for me, I am not the smartest man in the world nor do not know every thing and naively thought that this is what forum/list servers like this one were for. Guess I was wrong
Richard Cole Research Scientist III Director of the Laser Microsurgery and Advanced Light Microscopy Core Unit Wadsworth Center P.O. Box 509 Albany N.Y. 12201-0509 518-474-7048 Phone 518-486-4901 Fax
I looked into the background of the student's course early on and sent a brief explanation to the List, twice, but it never arrived, as far as I can tell. I'll tack in on below. One of the points of this biomedical engineering course that has been overlooked is that it is a PBL course.
PBL is a new philosophy of teaching that is being used, in part, by various medical schools and othe programs. I don't know a whole lot about it, but I know it exists because the University of Hawaii was the first place to embrace PBL wholly, instead of partially as at other institutions. Basically, the medical students here have no formal courses, but are thrown directly out into the clinics from day one, and are encouraged to learn what they need to know to do their "jobs" fairly independently. I'm sure this is an oversimplification, but not by a lot. There was a hue and cry from many of the instructors - how do you learn gross anatomy without a gross anatomy class and a cadaver? But I guess they worked it out. When I ask the current crop of students if they like it, they say they do. There's more of an emphasis on learning by doing and asking than just sitting in classrooms all day and with books all night.
However, here's the point - they are encouraged to go out and learn how to find answers in the world, using all resources from the library to asking experts to the Internet. And they are told not to just ask the teacher".
Do I agree with this method? Mostly no, probably because I didn't learn that way, and I'm old enough to be kinda set in my ways. . Is it working? Apparently yes. Now that I'm not an official student, is that the way I learn *now*? Yes, it is!
This is not an endorsement of the PBL system (y'all need to do some research on it to understand how the philosophy, as do I), but merely an explanation of why the students are asking the questions. And then ignore or guide them, whatever you wish.
Aloha, Tina
Message that did not reach the List:
I have received several emails recently about TEMs, as have several of you as well as this List. At first I dismissed tham as being at about the same level as Mrs. Jones' 6th grade science class who each individually emailed me to ask "How does an electron microscope work?" However, I looked into this and found out that this recent spate are from a Biomedical Engineering class at Georgia Tech. There are 60 students, split up into teams, involved in a Problem Based Learning curriculum, which encourages using all resources available, from the library to interviewing experts. Their project is an interesting one - although I deleted the original questions, I think it involves designing an original, viable improvement for the electron microscope, especially in areas that would allow them to create a 4D database of living cells and their cellular functions.
I received a couple of messages from students who had apparently done their library research and were able to ask thoughtful, reasoned questions. I feel that the ones who simply regurgitate the instructors' list of questions *do* need to do more background research and then formulate specific questions and direct them to the particular experts in the field. But they have only a couple of weeks on this assignment, so I guess I understand their "spamming" the List to find those experts!
I thought giving you all the background on the project would help you decide how to respond to the messages. It is an interesting mental exercise to think about how to build such an electron microscope. Perhaps they will!
**************************************************************************** * 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* ****************************************************************************
FYI: ctfExplorer has been updated and it is still free for all.
New features/improvements/fixes:
- Corrected an error in the formula used for Focal Spread calculation which caused too strong damping by temporal envelope at high frequencies. Thanks to Michael O'Keefe, Peter Tiemeijer and Uwe Lucken for pinpointing this error.
- Added a posibility to change values for high voltage and objective lens current instabilities (along with chromatic aberration and energy spread they affect the value of focal spread)
- Added a possibility of editing/saving/restoring of the microscope list
To the best of my knowledge, the software does not have any nasty bugs. It is tested under Windows 95/98/NT4/2000.
Please give it a try. Please DO direct your suggestions and comments to sidorov-at-yahoo.com I hope you'll find the software useful.
Here is the link: http://clik.to/ctfexplorer (always use this link. it will direct you to the right location). Direct link: http://www.maxsidorov.com/ctfexplorer
Enjoy, __________________________________ Max Sidorov, Ph.D. max.sidorov-at-amd.com
----------Additional Info---------- ctfExplorer is a highly interactive program which calculates/displays the contrast transfer function of TEMs. I know that there are similar programs floating around but ctfexplorer does not only 1d but also 2d calculations/display with 2-fold and 3-fold astigmatism imposed. There are other unique features to it. All parameters (defocus, voltage, Cs, etc) can be changed interactively.
Features - Calculates 1-Dimensional CTF - Calculates 2-Dimensional CTF - Calculates Defocus Map - Calculates point-to-point resolution, Lichte defocus and info limit - Shows the effects of 2-Fold and 3-Fold astigmatism - Allows to change Defocus, 2-Fold and 3-Fold astigmatism in real time - Shows what happens to 1D CTF in different directions when there's astigmatism - Displays the damping envelopes - Allows to select a microscope from a list of microscopes - Allows to create a custom microscope - Allows to change HT, Cs, Cc, Energy Spread, Convergence for any microscope - 2 modes of operation: CTF Explorer (to see everything) and CTF Plotter - Compares 2 microscopes or 2 settings for 1 microscope - Saves 1D CTF, 2D CTF and "Defocus Map" as bitmaps or metafiles - Exports 1D plots to tab-delimited text format
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hazrat.hussain-at-iw.uni-halle) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, February 10, 2002 at 11:35:50 ---------------------------------------------------------------------------
Question: Dear Sir, I have stained a block copolymer isothermally crystallized samples with RuO4 vapors. The block copolymers constitute poly(ethylene oxide)(semicrystalline block) and poly(perfluorohexylethyl methacrylate) as second block. The structure is as PFMA-b-PEO-b-PFMA PEO block length is 227 EO units(20000 g/mol) and each PFMA block was 5-7 PFMA units. The TEM morphology that we got from this sample was showing a layered or lamellar structure.There are dark lines and bright lines. Now I dont know, which block has been stained with RuO4. The second question is again the staining assigning problem, I got TEM pictures from solutions of the same block copolymers, the samples were stained again with the same dye. we got some spherical micelles in the TEM picture, but we dont know, the staining blocks. I hope u will consider answer my questions. I will be grateful with my best regards Hussain
Gee, maybe this whole thread is just a Sociology experiment to gauge the response of a small group of specialists to a contentious issue! For my $0.02 ($0.033 Canadian), pick up a copy of "Silicon Snake Oil" by Cliff Stoll. Relevant reading, I think.
Cheers,
Jim
-- James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Hussain Hazrat wrote: ============================================ Question: Dear Sir, I have stained a block copolymer isothermally crystallized samples with RuO4 vapors. The block copolymers constitute poly(ethylene oxide)(semicrystalline block) and poly(perfluorohexylethyl methacrylate) as second block. The structure is as PFMA-b-PEO-b-PFMA PEO block length is 227 EO units(20000 g/mol) and each PFMA block was 5-7 PFMA units. The TEM morphology that we got from this sample was showing a layered or lamellar structure.There are dark lines and bright lines. Now I dont know, which block has been stained with RuO4. The second question is again the staining assigning problem, I got TEM pictures from solutions of the same block copolymers, the samples were stained again with the same dye. we got some spherical micelles in the TEM picture, but we dont know, the staining blocks. I hope u will consider answer my questions. I will be grateful with my best regards Hussain ======================================================== This kind of question is extremely difficult to "call" on the basis of first principles (or logic). You really need to see one or two "knowns", and then you can see which way the area percent of the dark vs. white changes.
If you are seeing some "spherical" features from solution precipitation, and it is just a guess, it might be that you are seeing segregation of homopolymer into a more traditional kind of morphology for these kinds of systems. At least when we have seen such features in other block copolymer systems that was our conclusion.
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 ######################## ============================================
Just wanted to give a heads up for anyone with older Kevex LN2 cooled EDS systems. On Friday, our Kevex LN2 level sensor just exploded after 15years of service. Normally we remove the sensor (unplugged from the system) and place it horizontal in a special holder while we fill the LN2 then we dry it off and put it back in place. Today the metal top blew off and hit me in the leg. No injuries except the loud bang may have shaved a year or so off my life. Luckly, we had a second sensor on hand for replacement.
If any one has a good explanation why the metal cover decided to tear itself away from the Styrofoam insulation after all these years we would like to hear from you.
TIA
Jon Ekman Florida State University Biological Science Imaging Resource 119 Bio Unit I, 4370 Tallahassee, FL 32306 tel: 850.644.6519 fax: 850.644.0481
Let me inform you the list of microscopy laboratories at the "Petr's Microscopy Resources" has been completely rebuilt. You can check it at the http://www.petr.isibrno.cz/microscopy/laboratories.php .
Furthermore, the form for a new link submission has been revised to a great extent. Therefore, the addition of a new link to your laboratory is very easy and safe now, and your submission will be very appreciated. You can find the submission form at the new location http://www.petr.isibrno.cz/microscopy/PMRform.php .
Regards,
Petr Schauer +---------------------------------------------------------------------+ | Dr. Petr Schauer tel.: (+420 5) 41514313 | | Head of Electron Microscopy Laboratory fax : (+420 5) 41514404 | | INSTITUTE OF SCIENTIFIC INSTRUMENTS (+420 5) 41514402 | | ACADEMY OF SCIENCES OF THE CZECH REPUBLIC e-mail: petr-at-isibrno.cz | | Kralovopolska 147, CZ-612 64 Brno petr-at-dp.fce.vutbr.cz | | Czech Republic www: http://www.petr.isibrno.cz/ | +---------------------------------------------------------------------+
OK so it is a given that consumer grade cameras have relativly poor low light performance. That said, are there cameras that have better than average lowlight performance? Are any of the consumer cameras capable of binning?
Mike
"Mardinly, John" wrote: } } } John; } You are correct about the small CCDs of consumer digital cameras } have sever performance deficiencies due to their small pixel size. The F707 } and other consumer digital cameras that use the same chip (2/3"-Nikon 5000 } and Olympus E20N) suffer from noise even in visible light photographs. The } Nikon D1X and the new Canon EOS 1D(advertised 11.5 µm pixels) are } significant improvements, but cost $6000 just for the camera body! One } organization addressing this problem is } http://www.fredmiranda.com/Action_profilesPage/index.html where they discuss } and sell Photoshop plug-ins for noise reduction. Another digital camera site } I really like is: } http://www.imaging-resource.com/ }
--
________________________________________________________ / Michael J. Herron, U of MN, Dept. of Pediatrics/BMT / / herro001-at-umn.edu / / 612-626-4321 Mpls MN 55455 / /_______________________________________________________/
I see we have the equivalent of a prison snitch in our midst ..or more appropriately in this case ..a school yard tattle-tale Jim Quinn wrote:
} Don - } } College students should not broadcast messages seeking answers to elementary questions. } Additionally, the use of "ASAP" } was a bad idea. } } JQuinn } } PS: I sent Jenny Wang's message to her Chairperson and UG Program Director.
Paul D. Nolan Electron Optics
Alcan International Limited Kingston Research and Development Centre P.O.Box 8400, 945 Princess Street Kingston, Ontario K7L 5L9
yes, "Education is a bueaucracy, learning is a biological activity! The instructor serves as a resource with the ability to interact with and to direct the inquiry of the student, but learning happens only at the pleasure of the student. Sterling Stoudenmire, 1982.
At 10:27 AM 2/9/02 -0600, rnessler wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have been getting emails with responses, but right now the numbers are probably too low to be statistically significant. I'll wait until the end of the week.
Also, please note: I had requested the emails to be sent directly to me, because I did not want to overload the listserver. Again, please send the responses to mb-at-soft-imaging.com.
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
The only thing I could think that could cause this would be ice trapping LN2 in the sensor tube that then warmed up causing N2 gas pressure high enough to pop the top. The design of our sensors have the BNC connection on the top of the cap. Did the wires come out with the metal top?
} } Hi all, } } Just wanted to give a heads up for anyone with older Kevex LN2 cooled } EDS systems. On Friday, our Kevex LN2 level sensor just exploded after } 15years of service. Normally we remove the sensor (unplugged from the } system) and place it horizontal in a special holder while we fill the } LN2 then we dry it off and put it back in place. Today the metal top } blew off and hit me in the leg. No injuries except the loud bang may } have shaved a year or so off my life. Luckly, we had a second sensor } on hand for replacement. } } If any one has a good explanation why the metal cover decided to } tear itself away from the Styrofoam insulation after all these years } we would like to hear from } you. } } TIA } } Jon Ekman } Florida State University } Biological Science Imaging Resource } 119 Bio Unit I, 4370 } Tallahassee, FL 32306 } tel: 850.644.6519 } fax: 850.644.0481
-- David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
} ------------------------------------------------------------------------ } 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 see we have the equivalent of a prison snitch in our midst ..or more } appropriately in this case ..a school yard tattle-tale } Jim Quinn wrote: } } } Don - } } } } College students should not broadcast messages seeking answers to } elementary questions. } } Additionally, the use of "ASAP" } } was a bad idea. } } } } JQuinn } } } } PS: I sent Jenny Wang's message to her Chairperson and UG Program } Director. } } Paul D. Nolan } Electron Optics } } Alcan International Limited } Kingston Research and Development Centre } P.O.Box 8400, 945 Princess Street } Kingston, Ontario K7L 5L9 } } Tel: (613) 541-2066 } Fax: (613) 541-2134 } paul.nolan-at-alcan.com
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 **********************************************
Hi Listers, I have a client who is writing a grant and has "re-discovered" some old techniques that could be very useful in her research. The problem is, I'm having trouble finding a source or sources for the reagents. Any ideas on where we could get the following? Thoria Sol ( aqueous colloidal thorium, once sold by Polysciences. It does not appear in their on-line catalog, nor in any of the other catalogs I've checked) to be used for pinocytotic uptake to label lysosomes. We could use ferritin, but that's so messy (in my hands, anyway). the full protocol for Gomori's method of acid phosphatase labelling. I have the citation on order from Inter-Library loan (Arch. Pathol.1941!!!) but don't know when it will come it. Are the reagents still available? Has anyone out there done either of these techniques? Any suggestions for alternates (preferably not immuno)? Thanks a million, Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Couldn't find a paper copy in our library so I downloaded a synopsis from the web, along with the reviews. Very, very funny, but not true. His dinosaurian heritage is showing, and paper is definitely dead. Silicon is the world's commonest material, not cellulose. Scientific publishing has ripped us all off. We have ownership of the content, we did all the work, and they charge us so much for the journals our libraries cannot afford the subscriptions. Nor can universities afford the upkeep of the libraries. There used to be departmental libraries here in every department. Not any more. There was a time when Universities could afford the upkeep of their physical establishments. Now they're selling of the paintings to keep up with the maintenance. The revolution is coming, and when it does paper journals will be first against the wall. Twenty years from now, maybe sooner, scientists will publish online, and the last 50 years of publishing will be accessible online from anywhere in the world, and many tertiary education courses will be distributed, campusless, attended by students wherever they happen to live in the world. And the educators? They'll be made by Intel ......
jmtc Chris
} Gee, maybe this whole thread is just a Sociology experiment to gauge } the response of a small group of specialists to a contentious issue! For } my $0.02 ($0.033 Canadian), pick up a copy of "Silicon Snake Oil" by } Cliff Stoll. Relevant reading, I think. } } Cheers, } } Jim } } -- } James M. Ehrman } Digital Microscopy Facility } Mount Allison University } Sackville, NB E4L 1G7 } CANADA } } phone: 506-364-2519 } fax: 506-364-2505 } email: jehrman-at-mta.ca } www: http://www.mta.ca/~jehrman } } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Waddington Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JN, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
Mike, #3, keeping in mind the possible limited sources available to the student (high school students may not have *any* EM books available in their library---so they need to be pointed to alternate information sources). Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences Binghamton University Binghamton, NY 13902-6000
-----Original Message----- } From: Mike Bode [mailto:mb-at-soft-imaging.com] Sent: Friday, February 08, 2002 6:20 PM To: 'Microscopy-at-MSA.Microscopy.Com'
Hello Listers:
Looks like there are different opinions out there. Before this turns into a flame war, I would personally like to know what the general opinion is. I therefore propose a poll. Just send me an email with nothing but one of the following numbers in the subject line, and I will tally those votes and provide a summary in a week (if I get enough emails):
1) Students should work through their assignments alone. Let's not support laziness at all. 2) We should help them help themselves by pointing them to general microscopy books. 3) We should take their questions and point them to specific microscopy books dealing with their problems 4) We should encourage them to contact us offline so we can test their sincerity and then either help them or not. This should be posted on the server to avoid redundancy. 5) We should help them with their questions by trying to answer them in a general way 6) We should go all out and answer their questions as best as we can.
I hope the statistics will not be skewed by a zillion students who send me "6" as an answer ;-)
Any suggestions are welcome! I also promise not to misuse the email addresses!
} } } please send the email to mb-at-soft-imaging.com { { { { { {
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Oakley, Jeff [mailto:oakleyj-at-rayovac.com] Sent: Friday, February 08, 2002 11:46 AM To: Microscopy-at-sparc5.microscopy.com
Tom,
You are correct, I am not an educator. I also agree with you whole heartedly that when someone reads a text in search of information, they learn a great deal more than they had planned on... It happens to me every time I pick up a book in search of an answer to a question. I'm sure this probably was the intent of the instructor.
The point I should have made in my previous post is that instead of shutting someone down and "scolding" them (which is exactly what some of the listers did) for what appeared to be a Cliff's Notes research method, the person should have been guided to useful web pages or texts that would have made them find the answers for themselves (which other posters did - kudos to those).
It is possible to be helpful while at the same time not giving someone a free ride.
I don't think my batteries are dead, Tom, I just think we are operating at different voltages.
Jeff
-----Original Message----- } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] Sent: Friday, February 08, 2002 9:26 AM To: Oakley, Jeff Cc: Microscopy-at-msa.microscopy.com
I vaguely recall the cap coming off of our Kevex cap years ago, but without near so much excitement. I think it just worked loose. A check of the wires and a little bit of epoxy and the cap was on again and has been fine since.
I can't remember if the dipstick was glued tightly into the Styrofoam plug. If it was, I could see pressure building up under the cap. I don't think the metal on ours was not glued all the way around.
Warren
At 11:25 AM 2/11/02 -0500, you wrote:
} The only thing I could think that could cause this would be ice trapping } LN2 in the sensor tube that then warmed up causing N2 gas pressure high } enough to pop the top. The design of our sensors have the BNC connection } on the top of the cap. Did the wires come out with the metal top? } } } } } Hi all, } } } } Just wanted to give a heads up for anyone with older Kevex LN2 cooled } } EDS systems. On Friday, our Kevex LN2 level sensor just exploded after } } 15years of service. Normally we remove the sensor (unplugged from the } } system) and place it horizontal in a special holder while we fill the } } LN2 then we dry it off and put it back in place. Today the metal top } } blew off and hit me in the leg. No injuries except the loud bang may } } have shaved a year or so off my life. Luckly, we had a second sensor } } on hand for replacement. } } } } If any one has a good explanation why the metal cover decided to tear } } itself away from the Styrofoam insulation after all these years we would } } like to hear from } } you. } } } } TIA } } } } Jon Ekman } } Florida State University } } Biological Science Imaging Resource } } 119 Bio Unit I, 4370 } } Tallahassee, FL 32306 } } tel: 850.644.6519 } } fax: 850.644.0481
To answer your question you need to know, among other things, the specificity of interaction between RuO4 and the comonomers that comprise your block copolymer. I suggest that you consult the literature for reactivity of RuO4 with your materials. I usually start with Sawyer and Grubbs book, Polymer Microscopy. I know that the first edition has information that should help you.
Good luck,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Garber, Charles A." To: MICROSCOPY BB {cgarber-at-2spi.c {Microscopy-at-sparc5.microscopy.com} om} cc: Subject: staining of block copolymers
02/11/02 07:40 AM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Hussain Hazrat wrote: ============================================ Question: Dear Sir, I have stained a block copolymer isothermally crystallized samples with RuO4 vapors. The block copolymers constitute poly(ethylene oxide)(semicrystalline block) and poly(perfluorohexylethyl methacrylate) as second block. The structure is as PFMA-b-PEO-b-PFMA PEO block length is 227 EO units(20000 g/mol) and each PFMA block was 5-7 PFMA units. The TEM morphology that we got from this sample was showing a layered or lamellar structure.There are dark lines and bright lines. Now I dont know, which block has been stained with RuO4. The second question is again the staining assigning problem, I got TEM pictures from solutions of the same block copolymers, the samples were stained again with the same dye. we got some spherical micelles in the TEM picture, but we dont know, the staining blocks. I hope u will consider answer my questions. I will be grateful with my best regards Hussain ======================================================== This kind of question is extremely difficult to "call" on the basis of first principles (or logic). You really need to see one or two "knowns", and then you can see which way the area percent of the dark vs. white changes.
If you are seeing some "spherical" features from solution precipitation, and it is just a guess, it might be that you are seeing segregation of homopolymer into a more traditional kind of morphology for these kinds of systems. At least when we have seen such features in other block copolymer systems that was our conclusion.
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 ######################## ============================================
The Department of Geosciences at the University of Massachusetts invites applications for a Post-Doctoral Position in Geology. This two-year position is specifically aimed at the rapidly emerging techniques of electron microprobe analysis in geochronologic applications. UMass is currently developing an optimized electron microprobe with Cameca, France that is specifically designed for the exploration of techniques for age mapping and dating of minerals (e.g. monazite, zircon) and trace element analysis. This project includes optimization on virtually all fronts, hardware, software, and technique development. One future direction will involve synthesis and analysis of standards for calibration and background measurement studies. The successful applicant will collaborate with UMass Geosciences faculty (and associates) and with Cameca, and will be directly involved with improvements and modifications to software, continued evaluation of analytical techniques, synthesis and characterization of standard materials, and application of the new techniques to geologic problems. Applicants must have completed a Ph.D. in Geology, materials science, or other physical science, with preference given to those with significant experience in electron microprobe analysis, x-ray spectrometry, materials microanalysis, and/or scientific programming.
Please send a letter of application, resume, and two reference letters to Michael L. Williams, Dept. of Geosciences, University of Massachusetts, 611 North Pleasant Street, Amherst, MA 01003-9279. The University of Massachusetts is an Equal-Opportunity Affirmative -Action Employer; women and members of minority groups are encouraged to apply.
Review of applicants will begin March 15th; the position will remain open until a successful candidate is identified.
**************** Michael J. Jercinovic Assistant Professor Department of Geosciences University of Massachusetts 611 North Pleasant Street Amherst, MA 01003-9297 E-Mail: mjj-at-geo.umass.edu Phone: (413) 545-2431 http://www.geo.umass.edu/faculty/jercinovic.html
Electron Microprobe Laboratory http://www.geo.umass.edu/probe/probe.html
For a look at what's coming down the path, see today's (Monday, February 11th) New York Times article on Foveon or that company's website. (I have no financial interest in the company and was previously unaware of them.)
John Twilley Conservation Scientist
Michael Herron 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 } -----------------------------------------------------------------------. } } } All, } } OK so it is a given that consumer grade cameras have relativly poor low } light performance. That said, are there cameras that have better than } average lowlight performance? Are any of the consumer cameras capable } of binning? } } Mike } } } "Mardinly, John" wrote: } } } } } John; } } You are correct about the small CCDs of consumer digital cameras } } have sever performance deficiencies due to their small pixel size. The F707 } } and other consumer digital cameras that use the same chip (2/3"-Nikon 5000 } } and Olympus E20N) suffer from noise even in visible light photographs. The } } Nikon D1X and the new Canon EOS 1D(advertised 11.5 µm pixels) are } } significant improvements, but cost $6000 just for the camera body! One } } organization addressing this problem is } } http://www.fredmiranda.com/Action_profilesPage/index.html where they discuss } } and sell Photoshop plug-ins for noise reduction. Another digital camera site } } I really like is: } } http://www.imaging-resource.com/ } }
Hello, I am setting up a TEM/SEM lab and I need a vacuum coater and critical point freeze drier for the SEM. I cannot afford new equipment and I will consider any used but still functioning equipment, complete with manuals.
Thank you.
Greg Barclay
Dr.G.F. Barclay Plant Science Unit, Dept. of Life Sciences University of the West Indies St. Augustine, Trinidad and Tobago, West Indies
I'm trying to prepare tem samples from sapphire. There is a thin metal film on the sapphire substrate, as well. I don't have too much experience in this field so, I would greatly appreciate any suggestions about preparing tem samples from sapphire.
Previously, I have prepared couple of Si samples but, sapphire seems to be much harder and difficult to deal with.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cassel-at-biology.queensu.ca) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, February 11, 2002 at 15:05:08 ---------------------------------------------------------------------------
Email: cassel-at-biology.queensu.ca Name: Stephen Casselman
Organization: Queen's University
Education: Graduate College
Location: Kingston, Ontario, Canada
Question: I am involved in a project which is examining sperm in fish, we will be using a video camera and a microscope to film motile sperm. Much of this work will be done in remote locations. We are interested in buying a new microscope that would able to handle frequent transportation to these remote locations. Ideally the scope would have a padded case specifically for it to be transported in. We generally use a 40 X objective lense. Does such a durable scope exist?
} Hi Listers, } I have a client who is writing a grant and has "re-discovered" some } old techniques that could be very useful in her research. The } problem is, I'm having trouble finding a source or sources for the } reagents. Any ideas on where we could get the following? } Thoria Sol ( aqueous colloidal thorium, once sold by Polysciences. It } does not appear in their on-line catalog, nor in any of the other } catalogs I've checked) to be used for pinocytotic uptake to label } lysosomes. We could use ferritin, but that's so messy (in my hands, } anyway).
How about peroxidase followed by a DAB reaction? Shows pinocytosis well. The method should be in Hyatt's book? Most histology texts (Weiss for one) will have an EM of capillary endothelium with peroxidase showing the vesicles. It may be a Karnovsky technique.
} the full protocol for Gomori's method of acid phosphatase labelling. } I have the citation on order from Inter-Library loan (Arch. } Pathol.1941!!!) but don't know when it will come it.
Gomori had a book out about 1950 or so, I suspect your library will have it. Also, any edition of Lillie's "Histopathological Technique and Practical Histochemistry" should do. Also John Kiernan's book should have it. Maybe even Humason's book will have it.
} Are the reagents still available?
Fisher, Sigma, Aldrich.
} Has anyone out there done either } of these techniques?
Not since grad school.
} Any suggestions for alternates (preferably not } immuno)? } Thanks a million, } Lee } } -- } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175
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 was just sitting back (but intrigued) but now I must share that I had a very similar thought as your own.
Yeah, I imagined Jenny's actual assignment was to conduct a psychology experiment, the question refined beautifully to elicit a response. Maybe we should name this technique to gauge personality and opinion after her....The Jenneric Response Factor. It might even be good for extra credit on her report. anyway, it gave me quite a chuckle. Even if the original intent was not to extract a slice of humanity, it will perhaps be the most valuable life lesson. Fun stuff.
antiflame disclamer: I am not making light of anyone's serious and passionate responses, just a perspective.
I also find it interesting which questions generate the most responses on the listserver.
Regards, Ed
"James M. Ehrman" {jehrman-at-mta.ca} on 02/11/2002 04:48:27 AM
To: Microscopy-at-sparc5.microscopy.com cc:
Gee, maybe this whole thread is just a Sociology experiment to gauge the response of a small group of specialists to a contentious issue! For my $0.02 ($0.033 Canadian), pick up a copy of "Silicon Snake Oil" by Cliff Stoll. Relevant reading, I think.
Cheers,
Jim
-- James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
} Any ideas on where we could get the following? } Thoria Sol ( aqueous colloidal thorium, once sold by Polysciences. It } does not appear in their on-line catalog, nor in any of the other } catalogs I've checked) to be used for pinocytotic uptake to label } lysosomes.
Hello Leona,
If you have it available, take a look at pp 116-117 (Section 3.5.5c "Staining acidic carbohydrates with colloidal thorium dioxide") of PR Lewis & DP Knight (1992) "Cytochemical staining methods for electron microscopy", Volume 14 of the "Practical Methods in Electron Microscopy" series.
They state that colloidal thorium was also known as Thorotrast, and was formerly used for medical X-ray diagnosis but is now difficult to obtain (it proved carcinogenic in the patients). Although Lewis & Knight refer to Thorotrast as 'colloidal thorium', other sources indicate that it is colloidal thorium dioxide - go to: http://brighamrad.harvard.edu/Cases/bwh/hcache/161/full.html
Lewis & Knight offer a recipe for home-made colloidal thorium dioxide as an alternative (CARE: RADIOACTIVE), and I have used this exact method myself to stain acidic carbohydrates. It worked a treat. Perhaps it would work in your application too? To make the thorium dioxide I used a very old bottle of thorium nitrate from BDH - a quick search of the WWW suggests it is no longer in their catalogue. Perhaps safety and disposal concerns make it difficult to obtain today. The EM labs on your campus might have a bottle tucked away, if you still want to try it.
Back to the ferritin perhaps? ;-)
Regards
Stephen Edgar
Pathology Department Faculty of Medicine & Health Sciences University of Auckland Private Bag 92019 Auckland New Zealand
} Hi Listers, } I have a client who is writing a grant and has "re-discovered" some } old techniques that could be very useful in her research. The } problem is, I'm having trouble finding a source or sources for the } reagents. Any ideas on where we could get the following? } Thoria Sol ( aqueous colloidal thorium, once sold by Polysciences. It } does not appear in their on-line catalog, nor in any of the other } catalogs I've checked) to be used for pinocytotic uptake to label } lysosomes. We could use ferritin, but that's so messy (in my hands, } anyway).
This is thorium dioxide, I think. Rather toxic, carcinogenic and radioactive (alpha-emitter) too. It's in the Alfa Aeser catalogue (www.alfa.com) under thorium (IV) oxide.
} the full protocol for Gomori's method of acid phosphatase labelling. } I have the citation on order from Inter-Library loan (Arch. } Pathol.1941!!!) but don't know when it will come it. } Are the reagents still available? Has anyone out there done either } of these techniques? Any suggestions for alternates (preferably not } immuno)?
In "Plant Cell Biology: a Practical Approach" (1994), p. 62, is a full protocol for doing this - I dug this up once before for a student. The authors say there are several methods based on the Gomori reactions, and this is one of them, and that a variety of substrates can be used, giving different coloured products for LM. It looks very straightforward, I guess you'd just have to be careful of artefacts. Ingredients: acetate buffer (acetic acid + Na acetate), naphthol AS-MX phosphate, Fast Red TR, Tris-HCl buffer, dimethylformamide.
If you're after a TEM protocol, "Electron Microscopy of Plant Cells" (1991) gives details on pp. 125-131, recipe p. 161, in which case Pb is precipitated in the reaction - also derived from Gomori. Many cautions about artefacts. Ingredients: beta-glycerophosphate, acetate or Tris-maleate buffer, lead nitrate solution. Or, can use cerium chloride in acetate buffer preincubation, this buffer plus beta-glycerophosphate stain.
Don't think there would be a huge difference between plant and animal (incl human!) cells....
cheers,
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
It is CMOS, not CCD, as I read it. This leads to an entirely different venue.
gary g.
At 12:50 PM 2/11/2002, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I just want to point out two problems with either of these camera, and with other too, from the cheapest to the most expensive :
1- Someone point out the probleme with the JPG compression. Right, but in practice you are obliged to use the best resolution to be alowed to save in TIFF format. If you need some memory to take a lot of pictures (dynamic process), or if you don't need 1500x2000 pixel, you cannot have the tiff format. And why don't these "new", "modern" camera use formats like JEPEG 2000 or SPIFF which let the choice of compressing (lossy or lossless) or not ?
2- An other point, more discreet, is that the picture is adjusted to the "best" dynamic scale before saving. The brighter pixel will be put to "white", i.e. level 255 in 8 bit BW , and the darker to "black", i.e. level 0, or something so. It's probably more sophisticated than that. The important result is that you CANNOT make quantitative mesures on brightness between different pictures. This is never said in commercial or technical shits. You can choice between auto adjustement, normal (what does it mean ?), more or less contrast or brightness, but you cannot put that fonction off (see p 104 of the Coolpix 995 manual). Cheapest camera have no settings, and do simply that adustement. More expensive one let you choice "something" but don't say what they really do. We have our Coolpix only since two month, so I had no time to try if there is a way to bypass this problem. Has someone a experience about that ? We had soon the same problem with the Fuji FinePix S1Pro. But we made only short tests with it.
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
For those who may bee interested, I have such a receipt (which I sent to Peter). Please ask off-line, an I can send it.
If some one else have one, I am too interrested. Our receipt works, but has its deffects !
By the way, Edwards remarked about an other topic (EM quest. ...)
"I also find it interesting which questions generate the most responses on the listserver." ..and which questions generate less (or no) responses.
I find it's difficult to know when it's useful to give an answer, when it is better to give it off-line or on the list. Those who use the list since years have perheps an advice about that.
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
The "MATERIA NOVA a.s.b.l." research Center, located in Mons - Belgium
recruits
1 Ph.D. or civil engineer - chemist or physicist
to set up and to be in charge of its Department of electronic microscopy (SEM and TEM)
Initiated in 1995 in close collaboration with the University of Mons-Hainaut and the Polytechnic Faculty of Mons, Materia Nova got its own identity in 2000 and has opened up largely its activities to the industrial world. Materia Nova activities rely upon two main research fields: interfacial aspects of materials and polymeric materials.
The main objectives of the Department of electronic microscopy can be summarized as follows: + To reinforce the research Center capacity in materials analysis and characterization : ion and electron spectroscopy - local probe microscopy - ellipsometry - calorimetric methods - optical spectroscopy - chromatographic methods - ... + To contribute to the formation/education in the field of surface and interface chemistry.
Letter of application (preferably written in French) and C.V. have to be addressed to:
Monsieur Joseph LEMINEUR - General Manager - MATERIA NOVA a.s.b.l., Parc Initialis, Avenue Nicolas Copernic, B-7000 Mons, Belgium e-mail: joseph.lemineur-at-umh.ac.be Phone : ++32 (0)65 373800
To find more information concerning MATERIA NOVA" research center, please visit : http://www.materia-nova.com
----------------------------------------------------------- Rachel Gouttebaron laboratoire d'Analyses de Surfaces par Spectroscopie Ionique et Electronique (LASSIE) Materia Nova Parc Initialis Avenue Nicolas Copernic 7000, mons, belgium tel : +32 65 37 38 52 fax : +32 65 37 38 41 e-mail : rachel.gouttebaron-at-umh.ac.be
I have been examining immunogold labelled cell and virus surfaces in a Hitachi S4700 FEG SEM, and would be grateful for your help/ wisdom in interpreting what I see. In SE mode, labelled sites are decorated with rounded blobs about 45-50nm diameter. Clear 10nm gold particles are visible by YAG BSE imaging in the centre of each blob. Blobs/gold are absent in unlabelled controls. Presumably the blob represents the IgG shell surrounding the gold particle? What is the "official" diameter for this shell? (I am trying to estimate how much I have grown it by carbon coating). At least as many labelled locations as are labelled with single gold probes are labelled with binary or ternary probes. Mostly these are pairs or triplets of overlapping blobs, with gold particle centres separated by, typically, 22-24 nm. Occasionally two gold particles appear to be very close together (i.e. touching or separated by 1-3 nm) at the centre of what appears to be a single blob. I assume these latter represent pairs of gold particles that were cross-linked by protein at the time of manufacture of the conjugate. Would that be a fair conclusion? Intermediate spacings seem infrequent (I haven't done any stats on this). Does anyone have a handle on how close individual gold probes can approach each other before being sterically excluded? This would have a bearing on the quantitative relationship between label numbers and closely-spaced epitopes. Is the answer by any chance somewhere in the 22-24 nm region?
Many thanks in advance Chris
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Waddington Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JN, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
A Short Course With Emphasis on Recent Innovations in Tools and Methods
(Including tripod polishing, ion milling, and FIB techniques.)
Instructors: Ron Anderson, IBM (retired); Fred Stevie, NC State; Lucille Giannuzzi, UCF
At the University of Central Florida (prior to the FL AVS/FL Society for Microscopy Meeting) Orlando, FL
Friday, Saturday and Sunday, March 8,9,10, 2002
for registration information please contact: Lucille Giannuzzi, lag-at-mail.ucf.edu
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 882-1500 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
If you have a Mac, try the latest version (or at least higher than 4.0.9) of Graphic Converter. http://www.lemkesoft.com shareware, an excellent graphics file-converter program with basic image-manipulation tools. For PCs, there's Irfanview http://www.irfanview.com/english.htm freeware, but there's a more complete shareware version, I *think*. I don't know if Irfanview will convert Gatan to other formats, though.
Phil
} Can anybody help with file conversion? } I need to convert Gatan Format (.dm3) file from Digital Micrograph to RAW } format. } } Thanks, } } Edy Widjaja } Materials Science and Engineering } Northwestern University } reply to : e-widjaja-at-northwestern.edu } office : 847-491-7809 lab : 847-491-3281 } http://www.numis.nwu.edu/internet/Staff/edy -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
This year we will be hosting the 7th annual RMC Materials Microtomy Short Course in Tucson, Arizona from April 16-19, 2002.
For those of you residing in colder climes, Tucson has just the weather to chase away the onset of those winter-time blues.
Join us, and our internationally renowned course faculty, in sunny Tucson to participate in this unique event. This short course is designed specifically for researchers in the field of materials science who wish to gain exposure to advances in specimen preparation for electron microscopy.
Please email me at stacy-at-boeckeler.com to receive full details and a course brochure.
Warm Regards,
Stacy Darnell Administrative Assistant RMC Products Division Boeckeler Instruments, Inc. stacy-at-boeckeler.com 4650 S. Butterfield Drive Tucson, AZ 85714, USA Tele: 520-745-0001 Fax: 520-745-0004
Hi Listers Thanks to all of you who responded to my inquiry. As always, you all have risen to the challenge, but after further discussion, my client is going to try an approach using fluorescent labelling and FRAP on the confocal. Neither of us wanted to deal with thorium if we didn't have to. If she does need the acid phosphatase for the EM. we'll do an AB-DAB method.
Lee -- Lee Cohen-Gould Electron & Optical Microscopy Facilities Weill Medical College of Cornell U. (212)746-6146 Rms A-105, LC-207
For a fairly easy read regarding CMOS vs. CCD technology, please check the following PDF.
The article is a reprint from Photonics Spectra, January 2001. I'm sure a few things may have changed, but the general ideas have held for a while, and continue to do so.
*Kind Regards, *Dave Hall *Resolution Technology, Inc - (614) 921-0045
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Tuesday, February 12, 2002 12:27 AM To: John Twilley Cc: MSA listserver
It is CMOS, not CCD, as I read it. This leads to an entirely different venue.
gary g.
At 12:50 PM 2/11/2002, you wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can anyone put me in touch with a source for "clam shell" interlocking gold alloy planchets used in the Balzers High Pressure Freezing apparatus - HPM 010? My search for the Swiss Precision company (our previous source) yielded an address in Palo Alto CA (908 Industrial Drive) and a phone number that proved to be that of a private home. The number associated with the company that was given on a web site that accurately described the planchets as Craig Type and as a product of that company was 650-493-0440.
Thanks for any help you can give me in tracking these folks down.
Regards, Bill Sharp William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Waddington Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JN, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
} } "James M. Ehrman" {jehrman-at-mta.ca} on 02/11/2002 04:48:27 AM } ... pick up a copy of "Silicon Snake Oil" by } Cliff Stoll. Relevant reading, I think. }
I've met Cliff Stoll. He's a very interesting character, and an archetypal hacker (in the classic favorable sense of technically proficient and inventive, before the word was co-opted by the ignorant press to mean people who break into computer systems). That someone with his background should be so negative about on-line communities is unusual. His "Cuckoo's Egg" book on computer security makes a dry subject very entertaining.
For another perspective, look at "The Cathedral and the Bazaar" by Eric Raymond. While the nominal subject is the open-source and Linux paths for software development, he is a keen observer of the behavior of on-line groups. His key insight is that, rather like academia but much less structured, reputation is the coin of the realm online as well, which is why so many people will spend so much time in what seems like unproductive activity in traditional career terms. Posts pile up as "publications" of a sort. Peer review is immediate and severe, as we've seen. :)
The New England Society for Microscopy announces that the next meeting of the Society will be held at 6.00 p.m. on Tuesday March 12th. 2002 at the Lexington Laboratory of Raytheon Corporation. The theme of the program will be Scanned Probe Microscopies.
Full details of the meeting, including the program, and information about the New England Society may be found on the Society's Web pages, located at http://www.msa.microscopy.com/MSALAS/NESM/NESMHome.htm
All interested persons are welcome, and invited to attend.
We have a few application notes on preparing sapphire for TEM that you can download from our website. Go to www.southbaytech.com and navigate to "Applications Support". Then select "Application Notes". You are looking for Application Notes 34 and 55. Number 34 deals with Tripod Polishing and number 55 deals with MicroCleaving.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
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'm trying to prepare tem samples from sapphire. There is a thin metal film on the sapphire substrate, as well. I don't have too much experience in this field so, I would greatly appreciate any suggestions about preparing tem samples from sapphire.
Previously, I have prepared couple of Si samples but, sapphire seems to be much harder and difficult to deal with.
Thank you very much, Ayten C. Aktas.
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
I will be preparing both plane view and cross sectional samples. My main concern is to keep the metal film in its initial state throughout the sample preparation steps (as much as possible). The metal films are generally 200-300 Angstrom thick.
Can dimpler alter/damage the thin film?
Second problem is the hardness of the sapphire. Do I have to use diamond products to reduce the thickness of the sapphire substrates? The substrates are 0.5 mm to start with.
For grinding and polishing few different type of equipment are available (from hand polising to automatic polishing machines). Getting access to equipment is not the main concern.
Thanks Ayten C. Aktas
On Mon, 11 Feb 2002, Ayten Celik 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 } -----------------------------------------------------------------------. } } } } } Dear all, } } I'm trying to prepare tem samples from sapphire. } There is a thin metal film on the sapphire substrate, as well. } I don't have too much experience in this field so, I would greatly } appreciate any suggestions about preparing tem samples from sapphire. } } Previously, I have prepared couple of Si samples but, sapphire seems to be } much harder and difficult to deal with. } } } Thank you very much, } Ayten C. Aktas. } }
Hi all, Does anyone have experience using polypyrrole films in lieu of formvar/collodion in TEM applications? They sound like the best thing since sliced bread, as they are very thin and conductive. Too good to be true? Comments? Randy
----- Forwarded by Gerard D Gagne/LAKE/PPRD/ABBOTT on 02/12/02 05:28 PM -----
Jane A Fagerland To: Gerard D Gagne/LAKE/PPRD/ABBOTT-at-ABBOTT cc: 02/12/02 Subject: internships 05:25 PM
The Department of Microscopy and Microanalysis at Abbott Laboratories will sponsor two summer internships for students interested in experiencing microscopy in the healthcare industry. One internship will be in Biological Microscopy, and one will be in Materials/Surface Science.
The department houses state-of-the-art equipment including field emission and environmental scanning electron microscopes with energy dispersive x-ray spectroscopy, transmission electron microscopes, several types of fluorescence technologies (including conventional fluorescence, confocal, and flow cytometry), light microscopy (including polarized light and interference contrast), laser capture microdissection, and all ancillary preparatory equipment such as microtomes, cryostat, and evaporative coaters. Surface analytical capabilities include x-ray photoelectron spectrometry and atomic force microscopy. We have several image analysis systems, including MetaMorph and AnalySIS. The department consists of nine microscopists with expertise in cell biology, materials analysis, surface chemistry, ultrastructural pathology, in vitro toxicology, and all the associated technical skills.
For details about summer internships at Abbott and for application forms and instructions, please visit www.abbott.com and follow the instructions there. Please note: the deadline for applications is March 1!
IN ADDITION, please send a copy of your resume with cover letter either by snail mail or e-mail to:
Jane A. Fagerland, Ph.D. Abbott Laboratories D-R45M/AP31 200 Abbott Park Rd. Abbott Park IL 60048-6202
Would anyone using a Nikon Coolpix 995 digital camera on a Nikon Eclipse series compound microscope please contact me regarding your experiences with this setup. I'd like to know the pro's and con's of working with this system to produce publishable images.
Mary, find someone with a focused ion beam (FIB) system that is willing to try non-semiconductor applications, otherwise it will take you just this side of forever to produce both plan view and cross section specimens. Failing that, you have a real challenge on your hands, I'm afraid. If you can make do with cross sections only, do a Listserver search on tripod polishing.
Tom Malis Materials Technology Lab Ottawa, Canada
} From: Ayten Celik {celik-at-students.uiuc.edu} } Date: Tue, 12 Feb 2002 15:19:18 -0600 (CST) } To: {Microscopy-at-sparc5.microscopy.com} } Subject: More detail..Re: Suggestions on sapphire samples } } ------------------------------------------------------------------------ } 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, } } I will be preparing both plane view and cross sectional samples. My main } concern is to keep the metal film in its initial state throughout the } sample preparation steps (as much as possible). The metal films are } generally 200-300 Angstrom thick. } } Can dimpler alter/damage the thin film? } } Second problem is the hardness of the sapphire. Do I have to use diamond } products to reduce the thickness of the sapphire substrates? The } substrates are 0.5 mm to start with. } } For grinding and polishing few different type of equipment are available } (from hand polising to automatic polishing machines). Getting access to } equipment is not the main concern. } } } Thanks } Ayten C. Aktas } } } On Mon, 11 Feb 2002, Ayten Celik 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 } } -----------------------------------------------------------------------. } } } } } } } } } } Dear all, } } } } I'm trying to prepare tem samples from sapphire. } } There is a thin metal film on the sapphire substrate, as well. } } I don't have too much experience in this field so, I would greatly } } appreciate any suggestions about preparing tem samples from sapphire. } } } } Previously, I have prepared couple of Si samples but, sapphire seems to be } } much harder and difficult to deal with. } } } } } } Thank you very much, } } Ayten C. Aktas. } } } } } }
I had a couple of people ask me about the PBL curriculum at the UH medical school as part of the thread about answering the questions from the students at Georgia Tech. For those who are interested, a statement about the philosophy can be found by visiting http://hawaiimed.hawaii.edu and clicking on the Medical Education link.
I'm not associated with the medical school, but the med students I've know have been very happy with it. However, I haven't had to visit any of them in an emergency, yet...!
Aloha, Tina
**************************************************************************** * 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* ****************************************************************************
I can only give a personal opinion based on my own experience with fluorescence microscopy. For low-light applications I personally prefer intensified cameras like the Isis-3 from Photonic Science ( http://www.photonic-science.ltd.uk/zzt_isis3.html ). These cameras allow you to work in extreme dim working conditions, which will keep your biological assay simple. You will not need to use several intensifying steps by using multiple layers of antibodies in an assay to get enough signal to get over the detection limit of your camera. These cameras will also allow you to keep up the speed of your image acquisition as you will not need to integrate the signal and thereby slow down the image capture process.
The main disadvantage of this approach is the relatively low S/N ratio in your images, but this can be compensated for with the proper digital filters and analysis algortihms.
When using cameras without image intensifier, you will probably need to integrate the signal over a certain period of time, which will slow down the acquisition proces and won't allow you to analyse "fast" dynamic processes. The benefit of this approach is a better S/N ratio than with intensified cameras and subsequent a more simplified analysis.
One of the issues under consideration is however the size of the CCD-elements on the camera, which are a measure of how much photons can be accumulated. There is an inverse relation between the size of the CCD-elements and the Nyquist sampling rate. For a given resolution (~ N.A.) of a microscope, you will need to magnify you image more if the CCD-elements are bigger than with smaller CCD-elements on the chip. You need to match the resolution of your microscope to that of your camera to obtain optimal results.
But there is also an inverse relation between the magnification and the amount of light (~I) hitting your camera ( I am not quite sure anymore, but it is something like this: I = N.A.^4 / M^2). So, the more you magnify your image, the more light you lose. Low-light fluorescence microscopy with immersion-oil lenses (high N.A.) and intensified cameras gives exciting results, but is sometimes a bit messy ;-)
Best regards,
Peter Van Osta
-- Dr. Peter Van Osta
Union Biometrica N.V./S.A. European Scientific Operations
Cipalstraat 3 B-2440 Geel Belgium
tel.: +32 (0)14 570 619 fax.: +32 (0)14 570 621
Michael Herron 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 } -----------------------------------------------------------------------. } } All, } } OK so it is a given that consumer grade cameras have relativly poor low } light performance. That said, are there cameras that have better than } average lowlight performance? Are any of the consumer cameras capable } of binning? } } Mike
Any local Kodak distributor should be able to order the 4489 film. It takes 2-4 weeks for delivery. They probably will not keep it in stock, however most places should be a able to order it. You could try Kodak's main office Rochester, NY for names and phone numbers of local distributors. Good Luck.
