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From: Asbjrn Skogstad :      asbjorsk-at-stami.no
Date: Tue, 01 Feb 2000 12:17:19 +0100
Subject: LM,fluorescence - aggregation of bacteria

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-Obviously an advantage for many bacteria species, but a nightmare for
the microscopist who wants to count
them. For several reasons we want to split the aggregates of bacteria
into single cells before counting.
We have tried mild detergent treatment and ultrasound though with
limited success. The samples are initially
taken, as filter samples in working atmospheres were bacteria could be
airborne, e.g. farm work. Afterwards
the bacteria are washed off the filter, resuspended, stained (AO),
refiltered on black pc-filter, mounted and
counted. Any suggestions for a treatment, which will de-aggregate the
bacteria, are most welcome.

Asbjorn Skogstad
National Inst. of Occup. Health,
Oslo, Norway
asbjorn.skogstad-at-stami.no





From: Tony Bruton :      bruton-at-EMU.UNP.AC.ZA
Date: 1/31/00 11:06 AM
Subject: Sandbox Squabble-Weather

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Chuck 'an all

That's why I tried to bring in a useful element to it by mentioning, a little tongue in cheek, my humidity vs formvar films question. But thanks to the silly weather thread I had some useful and interesting responses. May I be forgiven for saying that there's a silver lining..............

Tony Bruton
University of Natal
South Africa

} } } "Chuck Butterick" {cbutte-at-ameripol.com} 02/01/00 04:06AM } } }
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The whole weather thing has been pretty silly.

Chuck Butterick
Engineered Carbons, Inc.


______________________________ Reply Separator _________________________________


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Hi,

Are we like kids sitting in a sandbox and squabbling? Yes weather, no
weather! He said it first! She started it! I don't want weather on the
listserver. I want to read about weather in the boondocks. Who cares
about you, anyway? Sort of funny when you think of it that way, isn't it?

Bye guys,
Hildy Crowley










From: Walck. Scott D. :      walck-at-ppg.com
Date: Sat, 19 Feb 2000 16:42:48 -0500
Subject: DTSA Geometry setup for JEOL 2000FX in Experimental Header(Kevex

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Quantum)


Does anyone have the geometry settings for a Kevex Quantum detector on a
JEOL 2000FX? I am using DTSA and need the values. I would like to have the
sample to detector distance and the azimuthal angle. I am using +45 for the
azimuthal angle and 90 for the detector angle. Are these correct? I know
that the takeoff angle is 70.
-Scott


Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)






From: David Lockwood :      d.lockwood-at-mailbox.uq.edu.au
Date: Sun, 20 Feb 2000 09:13:45 +1000
Subject: Fluoro - GFP, Nikon filters, fixation

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Hi all. I have just joined this list and am posting without the usual
"lurking time" due to time constraints - please forgive if recently
covered. I am also a novice in all microscopy techniques, especially
fluorescence.

I am trying to detect GFP-containing cells in liver tissue and having
some problems.

1. I can't determine the spectra for the filter blocks I am using (on a
Nikon Eclipse TE300 with TE-FM Epifluorescence attachment). The blocks
are marked "UV", "B-2" (blue from source, green in field) and "G" (green
from source, red in field). I have e-mailed Nikon and to be fair
they've only had a few days but I'm under some pressure. No help from
the manual. I have been using B-2 for GFP.

2. I get quite a lot of background fluorescence even with frozen
sections (fixed in neutral buffered formalin). Can anyone suggest a way
to reduce this? (eg any extra filters?)

3. I have read conflicting opinions on fixation. Most previous work has
used thick sections (50 microns cut eg with Vibratome, ? to avoid having
to embed tissue blocks). GFP is interfered with by acetone (and probably
other organic solvents) but I would have thought that after fixation
with formalin it should be stabilized and resistant to the
xylene/alcohol used in paraffin embedding. I have seen fluorescence
retained after this treatment but perhaps it can be improved.

4. For those with liver fluoro experience - on "UV" setting I see bright
fluorescence which photobleaches. I believe I am looking at retinoids in
stellate cells, although the bleaching is incomplete and a bit slower
than I have seen before. Can anyone confirm this?

Apologies for long post. Any help much appreciated.

David Lockwood
University of Qld Dept of Surgery
PA Hospital Brisbane Australia




From: Marty Reed :      mmr7001-at-humboldt.edu
Date: Fri, 04 Jan 1980 08:23:24 -0800
Subject: questions for MSA members

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A colleague would like your responses to these questions. I believe that I
say something on the first question about a year ago and apologize for
asking again.


} Here's what I/we need to know.
}
} 1) Are Microstar diamond knives equal to Diatome. We have an offer that
} will save us about $1200 on three knives ( one 3mm ultra with 45 degree
} angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife).
}
}
} 2) Is the power controller feature on the Pelco Microwave Tissue processor
} (model #3451) worth it? (costs about $1300 additional compared to the
} Pelco 3450 without the power controller). I plan to do immunolocalization
} work, but am not sure this power controller feature is necessary or worth
} the cost.
}
} Casey
}
}
}
Thanks for your time
}
Marty Reed
Equipment Technician
Biology Department
Humboldt State University
Arcata CA 95521
707-826-3234
707-826-3201 FAX
mmr7001-at-axe.humboldt.edu









From: Dorota Wadowska :      wadowska-at-upei.ca
Date: Tue, 01 Feb 2000 10:58:08 -0400
Subject: TEM H600

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Hi!
Could anybody tell me what would be the sale price for a Hitachi
H600?
Thanks
Dorota




From: Walck. Scott D. :      walck-at-ppg.com
Date: Tue, 1 Feb 2000 11:28:02 -0500
Subject: RE: Question on Philips CM-12 SAD Alignment

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I don't use a CM series microscope on a regular basis, but what is important
is that you set it up the same way every time.

You can not make the back focal plane of the objective lens coincident with
the objective aperture. The back focal plane is well above the location of
the objective aperture plane.

There are two ways that I use to set up diffraction patterns reproducibly
depending on whether I am using CBED or SAD. Both are set up after the
sample has been made eucentric and the image focussed.

CBED: This method can be done for both CBED and SAD. The shadow of the
condenser aperture defines the diameter of the diffraction disk. When the
intermediate lens is adjusted properly, the edges of the diffraction disks
will be in focus. You are grabbing the back focal plane for your
diffraction pattern in the projector lens system. You will note that all of
the HOLZ lines (if you can see them) are the sharpest at this condition. If
you have a highly polycrystalline sample with continuous diffraction rings,
this method is difficult to do.

SAD. Spread the beam with the condenser all the way. (clockwise in the
CM-12 will go to more parallel beam faster than CCW -I think.) Then focus
the spot to the smallest that you can. You can take a really long exposure
or cheat a little and put some intensity back into the pattern with the
condenser lens. You will note that the objective aperture is not focused in
this method.

The most important thing to remember is to make the sample eucentric and
focus before during either of these methods and to do the diffraction
focussing consistently from sample to sample.
-Scott


Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)


"The opinions expressed are those of Scott D. Walck and not of PPG
Industries, Inc. nor of any PPG-associated companies."
--



} -----Original Message-----
} From: Valerie Leppert [mailto:vjleppert-at-ucdavis.edu]
} Sent: Monday, January 31, 2000 6:44 PM
} To: Microscopy-at-sparc5.microscopy.com
} Subject: Question on Philips CM-12 SAD Alignment
}
}
} --------------------------------------------------------------
} ----------
} 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 there;
}
} We have a new (to us) Philips CM-12 TEM in our lab and are
} wondering how
} to get the intermediate lens focused on the diffraction aperture (for
} making the first image plane and the diffraction aperture coincident
} prior to obtaining a SAED pattern). It doesn't seem to be
} covered in the
} manual.
}
} Any help would be appreciated as this is a completely new
} microscope to
} us.
}
} Thanks,
} Valerie Leppert
}
}




From: Laurie Wallin :      lwallin-at-ucsd.edu
Date: Tue, 1 Feb 2000 09:02:04 -0800
Subject: Any other fluorescence microscopy classes?

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I received Simon Watkins' note about the microscopy class in Maine and
wonder if anyone knows about similar courses closer to California in the
near future? We've got a number of technicians working on fluorescence
microscopy in our lab, and I think it would be great to have some more
formal training for us!