I am assessing vastly different systems that use the Nikon DN100, the Sony DXC390, or the Q-Imaging Micropublisher cameras for capturing bright field microscopy (mainly frozen biopsies). I am seeking any comments or experiences (benefits/limitations) about any of these three cameras (or others). For example, ease of use, software friendliness, quality, robustness, . . .
Please feel free to send responses offline, if they are not appropriate for the list.
Thank you,
Peter O. Steele, Ph.D. Pathology and Laboratory Medicine All Children's Hospital St. Petersburg, Fl
Randy, Of course it's too good to be true...for now. We are currently working on new ways to synthesize polypyrrole (and polyaniline, also a conducting polymer). Unfortunately, these and related conducting polymers are sensitive to temperature, and only conduct if doped properly (in certain pH ranges with certain dopants). Plus, the films themselves are quite brittle and somewhat difficult to manipulate. Like all new materials, though, it is a promising technology, and applications such as the one you mentioned is a definite possibility. File the provisonal patent now!
Paul E. Anderson Post-doctoral Research Associate Northeastern University Department of Chemistry 102 Hurtig Hall Boston, MA 02115
Gordon, I get my 4489 from VWR and Electron Microscopy Sciences. The Med. School contracts with VWR, so we get a bit of a discount from them, but sometimes they don't have it in stock. Lee -- Lee Cohen-Gould Electron & Optical Microscopy Facilities Weill Medical College of Cornell U. (212)746-6146 Rms A-105, LC-207
Short Course ?Specific localisation methods and microscopy in Food Research.?
Sunday May 5th, 2002, 8:00 am - 6:00 PM Electron Microscopy Centre, McGill University Montréal, Québec, Canada
Sponsored by the Food Structure & Functionality Forum Division of the AOCS
Short Course Organizer: Marcel Paques, Wageningen Centre for Food Sciences / Unilever R&D Vlaardingen, The Netherlands
Spreadability, shelve life, fracture behaviour, and creaminess are examples of functional properties of food products. These properties originate from the microscopic structure of products. Specific localisation techniques and microscopy are powerful tools to facilitate intelligent modification of ingredient composition or processing to obtain targeted food product properties. The short course is aimed at R&D personnel in the Foods area (fundamental research, innovation, and product development). The course consists of lectures and an intensive hands-on practical section providing participants with sufficient basic knowledge and skills to set-up and implement the methods in their own work. Registered participants are encouraged to submit application-related questions to the instructors by email prior to the short. In addition a personal consultation is offered to each participant scheduled (by appointment) during the AOCS Annual Symposium 2002 in the days directly following the short course. Contact Marcel Paques at paques-at-nizo.nl with questions.
Programme topics include:
· Pre-course consultation (by email) with participants to ensure the course content is relevant and applicable to participants? interests · Introduction to specific localisation methods and principles · Localisation strategies, marking options, and imaging approaches · Experimental set-up, preparation and incubation procedures · Demonstration examples · Hands-on practical sessions · Tailored help and advice during private consultation session following short course
Course contributors: Jan Leunissen (Aurion: immuno Gold Reagents & accessories, custom labeling, The Netherlands) Hong Yi (Emory Neurology Microscopy Core Laboratory, Emory University Department of Neurology, USA) Marcel Paques (Wageningen Centre for Food Sciences/Unilever Research Vlaardingen, The Netherlands)
Registration Fee: $375. The registration fee includes complete course materials, continental breakfast, lunch, two refreshment breaks, and transportation to and from McGill University.
Space is limited so register online today! http://www.aocs.org/meetings/am2002/fscourse.htm
This electronic message is sent by NIZO food research to its business partner and may contain confidential information only to be used by the client. The contents may not be used by, copied or revealed to any other person than the addressee. In case this message was mistakenly addressed to you, please return the message to info-at-nizo.nl or call +31 (0)318 659 511
If I'm not mistaken, National Graphic Supply in Albany carries this film in stock. In the past, George Laing has been our contact there for photographic supplies. The number I have for them is 1-800-223-7130, and George's extension is 3109. I haven't been in touch with them for awhile, but George has always been very helpful in the past.
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/
-----Original Message----- } From: Gordon Nord [mailto:gnord-at-mindspring.com] Sent: Tuesday, February 12, 2002 12:25 PM To: Microscopy-at-MSA.Microscopy.Com
Dear List,
We need a New York City source for Kodak Electron Microscope film 4489 (3.25" x 4").
Arkin Medeo was our previous source but their phones are disconnected and we can't find them.
Thanks, Gordon Nord
-- Gordon L. Nord Jr. Environmental Sciences Laboratory Brooklyn College
Ayten; I prepared some silicon on sapphire samples at Lockheed a number of years ago using dimple and ion mill techniques, but it is extremely challenging. The sapphire is nearly as hard as diamond, so the polishing rates are extremely slow. If you get aggressive, then it cracks. If you want to look at a metal film on top of the sapphire, you should cap it with SiO2, or it will be corroded/eroded and destroyed by the time the sapphire gets thin. I never tried plan views, but they should be similarly challenging. You will also find ion milling rates to be extremely slow, and must be done with the beam crossing the sapphire first, using rocking of the sample (never rotating) or the metal film will be lost. Tom Malik's suggestion of finding a FIB should be taken very seriously.
John Mardinly Intel
-----Original Message----- } From: Ayten Celik [mailto:celik-at-students.uiuc.edu] Sent: Tuesday, February 12, 2002 1:19 PM To: Microscopy-at-sparc5.microscopy.com
Hi,
I will be preparing both plane view and cross sectional samples. My main concern is to keep the metal film in its initial state throughout the sample preparation steps (as much as possible). The metal films are generally 200-300 Angstrom thick.
Can dimpler alter/damage the thin film?
Second problem is the hardness of the sapphire. Do I have to use diamond products to reduce the thickness of the sapphire substrates? The substrates are 0.5 mm to start with.
For grinding and polishing few different type of equipment are available (from hand polising to automatic polishing machines). Getting access to equipment is not the main concern.
Thanks Ayten C. Aktas
On Mon, 11 Feb 2002, Ayten Celik 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 } -----------------------------------------------------------------------. } } } } } Dear all, } } I'm trying to prepare tem samples from sapphire. } There is a thin metal film on the sapphire substrate, as well. } I don't have too much experience in this field so, I would greatly } appreciate any suggestions about preparing tem samples from sapphire. } } Previously, I have prepared couple of Si samples but, sapphire seems to be } much harder and difficult to deal with. } } } Thank you very much, } Ayten C. Aktas. } }
One possible way to bypass automatic adjustment with the coolpix 995 is to set it to manual operation,and turn off the awb (auto white balance).
Regards
Alfredo
Faerber Jacques {Jacques.Faerber-at-ipcms.u-strasbg.fr} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
__________________________________________________________________ Your favorite stores, helpful shopping tools and great gift ideas. Experience the convenience of buying online with Shop-at-Netscape! http://shopnow.netscape.com/
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Gordon, Arkin Medo changed their number. The new number for Arkin Medo is (718) 445-4000. Frank
At 01:24 PM 2/12/02 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am searching for references on measurement of oxide film thickness on metals by WDS in the SEM or TEM (as an alternative to XPS/Auger). Does anyone have advice on the "best" journal papers on this topic?
Sincerely, Paul Baggethun ================== Alcoa Technical Center Alcoa Center, PA 15069 (724) 337-1760 ==================
One of my former students developed a very nice technique to make both plan view and cross sectional samples based on the Tripod Polisher from South Bay Technology. It is a high angle polishing technique that works particularly well for samples where the film and the substrate have very different hardness and milling rate. With this technique you can also monitor the actual thickness of the thinned part of the sample. We have successfully used the technique to make samples of sapphire as well as Si (particularly good for this technique), LaAlO3, SiC, GaAs, SrTiO3, etc. The reference for the description of the technique is
"The Concept of High Angle Wedge Polishing and Thickness Monitoring in TEM Sample Preparation," Hao Li, and L. Salamanca-Riba, Ultramicroscopy 88, 171-8 (2001).
If you want to see TEM images of samples made with this technique let me know and I will e-mail you some off line.
Good luck,
Lourdes Salamanca-Riba
----- Original Message ----- } From: Ayten Celik {celik-at-students.uiuc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, February 12, 2002 4:19 PM
Hi Listers,
This appeal goes out to all the old timers out there who remember back when all pictures were created in the darkroom (I still think film and paper give a better picture and you techies out there can't change my mind ;-) ).
I have an old Densi-timer model PTM-4A made by Lektra Laboratories, Inc. It is a swell little thing that helps determine the exposure time for a print by using some sort of magic using a red light pointed at a medium gray tone of the negative and several knobs geared towards what grade of paper is being used.
Now I'm an old hand a printing and am used to just kind of knowing what time and what kind of paper to use to make a pretty picture, but my young helpers aren't. Does anyone know of a similar type device that is new & modern? This puppy has tubes and is on it's last legs.
I have money buring a hole in my pocket and Uncle Sam will take it back if I don't spend it soon. Any suggestions as to where to look and what's out there are greatly appreciated. Folks I talked to out here just gave me a blank stare.
Help keep me in the dark(room),
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
We are pleased to announce the availability of a Summer Internship at Intel Santa Clara for the Summer of 2002. The successful candidate will develop techniques for TEM Electron Tomographic 3D imaging of microelectronic structures. This work will involve everything from focused ion beam specimen preparation, imaging, reconstruction, rendering, and presentation via digital movies. The ideal candidate should be a graduate student in Materials Science, Physics, or equivalent, with experience in general theory and use of transmission electron microscopes, and be particularly facile with PCs and image processing. Unix/Linnux/SG experience would be a great asset. This work is being conducted in collaboration with the Agard Laboratories at UCSF, and frequent travel between Santa Clara and San Francisco will be required. Non-citizen candidates must have a Green Card or other legal right to work in the United States. Interested candidates should contact me and submit resumes as soon as possible either by e-mail, phone, or at the below mailing address. Thank you.
John Mardinly John.Mardinly-at-Intel.com Intel Corp. 2200 Mission College Blvd. SC9-7 Santa Clara, CA 95054 Desk: 408-765-2346 Pager: 408-322-6490
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hina-at-ohio.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, February 13, 2002 at 11:24:14 ---------------------------------------------------------------------------
Email: hina-at-ohio.edu Name: Sarah Hina
Organization: Ohio University
Education: Graduate College
Location: Athens, OH USA
Question: I work in a lab where we are investigating hair bundles of the utricle (inner ear). I have a question regarding osmolarity and TEM fixation. We are using the De Groot fixative, which contains 3% Glutaraldehyde, 2% Paraformaldehyde, 1% Acrolein, and 2.5% DMSO in 0.8M Sodium Cacodylate-HCL buffer. The total osmolarity is 1149 mmol/kg, considerably more hypertonic than the extracellular fluid (about 280 osmoles). I have read that glutaraldehyde and formaldehyde do not contribute significantly to the effective osmotic pressure of the fixative. Is this true? Does anyone know if acrolein and DMSO contribute to the effective pressure? We measured the osmolarity of our buffer alone, and it was a more reasonable 415 mmol/kg. Is this the more important value? Also, we are fixing by way of vascular perfusion, which according to what I have read, requires a more hyperosmolar solution. I would very much appreciate any comments or suggestions. Thanks!
Gordon, Although not in NYC, National Graphic Supply of Albany keep EM film in stock and can ship the same day of order. They give a special discount price for Colleges and Universities. Phone: 1-800-223-7130 or check their web site at http://www.ngscorp.com
Disclaimer: I have no commercial interest in the firm. They have supplier us satisfactorily for many years . Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences Binghamton University Binghamton, NY 13902-6000
-----Original Message----- } From: Gordon Nord [mailto:gnord-at-mindspring.com] Sent: Tuesday, February 12, 2002 1:25 PM To: Microscopy-at-MSA.Microscopy.Com
Dear List,
We need a New York City source for Kodak Electron Microscope film 4489 (3.25" x 4").
Arkin Medeo was our previous source but their phones are disconnected and we can't find them.
Thanks, Gordon Nord
-- Gordon L. Nord Jr. Environmental Sciences Laboratory Brooklyn College
I use an X-Rite densitometer for critical neg analysis and printing. Sorry, not TEM or SEM negs. But the trick is to collect the D of highlights and shadows and then determine the total D range of the neg. This then leads to the optimal contrast factor for print paper and how much dodging and burning is necessary for a final print.
If you scan the original neg at high rez, you can get a good idea of its range from PS histogram. Then, adjust accordingly.
gary g.
At 12:37 PM 2/13/2002, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Adaptive behaviour and non-linearity of the response of a camera are common problems in digital microscopy. Most of the cameras I prefer to use have the option to disable this adaptive behaviour, but also the framegrabber/digitizer can have an adaptive response. For quantitative microscopy I prefer to disable the adaptive behaviour on both the camera and the framegrabber.
In general it is best to calibrate the response of a camera and framegrabber with a "dark" image and a "white" image which corresponds with the dynamic range of the microscope image(s) you intend to acquire after disabling the adaptive behaviour. Afterwards you can use these images for calculation of the optimal dynamic range.
For JPEG and TIFF file formats, it is generally a bad idea to use JPEG compression for images that need to be analysed afterwards, certainly for color images as this will introduce artefacts. For B/W images, JPEG compression can be used, depending on the kind of analysis that follows or if the images are only meant for displaying and printing. In my opinion it is no use to acquire an image with a 100X high N.A. oil-immersion lens and afterwards destroy the fine detail with JPEG-compression. One of the advantages of JPEG-compression however is that there are hardware compression engines available, which allow you to do the compression in real-time which might be useful if speed is an issue.
For images that are meant to be analysed, I personally prefer lossles LZW-compression on TIFF images, certainly for color images. It gives you less data-compression than is possible with JPEG-compression, but the quality of the iamges is better preserved.
In general one needs to be aware of the difference between lossy and lossles compression algorithms and what the images are meant for in the end.
Best regards,
Peter
-- Dr. Peter Van Osta
Union Biometrica N.V./S.A. European Scientific Operations
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Peter:
I think your assessments have a lot to do with your requirements and your plans for the images. If you plan on doing spatial measurements, then you will perhaps reap added benefit from the larger resolution options with higher pixel counts. If you are doing color-segmentation-related measurements, then the extra color fidelity of the 3-CCD Sony might offer the ability to better distinguish similar colors, but at lower spatial resolution. If color fidelity is a requirement, but so is the high pixel resolution, you could cloud the issue even further by considering 3-CCD, mega-pixel options from Sony or JVC. (I can provide info offline if these options are of interest.)
If you have no plans for measurement, only archival or publication, then your assessment does fall back on the items you mentioned. I can't speak from experience to the ease of use or software friendliness of the Nikon or the Q-Imaging, but my guess is that both have their own software applications that acquire and save, and both probably offer TWAIN for use with Photoshop, etc. The Sony DXC-390 is an analog 3-CCD camera and does not provide for its own direct acquisition of images into the computer. This camera will have to be connected to a framegrabber and the ease of use and software friendliness will depend on what framegrabber board has been proposed with the camera. The image quality from this camera will also depend on the board. The best image quality from this camera will be the RGB output. If either the S-Video or the NTSC outputs are used for acquisition, the image will still be there (and the framegrabber might cost less), but it won't be the optimal image. Software support for the framegrabber board could be the manufacturer's application, or it could include some 3rd-party package that might even include image analysis.
In all cases, software ease of use will depend on what functions have been implemented into the software support. In some cases, although not all, cameras have capabilities that can be accessed through the user menus, or through Software Development Kit (SDK) programming calls, but those features are not always conveniently included in the free control application provided with the camera or framegrabber.
If I drone on much further, the relevance to the group may dwindle quickly, if it hasn't already. I hope this info helps a little, even though it's perhaps more conceptual, as opposed to directly related to your three options.
*Kind Regards, *Dave Hall *Resolution Technology, Inc - (614) 921-0045 *Please visit our website at http://www.restechimage.com
-----Original Message----- } From: Peter Steele [mailto:STEELEP-at-allkids.org] Sent: Wednesday, February 13, 2002 8:50 AM To: Microscopy-at-sparc5.microscopy.com
I am assessing vastly different systems that use the Nikon DN100, the Sony DXC390, or the Q-Imaging Micropublisher cameras for capturing bright field microscopy (mainly frozen biopsies). I am seeking any comments or experiences (benefits/limitations) about any of these three cameras (or others). For example, ease of use, software friendliness, quality, robustness, . . .
Please feel free to send responses offline, if they are not appropriate for the list.
Thank you,
Peter O. Steele, Ph.D. Pathology and Laboratory Medicine All Children's Hospital St. Petersburg, Fl
A colleague of mine and I were taking a look at a piece of this material and we are curious to know more about it. I know some suppliers carry vitreous carbon planchets for SEM, but we are curious to know exactly what they are .For example, is this material carbon that has been densified more than "amorphous" carbon due to higher pressures or perhaps the material has been "quenched" in some fashion or a combination of both ? Also, besides offering a much smoother surface (for SEM or AFM) do these planchets have higher conductivity than regular carbon planchets ?
} Email: hina-at-ohio.edu } Name: Sarah Hina } } Organization: Ohio University } } Education: Graduate College } } Location: Athens, OH USA } } Question: I work in a lab where we are investigating hair bundles of } the utricle (inner ear). I have a question regarding osmolarity and } TEM fixation. We are using the De Groot fixative, which contains 3% } Glutaraldehyde, 2% Paraformaldehyde, 1% Acrolein, and 2.5% DMSO in } 0.8M Sodium Cacodylate-HCL buffer. The total osmolarity is 1149 } mmol/kg, considerably more hypertonic than the extracellular fluid } (about 280 osmoles). I have read that glutaraldehyde and } formaldehyde do not contribute significantly to the effective osmotic } pressure of the fixative. Is this true? Does anyone know if } acrolein and DMSO contribute to the effective pressure? We measured } the osmolarity of our buffer alone, and it was a more reasonable 415 } mmol/kg. Is this the more important value? Also, we are fixing by } way of vascular perfusion, which according to what I have read, } requires a more hyperosmolar solution. I would very much appreciate } any comments or suggestions. Thanks! } } ---------------------------------------------------------------------------
Read Arborgh et al. J. Ultrastruc. Res. 56:339-350, 1976. A classic work. Basically they found that the osmolarity that mattered was the osmolarity of the buffer, not the buffer-fixative combo. Somewhat hypertonic was better than isotonic. As always, the proof of the pudding is in the eating. I am guessing that your cacodylate buffer is 0.08M, not 0.8M?
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 **********************************************
Afternoon Sara, This problem is quite special and most good information is derived from studies on specific tissues. The upshot is that one is best advised to perform one's own test starting with the information that is offered in good primary and secondary sources. I will recommend two secondary sources: Stoward, P.J., Fixation in Histochemistry", Chapman and Hall, London, 1973 (ISBN 412-12050-X) Hayat, M.A., "Fixation for electron Microscopy", AP, NY, NY, 1981 (ISBN: 0-12-333920-0) Hayat also has a more recent edition of his book entitled "Principles and Techniques in Electron Microscopy" that you might also want to consult. For more recent contributions that might be relevant, I suggest a search of PubMed at NCBI: http://www.ncbi.nlm.nih.gov/. I used "fixative osmolarity inner ear" and got some interesting looking results.
Regards,
Fred Monson
Frederick C. Monson, PhD The best research Center for Advanced Scientific Imaging occurs before work West Chester University at the bench. West Chester, Pennsylvania, USA, 19383 610-738-0437 fmonson-at-wcupa.edu
} ---------- } From: hina-at-ohio.edu } Sent: Wednesday, February 13, 2002 5:25 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Ask-A-Microscopist: osmolarity and TEM fixation } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (hina-at-ohio.edu) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } February 13, 2002 at 11:24:14 } -------------------------------------------------------------------------- } - } } Email: hina-at-ohio.edu } Name: Sarah Hina } } Organization: Ohio University } } Education: Graduate College } } Location: Athens, OH USA } } Question: I work in a lab where we are investigating hair bundles of } the utricle (inner ear). I have a question regarding osmolarity and } TEM fixation. We are using the De Groot fixative, which contains 3% } Glutaraldehyde, 2% Paraformaldehyde, 1% Acrolein, and 2.5% DMSO in } 0.8M Sodium Cacodylate-HCL buffer. The total osmolarity is 1149 } mmol/kg, considerably more hypertonic than the extracellular fluid } (about 280 osmoles). I have read that glutaraldehyde and } formaldehyde do not contribute significantly to the effective osmotic } pressure of the fixative. Is this true? Does anyone know if } acrolein and DMSO contribute to the effective pressure? We measured } the osmolarity of our buffer alone, and it was a more reasonable 415 } mmol/kg. Is this the more important value? Also, we are fixing by } way of vascular perfusion, which according to what I have read, } requires a more hyperosmolar solution. I would very much appreciate } any comments or suggestions. Thanks! } } -------------------------------------------------------------------------- } - } }
Thank you very much for your suggestion--I will be sure to check out that article. And yes, it should have read 0.08M. Thanks for your time--I appreciate it!
Sarah
--On Thursday, February 14, 2002 11:57 AM -0500 Geoff McAuliffe {mcauliff-at-UMDNJ.EDU} wrote:
} hina-at-ohio.edu wrote: } } } Email: hina-at-ohio.edu } } Name: Sarah Hina } } } } Organization: Ohio University } } } } Education: Graduate College } } } } Location: Athens, OH USA } } } } Question: I work in a lab where we are investigating hair bundles of } } the utricle (inner ear). I have a question regarding osmolarity and } } TEM fixation. We are using the De Groot fixative, which contains 3% } } Glutaraldehyde, 2% Paraformaldehyde, 1% Acrolein, and 2.5% DMSO in } } 0.8M Sodium Cacodylate-HCL buffer. The total osmolarity is 1149 } } mmol/kg, considerably more hypertonic than the extracellular fluid } } (about 280 osmoles). I have read that glutaraldehyde and } } formaldehyde do not contribute significantly to the effective osmotic } } pressure of the fixative. Is this true? Does anyone know if } } acrolein and DMSO contribute to the effective pressure? We measured } } the osmolarity of our buffer alone, and it was a more reasonable 415 } } mmol/kg. Is this the more important value? Also, we are fixing by } } way of vascular perfusion, which according to what I have read, } } requires a more hyperosmolar solution. I would very much appreciate } } any comments or suggestions. Thanks! } } } } ------------------------------------------------------------------------ } } --- } } Read Arborgh et al. J. Ultrastruc. Res. 56:339-350, 1976. A classic work. } Basically they found that the osmolarity that mattered was the osmolarity } of the buffer, not the buffer-fixative combo. Somewhat hypertonic was } better than isotonic. } As always, the proof of the pudding is in the eating. } I am guessing that your cacodylate buffer is 0.08M, not 0.8M? } } 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 } ********************************************** } }
Thanks so much for your suggestion--I did find a copy of the Hayat book, which provided some good info. I think the gist of their rationale is that the osmolarity of the buffer is more important than that of the fixtative. In our case, the acrolein and buffer would be first to enter the tissue, thus partially fixing it before the other fixatives could affect the tissue. However, I will be sure to conduct a search through pubmed as well. Thanks again for your time!
Sarah
--On Thursday, February 14, 2002 2:29 PM -0500 "Monson, Frederick C." {fmonson-at-wcupa.edu} wrote:
} Afternoon Sara, } This problem is quite special and most good information is derived } from studies on specific tissues. The upshot is that one is best advised } to perform one's own test starting with the information that is offered } in good primary and secondary sources. } I will recommend two secondary sources: } Stoward, P.J., Fixation in Histochemistry", Chapman and } Hall, London, 1973 (ISBN 412-12050-X) } Hayat, M.A., "Fixation for electron Microscopy", AP, NY, NY, } 1981 (ISBN: 0-12-333920-0) } Hayat also has a more recent edition of his book entitled } "Principles and Techniques in Electron Microscopy" that you might also } want to consult. } For more recent contributions that might be relevant, I suggest a } search of PubMed at NCBI: http://www.ncbi.nlm.nih.gov/. I used "fixative } osmolarity inner ear" and got some interesting looking results. } } Regards, } } Fred Monson } } Frederick C. Monson, PhD } The best research } Center for Advanced Scientific Imaging } occurs before work } West Chester University } at the bench. } West Chester, Pennsylvania, USA, 19383 } 610-738-0437 } fmonson-at-wcupa.edu } } } } ---------- } } From: hina-at-ohio.edu } } Sent: Wednesday, February 13, 2002 5:25 PM } } To: microscopy-at-sparc5.microscopy.com } } Subject: Ask-A-Microscopist: osmolarity and TEM fixation } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } } submitted by (hina-at-ohio.edu) from } } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } } February 13, 2002 at 11:24:14 } } ------------------------------------------------------------------------ } } -- - } } } } Email: hina-at-ohio.edu } } Name: Sarah Hina } } } } Organization: Ohio University } } } } Education: Graduate College } } } } Location: Athens, OH USA } } } } Question: I work in a lab where we are investigating hair bundles of } } the utricle (inner ear). I have a question regarding osmolarity and } } TEM fixation. We are using the De Groot fixative, which contains 3% } } Glutaraldehyde, 2% Paraformaldehyde, 1% Acrolein, and 2.5% DMSO in } } 0.8M Sodium Cacodylate-HCL buffer. The total osmolarity is 1149 } } mmol/kg, considerably more hypertonic than the extracellular fluid } } (about 280 osmoles). I have read that glutaraldehyde and } } formaldehyde do not contribute significantly to the effective osmotic } } pressure of the fixative. Is this true? Does anyone know if } } acrolein and DMSO contribute to the effective pressure? We measured } } the osmolarity of our buffer alone, and it was a more reasonable 415 } } mmol/kg. Is this the more important value? Also, we are fixing by } } way of vascular perfusion, which according to what I have read, } } requires a more hyperosmolar solution. I would very much appreciate } } any comments or suggestions. Thanks! } } } } ------------------------------------------------------------------------ } } -- - } } } }
Hello all: I am trying to find a/some references on methods for TEM of cellulose fibers. I have found what look to be good ones but they are in German. Can anyone direct me to something in English. Thanks in advance.
Stephen McCartney Research Associate 2108 Hahn Hall Materials Institute Virginia Tech Blacksburg, VA 24061-0344 USA
Does anyone know if there are any differences in how Technovit is used and works compared to Historesin.
I am a clinical pathology laboratory embedding human kidneys for diagnosis. I am used to how Historesin is processed/cuts/stains/etc. What can I expect with Technovit? The same? In Pathology, you never *experiment* with a patient's specimen/life...so I'm a little nervous in having to switch from one product to another.
Any insight/experience in this issue is appreciated.
Karen Bovard Electron Microscopy Laboratory Department of Pathology Creighton University/St. Joseph Hospital Omaha, Nebraska
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mp0017-at-unt.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, February 14, 2002 at 13:45:05 ---------------------------------------------------------------------------
Email: mp0017-at-unt.edu Name: M. Pritchett
Organization: University of North Texas
Education: Graduate College
Location: Denton, TX
Question: We are trying to get atomic resolution and find after spot welding the sample very well, it only seems to hold through two annealing cycles. (We anneal to 650K for 30 minutes to get the surface smooth.) It seems the repeat annealing loosens the sample because then we get lots of noise.
Is there some kind of adhsive that can be used to keep a Cu(1 1 1) sample from moving? Or is there another soution to our problem?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pdrum-at-island.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, February 14, 2002 at 10:38:09 ---------------------------------------------------------------------------
Email: pdrum-at-island.net Name: Peter Drummond, Ph.D.
Organization: North Island College
Education: Undergraduate College
Location: Port Alberni, British Columbia
Question: How in most junior colleges, are the inventory of microscopes maintained in working order? And Would you know if they are any microscope maintenance programs available in the next... say 6 months? Thanks.
Please put your thinking caps on and help me with advice for a researcher here with an interesting problem.
The researcher in question has made 100 nm pores in mica which he fills with polyacrylamide gel. He would like to visualize the pores and see if they are filled with gel, or if not, what the proportion of gel to space is in the pore.
He vetoed our SEM because the gel will shrink too much for him to see what is going on. I suggested an ESEM but don't know if that would really work.
I thought of doing some kind of fluorescent labels and then maybe something like a confocal but he says he hasn't found a dye that will work for that approach.
He can make the pores larger, up to a micrometer or so, but 100 nm is his nominal best size for other reasons. If you have any ideas or practical experience with this kind of problem, please let me know and I will forward your words of wisdom to him.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
AFM in Tapping mode with phase imaging may be able to visualise this. The phase imaging will be able to tell between the mica and the polyacrylamide. This may, however be practically difficult as the acrylamide may cause problems with the tip.
DeltaDOT Ltd. PFSG group ACE Building Department of Bioengineering Imperial College London SW7 2AY
0207-594-5174 0794 - 1312335
www.deltadot.com
Information on scanning probe & microfluidics : www.achem.ic.ac.uk/gsanders **************************************************************************** ******* **************************************************************** ----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, February 15, 2002 12:29 AM
Karen,
I'm certain you will find the Technovit H7100 kit to be identical to Leica Historesin in all facets. However, if someone on the list has a different opinion I would be very eager to hear it. My knowledge comes not so much from use but from my knowledge of the source for and contents of both kits.
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 "Adding Brilliance to Your Vision"
Disclaimer: Energy Beam Sciences, Inc. is the North American distributor for Heraeus Kulzer "Technovit" Products.
-----Original Message----- } From: "kbovard-at-creighton.edu"-at-sparc5.microscopy.com [mailto:"kbovard-at-creighton.edu"-at-sparc5.microscopy.com] Sent: Thursday, February 14, 2002 4:57 PM To: Michiel De Mol Cc: Microscopy Listserver
Does anyone know if there are any differences in how Technovit is used and works compared to Historesin.
I am a clinical pathology laboratory embedding human kidneys for diagnosis. I am used to how Historesin is processed/cuts/stains/etc. What can I expect with Technovit? The same? In Pathology, you never *experiment* with a patient's specimen/life...so I'm a little nervous in having to switch from one product to another.
Any insight/experience in this issue is appreciated.
Karen Bovard Electron Microscopy Laboratory Department of Pathology Creighton University/St. Joseph Hospital Omaha, Nebraska
I've used both (Technovit 7100 and Historesin; also JB-4 and have made my GMA from the components). I use Technovit 7100 routinely and have found that it is very similar (if not identical) to Historesin. I use the standard recipe that comes with the package. You can purchase Technovit 7100 from Energy Beam Sciences. I just bought some and they have it in stock.
De Wood
At 03:56 PM 2/14/2002 -0600, kbovard-at-creighton.edu"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all who repsonded to my inquiry about my densi-timer. There were a lot of varied replies and a lot of folks still like to use film over digital.
Now for the next question....
I have 2 Zeiss EM-10's they both have attached to them a monitor that counts down the number of negatives left in the camera, tells the filament hours used and also tells a pre-vac pressure. One of them has now gone belly up when it comes to counting down the negatives left. Now I know that we could just do it the old fashioned way and count down using our hands and feet, but no one in the lab has 23 digits.
The company that made it was FBN Electronics, the number in New Jersey now belongs to a family. The model # is 10-4.
If any of you good buddies know anything about how to contact this company or knows if they just don't exist anymore, please let me know.
That's a big 10-4 good buddy,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Hi Jonathan, I think your suggestion of using an ESEM is spot on. Do you have access to one? It would provide you an opportunity to examine the material without a coating layer, and with minimal impact on the gel. Possibly a BSE detector might provide some enhanced contrast, but you'd have to test that. Good luck.
-Brad
-----Original Message----- } From: jmkrupp-at-cats.ucsc.edu [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Thursday, February 14, 2002 4:30 PM To: Microscopy-at-sparc5.microscopy.com
Greetings:
Please put your thinking caps on and help me with advice for a researcher here with an interesting problem.
The researcher in question has made 100 nm pores in mica which he fills with polyacrylamide gel. He would like to visualize the pores and see if they are filled with gel, or if not, what the proportion of gel to space is in the pore.
He vetoed our SEM because the gel will shrink too much for him to see what is going on. I suggested an ESEM but don't know if that would really work.
I thought of doing some kind of fluorescent labels and then maybe something like a confocal but he says he hasn't found a dye that will work for that approach.
He can make the pores larger, up to a micrometer or so, but 100 nm is his nominal best size for other reasons. If you have any ideas or practical experience with this kind of problem, please let me know and I will forward your words of wisdom to him.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Does anyone know of metallographic preparation techniques to produce cross-section specimens of bismuth-telluride (Bi2Te3) materials for thermolectric device applications. These are either single crystal or powder metallurgy materials. I wish to observe the cross-sections via light microscopy and SEM. Thanks in advance.
Photoshop is a widely used program for the acquisition, processing, annotation and printing of scientific images, and is frequently discussed on this list. It includes a custom convolution filter that allows users to enter a 5x5 array of integer weights that are multiplied by pixel values to produce smoothing, derivatives, high pass filters, and other useful effects. The interactive preview gives immediate feedback on the results, which is a great assist to learning about convolution kernels in general, and the operation can also be scripted in Actions for one-button application to images or for batch processing.
Unfortunately, the Photoshop “Custom†filter has a few significant limitations: 1. only works with 8 bit per channel grey and RGB images 2. applies the kernels separately to the RGB channels 3. is limited to a 5x5 array of integer coefficients, with integer scaling 4. saves and loads files in a special binary format
We’ve written and are offering for free download and use a very much enhanced version of this “custom†filter. Like the Photoshop one, it has an interactive preview, and is recordable in Actions. In addition, the plug-in: 1. supports both 8 and 16 bit per channel grey and RGB images 2. works on the intensity channel leaving hue and saturation unchanged 3. accepts up to a 7x7 array of floating point (real number) values, for much greater precision 4. can scale the results with a floating point number, or automatically for maximum unclipped contrast 5. reads Photoshop format files AND ALSO plain ascii text files 6. works in Photoshop and all compatible programs on both Mac and Windows computers
Encouraged by the recent response by readers of this list to our offering a free plug-in to draw magnification bars on micrographs, we are offering this plug-in for free download and use. It is one of nearly 200 plug-ins in the widely used Fovea Pro package, but can be used without installing or owning that package.
The plug-in, along with instructions and some example filter files, can be downloaded as a .sit file for Macintosh or a .zip file for Windows from {http://www.reindeergraphics.com/free.html#custom} . While you are there, please take a look at the full range of Fovea Pro capabilities, which include comprehensive tools for image processing and analysis. Also, check out some of the other free downloads that are available.
During the last week, several people have emailed me that they have tried to access the site with information on our short course but were unable to because of a server problem, which is now (apparently, hopefully, finally) fixed. Thanks for your patience. The May course is filling up but there are still places available. If you are interested in learning practical methods for the processing and measurement of images, check it out and sign up now.
The three-day intensive hands-on workshop on Image Processing and Measurement presented by John Russ (author of "The Image Processing Handbook" and "Practical Stereology") through the North Carolina State University Department of Continuing and Professional Education is now in its 20th year. The course dates for 2002 are May 8 - 10 in Raleigh, NC; June 10-12 at the Danish Technological Institute in Taastrup, Denmark (near Copenhagen); and November 6-8, 2002, in Raleigh. This course has generated highly favorable reviews from the thousands of previous students. The primary focus is on images from various types of microscopy, with practical guidance in correcting imaging defects, enhancing the images for presentation and measurement, and performing stereological meaningful measurements on them. Textbooks and computer software are provided to attendees. Lab sessions with an opportunity to bring your own images makes this course immediately useful and highly productive.
For full information on the course, including outlines, faculty information, a downloadable brochure, and on-line registration, go to
http://members.aol.com/ipcourse
Class size is limited to maintain a high ratio of instructors to students, so make your reservation now. You may also contact Cindy Allen at NCSU Continuing Education, at 919-515-8171
The safety and family planning problem occurs in light microscopy also.
Immersion oil may contain dibutyl phthalate (DBT), which can harm a developing fetus and also cause temporary male sterility.
Even though DBT is ubiquitous (in rocket fuel, paint thinner, fingernail polish, et al.,) and even though, so far as I know, there have been no studies on its absorption through the skin, its very, very, VERY high concentrations in some oils (hundreds or even thousands of times higher than OSHA standards for its presence in food, water or air), and the fact that most of our users are in their child-bearing years, seem to merit some concern. We now use a DBT-free oil, and ask our users to wash their hands before leaving the lab. (After all, before DBT, immersion oil contained PCBs!)
Joanne H. Whallon, Ph.D., Professor Department of Crop and Soil Sciences and Center for Advanced Microscopy Michigan State University East Lansing, MI 48824-1325
Phone 517/353-0837 or 517/432-2328 Fax: 517/353-3955 E-mail: whallon-at-pilot.msu.edu
It is not fair that the student does not have any chance to respond.
If I were that student, here is my respond:
You may think that giving me straight answer seems too easy on me. But on my side, it is not that simple to use your answer. If I ask my professor directly to get an answer, as long as I can repeat his/her words like 3 years old, I can get a A, even if he is WRONG.
But asking you is completely different. You could be WRONG too! If I took your wrong answer straight to my professor, I will fail. No matter how I explain this is from some "expert", it is not going to help.
Oh, how could YOU be wrong? Isn't that very insulting since you are the expert? If you felt that way, I am sorry. I did not mean to insult any one. However, how about post your answer to these "elementary" questions on this list server? Do you think everyone on this list will agree with you?
Here are the re-post of Jenny Wang's original question: 1. What are the advantages and disadvantages in using electrons for microscopy rather than light? 2. Does the wavelength of the electrons have anything to do with the spatial resolution that the microscope produces in the final picture? 3. What is temporal resolution and how is it produced in the electron microscope?
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Cryo-SEM would work well for this problem. The fast freezing would prevent shrinkage of the gel. In addition, if there is a thin surface layer of water it can be sublimated off to reveal the underlying surface. Since cryo-microscopes usually incorporate a coater, the sample could be coated and then viewed while frozen. If you are lucky enough to find a FEG-SEM with cryo than you may be able to work at low kV without coating. The sample could be removed, thawed, and used for another purpose if desired (with the understanding that the freeze-thaw could change the properties of the sample). We have done a lot of work with cryo-SEM and hydrated gels and it works great. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907
On Friday, February 15, 2002 2:06 PM, Johnson, Bradley R {Bradley.Johnson-at-pnl.gov} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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For most people, exposure to dibutyl phthalate in its role as a vinyl plasiticizer is perhaps more relevant than any amount that might be present in rocket fuel. (Babies, as well, seem more likely to chew on toys than the on the occasional chunk of propellant left lying around.)
If my memory is correct, phthalate plasticizer contents in soft vinyl have sometimes run up to about 20% by weight. Such plastics have been used in many large household items and probably account for a much more important route of exposure than immersion oils - even for sloppy microscopists. Vinyl's tendency to soften and cause printing ink and photocopy toner to transfer to adjacent surfaces is due to plasticizer migration. The loss of plasiticizer due to its volatilization and "sweating" gives rise to shrinkage and cracking in such plastics. In years past, who hasn't gotten into a hot car and smelled the bland but noticeable odor?
On the other hand, in 25 years of microscopy, I can't recall getting more than a milligram or two of immersion oil (PCB free!) on my skin. I guess that depends on one's technique.
While the jury may still be out on the full range of dibutyl phthalate effects, one who is concerned would want to deal with the major routes of exposure first. Perhaps you could expand on your other steps for the list.
What is the composition of your phthalate-free immersion oil?
Your last comment seems to invite the conclusion that immersion oils are inherently suspect, or that you believe phthalates will be found to pose risks similar to PCBs. Is that the case?
John Twilley
Joanne Whallon 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, List Folks: } } The safety and family planning problem occurs in light microscopy also. } } Immersion oil may contain dibutyl phthalate (DBT), which can harm a } developing fetus and also cause temporary male sterility. } } Even though DBT is ubiquitous (in rocket fuel, paint thinner, fingernail } polish, et al.,) and even though, so far as I know, there have been no } studies on its absorption through the skin, its very, very, VERY high } concentrations in some oils (hundreds or even thousands of times higher } than OSHA standards for its presence in food, water or air), and the fact } that most of our users are in their child-bearing years, seem to merit some } concern. We now use a DBT-free oil, and ask our users to wash their hands } before leaving the lab. (After all, before DBT, immersion oil contained } PCBs!) } } } } Joanne H. Whallon, Ph.D., Professor } Department of Crop and Soil Sciences and } Center for Advanced Microscopy } Michigan State University } East Lansing, MI 48824-1325 } } Phone 517/353-0837 or 517/432-2328 } Fax: 517/353-3955 } E-mail: whallon-at-pilot.msu.edu } } } }
My Zeiss Axiophot microscope has a recurring problem and I am wondering if any other users have the same problem and if they have come up with a permanent solution. The bright field lamp sits in a ceramic base and with time, the heat apparently cracks the base. As this is slowly happening, the lamp is no longer sitting firmly in its socket so the light fluctuates constantly. You can see it "breathing" as you watch the live image captured with a digital camera. I have replaced the ceramic socket at least 3 times and need to once again. Zeiss has not offered any solution other than to sell me a new socket each time. Any suggestions would be greatly appreciated.
-- 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 have waited for a week now and there don't seem to be any new responses to the "poll". Thanks to anybody who answered. As promised, I took a look at the numbers and here are the results. I am not going to interpret the numbers, you can do that for yourself.
The choices were:
} 1) Students should work through their assignments alone. Let's not support } laziness at all. } 2) We should help them help themselves by pointing them to general } microscopy books. } 3) We should take their questions and point them to specific microscopy } books dealing with their problems } 4) We should encourage them to contact us offline so we can test their } sincerity and then either help them or not. This should be posted on the } server to avoid redundancy. } 5) We should help them with their questions by trying to answer them in a } general way } 6) We should go all out and answer their questions as best as we can.
In total I counted 51 responses both to the list server and my email. I took out the double answers that were sent to both the list server and to me. Of the 51 responses, 25 were from educational facilities, 18 from commercial facilities, and 8 were from other sources (government, AOL, foreign). These classification was done on the email extension. A number of people selected more than one answer. In these cases I simply split up their single vote into partial votes. for example, if someone said "2,3, and 6", each of the choices got 0.33 votes. Finally, a couple of people thought, the selection of choices was too narrow and made other suggestions or withheld, which I put into number "7".
My intention was at no time to make this an exhaustive scientific article. I kept the choices simple and I only want to present the numbers in a simple way here. If you want to discuss the numbers, please do so, I will draw no conclusions from the numbers presented here.
Overall the results are as follows:
1 2 3 4 5 6 7 {- selection 1.2 9.53 23.03 4.03 4.7 6.5 2 {- number of votes
for .com 1 2 3 4 5 6 7 {- selection 0.2 2.7 9.2 2.2 1.7 2 0 {- number of votes
for .edu 1 2 3 4 5 6 7 {- selection 1 6.83 5.83 1.83 3 4.5 2 {- number of votes
for other 1 2 3 4 5 6 7 {- selection 0 0 8 0 0 0 0 {- number of votes
Unfortunately I can't send the Excel datasheet I used to tally the votes and show the numbers to the list server. If you are interested, drop me a line and I will send it to you.
mike
} } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Ave #300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
I had similar, but not identical problems, with my Axioskop and Axioplan lamp houses. Generally, they were quite good. If the lamps are run at full voltage (12VAC), they will get really hot and not last all that long. Also, as was recently discussed, line voltage is not constant. The Zeiss lamp controllers are simple triac controls which are not regulated. So, if your line voltage fluctuates, so will your lamp. that is the way it is.
I ran my halogen lamps at 10VDC using a Lambda regulated DC power supply. I also put a 12VDC 60mm fan next to the lamp housing. It turned out that just lowering the voltage made a huge difference. No fluctuation and long lamp life. The Axioplan stand seemed to have a regulated lamp source and would easily accommodate a 10V setting. I use HAL 100W 12V halogen bulbs (Osram). The same principle applies to any other wattage rating bulb. I solved this voltage fluctuation problem by converting all UPS units to dual conversion. My line logs show minimum line voltages of 103VAC to peak voltages of 118VAC. The UPS puts out a constant 120VAC....+- 3%. Nailed.... Yes, the units are a bit more expensive than passive UPS, but I don't have to deal with the hassle any longer. Even my unregulated Fostec ACE units work perfectly now and I don't have to buy DCR units at twice the price. I think this is a win situation.
Try using a lower voltage setting and see if that works for you. If not, put a cooling fan next to a vent port on the lamp house. Put it close to, but not on, the house. Otherwise, you will get vibration interference at high mag.
Contact me off-line if you need further info.
gary g.
At 11:15 AM 2/17/2002, you wrote:
} My Zeiss Axiophot microscope has a recurring problem and I am wondering if } any other users have the same problem and if they have come up with a } permanent solution. The bright field lamp sits in a ceramic base and with } time, the heat apparently cracks the base. As this is slowly happening, } the lamp is no longer sitting firmly in its socket so the light fluctuates } constantly. You can see it "breathing" as you watch the live image } captured with a digital camera. I have replaced the ceramic socket at } least 3 times and need to once again. Zeiss has not offered any solution } other than to sell me a new socket each time. Any suggestions would be } greatly appreciated. } } -- } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility
I have just produced a Catalogue of Portable Microscopes on CD-ROM. It lists approximately 280 instruments with 99% of them illustrated. It is ideal for microscope collectors and scientific instrument museums etc. For more information go to http://www.pnc.com.au/~dingley
Mike Dingley (I have a personal, financial and commercial interest in the CD).
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Does anyone know of a way to detect beta-galactosidase by TEM? The B-gal is expressed in transgenic mice. We would like to see it in the fibroblasts of nerve tissue. So far our literature search has yielded information at the light microscopy level only. Any ideas out there?
Thanks!
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
We currently have a need for ongoing EM-based (SEM and/or TEM) imaging and chemical analysis on a hourly or contract basis and full-service or instrument-only arrangements.
AMC Group is a consultancy based in New Mexico involved in physical and chemical characterization of advanced optoelectronic materials and devices. I would appreciate receiving responses from the intersested parties in US at both commercial labs and university labs off-line.
James Glossinger AMC Group, Inc. amcgroup2-at-aol.com
Hi At least two papers did B-gal enzyme cytochemistry at EM level. See Sekerkova
G. et al J Histochem Cytochem 45:1147-55, 1997 and Weis J. et al. JCB 113:1385-97, 1991. The method works well.
G. Ning Electron Microscopy Facility Medical College of Wisconsin
Dorothy Roak Sorenson 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 } -----------------------------------------------------------------------. } } Does anyone know of a way to detect beta-galactosidase by TEM? The B-gal } is expressed in transgenic mice. We would like to see it in the } fibroblasts of nerve tissue. So far our literature search has yielded } information at the light microscopy level only. Any ideas out there? } } Thanks! } } 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
Food Structure & Functionality Forum Symposium 2002 May 5 to 8, 2002, Palais des Congrès de Montréal, Montréal, Quebec, Canada
Monday, May 6th-Morning Opening of symposium
Presentation of Division Achievement Award to Dr. R. Gary Fulcher, University of Minnesota
Plenary address by R. Gary Fulcher: Title to be announced
Dairy Applications Session Chairs: Mark Auty, Dairy Products Research Centre, TEAGASC, Ireland (mauty-at-moorepark.teagasc.ie ) and Harjinder Singh, Massey University, NZ (H.Singh-at-massey.ac.nz)
Manufacturing Yoghurt Structures with a Predicted Consumer Preference. M Langton and A. Astrom, SIK, Sweden
Some Structure-Function Relationships in Milk Gels. J. Lucey, University of Wisconson-Madison, USA
Understanding the Functionality of Whey Proteins as Microencapsulating Agents. M. Rosenberg and M.H. Wang, University of California - Davis, USA
Carry-over Effects of Processing Parameters as Determined During Sweetened Condensed Milk Manufacture and Application. C. Attapattu, University of Wisconsin, USA and Y. Kakuda, University of Guelph, Canada.