Sincerely,
Laurie Wallin
UCSD Department of Anesthesiology
9500 Gilman Drive, 0629, La Jolla, CA 92093




From: Robert S Dotson :      rdotson-at-mailhost.tcs.tulane.edu
Date: Tue, 1 Feb 2000 11:55:19 -0600 (CST)
Subject: used Philips EM410

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We are disposing of our old Philips 410 transmission
electron microscope. It is in pretty good shape but
is not fully functioning. At minimum it needs its ion
getter reconditioned. Does anyone know of someone who
might want to buy it and fix it up or use it for
parts?
-Robert


____________________________________

Robert S. Dotson, Ph.D.,
Laboratory Supervisor, Microscopy

Coordinated Instrumentation Facility
605 Lindy Boggs Building
Tulane University
6823 St. Charles Avenue
New Orleans, LA 70118-5698

504-865-5142| fax 504-865-6768

rdotson-at-mailhost.tcs.tulane.edu
http://www.tulane.edu/~cif
____________________________________





From: best-at-Juniata.Edu
Date: Tue, 1 Feb 2000 14:09:48 -0500
Subject: RE: weather & Formvar films

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I am thankful to those who took this weather thread and imparted
some information that I can really use. The retired professor that taught
me how to release forvar films had no idea why sometimes it worked and
sometimes it didn't. Now at least I have a couple of likely variables to
check.

Thanks!
Chris Best
Mol. Biol.
Juniata College
Huntingdon, PA 16652


PS - I can't help but be amused that even on a forum for scientists,
the petty &/or silly items get the most responses. Tell the truth, do you
stay up at night watching Jerry Springer? (For heaven sake, don't answer
that!)






From: Alwyn Eades :      jae5-at-lehigh.edu
Date: Tue, 01 Feb 2000 15:01:57 -0500
Subject: SAD and the back focal plane

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I am afraid that I do not agree with Scott Walck about back focal planes.
I do not have a CM 12 in the lab here. So I can not check that I am not
confusing the CM 12 with other Philips/FEI instruments. However, when
Scott says You can not make the back focal plane of the objective lens
coincident with the objective aperture. The back focal plane is well above
the location of the objective aperture plane. he is wrong. On all
microscopes except the very few which have very small pole-piece gaps for
high resolution, the objective aperture should coincide with the back focal
plane.

There is an easy and accurate way to find the true back focal plane on a CM
12 (or any other microscope with an immersion lens). Use a crystalline
sample, go to convergent-beam diffraction then use the diffraction focus to
make the Kikuchi lines as sharp as you can. That is the back focal plane.

If the microscope is set up properly, the image of the objective aperture
should be sharply in focus at nearly the same setting of the diffraction
focus. If the diffraction focus to give a sharp image of the objective
aperture is very different from the diffraction focus to make the Kikuchi
lines sharp, get your service engineer to reset the height of the objective
aperture until they agree.

If the sample is at the correct eucentric height, a selected-area
diffraction pattern in the true back focal plane will have sharp spots for
a C2 setting almost but not quite to the maximum (almost fully clockwise).
Again, if it does not, ask the service engineer to fix it.



Alwyn Eades
Department of Materials Science and Engineering
Lehigh University
5 East Packer Avenue
Bethlehem
Pennsylvannia 18015-3195
Phone 610 758 4231
Fax 610 758 4244
jae5-at-lehigh.edu





From: Walck. Scott D. :      walck-at-ppg.com
Date: Tue, 1 Feb 2000 15:58:05 -0500
Subject: RE: SAD and the back focal plane

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Alwyn,
We have a CM-12 at our other facility and I will check it out in a day or
so.

I thought that I did and it worked like our JEOL 2000FX. In the 2000FX,
because the lens is highly excited, there are 3 cross overs in the objective
lens after the sample. That means the true back focal plane is inside the
objective lens and it is not possible to put the aperture at that plane. In
the 2000FX, I have focused the CBED pattern and have found the objective
aperture not in focus. I have also found that the two methods that I
outlined do not agree with the camera constants. The 2000FX has a condenser
mini lens to make the beam parallel, but the highly excited lens allows the
small probes. Since the CM-12 can form the small probes, I thought that the
lens system was working in a similar manner.

I will try it out unless someone beats me to it. How about it CM owners?

-Scott


Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)



} -----Original Message-----
} From: Alwyn Eades [mailto:jae5-at-lehigh.edu]
} Sent: Tuesday, February 01, 2000 3:02 PM
} To: microscopy-at-sparc5.microscopy.com
} Subject: SAD and the back focal plane
}
}
} --------------------------------------------------------------
} ----------
} The Microscopy ListServer -- Sponsor: The Microscopy Society
} of America
} To Subscribe/Unsubscribe -- Send Email to
} ListServer-at-MSA.Microscopy.Com
} On-Line Help
} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
} --------------------------------------------------------------
} ---------.
}
}
}
} I am afraid that I do not agree with Scott Walck about back
} focal planes.
} I do not have a CM 12 in the lab here. So I can not check
} that I am not
} confusing the CM 12 with other Philips/FEI instruments.
} However, when
} Scott says "You can not make the back focal plane of the
} objective lens
} coincident with the objective aperture. The back focal plane
} is well above
} the location of the objective aperture plane." he is wrong. On all
} microscopes except the very few which have very small
} pole-piece gaps for
} high resolution, the objective aperture should coincide with
} the back focal
} plane.
}
} There is an easy and accurate way to find the true back focal
} plane on a CM
} 12 (or any other microscope with an immersion lens). Use a
} crystalline
} sample, go to convergent-beam diffraction then use the
} diffraction focus to
} make the Kikuchi lines as sharp as you can. That is the
} back focal plane.
}
} If the microscope is set up properly, the image of the
} objective aperture
} should be sharply in focus at nearly the same setting of the
} diffraction
} focus. If the diffraction focus to give a sharp image of
} the objective
} aperture is very different from the diffraction focus to make
} the Kikuchi
} lines sharp, get your service engineer to reset the height of
} the objective
} aperture until they agree.
}
} If the sample is at the correct eucentric height, a selected-area
} diffraction pattern in the true back focal plane will have
} sharp spots for
} a C2 setting almost but not quite to the maximum (almost
} fully clockwise).
} Again, if it does not, ask the service engineer to fix it.
}
}
}
} Alwyn Eades
} Department of Materials Science and Engineering
} Lehigh University
} 5 East Packer Avenue
} Bethlehem
} Pennsylvannia 18015-3195
} Phone 610 758 4231
} Fax 610 758 4244
} jae5-at-lehigh.edu
}
}




From: Henry Eichelberger :      heichelb-at-binghamton.edu
Date: Tue, 1 Feb 2000 16:14:07 -0500
Subject: RE: Reichert Ultracut parts & repair

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-----Original Message-----



Jordi:
For Reichert repair and parts I would suggest Helmut Patzig of MOC
(Microscopical Optical Consulting Inc.), phone (914) 268-6450 or e-mail
MOCLeica-at-Aol.com Helmut is known to carry spare parts for Reichert and LKB
ultramicrotomes.
I have no vested interest in MOC other than as a satisfied customer.
If he cannot help, you could ask him what your Reichert transformer voltage
output should be and using a voltage meter adjust the voltage of a variable
voltage transformer to the correct amount. Make sure to incorporate a
"stop" on the dial so the voltage can't be accidentally moved above the
correct amount.
Henry


Henry Eichelberger, Manager
Electron Microscopy Facility
Department of Biological Sciences
Binghamton University
Binghamton, NY 13902-6000

phone: (607) 777-2682
fax: (607) 777-6521
e-mail: heichelb-at-binghamton.edu


-----Original Message-----
------------------------------------------------------
On Mon, 24 Jan 2000, Marti, Jordi wrote:

} The power supply in our cryo microtome is having problems which might be
} related to the transformer. I was told by the service engineer that the
} transformer is no longer supported by Reichert. Does any one know where I
} can get a replacement ?
}
} Thanks
}
} Jordi Marti






From: Milo Kral :      m.kral-at-mech.canterbury.ac.nz
Date: Wed, 02 Feb 2000 15:28:32 +1300
Subject: Looking for a turbo pump

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Does anyone have an extra turbo pump that they can part with?

the weather is quite wintery considering its summer here in Christchurch
New Zealand. low clouds with southerly prevailing winds (explaining the
COLD)






From: Garber, Charles A. :      cgarber-at-2spi.com
Date: Wed, 02 Feb 2000 02:37:10 -0500
Subject: Silver membranes being discontinued

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-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --

Hello,

I apologize in advance if this posting offends anyone. However there are a
good many people who use silver membranes in their work and to have the
supply from the worlds only manufacturer (to my knowledge) come to a halt,
has the potential of being highly disruptive at least to some programs:
=================================================
Osmonics has announced the closing of our Phoenix, AZ manufacturing facility
effective 1 May 2000. Since our silver membranes are manufactured in this
facility, Osmonics has decided to end production of this membrane because of
declining sales and the very expensive costs associated with moving the mfg.
. plant to another location. All orders placed before 1 March, 2000 will be
honored and filled.
==================================================
We plan to make a "last buy" before the cut off date of March 1, 2000. I
would advise anyone depending on these silver membranes for their work to
take stock of their future requirements because after the cut off date,
sales will be possible only from remaining stocks.

For those who do run out, and are in a bind, we are prepared help them find
alternative filtration media, of which there are indeed some alternatives
for some applications.

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.spi.cc
########################
============================================




From: Keith Ryan :      KPR-at-wpo.nerc.ac.uk
Date: Wed, 02 Feb 2000 08:35:01 +0000
Subject: LM - Cryostat - marine larvae

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Hello Listers

I have someone here who wants to freeze and cut cryostat sections of
marine larvae (5 microns?) for immuno/light microscopy. The specimens
are 100-200 microns in length.

1. What would be the best way of handling these small items?
2. What would be the best support/medium e.g TissueTek?
3. What about cryoprotection? I am familiar with 2.3 molar sucrose
for EM.
4. Any general tips for immuno (protein/amino peptidases)?

Thanks - Keith
_______________________
Keith Ryan (Dr)
Marine Biological Association
Citadel Hill
Plymouth
Devon PL1 2PB
England

Tel. 0044 (0)1752 633279 (with answer machine)
also 0044 (0)1752 633249
Fax. 0044 (0)1752 633102

e-mail: kpr-at-wpo.nerc.ac.uk





From: Ron Doole :      ron.doole-at-materials.oxford.ac.uk
Date: Wed, 2 Feb 2000 08:38:24 +0000 (GMT Standard Time)
Subject: re - CM12 SAD focus

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Hi Valerie,

Like the other respondees I am not a CM12 user
(where are they?), however, as an ex CM12 user of some 10
years ago I think that I remember the alignment that Valerie
is asking about. In the basic alignment of the machine there
was a step during which the SAD aperture was focussed. I
think that it was in the `service calibrations' page. This
may have changed in later software revisions.

The alignment of this microscope was usually
carried out by the engineer when the instrument was
installed. They would set up the objective lens current and
adjust the specimen goniometer height to ensure that when
it was at the eucentric position it was correctly in focus.
Following this a complete column alignment was carried out
including focussing the SAD aperture. The manufacturers
then assumed that the operator would set up the specimen to
the eucentric height, it should focus in the same position
and the aperture should be in focus.

If the aperture is not in focus when in the SA
range (as noted next to the mag readout) then check you are
correctly at the eucentric position, if you are then either
it was not aligned properly after installation or something
has changed.
NOTE: DO NOT ADJUST THE SERVICE ALIGNMENTS UNLESS YOU KNOW
WHAT YOU ARE DOING AS YOU CAN LOSE THE INSTRUMENT
ALIGNMENTS.

Good luck,
Ron


----------------------
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





From: Jonathan Barnard :      Jonathan.Barnard-at-Angstrom.UU.SE
Date: Wed, 02 Feb 2000 11:20:15 +0100
Subject: TEM:finding the parallel beam

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I got a tip for finding the condenser lens settings required for parallel
illumination from Angus Kirkland (Cambridge).

Make sure you are in microprobe mode and set up for SAED and the specimen
is out of the way. Spread the beam, put in the SA Aperture, go to
diffraction and then sweep the SA aperture (SAA) from side to side and
minimise the deflection of the diffraction spot by tweaking the
illumination control (C2 or second condenser lens). A little bit of thought
will convince you that anything other than a parallel beam will give you a
side to side motion (tilt) when the SAA is swept.

I have found it useful to keep a table of C2 condenser lens current
settings for each spot size (C1 excitation) in microprobe mode. Setting the
C2 lens for parallel illumination and adjusting the diffraction focus to
get the sharpest spot will then give you near perfect SAD conditions.

Anyone contemplating electron holography for example, will have to start
from the parallel illumination to get the best spatial coherence for their
holograms.

********************************************************
Dr Jonathan Barnard

Analytical Materials Physics
The Angstrom Laboratory, Uppsala University
P O Box 534, SE-751 21 Uppsala, Sweden
Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
http://www.angstrom.uu.se/analytical/home.html

********************************************************





From: Matt_Plantinga-at-amway.com
Date: Wed, 2 Feb 2000 08:15:48 -0500
Subject: LSCM Need help with non-fluorescing resin

Contents Retrieved from Microscopy Listserver Archives
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I am working on fluorescent imaging of thin sections cut from samples
embedded in TEM embedding resin, and am having a significant problem with
resin fluorescence. Does anyone know of an embedding resin that doesn't
autofluoresce?

Matt Plantinga
Amway Corporation
matt_plantinga-at-amway.com





From: Andrew Ochalski :      aochalsk-at-science.uottawa.ca
Date: Wed, 2 Feb 2000 14:05:21 GMT+5
Subject: Heating stage with DvorakStotler Chamber...Recommendations?

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{color} {param} 0100,0100,0100 {/param} {FontFamily} {param} Arial {/param} {smaller} Fellow Microscopists,


We have a Zeiss Axiophot microscope and a
Dvorak-Stotler Chamber that we would like to
assemble into a temperature-controlled environment
for digital live-cell imaging. I have several questions
about the best way to combine these elements:

i) Can anyone recommend a heating stage
compatible with both the microscope and the
chamber?

ii) Would the best position for a temperature sensor
be on the top (near the objective?) or on the bottom
of the chamber, or both (two sensors)?

iii) Is it better to heat the media/fluids in their
containers, or to have an in-line heater ( brand
recommendations?)

iv) Are there any fuid flow or temperature
considerations specific to this chamber that need to
be addressed (gravity vs. pump feed, shear forces,
heating stage redundant, etc.)?

v) Are there any problems related to objective lens
mag/NA that need to be overcome when using this D-
S apparatus?


Any references, anecdotes, or words of wisdom
would be appreciated. Dealers, if you have a product
that you think I could use, please don't hesitate to
contact me. Thank you in advance. {FontFamily} {param} BrushScript BT {/param} {bigger}

{nofill}
Regards,

Andrew Ochalski,
Microscopy Technician,
Dept. of Biology,
University of Ottawa
Room 108, Gendron Bldg.
130 Louis Pasteur,
Ottawa, Ontario
CANADA
K1N 6N5
613-562-5800 x6343
FAX: 613-562 5486

{/x-rich}



From: Cheri Owen :      cowen-at-cmb.biosci.wayne.edu
Date: Wed, 02 Feb 2000 09:32:01 -0500
Subject: Looking for a new microscope

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I would like to get some information from some of the top companies on
microscopes with photographic and possibly fluourescent capabilities.
Please email me or call 313-993-4195
Thank you

Cheri Owen
Wayne State University





From: BARRY SHAW :      Barry.Shaw-at-nottingham.ac.uk
Date: Wed, 2 Feb 2000 14:36:36 GMT0BST
Subject: Re: Any other fluorescence microscopy classes?

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Hello All,

Not really a reply but another question!!
Anybody know of a similar course a little further east - England
(Great Britain) to be exact !!?
Thanks in Advance,

Baz


Barry Shaw
Senior Imaging Technician
Molecular & Cell Biology
E Floor
School Of Biomedical Sciences
University of Nottingham Medical School
NG7 2UH




From: Manfred Prantl :      prantl-at-alicona.com
Date: Wed, 02 Feb 2000 15:42:52 +0100
Subject: SEM and 3D?

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We are a company that has specialized on extracting 3D information
from stereoscopic SEM images. Currently we try to figure out the
potential applications and the future prospects of such techniques for
the microscopy community.

What we do exactly is that we take two images from a sample on the
SEM from slightly different tilt angles and use digital image processing

to compute a 3D elevation model. From there you can then perform
various analysis steps like extraction of 3D profiles, determination
of surface roughness etc.

My questions now are:

1.) How would you judge the importance of obtaining 3D data from a SEM
image?
2.) What would be the major requirements for such a data set to be
useful
(like density, accuracy, number of false alarms, etc.)?
3.) Is it of importance to you that you get the 3D data and the image
data
together and not separated (like it is the case with AFM)?
4.) How would you judge the future prospects and influence of such a
technique on your personal field of work?
5.) Do you generally trust the measurement of 3D data via stereoscopic
images?
6.) Do you already have experience with stereoscopic depth measurments
and what are your conclusions?

Best regards, Manfred Prantl
R&D
Alicona GmbH, Germany
www.alicona.com





From: Russell E. Cook :      cook-at-horus.et.anl.gov
Date: Wed, 2 Feb 2000 09:15:18 -0600
Subject: Back focal plane of CM's

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Irene Piscopo of FEI/Philips probably can answer your question. I'm copying
this message to her although if she's in the "field" a response may take a
few days. Also, Max Otten of FEI/Philips is another good source for that
type of information.

We have a CM-120 so we're aware of your problems in deciphering the
operator's manual. Good luck!

Bruce F. Ingber, Biologist
USDA-ARS, SRRC
1100 Robert E. Lee Blvd.
New Orleans, LA 70124

(504) 286-4270 phone
(504) 286-4419 fax
bingber-at-nola.srrc.usda.gov

-----Original Message-----
} From: Valerie Leppert [mailto:vjleppert-at-ucdavis.edu]
Sent: Monday, January 31, 2000 5:44 PM
To: Microscopy-at-sparc5.Microscopy.Com


I can't speak about Philips CM12's, but the back focal plane of the
objective coincides with the objective aperture in the CM30 here.