Modifying Dairy Protein Functionality Through Extrusion Processing. C. I. Onwulata and R. P. Konstance, USDA-ARS-ERRC, USA
Some Observations of a Microscopist, Author, and Reviewer on Structural Studies of Milk Products. M. Kalab, Agriculture and Agri-Food Canada, Canada (Keynote Speaker)
Monday, May 6th - Afternoon (1:00-4:00PM Note Early Start) Colloidal and Interfacial Sciences Session. Chairs: Marcel Paques (Paques-at-nizo.nl ) and David Pechak (Dpechak-at-kraft.com)
rotein Polysaccharide Interactions. C.G. De Kruif, NIZO Food Research, Netherlands (Keynote Speaker
Influence of Protein Adsorption Conditions upon Oil-in-Water Emulsion Stability: A Comparison of b-Lactoglobulin to a Whey Protein Concentrate. B. Campbell1, I. Ivanon2, N. Denkov2, S. Tcholakova2, 1Kraft Foods,USA; 2University of Sofia, Bulgaria
Microstructure of Heat Processed Whey Protein Food Emulsion and Growth of Shear-Induced Cracks During Cooling. R. Ofstad1, V. Hoest1, O. Langsrud1, G. Enersen1, T.E. Nyvold2, E.P. Willers3, B. Nordvi4, B. Egelandsdal1,5, 1 Norwegian Food Research Institute-MATFORSK, Norway; 2Stabburet, Norway; 3Procordia Foods, Sweden; 4TINE Norwegian Dairies, Norway; 5Institute of Food Research, Agricultural University of Norway, Norway
Fatty Acid Salts-Induced Gels of Food Proteins: Their Rheological Properties and Structural Changes of the Proteins During Gelation. N. Yuno-Ohta, Nihon University, Japan
Wheat Gluten Proteins. A.S. Tatham, IACR Long Ashton Research Station, United Kingdom
nterfacial Composition and Stability of Oil in Water Emulsion formed with Mixtures of Milk Proteins and Polysaccharides. H. Singh and Y. Hemar, Institute of Food, Nutrition and Human Health, Massey University, NZ
Dedicated Poster Session
Division Board Meeting
Tuesday, May 7th - Morning (9AM-12PM) Agricultural Applications of Microscopy and Imaging Session/ joint with Feed Microscopy Division. Topic: Food Contamination contacts: Mark Auty, Dairy Products Research Centre, TEAGASC (mauty-at-moorepark.teagasc.ie ) and Kim Koch, North Dakota State University (kkoch-at-ndsuext.nodak.edu) (Feed Microscopy Division)
TBA. J. Shane, McCrone Institute.
How to approach contaminant identification. M. Auty, Dairy Products Research Centre, Ireland
Identification of plant material. D.F. Wood, USDA, USA
310 Species identification of Animal Hair by Using Atomic Force Microscopy. C.W. Cruywagen, University of Stellenbosch, South Africa.
401 Detection and Differentiation of APRALAN, PAYLEAN, PULMOTIL and TYLAN in Animal Feeds using microscopy. P. Klink. Elanco Animal Health, a Division of Eli Lilly and Company, USA.
Quantitation of Total Fat and Fat Quality in Cheese and Dairy Products Using Membrane Separation Technology. V.C. Gordon, Safety Associates, Inc., USA
Division Luncheon and round table (expert) discussion (12-2PM). Topic: Structure - functionality relationships in materials containing fats and oils: Drafting a Roadmap for the Future.
Specifics: Microstructure, Crystal Structure and Molecular Structure: Imaging, Quantification, and Relationships to Macroscopic Functionality - what is the state of the art, which relationships continue to baffle us, what are the promising new technologies/characterization methods/modeling activities?
Discussion leaders: M. Auty, M. Paques ( Food Structure and Functionality Forum) and S. Narine, N. Widlak (Edible Applications Division) *--------------------------------------------------------------------------------
Tuesday, May 7th - Afternoon ( Microbiology and Food Session Chairpersons: Judy Arnold and Ida Yates, USDA, ARS, Russell Research Center, USA
Prevention of Bacterial Fouling on Food Equipment Surfaces. J.W. Arnold, USDA, ARS, Russell Research Center, USA (
TBA. S. Pao, Virginia State University, USA
Yogurt microstructure and texture: The role of exocellular polysaccharides produced by lactic acid bacteria. J. F. Frank and A.N. Hassan. University of Georgia, USA
Probiotics and Their Use in Food Animal Production. R. Droleskey, USDA, ARS, SPARC, USA
Ultra-structural analysis of the monospecies biofilms formed by Listeria monocytogenes. M.L. Kalmokoff and J.W. Austin; BMH, Health Canada, Canada
The Effect of High Pressure Sterilization on Listeria Inoculated Seafood. K.R.S. Schneider and M.V.W. Wood, University of Florida, USA
Food Microstructure Investigations by Atomic Force Microscopy. J. Thornton, Digital Instruments/Veeco Metrology Group, USA (20 min)
Controlling Growth of the Toxigenic Fungus, Fusarium Verticillioides. I. Yates and J. Arnold, Russell Research Center, ARS, USDA, USA
Division Members Meeting (immediately following the afternoon session)
Wednesday, May 8th- Morning (8AM-12PM) Ingredients and Food Processing Session- Chairpersons: Diana Kittleson, General Mills Technology East, USA; and Bernhard Tauscher, Federal Research Center for Nutrition, Germany
Applications of Food Material Science. D.W. Stanley, University of Guelph, Canada (Keynote Speaker )
Non-Invasive Quality Determination of Fruit and Vegetables: Application of a Multi- Wavelength NIR-Diode Laser Array. B. Tauscher, P. Butz, Federal Research Center for Nutrition, Germany
Microstructure Characterization of Polysaccharide Based Films. G. Frias2, M.A. Garcia1, M. Martino1,2. 1CIDCA, UNLP, CONICET, Argentina, 2 Faculty of Engineering, UNLP, Argentina
Characterisation of the Application of Novel Oil Structurants. E. Floeter, F. Gandolfo, and W. Hogervorst, Unilever Research Vlaardingen, The Netherlands
Fat Bloom Formation and Characterization in Milk Chocolate Observed by Atomic Force Microscopy. S.M.Hodge, D. Rousseau, Ryerson University, Canada
High Pressure Processing. E. Ting and E. Raghubeer, Flow International, USA
Microstructure of Rice Starch Isolates. D.F. Wood1, A.M. Ibanez_Carranza2, and C.F. Shoemaker2, 1USDA, ARS, WRRC, USA; 2University of California, USA
Structural Properties of Multiphase Potato Products. M. Lamberti,F. Escher, B. Conde-Petit, ETH, Switzerland
Effect of Processing on Microstructure of Oat Starch. M. Salmenkallio-Marttila , K. Autio, VTT Biotechnology, Finland
Novel Techniques for the Production of Functional, Less Beany-Flavored Soy Products J. I. Boye, Agriculture and Agri-Food Canada, Canada. (
Wednesday, May 8th - Afternoon (2-5PM) New Methods and Techniques for Food Structure and Functionality Analysis Session Chairpersons: Kathy Groves, Leatherhead Food Research Association, England; and Maud Langton, SIK, Sweden
Quantifying Microstructures through Image Analysis. G.M.P. van Kempen1, M. van Ginkel2, C.L. Luengo Hendriks2, L.J. van Vliet2, and S. Singleton3, 1Unilever Research Vlaardingen, Netherlands; 2Delft University of Technology, The Netherlands; 3Unilever Research Colworth House, Great Britain (Keynote Speaker )
Changes in plant tissue after pulsed electric field treatment. M. Fincan, P. Dejmek Dept. of Food Engineering, Lund University, Sweden
Cryo-TEM of Biopolymers in Comparision with Other TEM-techniques. M. Langton, A. Altskar, and A.-M Hermansson, SIK, Sweden
Freeze-substitution and low temperature embedding of dairy products for electron microscopy. A.K. Smith and H.D. Goff, Department of Food Science, University of Guelph, Canada
MicroRheology: preliminary results of structural behaviour of foods under deformation. M. Paques, Y. Nicolas, Wageningen Centre for Food Sciences/Unilever Vlaardingen, The Netherlands.
Recent Advances in our Understanding of the Relationship Between Crystallization Behavior, Microstructure and Rheological Properties of Fat Crystal Networks. A. Marangoni, University of Guelph, Canada
CLOSURE OF SYMPOSIUM
Posters Spectroscopic Prediction of Rheological Properties in Grains. F. Meadows and F. Barton USDA, ARS, QARU, USA
Staining Techniques for Detection of Components in Fish Muscle. K. Hanneson Eggen and G. Enersen, Matforsk, Norway
Effect of Emulsifiers and Processing Conditions on Microstructure of Milk Fat/Sunflower Oil Blends. S. Martini1, M. Cerdeira1, C. Puppo1, R.W. Hartel2, and M.L. Herrera1,3, 1CIDCA, UNLP, CONICET, Argentina; 1University of Wisconsin-Madison, USA; 3University of Buenos Aires, Argentina
Exchange in Semi-Solid Triglyceride systems measured by NMR Spectroscopy: Effect of Partial Glycerides on Exchange Rates. P. Smith1, N. Haghshenas1, I. Furo2, and B. Bergenstahl3, 1YKI Institute for Surface Chemistry, Sweden; 2Royal Institute, Sweden; 3Lund University, Sweden.
Utilizing Polarized Light Microscopy to Characterize the Effects of Tween60 on the Physical Properties of a Model Plastic Fat System. J.W. Litwinenko and A.G. Marangoni, University of Guelph, Canada.
Relationships between Microstructure and Rheological Properties of Model Lipid Systems. B. Liang, Y. Shi, and R.W., Hartel Univ. of Wisconsin-Madison, USA
Texturization of Water-in-oil-Based Spreads. Y. Shi, B. Liang, and R.W. Hartel, University of Wisconsin-Madison, USA
Effect of Temperature and Shear Rate on Fat Crystallization Kinetics. Ph Rousset, C. Bertoli, H. Dux, Nestle Research Center, Switzerland
Formation and Physical Properties of ?-Fat Gel. I: Macroscopic and Microscopic Observations. K. Higaki1, Y. Sasakura1, I. Hachiya1, and K. Sato2, 1Meiji Seika Kaisha Ltd., Japan; 2Faculty of Applied Biological Science, Hiroshima University, Japan
Formation and Physical Properties of ?-Fat Gel. Ii: In situ Observation of Gel-Formation Processes. K. Higaki1, Y. Sasakura1, I. Hachiya1, S. Ueno2, and K. Sato2, 1Meiji Seika Kaisha Ltd., Japan; 2Faculty of Applied Biological Science, Hiroshima University, Japan
Formation and Physical Properties of ?-Fat Gel. III: Rheological Properties. K. Higaki1, T. Koyano1, I. Hachiya1, and K. Sato2, 1Meiji Seika Kaisha Ltd., Japan; 2Faculty of Applied Biological Science, Hiroshima University, Japan
Formation and Physical Properties of ?-Fat Gel. IV: Why and How? K. Sato1, K. Higaki2, T. Koyano2, and I. Hachiya2, 1Faculty of Applied Biological Science, Hiroshima University, Japan; 2Meiji Seika Kaisha Ltd., Japan
Effect of Myofibrillar Proteins of Sea Salmon (Pseudopercis semifasciata)- Malondehyde Interaction of Microstructure and Functionality. V.A. Tironi, M.C. Tomas, M.C. Anon, 1CIDCA, Argentina
Nitrates and Nitrites in Food. A. Telniceanu, Institute of Hygiene and Public Health Bucharest, Romania
Microstructure, Stability and Viscoelastic Properties of Reduced Oil Content Emulsions Formulated with Polysaccharides and Salt. J.M. Quintana1, A.N. Califano1, N.E. Zaritsky1, P. Partal2. 1 CIDCA, CONICET, Universidad Nacional de La Plata Argentina; 2 Universidad de Huelva, Escuela Politécnica Superior, Spain
A comparative study of connective tissue components in bovine and fish muscles using histological techniques. G. Enersen, R. Ofstad, V. Høst, K.H. Eggen, Norwegian Food Research Institute, Norway.
Effects on Texture and Microstructure after Replacing Manual by Mechanical Processing in Serra da Estrela Cheesemaking. P.J.M. Reis, F.X. Malcata. Escola Superior Biotecnologia - UCP, Portugal
Interfacial and Surface Behaviour of Protamine:BSA Mixed Systems. L.A. Glaser1, A.T. Paulson1, D. Rousseau2, 1Dalhousie University, Canada; 2Ryerson University, Canada
Soy protein isolate glycation and its effect on protein functionality. J. I. Boye1, P. Li2, J. Lenay3, V.A. Yaylayan2, F.K. Yeboah2, A. Achouri1; 1 Agriculture and Agri-Food Canada, Canada; 2 McGill University, Canada; 3University of Laval, Canada
The Role of Microstructure Changes in Microwave-Induced Toughness of Bread. M. Uzzan, E. Kesselman, O. Ramon, S. Mizrahi, I.J. Kopelman. Technion, I.I. T, Israel.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
At 06:23 AM 2/18/2002, you wrote: } Gary Gaugler wrote: } } } Why can't the student respond? She already did. } } Sorry I missed that. However, my intension is not just } to defend her.
Ah....then you have other intentions as well. OK. No problem.
} } Q1 and Q2 are so basic that I can't see how one } } would have to do much research to answer these } } questions. Reviewing Goldstein, Newbury, Echlin, } } et.al. should do the job.
The basic differentiating factor is the wavelength of light compared to that of electrons. As I "see" it. The shorter the wavelength used, the greater is the resolving power. Resolving power (resolution) is the ability to distinguish separation of two objects which are separated by a small distance. For LM, use RP=Lambda/2NA where Lambda is the wavelength of light, NA is the numerical aperture of the objective. For EM, use ThetaR=1.22Lambda/d. This angle, ThetaR, is the minimum angular separation between two objects based on wavelength and d, the diameter of the converging lens.
Thus, to increase resolution, either increase the diameter of the converging lens, use a shorter wavelength, or both. For light, 5500A is a good figure for its wavelength. What about the wavelength of electrons? One can take the particle view or the wave view. Since light is of the wave nature, we can similarly treat electrons. Huygens originally proposed the wave theory of light, in contrast to the particle theory of E=hv. De Broglie postulated that the wavelength of matter waves was the same as for light. This is given as Lambda=h/p which relates wavelength to momentum of the associated photons. Thus the duality of waves (Lambda and v) and that of particles (E and p). In some circumstances, light and electrons (matter) behave like a particle and in others, like a wave.
The wavelength of electrons can be computed using either the de Broglie relationship or the Bragg relationship. But if one uses the de Broglie relationship, which is based on momentum, then an interesting facet is exposed. That is, that the wavelength of electrons or an electron beam is dependent on its kinetic energy. Thus, the higher the KV of the beam, the shorter the wavelength and hence, better resolution. All other things being equal, of course.
Another advantage of the SEM is that because of its shorter wavelength of electrons, it supports a much greater working distance than does LM. But that's another story--if not a big difference between LM and SEM.
} Q3 might be a bit more involved. } } So you do think Q3 is not that simple.
} } I think that at least 99 out of 100 experts would } } answer Q1 and Q2 the same. At least, I would hope so. } } Let's assume the student receive two answers(since so many } of us refuse to answer). One of them happen to be is coming from } that 1 out 100. So to her, she will have two opposite answer } without knowing another 98 expert's opinion. What could she do?
She might actually have to do some on-line research to adjudicate the differences. Or, she may form her own hypothesis. Or she could give up.
} } Perhaps Raleigh should be contacted? } } That is exactly what I am planning to do! - if some one care to } post their answer. Will you? Please?
Sure. See above.
BTW, what did Raleigh have to say about this topic?
We have a SEM planchette containing 52 EDS standards that is in need of repolishing. It was originally made by C.M. Taylor Corporation of Stanford, CA but I am sure that other companies may be able to repolish it. I would appreciate receiving any information on this possibility.
Is it possible to do this ourselves? What would be involved?
Many thanks,
John B.
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Thank you so much for give the answer. I presume it is for Q2. How about Q1 and Q3?
Again, imagine I am the student, this is the conclusion I can get from your answer(after thinking, not just repeating you):
"If a light wave is about 500nm, I can expect the resolution about 250nm. Correct? Or maybe close?
} From your instruction on electron wave, I can calculate for a 10kv electron, the wavelength is about 0.013nm. Do I expect the resolution is about that? Maybe a little worse, 0.02nm? Or much worse, 0.2nm? Even at this resolution, I would be able to see single atom, right? I am very optimistic about this, even it is 100 times worse, I still have 2nm resolution. But there are much rooms to improve, if I wish to have 0.2nm resolution, all I have to do is to increase the energy by 100 times, so at 1000kv, I will have 0.2nm resolution and capable of seeing atoms, right?"
Could you please post the answer for Q1 and Q3, please? If no one else cares, at least I wish to learn.
I am completely surprised about the website you suggested on Q3. I though this is a "Question on electron microscope", not light microscope(see the subject title). If it is not restricted in electron microcopy, why should it be restricted in light microscopy? I am sure the term "temporal resolution" is used in many field.
Now can you see the point I am trying to make? These questions seems simple, but not all of us can answer it correctly, or at least without misleading(myself can be an example).
} } Gary Gaugler wrote: } } The basic differentiating factor is the wavelength of } light compared to that of electrons. As I "see" it. } The shorter the wavelength used, the greater is the } resolving power. Resolving power (resolution) is } the ability to distinguish separation of two objects } which are separated by a small distance. For LM, } use RP=Lambda/2NA where Lambda is the wavelength } of light, NA is the numerical aperture of the objective. } For EM, use ThetaR=1.22Lambda/d. This angle, } ThetaR, is the minimum angular separation between } two objects based on wavelength and d, the diameter } of the converging lens. } } Thus, to increase resolution, either increase the } diameter of the converging lens, use a shorter } wavelength, or both. For light, 5500A is a good } figure for its wavelength. What about the wavelength } of electrons? One can take the particle view or } the wave view. Since light is of the wave nature, } we can similarly treat electrons. Huygens originally } proposed the wave theory of light, in contrast to } the particle theory of E=hv. De Broglie postulated } that the wavelength of matter waves was the same } as for light. This is given as Lambda=h/p which } relates wavelength to momentum of the associated } photons. Thus the duality of waves (Lambda and v) } and that of particles (E and p). In some circumstances, } light and electrons (matter) behave like a particle } and in others, like a wave. } } The wavelength of electrons can be computed } using either the de Broglie relationship or the } Bragg relationship. But if one uses the de Broglie } relationship, which is based on momentum, } then an interesting facet is exposed. That is, that } the wavelength of electrons or an electron beam } is dependent on its kinetic energy. Thus, the } higher the KV of the beam, the shorter the wavelength } and hence, better resolution. All other things being } equal, of course. } } Another advantage of the SEM is that because of its } shorter wavelength of electrons, it supports a much } greater working distance than does LM. But that's } another story--if not a big difference between LM and SEM. } } } Q3 might be a bit more involved. } } } } So you do think Q3 is not that simple. } } Not for me. But here is some info: } } http://www.nasatech.com/Briefs/May01/NPO21056.html } } http://www.eun.org/kms/sites/eschola/view_leadingedge.cfm?oid=1714 } } http://www-celanphy.sci.kun.nl/Bruce%20web/scanning%20microscopy.htm
At 07:32 PM 2/18/2002, you wrote: } Dear Gary, } } Thank you so much for give the answer. } I presume it is for Q2. How about Q1 and Q3?
As far as I know, I answered all questions.... 1-3. At least, as best I could.
Were you able to contact Raleigh? That would be quite helpful.
} Again, imagine I am the student, this is the conclusion I can get } from your answer(after thinking, not just repeating you): } } "If a light wave is about 500nm, I can expect the resolution about } 250nm. } Correct? Or maybe close?
Not so, as I see it. 550nm (5500A) is the wavelength, but the resolution is dependent on the optics. How do you account for this? NA plays a major part of this facet. So does the condenser.
} } From your instruction on electron wave, I can calculate for a 10kv } electron, } the wavelength is about 0.013nm. Do I expect the resolution is about } that?
How do you calculate that? I do not deny that figure, I just would like to know how you arrived at that figure. Let me see the work.
} Maybe a little worse, 0.02nm? Or much worse, 0.2nm? Even at this } resolution, } I would be able to see single atom, right? } I am very optimistic about this, even it is 100 times worse, I still } have 2nm resolution. } But there are much rooms to improve, if I wish to have 0.2nm resolution, } all I have } to do is to increase the energy by 100 times, so at 1000kv, I will have } 0.2nm resolution } and capable of seeing atoms, right?"
Theoretically. But practically, who makes a SEM with 1MEV potential?? None that I know of. Even with TEM, 300KEV is quite high. But the difference of looking through or at a specimen is quite different...I think. It is morphology versus structure. Different.
But there is a more fundamental issue here. That is the distance of scan line dimensions. Say what?? Well, if the probe size is too large, it will overshadow the specimen. If it is too small, it will miss features of the specimen.
There are critical factors which affect the ultimate resolution of an EM. These are spacial aberration, chromatic aberration, and diffraction. But wait...there is more....diffraction limitations!! Aperture size. This affects intensity distribution and resolves on the disc of least confusion. Ah...but this brings us back to resolution. This is somewhat spot size (probe diameter) limited. If the spot size is too large, the scan will miss intermediate detail. If too small, it will also miss detail. The issue I think is the relationship of free working distance versus optical working distance. Both can be changed. Free WD is from the pole piece to the specimen while optical WD is the distance from the final aperture to the specimen. This is reflected in novel final lens designs.
} Could you please post the answer for Q1 and Q3, please? If no one else } cares, at least I wish to learn.
I believe that I did answer all three questions....in my own way.
} I am completely surprised about the website you suggested on Q3. I } though this } is a "Question on electron microscope", not light microscope(see the } subject title). } If it is not restricted in electron microcopy, why should it be } restricted in light } microscopy? I am sure the term "temporal resolution" is used in many } field.
Ah...therein lies the subtleties of the major issue. Unfortunately, it is not constrained to one venue. Were it not so, that would be terrific. Read and absorb the basic issue about temporal resolution. It seems to me that it is rather basic and intuitively obvious, once examined. Why would it make any difference between LM and EM, other than the inherent limitations therein?
} Now can you see the point I am trying to make? These questions seems } simple, but not all of us can answer it correctly, or at least without } misleading(myself can be an example).
Simple questions, per se, do not engender simple answers at all times. Such is the stuff of quantum mechanics and wave theory. But this is particularly different from Shrodinger, de Broglie and Bragg. Too many "negative waves," as was said in a notable video (Sutherland).
On Sat, 16 Feb 2002 xf200-at-attbi.com-at-sparc5.microscopy.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 } -----------------------------------------------------------------------. } } } It is not fair that the student does not have any chance to respond. } } If I were that student, here is my respond: } } You may think that giving me straight answer seems too easy on me. But } on my side, it is not that simple to use your answer. If I ask my } professor directly to get an answer, as long as I can repeat his/her } words like 3 years old, I can get a A, even if he is WRONG. } } But asking you is completely different. You could be WRONG too! If I } took your wrong answer straight to my professor, I will fail. No } matter how I explain this is from some "expert", it is not going to } help. } } Oh, how could YOU be wrong? Isn't that very insulting since you are } the expert? If you felt that way, I am sorry. I did not mean to insult } any one. However, how about post your answer to these "elementary" } questions on this list server? Do you think everyone on this list will } agree with you? } } Here are the re-post of Jenny Wang's original question: } 1. What are the advantages and disadvantages in using } electrons for microscopy rather than light? } 2. Does the wavelength of the electrons have anything } to do with the spatial resolution that the microscope } produces in the final picture? } 3. What is temporal resolution and how is it produced } in the electron microscope? } } Thank you very much in deed. } To coincide with Gary's comments I put forth my own:
Resolution in the transmission electron microscope (TEM) and the optical light microscope (OLM) are generally governed by the same laws of optical physics. Namely, the shorter the wavelength, the better the resolution. Thus, the advantage to the TEM would in fact be the ability to resolution smaller structures than the OLM.
The first issue is to define resolution. The second step is to define wavelength utilitzed, and the three step is to define numerical aperture. Once you have defined and understand these terms, only then will you realize how varied are your choices in drawing a final conclusion about your three questions.
An analogy would be to ask you, "Which is better, an airplane or a car", as it relates to transportation? Each have advantages and disadvantages.
I have written about wavelength and numerical aperture as they relate to resolution. However, you must keep in mind, as with transportation, there are a number of ways of getting around, so are there many caveats to adhering solely to these two perimeters.
Let's take for example Abbe's formula for OLM resolution. He states resolution is basically half the wavelength utilized. Consequently, if we use 500nm light, half would be 250nm resolution. The formula would be: resolution (d) = wavelength over 2 x numerical aperture (the n.a. of the condenser lens and the objective). If you take the numerical aperture of the condenser lens as 1.4 and the objective lens as 1.4 = 1.96 divided by the wavelength utilized (500nm) the resolution would be 255nm, or pretty close to Abbe's generalization of resolution equals half the wavelength utilitzed.
Now let's consider a 40x objective lens (n.a. 0.70) and the condenser lens (1.40). The resolution would be 510nm. Take a slide mounted with diatomaceous earth or perhaps a scrapping from the inside of your mouth and, using brightfield, tell me if you can resolve structures at 510nm? The issue is contrast! You cannot resolve the structures because these specimens are phase objects and not amplitude producing objects. The only way you may be able to spatially resolve any structure is to reduce the condenser aperture. You must reduce it to the point where in affect you reduce the annular aperture and thus, destroy the numerical aperture, one of the key components to Abbe's formula for resolution!
How do we increase the amplitude or contrast so as not to affect the n.a.? Well, darkfield illumination, phase contrast illumination, and differential interference contrast (DIC) illumination might be a start. The other method would be to stain the material, which would be difficult with diatomaceous earth!
These techniques are the hallmark for OLM, as they are methods of changing the amplitude through color as in staining the material, or refractive index as in darkfield, and phase/refractive indices as in phase contrast or DIC.
However, what happens when we want to observe structures smaller than 250nm? Yes, we can use fluorescence and gain slightly better resolution. However, we must resort to electrons...
The issue with the TEM and electron is not dissimilar to our issue with photons in the OLM. Spatial resolution is governed by sample thickness and accelerating voltage. If one uses an accelerating voltage of 100kV at a wavelight of 0.037, the resolving power is about 0.17nm.
However, let's decrease the kV to 50kV or place a thicker sample into electron beam. The consequence is an increase in scattered electrons, which equals less signal above background. Signal is good, background noise is bad. If we decrease our kV we get more contrast but less resolution, i.e., more amplitude through reduced resolution.
Spatial and temporal resolution are clearly more difficult to discuss in this space or my time. Again, sample thickness, accelerating voltage, and condenser/objective apertures (and even degree of vacuum) will play differing roles in the outcome of resolution. Then the question arises as to how you are going to document this resolution. It is visual, photographic, or digital/video?
Based on this information, do you have questions? Perhaps this information and that provided by others will help with the anwsers. However, who gets the grade for effort. Tell us (the members of the listserver) what you believe to be the correct answer(s). It is far easier to help through thoughtful questions as opposed to spewing out everything one knows on a given subject.
Ken ______________ Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203
John Bozolla wrote: ============================================================== We have a SEM planchette containing 52 EDS standards that is in need of repolishing. It was originally made by C.M. Taylor Corporation of Stanford, CA but I am sure that other companies may be able to repolish it. I would appreciate receiving any information on this possibility.
Is it possible to do this ourselves? What would be involved? ============================================================== SPI Supplies has been offering a standards "refurbishing" service for those with standards made by the C. M. Taylor Company (which by the way, does not exist any more). The prices are only slightly higher than those listed for the refurbishing of the SPI Standards, see URL http://www.2spi.com/catalog/standards/refurb.html
We have also acquired a large library of the original standards that were originally characterized by Dr. Taylor himself, and thererfore, if one or more of the individual standard items has fallen out, or comes out during the refurbishing, it can be replaced with essentially the identical standard
It is just over six months before ICEM-15 opens in Durban. Delegates are now registering and submitting abstracts, symposia chairs from throughout the world are busy organizing their symposia and awaiting the abstracts to see what has been submitted for their symposia, the invited speakers have been selected and are preparing their presentations, and in Durban, and elsewhere, the various members of the Organizing Committee and their helpers are doing whatever is necessary to make sure that their contribution towards this project is done properly and on time. Everything is in place, therefore, to make sure that delegates enjoy a most memorable and successful congress.
The Scientific Programme Committee, co-chaired by Mike Witcomb and Trevor Sewell, assisted by several key members of MSSA, the International Scientific Advisory Committee and the IFSEM executive, have put together what will be a wide-ranging and very interesting collection of topics into over 50 scientific symposia, four plenary sessions, the Ernst Abbe Lecture and the IFSEM Symposium. There will be at least 160 invited lectures by experts on these subjects, and because these speakers come from throughout the world, this will be a truly international meeting. The deadline for receipt of abstracts for the scientific symposia is 25 February.
The Technical Forum will provide an excellent opportunity for presentation and discussion of matters of a technical, commercial, managerial and administrative nature, in a pleasantly relaxed atmosphere that is conducive to fruitful discussion. This is something new for ICEM and it is hoped that it will be popular with many delegates especially managers, technicians, students and representatives of the trade. Requirements for abstracts of the Technical Forum are much less stringent than for the scientific symposia, and the deadline for receipt of these in Durban is 1 April, and thus allows delegates with “breaking news” quickly to present their results and get their abstracts in print!
The Trade Exhibition at ICEM-15 promises to provide an exciting array of instruments, accessories and consumables. At least 35 companies have reserved stands on the exhibition and a look at the exhibition plan on the website reveals that at this stage there are only 5 regular stands and a few display stands still available. The co-operation of the trade in making their arrangements so early is much appreciated.
The Local Organizing Committee and the Event Manager, Turners Conferences, are continuing with their impressive programme that is designed to ensure that all delegates, representatives, families and friends from wherever they may be leave Durban after ICEM-15 having happy memories of a most rewarding and enjoyable congress. Anyone doubting that Durban, or South Africa, has the ability to cater for events such as ICEM-15 can take heart from the success of two recent major international conferences that took place at ICC Durban, namely the world conferences on racism and HIV/AIDS. These both had registrations of 10 000 to 20 000 delegates and very few difficulties were experienced by delegates or the organizers. Further confidence has been expressed in South Africa’s record in handling international conferences through the award of the World Conference on Sustainable Development (Earth Summit 2002) that takes place in Johannesburg shortly before ICEM- 15. It has been reported that 194 heads of state will attend this meeting along with over 50 000 other delegates!
A special bonus for delegates is that due to recent movements in world currency markets the South African Rand has devalued significantly against most world currencies. This means that your dollars, euros, pesos, krones, etc, will all buy much more in the way of goods and services. Accommodation, even the very best, will appear to be ridiculously cheap by international standards, and not to be missed are the shopping opportunities, gourmet meals, wild life safaris, etc, that will represent amazing value for money before, during and after ICEM-15. BMW reported yesterday that its products are 25% cheaper in South Africa than anywhere else in the world so if you have room then now is the time to buy your new BMW! Accommodation is available to suit all tastes and budgets but prospective delegates are warned that the more popular hotels are filling up. Special rates have been negotiated with the official congress hotels, and these rates can be seen on the Turner’s web site (www.turners.co.za/icem15) and were included in the Second Circular and Call for Abstracts. Remember that today $US1.00 buys 11.50 SA Rands, 1 Euro buys 10 SA Rands and GBP1.00 buys 16.5 SA Rands.
South Africans are well known for their hospitality. Consequently the programme for social events has been well thought out and will provide something exciting, different and enjoyable for all ICEM-15 delegates, families and friends. Because of features for which South Africa is famous such as its cultural diversity, natural resources and great weather the social programme that goes with ICEM-15 will be an experience not to be missed. Details of all these activities are available on the web sites:
www.icem15.com www.turners.co.za/icem15
Getting to Durban, and around South Africa, is uncomplicated and surprisingly inexpensive when compared to Europe, North America and the East, especially if arrangements are made well in advance. As the “Earth Summit” is being held in Johannesburg shortly before ICEM-15 it is all the more important for ICEM- 15 delegates to make their travel arrangements well in advance. There are many flights daily by most of the world’s major airlines from Europe, North and South America, the East and Australasia to Durban, either directly or via Johannesburg or Cape Town. Travel networks within South Africa, whether air, road or rail, are well developed and by air most major destinations can be reached from Durban within two hours, making use of up-to-date fleets of aircraft used by South African Airways and other domestic airlines. Airport formalities and transfers are straightforward, and to give delegates an early welcome (and be there for anyone requiring advice or assistance) the ICEM Organizing Committee will have welcome desks in the arrivals halls of Johannesburg, Cape Town and Durban International Airports.
The Event Managers for ICEM-15, Turners Conferences and Conventions, will be pleased to answer any enquiries about flights, tours, accommodation, car hire, etc, and have negotiated the most favourable rates for ICEM-15 delegates. Delegates who have already made their bookings through Turners are very pleased with the competitive fares that they have been offered. For more information contact Mr Dudley Randall (Turners Conferences) by email at turner17-at-galileosa.co.za
Look also the following web sites for more relevant information: http://www.icem15.com (official ICEM-15 web site) http://www.turners.co.za/icem15 (event management web site) http://www.uct.ac.za/depts/emu/mssa (MSSA web site) http://www.materials.ox.ac.uk/ifsem (IFSEM web site) http://www.satour.com (SA Tourism web site) http://southafrica.net (tourist information web site) http://www.kwazulu-natal.co.za (Durban and surrounding area tourist information)
The ICEM-15 Organizing Committee looks forward to welcoming you to Durban in September 2002. It will be an experience not to be missed.
Dorothy, I have looked at beta-gal in the EM (Cohen-Gould & Mikawa, Developmental Biol 177(1) 265-276, 1996). the X-gal crystals are very easy to see in the EM. The main caveat, is that the embedding resing doesn't penetrate the crystals, so that your sections tend to tear under the beam. I used parlodion or formvar coated grids to add support and that helped a lot. Good luck, Lee -- Lee Cohen-Gould Electron & Optical Microscopy Facilities Weill Medical College of Cornell U. (212)746-6146 Rms A-105, LC-207
We looking for an EDS system in good working condition with horizontal entry detector (Be window), electronics console, monitor, and printer. We prefer a system that can be directly mounted on a Philips TEM but other systems could work.
The large variation in hardness between the different standards is probablythe biggest obstacle in attempting this yourself. If another standards supplier will guarantee results you should strongly consider that. Otherwise, if you have someone withexperience in critical metallurgical preparation, who also has experiencewith nonmetallics such as ceramics, you might enlist their help. It would not be wise to learn on your standards.
The nature of the problem to be corrected will determine the steps to take. You should use the minimum polish size possible in order to avoid creating new problems. If your intention is merely to remove beam contamination, 1 micron alumina should be more than adequate and you will need to follow that with at least one step of finer polishing. If the standards have been damaged by excessive beam currents so that the existing surface needs to be removed, you will need to deal with the problem of relief development among materials of differing hardness.
In general, to avoid this it is best to use an abrasive of high efficiency on a napless cloth. 3 micron diamond on woven nylon would be an example. The time and pressure should be kept to a minimum in order to avoid rounding of edges and embedding of abrasive in softer standards, respectively. You will have to follow this with exhaustive cleaning followed by polishing steps. This regimen has worked well for me with composite assemblies involving gold (very malleable), soft solder (malleable but with some brittle intermetallic compounds), plating layers (which must not be deformed), steel (prone to rapid oxidation), and 96% alumina/ 4% silica ceramic (hardness nearly Mhos 9) in the same sample.
The final issue is what polishing fluid to use. Years ago it was alleged that some of the standards suppliers used olive oil, or a mixture containing that, rather than something more "high tech". Olive oil is mostly oleic acid, so one would assume that it has some detergent properties as well as the capability to form fatty acid soaps with a wide variety of metals, including most copper alloys. So while it may offer some advantages, it is not inert, and should be used intelligently.
Removal of the last traces of very fine polish may be difficult from some surfaces. In my experience a detergent that functions in a non-aqueous medium is very helpful if used (briefly!) with ultrasonic agitation. Potassium methyl cyclohexyl oleate (Vulpex) in naphtha is compatible with most standards materials but you should expect to wash repeatedly in clean solvent and dry under vacuum.
You should take special precautions with certain standards. For example, some sulfides, arsenides and selenides are susceptible to polar organic solvents that might come to mind as rinse agents.
I would be interested, as well, in hearing from anyone on the list who has successfully reworked their standards or provides this service.
John Twilley Conservation Scientist
John J. Bozzola 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 } -----------------------------------------------------------------------. } } } We have a SEM planchette containing 52 EDS standards that is in need } of repolishing. It was originally made by C.M. Taylor Corporation of } Stanford, CA but I am sure that other companies may be able to } repolish it. I would appreciate receiving any information on this } possibility. } } Is it possible to do this ourselves? What would be involved? } } Many thanks, } } John B. } }
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Sorry I must have missed something. If you are the student and give the professor some website as an answer, I doubt you will pass it. I know I won't.. So anybody else can post the answer on Q3 - in some simple sentence?
You did say some advantage of EM over LM, but I was expecting "better resolution", "expensiveness" which student can directly relate to. I don't think they know what "working distance" means if they have not used SEM before.
To figure out wavelength is not that difficult, just use the formula you gave me. lambda = h/p (h is plank constant) p is momentum, = mv v is the speed, as in E=mv^2/2, E is energy, = eV, V is potential(voltage). So knowing V, we can get lambda. Of course you have to include relativistic effect in electron mass m = m0/sqrt(1-(v/c)^2). (By the way, this is a "fundamental" question too, but I am very happy to answer it, it at least helps me refreshing my memory)
I do not think we have much disagreement on the resolution of LM. The textbook says the resolution is about 200nm. Sure you can get worse, but that is still in the same order of the wavelength.
It is greatly different in EM. It is hundreds of times worse. That's fine considering the optics. But at least it should be proportional to wavelength, i.e. "shorter wavelength, better resolution". right? This is NOT the case in SEM. It is not impossible to build a 1MeV SEM. No one did that. Not even 100kv. Why? Simply because it does not improve resolution! To shorten the wavelength, the only way to do is to increase energy (see the relation above).Increase energy will definitely decrease the practical resolution. It might improve the spherical abberation. The overall effect is that : shortening wavelength does not necessarily improve resolution in SEM. Theoratically.
I could not find any definition of "temporal resolution" in electron microscope textbook. So I thought "temporal resolution" might relates to "temporal coherency" which effects resolution. We believe that we need a better temporal coherent source to produce better resolution. e.g. use very narrow energy distribution(through a very stable high voltage). However Lord Raleigh says: "Incoherent source produce BETTER resolution than coherent source".
I found discussing these fundamental questions fascinating.
Gary Gaugler wrote:
} At 07:32 PM 2/18/2002, you wrote: } } Dear Gary, } } } } Thank you so much for give the answer. } } I presume it is for Q2. How about Q1 and Q3? } } As far as I know, I answered all questions.... } 1-3. At least, as best I could. } } Were you able to contact Raleigh? That would } be quite helpful. } } } Again, imagine I am the student, this is the conclusion I can get } } from your answer(after thinking, not just repeating you): } } } } "If a light wave is about 500nm, I can expect the resolution about } } 250nm. } } Correct? Or maybe close? } } Not so, as I see it. 550nm (5500A) is the wavelength, } but the resolution is dependent on the optics. How do you } account for this? NA plays a major part of this facet. So } does the condenser. } } } } From your instruction on electron wave, I can calculate for a 10kv } } electron, } } the wavelength is about 0.013nm. Do I expect the resolution is about } } that? } } How do you calculate that? I do not deny that figure, I just } would like to know how you arrived at that figure. Let me } see the work. } } } Maybe a little worse, 0.02nm? Or much worse, 0.2nm? Even at this } } resolution, } } I would be able to see single atom, right? } } I am very optimistic about this, even it is 100 times worse, I still } } have 2nm resolution. } } But there are much rooms to improve, if I wish to have 0.2nm resolution, } } all I have } } to do is to increase the energy by 100 times, so at 1000kv, I will have } } 0.2nm resolution } } and capable of seeing atoms, right?" } } Theoretically. But practically, who makes a SEM with 1MEV } potential?? None that I know of. Even with TEM, 300KEV is } quite high. But the difference of looking through or at a } specimen is quite different...I think. It is morphology versus } structure. Different. } } But there is a more fundamental issue here. That is } the distance of scan line dimensions. Say what?? } Well, if the probe size is too large, it will overshadow } the specimen. If it is too small, it will miss features } of the specimen. } } There are critical factors which affect the ultimate } resolution of an EM. These are spacial aberration, } chromatic aberration, and diffraction. But wait...there } is more....diffraction limitations!! Aperture size. This } affects intensity distribution and resolves on the } disc of least confusion. Ah...but this brings us back to } resolution. This is somewhat spot size (probe diameter) } limited. If the spot size is too large, the scan will miss } intermediate detail. If too small, it will also miss detail. } The issue I think is the relationship of free working } distance versus optical working distance. Both can be } changed. Free WD is from the pole piece to the } specimen while optical WD is the distance from the } final aperture to the specimen. This is reflected in } novel final lens designs. } } } Could you please post the answer for Q1 and Q3, please? If no one else } } cares, at least I wish to learn. } } I believe that I did answer all three questions....in my own way. } } } I am completely surprised about the website you suggested on Q3. I } } though this } } is a "Question on electron microscope", not light microscope(see the } } subject title). } } If it is not restricted in electron microcopy, why should it be } } restricted in light } } microscopy? I am sure the term "temporal resolution" is used in many } } field. } } Ah...therein lies the subtleties of the major issue. Unfortunately, } it is not constrained to one venue. Were it not so, that would be } terrific. Read and absorb the basic issue about temporal } resolution. It seems to me that it is rather basic and } intuitively obvious, once examined. Why would it make any } difference between LM and EM, other than the inherent limitations } therein? } } } Now can you see the point I am trying to make? These questions seems } } simple, but not all of us can answer it correctly, or at least without } } misleading(myself can be an example). } } Simple questions, per se, do not engender simple answers } at all times. Such is the stuff of quantum mechanics and } wave theory. But this is particularly different from Shrodinger, } de Broglie and Bragg. Too many "negative waves," as was } said in a notable video (Sutherland). } } Gary Gaugler, Ph.D.
I suppose it is good to go back and rehash some of things on a more fundamental level. Even those of us who have been doing this for many years get rusty and can use a refresher course.
It sounds like you are touching on another factor that practically determines resolution, and that is the interaction volume - the scattering of the beam once it encounters the specimen. I often tell students who ask me about the probe diameter of the SEM that they are probably asking the wrong question, especially when dealing with x-ray analyses. At probe currents of less than 1 nA, the diameter of the probe is quite a bit smaller than the x-ray interaction volume. Under those conditions, the accelerating voltage is more critical in determining the chemical spatial resolution than is the beam diameter. The student will need to be aware of these other factors in addition to the wavelength limit of the resolution.
"Temporal resolution" is an ambiguous term to me, too. I replied to the student asking if she was referring to the speed with which images could be collected in order to observe changes in the sample. (Someone was definitely asking about observing processes in cells. It may not have been her.) I did not get a reply, but if my guess is correct, we then need to be talking about signal-to-noise ratios. I can take high-resolution SEM images, but it normally requires sampling over a fairly slow scan. TV rate images can be obtained, but the noise becomes a very significant factor. It seems to me that most (not all) LM images have much better S/N ratios.
The issue of documenting biological processes raised a whole other issue. Most biological processes stop long before the sample is introduced into the EM, except maybe for some special cases in an E-SEM. Certainly, the intra-cellular processes are not visible in the EM.
I can see some worth to this whole exercise if the instructor is trying to get the students to think through the strengths and weaknesses of EM vs. LM and also SEM vs. TEM. However, it is always "interesting" to see these questions come across the list. The intent of the exercise is not always clear, and the questions are not always formulated well. But I guess it provides some relief from the routine.
Warren
At 01:37 PM 2/19/02 -0500, you wrote: } Gary, } } Sorry I must have missed something. If you are the student and give the } professor some website as an answer, I doubt you will pass it. I know I } won't.. } So anybody else can post the answer on Q3 - in some simple sentence? } } You did say some advantage of EM over LM, but I was expecting } "better resolution", "expensiveness" which student can directly } relate to. I don't think they know what "working distance" means } if they have not used SEM before. } } To figure out wavelength is not that difficult, just use the formula you } gave me. lambda = h/p (h is plank constant) p is momentum, = mv } v is the speed, as in E=mv^2/2, E is energy, = eV, V is potential(voltage). } So knowing V, we can get lambda. Of course you have to include } relativistic effect in electron mass m = m0/sqrt(1-(v/c)^2). } (By the way, this is a "fundamental" question too, but I am very happy } to answer it, it at least helps me refreshing my memory) } } I do not think we have much disagreement on the resolution of LM. } The textbook says the resolution is about 200nm. Sure you can get } worse, but that is still in the same order of the wavelength. } } It is greatly different in EM. It is hundreds of times worse. That's fine } considering the optics. But at least it should be proportional to } wavelength, } i.e. "shorter wavelength, better resolution". right? } This is NOT the case in SEM. It is not impossible to build a 1MeV SEM. } No one did that. Not even 100kv. Why? Simply because it does not improve } resolution! To shorten the wavelength, the only way to do is to increase } energy } (see the relation above).Increase energy will definitely decrease the } practical resolution. It might improve the spherical abberation. The } overall effect is that : shortening wavelength does not necessarily } improve resolution in SEM. Theoratically. } } I could not find any definition of "temporal resolution" } in electron microscope textbook. So I thought "temporal resolution" } might relates to "temporal coherency" which effects resolution. } We believe that we need a better temporal coherent source to produce } better resolution. e.g. use very narrow energy distribution(through a very } stable } high voltage). However Lord Raleigh says: } "Incoherent source produce BETTER resolution than coherent source". } } I found discussing these fundamental questions fascinating.
Thank you so much for your answer. I learned a lot. My intention is not to show that I have better answer than other people. In fact, it is quite the opposite. The intention is mostly it is selfish - I wish to learn myself.
Anyway, if you wish to know my answer, here it is: 1. What are the advantages and disadvantages in using electrons for microscopy rather than light? Answer: Advantage: Better resolution, Better depth of field/focus, etc Disadvatage: Expansive, more complicated sample preparation, etc. However, I think these answers are misleading. Someone might think if she/he have enough money, great effort, she/he can accomplish the tasks with light microscope with better resolution simple by switching to electron microscope.
If I ask all the light microscopist, how many of your tasks can be simply replaced with electron microscopy - presume you have enough funding and man power?
If the professor wish to get the answer above, it is better re-phrased as: "a. List the advantages and disadvantages of electron microscopy. b. List the advantages and disadvantages of light microscopy?"
Like the analogy you ask , "Which is better, an airplane or a car", as it relates to transportation? Each have advantages and disadvantages. This is a perfect general question. You can compare them. However, if you ask "What is the advantage of an airplan rather than car if I wish to travel from A to B?" You might say "airplan is faster". Now if you are late to work in the morning, would airplan help you?
I might be fussy. But to avoid misleading, any effort is worthwhile.
2. Does the wavelength of the electrons have anything to do with the spatial resolution that the microscope produces in the final picture? Answer: Generally, the shorter the wavelength, the better the resolution. This is particularly true in light microscope. The resolving power is about the same magnitude as wavelength, more or less. However, in electron microscope, there is some limit. Even everything else is unchanged, e.g. optics and sample. Shortening wavelength does not garantee increase resolution. For example, in SEM, shorter wavelength means higher voltage(no other way), which means increasing interaction volumn, therefore it is also a factor of reducing resolution. A balance can be made at certain voltage, in another word, wavelength. So there is an optimal wavelength to produce best resolution in SEM.
By the way, in your following comments, at 100kv, the wavelength is 0.0037nm (or 0.037A), compare to 0.17nm resolution. There is a huge difference between
light and electron microscope resolution relative to their wavelength.
3. What is temporal resolution and how is it produced in the electron microscope?
Honestly, I don't know. I know what "temporal resolution" of human eye is which is the base for motion picture. There are definitions in light microscopy. But I could not find anything in simple EM textbook. There is a website given by Gary at: http://www.nasatech.com/Briefs/May01/NPO21056.html which tells the "The principle of stroboscopy would be extended to scanning electron microscopy" where SEM have adquate temporal resolution.
Yours sincerely, Xudong Fan fanx-at-msu.edu
Ken Tiekotter wrote:
} To coincide with Gary's comments I put forth my own: } } Resolution in the transmission electron microscope (TEM) and the optical } light microscope (OLM) are generally governed by the same laws of optical } physics. Namely, the shorter the wavelength, the better the resolution. } Thus, the advantage to the TEM would in fact be the ability to resolution } smaller structures than the OLM. } } The first issue is to define resolution. The second step is to define } wavelength utilitzed, and the three step is to define numerical aperture. } Once you have defined and understand these terms, only then will you } realize how varied are your choices in drawing a final conclusion about } your three questions. } } An analogy would be to ask you, "Which is better, an airplane or a car", } as it relates to transportation? Each have advantages and disadvantages. } } I have written about wavelength and numerical aperture as they relate to } resolution. However, you must keep in mind, as with transportation, there } are a number of ways of getting around, so are there many caveats to } adhering solely to these two perimeters. } } Let's take for example Abbe's formula for OLM resolution. He states } resolution is basically half the wavelength utilized. Consequently, if we } use 500nm light, half would be 250nm resolution. The formula would be: } resolution (d) = wavelength over 2 x numerical aperture (the n.a. of the } condenser lens and the objective). If you take the numerical aperture of } the condenser lens as 1.4 and the objective lens as 1.4 = 1.96 divided by } the wavelength utilized (500nm) the resolution would be 255nm, or pretty } close to Abbe's generalization of resolution equals half the wavelength } utilitzed. } } Now let's consider a 40x objective lens (n.a. 0.70) and the condenser lens } (1.40). The resolution would be 510nm. Take a slide mounted with } diatomaceous earth or perhaps a scrapping from the inside of your mouth } and, using brightfield, tell me if you can resolve structures at 510nm? } The issue is contrast! You cannot resolve the structures because these } specimens are phase objects and not amplitude producing objects. The only } way you may be able to spatially resolve any structure is to reduce the } condenser aperture. You must reduce it to the point where in affect you } reduce the annular aperture and thus, destroy the numerical aperture, one } of the key components to Abbe's formula for resolution! } } How do we increase the amplitude or contrast so as not to affect the n.a.? } Well, darkfield illumination, phase contrast illumination, and } differential interference contrast (DIC) illumination might be a start. } The other method would be to stain the material, which would be difficult } with diatomaceous earth! } } These techniques are the hallmark for OLM, as they are methods of changing } the amplitude through color as in staining the material, or refractive } index as in darkfield, and phase/refractive indices as in phase contrast } or DIC. } } However, what happens when we want to observe structures smaller than } 250nm? Yes, we can use fluorescence and gain slightly better resolution. } However, we must resort to electrons... } } The issue with the TEM and electron is not dissimilar to our issue with } photons in the OLM. Spatial resolution is governed by sample thickness } and accelerating voltage. If one uses an accelerating voltage of 100kV at } a wavelight of 0.037, the resolving power is about 0.17nm. } } However, let's decrease the kV to 50kV or place a thicker sample into } electron beam. The consequence is an increase in scattered electrons, } which equals less signal above background. Signal is good, background } noise is bad. If we decrease our kV we get more contrast but less } resolution, i.e., more amplitude through reduced resolution. } } Spatial and temporal resolution are clearly more difficult to discuss in } this space or my time. Again, sample thickness, accelerating voltage, and } condenser/objective apertures (and even degree of vacuum) will play } differing roles in the outcome of resolution. Then the question arises as } to how you are going to document this resolution. It is visual, } photographic, or digital/video? } } Based on this information, do you have questions? Perhaps this } information and that provided by others will help with the anwsers. } However, who gets the grade for effort. Tell us (the members of the } listserver) what you believe to be the correct answer(s). It is far } easier to help through thoughtful questions as opposed to spewing out } everything one knows on a given subject. } } Ken } ______________ } Ken Tiekotter, Adjunct Professor } The University of Portland } Department of Biology } 5000 N. Willamette Blvd. } Portland, OR 97203
At 10:37 AM 2/19/2002, you wrote: } Gary, } } Sorry I must have missed something. If you are the student and give the } professor some website as an answer, I doubt you will pass it. I know I } won't.. } So anybody else can post the answer on Q3 - in some simple sentence?
The URLs are not the answer. What is at the URLs is. You have to read the material at the URLs to compose an answer for yourself. Better still, look up the dictionary definition of "temporal" and then read the material at the URLs. If you still don't get it, let me know.
} You did say some advantage of EM over LM, but I was expecting } "better resolution", "expensiveness" which student can directly } relate to. I don't think they know what "working distance" means } if they have not used SEM before.
I thought I did say better resolution and also working distance for SEM. Working distance applies to LM and EM. At upper magnifications, the LM user is well aware of this factor. I am primarily talking about SEM specifically rather than TEM when discussing EM. I'm not a TEM person. I do SEM and LM.
} To figure out wavelength is not that difficult, just use the formula you } gave me. lambda = h/p (h is plank constant) p is momentum, = mv } v is the speed, as in E=mv^2/2, E is energy, = eV, V is potential(voltage). } So knowing V, we can get lambda. Of course you have to include } relativistic effect in electron mass m = m0/sqrt(1-(v/c)^2). } (By the way, this is a "fundamental" question too, but I am very happy } to answer it, it at least helps me refreshing my memory) } } I do not think we have much disagreement on the resolution of LM. } The textbook says the resolution is about 200nm. Sure you can get } worse, but that is still in the same order of the wavelength.
There are new LM instruments out there, like confocal and scanning confocal, which have excellent resolution for LM. But SEM still beats them.
} It is greatly different in EM. It is hundreds of times worse. That's fine } considering the optics. But at least it should be proportional to } wavelength, } i.e. "shorter wavelength, better resolution". right? } This is NOT the case in SEM. It is not impossible to build a 1MeV SEM. } No one did that. Not even 100kv. Why? Simply because it does not improve } resolution! To shorten the wavelength, the only way to do is to increase } energy } (see the relation above).Increase energy will definitely decrease the } practical resolution. It might improve the spherical abberation. The } overall effect is that : shortening wavelength does not necessarily } improve resolution in SEM. Theoratically.
Let's see. In SEM, resolution is inversely proportional to wavelength of the electrons. Shorter wavelength, greater resolution, in general. Wavelength of electrons is inversely proportional to energy. Higher energy, shorter wavelength. So, higher voltage/energy, higher resolution (more resolving power). My old SX-40 brochure says that it had a resolution of 60A. Newer SEMs are better than that. My SEM is newer than the SX-40 and is supposed to have a resolution of about 40A. I guess so, but I can't measure it. I tried but failed. I get about 120-180A at 15KV. And that's an eyeball guess. I cannot find a perfectly sharp edge!
BTW, I've been told that 1MV TEMs do exist. Most SEMs are up to 30KV.
} I could not find any definition of "temporal resolution" } in electron microscope textbook. So I thought "temporal resolution" } might relates to "temporal coherency" which effects resolution. } We believe that we need a better temporal coherent source to produce } better resolution. e.g. use very narrow energy distribution(through a very } stable } high voltage). However Lord Raleigh says: } "Incoherent source produce BETTER resolution than coherent source".
That is a coherent statement but wrong temporal at this time.
} I found discussing these fundamental questions fascinating.
I've been thinking about this problem and have a theoretical solution based on unknown hardware.
If the CCTV of the SX-40 is indeed RS-170, then there is hope. The key is, it seems to me, is to perform Kalman filter operations on each field and frame until the completed image is good enough to grab.
To do this would require a frame grabber board with a controllable buffer or a software product that would malloc() memory in the PC for one or more buffers. Thus, the issue whether the final image is grabbed from the frame grabber board or from PC memory via the interface/control software. But either way, it would yield a high quality 640x480 pixel image. This is a perfectly acceptable size for most work. If higher rez is needed, then the expensive stuff comes into play.
gary g.
At 10:27 AM 2/6/2002, you wrote:
} Dear Lister } } I would like to thank those who gave me some input to my first } delimma. } } I found that there are a ton of different frame grabbers out } there. What I dont know is what kind of output signal the scope produces, as } far as my understanding of the ISI SX-40 SEM it puts a signal out to the } CRT. Some of the venders' questions have been if it is an NTSC, PAL, or } RS170 type signal.I am slightly familiar with the first two, but the latter I } have no idea. I do know that there are a couple of "off the shelf" products } from GW Electronics and Image Slave, but these setups are priced at } $4000+. So what I am looking for is if someone } can point me in the direction of where I can reseach this information.
} Your message cannot be delivered to the following recipients: } } Recipient address: wil1323-at-IMAP2.ASU.EDU } Original address: curari-at-asu.edu } Reason: Remote SMTP server has rejected address } Diagnostic code: smtp;550 5.1.1 {wil1323-at-IMAP2.ASU.EDU} ... User unknown } Remote system: dns;imap2.asu.edu } (TCP|129.219.110.72|44165|129.219.110.75|7025)
Shorter wavelength=greater resolution may be the general rule for all imaging systems using waves, but in SEMs that is far from the whole story, and no commercial TEM comes anywhere close to the theoretical resolution limit. TEM resolving power is spoiled by lens aberrations. I remember a recent paper which described a way of neutralising spherical aberrations in a TEM using an electron mirror.
Early SEM guns and optics were designed for 30 kV, and were virtually unusable at 1kV. SEM development since the 60s has been about emitters, guns and optics. Now, with FEGSEMs we can see that although the fundamental rules still apply, major improvements in overall performance at low kV have been possible by using near-monochromatic emitters. Resolution in uncoated light-element specimens is now seen to be a trade-off between kV/wavelength and beam interaction volume. In the surface of a potato starch grain, starch crystal edges can be seen at 1 or 2kV, but these are obliterated at 5 or 10 kV as the beam interaction volume grows. Resolution in TEM is also a function of contrast, which is poorer at high kV, which is why some biological TEMs are designed with long working distance optics, trading some resolving power to buy higher contrast. EM instruments have greater theoretical RESOLVING POWER at higher kV, but in practice the RESOLUTION can be poorer. Resolving power is an instrument property. Resolution can be a specimen or image property. The two do not always coincide.
Chris
} Let's see. In SEM, resolution is inversely proportional to wavelength } of the electrons. Shorter wavelength, greater resolution, in general. } Wavelength of electrons is inversely proportional to energy. Higher } energy, shorter wavelength. So, higher voltage/energy, higher } resolution (more resolving power). My old SX-40 brochure says } that it had a resolution of 60A. Newer SEMs are better than that. } My SEM is newer than the SX-40 and is supposed to have a } resolution of about 40A. I guess so, but I can't measure it. } I tried but failed. I get about 120-180A at 15KV. And that's } an eyeball guess. I cannot find a perfectly sharp edge!
} Dear Ken, } } Thank you so much for your answer. I learned a lot. } My intention is not to show that I have better answer than other people. } In fact, it is quite the opposite. The intention is mostly it is } selfish - I wish to learn myself. } } Anyway, if you wish to know my answer, here it is: } 1. What are the advantages and disadvantages in using } electrons for microscopy rather than light? } Answer: Advantage: } Better resolution, Better depth of field/focus, etc } Disadvatage: } Expansive, more complicated sample preparation, etc. } However, I think these answers are misleading. Someone might think if she/he } have enough money, great effort, she/he can accomplish the tasks with light } microscope with better resolution simple by switching to electron microscope.
*You must specify TEM or SEM or STEM or ESEM. You cannot ask these questions as to how they relate solely to "EM", as the answers relating to physical design and the physics associated with their operation are unique to each instrument.
You state "better depth of field/focus". Yes, in the SEM the scanning probe instruments, depth of field/focus is important. However, depth of field in a 90nm section for TEM is, as you can imagine, extremely small, hence the advantage of accelerating voltage.
As for the disadvantages of EM as being labor intensive, have you any knowledge of LM preparation or the difference between TEM and SEM preparation? SEM preparation is a walk in the park compared to the rigors necessary to preserve, examine, and document ultrastrutural resolution at 0.1nm.
As for throwing money at the issue, the issue is clear. Think of your compliment of microscopes as a toolbox: each tool in the toolbox has a specific function. Yes, you can drive a nail in the wall to hung a picture with the handle of a screwdriver, however, the best tool would be a hammer!
The light microscope does not have the resolution of a TEM or the depth of focus of a SEM, however, can a SEM or TEM convert phase differences of amplitude as the phase contrast OLM? Can the SEM or TEM differentiate specific structures using histological stain producing color difference or look at refractive index differences as in DIC: I think not.
However, the SEM can provide incredible information about surface topography, provide elemental differention, and 500x the depth of focus of an OLM. Can it image surface information: NO, and not a any kV or spot size. And then there is the TEM, STEM, ESEM....
As for instrument expense, look into a multi-photon, scanning laser confocal OLM microscope at $400k to $600K!
} If I ask all the light microscopist, how many of your tasks can be } simply replaced with electron microscopy - presume you have enough } funding and man power?
Toolbox...more expensive toys do not necessarily answer the simplest questions
} } If the professor wish to get the answer above, it is better re-phrased as: } "a. List the advantages and disadvantages of electron microscopy. } b. List the advantages and disadvantages of light microscopy?" } } Like the analogy you ask , "Which is better, an airplane or a car", } as it relates to transportation? Each have advantages and disadvantages. } This is a perfect general question. You can compare them. } However, if you ask "What is the advantage of an airplan rather than } car if I wish to travel from A to B?" You might say "airplan is faster". } Now if you are late to work in the morning, would airplan help you? } } I might be fussy. But to avoid misleading, any effort is worthwhile. } } 2. Does the wavelength of the electrons have anything } to do with the spatial resolution that the microscope } produces in the final picture? } Answer: Generally, the shorter the wavelength, the better the resolution. } This is particularly true in light microscope. The resolving power is about } the same magnitude as wavelength, more or less.
Do you not agree there is a limit to the resolution of the OLM regardless of wavelength or with respect to wavelength? If the OLM resolution were limitless with respect to wavelength, why bother with the TEM?
} However, in electron microscope, there is some limit. Even everything } else is unchanged, e.g. optics and sample. Shortening wavelength does } not garantee increase resolution. For example, in SEM, shorter } wavelength means higher voltage(no other way), which means increasing } interaction volumn, therefore it is also a factor of reducing } resolution. A balance can be made at certain voltage, in another word, } wavelength. So there is an optimal wavelength to produce best } resolution in SEM.
In the TEM higher voltage IS resolution, or at least a key component. } } By the way, in your following comments, at 100kv, the wavelength is 0.0037nm } (or 0.037A), compare to 0.17nm resolution. There is a huge difference between } light and electron microscope resolution relative to their wavelength. } } 3. What is temporal resolution and how is it produced } in the electron microscope? } } Honestly, I don't know. I know what "temporal resolution" of human eye } is which is the base for motion picture. There are definitions in } light microscopy. But I could not find anything in simple EM textbook. } There is a website given by Gary at: } http://www.nasatech.com/Briefs/May01/NPO21056.html which tells the } "The principle of stroboscopy would be extended to scanning electron } microscopy" where SEM have adquate temporal resolution. } } Yours sincerely, } Xudong Fan } fanx-at-msu.edu } Enough is enough... kt -- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203
} Ken Tiekotter wrote: } } } To coincide with Gary's comments I put forth my own: } } } } Resolution in the transmission electron microscope (TEM) and the optical } } light microscope (OLM) are generally governed by the same laws of optical } } physics. Namely, the shorter the wavelength, the better the resolution. } } Thus, the advantage to the TEM would in fact be the ability to resolution } } smaller structures than the OLM. } } } } The first issue is to define resolution. The second step is to define } } wavelength utilitzed, and the three step is to define numerical aperture. } } Once you have defined and understand these terms, only then will you } } realize how varied are your choices in drawing a final conclusion about } } your three questions. } } } } An analogy would be to ask you, "Which is better, an airplane or a car", } } as it relates to transportation? Each have advantages and disadvantages. } } } } I have written about wavelength and numerical aperture as they relate to } } resolution. However, you must keep in mind, as with transportation, there } } are a number of ways of getting around, so are there many caveats to } } adhering solely to these two perimeters. } } } } Let's take for example Abbe's formula for OLM resolution. He states } } resolution is basically half the wavelength utilized. Consequently, if we } } use 500nm light, half would be 250nm resolution. The formula would be: } } resolution (d) = wavelength over 2 x numerical aperture (the n.a. of the } } condenser lens and the objective). If you take the numerical aperture of } } the condenser lens as 1.4 and the objective lens as 1.4 = 1.96 divided by } } the wavelength utilized (500nm) the resolution would be 255nm, or pretty } } close to Abbe's generalization of resolution equals half the wavelength } } utilitzed. } } } } Now let's consider a 40x objective lens (n.a. 0.70) and the condenser lens } } (1.40). The resolution would be 510nm. Take a slide mounted with } } diatomaceous earth or perhaps a scrapping from the inside of your mouth } } and, using brightfield, tell me if you can resolve structures at 510nm? } } The issue is contrast! You cannot resolve the structures because these } } specimens are phase objects and not amplitude producing objects. The only } } way you may be able to spatially resolve any structure is to reduce the } } condenser aperture. You must reduce it to the point where in affect you } } reduce the annular aperture and thus, destroy the numerical aperture, one } } of the key components to Abbe's formula for resolution! } } } } How do we increase the amplitude or contrast so as not to affect the n.a.? } } Well, darkfield illumination, phase contrast illumination, and } } differential interference contrast (DIC) illumination might be a start. } } The other method would be to stain the material, which would be difficult } } with diatomaceous earth! } } } } These techniques are the hallmark for OLM, as they are methods of changing } } the amplitude through color as in staining the material, or refractive } } index as in darkfield, and phase/refractive indices as in phase contrast } } or DIC. } } } } However, what happens when we want to observe structures smaller than } } 250nm? Yes, we can use fluorescence and gain slightly better resolution. } } However, we must resort to electrons... } } } } The issue with the TEM and electron is not dissimilar to our issue with } } photons in the OLM. Spatial resolution is governed by sample thickness } } and accelerating voltage. If one uses an accelerating voltage of 100kV at } } a wavelight of 0.037, the resolving power is about 0.17nm. } } } } However, let's decrease the kV to 50kV or place a thicker sample into } } electron beam. The consequence is an increase in scattered electrons, } } which equals less signal above background. Signal is good, background } } noise is bad. If we decrease our kV we get more contrast but less } } resolution, i.e., more amplitude through reduced resolution. } } } } Spatial and temporal resolution are clearly more difficult to discuss in } } this space or my time. Again, sample thickness, accelerating voltage, and } } condenser/objective apertures (and even degree of vacuum) will play } } differing roles in the outcome of resolution. Then the question arises as } } to how you are going to document this resolution. It is visual, } } photographic, or digital/video? } } } } Based on this information, do you have questions? Perhaps this } } information and that provided by others will help with the anwsers. } } However, who gets the grade for effort. Tell us (the members of the } } listserver) what you believe to be the correct answer(s). It is far } } easier to help through thoughtful questions as opposed to spewing out } } everything one knows on a given subject. } } } } Ken } } ______________ } } Ken Tiekotter, Adjunct Professor } } The University of Portland } } Department of Biology } } 5000 N. Willamette Blvd. } } Portland, OR 97203 }
I think I have it this time.. Could someone give me information on the fixative Zamboni's solution? I need to know how it's made when and why it is used over another fixative.
Thanks to all who responded with advice about TEM of B-galactosidase. I was suprised and glad to find out that one can see B-gal reaction product by TEM. Thanks for the references and helpful hints. I'll give it a try.
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
Director: Research Projects (Director, Electron Microscope Facility) Vacancy Number: 12595 Work Unit: Vice President for Research, Life Sciences Consortium Location: University Park Campus Grade: 24 Exempt
Responsible for enabling faculty to design and implement protocols using transmission and scanning electron microscopes, as well as image and analyze biological and nonbiological materials. Responsible for the maintenance and operation of light microscopes, digital camera interfaces for light and EM microscopy, computers and sample preparation equipment. Train EM personnel and researchers in sample preparation techniques and the efficient operation of instrumentation; teach laboratory courses; develop fee structure; and participate in the development of instrumentation grant proposals. Requires a Master's degree (Ph.D. preferred) or equivalent in Biology, Biochemistry, Molecular Biology or related field, plus two years of previous experience in electron microscopy and two years of a research program. Computer and interpersonal skills required.
Pennsylvania State University Penn State is committed to affirmative action, equal opportunity and the diversity of the workforce. Application deadline is March 11, 2002.
Please email or FAX resume and cover letter to:
Judith Burns, Mgr. STF SVCS, jeb2-at-psu.edu The Life Sciences Consortium 519 Wartik Lab University Park, Pa 16802 FAX: (814) 863-1357 -- Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
Zamboni fixative was initially described in an abstract (Zamboni L, DeMartino, 1967, Buffered picric acid-formaldehyde: a new, rapid fixative for electron microscopy, J Cell Biol 35:148A). It was considered particularly useful for fixing sperm. However, the abstract gave no details about how to make it. A more complete description came out in Stefanini M, De Martino C, Zamboni l, 1967, Fixation of ejaculated spermatozoa for electron microscopy, Nature 216:173-174, which you can check for the details.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
--On Wednesday, February 20, 2002 8:45 AM -0500 kathy lowe {kjl226-at-vt.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 think I have it this time.. } Could someone give me information on the fixative Zamboni's solution? I } need to know how it's made when and why it is used over another fixative. } } Thank you. } Kathy
I have a need to coat 37-mm teflon filters reproducibly, with a uniform, thin (few hundred angstroms) carbon film. The primary objective is not SEM analysis, but to achieve the most uniform coating over the entire filter area. I am hoping that someone more knowledgeable and more experienced can suggest the best type of coater for this purpose. Would one expect a difference in uniformity or reproducibility using carbon yarn vs. carbon rod vs. carbon sputter electrode?
Thank you for all suggestions.
****************************************************** Robert Willis, Ph.D. Principal Scientist ManTech Environmental Technology, Inc. P.O. Box 12313 Research Triangle Park, NC 27709-2313 Tel: 919-541-2809 Fax: 919-541-3566 willis.robert-at-epa.gov ******************************************************
Please could you bring the following job advert to the attention of potentially interested people,
Thanks in advance,
Stu
------------------------------------------------------------------------------------ Electron Microbeam Laboratories - Support Assistant
The Department of Earth Sciences, Bristol UK, is seeking a support assistant to work in its electron microbeam laboratories. The post is a fixed term 12-month appointment. The department is re-equipping this analytical area as part of a recent award. The appointee will assist in the demanding role of maintaining a fully operational range of analytical services whilst assisting with the procurement, installation and commissioning of new equipment in new purpose built laboratories.
The primary role of the successful applicant will be to maintain equipment, and to train and supervise researchers and students in all aspects of electron microprobe and scanning electron microscope techniques. Additionally the post affords the opportunity to become familiar with modern state-of-the-art equipment during installation.
The electron microbeam laboratories form one of several analytical facilities within the EU Large Scale Geochemical Facility in the department of Earth Sciences. Scientists from European countries visit Bristol for periods of typically one to two weeks to collect data from a variety of instruments on a vast range of research topics. It is expected that the appointee will oversee some of this external work.
The successful applicant will have a background in an Earth Science discipline and experience in wavelength dispersive electron microprobe analysis. She/He will also possess excellent communications skills. Further experience of SEM techniques and sample preparation would be an advantage.
Approximate start date - July 2002. Applications by CV, covering letter and names and addresses of three referees to Dr Stuart Kearns at the address below by 9am, 15th March 2002. For informal enquiries and further particulars, please contact Dr. S.L.Kearns (stuart.kearns-at-bristol.ac.uk) Dept. Earth Sciences, Queens Road, University of Bristol, Bristol, UK, BS8 1RJ ------------------------------------------------------------------- Stuart Kearns Electron Microbeam Laboratories Department of Earth Sciences University of Bristol Queens Road Bristol BS8 1RJ UK tel: (0)117 954 5435 fax: (0)117 925 3385 e-mail: Stuart.Kearns-at-bristol.ac.uk http://www.gly.bris.ac.uk http://eugf.gly.bris.ac.uk --------------------------------------------
You don't know how much I truly appreciate your time and answer(also to Gary). Since I am not a OLM person, much of the knowledge relates to that is coming from textbook and misconception, especially for Q1.
Ken Tiekotter wrote:
} *You must specify TEM or SEM or STEM or ESEM. You cannot ask these } questions as to how they relate solely to "EM", as the answers relating to } physical design and the physics associated with their operation are unique } to each instrument. } } You state "better depth of field/focus". Yes, in the SEM the scanning } probe instruments, depth of field/focus is important. However, depth of } field in a 90nm section for TEM is, as you can imagine, extremely small, } hence the advantage of accelerating voltage.
Sorry, I simplified too much. However, TEM does have large depth of FOCUS.
} As for the disadvantages of EM as being labor intensive, have you any } knowledge of LM preparation or the difference between TEM and SEM } preparation? SEM preparation is a walk in the park compared to the rigors } necessary to preserve, examine, and document ultrastrutural resolution at } 0.1nm. } } ... much deleted... } } As for instrument expense, look into a multi-photon, scanning laser } confocal OLM microscope at $400k to $600K!
I did not know OLM can cost that much and how labor intensive it can be. Please excuse my ignorance. I believe my answer is from some textbook. Shouldn't this be the very reason that the student should ask YOUR opinion rather than just the textbook?
} } If I ask all the light microscopist, how many of your tasks can be } } simply replaced with electron microscopy - presume you have enough } } funding and man power? } } Toolbox...more expensive toys do not necessarily answer the simplest } questions
Excellent analogy. That is why I think Q1 maybe misleading. You can not really compare the advantage and disadvantage directly. Most of the task by OLM can not replaced by EM, or vice versa. Just like you can not ask "what is the advantage to use a power scroll driver other than a hammer?". However, as I suggested earlier, you may list the general advantage/disadvantage of each individual tools in the toolbox.
} Do you not agree there is a limit to the resolution of the OLM regardless } of wavelength or with respect to wavelength? If the OLM resolution were } limitless with respect to wavelength, why bother with the TEM?
Thank you to point that out. I don't know exactly what you meant on the limitation
of OLM, but one thing I can think of is the X-ray microscopy(if you still consider
X-ray is a light). Whatever your reason is, not only we should "bother" with TEM, but also we should "bother" with "OLM". If you are seeking for a simpler answer to Q2, it could be "generally the shorter the wavelength the better the microscope resolution. However it may not be true when the wavelength is too short". Wouldn't what make the answer better?
} } ... much deleted ...
I have learned so much from Q1 and Q2. For Q3, I think I know generally what "temporal resolution". But I am still seeking answers to what is means in EM. Thanks to Gary, I start to understand, but not good enough to teach another person. If no one answers, I can do some more research myself.
Finally, I have some general comment: I believe there are three groups of people when facing a certain question :
First group is those who don't know the answer. Second group is those who are not sure about the answer Third group is those who know the correct answer. Of course there is another group who doesn't care about the answer.
In the first group. there are who don't admit, or unwilling to ask, (e.g. hopefully none) and who are not afraid to admit and willing to ask(e.g. the student and hopefully meself) In the second group, there are who don't wish to discuss and those who wish to discuss and seeking the right answer(e.g. us) In the third group, there are who are not willing to teach and who are willing to teach(e.g. Ken, Gary, and many more out there...)
So thank you so much to those who are interested in these discussions. I hope you learn as much as I do.
Yours sincerely, Xudong
********************************************************************* Confucius Says: If you know, you do. If you don't know, you don't. Don't be ashamed to ask. *********************************************************************
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Listers, In relation to this, resolution claims by original equipment manufacturers for SEMs are usually based on images of a gold on carbon sample. This is an "ideal" sample. However, few of us have ideal samples. Does anyone have information on realistic resolutions to expect from a FEG-SEM as compared with a standard SEM with tungsten filament for the following sample types?
a) dry, coated biological(or low density polymer) sample - low/medium topography b) hydrated sample such as polymer or collagen gels prepared using cryo-SEM c) hydrated nanotubes or vesicles using cryo-SEM d) pure metal samples looking for grain boundaries- uncoated
I understand that FEG should give 3-5 times better resolution at low kV than standard gun but do not know how to relate that to real life samples. Information such as working distance and kV used as well as magnificaitons when determining the resolution would be of interest. Any reasonable guess would be appreciated. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907
On Wednesday, February 20, 2002 3:33 AM, Chris Jeffree {c.jeffree-at-ed.ac.uk} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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At 09:48 AM 2/20/2002, you wrote:
} I have learned so much from Q1 and Q2. For Q3, I think I know generally } what "temporal resolution". But I am still seeking answers to what is means } in EM. Thanks to Gary, I start to understand, but not good enough to } teach another person. If no one answers, I can do some more research myself.
Temporal has to do with time. Thus, temporal resolution is the ability to resolve or capture an image when the specimen is moving. With LM, a video camera can do this as would multiple snaps with a film camera. The reason this works is that the image is immediately captured and thus, specimen motion is less of an issue (all other things being equal of course).
For SEM, temporal resolution is a problem. Imagine a micromachine moving at say 20 Hz. And we want to capture that movement in the SEM at 5KX. How long does it take to slow scan a frame? Shortest on my system is about 30 Seconds. So the object is moving at a 50mS rate but I can only capture one image every 30 Seconds. Bad temporal resolution. In slow scan mode, even the fastest of the slow scan rates, I can't see the machine move. Under LM, I could, if I could get 5KX mag. Perhaps a confocal would do this. But irrespective, this is the basic idea.
A solution? Don't have the machine continuously move. Step it in discrete amounts and capture each position. Then, put them all together into an avi or Quicktime file.
Another option is to video record the RS-170 out of my SEM in partial field. This is a faster scan rate which is buffered and output as a high rez RS-170 field. Other systems can likely do this too.
A graduate student is interested in looking at viability of freshly collected pollens from his research plants. Is there a quick method to do this instead of looking at germination ? Is there any vital stain or other dyes available ? We do have access to epiflourescence microscope.
Thanks in advance.
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
I've seen Gold on Carbon and Tin on Carbon samples. Even these vary from specimen to specimen, how they were stored, how long they were stored, etc.
The main advantage of FESEM is brightness, in my view. Arguably, the Schottky FE is more stable than a cold FE. Be that as it may, brightness is a given I'm sure. Resolution is, I believe, based on three factors: 1) specimen contrast, 2) electron optics performance, and 3) sample volume limitations.
Number 1 is why the makers like Gold on Carbon-- huge contrast. Contrast values from 1 to 1E-3 can have related probe sizes from 25A to 25,000A.
The electron optics are really critical. Just their basic inherent design. For example, I work with two FESEMs, both of which use the same Schottky FE gun. From the bottom of the gun assembly on down the column, all else is quite different. One SEM uses a conical lens design while the other is a flat lens design. For the same magnification and WD, the conical lens SEM will at 2KV produce basically the same quality image as the flat lens SEM does at 10KV. On a really good day, maybe at 5-6KV. The final aperture in the conical system is 70u while a 100u aperture is used in the flat lens system. I doubt that this makes much difference overall.
I don't feel qualified to discuss sample volume limitations. Perhaps others can jump in on this facet.
Based on your specimens, it seems that the first limiting factor is specimen contrast. When one thinks that the machine is faulty, it may actually be the specimen which is limiting the resolution.
BTW, both FESEMs are tested with Gold on Carbon for resolution. Conical is done at 5KV, 70u, 4mm, 200KX. Flat guy is done at 10KV, 100u, 4mm, 200KX. Pretty much the same results between the two. The killer for astigmatism turns out to be the apertures (dirty) while overall poor resolution (for good specimens) is a dirty scan coil liner.
gary
At 09:51 AM 2/20/2002, you wrote: } Listers, } In relation to this, resolution claims by original equipment } manufacturers for SEMs are usually based on images of a gold on carbon } sample. This is an "ideal" sample. However, few of us have ideal } samples. Does anyone have information on realistic resolutions to expect } from a FEG-SEM as compared with a standard SEM with tungsten filament for } the following sample types? } } a) dry, coated biological(or low density polymer) sample - low/medium } topography } b) hydrated sample such as polymer or collagen gels prepared using cryo-SEM } c) hydrated nanotubes or vesicles using cryo-SEM } d) pure metal samples looking for grain boundaries- uncoated } } I understand that FEG should give 3-5 times better resolution at low kV } than standard gun but do not know how to relate that to real life } samples. Information such as working distance and kV used as well as } magnificaitons when determining the resolution would be of interest. Any } reasonable guess would be appreciated. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: } dsherman-at-purdue.edu } S-052 Whistler Building } West Lafayette, IN 47907
} Gary Gaugler wrote: } } } The URLs are not the answer. What is at the URLs is. You have } } to read the material at the URLs to compose an answer for yourself. } } Better still, look up the dictionary definition of "temporal" and } } then read the material at the URLs. If you still don't get it, } } let me know. } } I don't think I have probelm understand the meaning of "temporal } resolution" literally. If I am right, our limited eye "temporal resolution" } allow us to see fast changing stationary pictures(24 frames/sec) as Hollywood } movies. } Sure I have to read the content on URLs. But the content does not provide a } direct answer. Now I can short of figure out what it means with SEM. } Is that means how well you can see a moving sample(MEMS) } vibrating at high frequency in a SEM?
[snip]
I think you have reached the point where one should be able to put the pieces together in a coherent, concise form and answer all of the questions. And in addition, be able to discuss the nuances of LM, SEM and TEM in general and with specifics.
Given the number and types of factors surrounding the questions, and those already disclosed, some on-line research and bookwork should do the job nicely. I'm pretty sure it would. Don't you think so?
Just before you send this, I think I figured out myself too. I am a TEM person, so the concept of observing moving object never come to me. Even so, TEM never fail to capture any move. So I figure out it must be something to do with the scan system. Does the fastest scan rate(TV rate ~ 50Hz) has anything to do with the temporal resolution limit of 50-100Hz in SEM?
I don't think there is any meaning to talk about "temporal resolution" in stationary beam microscopes. Should Q3 be better asked as "How is the temporal resolution produced in a scanning microcope(including scanning light microscope)?" rather than "electron microscope"?
By the way, what is the official unit for "temporal resolution?" Hz? or sec? Well, thank you so much.
Gary Gaugler wrote:
} At 09:48 AM 2/20/2002, you wrote: } } } I have learned so much from Q1 and Q2. For Q3, I think I know generally } } what "temporal resolution". But I am still seeking answers to what is means } } in EM. Thanks to Gary, I start to understand, but not good enough to } } teach another person. If no one answers, I can do some more research myself. } } Temporal has to do with time. Thus, temporal resolution is } the ability to resolve or capture an image when the specimen } is moving. With LM, a video camera can do this as } would multiple snaps with a film camera. The reason } this works is that the image is immediately captured and } thus, specimen motion is less of an issue (all other things } being equal of course). } } For SEM, temporal resolution is a problem. Imagine a } micromachine moving at say 20 Hz. And we want to capture } that movement in the SEM at 5KX. How long does it } take to slow scan a frame? Shortest on my system is } about 30 Seconds. So the object is moving at a 50mS } rate but I can only capture one image every 30 Seconds. } Bad temporal resolution. In slow scan mode, even the } fastest of the slow scan rates, I can't see the machine } move. Under LM, I could, if I could get 5KX mag. Perhaps } a confocal would do this. But irrespective, this is the basic } idea. } } A solution? Don't have the machine continuously move. } Step it in discrete amounts and capture each position. } Then, put them all together into an avi or Quicktime file. } } Another option is to video record the RS-170 out of } my SEM in partial field. This is a faster scan rate } which is buffered and output as a high rez RS-170 field. } Other systems can likely do this too. } } gary g.
} I think you have reached the point where one should } be able to put the pieces together in a coherent, concise } form and answer all of the questions. And in addition, } be able to discuss the nuances of LM, SEM and TEM } in general and with specifics. } } Given the number and types of factors surrounding the } questions, and those already disclosed, some on-line } research and bookwork should do the job nicely. } I'm pretty sure it would. Don't you think so? } } gary g.
Yes, I am glad I did - BEFORE you posted the right answer. I am also VERY glad that you did not tell me the straight answer at the beginning. You are a great teacher. However, if I am going to teach another person about this, I will correct the question first.
I sent this, in a slightly different form, to the microprobe server, but it doesn't seem to have been broadcast, so sorry to those who receive it twice and to those who aren't interested in this sort of thing.
My EDS quantitation software, like most, I think, allows a choice of valency for Fe expressed as oxide, so that it comes out as either FeO or as Fe2O3.
We habitually choose to express it as FeO, and subsequently use one of the several methods available to recalculate some of the Fe as Fe2O3 as necessary, however, it's not the recalculation that I invite discussion on at this stage.
My question is this:
If I had a perfect, stoichiometric, 100% pure magnetite, Fe3O4, (which formula corresponds to 72.36% Fe, 27.64% O), and analysed it, expressing the result as FeO (which formula corresponds to 77.73% Fe, 22.27% O), what should the result be?
It's easy enough to say OK, the magnetite has 72.36% Fe, and if we express 72.36% Fe as FeO, it's simply 72.36 divided by 0.7773, which is 93.09.
So perfect magnetite should come out as 93.09% FeO.
But is this correct, or are there a few unjustifiable assumptions included?
The reason I want to know is that I'd like to start using a magnetite as the calibration standard for Fe for users to analyse titanomagnetites. They are accustomed to the Fe in their titanomagnetites being expressed as FeO.
I have an Astimex standard magnetite which is stated to contain 31.03%FeO, 68.76%Fe2O3, and 0.20%Cr2O3.
So what should I take as its reference value if all the Fe is expressed as FeO?
All opinions welcome.
cheers
rtch
Dr 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
Heslop-Harrison J & Y . 1970. Evaluation of pollen viability by enzymatically induced fluorescence; intracellular hydrolysis of fluorescein diacetate. Stain
Tech 45:115-20
Heslop-Harrison J, Heslop-Harrison Y, Shivanna KR. 1984. The evaluation of pollen quality; and a further appraisal of the fluorochromatic (FCR) test procedure. Theor Appl Genet 67:367-75
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
} ------------------------------------------------------------------------ } 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 everybody, } } A graduate student is interested in looking at viability of freshly } collected pollens from his research plants. Is there a quick method to do } this instead of looking at germination ? Is there any vital stain or other } dyes available ? We do have access to epiflourescence microscope. } } Thanks in advance. } } Soumitra } } ************************************************************* } Soumitra Ghoshroy Ph.D. } Director, Electron Microscopy Lab } Graduate Faculty, Department of Biology } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } Tel: 505-646-3268 (office), 646-3283 (lab) } Fax: 505-646-3282 } e-mail:sghoshro-at-nmsu.edu } URL:http://confocal.nmsu.edu/eml
AMEN! Ken -- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203
On Wed, 20 Feb 2002, Gary Gaugler 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 } -----------------------------------------------------------------------. } } } At 11:32 AM 2/20/2002, you wrote: } } } } Gary Gaugler wrote: } } } } } The URLs are not the answer. What is at the URLs is. You have } } } to read the material at the URLs to compose an answer for yourself. } } } Better still, look up the dictionary definition of "temporal" and } } } then read the material at the URLs. If you still don't get it, } } } let me know. } } } } I don't think I have probelm understand the meaning of "temporal } } resolution" literally. If I am right, our limited eye "temporal resolution" } } allow us to see fast changing stationary pictures(24 frames/sec) as Hollywood } } movies. } } Sure I have to read the content on URLs. But the content does not provide a } } direct answer. Now I can short of figure out what it means with SEM. } } Is that means how well you can see a moving sample(MEMS) } } vibrating at high frequency in a SEM? } } [snip] } } I think you have reached the point where one should } be able to put the pieces together in a coherent, concise } form and answer all of the questions. And in addition, } be able to discuss the nuances of LM, SEM and TEM } in general and with specifics. } } Given the number and types of factors surrounding the } questions, and those already disclosed, some on-line } research and bookwork should do the job nicely. } I'm pretty sure it would. Don't you think so? } } gary g. } }
Dear all, Does anyone know what became of the materials-l email discussion group that was based at Liverpool (UK). I tried logging on yesterday and got an error message saying that the address listproc-at-liv.ac.uk didn't exist.
Thanks
Ian MacLaren MPI für Metallforschung, Heisenbergstr. 3, 70569 Stuttgart, Germany http://www.mpi-stuttgart.mpg.de/
EC TMR-Network NANOCOMP (1 POST- and 1 DOCTORAL Research Position)
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As part of the NANOCOMP programm
1 POST- and 1 DOCTORAL Research Position
are available (start date: immediately) at the Max-Planck-Institut fr Metallforschung in Stuttgart, Germany
Within the network the Stuttgart research team is responsible for the Microstructural Characterization of Carbon Nanotubes and Carbon Nanotube-based Composites using
* High resolution transmission electron microsocopy, * High spatial resolution electron energy-loss spectroscopy and EDX and * other characterization tools (SEM, AFM, STM, XPS, etc.) The department of Prof. M. Rhle at the MPI fr Metallforschung in Stuttgart offers exceptional transmission electron microscopy instrumentation and expertise.
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The aim of the group at the MPI fr Metallforschung is to develop new methods for the controlled sythesis of carbon nanotubes and related structures. These materials will be employed for novel composite materials exhibiting interesting chemical and physical properties. As a member of the NANOCOMP network you will have the exciting chance to perform research in close European collaboration with Postdocs and PhD students of other leading European teams including regular network meetings, research stays at partner groups and the participation at European conferences.
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Contact (Information and Application)
Prof. Dr. Manfred Rhle Phone: +49-711-689-3520 Ruehle-at-mf.mpg.de
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I wish to digitally save and print images from a Philips 515 SEM. Is there some way to get an output from this SEM that can be saved as a Bitmap or TIFF file, or otherwise digitized and printed?
I'm reaching back here, but isn't magnetite a mix of Fe+2 and Fe+3 and you should get both FeO and Fe2O3?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Thursday, February 21, 2002 7:08 AM To: Microscopy-at-sparc5.microscopy.com
Hi
I sent this, in a slightly different form, to the microprobe server, but it doesn't seem to have been broadcast, so sorry to those who receive it twice and to those who aren't interested in this sort of thing.
My EDS quantitation software, like most, I think, allows a choice of valency for Fe expressed as oxide, so that it comes out as either FeO or as Fe2O3.
We habitually choose to express it as FeO, and subsequently use one of the several methods available to recalculate some of the Fe as Fe2O3 as necessary, however, it's not the recalculation that I invite discussion on at this stage.
My question is this:
If I had a perfect, stoichiometric, 100% pure magnetite, Fe3O4, (which formula corresponds to 72.36% Fe, 27.64% O), and analysed it, expressing the result as FeO (which formula corresponds to 77.73% Fe, 22.27% O), what should the result be?
It's easy enough to say OK, the magnetite has 72.36% Fe, and if we express 72.36% Fe as FeO, it's simply 72.36 divided by 0.7773, which is 93.09.
So perfect magnetite should come out as 93.09% FeO.
But is this correct, or are there a few unjustifiable assumptions included?
The reason I want to know is that I'd like to start using a magnetite as the calibration standard for Fe for users to analyse titanomagnetites. They are accustomed to the Fe in their titanomagnetites being expressed as FeO.
I have an Astimex standard magnetite which is stated to contain 31.03%FeO, 68.76%Fe2O3, and 0.20%Cr2O3.
So what should I take as its reference value if all the Fe is expressed as FeO?
All opinions welcome.
cheers
rtch
Dr 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
} } By the way, what is the official unit for "temporal resolution?" Hz? or sec? } Since Hz is frequency, temporal resolution must be sec (or msec)...
Okay, this helps me understand better. This means that the temporal resolution is not just for vibrating object. Resolution of 1sec means you can not resolve vibration higher than 1Hz, No problem. What does this 1sec mean if you wish to observe an object which is moving in one direction? My definition: you can only capture the motion that is taking more than 1sec to across the screen. For example, your scan rate must be longer than 1sec/frame, you only can capture one position of the object, the following frames catches nothing. So the motion is not resolved. However, if I keep this scan rate, and simply reduce the magnification, say by 10X, now I can capture the motion by capture 10 positions of the object. Now the same motion is resolved. Was the temporal resolution improved? I guess not.
Hi Soumitra, Yes, there is a quick and easy way. I don't have my original protocol, but this one will probably do (from Ruzin's Plant Microtechnique and Microscopy). Dissolve 1 mg of fluorescein diacetate (FDA) in 1 mL of acetone for a stock solution that can be stored at -20C. Then add to pollen grains for a grand total concentration of 1 microgram FDA/mL and incubate for 5 minutes. Observe under blue light (488 nm). Living cells will fluoresce yellow/green. My pollen was usually in germination medium at the time so that is into what I added the FDA. I'll cross-check this with my old files and let you know if there are any significant differences. Good luck. 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 www.baumlab.org
Dear Debby, I was just using a new cold-FESEM at another department at our university and comparing the results with the photos from our ten-year-old, W-filament SEM on coated ceramic samples with tiny pores. The results were that we could take photos at 200KX on the FESEM with the same clarity as 20KX on our older SEM. They haven't used the cryo much yet, but the resolution seems to be about ten times. We could easily resolve the thin gold-palladium coating. We tried a little 5 Kv work on the FESEM and we could still get a good picture at 150KX, so I think resolution is at least five to ten times that of a W-filament SEM. At 12:51 PM 2/20/02 -0500, you wrote: } } Listers, } In relation to this, resolution claims by original equipment manufacturers for SEMs are usually based on images of a gold on carbon sample. This is an "ideal" sample. However, few of us have ideal samples. Does anyone have information on realistic resolutions to expect from a FEG-SEM as compared with a standard SEM with tungsten filament for the following sample types? } } a) dry, coated biological(or low density polymer) sample - low/medium topography } b) hydrated sample such as polymer or collagen gels prepared using cryo-SEM } c) hydrated nanotubes or vesicles using cryo-SEM } d) pure metal samples looking for grain boundaries- uncoated } } I understand that FEG should give 3-5 times better resolution at low kV than standard gun but do not know how to relate that to real life samples. Information such as working distance and kV used as well as magnificaitons when determining the resolution would be of interest. Any reasonable guess would be appreciated. } Debby
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-interchange.ubc.ca
} From alan.thew-at-liverpool.ac.uk Thu Feb 21 13:48:29 2002 } Date: Thu, 21 Feb 2002 18:51:45 +0000 (GMT Standard Time) } From: Alan Thew {Alan.Thew-at-liverpool.ac.uk} } Reply-To: U of Liverpool List Admin {list.admin-at-liverpool.ac.uk} } To: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} } cc: U of Liverpool List Admin {list.admin-at-liverpool.ac.uk} } Subject: Re: is materials-l down? } } On Thu, 21 Feb 2002, Jim Quinn wrote: } } } } } Alan - } } } } Is "material-l" down on your listserv? } } The right name is } } materials-at-liverpool.ac.uk } } but } } materials-l-at-liverpool.ac.uk } } still works... my test message just went out... } } } } thanks and regards, } } } } JQuinn } } } Alan Thew }
} From owner-MATERIALS-at-LISTSERV.LIV.AC.UK Thu Feb 21 13:49:23 2002 } Date: Thu, 21 Feb 2002 18:52:31 +0000 } From: "L-Soft list server at U of Liverpool (1.8d)" } {LISTSERV-at-liverpool.ac.uk} } Subject: Information about the MATERIALS list } To: jquinn-at-www.matscieng.sunysb.edu } X-LSV-ListID: MATERIALS } } There is no information file for the MATERIALS list. Here is a copy of } the list "header," which usually contains a short description of the } purpose of the list, although its main purpose is to define various list } configuration options, also called "keywords." If you have any question } about the MATERIALS list, write to the list owners at the generic } address: } } MATERIALS-request-at-LISTSERV.LIV.AC.UK } } * } * Materials Science and Engineering list } * } * Review= owner Subscription= open,confirm } * Send= private } * Notify= Yes Reply-to= List,Respect Files= No } * Validate= Yes } * Mail-Via= Direct } * Default-Options=REPRO } * Auto-Delete=Yes,Semi-Auto,Delay(4),Max(30) } * Sizelim= 1000 } * Attachments = No } * Language = NOHTML } * } * Notebook= Yes,/listserv/notebooks/m/materials,Monthly,public } * } * {3 lines hidden} } * }
Sorry, I realize I should be more specific. We are looking for an independent service provider that can service the actual microscope. We are having issues with our microscopes and wanted to know if anyone used an independent service provider for this kind of work, or if anyone knows of someone that fix microscopes.
Any info would be greatly appreciated.
Thanks, Kathy
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} I understand that FEG should give 3-5 times better } resolution at low kV } than standard gun but do not know how to relate that to real } life samples. } Information such as working distance and kV used as well as } magnificaitons } when determining the resolution would be of interest. Any } reasonable guess } would be appreciated. } } Debby
I'll talk about dentin, which when dehydrated is mostly a porous mineral, hydroxyapatite.
For not coated dehydrated dentin in low voltage or low vacuum modes I can use magnifications up to 10k. For not coated hydrated dentin in wet mode - up to 20-40k. For coated with gold-palladium - up to 100-150k. Image of gold on carbon standard is almost as good in wet mode as in conventional mode at 100k. Ceramics I have observed mostly behave similar.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
We have a few spare parts for an ESCA Quantum 2000. Stages, gaskets, etc. } Please e-mail at briemeng-at-hotmail.com
_________________________________________________________________ MSN Photos is the easiest way to share and print your photos: http://photos.msn.com/support/worldwide.aspx
Sorry for those repetitions folks, but I think that this reprint is an opportunity for any who are new to older, histotechnique compendia. I don't know an Krieger, but I have purchased the book, and I use it often.
Highly recommended for personal or library.