----------------------------------------------------------------------
Russell E. Cook, Ph.D.
Electron Microscopy Center for Materials Research
Argonne National Laboratory
Materials Science Division, Building 212
9700 South Cass Avenue
Argonne, IL 60439-4838
(630)252-7194
FAX: (630)252-4289
cook-at-horus.msd.anl.gov






From: Piscopo, Irene :      IPiscopo-at-FEICO.COM
Date: Wed, 2 Feb 2000 07:59:45 -0800
Subject: RE: Question on Philips CM-12 SAD Alignment

Contents Retrieved from Microscopy Listserver Archives
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Hello all: Valerie Leppart, you can reach me at 203-853-3256. I cannot call
you since there is no phone number to contact. Irene Piscopo

The CM 12 microscope has four mag ranges: LM, M, SA, Mh

As long as you are eucentric and using the SA range the image plane
and the diffraction aperture plane will be in focus. (It is done
automatically by the instrument.) This will give you accurate SAD down to
1um. The objective mag in the plane of the diffraction aperture is
approxmately 27X. If you wish to obtain diffraction from areas smaller than
one micron, use uD, or uuD. If you call me I will discuss these methods
with you and send you detailed instructions on using these methods.


Irene Piscopo


} -----Original Message-----
} From: Ingber, Bruce F. [SMTP:bingber-at-commserver.srrc.usda.gov]
} Sent: Wednesday, February 02, 2000 10:14 AM
} To: Microscopy-at-MSA. Microscopy. com (E-mail)
} Cc: Piscopo Irene (CS) (E-mail); Max T. Otten (E-mail)
} Subject: FW: Question on Philips CM-12 SAD Alignment
}
} Irene Piscopo of FEI/Philips probably can answer your question. I'm
} copying
} this message to her although if she's in the "field" a response may take a
} few days. Also, Max Otten of FEI/Philips is another good source for that
} type of information.
}
} We have a CM-120 so we're aware of your problems in deciphering the
} operator's manual. Good luck!
}
} Bruce F. Ingber, Biologist
} USDA-ARS, SRRC
} 1100 Robert E. Lee Blvd.
} New Orleans, LA 70124
}
} (504) 286-4270 phone
} (504) 286-4419 fax
} bingber-at-nola.srrc.usda.gov
}
} -----Original Message-----
} From: Valerie Leppert [mailto:vjleppert-at-ucdavis.edu]
} Sent: Monday, January 31, 2000 5:44 PM
} To: Microscopy-at-sparc5.Microscopy.Com
} Subject: Question on Philips CM-12 SAD Alignment
}
} ------------------------------------------------------------------------
} 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 there;
} We have a new (to us) Philips CM-12 TEM in our lab and are wondering how
} to
} get the intermediate lens focused on the diffraction aperture (for making
} the first image plane and the diffraction aperture coincident prior to
} obtaining a SAED pattern). It doesn't seem to be covered in the manual.
} Any help would be appreciated as this is a completely new microscope to
} us.
} Thanks,
} Valerie Leppert




From: rgriffin-at-eng.uab.edu
Date: Wed, 2 Feb 2000 14:15:55 -0600
Subject: Gray level

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How many gray levels can the human eye distinguish? We've found several
references that disagree.
If anyone knows of a reference - that would be best.


Robin Griffin
UAB




From: Hendrik O. Colijn :      colijn.1-at-osu.edu
Date: Wed, 02 Feb 2000 16:07:50 -0500
Subject: Re: Gray level

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Robin,

I don't have the reference right in front of me, but I believe that
Gonzalez and Wintz say that

1. The eye can respond to an intensity range of ~10^10 in light intensity,
but this is the adaptive response. The perceived brightness is a log (some
will argue power law) function of the incident intensity.

2. In any one point in an image, the eye can distinguish at most ~20-30
gray levels... But... in a complex image you need at least 100 gray levels
for the eye to see it as smooth. In other words, the eye seems to adapt as
it scans the image.

Cheers,
Henk


At 02:15 PM 2/2/00 -0600, rgriffin-at-eng.uab.edu"-at-sparc5.Microscopy.Com wrote:

} How many gray levels can the human eye distinguish? We've found several
} references that disagree.
} If anyone knows of a reference - that would be best.
}
}
} Robin Griffin
} UAB

Hendrik O. Colijn colijn.1-at-osu.edu
Campus Electron Optics Facility Ohio State University
(614) 292-0674 http://web.ceof.ohio-state.edu
Murphy's Law: "If anything can go wrong, it will."
Commentary: "Murphy was an optimist."





From: Valerie Leppert :      vjleppert-at-ucdavis.edu
Date: Wed, 2 Feb 2000 15:04:41 -0800 (PST)
Subject: Re: re - CM12 SAD focus

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Thanks for your reply and everyone else's reply regarding the SAD
alignment. There seem to be a lot of different opinions about how this
done!

We did set the sample at the eucentric and did notice that the SAD
aperture is not quite in focus when in image mode. Of course this will
cause the diffraction information to come from an area that is displaced
with respect to the aperture position.

I am accustomed to being able to independently focus the intermediate lens
on the SAD aperture, and then bring the image into focus using the
objective lens - making the image plane and the SAD aperture coincident.
This is on a much older Philips model and a much older Hitachi model.

However, on the CM-12, there doesn't seem to be a way for the user to do
this in normal operation. I've tried a few of the suggestions (haven't
worked my way through them all yet!) with no luck yet.

It does seem that this adjustment has been grouped under the category of
"service alignment" in newer models.

Thank you to everyone who has replied.

Valerie Leppert






From: Karl Hagglund-KW :      hagglund.kw-at-pg.com (by way of Nestor J. Zaluzec)
Date: Wed, 2 Feb 2000 17:06:39 -0600
Subject: LM Sony camera bayonet mount

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We have a sony DXC-760 MD ccd camera that we have mounted to a standard camera
lens with a bayonet adapter. Unfortunately, we have borrowed the adapter and
have to return it. Is there a standard size (i.e. Nikon, Minolta, Canon) lens
adapter that works with this camera mount? We can't find a part number or
description for the adapter, and the book doesn't describe the size. We have
already ordered and returned one that didn't fit.

Thanks in advance.

Karl Hagglund
Proctor and Gamble
Food and Beverage Analytical Microbiology






From: Dr Eric Lachowski :      che136-at-abdn.ac.uk (by way of Nestor J. Zaluzec)
Date: Wed, 2 Feb 2000 17:14:03 -0600
Subject: Electron diffraction simulations

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Fellow Listers,
Does anyone know of a (preferrably free) software package
which simulates ray paths, including the diffraction mode,
in the TEM? We would like to use such a package to teach
physics students a bit of optics.

Thanks,
Eric



----------------------
Dr Eric Lachowski
Department of Chemistry
University of Aberdeen
Meston Walk
Old Aberdeen AB24 3UE
Scotland
+44 (0)1224 272934 fax 272921
e.lachowski-at-abdn.ac.uk






From: Paul Webster :      pwebster-at-mailhouse.hei.org
Date: 02 Feb 00 16:29:17 -0800
Subject: RE: LSCM Need help with non-fluorescing resin

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The Microscopy ListServer -- Sponsor: The Microscopy Society of America


Reply to: RE: LSCM Need help with non-fluorescing resin
Dear Matt,

Try any of the immunocytochemical resins (Lowicryl, LR White or Gold, Unicryl etc). When specific antibodies are used on embedded tissue, they produce very beautiful fluorescent patterns that look like line drawings.

Paul Webster

Matt_Plantinga-at-amway.com wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
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} -----------------------------------------------------------------------.
}
}
} I am working on fluorescent imaging of thin sections cut from samples
} embedded in TEM embedding resin, and am having a significant problem with
} resin fluorescence. Does anyone know of an embedding resin that doesn't
} autofluoresce?
}
} Matt Plantinga
} Amway Corporation
} matt_plantinga-at-amway.com
}
}
}
}
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fax: 213 413 6739
e-mail: pwebster-at-hei.org
http://www.hei.org/htm/aemi.htm





From: DrJohnRuss-at-aol.com
Date: Wed, 2 Feb 2000 19:56:04 EST
Subject: Re: Gray level

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In a message dated 2/2/00 4:00:26 PM,
rgriffin-at-eng.uab.edu-at-sparc5.Microscopy.Com writes:

} How many gray levels can the human eye distinguish? We've found several
}
} references that disagree.
}
} If anyone knows of a reference - that would be best.

Most of the books on image processing say the same thing (probably all copied
from each other and other non-prime sources), but I don't know of an original
reference based on real research. You can find the same ideas in books like
Frisby "Seeing: illusion, brain and mind", Oxford, 1980; Marr "Vision",
Freeman, 1982; and Rock "Perception", Freeman, 1984 which deal with the
human visual system not in the context of computer graphics. These are
probably closer to the original research.