Regards,
Fred Monson
} ---------- } From: Krieger } Sent: Thursday, February 21, 2002 5:18 PM } To: Monson, Frederick C. } Subject: Re: Gray Available? } } BOOK #: 202473 ISBN #: 0-88275-247-2 } } AUTHOR: GRAY } } TITLE: MICROTOMIST'S FORMULARY AND GUIDE } } PRICE: 84.50 REFRL/ARRGM TYPE: CLOTH } } Shipping $5.00 UPS } } AT PRSENT WE ARE UPDATING OUR LISTS AND PORTIONS OF LOOKUP ARE OFFLINE. } } THE MICROTOMIST'S FORMULARY AND GUIDE } Peter Gray, } 0-88275-247-2 } } Pages: 808, Binding: Cloth, } } Description: } This is a known and recognized source reference work. The book includes a } treatise on the art of making microscopic slides from biological } specimens, } as well as a classified list of the formulas and techniques used in this } art. } }
I've been hearing that the Coolscan 8000 was pulled from distribution. Mostly (??) due to bad LED and CCD mating.
From recent research, if you want to scan from 35mm on up to TEM negs, look at the Minolta Dimage Scan Multi Pro. it is a 4.8D, 4800dpi 35mm, 3200dpi MF scanner with fluorescent lamp light source (very good). It interfaces via Ultra SCSI or Firewire. Supposedly, only Win2K and WinME work with it on the PC.
TriState Camera (NYC) lists this unit at $2749 vs. the MSRP of $2999. I have not tried it but am getting itchy.
gary g.
At 11:49 AM 10/25/2001, you wrote:
} Rick, } Many of our customers are very happy with the 8000ED. } I have looked into putting TEM negs into it and it could be } done by modifying one of the 120/220 film holders. } The holders have a raised lip to keep the film in the } channel. I believe these could be removed and the film could then } just extend out past the scan opening. I have not been able to try } this as demand for the scanner has been very high. } Another excellent scanner for TEM negs is the Agfa } T2500 Duoscan. While lower in resolution(2500dpi optical) it has } a glassless carrier design that will enable scanning of an entire } TEM negative. It also will scan reflective originals. } } George
Sorry, in my previous posting I confused things a bit with talk of standards, please let me attempt to rephrase the question:
Let's say you're doing mineral analyses, everything nicely standardised, Fe being expressed all as FeO, with the oxygen not analysed but calculated from the Fe analysis with an assumed valency for Fe of 2.
Then you hit a pure, stoichiometric magnetite, formula Fe3O4 (72.36% Fe, 27.64% O).
Because you're expressing Fe as FeO, the result will be too low, and will be something like:
FeO 93%
This is the 72.36 multiplied by the formula weight ratio of FeO/Fe (71.85/55.85, ie 1.2865), and comes actually to 93.09%.
Is that all there is to it?
Is that the result that you would expect/accept?
Or does the underestimation of oxygen content which results from the assumption of Fe valency of 2 have a significant effect on the ZAF calculations or on some other part of the calculations involved in the generation of the result?
I'm grateful for the replies received so far, but would you mind posting them on the list as well as to me personally?
cheers
rtch
Dr 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
SEGREGATION´02 1st International Conference Košice, 6th – 7th June 2002
An International Conference marking the 50th anniversary of the establishment of the Metallurgical Faculty at the Technical University in Košice. This conference continues a series of occasional scientific meetings on segregation of impurities and their effect on the applied properties of metal materials which have previously been held at the Department of Materials Science, Faculty of Metallurgy of the Technical University in Košice. Segregation processes occur in the majority of steels from crystallization through to the final technical processing into finished products. This means that they influence material quality and applied properties over the whole range of production technology, including casting, welding, shaping and heat- or chemical-heat treatment. Segregation processes also continue to affect finished products in connection with their external working conditions. The 1st International Conference on Segregation will focus primarily on theoretical questions of phase interfaces and segregation processes as well as their degradation features, in the following areas:  identification methods of segregation processes  structure, models and properties of phase interfaces  dendritic segregation  macrosegregation at solidification  grain boundary segregation, or segregation to other phase interfaces  interaction segregation and precipitation  effect of segregation and precipitation on embrittlement and properties of materials at - casting - welding - forming - heat- or chemical–heat treatment - during exploitation The principal aim of conference is to facilitate exchange of the latest insights and the setting-up of personal and professional contacts. The conference languages Slovak, Czech and English
Conference venue Campus of the Technical University in Košice
Call for papers Both oral presentation and posters are welcome, proposals for which should be submitted with a short abstract ( not exceeding 200 words).
Deadlines:
March 4, 2002 : Abstract deadline March 15, 2002: Acceptance notification date April 15, 2002 : Manuscript due
} Let's say you're doing mineral analyses, everything nicely } standardised, Fe being expressed all as FeO, with the oxygen not } analysed but calculated from the Fe analysis with an assumed valency } for Fe of 2. } } Then you hit a pure, stoichiometric magnetite, formula Fe3O4 } (72.36% Fe, 27.64% O). } } Because you're expressing Fe as FeO, the result will be too } low, and will be something like: } } FeO 93% } } This is the 72.36 multiplied by the formula weight ratio of FeO/Fe } (71.85/55.85, ie 1.2865), and comes actually to 93.09%. } } Is that all there is to it?
As far as I am aware, yes. In the same way, if you measure calcite (CaCO3), the total for oxides (CaO) will be only about 55% (ignoring other problems!). And if you measure pure Fe2O3, then the result expressed as FeO will be only about 90%, so that multiplying by the FeO -} Fe2O3 conversion factor (1.1113) gives 100%, expressed as Fe2O3.
} Is that the result that you would expect/accept?
Yes, given that you are then going to try and calculate what the true oxidation state is from charge balance. When taken back through the formula calculation that should improve your oxide total, as part of the FeO is then expressed as Fe2O3.
} Or does the underestimation of oxygen content which results from the } assumption of Fe valency of 2 have a significant effect on the ZAF } calculations or on some other part of the calculations } involved in the generation of the result?
This is a possibility, since the oxygen will have some effect upon the Fe correction (but not as much as the other way round, if you try and measure oxygen directly). The effect is probably not very significant - try playing with ZAF corrections off-line to gauge how much.
This is a case where measuring oxygen by difference would be more realistic and helpful than measuring oxygen by stoichiometry.
Cheers,
Norman
================================================= Dr. Norman Charnley Department of Earth Sciences University of Oxford Oxford OX1 3PR, UK.
Why do we all test a new SEM befor buying with our own samples ? It's exactly why the gold of carbon sample is so far from our real needs, as Bebby said. It's good at high voltage, but its only real advantage is that all manufacturers use the same test, however the results may vary with the age of the sample etc...
A nice test I do often with students, is to look at an egg shell : outher wall, inner wall, inner membrane, etc, coated or uncoated, at different energies, on a W-SEM and on a FE-SEM etc. It shows that in fact this "resolution" question in the SEM is a false problem. Is the word resolution attapted to what we speek from ? The real question is :
"What do I see, and what am I missing ? "
When peolpe speak from High Resolution SEM, they think always of FE-SEM. With a FE source, all is not done. To perform HR, I must have of course a FE gun, but also a design of the optic optimised for primary energy for a few hundred eV to a few keV (high voltage is always good), and a choice of detectors which allows to separate or mix SE and BSE.
So, the set of parameters I will choose depends of the sample and of which probleme I have to solve. The type of source is one element among a lot : FE or W fil i.e. more or less brightnes, current, chromatic aberration, spot size WD, i.e. more or less depth of field. And what does depth of field mean, whith a 100k X magnification ? WD gives also more or less SE/BSE in the classical ET detector. Primary energy, i.e. more or less interaction volume, (or other said variation in the ratio between surface and volume informations), more less SE/BSE yield. Detector, i.e. the choice between BSE, SE, or a mixing of the two signals, in conjonction with the primary energy.
It's a new thing for a lot of SEM user, to have to precise which detector has been used to perform the image they are showing. Most people are (more or less...) familiar with the WD and kV indication, but realy less with precision about "in lens-SE ", classical (Everhart-Thornley) or BSE detectors. I have made a test with a interresting sample (carbon nanotubes grown on a Co film on a Si (100) single crystal. The same location, at the same magnification at two energies, with the different detectors avaible, and I obtained 8 differents pictures. A closer look leaves 6 really different pictures, with different resolutions, but with different informations too. By the way, it would be nice if the manufacturer could "standardise" the name of their "In Lens", "Throught the lens", "Upper", "SE" detetcors !
And other aspect must be mentionned, which is the ability to perform "good" resolution images of insulators, without coating. The performences at low energy of the HR-SEM is in that case the key element, and the resolution is not the main factor. We have performed images of organic compounds deposed on an teflon (!) film on glass (!). In a classical SEM, or older FE-SEM, we could not observe anything, because charging. With gold coating, at high energy the surface information is lost, and at low energy, the resolution is too poor. In recent HR-SEM from all brand, we could do it, with correct resolution (crystals from 100 to 300 nm at x 100k ), but at 0.5 to 1.5 keV, SE detector, and less enought charging to perform an image (of course without any coating).
It would be interresting to propose a set of "testing samples" of different atomic number combinaisons, light and light, heavy and heavy, light on heavy and vice versa, dry or hydrated, inorganic or organic, which could everyone may himself or obtain easy, to perform resolution/information tests at different energy, WD, detector, etc. Perheps such a inventory soon exists ?
One could take, for exemple : gold on carbon (to continue the tradition) carbone nanotubes dispersed on a heavy metal (gold is expensive, but why not a W foil) CVD silcon can grow with nice crystallite, wiskers, in particular when there are impurities insulators such as Al2O3, glasses, ceramics (Chris Jeffree told me from the inner surface from a broken halogen lamp, i.e. W crystallites "moving" on the glass. Is it right?) catalysers (heavy metals in ceramic powder, arc owen C nanotubes and its catalysr) to test spatial resolution of the BSE detectors in compo mode. etc...
I am not biologist, so I have no idee what could be a good biological test specimen, in dry or cryo mode.
Other idees ? Yes, certainly !
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
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Listers, } In relation to this, resolution claims by original equipment manufacturers for SEMs are usually based on images of a gold on carbon sample. This is an "ideal" sample. However, few of us have ideal samples. Does anyone have information on realistic resolutions to expect from a FEG-SEM as compared with a standard SEM with tungsten filament for the following sample types? } } a) dry, coated biological(or low density polymer) sample - low/medium topography } b) hydrated sample such as polymer or collagen gels prepared using cryo-SEM } c) hydrated nanotubes or vesicles using cryo-SEM } d) pure metal samples looking for grain boundaries- uncoated } } I understand that FEG should give 3-5 times better resolution at low kV than standard gun but do not know how to relate that to real life samples. Information such as working distance and kV used as well as magnificaitons when determining the resolution would be of interest. Any reasonable guess would be appreciated. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } West Lafayette, IN 47907 } } } } On Wednesday, February 20, 2002 3:33 AM, Chris Jeffree {c.jeffree-at-ed.ac.uk} 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 } } -----------------------------------------------------------------------. } } } } } } Shorter wavelength=greater resolution may be the general rule for all } } imaging systems using waves, but } } in SEMs that is far from the whole story, and no commercial TEM comes } } anywhere close to the theoretical resolution limit. TEM resolving } } power is spoiled by lens aberrations. I remember a recent paper which } } described a way of neutralising spherical aberrations in a TEM using } } an electron mirror. } } } } Early SEM guns and optics were designed for 30 kV, and were virtually } } unusable at 1kV. SEM development since the 60s has been about } } emitters, guns and optics. Now, with FEGSEMs we can see that although } } the fundamental rules still apply, major improvements in overall } } performance at low kV have been possible by using near-monochromatic } } emitters. Resolution in uncoated light-element specimens is now seen } } to be a trade-off between kV/wavelength and beam interaction volume. } } In the surface of a potato starch grain, starch crystal edges can be } } seen at 1 or 2kV, but these are obliterated at 5 or 10 kV as the beam } } interaction volume grows. Resolution in TEM is also a function of } } contrast, which is poorer at high kV, which is why some biological } } TEMs are designed with long working distance optics, trading some } } resolving power to buy higher contrast. EM instruments have greater } } theoretical RESOLVING POWER at higher kV, but in practice the } } RESOLUTION can be poorer. Resolving power is an instrument property. } } Resolution can be a specimen or image property. The two do not always } } coincide. } } } } Chris } } } } } Let's see. In SEM, resolution is inversely proportional to } } wavelength } } } of the electrons. Shorter wavelength, greater resolution, in } } general. } } } Wavelength of electrons is inversely proportional to energy. Higher } } } energy, shorter wavelength. So, higher voltage/energy, higher } } } resolution (more resolving power). My old SX-40 brochure says } } } that it had a resolution of 60A. Newer SEMs are better than that. } } } My SEM is newer than the SX-40 and is supposed to have a } } } resolution of about 40A. I guess so, but I can't measure it. } } } I tried but failed. I get about 120-180A at 15KV. And that's } } } an eyeball guess. I cannot find a perfectly sharp edge! } } } } } } } }
I am looking for SEM services in the Western USA that have high resolution (FE) SEM capabilities (and experience) of integrated circuit samples. Planar, oblique, and cross sections of IC components would be necessary.
We generally have a relatively low volume of samples. Does anyone know of a resource directory of services such as this? Any suggestion would be appreciated.
I agree. So even I can take 10 positions of the object to capture the move, I still have no idea how it moves BETWEEN each of these 10 step. Therefore the temporal resolution does not change. The motion is captured at a LOWER resolution. I may be able to intropolate the moving by fitting a curve of these 10 positions. But that's not what machine tells us. In another word, it's our own judgement beyoung machine resolution.
Now here is another question. For a vibrating object, reduce magnification obviously won't catch the motion. But by blanking the beam periodically(as suggested by Gary and NASA JPL lab), a crips image of the vibrating object can be resolved. Or by adjusting the blanking frequency, it can be seen in a slow motion. Does that mean the instrument's temporal resolution is improved?
Is taking a crisp still picture of a fast moving object the same thing as catching the actual movement of that object?
Michael O'Keefe wrote:
} I guess not, too. } At both mags you can tell that the object moved and by how much (if you had a larger } field of view for the first case, it would tell you how much). } My guess would be that a movement would be "temporally resolved" if you could capture } two sucessive frames showing the minimum possible change. For movement, I guess that } that would be a translation of one atomic distance or bond-length. } } Xudong Fan 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 } } -----------------------------------------------------------------------. } } } } Michael O'Keefe wrote: } } } } } } By the way, what is the official unit for "temporal resolution?" Hz? or sec? } } } Since Hz is frequency, temporal resolution must be sec (or msec)... } } } } Okay, this helps me understand better. This means that the temporal resolution is } } not just } } for vibrating object. Resolution of 1sec means you can not resolve vibration higher } } than 1Hz, No problem. } } What does this 1sec mean if you wish to observe an object which is moving in one } } direction? } } My definition: you can only capture the motion that is taking more than 1sec to } } across the } } screen. For example, your scan rate must be longer than 1sec/frame, you only can } } capture one } } position of the object, the following frames catches nothing. So the motion is not } } resolved. } } However, if I keep this scan rate, and simply reduce the magnification, say by 10X, } } now } } I can capture the motion by capture 10 positions of the object. Now the same motion } } is resolved. } } Was the temporal resolution improved? I guess not.
Try this URL for a non-compreshensive list of commerical sites.
http://www.amc.anl.gov/Docs/NonANL/ComSites.html
Nestor
Your Friendly Neighborhood SysOp
===========================================
} } } I am looking for SEM services in the Western USA that have high } resolution (FE) SEM capabilities (and experience) of integrated } circuit samples. Planar, oblique, and cross sections of IC } components would be necessary. } } We generally have a relatively low volume of samples. Does anyone } know of a resource directory of services such as this? Any } suggestion would be appreciated. } } Thank you } } Curtis Olson }
Chipping in my 2 cents worth, "The Microtomist's Formulary and Guide" is the proverbial gold mine of information.
"Monson, Frederick C." wrote:
} Sorry for those repetitions folks, but I think that this reprint is an } opportunity for any who are new to older, histotechnique compendia. I don't } know an Krieger, but I have purchased the book, and I use it often. } } Highly recommended for personal or library. } } Regards, } } Fred Monson } } } ---------- } } From: Krieger } } Sent: Thursday, February 21, 2002 5:18 PM } } To: Monson, Frederick C. } } Subject: Re: Gray Available? } } } } BOOK #: 202473 ISBN #: 0-88275-247-2 } } } } AUTHOR: GRAY } } } } TITLE: MICROTOMIST'S FORMULARY AND GUIDE } } } } PRICE: 84.50 REFRL/ARRGM TYPE: CLOTH } } } } Shipping $5.00 UPS } } } } AT PRSENT WE ARE UPDATING OUR LISTS AND PORTIONS OF LOOKUP ARE OFFLINE. } } } } THE MICROTOMIST'S FORMULARY AND GUIDE } } Peter Gray, } } 0-88275-247-2 } } } } Pages: 808, Binding: Cloth, } } } } Description: } } This is a known and recognized source reference work. The book includes a } } treatise on the art of making microscopic slides from biological } } specimens, as well as a classified list of the formulas and techniques used } in this } } art.
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 **********************************************
This has just appeared on the American Society for Microbiology's listserver; I'm forwarding it because it may interest many on this list also.
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Susan Musante Manager, Education Programs Education Department American Society for Microbiology 1752 N Street, N.W. Washington, D.C. 20036-2904 phone: 202-942-9282 fax: 202-942-9329 smusante-at-asmusa.org http://www.asmusa.org http://www.microbelibrary.org
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Please do see what you can find out. Check out the Minolta too. It is advertised as being able to directly handle TEM negs. It has some sort of adjustable mask affair.
The unit has good specs and good reviews. Price is not bad. But does it work?
gary
At 05:53 AM 2/22/2002, you wrote: } Gary, } I had not heard that about the 8000. They have been shipping } slowly but } steadily. We receive one every 3-5 weeks. } I am attending the PMA show starting this Sunday and I am meeting } with } Nikon the same day. I'll see what I can find out. } } You are probably aware but keep in mind that the 8000 will not } scan an } entire 3.25 x 4 TEM neg at one time. Max scan width is only 56.9mm to } 63.5mm depending on the film holder used. } } Thanks!! } } George } George Laing } National Graphic Supply } v:(800) 223-7130 x3109 } f:(800) 832-2205 } email: scisales-at-ngscorp.com } -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Thursday, February 21, 2002 7:00 PM } To: scisales-at-ngscorp.com } Cc: MSA listserver } Subject: Nikon Coolscan 8000ED - option } } } I've been hearing that the Coolscan 8000 was } pulled from distribution. Mostly (??) due to } bad LED and CCD mating. } } From recent research, if you want to scan from 35mm } on up to TEM negs, look at the Minolta Dimage Scan Multi Pro. } it is a 4.8D, 4800dpi 35mm, 3200dpi MF scanner with } fluorescent lamp light source (very good). It interfaces } via Ultra SCSI or Firewire. Supposedly, only Win2K and } WinME work with it on the PC. } } TriState Camera (NYC) lists this unit at $2749 vs. the } MSRP of $2999. I have not tried it but am getting itchy. } } gary g. } } } At 11:49 AM 10/25/2001, you wrote: } } } Rick, } } Many of our customers are very happy with the 8000ED. } } I have looked into putting TEM negs into it and it could be } } done by modifying one of the 120/220 film holders. } } The holders have a raised lip to keep the film in the } } channel. I believe these could be removed and the film could then } } just extend out past the scan opening. I have not been able to try } } this as demand for the scanner has been very high. } } Another excellent scanner for TEM negs is the Agfa } } T2500 Duoscan. While lower in resolution(2500dpi optical) it has } } a glassless carrier design that will enable scanning of an entire } } TEM negative. It also will scan reflective originals. } } } } George
We have a Balzer's 301 Freeze Fracture system available which was upgraded with cryopump and full electronic/digital controls approx 10 years ago. While currently not running (the cryopump needs a rebuild) the system is in otherwise reasonable shape. We wish to dispose of it due to retirement of the major system user and a shift in research direction within the department. Interested parties are asked to contact the sender below for further information.
Richard Harris
Laboratory Supervisor Research Imaging Resources Department of Zoology University of Western Ontario London ON CANADA N6A 5B7 rjharris-at-uwo.ca (519) 661-2111 ext 86780 (519) 661-2014 Fax
Dear Mr. Faerber, I was recently evaluating VP-SEMs and one of the samples I used, which proved to be quite difficult to image well, was dried but uncoated flower material from the sensitive plant. This is a fine pollen, about five microns in diameter, with tiny features on it. One SEM manufacturer never did image the fine features. It was imaged either by low voltage or variable pressure and made a good test. The low atomic number meant poor secondary and backscattered electron yield. The main reason gold on carbon is used is that more SE-1s are imaged, which are quite indicative of the beam diameter, and there is less low-resolution electrons from the beam interaction volume. At 01:47 PM 2/22/02 +0100, you wrote: } } Why do we all test a new SEM befor buying with our own samples ? It's } exactly why the gold of carbon sample is so far from our real needs, as } Bebby said. It's good at high voltage, but its only real advantage is that } all manufacturers use the same test, however the results may vary with the } age of the sample etc... } } A nice test I do often with students, is to look at an egg shell : outher } wall, inner wall, inner membrane, etc, coated or uncoated, at different } energies, on a W-SEM and on a FE-SEM etc. It shows that in fact this } "resolution" question in the SEM is a false problem. Is the word } resolution attapted to what we speek from ? The real question is : } } "What do I see, and what am I missing ? " } } When peolpe speak from High Resolution SEM, they think always of FE-SEM. } With a FE source, all is not done. To perform HR, I must have of course a } FE gun, but also a design of the optic optimised for primary energy for a } few hundred eV to a few keV (high voltage is always good), and a choice of } detectors which allows to separate or mix SE and BSE. } } So, the set of parameters I will choose depends of the sample and of which } probleme I have to solve. The type of source is one element among a lot : } FE or W fil i.e. more or less brightnes, current, chromatic } aberration, spot size } WD, i.e. more or less depth of field. And what does depth of field } mean, whith a 100k X magnification ? WD gives also more or less SE/BSE in } the classical ET detector. } Primary energy, i.e. more or less interaction volume, (or other } said variation in the ratio between surface and volume informations), more } less SE/BSE yield. } Detector, i.e. the choice between BSE, SE, or a mixing of the two } signals, in conjonction with the primary energy. } } It's a new thing for a lot of SEM user, to have to precise which detector } has been used to perform the image they are showing. Most people are (more } or less...) familiar with the WD and kV indication, but realy less with } precision about "in lens-SE ", classical (Everhart-Thornley) or BSE } detectors. I have made a test with a interresting sample (carbon nanotubes } grown on a Co film on a Si (100) single crystal. The same location, at the } same magnification at two energies, with the different detectors avaible, } and I obtained 8 differents pictures. A closer look leaves 6 really } different pictures, with different resolutions, but with different } informations too. By the way, it would be nice if the manufacturer could } "standardise" the name of their "In Lens", "Throught the lens", "Upper", } "SE" detetcors ! } } And other aspect must be mentionned, which is the ability to perform } "good" resolution images of insulators, without coating. The performences } at low energy of the HR-SEM is in that case the key element, and the } resolution is not the main factor. We have performed images of organic } compounds deposed on an teflon (!) film on glass (!). In a classical SEM, } or older FE-SEM, we could not observe anything, because charging. With } gold coating, at high energy the surface information is lost, and at low } energy, the resolution is too poor. In recent HR-SEM from all brand, we } could do it, with correct resolution (crystals from 100 to 300 nm at x } 100k ), but at 0.5 to 1.5 keV, SE detector, and less enought charging to } perform an image (of course without any coating). } } It would be interresting to propose a set of "testing samples" of } different atomic number combinaisons, light and light, heavy and heavy, } light on heavy and vice versa, dry or hydrated, inorganic or organic, } which could everyone may himself or obtain easy, to perform } resolution/information tests at different energy, WD, detector, etc. } Perheps such a inventory soon exists ? } } One could take, for exemple : } gold on carbon (to continue the tradition) } carbone nanotubes dispersed on a heavy metal (gold is expensive, } but why not a W foil) } CVD silcon can grow with nice crystallite, wiskers, in particular } when there are impurities } insulators such as Al2O3, glasses, ceramics (Chris Jeffree told me } from the inner surface from a broken halogen lamp, i.e. W crystallites } "moving" on the glass. Is it right?) } catalysers (heavy metals in ceramic powder, arc owen C nanotubes } and its catalysr) to test spatial resolution of the BSE detectors in compo } mode. } etc... } } I am not biologist, so I have no idee what could be a good biological test } specimen, in dry or cryo mode. } } Other idees ? Yes, certainly ! } } } J. Faerber 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-interchange.ubc.ca
Has anyone had experiences with diamond knifes that appear to have a poor quality edge that degrades (breaks) with common safe use?
I have a particular knife (3mm ) that has "dinks" in the edge creating knife marks. I won't comment on the brand unless contacted personally.
I am using the knife on skin and feather barbs, but I've used an older knife as a comparison and it doesn't have the same problem as the newer knife.
Help!!!!!! Tim Quinn University of Kansas Research Assistant Natural History Museum and Biodiversity Research Center Dyche Hall Room 414 Lawrence, KS 6604-2454 785-864-4556 tquinn-at-ku.edu
I have a Philips 515 SEM (with analogue image formation) and I wish to print digital images from it. What are the advantages/disadvantages of using an active vs. a passive slow scan image grabber to acquire images?
Suffering from old-age and hockey fever at the same time is HELL!!! Go North America!!!!
Fred Monson
} ---------- } From: Krieger } Sent: Thursday, February 21, 2002 5:18 PM } To: Monson, Frederick C. } Subject: Re: Gray Available? } } BOOK #: 202473 ISBN #: 0-88275-247-2 } } AUTHOR: GRAY } } TITLE: MICROTOMIST'S FORMULARY AND GUIDE } } PRICE: 84.50 REFRL/ARRGM TYPE: CLOTH } } Shipping $5.00 UPS } } AT PRSENT WE ARE UPDATING OUR LISTS AND PORTIONS OF LOOKUP ARE OFFLINE. } } THE MICROTOMIST'S FORMULARY AND GUIDE } Peter Gray, } 0-88275-247-2 } } Pages: 808, Binding: Cloth, } } Description: } This is a known and recognized source reference work. The book includes a } treatise on the art of making microscopic slides from biological } specimens, } as well as a classified list of the formulas and techniques used in this } art. } } -----Original Message----- } } Date: Friday, February 15, 2002 8:05 AM } Subject: Gray Available? } } } } Do you still have Gray, Microtomist's formulary and Guide? And, is it on } a } } web page? Can't recommend it if it isn't there! } } } } Regards, } } } } FCM } } } } Frederick C. Monson, PhD } } }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (nicholas.welham-at-murdoch.edu.au) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, February 23, 2002 at 19:28:55 ---------------------------------------------------------------------------
Email: nicholas.welham-at-murdoch.edu.au Name: Nick Welham
Organization: Murdoch University
Education: Graduate College
Location: Perth, WA, AUSTRALIA
Question: I have an old finite tube Leitz Ortholux I and have been given about half a dozen infinity objectives which I'd like to use. Does anyone know of a company which makes the appropriate adaptor? regards Nick
I am responding to Kathy Smith's question of independant service providers on the East Coast. We are providing TEM service for the last 35 years. We service Philips, Zeiss, Jeol & Hitachi instruments. We are located close to Philadelphia, PA and cover the entire East Coast. Peter A. Stolzenberg, PESTO Inc., 215-699-6160 FAX 215-699-5275
A quick answer is- advantages of an active system can be divided in two categories.
1) Principal. An active system will create new capabilities, which your SEM doesn't have now. For example, modern PC or a Mac. computer control of applications like electron beam lithography and EDX imaging, which depend on synchronization of either beam blanker or an EDX spectrometer with the e-beam position. Another principal advantage, is that a large portion of SEM electronics (most of the scan generation and video signal processing related circuits) will no longer be needed. This will make an SEM more reliable and less expensive to maintain.
2) Non-principal. An active system will allow much slower scan speed than the SEM scan generator, and much longer integration times (pixel dwell times) for video signal acquisition. That will allow you to acquire higher resolution images easier than with the passive system. Also, active system can better compensate for AC fields related interference, as compared with the passive system.
Active system is more expensive than the passive one. A motorcycle is more expensive than the bicycle. I wouldn't call it a disadvantage.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
This message is made of 100% recycled electrons. ----- Original Message ----- } From: Greg Barclay {gbarclay-at-trinidad.net} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, February 22, 2002 12:56 PM
Greg, Having sold both type of systems for many years now, I'll give you my $.02 worth on the merits of a passive system.
1. While many active systems boast resolutions in excess of 4k x 4k, the time to acquire images at these high resolutions make you think twice. Most SEM manufacturers went to great pains to develop highly accurate, very low drift scan generators to drive their photo crt. Why not take advantage of it? A passively acquired 2k x 2k image takes the same amount of time as the SEM photo scan - typically about one minute. And, on older SEM's like yours, you can select different line dwell speeds to increase collection efficiency.
2. What you see is what you get - that includes on screen data, micron line markers, text, etc. Since the data(which is computed by the SEM) is acquired along with the image, there is no question as to its accuracy of calibration. As long as the SEM is properly calibrated, your passive system is also - automatically. With an active system, its scans have to be calibrated to mimic the ones produced by the SEM. If they are not, then your active system will give you inaccurate results.
3. Passive systems will accept EDS dot mapping signals which you can colorize and save.
4. An active system will require that the SEM to be fitted with some type of external scan interface to enable the computer to drive the scan coils. If it does, fine. If not, one will have to be installed before the system can be used. The passive system simply syncs up to the existing SEM scans. Installation is usually very simple and easy.
Active systems are also very good and will give you added capabilities(beam steering) that a passive system won't, especially when integrated with an EDS system. However, don't accept the advice that a passive system is of any lesser quality or usefulness - it's all in what you intend to use it for now, and in the future.
Gary M. Easton, President Scanners Corporation SEM Service/PC Based Imaging & EDS Sales 90 Aileron Court Suite 6 Westminster, MD 21157 410.857.7633 x102(Voice) 410.857.7636(Fax)
} Greg, } } A quick answer is- advantages of an active system can be divided in two } categories. } } 1) Principal. An active system will create new capabilities, which your SEM } doesn't have now. For example, modern PC or a Mac. computer control of } applications like electron beam lithography and EDX imaging, which depend on } synchronization of either beam blanker or an EDX spectrometer with the } e-beam position. Another principal advantage, is that a large portion of SEM } electronics (most of the scan generation and video signal processing related } circuits) will no longer be needed. This will make an SEM more reliable and } less expensive to maintain. } } 2) Non-principal. An active system will allow much slower scan speed than } the SEM scan generator, and much longer integration times (pixel dwell } times) for video signal acquisition. That will allow you to acquire higher } resolution images easier than with the passive system. Also, active system } can better compensate for AC fields related interference, as compared with } the passive system. } } Active system is more expensive than the passive one. A motorcycle is more } expensive than the bicycle. I wouldn't call it a disadvantage. } } Vitaly Feingold } Scientific Instruments and Applications } 2773 Heath Lane, Duluth GA 30096 } (770)232-7785 ph. } (770)232-1791 fax } (678)467-0012 mobile } } This message is made of 100% recycled electrons. } ----- Original Message ----- } } From: Greg Barclay {gbarclay-at-trinidad.net} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Friday, February 22, 2002 12:56 PM } Subject: Active or Passive Frame Grabber for aqnalogue SEM? } } } } ------------------------------------------------------------------------ } } 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 a Philips 515 SEM (with analogue image formation) and I wish to } print digital images from it. } } What are the advantages/disadvantages of using an active vs. a passive } slow scan image grabber to } } acquire images? } } } } } } } } } }
-----Original Message----- } From: Ascend_jcr [mailto:ascend_jcr-at-att.net] Sent: Sunday, February 24, 2002 10:28 AM To: gbarclay-at-trinidad.net; Microscopy-at-sparc5.microscopy.com
Greg,
A quick answer is- advantages of an active system can be divided in two categories.
1) Principal. An active system will create new capabilities, which your SEM doesn't have now. For example, modern PC or a Mac. computer control of applications like electron beam lithography and EDX imaging, which depend on synchronization of either beam blanker or an EDX spectrometer with the e-beam position. Another principal advantage, is that a large portion of SEM electronics (most of the scan generation and video signal processing related circuits) will no longer be needed. This will make an SEM more reliable and less expensive to maintain.
2) Non-principal. An active system will allow much slower scan speed than the SEM scan generator, and much longer integration times (pixel dwell times) for video signal acquisition. That will allow you to acquire higher resolution images easier than with the passive system. Also, active system can better compensate for AC fields related interference, as compared with the passive system.
Active system is more expensive than the passive one. A motorcycle is more expensive than the bicycle. I wouldn't call it a disadvantage.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
This message is made of 100% recycled electrons. ----- Original Message ----- } From: Greg Barclay {gbarclay-at-trinidad.net} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, February 22, 2002 12:56 PM
I think that an active system does create new capabilities. It allows different aspect ratios and also variable pixel dimensions. Additionally, unless the SEM allows for legends to be turned off, only any active system will capture a pure image (no alpha stuff) so that it can be image processed later on. Many times, the alpha creates anomalies during processing. Sometimes it does not. Without alpha, it is not an issue.
I find that the requisite pixel dwell time is inversely proportional to the pixel dimensions. i.e., the more pixels, the shorter the dwell time. With small pixel dimensions, longer pixel dwell time reduces noise which also happens with larger pixel dimensions but at shorter dwell times. Either reduce noise at small pixel dimensions via longer dwell time or increase pixel dimensions and reduce dwell time. For image processing, I like 3088x2060 at 5uS. This replaces the recording CRT's 1.33 aspect ratio to 1.5--which fits nicely on a 35mm slide. Or, change it to 1.33 and output to 6x4.5cm slides. With an active system, there is an option, with passive, not.
The SEM's scan system is of course necessary to do stig, positioning, etc. Whether the EDS system comes with beam control or not (of course it will), unless the SEM is designed to allow external beam control, one needs schematics and some soldering to gain access to the scan driver amplifiers. But he does not mention EDS requirements, just digital capture. So all of the EDS stuff is irrelevant. If the SEM is designed properly, its imaging system will be sync'd to line frequency. Active systems can do this too. Furthermore, some can add micron markers, other data and text. It is optional for the operator to do so or not.
A passive system is certainly going to be cheaper. If that is an issue, go for a passive system. My system is both active and passive. I don't use the passive mode. Fortunately, my SEM provides slow scan info as RS-170. This is easily framegrabbed to accompany a high resolution active scan. Neat.
BTW, the current Rontec EDS system is active scan. Does a nice job.
gary g.
At 10:32 AM 2/24/2002, you wrote:
} -----Original Message----- } } From: Ascend_jcr [mailto:ascend_jcr-at-att.net] } Sent: Sunday, February 24, 2002 10:28 AM } To: gbarclay-at-trinidad.net; Microscopy-at-sparc5.microscopy.com } Subject: RE: Active or Passive Frame Grabber for aqnalogue SEM? } } } Greg, } } I couldn't help comment on this situation as Mr. Feingold obviously has some } hidden agenda or doesn't grasp the concepts fully. First, an active system } will not create new capabilities. It simply duplicates what your SEM does } with the addition of independant digital sampling. Yes, it can drive the } beam, but rarely better than the original SEM hardware which you will still } need, incidentally for correcting stig, alignments, etc. What he says about } EDS is partially true but most EDS systems (including older ones) come with } beam control packages. The advantage of getting the beam control from the } EDS system is that they provide integrated software to do digital mapping, } linescans, in addition to electron imaging. Just purchasing an archiving } system, limits you to electron imaging: you would then have to undertake a } multi-man-year project to integrate with the EDS spectrometer. } None of the second part of Feingold's response is accurate. Slower dwell } times generally do not improve the image for two reasons: First, is that you } tend to build up charging and/or contamination on a stationary spot (are you } familiar with the dark square left behind by the raster?). Secondly, all } SEMs have a stage drift factor from signififcant to severe. Sitting on a } stationary spot will actually result in a record of the stage drift. This is } why the EDS companies provide elaborate drift compensation programs with } their digital beam control packages. } } A further disadvantage of an active system is that it loses all instrument } information unless it maintains a separate communication line with the SEM } for recording kV, mag, etc. } } Passive systems are far superior for pure image collection applicatons such } as digital image archiving. They retain the microscope information printed } on the data bar. They can achieve higher resolution and in some cases } perform frame averaging which is a for superior method of reducing noise } over point averaging since charge doesn't build up. } } I have access to both types systems in addition to a system provided by Edax } and would be happy to provide more detailed information if you want. Is } Feingold trying to sell an active system? I didn't know there were any left } on the market. } } Regards, } Joe Robinson
I think the tone of the comments about Vitaly Feingold's alleged agenda are a bit snide.
At least Vitaly is sufficiently upfront to include his affiliation in his signature, as should everyone who posts to a list.
IMHO
cheers
rtch
} From: "Ascend_jcr" {ascend_jcr-at-att.net} } To: "Net Gbarclay-at-Trinidad." {gbarclay-at-trinidad.net} } Cc: "microscopy. com Microscopy-at-sparc5." {Microscopy-at-sparc5.microscopy.com} } Subject: FW: Active or Passive Frame Grabber for aqnalogue SEM? } Date: Sun, 24 Feb 2002 10:32:33 -0800 } Importance: Normal
} -------------------------------------------------------------------- } ---- 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 } -------------------------------------------------------------------- } ---. } } } } -----Original Message----- } } From: Ascend_jcr [mailto:ascend_jcr-at-att.net] } Sent: Sunday, February 24, 2002 10:28 AM } To: gbarclay-at-trinidad.net; Microscopy-at-sparc5.microscopy.com } Subject: RE: Active or Passive Frame Grabber for aqnalogue SEM? } } } Greg, } } I couldn't help comment on this situation as Mr. Feingold obviously } has some hidden agenda or doesn't grasp the concepts fully. First, } an active system will not create new capabilities. It simply } duplicates what your SEM does with the addition of independant } digital sampling. Yes, it can drive the beam, but rarely better than } the original SEM hardware which you will still need, incidentally } for correcting stig, alignments, etc. What he says about EDS is } partially true but most EDS systems (including older ones) come with } beam control packages. The advantage of getting the beam control } from the EDS system is that they provide integrated software to do } digital mapping, linescans, in addition to electron imaging. Just } purchasing an archiving system, limits you to electron imaging: you } would then have to undertake a multi-man-year project to integrate } with the EDS spectrometer. None of the second part of Feingold's } response is accurate. Slower dwell times generally do not improve } the image for two reasons: First, is that you tend to build up } charging and/or contamination on a stationary spot (are you familiar } with the dark square left behind by the raster?). Secondly, all SEMs } have a stage drift factor from signififcant to severe. Sitting on a } stationary spot will actually result in a record of the stage drift. } This is why the EDS companies provide elaborate drift compensation } programs with their digital beam control packages. } } A further disadvantage of an active system is that it loses all } instrument information unless it maintains a separate communication } line with the SEM for recording kV, mag, etc. } } Passive systems are far superior for pure image collection } applicatons such as digital image archiving. They retain the } microscope information printed on the data bar. They can achieve } higher resolution and in some cases perform frame averaging which is } a for superior method of reducing noise over point averaging since } charge doesn't build up. } } I have access to both types systems in addition to a system provided } by Edax and would be happy to provide more detailed information if } you want. Is Feingold trying to sell an active system? I didn't know } there were any left on the market. } } Regards, } Joe Robinson } } } -----Original Message----- } } From: Vitaly Feingold [mailto:vitalylazar-at-worldnet.att.net] } Sent: Saturday, February 23, 2002 3:59 PM } To: gbarclay-at-trinidad.net; Microscopy-at-sparc5.microscopy.com } Subject: Re: Active or Passive Frame Grabber for aqnalogue SEM? } } } -------------------------------------------------------------------- } ---- 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 } -------------------------------------------------------------------- } ---. } } } Greg, } } A quick answer is- advantages of an active system can be divided in } two categories. } } 1) Principal. An active system will create new capabilities, which } your SEM doesn't have now. For example, modern PC or a Mac. computer } control of applications like electron beam lithography and EDX } imaging, which depend on synchronization of either beam blanker or } an EDX spectrometer with the e-beam position. Another principal } advantage, is that a large portion of SEM electronics (most of the } scan generation and video signal processing related circuits) will } no longer be needed. This will make an SEM more reliable and less } expensive to maintain. } } 2) Non-principal. An active system will allow much slower scan speed } than the SEM scan generator, and much longer integration times } (pixel dwell times) for video signal acquisition. That will allow } you to acquire higher resolution images easier than with the passive } system. Also, active system can better compensate for AC fields } related interference, as compared with the passive system. } } Active system is more expensive than the passive one. A motorcycle } is more expensive than the bicycle. I wouldn't call it a } disadvantage. } } Vitaly Feingold } Scientific Instruments and Applications } 2773 Heath Lane, Duluth GA 30096 } (770)232-7785 ph. } (770)232-1791 fax } (678)467-0012 mobile } } This message is made of 100% recycled electrons. } ----- Original Message ----- } } From: Greg Barclay {gbarclay-at-trinidad.net} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Friday, February 22, 2002 12:56 PM } Subject: Active or Passive Frame Grabber for aqnalogue SEM? } } } } ------------------------------------------------------------------------ } } 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 a Philips 515 SEM (with analogue image formation) and I wish to } print digital images from it. } } What are the advantages/disadvantages of using an active vs. a passive } slow scan image grabber to } } acquire images? } } } } } } } } Dr 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
Let me put in my 2 cents worth. We had discussed our ADDA off-line before, and perhaps that prompted you to ask your question on the list server.
A SEM works by moving an electron beam across the sample and measuring the interaction between the beam and sample at each point. SEM manufacturers have put in a lot of effort to overcome the problems with electromagnetic lenses, focusing and the like. The result are the high resolution instruments of today. However, until a few years ago, there was no way of recording the images other than through film. The medium of choice very quickly became the Polaroid Instant Film, which although still a chemical process, allowed quick and relatively easy dissemination of the images.
Most SEMs have a special photo mode, where the image is scanned with 1000 or 2000 lines and a total duration of a few minutes.
As Joe said, the digital scan interfaces do not try to replace all the electronics of the microscope that the manufacturers put so much effort in. Each SEM on the market, however, uses a similar chain of electronics. First there is a scan generator which produces a precise voltage ramp that in the end moves the beam across the sample. After that there is a scan amplifier, which amplifies the voltage sweep to the actual voltage needed for the sweep. This is determined by the magnification that is required. There is more electronics for other aspects of the microscope such as tilt and astigmatism correction, etc.
With a passive system, you leave the entire chain intact and "only" digitize the signal as it is produced by the SEM. If you take the signal at the monitor, that includes the signals that are displayed as micron bars, etc. Of course that makes it easier to acquire the signals, but because there is no change of the microscope, there are no additional capabilities. EDS dot maps can be read, but again you depend on another piece of equipment to give you the signals and the correlation with the beam position. In most cases users are interested in getting high quality images, which means that you would run the SEM in photo mode. If your SEM is limited to 1000 lines, the images you get are roughly 1200 x 1000 (determined through the aspect ratio of the screen), or 2400 x 2000 if you have a 2000 line photo mode. Depending on how easy it is to change the photo parameters, it could be simple or hard to change the parameters for the images (for example dwell time, etc.)
With an active system, the digital interface includes a scan generator. While the system is active, it replaces the SEM internal scan generator, but uses all of the other electronics. Aside from the fact that one does not have to replace all the electronics, this also preserves all the settings for astigmatism and other parameters between switching to internal or external. Since the scan generator is now under direct control of the computer, the beam can be moved wherever the computer dictates, and kept at any point as long as the computer decides. This allows the computer total freedom of image resolution and aspect ratio (for example a very rapid frame rate at a reduced resolution for focusing, or a high resolution at longer dwell times for high quality images.) It is also easier to acquire dot maps as the computer knows at any time where exactly the beam is and only needs the number of counts from the EDS system. In terms of image resolution, there is a trade-off: The beam position is determined by the voltage the scan generator produces, modified by the scan amplifier. A normal scan voltage is of the order of a few volts. If this is digitized into 1000 positions, the voltage for 2 neighboring positions differ by a few millivolts. All the electronics adds noise, so if you go to high in resolution, the voltage difference of two neighboring pixels can be of the same order as the noise on the signal. In other words, going too high in resolution could add significant time to the acquisition but not result in better images.
Which one is better? I don't think there is a final answer to this. If your goal is to simply digitize the images on the SEM, and you don't have any issues with the resolution and aspect ratios, etc, a passive system will work just fine. If you need higher resolution, need to control dwell time better, or need other control of the SEM, a passive system will not give you enough possibilities. Or make sure that the passive system is upgradeable to an active system if that becomes necessary at some time.
'nuff said. I wasted too much bandwidth already...
mike
} } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Ave #300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ------------------------------------------------------------------------ } 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 a Philips 515 SEM (with analogue image formation) and I wish to print digital images from it. } What are the advantages/disadvantages of using an active vs. a passive slow scan image grabber to } acquire images? } } }
Where is your response relative to my posting? Do you have a way of deliniating your input from others'?
Otherwise, I cannot sort out your nonsense from all other.
gg
At 07:49 PM 2/24/2002, you wrote:
} -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Sunday, February 24, 2002 3:35 PM } To: Ascend_jcr } Cc: MSA listserver } Subject: Re: FW: Active or Passive Frame Grabber for aqnalogue SEM? } } } I think that an active system does create new capabilities. } It allows different aspect ratios and also variable pixel } dimensions. Why would you want non-square pixels? Philips did this with } their XL system and destroyed any possibility of ever doing image analysis } with the images. } Additionally, unless the SEM allows for } legends to be turned off, (all indeed provide this capability) only any } active system will } capture a pure image (no alpha stuff) so that it can be } image processed later on. Many times, the alpha } creates anomalies during processing. Sometimes it } does not. Without alpha, it is not an issue. It is not an issue on any SEM } } I find that the requisite pixel dwell time is inversely } proportional to the pixel dimensions. i.e., the more } pixels, the shorter the dwell time. With small pixel } dimensions, longer pixel dwell time reduces noise } which also happens with larger pixel dimensions but } at shorter dwell times. This is baloney. The pixel size is related to the } DACs digital to analog conversion. } Either reduce noise at small } pixel dimensions via longer dwell time or increase } pixel dimensions and reduce dwell time. For image } processing, I like 3088x2060 at 5uS. This replaces } the recording CRT's 1.33 aspect ratio to 1.5--which } fits nicely on a 35mm slide. Or, change it to 1.33 } and output to 6x4.5cm slides. With an active system, } there is an option, with passive, not. } Passive has a full choice of matrices. } } The SEM's scan system is of course necessary to } do stig, positioning, etc. Whether the EDS system } comes with beam control or not (of course it will), } unless the SEM is designed to allow external beam } control, one needs schematics and some soldering } to gain access to the scan driver amplifiers. But he } does not mention EDS requirements, just digital capture. } So all of the EDS stuff is irrelevant. If the SEM is } designed properly, its imaging system will be sync'd } to line frequency. Active systems can do this too. } Furthermore, some can add micron markers, other } data and text. It is optional for the operator to do } so or not. } } A passive system is certainly going to be cheaper. } If that is an issue, go for a passive system. My system } is both active and passive. I don't use the passive mode. } Fortunately, my SEM provides slow scan info as RS-170. } This is easily framegrabbed to accompany a high resolution } active scan. Neat. } } BTW, the current Rontec EDS system is active scan. } Does a nice job. } } gary g. } } } } At 10:32 AM 2/24/2002, you wrote: } } } -----Original Message----- } } } From: Ascend_jcr [mailto:ascend_jcr-at-att.net] } } Sent: Sunday, February 24, 2002 10:28 AM } } To: gbarclay-at-trinidad.net; Microscopy-at-sparc5.microscopy.com } } Subject: RE: Active or Passive Frame Grabber for aqnalogue SEM? } } } } } } Greg, } } } } I couldn't help comment on this situation as Mr. Feingold obviously has } some } } hidden agenda or doesn't grasp the concepts fully. First, an active system } } will not create new capabilities. It simply duplicates what your SEM does } } with the addition of independant digital sampling. Yes, it can drive the } } beam, but rarely better than the original SEM hardware which you will still } } need, incidentally for correcting stig, alignments, etc. What he says about } } EDS is partially true but most EDS systems (including older ones) come with } } beam control packages. The advantage of getting the beam control from the } } EDS system is that they provide integrated software to do digital mapping, } } linescans, in addition to electron imaging. Just purchasing an archiving } } system, limits you to electron imaging: you would then have to undertake a } } multi-man-year project to integrate with the EDS spectrometer. } } None of the second part of Feingold's response is accurate. Slower dwell } } times generally do not improve the image for two reasons: First, is that } you } } tend to build up charging and/or contamination on a stationary spot (are } you } } familiar with the dark square left behind by the raster?). Secondly, all } } SEMs have a stage drift factor from signififcant to severe. Sitting on a } } stationary spot will actually result in a record of the stage drift. This } is } } why the EDS companies provide elaborate drift compensation programs with } } their digital beam control packages. } } } } A further disadvantage of an active system is that it loses all instrument } } information unless it maintains a separate communication line with the SEM } } for recording kV, mag, etc. } } } } Passive systems are far superior for pure image collection applicatons such } } as digital image archiving. They retain the microscope information printed } } on the data bar. They can achieve higher resolution and in some cases } } perform frame averaging which is a for superior method of reducing noise } } over point averaging since charge doesn't build up. } } } } I have access to both types systems in addition to a system provided by } Edax } } and would be happy to provide more detailed information if you want. Is } } Feingold trying to sell an active system? I didn't know there were any left } } on the market. } } } } Regards, } } Joe Robinson
Just to add to this, I was asked the same question a few weeks ago, and the combination of FDA, as outlined by Kristen, and propidium iodide (PI), is also very good. PI (at about 1-10 microgram/ml final concentration in water or buffer) penetrates non-viable cells giving red fluorescence and provides a good contrast to the green FDA, especially in pollen that is highly autofluorescent. It can be excited with blue or green light - 488, 514 or 543 nm laser lines on confocal. cheers, Rosemary
} } Hi Soumitra, } Yes, there is a quick and easy way. I don't have my original protocol, but } this one will probably do (from Ruzin's Plant Microtechnique and } Microscopy). Dissolve 1 mg of fluorescein diacetate (FDA) in 1 mL of } acetone for a stock solution that can be stored at -20C. Then add to pollen } grains for a grand total concentration of 1 microgram FDA/mL and incubate } for 5 minutes. Observe under blue light (488 nm). Living cells will } fluoresce yellow/green. My pollen was usually in germination medium at the } time so that is into what I added the FDA. I'll cross-check this with my } old files and let you know if there are any significant differences. Good } luck. } 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 } www.baumlab.org
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
61- 2 6246 5475 or 61- 0402 835 973 rosemary.white-at-csiro.au
take it from someone that has been around em for over 20 years. it is well woth the extra money for the piece of mind that comes from having a service contract with the the compnay you bought your scope from. it may be less expensive to go with a private company like presto, however the costs down the road in parts, if they can be found and in down time, likly to be longer. like i always say stick with what you know. companies like philips and jeol are know quanites and will be around for a long time. just my nickels worth. you know the price of inflation and my experience leads me to charge more... john all typos are the result od a sticky keyboard.
I need to calculate the interaction volume of the SEM electron beam in my specimen. The Monte Carlo simulation packages that I encountered so far only calculates the volume for pure elements. But I sometimes have oxides with densities that are different from that of the pure metals (something like Al2O3). Is there a program that can calculate the interaction volumes for these materials ?
Thanks in advance Willem Erasmus
Willem Erasmus Snr. Scientist, Basic Catalysis Research Sasol Technology Tel : +27 +16 960 - 4211/2772 Fax : +27 +16 960 - 2826 E-mail : willem.erasmus-at-sasol.com PO Box 1, Sasolburg, 1947, Republic of South Africa
[All views expressed are my own and not necessarily that of my employer.]
Another 2 cents worth on digital imaging on the SEM:
MAINS FREQ INTERFERENCE:
SEMs are almost always affected by mains frequency stray field. As a result the scan generators always lock their horizontal signal to the line frequency. This means that the stray field becomes a distortion rather than a blur. When using a small, rapid scan for focusing and astigmatism correction, this distortion will cause the image to undulate, unless the vertical scan signal is also line-locked.
I don't know of any stand alone active digital scan systems that do this. If they exist, it would be important to plug them into a wall socket using the same phase of the 3-phase power (normally present in laboratories) as is used to run the SEM itself.
More to the point, an active system must use some sort of wire to connect the scan/memory computer to the EM electronics. In spite of all the disclaimers about how "This is not problem with modern electronics and grounding techniques" (claims usually made by digital, not analog, guys), I have never seen any such system that does not create ground loops and hence exacerbate the mains-frequency interference problem mentioned above.
You may not notice it at first if you aren't looking for it but it will be there, especially at low voltage.
Verdict: A strong advantage for passive systems, especially those that make serious efforts to avoid ground loops associated with the video-signal wire (Linearized optical couplings?).
"SIGNAL INTEGRATION"
Another big variable in SEM memories often ignored is the matter of sampling. Early frame grabbers often were converted from video frame-stores. These integrated the signal presented to the ADC for about 60-100 ns, the time needed for a video pixel. However, when recording a 30 second scan with 1000 x 1000 pixels, the pixel dwell time is 30 microseconds, 300x longer. If the integration constant on the ADC wasn't changed (and assuming that of the SEM signal amp was fast), the digitally recorded image was mush noisier than one recorded analog from the screen because it represented only 1/300 th of the signal. Manufacturer's soon changed the circuitry to make the integration constant vary with the scan frequency and raster size (i.e., with the pixel dwell time) but this is easier to do if the digital memory is built into the SEM from the start than with an add on.
Moral here is that having a good a method to keep the time-constant of the ADC proportional to the pixel dwell time at the scan speeds of importance to you is just as significant as any other aspect of SEM digital image memory design. -- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU 3D Microscopy of Living Cells Course, June 10 - 20, 2002, UBC, Vancouver Canada Info: http://www.3dcourse.ubc.ca/ Applications due by March 15, 2002
} Now here is another question. } For a vibrating object, reduce magnification obviously won't catch the motion. } But by blanking the beam periodically(as suggested by Gary and } NASA JPL lab), a crisp image of the vibrating object can be resolved. Or by adjusting } the blanking frequency, it can be seen in a slow motion. } Does that mean the instrument's temporal resolution is improved?
I came up with my own answer: NO This is how I understand the "beam blanking method" works: Suppose an object vibrating at 1Hz between A and B, and the scan rate is 0.1Hz(10s/frame), then I can not see the object moving. there is only an expanded image from A to B. Now if I block the beam periodically at 1Hz. In one cycle, suppose the beam is blocked for 0.9s and unblocked for 0.1s, so that we only can "see" the object within that 0.1s. Since it is synchronized, the object will always come back to the same position(e.g. A1) within that 0.1s period. Therefore I will resolve a crispy image at A1. Now if I change the beam blanking frequency slightly to 1.01Hz. After 10 cycle(while one frame is scanned), the object does not come back to the exact position A1, but a close by position A2. Then a second image will be captured. After 100 cycle, I will have 10 positions of the movement, so it appears that I have resolved it in a slow motion.
Unless my understanding above is wrong, there is no temporal resolution improvement in this process. What this does is to take the information in 100 cycle(expands to 100s) and reconstruct into 1 cycle. We can use this reconstructed 1 cycle to represent all the cycles ONLY because we have the prior knowledge that each cycle is absolutely identical. In another word, if there is an anomaly within one cycle of the vibration, we won't know. I believe there are something parallel in respect of spatial resolution.
Then what has been improved? In my opinion: spatial resolution. In the beginning, we actual can resolve the motion by just change the scan rate from 10s/frame to 0.1s/frame. Then I can see 10 position within one true cycle. What not using this rate?
Because it is a "fast scan", we won't see each image clearly. We know we can observe better spatial resolution at "slow scan", the obove "beam blanking method" allows us to obtain 10 clear images. Although the spatial resolution is improved, the true instrument temporal
resolution is REDUCED.
Here is my definition: the temporal resolution IS scan rate, i.e. if the scan rate is 0.1s/frame, the temporal resolution is 0.1s.
Therefore, non-scanning microscopy(LM, TEM) has infinite temporal resolving power and only
limited by their aquisition system(eye, video camera, etc).
Imagine observing a slow flying bee(fast vibrating wings and slow moving body): By using "beam blanking" method in a scanning microscope to get a "slow motion", we see: Slow flapping wings, fast moving body. By using an ultrafast vedio camera in non-scanning miroscope and re-play in slow motion, we see: Slow flapping wings, almost still body.
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Has anyone out there had any success with high pressure freezing/freeze substitution of plant leaves for TEM examination? If so, please contact me as I haven't.
Bob
-- Robert R. Wise, Ph.D. Associate Professor of Plant Physiology Department of Biology and Microbiology University of Wisconsin Oshkosh
On sabbatical from Sept. 2001 to Aug. 2002 at UW Madison Botany Department B217 Birge Hall 430 Lincoln Drive Madison, WI 53706 (608) 262-4288 (phone) (608) 262-7509 (fax) wise-at-uwosh.edu http://www.wisc.edu/biotron/Sharkey/ http://www.uwosh.edu/departments/biology/wise/wise.html
Although rather expensive, I believe a program called Electron Flight Simulator should do what you need.
The web address is: http://www.small-world.net/efs.htm
Woody White McDermott Technology, Inc. ------------------------------------------------------------
} } Dear listmembers } } I need to calculate the interaction volume of the SEM } electron beam in my specimen. The Monte Carlo simulation } packages that I encountered so far only calculates the volume } for pure elements. But I sometimes have oxides with densities } that are different from that of the pure metals (something } like Al2O3). Is there a program that can calculate the } interaction volumes for these materials ? } } Thanks in advance } Willem Erasmus } } Willem Erasmus } Snr. Scientist, Basic Catalysis Research } Sasol Technology } Tel : +27 +16 960 - 4211/2772 } Fax : +27 +16 960 - 2826 } E-mail : willem.erasmus-at-sasol.com } PO Box 1, Sasolburg, 1947, Republic of South Africa } } [All views expressed are my own and not necessarily that of } my employer.] } } }
John, So glad you keep such an open mind. The sticky keyboard is also the fault of third party service, right? Thought so.
DISCLAIMER: I have a vested interest in providing high quality service to users of SEMs. It's how I make an honorable living and take care of my family while helping a lot of wonderful people do THEIR jobs better.
Ken Converse owner Quality Images third party SEM service since 1981 Delta, PA
Soft-Imaging ADDA locks to mains. I suspect that Rontec's imaging system does too. ADDA allows for locking or not. If you don't believe this, I can send you a screen capture of the setup window which offers this user option.
ADDA has a PCI board in the PC but communicates to a separate D/A and A/D box via fiber optic cables. The separate box is electrically at the same potential as the SEM--it knows nothing about the potential at the PC or any difference in potential between the SEM and PC. It is a non-issue.
Furthermore, well-designed active systems use a single wire for ground between the SEM and drive electronics. The analog drive signals and scan generator on/off are conveyed via shielded cables which are only grounded at the driver side-- not at the SEM end. Another move to eliminate ground loops.
Could you expand on the "signal integration" section? I'm not following you, sorry. TV video is 15KHz per line or 67uS. This is not what is being sampled. It is slow scan that is being sampled. How often is based on the number of pixels per line. That can vary.
gary g.
At 07:38 AM 2/25/2002, you wrote:
} Another 2 cents worth on digital imaging on the SEM: } } MAINS FREQ INTERFERENCE: } } SEMs are almost always affected by mains frequency stray field. As a } result the scan generators always lock their horizontal signal to the } line frequency. This means that the stray field becomes a distortion } rather than a blur. When using a small, rapid scan for focusing } and astigmatism correction, this distortion will cause the image to } undulate, unless the vertical scan signal is also line-locked. } } I don't know of any stand alone active digital scan systems that do this. } If they exist, it would be important to plug them into a wall socket using } the same phase of the 3-phase power (normally present in laboratories) as } is used to run the SEM itself. } } More to the point, an active system must use some sort of wire to connect } the scan/memory computer to the EM electronics. In spite of all the } disclaimers about how "This is not problem with modern electronics and } grounding techniques" (claims usually made by digital, not analog, guys), } I have never seen any such system that does not create ground loops and } hence exacerbate the mains-frequency interference problem mentioned above. } } You may not notice it at first if you aren't looking for it but it will be } there, especially at low voltage. } } Verdict: A strong advantage for passive systems, especially those that } make serious efforts to avoid ground loops associated with the } video-signal wire (Linearized optical couplings?). } } "SIGNAL INTEGRATION" } } Another big variable in SEM memories often ignored is the matter of } sampling. Early frame grabbers often were converted from video } frame-stores. These integrated the signal presented to the ADC for about } 60-100 ns, the time needed for a video pixel. However, when recording a 30 } second scan with 1000 x 1000 pixels, the pixel dwell time is 30 } microseconds, 300x longer. If the integration constant on the ADC wasn't } changed (and assuming that of the SEM signal amp was fast), the digitally } recorded image was mush noisier than one recorded analog from the screen } because it represented only 1/300 th of the signal. Manufacturer's soon } changed the circuitry to make the integration constant vary with the scan } frequency and raster size (i.e., with the pixel dwell time) but this is } easier to do if the digital memory is built into the SEM from the start } than with an add on. } } Moral here is that having a good a method to keep the time-constant of the } ADC proportional to the pixel dwell time at the scan speeds of importance } to you is just as significant as any other aspect of SEM digital image } memory design. } -- } ********************************************** } Prof. James B. } Pawley, Ph. 608-263-3147 } Room 223, Zoology Research } Building, FAX 608-265-5315 } 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU } 3D Microscopy of Living Cells Course, June 10 - 20, 2002, UBC, Vancouver } Canada } Info: http://www.3dcourse.ubc.ca/ Applications due by March 15, } 2002
I am looking for an antibody that recognizes the DNA binding domain of Gal4 and that can detect antigens in tissue using indirect immunofluorescence or imunohistochemistry, I would appreciate information! Thanks
I think that a major factor about SEM digitization is being underestimated. If not ignored--regardless of active or passive issue.
Software. The software that accompanies the hardware is perhaps more important and has more impact on results than does the hardware. Simply capturing an image is one thing, but being able to work with the image, filter it, modify it, archive it, index it, etc. are big factors I would think in the grander scheme.
Dear Vitaly, While I don't really want to wade into the various advantages of active vs. passive digital imaging, I would like to correct a misconception presented by several people about EDS imaging. The Quartz XOne EDS system now offers completely passive, full-spectrum mapping and line scans, based on the passive imaging capabilities of the Quartz PCI system. The maps are accumulated over several scans at the line resolution of your SEM and this makes the acquisition of 1024 X 840 x-ray maps in ten to fifteen minutes quite feasible. Also, by integrating and averaging several high-resolution scans, many of the advantages of the longer dwell times available in the active system can be realized by the passive. Disclaimer: I have been involved in the design, testing and selling of the Quartz XOne EDS system since its inception. At 06:59 PM 2/23/2002 -0500, you wrote: } } Greg, } } A quick answer is- advantages of an active system can be divided in two } categories. } } 1) Principal. An active system will create new capabilities, which your SEM } doesn't have now. For example, modern PC or a Mac. computer control of } applications like electron beam lithography and EDX imaging, which depend on } synchronization of either beam blanker or an EDX spectrometer with the } e-beam position. Another principal advantage, is that a large portion of SEM } electronics (most of the scan generation and video signal processing related } circuits) will no longer be needed. This will make an SEM more reliable and } less expensive to maintain. } } 2) Non-principal. An active system will allow much slower scan speed than } the SEM scan generator, and much longer integration times (pixel dwell } times) for video signal acquisition. That will allow you to acquire higher } resolution images easier than with the passive system. Also, active system } can better compensate for AC fields related interference, as compared with } the passive system. } } Active system is more expensive than the passive one. A motorcycle is more } expensive than the bicycle. I wouldn't call it a disadvantage. } } Vitaly Feingold } Scientific Instruments and Applications } 2773 Heath Lane, Duluth GA 30096 } (770)232-7785 ph. } (770)232-1791 fax } (678)467-0012 mobile 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
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Dear William,
Simple Monte Carlo Programs do not account for the distortion of the trajectories caused by charge build-up in insulators.
I actual fact this may be the major factor in determining the size and shape of the interaction volume (not to mention the shape of the probe that actually reached the surface of an uncoated insulator)
Experiments in this field often go back to EDX studies of frozen aqueous solutions in which the tail of the Bremsstralung consistently failed to reach the beam voltage and the beam was later shown not to have "penetrated" as far as it "should" for this Z and density.
As charging itself is extremely complex, I would state it as my opinion that Monte Carlo simulations of beam penetration in insulators are of marginal utility unless you are only interested in the first few microseconds of beam residence time on a surface known not to have any trapped charge to start with.
Cheers,
Jim P.
-- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU 3D Microscopy of Living Cells Course, June 10 - 20, 2002, UBC, Vancouver Canada Info: http://www.3dcourse.ubc.ca/ Applications due by March 15, 2002
{color} {param} 0100,0100,0100 {/param} {FontFamily} {param} Times New Roman {/param} {bigger} Greetings,
I needed a company to move a Joel SEM with EDX from central New Jersey to Cleveland Ohio.
The Cleveland Museum of Natural History has had an offer to have a Joel SEM donated. The catch is that it is in the basement of a consultants house. Does anyone know of a company in the Ohio-Pennsylvania-New Jersey (New York?) area who is, at the least, capable of hoisting it out of his basement and onto a truck? We may also be interested in delivery and re-assembly and alignment.
Thank you for any suggestions,
Dr. David Saja, Geologist
dsaja-at-cmnh.org {/color} {FontFamily} {param} Courier New {/param} {smaller}
{nofill} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. David Saja (216) 231-4600 x429 Curator of Mineralogy FAX: (216) 231-5919 dsaja-at-cmnh.org
The Cleveland Museum of Natural History 1 Wade Oval Drive, University Circle Cleveland, Ohio 44106-1767 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Good points (and definitely worth more than 2 cents!).
Mains Frequency Interference: Yes, this is quite obvious sometimes. For that reason our system does include the synch electronics. You pay a small price in terms of speed. Since each line has to be synched to a certain phase of the power signal, each line is delayed a bit, leaving to a slighly lower frame rate. I can only speak for our electronics, but I am sure others have that as well.
Ground loops: Yes, they can be a problem. That is why we try to keep the copper wires as short as possible by placing a box somewhere near the SEM, and then connect the box to the PC via an optical connection. Of course the optical connection is immune to electro-magnetic interference, but you're right, if you are not careful, even the shortest cables can suffer from interference. And if you look in the back of your SEM, there are plenty of cables that could interfere! Sometimes, at the highest magnifications, mechanical vibrations may be confused with electrical interference.
Signal Integration: I would consider this an advantage of the active system, as the computer can determine the dwell time as opposed to a passive system where the SEM determines the dwell time and the computer has to "guess". But you are of course correct, in that it is not sufficient to simply use a 60 ns integration time. However, keep in mind, that the X-direction in older SEMs is NOT digitized. It is an analog signal. There is no real "dwell time" as the beam is in constant movement. The "x-resolution" is then determined by the number of lines, the requirement to have square pixels, the aspect ratio of the screen, and of course the time for one line scan. For example, if you use a 1000 line photo setting, the final picture should have something like 1280 x 1000 pixels (The aspect ratio of the SEM screen is usually 4:3). For a total frame time of 2 min (120 sec), the line time is roughly 120 msec (120/1000), resulting in a "pixel itme" of roughly 100 microseconds (120 msec / 1280).
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: James Pawley [mailto:jbpawley-at-facstaff.wisc.edu] Sent: Monday, February 25, 2002 8:38 AM To: Microscopy-at-sparc5.microscopy.com
Another 2 cents worth on digital imaging on the SEM:
MAINS FREQ INTERFERENCE:
SEMs are almost always affected by mains frequency stray field. As a result the scan generators always lock their horizontal signal to the line frequency. This means that the stray field becomes a distortion rather than a blur. When using a small, rapid scan for focusing and astigmatism correction, this distortion will cause the image to undulate, unless the vertical scan signal is also line-locked.
I don't know of any stand alone active digital scan systems that do this. If they exist, it would be important to plug them into a wall socket using the same phase of the 3-phase power (normally present in laboratories) as is used to run the SEM itself.
More to the point, an active system must use some sort of wire to connect the scan/memory computer to the EM electronics. In spite of all the disclaimers about how "This is not problem with modern electronics and grounding techniques" (claims usually made by digital, not analog, guys), I have never seen any such system that does not create ground loops and hence exacerbate the mains-frequency interference problem mentioned above.
You may not notice it at first if you aren't looking for it but it will be there, especially at low voltage.
Verdict: A strong advantage for passive systems, especially those that make serious efforts to avoid ground loops associated with the video-signal wire (Linearized optical couplings?).
"SIGNAL INTEGRATION"
Another big variable in SEM memories often ignored is the matter of sampling. Early frame grabbers often were converted from video frame-stores. These integrated the signal presented to the ADC for about 60-100 ns, the time needed for a video pixel. However, when recording a 30 second scan with 1000 x 1000 pixels, the pixel dwell time is 30 microseconds, 300x longer. If the integration constant on the ADC wasn't changed (and assuming that of the SEM signal amp was fast), the digitally recorded image was mush noisier than one recorded analog from the screen because it represented only 1/300 th of the signal. Manufacturer's soon changed the circuitry to make the integration constant vary with the scan frequency and raster size (i.e., with the pixel dwell time) but this is easier to do if the digital memory is built into the SEM from the start than with an add on.
Moral here is that having a good a method to keep the time-constant of the ADC proportional to the pixel dwell time at the scan speeds of importance to you is just as significant as any other aspect of SEM digital image memory design. -- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU 3D Microscopy of Living Cells Course, June 10 - 20, 2002, UBC, Vancouver Canada Info: http://www.3dcourse.ubc.ca/ Applications due by March 15, 2002
} Soft-Imaging ADDA locks to mains. I suspect that } Rontec's imaging system does too. ADDA allows } for locking or not. If you don't believe this, I can } send you a screen capture of the setup window } which offers this user option.
I have no reason not to believe you. Many early systems lacked this facility. I am glad that your does not.
} ADDA has a PCI board in the PC but communicates } to a separate D/A and A/D box via fiber optic cables. } The separate box is electrically at the same potential } as the SEM--it knows nothing about the potential at } the PC or any difference in potential between the } SEM and PC. It is a non-issue.
I am glad that you use F/O couplings but, with respect, whether or not it is "a non-issue" would be determined when I hooked your system attached to my Hitachi S-900 FESEM working at 1kV, and found there was no distortion at 200kx. I know of no more sensitive ground-loop sensor.
I am not saying that it cannot be done, just that it seldom is done and people are often not even aware that it can be a problem.
} Furthermore, well-designed active systems use } a single wire for ground between the SEM and } drive electronics. The analog drive signals and } scan generator on/off are conveyed via shielded } cables which are only grounded at the driver side-- } not at the SEM end. Another move to eliminate } ground loops.
This all sounds fine but one usually adds a frame store to an old scope (new ones have their own). AFTER they have been installed in your lab such scopes often have grounding systems that have surprising properties.
} Could you expand on the "signal integration" section? } I'm not following you, sorry. TV video is 15KHz } per line or 67uS. This is not what is being sampled. } It is slow scan that is being sampled. How often } is based on the number of pixels per line. That can } vary.
I merely wish to point out that while photographic image collection automatically integrates the signal for each pixel in the time domain (even if it weren't already "time-averaged" by the slow response of the CRT phosphor), this is not automatically the case for digital frame stores.
Unless the system employs "box-car" integration (which is a great idea! It is found on Bio-Rad confocal scopes) the bandwidth of the preamp before the ADC determines an integration time. If this time is less than the time that the SEM beam spends traversing one pixel in the raster, the voltage sensed by the ADC will be averaged only over part of the pixel time and the resulting data will be noisier than it should be.
The problem is more important in the SEM than in other digitally sampled imaging systems because the pixel-dwell time can vary widely: from 0.1 microseconds at "tv-rate" to 1000x that for slow scan. The signal preamp bandwidth on the scope must be wide enough to pass the high-frequency "tv-rate" signal. But hat means that it is much too wide to properly integrate the slow scan signal. The bandwidth of the signal being fed to the ADC must be adjusted in the frame-store to correspond to the pixel dwell time.
We won't go into how the PSF of the microscope itself limits the bandwidth of the signal. (At low mag very large changes in signal level from one pixel to the next are possible: at high mag, things get fuzzy and large changes aren't possible.) But in principle, it would be possible to use magnification and other information from the SEM to impose additional limits on the bandwidth of the ADC. As I remember, Everhardt proposed changing the bandwidth of the video amp with the signal level and the scan speed in his Thesis in the 1950s.
Alternatively, John MacKensie recommends that when sampling "slow" (long?) pixels, you use the same fast ADC-preamp settings but make many measurements during the long pixel dwell time and then digitally average the results before sending the value off to storage. Assuming that this super-sampling rate was similar to the time constant of the ADC preamp, this should work well.
This is not a small problem. Some years ago David Joy recorded white noise images by having a defocused beam strike smooth specimen without scanning. The only variations in the SE signal should have been the statistical variations in the SE current caused by shot noise. As he could measure this current, he could calculated what shot-noise should be and then calculate a average-signal to shot-noise ratio. He could also measure the standard deviation of the image data stored in the memory and ratio this to the average intensity.
If the detector had high quantum efficiency and the digitizer worked properly, these two S/N ratios should be the same. (i.e., the data recorded would have the same S?N ratio as the signal leaving the specimen.)
In fact, on all of a large number of different model instruments, the digitally recorded signal was noisier than it should have been (the best was 2x worse, the worst almost 1000x worse!!). Clearly some particular microscope models were much worse than others. A factor of 10 converts to having to use either 10x more beam current or 10x longer scan time to record the same image as would have been possible in analogue/photopraphic mode.
At the time of this publication, it wasn't clear how much of the discrepancy was due to poor design of the SE detector and how much due to differences in digitization circuitry.
My own opinion is that digitization was the cause of most of the instrument-to-instrument variation and poor detector design was the reason that none of the instruments really came close to theoretical performance.
These results were published in Scanning Volume 18-8, November (1996) Measuring the Performance of Scanning Electron Microscope Detectors D.C. Joy, C.S. Joy, R.D. Bunn pp 533.
It is a fascinating paper that shows how to measure a reproduceable and significant difference between instruments that is at least as important as any of the "Specs" that they are usually sold on the basis of.
We can hope that, in the intervening time, manufacturers have responded to the problem and that if the test were carried out today, the differences would be less. Or alternatively, we can follow Joy protocol and test it for ourselves.
Cheers,
Jim P. -- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU 3D Microscopy of Living Cells Course, June 10 - 20, 2002, UBC, Vancouver Canada Info: http://www.3dcourse.ubc.ca/ Applications due by March 15, 2002
The program Electron Flight Simulator can do the job for you. You can put any elements together and calculate the density of the mixture and program will solve the simulation. It can work on simulation of bulk, film layers on substrate, particles on top and variable presure simulation. In my SEM lab, it is tied to ISIS-300 EDS system (I do not know if you can run it on regular computer). I have no personal interest with Electron Flight Simulator.
Thanks Zhiyu Wang Maxtor Corp.
----- Original Message ----- } From: "White, Woody N." {nwwhite-at-mcdermott.com} To: "'Erasmus, Willem (WJ)'" {willem.erasmus-at-sasol.com} ; {Microscopy-at-sparc5.microscopy.com} Sent: Monday, February 25, 2002 6:00 PM
Dear All
We are attempting view Tardigrades in a fei/Phillips e-sem. We were unable to view them in the active state. They curl up in the microscope. When the oxygen is removed from their environment they become non-active but bloated. This is what we want. Since they have a soft exoskeleton they also collapse easy. Probably use the internal water pressure to maintain their body volume. Has anyone out there had any success with these magnificent beasts? If so, please HELP!
Mr. S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gabarone Botswana
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I have recently read some messages about the Nyquist sampling rate. To know the appropriate sampling rate for a given CCD-camera on a microscope I have made a table in which the N.A. of an objective lens can be related to the necessary magnification needed to sample the image on a CCD-grid to satisfy the Nyquist sampling rate.
Although sampling at the Nyquist rate will give you nice images, it is not yet sufficient to give you "reliable" quantitative results in mesaurements! The Coefficient of Variation (C.V.) in your measurements decreases with the increase of the diameter of the object as it is projected on your CCD-array. There are a few very nice publications from Prof. Ted Young (T.U.Delft, the Netherlands) on this subject. As a general rule I take that an object (diameter) should cover at least 20 pixels on your CCD-grid to bring the C.V. down to a reasonable level.
Some references:
Ian T. Young, Not just pretty pictures: Digital quantitative microscopy, Proc. Royal Microscopical Society, 1996, 31(4), pp. 311-313.
Ian T. Young, Quantitative Microscopy, IEEE Engineering in Medicine and Biology, 1996, 15(1), pp. 59-66.
Ian T. Young, Sampling density and quantitative microsocopy Analytical and Quantitative Cytology and Histology, vol. 10, 1988, pp. 269-275
These rules are applicable to 2D microscopy, but they can be extended to confocal microscopy. The difference in sampling in the Z-dimension on a confocal microscope will probably relate different to this appropriate sampling density compared to the XY-dimension.
Best regards,
Dr. Peter Van Osta
Union Biometrica N.V./S.A. European Scientific Operations
Dear Listmembers, I would like to know how to soften hardwood for slide preparation. I would like to share the techniques of softening, sectioning (sliding microtome), staining and mounting. Anyone want to share please email me.
Regards Hasidin A. R. Wood Anatomy Lab Faculty of Forestry Universiti Putra Malaysia 43400 Serdang Selangor Malaysia.
We do not use such a program here. My only experience (albeit very limited) is with two programs: Aldos "Fetch" and "Quartz PCI". Gatan promotes (promoted?) Fetch. We have at least one copy. Quartz PCI, the 3rd party imaging and image manipulation software marketed by Hitachi, contains an archiving software.
My opinion is that any archiving software requires more up-front work than it is worth. We choose, for the most part, to keep good records on the sample types pertaining to each work request and locate images based on this information. Thus, we tend to spend our time ensuring that we can locate our archived files. The LabLan has become very useful toward this purpose.
Another source to check is the microscopy list server www.msa.microscopy.com/MicroscopyListserver. One can search their discussion threads for information on a variety of topics. One should note that, for the most part, the comments on the list server are opinion. Remember that opinions are like noses: everyone has one and my having one is perfectly obvious to everyone else.
Good luck and give my best to Johan and the rest of the MCM gang.
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Alistair D Westwood To: Gary M Brown/Baytown/ExxonMobil-at-xom, Jerry W Ball/Baytown/ExxonMobil-at-xom, Sandra M Wapp/Baytown/ExxonMobil-at-xom, David W 02/26/02 06:55 Abmayr/Baytown/ExxonMobil-at-xom AM cc: Subject: image management software
Folks,
Any comments on Anton-Jan's email.
Ali
Alistair D. Westwood Team Leader - Microscopy & Surface Science Materials Characterization Lab - Polymer Science Division ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, TX 77520
Ph: (281) 834-5741 Fax: (281) 834-1793 ----- Forwarded by Alistair D Westwood/Baytown/ExxonMobil on 02/26/02 06:54 AM -----
Anton-Jan Bons To: Alistair D Westwood/Baytown/ExxonMobil-at-xom, Mark M 02/26/02 01:14 Disko/East-US/ExxonMobil-at-xom AM cc: Johan Stuyver/Benelux/ExxonMobil-at-xom, Marc H Anthonis/Benelux/ExxonMobil-at-xom Subject: image management software
Mark, Ali, We are looking for a software package to manage our microscopy images. We have PhotoShop, but that's not ideal for batch file conversions, browsing through large numbers of images, etc. We have a demo version of LView Pro ( http://www.lview.com/AboutLViewPro.htm) which looks very convenient; it's only 50$ but of course we canot order it just like that... Do you use software for image management? Do you have any suggestions? It would be best if we all use the same software.
Thanks. Regards, Anton-Jan Bons ExxonMobil Chemical - European Technology Center Hermeslaan 2, B-1831 Machelen, Belgium tel: +32 2 722 2838, fax: +32 2 722 2461
Morning Bob, I hope this is on point, but as a zoologist, I would like to know how the mesophyl (?-no elementary bio book to check on leaves anymore) holds up at 30K psi? You may have the same problem with a leaf as I might have with a piece of mammalian lung. At 30K psi I can only imagine that the piece of lung would simultaneously be frozen and compressed - to me, that means, destroyed. Most of this has been imagined, so I won't spend any more time on it. Interestingly, the only botany book I have at the moment in my library is by Johansen, D.A, Plant Embryology, 1950 (Cycads to Anthophyta???- Spermatophyta (of the time?)), and, of course, no leaves in it.
Regards,
Fred Monson
} ---------- } From: Bob Wise } Sent: Monday, February 25, 2002 12:39 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: HPF of leaf tissue } } ------------------------------------------------------------------------ } 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 out there had any success with high pressure } freezing/freeze substitution of plant leaves for TEM examination? If } so, please contact me as I haven't. } } Bob } } -- } Robert R. Wise, Ph.D. } Associate Professor of Plant Physiology } Department of Biology and Microbiology } University of Wisconsin Oshkosh } } On sabbatical from Sept. 2001 to Aug. 2002 at UW Madison } Botany Department } B217 Birge Hall } 430 Lincoln Drive } Madison, WI 53706 } (608) 262-4288 (phone) } (608) 262-7509 (fax) } wise-at-uwosh.edu } http://www.wisc.edu/biotron/Sharkey/ } http://www.uwosh.edu/departments/biology/wise/wise.html } }
Dear David, You might have some luck contacting a piano moving company, if you don't have a company that speciallizes in moving hi-tech equipment. The piano movers are used to getting heavy, delicate instruments out of awkward places. At 04:25 PM 2/25/2002 -0500, you 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 -----------------------------------------------------------------------. } } Greetings, } } I needed a company to move a Joel SEM with EDX from central New Jersey to Cleveland Ohio. } } The Cleveland Museum of Natural History has had an offer to have a Joel SEM donated. The catch is that it is in the basement of a consultants house. Does anyone know of a company in the Ohio-Pennsylvania-New Jersey (New York?) area who is, at the least, capable of hoisting it out of his basement and onto a truck? We may also be interested in delivery and re-assembly and alignment. } } Thank you for any suggestions, } Dr. David Saja, Geologist } dsaja-at-cmnh.org 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
Dear Mr. Hasidin, The time I prepared mahogany for SEM examination, I softened it in hot or boiling water for a while (half an hour?), then sliced it with a razor blade. At 04:50 PM 2/26/2002 +0800, you wrote: } } Dear Listmembers, } I would like to know how to soften hardwood for slide preparation. I would } like to share the techniques of softening, sectioning (sliding microtome), } staining and mounting. Anyone want to share please email me. } } Regards } Hasidin A. R. } Wood Anatomy Lab } Faculty of Forestry } Universiti Putra Malaysia } 43400 Serdang Selangor } Malaysia. } 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
What was supposed to be a simple question has produced discussion that goes well beyond what I had expected or what I have the expertise to be able to deal with. I am passing some of the postings on to my colleagues and we are chewing on everything as we try to decide what to do with our old SEM. At least it is essentially new, having sat in a lab for 16 years while politics kept it in mothballs. The great expense for us to upgrade from the Polaroid camera to digital means that we have to be careful what we do.
We are going to purchase a digital capture system for LM and dissection microscope in our histolab. We want to hook it up to a Mac if possible. We don't want to spend a million dollars but we do want something with good resolving abilities and user friendly, is as always, a big plus.
We been having problems getting through to vendors.
Thanks again, Tim Quinn University of Kansas Research Assistant Natural History Museum and Biodiversity Research Center Dyche Hall Room 414 Lawrence, KS 6604-2454 785-864-4556 tquinn-at-ku.edu
Various components of embedding resins present a lab safety problem; many are allergenic and some are potentially carcinogenic. Gloves are the usual precaution, but there's a lot of variation in the protection that they provide, and using the wrong kind can give a very false sense of security. I posted this on the list some time ago:
} Tobler & Freiburghaus recommend bulky, clumsy, expensive "4H" gloves for } } methacrylates [J. Microscopy 160:291-298(1990)], with latex in 2nd place } & } vinyl 3rd. Ringo, Read, & Cota-Robles [J.E.M. Technique } 1:417-418(1984)] found } clumsy, cheap polyethylene much better than latex } in resistance to epoxy } monomers, with vinyl again a poor 3rd. I've heard } comments that "nitrile is } good", but I haven't found any data yet.
Several listers suggested various nitrile glove manufacturers; I contacted them, but none could provide info for the monomers that we use. The CDC has a useful glove chart at http://www.cdc.gov/od/ohs/manual/pprotect.htm. But all that's listed there is methyl methacrylate (nitrile is poorer than latex). Does anyone have any data? Please don't post "just don't spill" comments; safety officers aren't impressed with that attitude.
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
I believe the Optronics Magnafire and Magnafire SP digital cameras work on Mac as well as PC. They were demonstrated here a couple of weeks ago, and both were capable of getting great images on both a compound scope and a dissecting scope. In fact, histo slides on the Olympus stereo scope looked fantastic! The software for the Magnafire SP was easy and intuitive. Contact Optronics or find a distributor on the Web. Olympus is also a distributor, I think.
} We are going to purchase a digital capture system for LM and dissection } microscope in our histolab. We want to hook it up to a Mac if possible. We } don't want to spend a million dollars but we do want something with good } resolving abilities and user friendly, is as always, a big plus.
Aloha, Tina
**************************************************************************** * 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* ****************************************************************************
You can try to call JEOL directly. Perhaps they will do it for you or could make recommendations. I only have their central New Jersey service dept. phone number...(732) 254-5220. Hopefully they will be able to help.
Also, "Allied Van Lines, Inc." has a Special Products Division called "Electronics Van Inc.". They moved our TEM scope from Massachusetts to NJ. Their number is (408) 615-1880. Allied's main number is (630) 717-3000. Allied will not install the scope. You will still need to contact JEOL or another SEM service company for the installation.
ken, there really isn't any need to be so defensive. i offered my openion on the subject based on over 20 years in the field. your is biased around a vested need to keep you and your family fed. which i find no fault in. i have seen the results of to many companies trying to provide service on instruments they shouldn't even be attempting. as for the keyboard, it was a joke. i think most of those that read it got it. well with one exception. actually it was a laptop with the keys a bit to close together for my rather large fingers.
------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } John, } So glad you keep such an open mind. The sticky keyboard is also the } fault of third party service, right? Thought so. } } DISCLAIMER: I have a vested interest in providing high quality service } to users of SEMs. It's how I make an honorable living and take care of } my family while helping a lot of wonderful people do THEIR jobs better. } } Ken Converse } owner } Quality Images } third party SEM service since 1981 } Delta, PA } } "JHoffpa464-at-aol.com"-at-sparc5.microscopy.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 } } -----------------------------------------------------------------------. } } } } } } take it from someone that has been around em for over 20 years. it is well woth } the extra money for the piece of mind that comes from having a service contract } with the the compnay you bought your scope from. it may be less expensive to go } with a private company like presto, however the costs down the road in parts, } if they can be found and in down time, likly to be longer. like i always say sti } ck with what you know. companies like philips and jeol are know quanites and wil } l be around for a long time. } } just my nickels worth. you know the price of inflation and my experience leads } me to charge more... } } john } } all typos are the result od a sticky keyboard. } } } } } } } }
I didn't expect of my quick comment to generate such a feedback. So, I took a liberty to comment more on this subject. My apologies to the list, shall anybody find this boring.
} Yes, it can drive the } beam, but rarely better than the original SEM hardware which you will } still } need, incidentally for correcting stig, alignments, etc.
Only modern SEM can compete with the PC or MAC controlled interface in driving the beam "better". Modern SEM is an unlikely candidate for such upgrade- all this is about practicality. Further, scan circuits and scan coils of the SEM do not participate in the beam stigmation. This part is irrelevant to the subject. Same applies to beam alignment with respect to the center of the optical column. Alignment, focus, and stigmation will be done in exactly the same way as they were done before the active or passive system installation. Scan rotation and tilt correction (for the most time), are the only circuits relevant to the scan, which will still participate in the process (outside of alignment and stigmation), after the active system is installed. These circuits also do not control SEM scan generator.
} What he says about } EDS is partially true but most EDS systems (including older ones) come } with } beam control packages. The advantage of getting the beam control from the } EDS system is that they provide integrated software to do digital mapping, } linescans, in addition to electron imaging.
EDS system with the beam control package is an active system. With all advantages of such system. So simple. And again, about practicality- older EDS mapping system requires multiple connections with SEM, and complex hardware. Modern active system is all software, both image and EDS spectra/maps, with simple PCI card and interface box. Much more reliable.
} None of the second part of Feingold's response is accurate. Slower dwell } times generally do not improve the image for two reasons: First, is that } you } tend to build up charging and/or contamination on a stationary spot (are } you } familiar with the dark square left behind by the raster?).
Slow scan with high beam dwell time helps to record high resolution images. That hidden agenda calls for the use of a smaller spot size. That means low beam current, low signal, high noise, and high integration time. Low beam current makes for slower charge/contamination grow. Why do a slow scan with high beam current?
} Secondly, all } SEMs have a stage drift factor from signififcant to severe. Sitting on a } stationary spot will actually result in a record of the stage drift. This } is } why the EDS companies provide elaborate drift compensation programs with } their digital beam control packages.
Contamination and stage drift must be repaired, or at least reduced, if they interfere with your application. Otherwise don't worry. SEM users and manufacturers may disagree with the statement (outside the spelling) "all SEMs have a stage drift factor from signififcant to severe", I certainly do. Many SEMs have stage lock for critical applications.
} Is Feingold trying to sell an active system? I didn't know there were any } left on the market.
We are not selling active nor passive systems, however I install and service both, as well as various SEMs/TEMs. Vendors of the active systems, just of the top of my head: http://www.emispec.com/Main/index.html , www.ixrfsystems.com , http://www.4pi.com/ . I am sure there are others- look at http://www.amc.anl.gov/Docs/NonANL/ComSites.html#Instruments .
In order to keep balance in Nature- vendors of the passive systems: http://www.gwelectronics.com/ , www.2spi.com , http://www.orionmicroscopy.com/ . There a others too. As always, MSA web site has many useful links.
Noise generated by the digital signal processing is the separate issue. Prof. James Pawley addressed that - excellent posting. That noise will be generated by both active and passive systems. (The following may seem off subject, but I couldn't help brining this example.) This is one of the reasons why class A vacuum tube amplifier (potentially capable of) reproducing better quality sound, than modern digital audio system. But practicality dictates digital processing, in most instances. Just think about skills and time (number of attempts) required in order to record best possible image by a CRT/Polaroid combination, as compared with PC based system (especially for students). And then scanning the negatives, etc. My initial comment was about practical - off-the-shelf - solution. Passive system is economical, active system has more capabilities. Image quality will be similar in both systems, unless operator pushes SEM to the limit. When he does, extra capabilities of an active system may come handy.
CRT/Polaroid film recording is certainly a bottleneck of image resolution. Different vendors addressed that in different ways in the past. Electromagnetic focus CRT (ISI) is better than electrostatic one. Rodenstock lenses (Cambridge Stereoscan) is better than Polaroid lenses. And so on.
To Mary Mager- thank you for the correction. I was not aware of EDS mapping capabilities of the passive Quartz PCI system- can you post a link?
Cheers.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
This message is made of 100% recycled electrons. ----- Original Message ----- } From: Ascend_jcr {ascend_jcr-at-att.net} To: Net Gbarclay-at-Trinidad. {gbarclay-at-trinidad.net} Cc: microscopy. com Microscopy-at-sparc5. {Microscopy-at-sparc5.microscopy.com} Sent: Sunday, February 24, 2002 1:32 PM
Gary,
you get from an image database what you put in. If you just use it as a replacement of the normal file structure, there is probably more pain than gain. On the other hand, if you sit down and put some effort into it at the beginning, like defining what the keyfields are you want to use for later searches, how the keyfields are supposed to be used, what information you can in automatically or what information you want to "force" the user to put in, it can become a very powerful tool indeed.
Our analySIS software contains an embedded database, and we have users who produce 10s of thousands of images each year and they are happily using the database. In addition, this opens the possibility to allow other people to search and download images through the internet. We not only keep image data in our database but other data as well (EDS data, EELS data, or Word files, Excel, etc...) so you can also use it as a project database.
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: "gary.m.brown-at-exxonmobil.com"-at-sparc5.microscopy.com [mailto:"gary.m.brown-at-exxonmobil.com"-at-sparc5.microscopy.com] Sent: Tuesday, February 26, 2002 7:56 AM To: alistair.d.westwood-at-exxonmobil.com Cc: david.w.abmayr-at-exxonmobil.com; jerry.w.ball-at-exxonmobil.com; sandra.m.wapp-at-exxonmobil.com
Anton-Jan,
We do not use such a program here. My only experience (albeit very limited) is with two programs: Aldos "Fetch" and "Quartz PCI". Gatan promotes (promoted?) Fetch. We have at least one copy. Quartz PCI, the 3rd party imaging and image manipulation software marketed by Hitachi, contains an archiving software.
My opinion is that any archiving software requires more up-front work than it is worth. We choose, for the most part, to keep good records on the sample types pertaining to each work request and locate images based on this information. Thus, we tend to spend our time ensuring that we can locate our archived files. The LabLan has become very useful toward this purpose.
Another source to check is the microscopy list server www.msa.microscopy.com/MicroscopyListserver. One can search their discussion threads for information on a variety of topics. One should note that, for the most part, the comments on the list server are opinion. Remember that opinions are like noses: everyone has one and my having one is perfectly obvious to everyone else.
Good luck and give my best to Johan and the rest of the MCM gang.
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Alistair D
Westwood To: Gary M Brown/Baytown/ExxonMobil-at-xom, Jerry W Ball/Baytown/ExxonMobil-at-xom, Sandra M Wapp/Baytown/ExxonMobil-at-xom, David W 02/26/02 06:55 Abmayr/Baytown/ExxonMobil-at-xom
AM cc:
Subject: image management software
Folks,
Any comments on Anton-Jan's email.
Ali
Alistair D. Westwood Team Leader - Microscopy & Surface Science Materials Characterization Lab - Polymer Science Division ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, TX 77520
Ph: (281) 834-5741 Fax: (281) 834-1793 ----- Forwarded by Alistair D Westwood/Baytown/ExxonMobil on 02/26/02 06:54 AM -----
Anton-Jan Bons
To: Alistair D
Westwood/Baytown/ExxonMobil-at-xom, Mark M 02/26/02 01:14 Disko/East-US/ExxonMobil-at-xom
AM cc: Johan Stuyver/Benelux/ExxonMobil-at-xom, Marc H Anthonis/Benelux/ExxonMobil-at-xom
Subject: image management software
Mark, Ali, We are looking for a software package to manage our microscopy images. We have PhotoShop, but that's not ideal for batch file conversions, browsing through large numbers of images, etc. We have a demo version of LView Pro ( http://www.lview.com/AboutLViewPro.htm) which looks very convenient; it's only 50$ but of course we canot order it just like that... Do you use software for image management? Do you have any suggestions? It would be best if we all use the same software.
Thanks. Regards, Anton-Jan Bons ExxonMobil Chemical - European Technology Center Hermeslaan 2, B-1831 Machelen, Belgium tel: +32 2 722 2838, fax: +32 2 722 2461
I'm getting very tired of those "promotional-type" answers (this listserver had, if I remember well, some rules against this).