The basic idea is that it takes about a 2-2.5% change in absolute brightness
(i.e. a ratio of 1.025) to be just detectable, and only if it is a fairly
sudden change spatially (or temporally for that matter). That is where the
20-30 distinct brightness levels over the full range of brightness that can
be seen at one time (i.e. without accommodations like varying pupil size)
comes from. The requirement that an entire scene have about 100 levels to
avoid banding is more controversial, based on the idea that the eye can
perform some accommodation as it traverses from one part of a scene to
another. This probably doesn't apply when (e.g.) looking through a
microscope, so the 20-30 number would be more applicable. It also ignores
color, which complicates things further.




From: special123-at-smartportfolio.com
Date: Thu, 3 Feb 2000 12:17:58 +0900
Subject: TEM:finding the parallel beam

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Dear Jonathan,

} From what is my understanding of optics - this will not give you parallel
illumination at the specimen plane (if this what you meant?).
By tweaking C2 you are moving the crossover plane (which is actually the
diffraction plane). You can in the same way make the spot not to move by
changing the diffraction focus. For each setting of C2 you can find the
crossover by changing the diff. focus.
Try this - spread the beam, go to diffraction, focus by diffraction focus
for smallest spot, go back to imaging mode put the SAA, go to diffraction
and try the procedure you described. I think the spot will not move.
Ofcourse if you expand the beam too much you can no longer produce small
diffraction spot (without the use of SAA) because you go out of paraxial
mode.
The diffraction plane is always the crossover plane not the theoretical back
focal plane of the objective. Its z-position changes with the change of the
C2 excitement.
The spatial coherency of the electron waves is very high (actually the
electrons are flying one by one through the microscope). The limiting
factors are the source size, the illumination angle and the energy spread.

Please correct me if I'm wrong ... I'm still "green" in electron microscopy
.. I'll be happy to learn from experienced users.

Best wishes,

Rado

---------------------------------------------------------------------
Radostin Danev
Laboratory of Ultrastructure Research
National Institute for Physiological Sciences
Myodaiji-cho, Okazaki 444-8585, JAPAN
e-mail: rado-at-nips.ac.jp
---------------------------------------------------------------------
----- Original Message -----
} From: Jonathan Barnard {Jonathan.Barnard-at-Angstrom.UU.SE}
To: {microscopy-at-sparc5.microscopy.com}
Sent: Wednesday, February 02, 2000 7:20 PM


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From: Dmitri V. Sokolov :      sokolov-at-ryouko.rciqe.hokudai.ac.jp
Date: Thu, 3 Feb 2000 20:20:06 +0900
Subject: JAMP-7810 manual in English and local chemical analysis

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Dear all,

We are making preparations for experiments on determination of the chemical
composition of semiconductor oxides, formed locally. The size of the
modified area should be about 1 micron, and we are not sure yet if it is
possible to make mentioned chemical analysis with Auger microprobe at all.

Another problem is that our newly came Auger spectroscope JAMP-7810 has no
English manual with it. And believe me, it is barely possible to read that
in Japanese!

We would be very grateful for your suggestions on both my questions:
- how to measure the best chemical bonding on the spot with 1x1 sq.
micrometer dimensions
- where to get the manual (or copy of it) from.
Remark: Mails to USA and Japanese representatives of JEOL gave no result!

Best regards.
Dmitri


__________________________________________
Dmitri V. Sokolov, Doctor Course student
Research Center for Interface Quantum Electronics,
Hokkaido University, North 13, West 8, Kitaku,
Sapporo 060, Hokkaido, Japan
Phone 81-11-706-7174 Fax 81-11-716-6004
http://www.geocities.com/SiliconValley/Campus/1314
AOL Instant Messenger FalconDot
ICQ 9418072
__________________________________________






From: bengi :      bengi-at-unimo.it
Date: Thu, 3 Feb 2000 14:54:20 +0100
Subject: Nitrogen determination

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Dear listers,
I have started a research project on ammonium -bearing mineral (zeolites),
and now I'm trying to quantify N using electron microprobe analysis. My
problems of dealing with N are:

a) The thickness of the coating (in the literateur = approximately 150 A).
Does anyone have information on the model of coater to making these films?
Does anyone have experience with preparation of such
samples?

b) Obtaining appropriate standards. (I have Si3N4, but it contains too much
N (=39.96%) towards my samples (5%). Does anyone known any good nitrogen
standards for such a sample ?

Any other suggestions about quantitative nitrogen analysis would be greatly
appreciated !!
Thanks for your time.

My best regards

Eugenia


Eugenia Marchi
Universit degli Studi di Modena
Dip. Scienze della Terra
P.le S.Eufemia n.19
41100 Modena (Italy)
Tel. +39-059-417289
Fax. +39-059-417399
e-mail: bengi-at-unimo.it






From: Barr, Dennis B :      dennbarr-at-eastman.com
Date: Thu, 3 Feb 2000 09:30:07 -0500
Subject: RE: Silver membranes being discontinued

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It's unfortunate that this manufacturer is stopping production. Aren't
there other manufacturers?

Dennis B. Barr (dennbarr-at-eastman.com)
Physical Chemistry Research Laboratory
Physical & Analytical Chemistry Research Division
Eastman Chemical Company
Kingsport, TN 37662-5150

B-150B, R-132E, (423) 229-2188


} -----Original Message-----
} From: Garber, Charles A. [SMTP:cgarber-at-2spi.com]
} Sent: Wednesday, February 02, 2000 2:37 AM
} To: MICROSCOPY BB
} Subject: Silver membranes being discontinued
}
} ------------------------------------------------------------------------
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}
}
} -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
}
} Hello,
}
} I apologize in advance if this posting offends anyone. However there are a
} good many people who use silver membranes in their work and to have the
} supply from the worlds only manufacturer (to my knowledge) come to a halt,
} has the potential of being highly disruptive at least to some programs:
} =================================================
} Osmonics has announced the closing of our Phoenix, AZ manufacturing
} facility
} effective 1 May 2000. Since our silver membranes are manufactured in this
} facility, Osmonics has decided to end production of this membrane because
} of
} declining sales and the very expensive costs associated with moving the
} mfg.
} .. plant to another location. All orders placed before 1 March, 2000 will
} be
} honored and filled.
} ==================================================
} We plan to make a "last buy" before the cut off date of March 1, 2000. I
} would advise anyone depending on these silver membranes for their work to
} take stock of their future requirements because after the cut off date,
} sales will be possible only from remaining stocks.
}
} For those who do run out, and are in a bind, we are prepared help them
} find
} alternative filtration media, of which there are indeed some alternatives
} for some applications.
}
} 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.spi.cc
} ########################
} ============================================




From: Michael Bode :      mb-at-soft-imaging.com
Date: Thu, 3 Feb 2000 08:11:02 -0700
Subject: RE: Gray level

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Yes, I agree with John. A better question than "how many gray levels can
one see" is probably "how many gray levels can one see under what
circumstances". The answer is probably very different for a static image
and a dynamic image, with edges or without, in low light conditions or
bright light conditions.

The human eye (and brain) has had Millions of years to evolve and
develop some rather sophisticated algorithms to deal with different
conditions and the answer is probably also very complex. For example,
our vision is extremely good at detecting movement. On a completely
static image this tends to depress differences, while on a dynamic image
it tends to enhance differences (we automatically focus on the changes).


Maybe you can test that yourself:

take the same image on a computer at different bit depths (=levels of
gray). Then randomly put them on the screen (without watching the
change) and try to decide, which image it is. Then do the same but watch
the changes and try to decide how many gray levels you need before the
changeover becomes invisible. You could do this with a "noise" image, an
image that shows some object, and a smooth gray wedge. I for one would
be interested to hear about the results.

Michael

Michael Bode, Ph.D.
Soft Imaging System Corp.
1675 Carr St., #105N
Lakewood, CO 80215
===================================
phone: (888) FIND SIS
(303) 234-9270
fax: (303) 234-9271
email: mailto:info-at-soft-imaging.com
web: http://www.soft-imaging.com
===================================



} ----------
} From:
"DrJohnRuss-at-aol.com"-at-sparc5.Microscopy.Com[SMTP:"DRJOHNRUSS-at-AOL.COM"-at-SPA
RC5.MICROSCOPY.COM]
} Sent: Wednesday, February 02, 2000 5:56:04 PM
} To: Microscopy-at-sparc5.Microscopy.Com; rgriffin-at-eng.uab.edu
} Subject: Re: Gray level
} Auto forwarded by a Rule
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In a message dated 2/2/00 4:00:26 PM,
rgriffin-at-eng.uab.edu-at-sparc5.Microscopy.Com writes:

} How many gray levels can the human eye distinguish? We've found
several
}
} references that disagree.
}
} If anyone knows of a reference - that would be best.

Most of the books on image processing say the same thing (probably all
copied
from each other and other non-prime sources), but I don't know of an
original
reference based on real research. You can find the same ideas in books
like
Frisby "Seeing: illusion, brain and mind", Oxford, 1980; Marr "Vision",

Freeman, 1982; and Rock "Perception", Freeman, 1984 which deal with the

human visual system not in the context of computer graphics. These are
probably closer to the original research.

The basic idea is that it takes about a 2-2.5% change in absolute
brightness
(i.e. a ratio of 1.025) to be just detectable, and only if it is a
fairly
sudden change spatially (or temporally for that matter). That is where
the
20-30 distinct brightness levels over the full range of brightness that
can
be seen at one time (i.e. without accommodations like varying pupil
size)
comes from. The requirement that an entire scene have about 100 levels
to
avoid banding is more controversial, based on the idea that the eye can
perform some accommodation as it traverses from one part of a scene to
another. This probably doesn't apply when (e.g.) looking through a
microscope, so the 20-30 number would be more applicable. It also
ignores
color, which complicates things further.




From: MICHAEL MOHN :      MMOHN-at-mail.monroe.cc.mi.us
Date: Thu, 03 Feb 2000 10:14:13 -0500
Subject: Re: LM Sony camera bayonet mount

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Most industrial cameras use C-mounts. C-mount to camera lens adapters are available from Edmund Industrial Optics (http://www.edmundoptics.com phone:800-363-1992) on page 165 of their 2000 catalog. They have mounts for several types of cameras, and list for $62 each. If your connecting the camera to a 'scope, Edmund sells a relay lens for $240 which mounts the camera to a 23mm eyepiece tube. For connecting to trinocular and CCD ports on a scope, Diagnostic Instruments sells relay lenses for many types of 'scopes (http://www.diaginc.com) for $299.


Michael Mohn
Assistant Professor of Materials Technology
Monroe County Community College
Phone: 734-384-4122 Fax: 734-242-9711
http://www.monroe.cc.mi.us/mmohn

} } } "Karl Hagglund-KW" by way of Nestor J. Zaluzec {hagglund.kw-at-pg.com} 02/02 6:06 PM } } }
We have a sony DXC-760 MD ccd camera that we have mounted to a standard camera
lens with a bayonet adapter. Unfortunately, we have borrowed the adapter and
have to return it. Is there a standard size (i.e. Nikon, Minolta, Canon) lens
adapter that works with this camera mount? We can't find a part number or
description for the adapter, and the book doesn't describe the size. We have
already ordered and returned one that didn't fit.

Thanks in advance.

Karl Hagglund
Proctor and Gamble
Food and Beverage Analytical Microbiology








From: rlvaughn-at-unmc.edu
Date: Thu, 3 Feb 2000 09:39:47 -0600
Subject: Re: Gray level

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Are you equating brightness levels with grey levels here?
I was told something like this years ago when we were buying our first
digital B&W camera, when I asked the vendor about the competitors thousand
plus grey levels vs his 256 grey levels and was told that the eye could not
distinguish greater levels. Now, four or five years later everyone is
pitching 10, 12 and 16? bit cameras, and yet the images from these cameras
do look better. Is it the bits and increased grey levels? I know that if
your doing image analysis at the pixel level that more bits are to your
advantage but what about just photgraphic quality?

Rick Vaughn





From: L. D. Marks :      ldm-at-apollo.numis.nwu.edu
Date: Thu, 3 Feb 2000 10:06:36 -0600 (CST )
Subject: Re: TEM:finding the parallel beam

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Sorry, but I think we are getting a little confused here.

1) The diffraction plane is (by definition) the objective lens
focal length below the sample - it is the focus of parallel
directions coming from the sample. The focal length (and
parameters such as Cs, Cc) change rapidly with the objective lens
current so an eucentric position makes life simple but is not
necessarily the best condition to use. (For HREM you generaly
want to have the objective lens stronger and reduce Cs.)

2) In general C2 does not change the focal length at all. You
can have some effect from coupling of the magnetic fields, but
all C2 is supposed to do is change the convergence angle.

3) The objective aperature is (approximately) in the back-focal
plane, given that it is finite and height adjustments are only
done coarsely (for a specific value of the objective current
only).

4) The selected area aperature is (approximately) in the same
plane as the object for the same reasons as above. Remember that
there are positional errors in SA diffraction due to Cs (see
Hirsch et al for instance).

5) You should focus Kikuchi-lines, not go for the smallest spot,
since these are real object in the diffraction pattern. What you
will see is then representative of you incident beam, sometimes
a Gaussian range of directions.

6) The simplest way to get more parallel illumination in general is
to reduce the condensor aperture size.

7) All bets are off if you have a FEG. Rather than having incoherent
illumination things like the Cs of the prefield (above the sample)
and postfield (below) add coherently making it much more complicated.
For instance, the illumination angle varies across the field of view.
While this is also there with LaB6, it is a weak effect (except for
certain classes of diffracted beams, e.g. those with a screw-axis). With
a FEG you can get a "spot" pattern many ways, and most of these will have
large aberrations (e.g. pincushion).

++++++++++++++++++++++++++++++++++++++++++++++++
Laurence Marks
Department of Materials Science and Engineering
Northwestern University
fax: (847) 491-7820
mailto:l-marks-at-nwu.edu
http://www.numis.nwu.edu
++++++++++++++++++++++++++++++++++++++++++++++++





From: Dr Eric Lachowski :      che136-at-abdn.ac.uk
Date: Thu, 3 Feb 2000 16:24:08 +0000 (GMT)
Subject: Re: Nitrogen determination

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Eugenia,
the microprobe is not the best tool for the analysis of
ammonia in zeolites because ammonia is very volatile in the
electron beam. If your zeolite is phase pure it would be
far better to dissolve it and do a classical chemical
analysis.

regards,
Eric
On Thu, 3 Feb 2000 14:54:20 +0100 bengi {bengi-at-unimo.it}
wrote:


} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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} -----------------------------------------------------------------------.
}
}
}
}
} Dear listers,
} I have started a research project on ammonium -bearing mineral (zeolites),
} and now I'm trying to quantify N using electron microprobe analysis. My
} problems of dealing with N are:
}
} a) The thickness of the coating (in the literateur = approximately 150 A).
} Does anyone have information on the model of coater to making these films?
} Does anyone have experience with preparation of such
} samples?
}
} b) Obtaining appropriate standards. (I have Si3N4, but it contains too much
} N (=39.96%) towards my samples (5%). Does anyone known any good nitrogen
} standards for such a sample ?
}
} Any other suggestions about quantitative nitrogen analysis would be greatly
} appreciated !!
} Thanks for your time.
}
} My best regards
}
} Eugenia
}
}
} Eugenia Marchi
} Universit degli Studi di Modena
} Dip. Scienze della Terra
} P.le S.Eufemia n.19
} 41100 Modena (Italy)
} Tel. +39-059-417289
} Fax. +39-059-417399
} e-mail: bengi-at-unimo.it
}
}
}

----------------------
Dr Eric Lachowski
Department of Chemistry
University of Aberdeen
Meston Walk
Old Aberdeen AB24 3UE
Scotland
+44 (0)1224 272934 fax 272921
e.lachowski-at-abdn.ac.uk







From: L. Paul Bdard :      Paul_Bedard-at-uqac.uquebec.ca
Date: Thu, 3 Feb 2000 12:12:17 -0500
Subject: Need repairman for Hitachi SEM

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Hello,

I need somebody to repair a Hitachi SEM S-2700.
Please contact me individually not the list.
Thanks for help,
--
L.Paul Bdard, ing. Ph.D.
Resp. Lab. Gochimie | Lab. Manager
Sciences Appliques ; Universit du Qubec Chicoutimi Canada
Tl. : 418/545-5011 x 2276 | Alt. E-mail : pbedard-at-saglac.qc.ca
{} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}
La priode humaine, depuis la cration d'Adam jusqu' nos jours, n'est qu'une
moisissure dans l'histoire de notre plante
JCK Laflamme 1886




From: Warren E Straszheim :      wesaia-at-iastate.edu
Date: Thu, 03 Feb 2000 11:22:46 -0600
Subject: Re: Gray level

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Let me offer my guess that the improved appearance of the images is be due
largely to the improved signal-to-noise ratio in the newer cameras.

Of course there is also something to be gained in the dynamic range by the
extra bits of resolution. They are much more forgiving and allow
substantial changes in contrast and brightness without sacrificing the
information contained in the data.

Warren S.

At 09:39 AM 2/3/2000 -0600, you wrote:

} Are you equating brightness levels with grey levels here?
} I was told something like this years ago when we were buying our first
} digital B&W camera, when I asked the vendor about the competitors thousand
} plus grey levels vs his 256 grey levels and was told that the eye could not
} distinguish greater levels. Now, four or five years later everyone is
} pitching 10, 12 and 16? bit cameras, and yet the images from these cameras
} do look better. Is it the bits and increased grey levels? I know that if
} your doing image analysis at the pixel level that more bits are to your
} advantage but what about just photgraphic quality?
}
} Rick Vaughn





From: Michael Plociniak :      plocinia-at-aecom.yu.edu
Date: Thu, 03 Feb 2000 12:44:23 -0500
Subject: New Gold Enhancement Reagents

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Dear Colleagues,
Has anyone used gold enhancement reagents from Nanoprobes, Inc.
(Stonybrook, NY - USA) as an alternative to silver enhancement for
enlarging 1 nm immunogold probes?