We also at HITACHI have a very powerful Digital Image Management System (including very sophisticated Database) called PCI (by Quartz Imaging) but we are NOT using this listserver to promote it
} } } Anton-Jan, } } We do not use such a program here. My only experience (albeit very limited) } is with two programs: Aldos "Fetch" and "Quartz PCI". Gatan promotes } (promoted?) Fetch. We have at least one copy. Quartz PCI, the 3rd party } imaging and image manipulation software marketed by Hitachi, contains an } archiving software. } } My opinion is that any archiving software requires more up-front work than } it is worth. We choose, for the most part, to keep good records on the } sample types pertaining to each work request and locate images based on } this information. Thus, we tend to spend our time ensuring that we can } locate our archived files. The LabLan has become very useful toward this } purpose. } } Another source to check is the microscopy list server } www.msa.microscopy.com/MicroscopyListserver. One can search their } discussion threads for information on a variety of topics. One should note } that, for the most part, the comments on the list server are opinion. } Remember that opinions are like noses: everyone has one and my having one } is perfectly obvious to everyone else. } } Good luck and give my best to Johan and the rest of the MCM gang. } } Gary M. Brown } ExxonMobil Chemical Company } Baytown Polymers Center } 5200 Bayway Drive } Baytown, Texas 77520-2101 } phone: (281) 834-2387 } fax: (281) 834-2395 } e-mail: Gary.M.Brown-at-ExxonMobil.com } } } } } Alistair D } } Westwood To: Gary M } Brown/Baytown/ExxonMobil-at-xom, Jerry } W Ball/Baytown/ExxonMobil-at-xom, } Sandra M } Wapp/Baytown/ExxonMobil-at-xom, } David W } 02/26/02 06:55 Abmayr/Baytown/ExxonMobil-at-xom } } AM cc: } } Subject: image management } software } } } } } } Folks, } } Any comments on Anton-Jan's email. } } Ali } } Alistair D. Westwood } Team Leader - Microscopy & Surface Science } Materials Characterization Lab - Polymer Science Division } ExxonMobil Chemical Company } Baytown Polymers Center } 5200 Bayway Drive } Baytown, TX 77520 } } Ph: (281) 834-5741 } Fax: (281) 834-1793 } ----- Forwarded by Alistair D Westwood/Baytown/ExxonMobil on 02/26/02 06:54 } AM ----- } } } Anton-Jan Bons } } To: Alistair D } } Westwood/Baytown/ExxonMobil-at-xom, } Mark M } 02/26/02 01:14 Disko/East-US/ExxonMobil-at-xom } } AM cc: Johan } Stuyver/Benelux/ExxonMobil-at-xom, Marc } H Anthonis/Benelux/ExxonMobil-at-xom } } Subject: image management } software } } } } } } Mark, Ali, } We are looking for a software package to manage our microscopy images. We } have PhotoShop, but that's not ideal for batch file conversions, browsing } through large numbers of images, etc. We have a demo version of LView Pro ( } http://www.lview.com/AboutLViewPro.htm) which looks very convenient; it's
} only 50$ but of course we canot order it just like that... Do you use } software for image management? Do you have any suggestions? It would be } best if we all use the same software. } } Thanks. Regards, } Anton-Jan Bons } ExxonMobil Chemical - European Technology Center } Hermeslaan 2, B-1831 Machelen, Belgium } tel: +32 2 722 2838, fax: +32 2 722 2461 }
Dear Listers, A protocol for in-well PCR on FAA fixed arabidopsis buds includes "microtoming" a block of the infloresence in 5% agarose. I've already tried a paraffin microtome on a cooled sample and a cryostat but am unable to obtain sections of consistent thickness (40 um). Any ideas or advice is most welcome. Rosemary -- Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
His experience eith other service providers is unusual, our contracts include all spare parts and the response time is mostly faster than the manufacturers. Peter Stolzenberg, Pesto Inc.
I read your response and then read the response that prompted it and I don't think that the vendor went over the line. I thought that the first paragraph was a good response with respect to the benefits of using a database structure. It was certainly within the scope of the topic.
His second paragraph identified it as a product that they are selling (within the rules of the Listserver) and told of the integration that their software had with other data that comes from a microscope such as EDS and EELS spectra. That is certainly within the scope of today's trend in integrating data from an instrument. One only needs to talk to the reps of all of the microscope manufacturers as well as third party manufacturers of Imaging/EDS digital systems. The remarks that he made with respect to the advantages of their software addresses points in the discussion. In addition, his response favored the use of a database management system and did not tout theirs as better than yours. In fact, if one knows the capabilities of PCI as was demonstrated at the Microscopy shows in the Hitachi booths, his arguments do not at all distract from that product.
Let me state that I am not a user of either Quartz PCI or analySIS but that I have had demos on both. I use a Shareware program called ThumbsPlus for my images, but I am guilty of the points that were made about not being as conscientious about putting the information into the database.
In my opinion, there was no violation of the rules of the Listserver and I agreed with the points that he was making. We just need to relax a little. I am making this plea because I was once "bitten" by a vendor who complained to Nestor that I was too enthusiastic in endorsing one company's product perhaps at the expense of theirs, where I had no commercial interest and that fact was known to the vendor. Where appropriate, Nestor steps in a makes personal comments to individuals when he thinks that they overstep the bounds. I would ask everyone to make certain that a lister's response was intended to "Tick" them off or "Make them tired" before going overboard. Just re-read a response first and think it over before criticizing too harshly. This is a valuable resource with people that are truly interested in helping their colleagues when they can and I hate reading the types of responses that prompted me to respond to this one.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (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: Yves Giroux [mailto:ygiroux-at-istar.ca] Sent: Tuesday, February 26, 2002 7:22 PM To: Microscopy-at-sparc5.microscopy.com
Hi NESTOR'
I'm getting very tired of those "promotional-type" answers (this listserver had, if I remember well, some rules against this).
We also at HITACHI have a very powerful Digital Image Management System (including very sophisticated Database) called PCI (by Quartz Imaging) but we are NOT using this listserver to promote it
} } } Anton-Jan, } } We do not use such a program here. My only experience (albeit very limited) } is with two programs: Aldos "Fetch" and "Quartz PCI". Gatan promotes } (promoted?) Fetch. We have at least one copy. Quartz PCI, the 3rd party } imaging and image manipulation software marketed by Hitachi, contains an } archiving software. } } My opinion is that any archiving software requires more up-front work than } it is worth. We choose, for the most part, to keep good records on the } sample types pertaining to each work request and locate images based on } this information. Thus, we tend to spend our time ensuring that we can } locate our archived files. The LabLan has become very useful toward this } purpose. } } Another source to check is the microscopy list server } www.msa.microscopy.com/MicroscopyListserver. One can search their } discussion threads for information on a variety of topics. One should note } that, for the most part, the comments on the list server are opinion. } Remember that opinions are like noses: everyone has one and my having one } is perfectly obvious to everyone else. } } Good luck and give my best to Johan and the rest of the MCM gang. } } Gary M. Brown } ExxonMobil Chemical Company } Baytown Polymers Center } 5200 Bayway Drive } Baytown, Texas 77520-2101 } phone: (281) 834-2387 } fax: (281) 834-2395 } e-mail: Gary.M.Brown-at-ExxonMobil.com } } } } } Alistair D } } Westwood To: Gary M } Brown/Baytown/ExxonMobil-at-xom, Jerry } W Ball/Baytown/ExxonMobil-at-xom, } Sandra M } Wapp/Baytown/ExxonMobil-at-xom, } David W } 02/26/02 06:55 Abmayr/Baytown/ExxonMobil-at-xom } } AM cc: } } Subject: image management } software } } } } } } Folks, } } Any comments on Anton-Jan's email. } } Ali } } Alistair D. Westwood } Team Leader - Microscopy & Surface Science } Materials Characterization Lab - Polymer Science Division } ExxonMobil Chemical Company } Baytown Polymers Center } 5200 Bayway Drive } Baytown, TX 77520 } } Ph: (281) 834-5741 } Fax: (281) 834-1793 } ----- Forwarded by Alistair D Westwood/Baytown/ExxonMobil on 02/26/02 06:54 } AM ----- } } } Anton-Jan Bons } } To: Alistair D } } Westwood/Baytown/ExxonMobil-at-xom, } Mark M } 02/26/02 01:14 Disko/East-US/ExxonMobil-at-xom } } AM cc: Johan } Stuyver/Benelux/ExxonMobil-at-xom, Marc } H Anthonis/Benelux/ExxonMobil-at-xom } } Subject: image management } software } } } } } } Mark, Ali, } We are looking for a software package to manage our microscopy images. We } have PhotoShop, but that's not ideal for batch file conversions, browsing } through large numbers of images, etc. We have a demo version of LView Pro ( } http://www.lview.com/AboutLViewPro.htm) which looks very convenient; it's
} only 50$ but of course we canot order it just like that... Do you use } software for image management? Do you have any suggestions? It would be } best if we all use the same software. } } Thanks. Regards, } Anton-Jan Bons } ExxonMobil Chemical - European Technology Center } Hermeslaan 2, B-1831 Machelen, Belgium } tel: +32 2 722 2838, fax: +32 2 722 2461 }
Hello, I've received a few quotations for EDX detectors and will put together a package of recommendations to our users. The one's I've narrowed it to are from EDAX, PGT, or IXRF. They both offer EDX systems for our cold FESEM Hitachi S5000. I was wondering if anyone has good/bad experiences they could relate, or suggestions for EDX detectors for this particular SEM.
Thanks for your help. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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
} Has anyone out there had any success with high pressure } freezing/freeze substitution of plant leaves for TEM examination? If } so, please contact me as I haven't. } } Bob
Bob,
We haven't done any HPF on leaf here, but have had great success on cambium and developing xylem. I think the greatest obstacle would be to fill the intercellular spaces with cryoprotectant. (See Fred Monson's concerns.) We use 0.2M sucrose in water for most plant tissues. If you immerse the cut tissues in cryoprotectant then place them under vacuum several times, you may have some success.
As to cryosubstitution, the best and easiest medium is (seems to be) 2% osmium tetroxide in acetone with 8% dimethoxypropane (as water scavenger). Substitute for at least 5 days for best results. (There are many other recipes depending on what your ultimate goal is.)
You may also ruin your hard work by adding resin too quickly during embedding.
If you have any questions, email me directly.
Kim ------- Kim Rensing Ph.D. Dept. of Botany, UBC 6270 University Blvd Vancouver, BC, Canada V6T 1Z4
I don't really see that much pushing of one product over another. What is being discussed are detailed technical issues. A buyer can use this information to sort our what is best for them.
The more data a buyer has, the better. Don't you agree? Or, are you saying that your product is the only viable option? Of course not. This is I think, a discussion forum. there are many facets to discussion.
gary g.
At 04:22 PM 2/26/2002, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In my opinon the latest thread on Image Management Software was within both the intent and letter of our rules.
Let me remind you all of the nominal rules about the use of the Listserver with respect to Commerical Products. This information is taken directly from the FAQ page which you ALL received a copy of when you subscribed, and is available on-line on the WWW site., the link to which is on EVERY EMAIL sent through this Listserver
As Scott W. pointed out. When I notice someone going over the line they get a warning. If it happens multiple times, they run the risk of becoming banned from posting messages to the Listserver.
Nestor Your Friendly Neighborhood SysOp
**************************************************** Exerpts from the Microscopy Listserver FAQ.... **************************************************** ------------------------------------ I am a Commerical Manufacturer. Can I participate? -------------------------------------
YES!
If you are a manufacturer, you are always welcome to observe/join in any discussion at all times. We do ask that everyone, please refrain from overt sales pitches and/or commericalism. If a product which you produce/sell can solve a problem or answer a question raised by anyone on this list, then by all means feel free to say so. Try to be brief about the product, state the simple facts in a few (short) sentences and then offer to continue the discussion with any interested parties offline or point people to a WWW Site with detailed information, so that individuals can download/access the relevant information. Usually it will be sufficient to just add your phone number and/or Email address to the end of your message, and you'll be contacted by anyone that is interested.
Please note that: UNSOLICITED product announcements are advertising and do not belong in this forum.
Remember, please keep your comments about any product you "sell" to a minimum.
It is not out of line to provide your company name, Email address or WWW site as part of your signoff/signature line, at the end of ANY message you post to this system.
This Listserver operates on the honor system with respect to to posting of advertising, so please respect these simple ground rules.
------------------------------------------------------------ Can I post a For Sale / Advertisement / Commerical / Marketing Survey Message(s)? ------------------------------------------------------------
No, that does not fit within the bounds of this discussion forum.
This listserver is not intended to be a Sales/Marketing/Survey mechanism for organizations, but rather it is an open discussion area about microscopy and microanalysis problems and solutions.
If you are an organization and have equipment you wish to donate, (or sell, for nominal cost i.e. no profit) then this is generally an acceptable posting. If you are not sure then send a copy of the announcement in question to Zaluzec-at-MSA.Microscopy.Com and I will give you my opinion. An example of this type would be an old decommissioned instrument which someone is trying to give away for removal/shipping costs, that would fit within the bounds of the purposes of this list.
There is a mechanism to post Commerical NEWS to the Microscopy Community. If you have something of a purely Commerical Nature and wish to make it known to the community you may FREELY use the following WWW site (just follow the on-line instructions)
I typically move about 15 SEMs per year. I like to have one source for the SEM move & rigging as I don't want any "finger pointing" if something goes wrong.
The Company that I have had the most sucess with Domestic or International Is:
Fast Forward Pamela Parsons (877) 978-6300
They will handle the rigging, crating & move. For dis-assembly of the SEM you may wish to call JEOL or a third party vendor.
Disclaimer: I have no vested interest in Fast Forward other than seeing a good Company be rewarded by recomendations.
Regards,
Earl Weltmer
Original Message: ----------------- } From: Mary Mager mager-at-interchange.ubc.ca
Dear David, You might have some luck contacting a piano moving company, if you don't have a company that speciallizes in moving hi-tech equipment. The piano movers are used to getting heavy, delicate instruments out of awkward places. At 04:25 PM 2/25/2002 -0500, you 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 -----------------------------------------------------------------------. } } Greetings, } } I needed a company to move a Joel SEM with EDX from central New Jersey to Cleveland Ohio. } } The Cleveland Museum of Natural History has had an offer to have a Joel SEM donated. The catch is that it is in the basement of a consultants house. Does anyone know of a company in the Ohio-Pennsylvania-New Jersey (New York?) area who is, at the least, capable of hoisting it out of his basement and onto a truck? We may also be interested in delivery and re-assembly and alignment. } } Thank you for any suggestions, } Dr. David Saja, Geologist } dsaja-at-cmnh.org 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
-------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ .
I typically move about 15 SEMs per year. I like to have one source for the SEM move & rigging as I don't want any "finger pointing" if something goes wrong.
The Company that I have had the most sucess with Domestic or International Is:
Fast Forward Pamela Parsons (877) 978-6300
They will handle the rigging, crating & move. For dis-assembly of the SEM you may wish to call JEOL or a third party vendor.
Disclaimer: I have no vested interest in Fast Forward other than seeing a good Company be rewarded by recomendations.
Regards,
Earl Weltmer
Original Message: ----------------- } From: Mary Mager mager-at-interchange.ubc.ca
Dear David, You might have some luck contacting a piano moving company, if you don't have a company that speciallizes in moving hi-tech equipment. The piano movers are used to getting heavy, delicate instruments out of awkward places. At 04:25 PM 2/25/2002 -0500, you 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 -----------------------------------------------------------------------. } } Greetings, } } I needed a company to move a Joel SEM with EDX from central New Jersey to Cleveland Ohio. } } The Cleveland Museum of Natural History has had an offer to have a Joel SEM donated. The catch is that it is in the basement of a consultants house. Does anyone know of a company in the Ohio-Pennsylvania-New Jersey (New York?) area who is, at the least, capable of hoisting it out of his basement and onto a truck? We may also be interested in delivery and re-assembly and alignment. } } Thank you for any suggestions, } Dr. David Saja, Geologist } dsaja-at-cmnh.org 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
-------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ .
I typically move about 15 SEMs per year. I like to have one source for the SEM move & rigging as I don't want any "finger pointing" if something goes wrong.
The Company that I have had the most sucess with Domestic or International Is:
Fast Forward Pamela Parsons (877) 978-6300
They will handle the rigging, crating & move. For dis-assembly of the SEM you may wish to call JEOL or a third party vendor.
Disclaimer: I have no vested interest in Fast Forward other than seeing a good Company be rewarded by recomendations.
Regards,
Earl Weltmer
Original Message: ----------------- } From: Mary Mager mager-at-interchange.ubc.ca
Dear David, You might have some luck contacting a piano moving company, if you don't have a company that speciallizes in moving hi-tech equipment. The piano movers are used to getting heavy, delicate instruments out of awkward places. At 04:25 PM 2/25/2002 -0500, you 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 -----------------------------------------------------------------------. } } Greetings, } } I needed a company to move a Joel SEM with EDX from central New Jersey to Cleveland Ohio. } } The Cleveland Museum of Natural History has had an offer to have a Joel SEM donated. The catch is that it is in the basement of a consultants house. Does anyone know of a company in the Ohio-Pennsylvania-New Jersey (New York?) area who is, at the least, capable of hoisting it out of his basement and onto a truck? We may also be interested in delivery and re-assembly and alignment. } } Thank you for any suggestions, } Dr. David Saja, Geologist } dsaja-at-cmnh.org 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
-------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ .
I typically move about 15 SEMs per year. I like to have one source for the SEM move & rigging as I don't want any "finger pointing" if something goes wrong.
The Company that I have had the most sucess with Domestic or International Is:
Fast Forward Pamela Parsons (877) 978-6300
They will handle the rigging, crating & move. For dis-assembly of the SEM you may wish to call JEOL or a third party vendor.
Disclaimer: I have no vested interest in Fast Forward other than seeing a good Company be rewarded by recomendations.
Regards,
Earl Weltmer
Original Message: ----------------- } From: Mary Mager mager-at-interchange.ubc.ca
Dear David, You might have some luck contacting a piano moving company, if you don't have a company that speciallizes in moving hi-tech equipment. The piano movers are used to getting heavy, delicate instruments out of awkward places. At 04:25 PM 2/25/2002 -0500, you 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 -----------------------------------------------------------------------. } } Greetings, } } I needed a company to move a Joel SEM with EDX from central New Jersey to Cleveland Ohio. } } The Cleveland Museum of Natural History has had an offer to have a Joel SEM donated. The catch is that it is in the basement of a consultants house. Does anyone know of a company in the Ohio-Pennsylvania-New Jersey (New York?) area who is, at the least, capable of hoisting it out of his basement and onto a truck? We may also be interested in delivery and re-assembly and alignment. } } Thank you for any suggestions, } Dr. David Saja, Geologist } dsaja-at-cmnh.org 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
-------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ .
I didn't answer this earlier, as have only tried HPF a couple of times. The key is, as Fred Monson implies, to fill all air spaces with liquid first. I've done this by vacuum infiltration of pieces of mature root, but have not tried leaf pieces. Some people also surround the tissue with some sort of cryoprotectant, or at least with some (usually fairly viscous) solution that is less likely to form crystals, like a solution of dextrin or BSA, for example. You could try vacuum infiltration of the leaf pieces, either with water or with some aqueous non-osmotic "cryoprotectant". Good luck, cheers, Rosemary
} } Morning Bob, } I hope this is on point, but as a zoologist, I would like to know } how the mesophyl (?-no elementary bio book to check on leaves anymore) holds } up at 30K psi? You may have the same problem with a leaf as I might have } with a piece of mammalian lung. At 30K psi I can only imagine that the } piece of lung would simultaneously be frozen and compressed - to me, that } means, destroyed. } Most of this has been imagined, so I won't spend any more time on } it. Interestingly, the only botany book I have at the moment in my library } is by Johansen, D.A, Plant Embryology, 1950 (Cycads to Anthophyta???- } Spermatophyta (of the time?)), and, of course, no leaves in it. } } Regards, } } Fred Monson } } } ---------- } } From: Bob Wise } } Sent: Monday, February 25, 2002 12:39 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: HPF of leaf tissue } } } } ------------------------------------------------------------------------ } } 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 out there had any success with high pressure } } freezing/freeze substitution of plant leaves for TEM examination? If } } so, please contact me as I haven't. } } } } Bob } } } } -- } } Robert R. Wise, Ph.D. } } Associate Professor of Plant Physiology } } Department of Biology and Microbiology } } University of Wisconsin Oshkosh } } } } On sabbatical from Sept. 2001 to Aug. 2002 at UW Madison } } Botany Department } } B217 Birge Hall } } 430 Lincoln Drive } } Madison, WI 53706 } } (608) 262-4288 (phone) } } (608) 262-7509 (fax) } } wise-at-uwosh.edu } } http://www.wisc.edu/biotron/Sharkey/ } } http://www.uwosh.edu/departments/biology/wise/wise.html } } } }
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
61- 2 6246 5475 or 61- 0402 835 973 rosemary.white-at-csiro.au
I typically move about 15 SEMs per year. I like to have one source for the SEM move & rigging as I don't want any "finger pointing" if something goes wrong.
The Company that I have had the most sucess with Domestic or International Is:
Fast Forward Pamela Parsons (877) 978-6300
They will handle the rigging, crating & move. For dis-assembly of the SEM you may wish to call JEOL or a third party vendor.
Disclaimer: I have no vested interest in Fast Forward other than seeing a good Company be rewarded by recomendations.
Regards,
Earl Weltmer
Original Message: ----------------- } From: Mary Mager mager-at-interchange.ubc.ca
Dear David, You might have some luck contacting a piano moving company, if you don't have a company that speciallizes in moving hi-tech equipment. The piano movers are used to getting heavy, delicate instruments out of awkward places. At 04:25 PM 2/25/2002 -0500, you 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 -----------------------------------------------------------------------. } } Greetings, } } I needed a company to move a Joel SEM with EDX from central New Jersey to Cleveland Ohio. } } The Cleveland Museum of Natural History has had an offer to have a Joel SEM donated. The catch is that it is in the basement of a consultants house. Does anyone know of a company in the Ohio-Pennsylvania-New Jersey (New York?) area who is, at the least, capable of hoisting it out of his basement and onto a truck? We may also be interested in delivery and re-assembly and alignment. } } Thank you for any suggestions, } Dr. David Saja, Geologist } dsaja-at-cmnh.org 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
One researcher here is doing research on the properties of thin films. Part of the studies would be to do TEM imaging of the films. What type of support film can we put on Cu grids for the evaporated film. I had a go with formvar, but they brake down during evaporation due to the heat of the plasma. I would like to dissolver the support film if possible afterwards.
Suggestions?
Mr. S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gabarone Botswana
Mr. S. H. Coetzee wrote the following: ================================================================ One researcher here is doing research on the properties of thin films. Part of the studies would be to do TEM imaging of the films. What type of support film can we put on Cu grids for the evaporated film. I had a go with formvar, but they brake down during evaporation due to the heat of the plasma. I would like to dissolver the support film if possible afterwards. ================================================================ There are several possibilities here but it is not clear what you mean by "during evaporation due to the heat of the plasma".
If Formvar® is not acceptable, for whatever the reasons, you could consider carbon or if you really are exposing these to a plasma, then SiO2 films. Information about these films can be found on URL http://www.2spi.com/catalog/grids/cusctgrd.html However, both films exhibit a certain degree of granularity on the nanoscale which might be distracting if not also confusing in the analysis of your micrographs.
Another approach is to pick up the films on a silicon nitride membrane window grid, see URL http://www.2spi.com/catalog/instruments/silicon-nitride.html The window is not going to be hurt by exposure to an oxygen plasma, and it is completely amorphous and therefore structureless and featureless. It is also quite electron transparent.
There is a final possibility for solving the problem. You did not mention what mesh size you are using, but you switch to using 2000 mesh grids and possibly eliminate the need altogether for the use of any support film or the more expensive silicon nitride membrane window grids.
Disclaimer: SPI Supplies offers custom coated grids and the other items mentioned in this posting.
Chuck
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We regularly sputter or evaporate thin films onto standard carbon filmed (no formvar) Cu or Au grids, usually 200 mesh. We have built a small holder to mount the grids into for ease of handling. Don't put the evaporation source too close to the grids, I think ours is about 150mm.
Good luck, Ron
On Wed, 27 Feb 2002 11:16:32 +0200 "Coetzee, Mr S. H Physics Science" {COETZEES-at-mopipi.ub.bw} 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 } -----------------------------------------------------------------------. } } } Dear all } } One researcher here is doing research on the properties of thin films. Part } of the studies would be to do TEM imaging of the films. What type of } support film can we put on Cu grids for the evaporated film. I had a go } with formvar, but they brake down during evaporation due to the heat of the } plasma. I would like to dissolver the support film if possible afterwards. } } Suggestions? } } } Mr. S. H. Coetzee } Electron Microscope Unit } University of Botswana } Private Bag 0022 } Gabarone } Botswana } } Phone : +267 355 2426 } Mobile: +267 718 96 729 } Fax : +267 585 097 } e-mail : coetzees-at-mopipi.ub.bw {mailto:coetzees-at-mopipi.ub.bw} } } }
---------------------- 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
well peter considering you are the compnay i was refering to, then it must have just been one of those weeks for you. peter i have delt with you directly and the truth is i find the manufactures to have much better trained people. i wasn't going to mention it, but you decided to put your two cents in and get free advertisement on the list server.
in a dept i worked at about 9 years ago, we bought a used 301 from you. you came in installed it got a vacuum and said it was running fine.i come in the next day the scope is un alinged. so i did that within 3 days the scope was down. it took us over a week to get you back to get it up and running. we then called philips and had it put unnder their contract. if you want to respond email me directly. John Hoffpauir Thomas Jefferson University
All information given here is based on experience.
I have been using Pesto, Inc. for the last four years. JEOL was getting too expensive, especially since I have two scopes. The response time from Pesto, Inc. has been great and getting replacement parts has not been a problem.
Lou Bustillos AMA Analytical Services, Inc.
---------- Original Message ---------------------------------- } From: "John Hoffpauir" {John.Hoffpauir-at-mail.tju.edu}
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has anyone out there had any experience in cutting one micron sections of transwell membranes? a tech in my lab is having trouble getting them flat on a glass slide. he has wrinkles which is unacceptable for publication. i have never worked with them. any advice would be great. john hoffpauir
{paraindent} {param} right {/param} {color} {param} 0100,0100,0100 {/param} {/paraindent}
{paraindent} {param} right {/param} Hello all, {/paraindent}
{paraindent} {param} right {/param} {/color} We are attempting to perform time-lapse imaging of GFP-labelled cells on a Zeiss Axiophot 1 (an upright microscope). There are two aspects of the system that cause us trouble. The first is our Uniblitz shutter, which is driven via parallel port by a module that is part of the Universal Imaging Metamorph v.4.0 suite. The shutter responds to oscilloscope-generated current pulses to provide open (exposure) times of 20 ms (as stated in the specs). We require open times of 100-200 ms, but the shortest pulse interval that can be generated through the parallel cable by any exposure control function in the software is 500- 600 ms. No-one connected with U.I. or the company that sold us our system can explain why this is the case, and how we might achieve shorter exposure times. Is there anyone out there with experience or knowledge that might be brought to bear on this issue? {/paraindent}
{paraindent} {param} right {/param} The second problem is more troubling. Even without a heating chamber, we experience a small, variable, but problematic downward drift of the specimen stage. The total drift is ~0.5 -1um per minute (sometimes more or less), but is enough to scuttle velocity measurements with our 1.4 NA lens. To solve this, we bought a Prior Optiscan z-axis focus controller, which meshes a gear with the microscope's coarse focus gear. We thought this might prevent the subtle stage slippage that is compromising our observations. No dice. Again, does anyone have an insight, knowledge or words of wisdom? Thanks in advance. {/paraindent}
If your polymer films are breaking, you might be better off if you first evaporate C on them for stability, then on the opposite side of the grid deposit your Al. Finally one can remove the carbon/polymer in a proper solvent leaving the Al film all by itself. Alternatively, one can deposit the Al films on rocksalt and float the films off in water.
I hope this helps
Jordi
PS. We have often deposited carbon (not metal) on polymer films without problems, maybe you can place your grids/w.polymer further away form the Al source to minimize breakage. ---------------------- } Mr. S. H. Coetzee wrote: } } } Dear all } } One researcher here is doing research on the properties of thin films. Part } of the studies would be to do TEM imaging of the films. What type of } support film can we put on Cu grids for the evaporated film. I had a go } with formvar, but they brake down during evaporation due to the heat of the } plasma. I would like to dissolver the support film if possible afterwards. } } Suggestions? } } } Mr. S. H. Coetzee } Electron Microscope Unit } University of Botswana } Private Bag 0022 } Gabarone } Botswana } } Phone : +267 355 2426 } Mobile: +267 718 96 729 } Fax : +267 585 097 } e-mail : coetzees-at-mopipi.ub.bw {mailto:coetzees-at-mopipi.ub.bw} } } }
---------------------- 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
Dear Mr. Coetzee, We use a carbon evaporated film to support thin films and small particles for our TEM work. First we cast a collodion film on water, then float TEM grids on it, lift the film with filter paper and dry it. Carbon coat the collodion on grids and then dissolve the colodion off in a Jaffe washer filled with chloroform for 48 hours. The resulting amorphous carbon film is strong and conductive and easily lasts 200 kV in our TEM. At 11:16 AM 2/27/2002 +0200, you wrote: } Dear all } } One researcher here is doing research on the properties of thin films. Part } of the studies would be to do TEM imaging of the films. What type of } support film can we put on Cu grids for the evaporated film. I had a go } with formvar, but they brake down during evaporation due to the heat of the } plasma. I would like to dissolver the support film if possible afterwards. } } Suggestions? } } } Mr. S. H. Coetzee 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
Dear Vitatly, I'm sorry I didn't post a link to the Quartz XOne EDS system. It is: www.quartzimaging.com. This product is not the same as the Quartz PCI image capture system and is not assiciated with Hitachi. Disclaimer: I have been involved with the design and selling of the XOne system since its inception. At 03:47 PM 2/26/2002 -0500, you wrote: } (snip) } } What he says about } } EDS is partially true but most EDS systems (including older ones) come } } with } } beam control packages. The advantage of getting the beam control from the } } EDS system is that they provide integrated software to do digital mapping, } } linescans, in addition to electron imaging. } } EDS system with the beam control package is an active system. With all } } To Mary Mager- thank you for the correction. I was not aware of EDS mapping } capabilities of the passive Quartz PCI system- can you post a link? } } Cheers. } } Vitaly Feingold
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
S.H. You could use a carbon support film. You can get quite thin using indirect deposition by evaporation thereby eliminating any contributing structure. A lacy film may be susceptible to damage as a support for the carbon so a high mesh grid would be appropriate. We often float films off cleaved mica used as a substrate although this depends on adhesion and may not be appropriate for your films. Good Luck, Russ Gillmeister Microscopy Bldg. 114-42D Xerox Corp. 800 Phillips Rd. Webster, NY 14580 (585) 422-5317
-----Original Message----- } From: Coetzee, Mr S. H Physics Science [mailto:COETZEES-at-mopipi.ub.bw] Sent: Wednesday, February 27, 2002 4:17 AM To: Listserver (E-mail)
Dear all
One researcher here is doing research on the properties of thin films. Part of the studies would be to do TEM imaging of the films. What type of support film can we put on Cu grids for the evaporated film. I had a go with formvar, but they brake down during evaporation due to the heat of the plasma. I would like to dissolver the support film if possible afterwards.
Suggestions?
Mr. S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gabarone Botswana
I recently carbon coated some samples for a user who took them to a microprobe for analysis.
He called to ask about the carbon coating's affect on the appearance of the sample after analysis.
On a previous run, with a different instrument, he said the beam 'toasted' the area of analysis and he liked that since it showed exactly where the beam had been. On the instrument he used here, there was no toasting, so no visible record of the analytical location.
I don't run the probe, I just do the carbon coating. But I thought someone out there in microscope land might know what I can tell him. He says the toasting probe was newer and better than the older one we have here. He said the beam conditions were similar for the two instruments.
I am curious. What are the toast tracks and does the carbon coating have anything to do with them?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
All plastics will break down from the heat after a period of time. A carbon substrate will last longer. When we make our carbon substrate we use carbon and formvar and then dissolve the formvar. If this doesn't solve your problems contact us we have other options.
John Arnott --
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Yash Agarwal wrote: } } Hi, } } I am an engineer at Ikonisys Inc. We were looking for a graticule with the } following requirements } } 1. Graticule for Reflected Fluorescence Imaging. (No bright field) } 1. Graticule slide dimensions - 75mm x 25mm x 1mm. } 2. Cross pattern Graticule with Micrometer scale of 5mm in 0.05mm divisions. } } Could you provide me with the information whether you would have such a } Graticule? } If not can it be custom made and if yes what would it cost? } } Thank you } } Yash Agarwal } Ikonisys Inc. } 5 Science Park } Suite 1000, } New Haven, CT 06515 } Tel: 203-776-0791 ext. 289 } Fax: 203-776-0795 } Email: yash.agarwal-at-ikonisys.com
Two questions, 1) How did he put the carbon on? If with a dirty system, he could be getting contamination. 2) What is his sample made of. An electron microprobe uses a large current and can damage some samples. Glass is a prime example.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: jmkrupp-at-cats.ucsc.edu [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Wednesday, February 27, 2002 12:59 PM To: Microscopy-at-sparc5.microscopy.com
Hi:
Sorry if this is a little off our topic line.
I recently carbon coated some samples for a user who took them to a microprobe for analysis.
He called to ask about the carbon coating's affect on the appearance of the sample after analysis.
On a previous run, with a different instrument, he said the beam 'toasted' the area of analysis and he liked that since it showed exactly where the beam had been. On the instrument he used here, there was no toasting, so no visible record of the analytical location.
I don't run the probe, I just do the carbon coating. But I thought someone out there in microscope land might know what I can tell him. He says the toasting probe was newer and better than the older one we have here. He said the beam conditions were similar for the two instruments.
I am curious. What are the toast tracks and does the carbon coating have anything to do with them?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
When we carbon coat samples, we too get those "toasted" areas that's believed to be due to carbon build up where the beam interacts with the specimen. We generally use 20KV with a beam current of 20nA. They can be helpful to see exactly where the analysis was done and the beam diameter as well. However, when the sample is mounted in conductive mounting material and no carbon coating is required, the "toasted" areas aren't nearly as visible if at all at the same operating parameters.
I need some information. I hope I will get some responses from you. I was wondering what the difference is between a SIMS Secondary Ion Mass Spectrometer and an Ion Beam Electron Microscope. Please excuse my ignorance, but I've tried looking on the web, and I haven't found the explanation that I need.
The toasting process is probably similar to the mechanism that causes contamination scan marks. The electron beam interacts with the carbon molecules and ionizes them. The react with each other to form amorphous polymers that are a visible to the secondary electron detectors. For uncoated specimens the "toasting" marks are just plain hydrocarbon contamination deposits. The beam is interacting with the residual hydrocarbon partial pressure to create ions that follow the e beam down to the surface to form that polymer goo.
Ronald Vane XEI Scientific anticontamination systems www.SEMCLEAN.com 650-369-0133
It's also difficult with EM sections. I've always assumed the wrinkling is due to the resin expanding and the membrane not. I haven't tried this, but dissolving the resin with ethoxide would probably fix the problem.
Diana
At 10:15 AM -0500 27/2/02, "JHoffpa464-at-aol.com"-at-sparc5.microscopy.com wrote: } } has anyone out there had any experience in cutting one micron } sections of transwell membranes? a tech in my lab is having trouble } getting them flat on a glass slide. he has wrinkles which is } unacceptable for publication. i have never worked with them. any } advice would be great. } john hoffpauir
--
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
The "toasted" appearance on the surface of the sample at the area/point of analysis is extra carbon that is deposited from the probe's inside vacuum environment. The source of this carbon is from the vacuumpumps, especially the rotary pump.
The longer the area is exposed to the electron beam, the higher the beam current, the more carbon is deposited on that spot or raster area.
These extra carbon deposited on your beam on the spot of analysis causes the matrix correction prosedures and standarizing procedures to be incorrect as the standards and unknown samples are normally carbon coated to a very precise thickness of 25 nanometers.
Best wishes, Leon
Skool vir Omgewingswetenskappe en Ontwikkeling: Geologie, GIS \\\\\\\\\\\\\\\\} URL: http://www.puk.ac.za } WWW} \\\\\\\\\\\\\\\\\\\ Leon Smuts-Electronmicroprobe-Potchefstroom University-South Africa /////////////} mailto: plbls-at-puknet.puk.ac.za } eMail} ///////////////
} } } Jon Krupp {jmkrupp-at-cats.ucsc.edu} 02/27/02 07:59PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi:
Sorry if this is a little off our topic line.
I recently carbon coated some samples for a user who took them to a microprobe for analysis.
He called to ask about the carbon coating's affect on the appearance of the sample after analysis.
On a previous run, with a different instrument, he said the beam 'toasted' the area of analysis and he liked that since it showed exactly where the beam had been. On the instrument he used here, there was no toasting, so no visible record of the analytical location.
I don't run the probe, I just do the carbon coating. But I thought someone out there in microscope land might know what I can tell him. He says the toasting probe was newer and better than the older one we have here. He said the beam conditions were similar for the two instruments.
I am curious. What are the toast tracks and does the carbon coating have anything to do with them?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I have been in charge recently of our SEM lab, in which there is a rather idle JEOL5400LV (with a Kevex EDS) microscope. According to the person who is in charge of running this machine, the capabilities of this SEM are restricted to study uncoated dry samples, preferably mineral. Does anybody have further experience on this machine (or an upgraded one?) like imaging wet samples, etc.?
I have some suggestions for the second problem you mention. I have used our own in-house developed multi-position time-lapse system for years and I have experienced the same problems. We used a system with an incubation chambre in which we could keep cells on 37 degrees centigrade or on room temperature (18 - 21 degrees centigrade). We also noticed a small drift which was specially annoying on high N.A. lenses as you too have noticed.
After some experiments we foud out that is was caused by the response of the metal of the microscope to changes in the equipment temperature. We often did 10 time-lapse movies in parallel on one micrscope over the weekend and we could monitor that the changes in the room temperature had an influence on the position of the motorised stage (X,Y and Z -axis). After we found out what caused the problem we simply refocused for each position at each time-lapse cycle, which solved the problem for us.
Another option could be to put the entire microscope in a temperature controlled environment. In most temperature control system, only part of the microscope is inside the incubation chamber, all the rest is exposed to the changes of the room temperature. Even in a modern building, the air conditioning system is not precise enough to stabilise the temerature enough for high-resolution microscopy.
Best regards,
Peter -- Dr. Peter Van Osta
Union Biometrica N.V./S.A. European Scientific Operations
Cipalstraat 3 B-2440 Geel Belgium
tel.: +32 (0)14 570 619 fax.: +32 (0)14 570 621
http://www.unionbio.com/
================================ } Hello all, } We are attempting to perform time-lapse imaging of GFP-labelled cells on a Zeiss Axiophot 1 (an upright microscope). There are two aspects of the system that cause us trouble. The first is our Uniblitz shutter, which is driven via parallel port by a module that is part of the Universal Imaging Metamorph v.4.0 suite. The shutter responds to oscilloscope-generated current pulses to provide open (exposure) times of 20 ms (as stated in the specs). We require open times of 100-200 ms, but the shortest pulse interval that can be generated through the parallel cable by any exposure control function in the software is 500- 600 ms. No-one connected with U.I. or the company that sold us our system can explain why this is the case, and how we might achieve shorter exposure times. Is there anyone out there with experience or knowledge that might be brought to bear on this issue? } The second problem is more troubling. Even without a heating chamber, we experience a small, variable, but problematic downward drift of the specimen stage. The total drift is ~0.5 -1um per minute (sometimes more or less), but is enough to scuttle velocity measurements with our 1.4 NA lens. To solve this, we bought a Prior Optiscan z-axis focus controller, which meshes a gear with the microscope's coarse focus gear. We thought this might prevent the subtle stage slippage that is compromising our observations. No dice. Again, does anyone have an insight, knowledge or words of wisdom? Thanks in advance. } } .
We haven't cut these types of membranes, but when we thick section large windowed plastics they tend to wrinkle on slides. What we do is float them onto a drop of water on the slide and heat on a hot plate to evaporate the water. Once thoroughly dry, sections are stained,rinsed and immediately coversliped with permount (careful not to move in the xy axis when lightly compressing). This usually prevents section wrinkles.
Good luck
Mike D. JHMI Microscope Facility
P.S. slower sectioning and smaller faces may help.
} ------------------------------------------------------------------------ } 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 out there had any experience in cutting one micron sections of transwell membranes? a tech in my lab is having trouble getting them flat on a glass slide. he has wrinkles which is unacceptable for publication. i have never worked with them. any advice would be great. } john hoffpauir
} } has anyone out there had any experience in cutting one micron } sections of transwell membranes? a tech in my lab is having trouble } getting them flat on a glass slide. he has wrinkles which is } unacceptable for publication. i have never worked with them. any } advice would be great. } john hoffpauir *************************** I work wit them a lot....they always wrinkle. I have tried vapors, heat, drying them down slowly on a large water drop,drying them down quickly on a small water drop...you get the picture. I can offer no solution for your tech, just sympathy. My clients usually opt for taking a high mag shot of a relatively flat area. Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
An advanced course on polarized light microscopy which will cover the following topics:
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{color} {param} 0100,0100,0100 {/param} Hello all, {FontFamily} {param} Arial {/param}
{/color} {FontFamily} {param} Times New Roman {/param} I am once again in awe of the marriage of expertise and generosity on this listserver. Thank you all for your replies. Since yesterday, I have received many replies and would particularly like to thank Dan Focht of Bioptechs for a lengthy telephone session. Here's a progress report so far. I neglected to mention in my original post (sorry!) that, in order to rule out thermal effects from our heating chamber, we began to run time-lapse series on microscope slides of fixed cells (no chamber). Since the chamber is quite heavy, we also eliminated stage drift due to this added weight. The drift was still present. As many of you have pointed out, the location of heating/cooling vents, heat-generating equipment and even exothermic grad students might influence focus. We have reasoned that thermal instability should shift the focal plane both above and below the specimen plane and in an almost random fashion. The focal plane always shifts to a point above the specimen (consistent with the stage lowering). In addition, the rate at which this occurs is more or less reproducible, regardless of the number of bodies present or milling about in the room and of the average room temperature (though we haven't monitored the stage, objective or slide temperature systematically). So, we have tentatively concluded that the problem is related to the specific focus mechanism of our microscope. In conjunction with our local Zeiss service rep, we will be attempting a solution based on this conclusion. I won't describe it at this time since it may only be applicable to a small number of microscopes and probably shouldn't be undertaken by people (like me) without a detailed understanding of the focusing mechanism of the microscope in question. Our service rep has reluctantly agreed to try a simple alteration of the mechanism. I'll let you know if it works and what it was, if it does. As to the shutter open time... Many of you have suggested plausible causes for the problem. I have tried many of the "common-sense" solutions suggested (e.g., removing unnecessary peripherals, resident programs and tweaking settings in MetaMorph's Device and Drop-in Manager systems). Other aspects of system performance have improved. More than one of you has suggested construction of a one shot, leading edge-triggered TTL pulse generator. This seems to me to be the best way to get the shutter to do what it's told. I'll keep you posted on this as well. Thank you all again.
I have found the best way to cut Transwell and other membranes is to orient the membrane parallel to the knife edge (horizontal) instead of the conventional vertical orientation. For 1 micrometer and 70nm sections, use a slow cutting speed (0.5mm/s or less) and you should be able to produce chatter-free sections. The investigators that I work with usually want to see migration of the cells through the membrane pores plus the cell layers on both sides of the membrane, and I find cutting in this manner produces better results (not to say that I don't get a wrinkle or two now and then). For thick sections, the old fashioned gelatin-chrome alum coated slides work better than poly-lysine or Plus slides--the sections adhere well and usually remain wrinkle-free after staining with toluidine blue. I also embed the membranes in the harder formulation of Spurr's resin and highly recommend using a diamond knife for both thick and ultrathin sectioning. -- Victoria J. Madden Microscopy Services Laboratory Pathology and Laboratory Medicine University of North Carolina at Chapel Hill
An individual new to EM wanted to learn from me how to formvar-carbon coat slotted grids. The person bought grids made of a Berylium-Copper alloy. I called the vendor and was warned about the danger of Berylium and it's vapor. I told the invidual to get copper grids for our use. Now in all my years in EM I have not worked with Berylium. Would someone out there send me some information on the hazards of Berylium? Also what applications are Berylium-Copper or Berylium only grids used for? Any information is appreciated, thanks.
Tom Bargar EM Lab, UNMC tbargar-at-unmc.edu 402-559-7347
Hello Oliver, We have a JEOL 5900LV in our lab, which I think is a minor update/upgrade to the machine you have. The "LV" designation typically means that the microscope is capable of "low vacuum" operation. In our case, we didn't purchase that option, even though we have that model number designation on our scope. You will probably need to check your documentation to verify if you have that feature, and how to use it. You might also want to check with the local JEOL service/sales people who sold you the instrument to determine its capabilities (http://www.jeol.com). We use ours quite heavily for typical high vacuum SEM purposes, and it performs well for a standard tungsten filament SEM. Most of our work has been with glasses, ceramics, metals, and polymers. When you say "wet imaging", that typically refers to an evironmental SEM, which is a more sophisticated type of SEM (lenses, detectors, and vacuum pumping system modified to accomodate chamber conditions that are different than near the gun.). Good luck, and enjoy!
-Brad
---------- From: Legendre Olivier Sent: Thursday, February 28, 2002 12:02 AM To: 'Microscopy-at-MSA.Microscopy.Com' Subject: JEOL5400LV
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I have been in charge recently of our SEM lab, in which there is a rather idle JEOL5400LV (with a Kevex EDS) microscope. According to the person who is in charge of running this machine, the capabilities of this SEM are restricted to study uncoated dry samples, preferably mineral. Does anybody have further experience on this machine (or an upgraded one?) like imaging wet samples, etc.?
Dear Jonathan, The marks that a microprobe can make on a sample surface can come from many sources, but a clean, evaporated carbon coat should not be one of them. The microprobe itself can lay down contamination from its own vacuum system, the sample can evolve contamination that is carbonized in the high-current beam or other sources of contamination might be present. One of the sources of contamination I found when I did a study of contamination rates and contamination clean-off by air-jet, was that an incidence of the beam hitting the mounting epoxy at the edge of a sample caused increased contamination in the instrument for three days. If you scan or test at the edge of a mounted specimen you can see the marks of the scan in the light microscope, where the epoxy has been boiled away by the beam. I would say that the absence of the toast marks in your microprobe was a good thing and indicates your vacuum system is better than the other one he used, even if he likes these marks. At 09:59 AM 2/27/02 -0800, you wrote: } Hi: } } Sorry if this is a little off our topic line. } } I recently carbon coated some samples for a user who took them to a } microprobe for analysis. } } He called to ask about the carbon coating's affect on the appearance of } the sample after analysis. } } On a previous run, with a different instrument, he said the beam 'toasted' } the area of analysis and he liked that since it showed exactly where the } beam had been. On the instrument he used here, there was no toasting, so no } visible record of the analytical location. } } I don't run the probe, I just do the carbon coating. But I thought someone } out there in microscope land might know what I can tell him. He says the } toasting probe was newer and better than the older one we have here. He } said the beam conditions were similar for the two instruments. } } I am curious. What are the toast tracks and does the carbon coating have } anything to do with them? } } Thanks } } Jonathan Krupp
Regars, 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-interchange.ubc.ca
This is a beautiful low (lousy) vacuum SEM. Its sounds like the "restricted to study uncoated dry samples, preferably mineral" was put on the microscope by someone at the SEM Lab who didn't want any other samples/users using the scope.
I've often seen where the "Materials People" and the Geology people fobid messy unstable biological samples in their EM's in order to keep the scopes clean (particularly when dealing with analytical systems). (Now, where do the clays and rubber people fit in??)
Now, before you get someone upset you might find out the history of the "restricted to study uncoated dry samples, preferably mineral" and who paid for the scope (got the grant). But an EM is a terrible thing to waste.
On 28 Feb 2002, at 9:02, Legendre Olivier wrote:
} Hi all, } } I have been in charge recently of our SEM lab, in which there is a rather } idle JEOL5400LV (with a Kevex EDS) microscope. According to the person who } is in charge of running this machine, the capabilities of this SEM are } restricted to study uncoated dry samples, preferably mineral. } Does anybody have further experience on this machine (or an upgraded one?) } like imaging wet samples, etc.? } } Thank you for helping } } Olivier Legendre } BRGM } 3, avenue C. Guillemin } 45060 ORLEANS CEDEX 2 FRANCE } Tel: (33) 0 238 64 38 03 } Fax: (33) 0 238 64 37 11 } e-mail: o.legendre-at-brgm.fr } } } } } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
I would be interested to hear from anyone who is still using a Kevex Sesame24/Microspec 2A WDS system. If you have one that is retired, I would especially have an interest in the possibility of obtaining spare cards. Mine is working but have no spares...
Another question about the Sesame... I no longer have the Kevex 8000/Delta EDS to which the WDS sent data for quantitation. Has anyone mated the Sesame to a different EDS/software package to do quantitation?
Thanks, Woody -------------- Woody White McDermott Technology, inc nwwhite-at-mcdermott.com
We have a Hitachi LV here. We go back and forth between high vacuum mode and low vacuum quite a bit. We have a JEOL 840A down the hall and try to steer people there if they really don't need the low vacuum mode. It is simply a matter of balancing the use.
I am not familiar with the 5400 capabilities. If it is like ours, it is not a true environmental scope. We can only manage an atmosphere of 2 Torr (270 Pa) which is not enough to keep wet samples wet for very long. Also, the detectors and such are not optimized for true environmental mode. But it does great for insulating specimens.
At 09:02 AM 2/28/02 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This is not a "reply" to the post, but perhaps an extension of the question.
Why would Cu-Be grids be used? Cu-Be is an interesting and quite widely-used class of alloys (see http://microstructure.copper.org), but I don't see why it would be used for EM grids - unless, that is, the grids we loosely call "copper" are in fact all made of Cu-Be?
On the subject of the posting, I can't imagine that Cu-Be would be as widely used as it is if it were as toxic as Be-metal. I'm sure the alloying must drastically reduce the hazard, but this is not my area of expertise.