Advertisements state that gold enhancement has lower background and is
compatible with osmium. Can anyone verify this from personal experience?

In addition to these advantages, I am hoping that milder pH conditions will
be less damaging to ultrastructure in cultured neurons (using a
preembedment protocol).

Thank you,
Michael




From: Sinkler, Wharton :      wharton.sinkler-at-anlw.anl.gov
Date: Thu, 03 Feb 2000 12:13:10 -0700
Subject: Re: TEM:finding the parallel beam

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Rado,

} The diffraction plane is always the crossover plane not the theoretical back
} focal plane of the objective. Its z-position changes with the change of the
} C2 excitement.

Your statement above is not wrong, but isn't the standard TEM viewpoint (or
terminology). Yes, the plane at which the spot is sharpest (the in focus
plane for the demagnified filament image) is the crossover plane. However
to generally call this the "diffraction plane" is confusing. For example in
the case of CBED, the crossover plane has moved (up or down, depending on
whether C2 was initially under or overfocused) to the specimen plane. This
doesn't make the specimen plane the "diffraction plane". Rather, the
"diffraction plane" is now considered the in-focus plane for the C2
aperture. This is at the objective lens back-focal plane - the plane at
which points on the specimen are magnified to infinity and where the
position variable corresponds to the illumination angle. In collecting an
SADP you can correct for slightly non-parallel illumination by correcting
diffraction focus slightly off of the true back focal plane, but this should
be only slight.

What was proposed in the original mail was a technique of determining how
parallel the incident beam is, and of improving things by tweaking C2. If
the beam isn't very parallel, you'll see the diffraction spots shift when
you move the SA aperture. This is because the incident beam inclination
depends on the position on the sample.

I would think the "tweak" to C2 for obtaining more parallel illumination
would always in the direction of greater beam spread (unless I am
misunderstanding something?).

Regards,
Wharton
++++++++++++++++++++++++++++++++++++++++++++++++++
Wharton Sinkler
Argonne National Laboratory West
P. O. Box 2528
Idaho Falls, ID 83403



} Rado
}
} ---------------------------------------------------------------------
} Radostin Danev
} Laboratory of Ultrastructure Research
} National Institute for Physiological Sciences
} Myodaiji-cho, Okazaki 444-8585, JAPAN
} e-mail: rado-at-nips.ac.jp
} ---------------------------------------------------------------------
} ----- Original Message -----
} } From: Jonathan Barnard {Jonathan.Barnard-at-Angstrom.UU.SE}
} To: {microscopy-at-sparc5.microscopy.com}
} Sent: Wednesday, February 02, 2000 7:20 PM
} Subject: TEM:finding the parallel beam
}
}
} } ------------------------------------------------------------------------
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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} } -----------------------------------------------------------------------.
} }
} }
} } I got a tip for finding the condenser lens settings required for parallel
} } illumination from Angus Kirkland (Cambridge).
} }
} } Make sure you are in microprobe mode and set up for SAED and the specimen
} } is out of the way. Spread the beam, put in the SA Aperture, go to
} } diffraction and then sweep the SA aperture (SAA) from side to side and
} } minimise the deflection of the diffraction spot by tweaking the
} } illumination control (C2 or second condenser lens). A little bit of
} thought
} } will convince you that anything other than a parallel beam will give you a
} } side to side motion (tilt) when the SAA is swept.
} }
} } I have found it useful to keep a table of C2 condenser lens current
} } settings for each spot size (C1 excitation) in microprobe mode. Setting
} the
} } C2 lens for parallel illumination and adjusting the diffraction focus to
} } get the sharpest spot will then give you near perfect SAD conditions.
} }
} } Anyone contemplating electron holography for example, will have to start
} } from the parallel illumination to get the best spatial coherence for their
} } holograms.
} }
} } ********************************************************
} } Dr Jonathan Barnard
} }
} } Analytical Materials Physics
} } The Angstrom Laboratory, Uppsala University
} } P O Box 534, SE-751 21 Uppsala, Sweden
} } Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
} } http://www.angstrom.uu.se/analytical/home.html
} }
} } ********************************************************
} }
} }
}





From: Carl Henderson :      chender-at-umich.edu
Date: Thu, 3 Feb 2000 15:11:42 -0500
Subject: Re: Nitrogen determination

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Eugenia,

You can avoid your concern with C coating by coating your standards
and unknowns at the same time. You should use the minimum coating
thickness--just enough to prevent beam charging.

However, as stated below by Dr. Lachowski, ammonia volatility would
be a big problem, especially since you will need help from high beam
intensities and/or long counting times to get enough N counts to give
decent precision. The expected intensity measured with a 10kV beam
from a zeolite nominally of analcime composition (NaAlSi2O6.H2O) with
5 wt% N would be about 2.5% that of the intensity from Si3N4. High N
intensity is difficult to achieve even with the best spectrometer and
crystal designs. And to add insult to injury, the addition of the C
coating absorbs even more of the N x-rays from the sample. The mass
absorption coefficient for N K-alpha emission with a C absorber is
huge (greater than 23,000).

Plus, you should check for possible N peak shape differences between
Si3N4 (or other standards) and your ammonium-bearing zeolite.

All in all, it is a very difficult analytical problem.

Good luck!

Carl

At 4:24 PM +0000 2/3/00, Dr Eric Lachowski wrote:
} Eugenia,
} the microprobe is not the best tool for the analysis of
} ammonia in zeolites because ammonia is very volatile in the
} electron beam. If your zeolite is phase pure it would be
} far better to dissolve it and do a classical chemical
} analysis.
}
} regards,
} Eric
} On Thu, 3 Feb 2000 14:54:20 +0100 bengi {bengi-at-unimo.it}
} wrote:
}
} }
} } Dear listers,
} } I have started a research project on ammonium -bearing mineral (zeolites),
} } and now I'm trying to quantify N using electron microprobe analysis. My
} } problems of dealing with N are:
} }
} } a) The thickness of the coating (in the literateur =
} approximately 150 A).
} } Does anyone have information on the model of coater to making these films?
} } Does anyone have experience with preparation of such
} } samples?
} }
} } b) Obtaining appropriate standards. (I have Si3N4, but it contains too much
} } N (=39.96%) towards my samples (5%). Does anyone known any good nitrogen
} } standards for such a sample ?
} }
} } Any other suggestions about quantitative nitrogen analysis would be greatly
} } appreciated !!
} } Thanks for your time.
} }
} } My best regards
} }
} } Eugenia
} }
} }
} } Eugenia Marchi
} } Universit degli Studi di Modena
} } Dip. Scienze della Terra
} } P.le S.Eufemia n.19
} } 41100 Modena (Italy)
} } Tel. +39-059-417289
} } Fax. +39-059-417399
} } e-mail: bengi-at-unimo.it
} }
} }
} }
}
} ----------------------
} Dr Eric Lachowski
} Department of Chemistry
} University of Aberdeen
} Meston Walk
} Old Aberdeen AB24 3UE
} Scotland
} +44 (0)1224 272934 fax 272921
} e.lachowski-at-abdn.ac.uk

======================================
Carl Henderson
Electron Microbeam Analysis Laboratory
University of Michigan
2501 C.C. Little Bldg.
Ann Arbor, MI 48109-1063 USA
(734) 936-1550 FAX (734) 763-4690
======================================




From: Paul.Nolan-at-Alcan.Com
Date: Thu, 3 Feb 2000 16:05:19 -0600
Subject: Technoorg-Linda Ion Beam Thinner

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To: {Microscopy-at-MSA.Microscopy.Com}


Does anyone out there have experience with the Technoorg-Linda IV3H ion beam
thinner.
We have their instruction manual but it has been a long time since we had a
training session and are having some problem with the set up.
Does anyone have a revised instruction manual or perhaps some tips on the
alignment procedure of the guns. (please please please please).

Thanks
Paul Nolan

P.S..
It is cold in Canada..everyday...all year. hehe






From: Thomas, Larry :      Larry.Thomas-at-pnl.gov
Date: Thu, 03 Feb 2000 14:10:01 -0800
Subject: RE: TEM:finding the parallel beam

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Laurie,

Now I'm really confused. I used to think I understood this somewhat.

1. In the JEOL 2010F FEG (the "all bets are off" case in your item#7),
changing the illumination spread by changing C2 profoundly moves the
"diffraction plane" relative to the objective aperture. [There are actually two
objective apertures at different heights in this microscope, but that's beside
the point]. By diffraction plane, I mean the plane below the objective lens
where the diffraction spots are in sharp focus. In other words (assuming fixed
OL and condenser minilens excitations), in this microscope there is only one C2
value that allows the diffraction spot pattern and OL aperture to be in focus
simultaneously. If you focus on the OL aperture (with the intermediate lens),
the spot pattern can be focused at only this one C2 setting. At any other C2
setting, you can focus the diffraction pattern with the intermediate lens
("diffraction focus") but the OL aperture will then be defocused. Changing the
condenser minilens excitation changes the diffraction plane (relative to the OL
aperture) to a different C2 setting. This is actually quite a useful feature of
the microscope, e.g., for getting very intense focused diffraction patterns, but
a different story.