Tony
At 10:31 AM 2/28/2002 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have two Microm HM505 cryostats: one HM505N (non-motorized advance) and one HM505E (motorized advance). We've not had too much trouble with them, considering we've had one for at least five years and the other, which was recently donated to us, is probably older. The few problems we've experienced with the one we purchased have been outside the norm and were resolved relatively quickly (this doesn't take into consideration operator error -- an inexperienced user ran the specimen chuck into the knife holder and put a large dent in it).
Both units section well. In fact, the batch of blocks I cut on the HM505E earlier this week were probably the best I've ever cut (and we still have an old IEC workhorse around here somewhere).
I believe Microm is carried by Richard-Allan Scientific and distributed by Fisher.
Hope this helps you out,
Jaclynn M. Lett jlett-at-cid.wustl.edu
Staff Manager, EM Core Facility Fay and Carl Simons Center of Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
Did not see any response yet so I'll give it a try.
A good reference for SIMS is Wilson, R., Stevie, F. and Magee, C. (1989). Secondary ion mass spectrometry. New York: John wiley & Sons. ISBN 0-471-51945-6
Ion beam microscopy is a mode which is available in some, if not all (not sure) SIMS units. The distinction is made between depth profiles, side wall ion contributions and other effects. Large area ratios are typically required for probe mode to exclude secondary ions from the sidewalls when the beam is in the center of the crater. Alternatively, secondary ions are rejected outside the center of the crater "with an aperture for the ion microscope mode" (p. 1.5-1).
I haven't seen much other SIMS reference material either. Maybe it is a secret cult?
gary g.
At 04:57 PM 2/27/2002, you wrote:
} Hello All, } } I need some information. I hope I will get some responses from you. I was } wondering what the difference is between a SIMS Secondary Ion Mass } Spectrometer and an Ion Beam Electron Microscope. Please excuse my } ignorance, but I've tried looking on the web, and I haven't found the } explanation that I need. } } Any help would be appreciated. } } Thank you in Advance. } } Diane } } {mailto:millerd-at-coho.net} millerd-at-coho.net }
I'm looking for a book or some other resources on mineral analysis with EDS.
I have read a lot of information on the net about SEM / TEM and other instruments so I have a good theoretical knowledge about the process behind EDS but lacks the practical bit.
My background is a MS in physics, some chemistry and some geology. I have had some mineral samples analysed in a Russian lab and now I want to learn more about the practical side. How to interprete the results, how to prepare specimens, which problems could occur....
Is there a book or some other information source that you could recommend for me?
I will try to visit a lab for some hands on experience during the spring, but I would like to be well prepared so I could get the most out of the visit.
My final goal is to find a cheap used SEM with EDS to set up a small lab for mineral analyses.
JEOL5400LV probably is a first generation LV SEM. The weak point is its needle control valve on LV mode. It needs pay much attention to monitor the dial reading of pressure and manually adjust the pressure and make it stable. The new version LV SEMs have no manual control valve on the panel.
The LV SEM, with EDS attached on is very useful combination. You can do a lot of work on this system: EDS analysis on HV or LV, coated (on HV) or uncoated (on LV) samples, and LOW kv imaging (try 0.5-3.0 KV on HV mode) without coating. It even does not need to wear gloves at all. Get rid of restricted limitation and enjoy this powerful tool.
Zhiyu Wang, Sr. Engineer Maxtor Corp. Milpitas, CA USA
----- Original Message ----- } From: "Legendre Olivier" {Legendre-at-exchange.brgm.fr} To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, February 28, 2002 8:02 AM
My comment to add to the discussion having examined both LV and ESEM microscopes prior to purchasing an ESEM.
The strength of the LV SEM's is their ability to view SEM specimens without the need for coating. This they do very well and it is an enormously useful feature. Their ability to view wet specimens is limited by the fact that you cannot hold a particular RH (relative humidity) state for any length of time - as you can in the ESEM. In the LVSEM the water from a wet specimen would tend to leave the specimen rapidly. That said, we often use our ESEM in the 2 torr range for the examination of damp and/or greasy uncoated specimens where the maintanance of exact, or higher, water saturation levels is not critical.
Tony Bruton University of Natal South Africa
} } } "Richard Edelmann" {edelmare-at-muohio.edu} 02/28/02 07:02PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This is a beautiful low (lousy) vacuum SEM. Its sounds like the "restricted to study uncoated dry samples, preferably mineral" was put on the microscope by someone at the SEM Lab who didn't want any other samples/users using the scope.
I've often seen where the "Materials People" and the Geology people fobid messy unstable biological samples in their EM's in order to keep the scopes clean (particularly when dealing with analytical systems). (Now, where do the clays and rubber people fit in??)
Now, before you get someone upset you might find out the history of the "restricted to study uncoated dry samples, preferably mineral" and who paid for the scope (got the grant). But an EM is a terrible thing to waste.
On 28 Feb 2002, at 9:02, Legendre Olivier wrote:
} Hi all, } } I have been in charge recently of our SEM lab, in which there is a rather } idle JEOL5400LV (with a Kevex EDS) microscope. According to the person who } is in charge of running this machine, the capabilities of this SEM are } restricted to study uncoated dry samples, preferably mineral. } Does anybody have further experience on this machine (or an upgraded one?) } like imaging wet samples, etc.? } } Thank you for helping } } Olivier Legendre } BRGM } 3, avenue C. Guillemin } 45060 ORLEANS CEDEX 2 FRANCE } Tel: (33) 0 238 64 38 03 } Fax: (33) 0 238 64 37 11 } e-mail: o.legendre-at-brgm.fr } } } } } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
Hello, We met also the problem with the continuous one direction de-focusing with a heavy 16-slides Maerzhaeuser stage. We solved it in the software. Simply the drift was time calibrated. The time lapse experiment is running while the stage Z-drive compensates the drift according to the calibration. We are able to adapt this procedure almost to any motorised microscope.
-----Original Message----- } From: Andrew Ochalski [mailto:AOCHALSK-at-science.uottawa.ca] Sent: Thursday, February 28, 2002 5:24 PM To: Microscopy-at-sparc5.microscopy.com
Tony, I can't answer why it would be used, except that Cu-Be is much harder than Cu. The main danger from Cu-Be is if you grind it or burn it, but the grids are so small that grinding is fairly unlikely. As used in non-magnetic tools, etc., you're right. It's very safe.
Ken Converse owner Quality Images third party SEM service Delta, PA
Tony Garratt-Reed 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 not a "reply" to the post, but perhaps an extension of the } question. } } Why would Cu-Be grids be used? Cu-Be is an interesting and quite } widely-used class of alloys (see http://microstructure.copper.org), } but I don't see why it would be used for EM grids - unless, that is, } the grids we loosely call "copper" are in fact all made of Cu-Be? } } On the subject of the posting, I can't imagine that Cu-Be would be as } widely used as it is if it were as toxic as Be-metal. I'm sure the } alloying must drastically reduce the hazard, but this is not my area } of expertise. } } Tony } } } At 10:31 AM 2/28/2002 -0600, you 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 } } -----------------------------------------------------------------------. } } } } } } An individual new to EM wanted to learn from me how to formvar-carbon } } coat } } slotted grids. The person bought grids made of a Berylium-Copper } } alloy. I } } called the vendor and was warned about the danger of Berylium and it's } } vapor. I told the invidual to get copper grids for our use. Now in } } all my } } years in EM I have not worked with Berylium. Would someone out there } } send } } me some information on the hazards of Berylium? Also what } } applications are } } Berylium-Copper or Berylium only grids used for? Any information is } } appreciated, thanks. } } } } Tom Bargar } } EM Lab, UNMC } } tbargar-at-unmc.edu } } 402-559-7347 } } } } * * * * * * * * * * * * * * * * * * * * * * * * * * } * Anthony J. Garratt-Reed M.A., D.Phil. } * MIT, Room 13-1027 } * 77 Massachusetts Avenue } * Cambridge, MA 02139-4307 } * USA } * Phone: (617) 253-4622 } * Fax: (617) 258-6478 } * } } } }
You are correct in your 'secret cult' image, Gary. SIMS is largely a well-kept secret of both the geoscience and semiconductor communities, with the vast majority of materials scientists being unaware of its capabilities in trace element detection (including even hydrogen), sputter depth profiling, elemental imaging, isotope ratio age-dating and so forth. We here in Ottawa at a federal materials science-oriented lab have had a SIMS for nearly 15 years as a complement to our SEM, TEM and electron microprobe. Like our XPS and Auger, however, the SIMS is not used nearly as much as the EMs, but clients in the above two materials communities use it regularly. Let me note that the 'new kid on the block' in the SIMS community is something called a nanoSIMS, with a 50 nm beam (as opposed to the micron-ish beam of a regular SIMS) and simultaneous detection of up to 5 elements (much like a microprobe with WDX detectors). To date there are only two in North America, no surprise given the ~$2M+ US price tag.
However, I believe that Diane was referring to the differences between a SIMS and a focused ion beam (FIB) system, which is essentially a scanning ion microscope. In a FIB a much higher energy ion beam (30-50 keV as opposed to only several keV for a SIMS) is focused by electrostatic lens down to as little as 5 nm and scanned as in an SEM. The ion beam generates both secondary ions and secondary electrons, which can be captured to form the corresponding two types of images. Generally speaking, the resolution is intermediate between an LaB6 emitter SEM and a field emission (FE) SEM. The use of an electron flood gun permits good imaging of insulating materials. The most important difference between a SIMS and a FIB is that, while the former uses the scanned beam to sputter a crater for depth profiling, the latter uses its more energetic beam to accomplish in-situ 'micromachining'. Thus FIBs had their inception in the microelectronics community to conduct fine-scale repairs on devices. More recently, they have impacted seriously on general materials science via use of this micromachining capability to prepare parallel-sided thin sections for TEM in various ways. Finally, alas, FIBs have no analytical (EDXS) capability to date, save for a few models that combine both a ion beam column and an electron beam column (thus called dual beam FIBS), wherein an EDXS detector can be used in conjunction with the latter.
Attendees of M&M 2002 in Quebec City should check out the FIB session chaired by Lucille Gianuzzi and Mike Phaneuf. I guarantee you'll be impressed.
Tom
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada
} ---------- } From: Gary Gaugler } Sent: Thursday, February 28, 2002 6:20 PM } To: Diane G. Miller } Cc: MSA listserver } Subject: Re: SIMS } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Did not see any response yet so I'll give it a try. } } A good reference for SIMS is Wilson, R., Stevie, F. } and Magee, C. (1989). Secondary ion mass spectrometry. } New York: John wiley & Sons. ISBN 0-471-51945-6 } } Ion beam microscopy is a mode which is available in } some, if not all (not sure) SIMS units. The distinction } is made between depth profiles, side wall ion contributions } and other effects. Large area ratios are typically } required for probe mode to exclude secondary ions } from the sidewalls when the beam is in the center } of the crater. Alternatively, secondary ions are } rejected outside the center of the crater "with an } aperture for the ion microscope mode" (p. 1.5-1). } } I haven't seen much other SIMS reference material either. } Maybe it is a secret cult? } } gary g. } } } At 04:57 PM 2/27/2002, you wrote: } } } Hello All, } } } } I need some information. I hope I will get some responses from you. I } was } } wondering what the difference is between a SIMS Secondary Ion Mass } } Spectrometer and an Ion Beam Electron Microscope. Please excuse my } } ignorance, but I've tried looking on the web, and I haven't found the } } explanation that I need. } } } } Any help would be appreciated. } } } } Thank you in Advance. } } } } Diane } } } } {mailto:millerd-at-coho.net} millerd-at-coho.net } } } }
-----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-sparc5.microscopy.com] Sent: Thursday, February 28, 2002 9:41 AM To: NewSub-at-sparc5.microscopy.com
********************************************* This Email contains Important Information about the Microscopy Listserver. Please read it all then SAVE a copy for future reference. - Nestor **************************************** To: NewSub-at-MSA.Microscopy.Com } From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com}
As an occasional practitioner of the cult of SIMS, let me pass along an excellent web resource on the area.
http://www.simsworkshop.org
There is a good series of books on SIMS in the Springer Series in Chemical Physics, Secondary Ion Mass Spectrometry, ed. by Benninghoven, Colton, and Simons, but as these are conference proceedings more than strictly a reference book their value may initially not be as great as the one you suggested.
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Thursday, February 28, 2002 6:21 PM To: Diane G. Miller Cc: MSA listserver
Did not see any response yet so I'll give it a try.
A good reference for SIMS is Wilson, R., Stevie, F. and Magee, C. (1989). Secondary ion mass spectrometry. New York: John wiley & Sons. ISBN 0-471-51945-6
Ion beam microscopy is a mode which is available in some, if not all (not sure) SIMS units. The distinction is made between depth profiles, side wall ion contributions and other effects. Large area ratios are typically required for probe mode to exclude secondary ions from the sidewalls when the beam is in the center of the crater. Alternatively, secondary ions are rejected outside the center of the crater "with an aperture for the ion microscope mode" (p. 1.5-1).
I haven't seen much other SIMS reference material either. Maybe it is a secret cult?
gary g.
At 04:57 PM 2/27/2002, you wrote:
} Hello All, } } I need some information. I hope I will get some responses from you. I was } wondering what the difference is between a SIMS Secondary Ion Mass } Spectrometer and an Ion Beam Electron Microscope. Please excuse my } ignorance, but I've tried looking on the web, and I haven't found the } explanation that I need. } } Any help would be appreciated. } } Thank you in Advance. } } Diane } } {mailto:millerd-at-coho.net} millerd-at-coho.net }
Morning Goren (sorry for the missing um- [I'm just a] -lout?),
Now, I'm going to take you at your word and venture into an area which is NOT really mine yet. Briefly, I would suggest that you consult with Oxford Instruments for information on on a product they call "INCA Crystal" (no stock, no family and other companies such as Thermo NORAN also have such systems in the $50-$100k price range). These systems can be linked to the ICDD database for mineral identification and permit collection of diffracted X-rays phases within the specimen to determine both phase identification and crystal orientation as well as elemental mapping from EDS. I hope a practitioner responds, but here is the email of a vendor rep: Glenn Kinnear (Oxford): kinnear-at-ma.oxinst.com.
Hope this helps,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging West Chester University West Chester, Pennsylvania, USA, 19383 610-738-0437 fmonson-at-wcupa.edu
} ---------- } From: Göran Axelsson } Sent: Thursday, February 28, 2002 6:08 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM books on mineral analysis } } ------------------------------------------------------------------------ } 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! } } I'm looking for a book or some other resources on mineral analysis with } EDS. } } I have read a lot of information on the net about SEM / TEM and other } instruments so I have a good theoretical knowledge about the process } behind EDS but lacks the practical bit. } } My background is a MS in physics, some chemistry and some geology. } I have had some mineral samples analysed in a Russian lab and now I } want to learn more about the practical side. } How to interprete the results, how to prepare specimens, which problems } could occur.... } } Is there a book or some other information source that you could recommend } for me? } } I will try to visit a lab for some hands on experience during the spring, } but I } would like to be well prepared so I could get the most out of the visit. } } My final goal is to find a cheap used SEM with EDS to set up a small lab } for } mineral analyses. } } Regards, Göran Axelsson, Sweden } } } }
62 glorious pages that advertise the dangers of Be from the DOE, probably its largest user!
URL:http://tis-nt.eh.doe.gov/be/berule.pdf (WARNING! You should have the Adobe Acrobat Reader before trying this URL)
Hope this helps. You can also try the ToxNet on National Library of Medicine gateway to MEDLINE: http://gateway.nlm.nih.gov/gw/Cmd.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging West Chester University West Chester, Pennsylvania, USA, 19383 610-738-0437 fmonson-at-wcupa.edu
} ---------- } From: "tbargar-at-unmc.edu"-at-sparc5.microscopy.com } Sent: Thursday, February 28, 2002 11:31 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Need Info. about Berylium and Berylium-Copper slotted grids } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } An individual new to EM wanted to learn from me how to formvar-carbon coat } slotted grids. The person bought grids made of a Berylium-Copper alloy. } I } called the vendor and was warned about the danger of Berylium and it's } vapor. I told the invidual to get copper grids for our use. Now in all } my } years in EM I have not worked with Berylium. Would someone out there send } me some information on the hazards of Berylium? Also what applications } are } Berylium-Copper or Berylium only grids used for? Any information is } appreciated, thanks. } } Tom Bargar } EM Lab, UNMC } tbargar-at-unmc.edu } 402-559-7347 } } }
We have a JEOL JSM5400LV that, as far as I now, was one of the very first of this model. It is true that it is not a true environmental SEM, if you define that as a SEM in which you can study wet samples.
However, we have great use for the low vacuum mode of the instrument for looking at insulating samples without coating. The insulating samples are f.x. polymers, corrosion products and various dusts and contaminations.
The main limitation compared to an environmental SEM is that we are not able to use the secondaary electron detector in LV mode. Instead we use the backscatter detector that does quite a good job of "topographic" imaging as long as we do not need high magnification (~} 5000x).
Yours sincerely,
Henning Sund Sørensen Materials- and Process Consultant
Danfoss A/S Central Service Technology Centre Nordborgvej 81, L7-S40 6430 Nordborg Denmark
-----Original Message----- } From: Legendre Olivier [mailto:Legendre-at-exchange.brgm.fr] Sent: 28. februar 2002 09:02 To: 'Microscopy-at-MSA.Microscopy.Com'
Hi all,
I have been in charge recently of our SEM lab, in which there is a rather idle JEOL5400LV (with a Kevex EDS) microscope. According to the person who is in charge of running this machine, the capabilities of this SEM are restricted to study uncoated dry samples, preferably mineral. Does anybody have further experience on this machine (or an upgraded one?) like imaging wet samples, etc.?
About 8-10 years ago we had upgraded our Kevex Sesame/Microspec system by replacing the Sesame system with a 386 PC and card/software we purchased from Microspec. We don't use the system often but when we need the better energy resolution because of peak overlap in the XEDS it is the only way to go. We had dealt with Steve Carr (really streching my memory) at Microspec back then. Microspec was taken over by Oxford Instruments, I believe. This is a link to the appropriate location in their web site; http://www.oxford-instruments.com/ANLPDP177.htm
There are other third party systems available as well; http://www.advancedmicrobeam.com/micro3wd.htm
I have no financial interest in any of these companies.
At the moment we have no plans to excess our Microspec so sorry no spare boards. Within the last two years we did have a problem with the high voltage to the detector and we were able to trace the problem to a bad capacitor on the high voltage board. Otherwise the detector has performed well.
} } } Greetings all, } } I would be interested to hear from anyone who is still using a Kevex } Sesame24/Microspec 2A WDS system. If you have one that is retired, I would } especially have an interest in the possibility of obtaining spare cards. } Mine is working but have no spares... } } Another question about the Sesame... I no longer have the Kevex 8000/Delta } EDS to which the WDS sent data for quantitation. Has anyone mated the } Sesame to a different EDS/software package to do quantitation? } } Thanks, Woody } -------------- } Woody White } McDermott Technology, inc } nwwhite-at-mcdermott.com
-- David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
The book I have used for years is called "SEM Petrology Atlas" by Joann E. Welton. It was published in 1984 by the American Association of Petroleum Geologists (Tulsa, OK 74101) as part of the Methods in Exploration Series. Not sure if it still available, but the ISBN # is 0-89181-653-4.
It contains images and EDS spectra of a variety of minerals - silicates, carbonates, phosphates, halides, sulfides, sulfates, and oxides.
Hope this helps, Lou Ross
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211-5120 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu web: www.biotech.missouri.edu/emc
_SEM Petrology Atlas_ by Joann E. Welton, published by The American Association of Petroleum Geologists, Tulsa, OK 74101 USA; Copyright 1984, ISBN 0-89181-653-4.
It contains SEM micrographs and EDS spectra of various minerals associated with oil and gas exploration. I am a metallurgist, not a geologist, and this book has saved my... day... a number of times.
Regards, Andrew T. Werner Chief Metallurgist Shaped Charge Research - Metallurgy Laboratory Schlumberger Reservoir Completions Technology Center 14910 Airline Road, Rosharon, TX 77583-1590 Voice (281) 285-5272 Fax (281) 285-5273
"We shoot the hippopotamus with bullets made of platinum 'cause if we used the leaden ones his hide would surely flatten 'em" - Author Unknown
At 12:08 AM 3/1/2002 +0100, Göran Axelsson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Could be she was refering to SIMS and FIB. Based on my experience with FEI FIB (old model 611), it has poor imaging resolution. Their newer models, like the 830, are as you say, dual beam--ion and electron. The electron beam is used for imaging while the ion beam is used for micro machining, etc. FIBs are great for making microcircuit cross sections and changing runners on the planar area of a chip.
Supposedly, the FEI dual beam FIB will accept a SIMS "detector." So it would do a whole bunch of good tasks. Maybe there is only one spare port. Either a SIMS detector or x-ray detector could be fitted.
gary g.
At 05:05 AM 3/1/2002, you wrote: } You are correct in your 'secret cult' image, Gary. SIMS is largely a } well-kept secret of both the geoscience and semiconductor communities, with } the vast majority of materials scientists being unaware of its capabilities } in trace element detection (including even hydrogen), sputter depth } profiling, elemental imaging, isotope ratio age-dating and so forth. We } here in Ottawa at a federal materials science-oriented lab have had a SIMS } for nearly 15 years as a complement to our SEM, TEM and electron microprobe. } Like our XPS and Auger, however, the SIMS is not used nearly as much as the } EMs, but clients in the above two materials communities use it regularly. } Let me note that the 'new kid on the block' in the SIMS community is } something called a nanoSIMS, with a 50 nm beam (as opposed to the micron-ish } beam of a regular SIMS) and simultaneous detection of up to 5 elements (much } like a microprobe with WDX detectors). To date there are only two in North } America, no surprise given the ~$2M+ US price tag. } } However, I believe that Diane was referring to the differences between a } SIMS and a focused ion beam (FIB) system, which is essentially a scanning } ion microscope. In a FIB a much higher energy ion beam (30-50 keV as } opposed to only several keV for a SIMS) is focused by electrostatic lens } down to as little as 5 nm and scanned as in an SEM. The ion beam generates } both secondary ions and secondary electrons, which can be captured to form } the corresponding two types of images. Generally speaking, the resolution } is intermediate between an LaB6 emitter SEM and a field emission (FE) SEM. } The use of an electron flood gun permits good imaging of insulating } materials. The most important difference between a SIMS and a FIB is that, } while the former uses the scanned beam to sputter a crater for depth } profiling, the latter uses its more energetic beam to accomplish in-situ } 'micromachining'. Thus FIBs had their inception in the microelectronics } community to conduct fine-scale repairs on devices. More recently, they } have impacted seriously on general materials science via use of this } micromachining capability to prepare parallel-sided thin sections for TEM in } various ways. Finally, alas, FIBs have no analytical (EDXS) capability to } date, save for a few models that combine both a ion beam column and an } electron beam column (thus called dual beam FIBS), wherein an EDXS detector } can be used in conjunction with the latter. } } Attendees of M&M 2002 in Quebec City should check out the FIB session } chaired by Lucille Gianuzzi and Mike Phaneuf. I guarantee you'll be } impressed. } } Tom } } Tom Malis } Group Leader - Characterization } Materials Technology Laboratory } Natural Resources Canada (Govt. of Canada) } 568 Booth St., Ottawa, Canada } } } ---------- } } From: Gary Gaugler } } Sent: Thursday, February 28, 2002 6:20 PM } } To: Diane G. Miller } } Cc: MSA listserver } } Subject: Re: SIMS } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } Did not see any response yet so I'll give it a try. } } } } A good reference for SIMS is Wilson, R., Stevie, F. } } and Magee, C. (1989). Secondary ion mass spectrometry. } } New York: John wiley & Sons. ISBN 0-471-51945-6 } } } } Ion beam microscopy is a mode which is available in } } some, if not all (not sure) SIMS units. The distinction } } is made between depth profiles, side wall ion contributions } } and other effects. Large area ratios are typically } } required for probe mode to exclude secondary ions } } from the sidewalls when the beam is in the center } } of the crater. Alternatively, secondary ions are } } rejected outside the center of the crater "with an } } aperture for the ion microscope mode" (p. 1.5-1). } } } } I haven't seen much other SIMS reference material either. } } Maybe it is a secret cult? } } } } gary g. } } } } } } At 04:57 PM 2/27/2002, you wrote: } } } } } Hello All, } } } } } } I need some information. I hope I will get some responses from you. I } } was } } } wondering what the difference is between a SIMS Secondary Ion Mass } } } Spectrometer and an Ion Beam Electron Microscope. Please excuse my } } } ignorance, but I've tried looking on the web, and I haven't found the } } } explanation that I need. } } } } } } Any help would be appreciated. } } } } } } Thank you in Advance. } } } } } } Diane } } } } } } {mailto:millerd-at-coho.net} millerd-at-coho.net } } } } } } }
Is anyone interested in purchasing a Bio-Rad MRC 600 confocal microscope system? Included are: 1. Krypton-Argon Laser (low hours) and power source exciting at 488 568 and 655 nm. 2. Nikon optiphot upright epifluorescence microscope with 10 20 and 60x Plan Apo objective lens', mercury arc lamp and power source. 3. Fine focus control stepping motor. 4. Compaq Desktop Pro pentium pc platform wtih Comos and Som software, and two color monitors. 5. Mitsubishi Dye-sublimation printer.
This system is operational and available for $10,000.000. We will ship, set-up and train the buyer. Please contact Michael Delannoy at (410) 955-1365 or via e-mail.
Thank you, Michael Delannoy Assist. Direct. Microscopy Facility JHMI
P.S. Does anyone have the confocal listserver address?
Since I have many years of SIMS experience, I responded to the recent question on this subject directly. I would like to let the group know that there is a SIMS list at sims-at-sims.arl.army.mil. There is also a Web site at http://www.simsworkshop.org/default.nclk with lots of information. SIMS is an expensive technique to own so the user community is small compared to SEM but if you are looking for low level elemental information its the best technique for the job.
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
I can only imagine Cu-Be grids are used for there superior stiffness compared to Cu grids. The amount of Be in these alloys is typically 1-2 wt%. The alloy is used to manufacture non-magnetic tools, springs (it can sustain a greater deflection before permanently deforming than spring steel up to 200degC) and in the chemical and electrical industry fields where there is a risk of explosion as the alloy is non-sparking.
Be grids were available for a time although I do not think they are now. The low atomic number meant that there was no stray x-rays detected from the grid material. Be is still extensively used in Materials Science TEM specimen holders (low background) for X-ray analysis because of this.
The dangers of Be and its alloys and compounds can be found on the SIRI MSDS website:-
siri.org/msds/index.php
Certain precautions are necessary when using Be. Any ingestion of the metal or its compounds should be avoided. Gloves should be worn at all times when handling Be or Be containing alloys. There is significantly less risk of symptoms from Cu-Be alloys although the effects of exposure to Be is cumulative (and delayed). Be dust is particularly dangerous it should not be inhaled or ingested, it is a carcinogen causes beryllium disease, a particularly chronic lung disease, and is an explosion hazard in air in high concentrations. Be containing parts should not be machined or filed. Companies machining parts out of Be do so in carefully monitored glove boxes with special filters to avoid the dust being dispersed in the atmosphere.
For occasional exposure to Be typical of life in an EM lab, as long as people are made aware of the dangers and use gloves, do not swallow the parts (!) and certainly do not take a file to them (!!) they are safe. Care must be taken however not to lose Be parts and if they are to be disposed of, they must not just be thrown away!
Regards
Alan
At 04:06 PM 2/28/2002 -0500, Tony Garratt-Reed wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Anyone out there with experience using a cooled digital camera to capture the luminescence of luciferase? We are looking at a macroscopic specimen expressing a luciferase tag using a 20x NA = 0.7 objective and a Sensys camera. I am assuming a don't need to have a filter in the path since the sample is only source of light. Does this seem practical? SO far we have had no luck. Any tips or tricks would be greatly appreciated. -- 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)
Tom; Beryllium is very potent at hardening copper without significantly reducing it's electrical conductivity. It is commonly used for springs and contacts in electrical switches, and is present at such a low concentration (~0.5%) that it is not considered a hazard. It also does not introduce any background that would intrude on EDX analysis. We use Be Cu slotted grids as substrates for 20 micron thick silicon slivers that will be milled in a focused ion beam tool. The substrate should resist bending because that would lead to fracture of the silicon. Unalloyed copper would bend too easily and would not be suitable.
John Mardinly Intel
-----Original Message----- } From: "tbargar%unmc.edu"-at-sparc5.microscopy.com [mailto:"tbargar%unmc.edu"-at-sparc5.microscopy.com] Sent: Thursday, February 28, 2002 8:31 AM To: Microscopy-at-sparc5.microscopy.com
An individual new to EM wanted to learn from me how to formvar-carbon coat slotted grids. The person bought grids made of a Berylium-Copper alloy. I called the vendor and was warned about the danger of Berylium and it's vapor. I told the invidual to get copper grids for our use. Now in all my years in EM I have not worked with Berylium. Would someone out there send me some information on the hazards of Berylium? Also what applications are Berylium-Copper or Berylium only grids used for? Any information is appreciated, thanks.
Tom Bargar EM Lab, UNMC tbargar-at-unmc.edu 402-559-7347
Dear Tom, Berylium-copper is an expensive alloy that allows a higher strength and hardness than most copper alloys while keeping the non-magnetic and non-sparking properties of copper. I used to have a set of tools for my ETEC SEM made of Be-Cu. The Be is less than 2.0 weight percent of the alloy and it is very tightly bound in the copper, so I don't think there is any danger of Be exposure on normal use or mild heating. The main danger of Be is the inhalation of BeO in the dust if Be is ground. At 10:31 AM 2/28/02 -0600, you wrote: } } An individual new to EM wanted to learn from me how to formvar-carbon coat } slotted grids. The person bought grids made of a Berylium-Copper alloy. I } called the vendor and was warned about the danger of Berylium and it's } vapor. I told the invidual to get copper grids for our use. Now in all my } years in EM I have not worked with Berylium. Would someone out there send } me some information on the hazards of Berylium? Also what applications are } Berylium-Copper or Berylium only grids used for? Any information is } appreciated, thanks. } } Tom Bargar } EM Lab, UNMC } tbargar-at-unmc.edu } 402-559-7347 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-interchange.ubc.ca
I would like to solicit any info about automated grid stainers. I have been always staining by hand. Need to know pros and cons about the different ones that are in use. Any information, such as; initial cost, size, reproducibility, quality, open/closed system, reagents utilized, waste disposal, and etc., etc., will be appreciated.
Thank you TEM netters,
Donald G. Awbrey, HT(ASCP) QIHC Electron Microscopy / Image Analysis 817-878-5647 donaldawbrey-at-texashealth.org
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Anyone know where I can buy some Euparal? It's a mounting medium for light microscopy, often used by entomologists et al. WWW search and other tries here have turned up nil.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
on 2/28/02 11:31 AM, "tbargar-at-unmc.edu"-at-sparc5.microscopy.com at "tbargar-at-unmc.edu"-at-sparc5.microscopy.com wrote:
} Also what applications are } Berylium-Copper or Berylium only grids used for? Any information is } appreciated, thanks. } Dear Tom, Be grids are used for EDS, since their lower Z means that they scatter fewer electrons and produce fewer brehmsstrahlung x-rays than other materials, so they give a very low background count for x-ray analysis. Also, the only characteristic x-rays they produce are very low energy and do not interfere with most analyses. Yours, Bill Tivol
on 2/28/02 4:06 PM, Tony Garratt-Reed at tonygr-at-mit.edu wrote: } } This is not a "reply" to the post, but perhaps an extension of the question. } } Why would Cu-Be grids be used? Cu-Be is an interesting and quite } widely-used class of alloys (see http://microstructure.copper.org), but I } don't see why it would be used for EM grids - unless, that is, the grids we } loosely call "copper" are in fact all made of Cu-Be? } } On the subject of the posting, I can't imagine that Cu-Be would be as } widely used as it is if it were as toxic as Be-metal. I'm sure the } alloying must drastically reduce the hazard, but this is not my area of } expertise. } } Tony } } } } } An individual new to EM wanted to learn from me how to formvar-carbon coat } } slotted grids. The person bought grids made of a Berylium-Copper alloy. I } } called the vendor and was warned about the danger of Berylium and it's } } vapor. I told the invidual to get copper grids for our use. Now in all my } } years in EM I have not worked with Berylium. Would someone out there send } } me some information on the hazards of Berylium? Also what applications are } } Berylium-Copper or Berylium only grids used for? Any information is } } appreciated, thanks. } } } } Tom Bargar
Dear Tony & Tom, Cu-Be is used to make non-magnetic tools, and I'm sure it is used in preference to copper or bronze because of its strength and toughness. I suspect that the same qualities would make the Cu-Be grids more durable. Be metal is toxic in the Be++ form, so the oxide or other compounds are more toxic than the metal itself. The reason one should be careful handling the metal is that it can dissolve in the fluid in breaks in the skin, etc., and be introduced into the blood in the toxic form. Apparently, this does not happen with the Cu-Be alloy--at least, I've never seen warnings about using the tools, which often are used by people with cuts or abrasions on the skin. I'd be interested if anyone knows differently. Yours, Bill Tivol
Ken; When I worked at Lockheed, all beryllium alloys that were cut, ground, or polished had to be prepared in a special lab with numerous special safety precautions. Beryllium copper alloys (0.5%) were never subject to these safety rules, and could be cut, ground and polished without special precautions in any lab.
John Mardinly Intel
-----Original Message----- } From: Ken Converse [mailto:qualityimages-at-netrax.net] Sent: Friday, March 01, 2002 3:38 AM To: Tony Garratt-Reed; MSA, listserver
Tony, I can't answer why it would be used, except that Cu-Be is much harder than Cu. The main danger from Cu-Be is if you grind it or burn it, but the grids are so small that grinding is fairly unlikely. As used in non-magnetic tools, etc., you're right. It's very safe.
Ken Converse owner Quality Images third party SEM service Delta, PA
Tony Garratt-Reed 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 not a "reply" to the post, but perhaps an extension of the } question. } } Why would Cu-Be grids be used? Cu-Be is an interesting and quite } widely-used class of alloys (see http://microstructure.copper.org), } but I don't see why it would be used for EM grids - unless, that is, } the grids we loosely call "copper" are in fact all made of Cu-Be? } } On the subject of the posting, I can't imagine that Cu-Be would be as } widely used as it is if it were as toxic as Be-metal. I'm sure the } alloying must drastically reduce the hazard, but this is not my area } of expertise. } } Tony } } } At 10:31 AM 2/28/2002 -0600, you 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 } } -----------------------------------------------------------------------. } } } } } } An individual new to EM wanted to learn from me how to formvar-carbon } } coat } } slotted grids. The person bought grids made of a Berylium-Copper } } alloy. I } } called the vendor and was warned about the danger of Berylium and it's } } vapor. I told the invidual to get copper grids for our use. Now in } } all my } } years in EM I have not worked with Berylium. Would someone out there } } send } } me some information on the hazards of Berylium? Also what } } applications are } } Berylium-Copper or Berylium only grids used for? Any information is } } appreciated, thanks. } } } } Tom Bargar } } EM Lab, UNMC } } tbargar-at-unmc.edu } } 402-559-7347 } } } } * * * * * * * * * * * * * * * * * * * * * * * * * * } * Anthony J. Garratt-Reed M.A., D.Phil. } * MIT, Room 13-1027 } * 77 Massachusetts Avenue } * Cambridge, MA 02139-4307 } * USA } * Phone: (617) 253-4622 } * Fax: (617) 258-6478 } * } } } }
A clarification is probably in order. The INCA Crystal software actually works with scattered electrons (rather than x-rays) to determine the crystallography of the phase.
EDS is good for identifying most minerals, especially given some background of the likely candidates. But there are many cases of ambiguity until crystallographic information is available.
I know that EDS systems are available out there for around $60K. I am practically certain that would not include the INCA Crystal hardware and software. I would expect that to nearly double the cost of the system, but I haven't priced one yet.
Warren
At 08:39 AM 3/1/02 -0500, you wrote:
} Morning Goren (sorry for the missing um- [I'm just a] -lout?), } } Now, I'm going to take you at your word and venture into an area } which is NOT really mine yet. } Briefly, I would suggest that you consult with Oxford Instruments } for information on on a product they call "INCA Crystal" (no stock, no } family and other companies such as Thermo NORAN also have such systems in } the $50-$100k price range). These systems can be linked to the ICDD } database for mineral identification and permit collection of diffracted } X-rays phases within the specimen to determine both phase identification and } crystal orientation as well as elemental mapping from EDS. } I hope a practitioner responds, but here is the email of a vendor } rep: Glenn Kinnear (Oxford): kinnear-at-ma.oxinst.com. } } Hope this helps, } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging } West Chester University } West Chester, Pennsylvania, USA, 19383 } 610-738-0437 } fmonson-at-wcupa.edu } } } ---------- } } From: Göran Axelsson } } Sent: Thursday, February 28, 2002 6:08 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: SEM books on mineral analysis } } } } Hello! } } } } I'm looking for a book or some other resources on mineral analysis with } } EDS. } } } } I have read a lot of information on the net about SEM / TEM and other } } instruments so I have a good theoretical knowledge about the process } } behind EDS but lacks the practical bit. } } } } My background is a MS in physics, some chemistry and some geology. } } I have had some mineral samples analysed in a Russian lab and now I } } want to learn more about the practical side. } } How to interprete the results, how to prepare specimens, which problems } } could occur.... } } } } Is there a book or some other information source that you could recommend } } for me? } } } } I will try to visit a lab for some hands on experience during the spring, } } but I } } would like to be well prepared so I could get the most out of the visit. } } } } My final goal is to find a cheap used SEM with EDS to set up a small lab } } for } } mineral analyses. } } } } Regards, Göran Axelsson, Sweden
Director, Core Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
Email: avklaus-at-amnh.org Tel: 212-769-5977
On Fri, 1 Mar 2002, 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: } } Anyone know where I can buy some Euparal? It's a mounting medium for light } microscopy, often used by entomologists et al. WWW search and other tries } here have turned up nil. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dan-at-isaacson.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, March 1, 2002 at 17:27:04 ---------------------------------------------------------------------------
Email: dan-at-isaacson.net Name: Dan Isaacson
Organization: Seattle Central Community College
Education: Undergraduate College
Location: Seattle, Washington
Question: I live in Seattle and I was wondering what would be the best place/microscope/technique to observe cellular mitosis. More specifically I'm looking to observe the mitoic spindles through interphase, prophase, prometaphase, metaphase, anaphase, and telophase. Considering the size of spindles and the microtubule fibers that connect to the chromosomes it may be very difficult to observe. But any help you can give me would be greatly appriciated.
We need a cooled chamber large enough to hold a multiwell plate, 3"X6" (approximately), able to hold the temperature at 4 degrees C. This chamber will be used with a stereo zoom microscope at relatively high power and we must be able to pipette the multiwell plate through a port in the chamber. If you have any information on manufacturers of a device like this, or know of someone who has built one, please share the information with us. We would like to avoid reinventing the wheel on this project and hope somebody has done this already. We are are about to start fabricating one because we haven't found a source for one.
We are able to make any modifications needed to make a chamber that is close to what we need. This research previously has been done with the microscope and researcher inside a large refrigerater! Burr...
Thanks for your help. Dave Burton University of Washington, Seattle
The "SEM Petrology Atlas" by Joann Welton is out of print. I tried to find a copy several months ago but was unable to. I checked Amazon.com today and there is one copy available from a used book dealer for US $371. A bit pricy but if you plan on looking at a lot of rocks, especially sedimentary rocks, this is an excellent reference. You might want to search Amazon.com to find some more recent (and less expensive) books.
Most of my work is X-ray diffraction on rocks but I do some SEM work. The sample prep is pretty simple. If you have any specific questions, feel free to contact me off-list.
Sincerely, James Talbot
K/T GeoServices, Inc. Bulk and Clay Mineralogy by X-ray Diffraction (940) 597-9076 web site: http://www.ktgeo.com/
Lou Ross 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 Goran, } } The book I have used for years is called "SEM Petrology Atlas" by Joann E. } Welton. It was published in 1984 by the American Association of Petroleum } Geologists (Tulsa, OK 74101) as part of the Methods in Exploration Series. } Not sure if it still available, but the ISBN # is 0-89181-653-4. } } It contains images and EDS spectra of a variety of minerals - silicates, } carbonates, phosphates, halides, sulfides, sulfates, and oxides. } } Hope this helps, } Lou Ross } } } ------------------------------------------------------------------------ } } 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! } } } } I'm looking for a book or some other resources on mineral analysis with } } EDS. } } } } I have read a lot of information on the net about SEM / TEM and other } } instruments so I have a good theoretical knowledge about the process } } behind EDS but lacks the practical bit. } } } } My background is a MS in physics, some chemistry and some geology. } } I have had some mineral samples analysed in a Russian lab and now I } } want to learn more about the practical side. } } How to interprete the results, how to prepare specimens, which problems } } could occur.... } } } } Is there a book or some other information source that you could } } recommend } } for me? } } } } I will try to visit a lab for some hands on experience during the } } spring, but I } } would like to be well prepared so I could get the most out of the visit. } } } } My final goal is to find a cheap used SEM with EDS to set up a small lab } } for } } mineral analyses. } } } } Regards, Göran Axelsson, Sweden } } Senior Electron Microscope Specialist } Electron Microscopy Core Facility } W136 Veterinary Medicine } University of Missouri } Columbia, MO 65211-5120 } (573) 882-4777, fax 884=5414 } email: rosslm-at-missouri.edu } web: www.biotech.missouri.edu/emc
I have questions pertaining to practical applications for this BSE/CL detector. Would any of you who have practical experience please contact me directly.
genuinely ... michael shaffer :o) Avalon Peninsula, Newfoundland www.micro-investigations.com
I have questions pertaining to practical applications for this BSE/CL detector. Would any of you who have practical experience please contact me directly.
genuinely ... michael shaffer :o) Avalon Peninsula, Newfoundland www.micro-investigations.com
Many years ago (1986) we were attempting to clean Formvar coating and epoxy resin sections from slotted beryllium grids. Since, for safety reasons, acid could not be used to clean beryllium, we were following the suppplier's suggestion to use either acetone or ethanol. When an initial wash in acetone did not do a satisfactory job, the grids were placed in 100% ethanol. Shortly thereafter, examination of the grids under a dissecting microscope revealed distinct signs of corrosion: holes appeared in one grid, and pieces broke from the edges of two others.
We immediately contacted the supplier, who immediately called the manufacturer, who was of the opinion that the formaldehyde in Formvar, in conjunction with an organic solvent, could indeed initiate corrosive action on the beryllium. Proper disposal consisted of pouring the grids and the ethanol solution onto filter paper in a funnel; the wet filter paper with the grids on it was placed in a plastic bag (to prevent drying out and subsequent release of dust) for pick-up by the biological safaety office, and the liquid was poured down the sink.
At that time beryllium itself was not considered nearly as toxic as it had once been, but beryllium salts were still very bad news. As a result of our experience, the supplier (Ted Pella) decided to suggest to all customers that NO attempt be made to clean beryllium grids, but that they be discarded in a safe manner after use.
Alan Nicholls wrote: ========================================== I can only imagine Cu-Be grids are used for there superior stiffness compared to Cu grids. The amount of Be in these alloys is typically 1-2 wt%. The alloy is used to manufacture non-magnetic tools, springs (it can sustain a greater deflection before permanently deforming than spring steel up to 200degC) and in the chemical and electrical industry fields where there is a risk of explosion as the alloy is non-sparking.
Be grids were available for a time although I do not think they are now. The low atomic number meant that there was no stray x-rays detected from the grid material. Be is still extensively used in Materials Science TEM specimen holders (low background) for X-ray analysis because of this. =========================================== We might be comparing apples with oranges here.
What is referred to as "copper/beryllium alloy" indeed is mainly copper with a little bit of beryllium added. I have worked in the M&M field for more years than I will admit and I have never heard of anyone making either grids or planchettes out of that alloy. After all, it would be self-defeating and would serve no useful purpose in EM.
There are alloys of Be that are 98 and 99% Be and the non-Be content can be Cu and/or other impurities. Over the years, grids and planchettes have been offered of the 98 and 99% purity alloys, and they were somewhat cheaper than the higher purity 99.8% Be that have been offered now for some years by SPI Supplies. If you have grids where no statement of purity was given, we believe that you can probably assume they were 98% or possibly 99% but not the 99.8% level.
At one time, and they might still be offered, were essentially copper grids that had a sputter coated layer of Be. I think the theory was that the Be coating approach could lead to a lower priced product that could do the same thing. We always had the idea that they did not work very well in terms of keeping Cu lines out of the EDS spectra. That might not be the experience of others, however. We also heard reports of the Be layer flaking off as the grid was flexed, which could lead to an inhalation hazard.
A full range of 99.8% purity Be grids is described on URL http://www.2spi.com/catalog/grids/beryl.html
They are very much still available off-the-shelf from SPI Supplies. For those who are in institutions that have banned Be in any form, the above URL mentions and provides links to product alternatives.
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 ######################## ============================================
Joanne Whallon wrote: ================================================================= Many years ago (1986) we were attempting to clean Formvar coating and epoxy resin sections from slotted beryllium grids. Since, for safety reasons, acid could not be used to clean beryllium, we were following the supplier's suggestion to use either acetone or ethanol. When an initial wash in acetone did not do a satisfactory job, the grids were placed in 100% ethanol . Shortly thereafter, examination of the grids under a dissecting microscope revealed distinct signs of corrosion: holes appeared in one grid, and pieces broke from the edges of two others.
We immediately contacted the supplier, who immediately called the manufacturer, who was of the opinion that the formaldehyde in Formvar, in conjunction with an organic solvent, could indeed initiate corrosive action on the beryllium. Proper disposal consisted of pouring the grids and the ethanol solution onto filter paper in a funnel; the wet filter paper with the grids on it was placed in a plastic bag (to prevent drying out and subsequent release of dust) for pick-up by the biological safety office, and the liquid was poured down the sink.
At that time beryllium itself was not considered nearly as toxic as it had once been, but beryllium salts were still very bad news. As a result of our experience, the supplier (Ted Pella) decided to suggest to all customers that NO attempt be made to clean beryllium grids, but that they be discarded in a safe manner after use. ========================================================== A minute or less in an oxygen plasma, such as what is produced in the SPI Plasma Prep II plasma etcher will "etch" away anything organic in the way of a TEM thickness section. I am talking about seconds. Because of the isotropic nature of the etching process in the Plasma Prep II (as opposed to anisotropic etchers), both sides seemed to get cleaned, although if it was me, I would try to put the section side up.
One never knows for sure of course, but the etching rate with oxygen on Beryllium metal would be essentially zero when compared to anything organic.
And while it might be a digression from the main thread of the topic, let me point out that the hazard is an inhalation hazard, namely beryllium oxide (BeO). Beryllium metal is about as inert and innocuous as anything you can find. A dangerous situation does not arise just have having the grid (or planchette) sitting there, or for that matter, putting the grid in the TEM (or SEM) but from the processing of the item. For example, those who might try "repolishing" a Be planchette take on an especially high risk because if their polishing table is allowed to dry out, there could result a dry powder that has become BeO and it would indeed become a hazard if the polishing media was dislodged and the entrapped particulates became air borne. I have not heard of an analogous situation with regard to the cleaning, mechanically, the grids.
If one was concerned about beryllium in any form exiting the mechanical pump being used with a plasma etcher, and if a standard oil mist filter was deemed to not be enough protection, then the pump can be vented directly to the laboratory's fume exhaust system (or operated inside of the fume hood).
Finally, contrary to conventional wisdom in M&M land, many of these products , including Be grids are not all the same, especially with regard to non- breyllium content, such as copper. Additional information can be found on URL http://www.2spi.com/catalog/grids/note.html
The chemical resistance of beryllium alloyed with copper will be considerably less, and therefore explaining the pitting, than the higher purity preferred by those who use these kinds of grids on a regular basis. The point is that your experience with the etching suggests the non- beryllium content of the grids was high and such experience could not be expected to be reproduced by someone using the higher purity (and more expensive beryllium foil) that is used in the making of the higher Be content grids.
Disclaimer: SPI Supplies is a supplier of high purity beryllium grids so we would have a vested interest in seeing more people use SPI Supplies® brand of beryllium grids than grids offered by others.
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 ######################## ============================================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lisa.monaco-at-msfc.nasa.gov) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, March 1, 2002 at 13:27:42 ---------------------------------------------------------------------------
Email: lisa.monaco-at-msfc.nasa.gov Name: lisa monaco
Organization: MSFC
Education: Graduate College
Location: Hunstville Alabama
Question: What would be the some of the best optical techniques for high resolution imaging of transparent cubic crystals in solution.
The are 20 microns on edge, at high resolution (i.e., we want to be able to make measurements on crystal dimensions within about 5-10 microns) and they are in transparent solution from which they grew. In some cases, just a phase separation (for example SCN crystals out of SCN solution)