2. The reason for choosing to focus the diffraction spots rather than
the K-lines is --to put it simply-- that's where the intensity is. In amplitude
contrast imaging (conventional brightfield and darkfield imaging), the intensity
contribution to the image is limited by the OL aperture. It's really important
to have the defining aperture and diffraction spot pattern in focus
simultaneously. In addition, if I aim to measure diffraction spot patterns
there is little incentive to focus the K-lines instead of the spots.

3. Textbooks such as Hirsch et al. often discuss the positional error
in SA diffraction due to Cs. What readers often miss is the consequence of the
fact that the error depends strongly on the diffraction angle. If measurements
are confined to the low-index (low-angle) reflections, the selecting area can be
reduced (to 200 nm or so at 200 kV) without big positional errors. Maybe that's
an example of differences in experimental and theoretical viewpoints.

4. Reducing the condensor aperture size will give less illumination,
but it won't give parallel illumination. It will change the size of the
diffraction spots, but not their focus.

5. I'm not quite sure why the illumination spread has such a strong
effect on the apparent position of the diffraction plane relative to the OL
aperture, but suspect it arises from the action of the strong
condensor-objective in this microscope. I would also question that it has much
to do with coherent/incoherent beams in a FEG or with prefield/postfield Cs
effects.

Larry


Larry Thomas
Pacific Northwest National Laboratory
MSIN P8-16
P.O. Box 999
Richland, WA 99352
Phone: (509)372-0793 Fax: (509)376-6308
Email: mailto: Larry.Thomas-at-pnl.gov



----------
From: L. D. Marks
Reply To: L-marks-at-nwu.edu
Sent: Thursday, February 3, 2000 8:06 AM
To: Radostin Danev
Cc: MSA listserver
Subject: Re: TEM:finding the parallel beam

------------------------------------------------------------------------
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
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Sorry, but I think we are getting a little confused here.

1) The diffraction plane is (by definition) the objective lens
focal length below the sample - it is the focus of parallel
directions coming from the sample. The focal length (and
parameters such as Cs, Cc) change rapidly with the objective lens
current so an eucentric position makes life simple but is not
necessarily the best condition to use. (For HREM you generaly
want to have the objective lens stronger and reduce Cs.)

2) In general C2 does not change the focal length at all. You
can have some effect from coupling of the magnetic fields, but
all C2 is supposed to do is change the convergence angle.

3) The objective aperature is (approximately) in the back-focal
plane, given that it is finite and height adjustments are only
done coarsely (for a specific value of the objective current
only).

4) The selected area aperature is (approximately) in the same
plane as the object for the same reasons as above. Remember that
there are positional errors in SA diffraction due to Cs (see
Hirsch et al for instance).

5) You should focus Kikuchi-lines, not go for the smallest spot,
since these are real object in the diffraction pattern. What you
will see is then representative of you incident beam, sometimes
a Gaussian range of directions.

6) The simplest way to get more parallel illumination in general is
to reduce the condensor aperture size.

7) All bets are off if you have a FEG. Rather than having incoherent
illumination things like the Cs of the prefield (above the sample)
and postfield (below) add coherently making it much more complicated.
For instance, the illumination angle varies across the field of view.
While this is also there with LaB6, it is a weak effect (except for
certain classes of diffracted beams, e.g. those with a screw-axis).
With
a FEG you can get a "spot" pattern many ways, and most of these will
have
large aberrations (e.g. pincushion).

++++++++++++++++++++++++++++++++++++++++++++++++
Laurence Marks
Department of Materials Science and Engineering
Northwestern University
fax: (847) 491-7820
mailto:l-marks-at-nwu.edu
http://www.numis.nwu.edu
++++++++++++++++++++++++++++++++++++++++++++++++






From: David Henriks :      Henriks-at-CompuServe.COM
Date: Thu, 3 Feb 2000 17:57:35 -0500
Subject: Technoorg-Linda Ion Beam Thinner

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Paul:

South Bay Technology does have quite a bit of experience with the TL IV3
Ion Mill and we are happy to offer our assistance. We market the IV3
system throughout the world for Technoorg-Linda and offer technical support
as well. I will send you the most up to date IV3 manual that we have
produced and will include by separate e-mail the section on the gun
alignment.

I will follow up with you to be sure that you have all of the information
you need.

Best regards-

David
Writing at 3:50:09 PM on 02/03/2000

***************************************************************************
************************

David Henriks
Vice President TEL: 800-728-2233
(toll free in the USA)
South Bay Technology, Inc. +1-949-492-2600
1120 Via Callejon FAX: +1-949-492-1499
San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com

***************************************************************************
************************

} } } } } Please visit us at http://www.southbaytech.com { { { { {

Manufacturers of precision sample preparation equipment and supplies for
metallography, crystallography and electron microscopy.

Message text written by
INTERNET:"Paul.Nolan-at-Alcan.Com"-at-sparc5.Microscopy.Com
}
------------------------------------------------------------------------
The Microscopy ListServer -- Sponsor: The Microscopy Society of America


Does anyone out there have experience with the Technoorg-Linda IV3H ion
beam
thinner.
We have their instruction manual but it has been a long time since we had
a
training session and are having some problem with the set up.
Does anyone have a revised instruction manual or perhaps some tips on the
alignment procedure of the guns. (please please please please).

Thanks
Paul Nolan

P.S..
It is cold in Canada..everyday...all year. hehe
{





From: L. D. Marks :      ldm-at-risc4.numis.nwu.edu
Date: Thu, 3 Feb 2000 17:08:04 -0600 (CST)
Subject: RE: TEM:finding the parallel beam

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


On Thu, 3 Feb 2000, Thomas, Larry wrote:

} Laurie,
}
} Now I'm really confused. I used to think I understood this somewhat.
}
} 1. In the JEOL 2010F FEG (the "all bets are off" case in your item#7),
} changing the illumination spread by changing C2 profoundly moves the
} "diffraction plane" relative to the objective aperture. [There are actually two
} objective apertures at different heights in this microscope, but that's beside
} the point]. By diffraction plane, I mean the plane below the objective lens
} where the diffraction spots are in sharp focus. In other words (assuming fixed
} OL and condenser minilens excitations), in this microscope there is only one C2
} value that allows the diffraction spot pattern and OL aperture to be in focus
} simultaneously. If you focus on the OL aperture (with the intermediate lens),
} the spot pattern can be focused at only this one C2 setting. At any other C2
} setting, you can focus the diffraction pattern with the intermediate lens
} ("diffraction focus") but the OL aperture will then be defocused. Changing the
} condenser minilens excitation changes the diffraction plane (relative to the OL
} aperture) to a different C2 setting. This is actually quite a useful feature of
} the microscope, e.g., for getting very intense focused diffraction patterns, but
} a different story.
}
OK, the bets off case. In a FEG you have a somewhat coherent
range of incident directions. The "diffraction pattern" is therefore
something which is effected by the coherent aberrations of the microscope,
unlike the case with incoherent illumination. Consider the case with
focussed illumination, when the diffraction pattern shows an image of
the condensor aperture. In a FEG As a you can "focus" this (coherent)
image to a small spot by going out of focus in a true sense for the
diffraction pattern. This gives you a spot-like pattern, but you will also
see severe distortions at higher angles. With incoherent illumination you
cannot do this and going out of focus (in the diffraction pattern) will
give in general a blurred image of the condensor aperture - life is
simple.
What you need to do is focus "real diffraction" features, e.g.
Kikuchi lines, then not play with the diffraction focus at all. As you
change C2 the size of the spots will grow or shrink, this is fine.

} 2. The reason for choosing to focus the diffraction spots rather than
} the K-lines is --to put it simply-- that's where the intensity is. In amplitude
} contrast imaging (conventional brightfield and darkfield imaging), the intensity
} contribution to the image is limited by the OL aperture. It's really important
} to have the defining aperture and diffraction spot pattern in focus
} simultaneously. In addition, if I aim to measure diffraction spot patterns
} there is little incentive to focus the K-lines instead of the spots.
}
Yes, you can always get a "spot" pattern with a FEG, and it may
be prettier. However, beware the distortions which make doing
measurements from it very dangerous.

} 3. Textbooks such as Hirsch et al. often discuss the positional error
} in SA diffraction due to Cs. What readers often miss is the consequence of the
} fact that the error depends strongly on the diffraction angle. If measurements
} are confined to the low-index (low-angle) reflections, the selecting area can be
} reduced (to 200 nm or so at 200 kV) without big positional errors. Maybe that's
} an example of differences in experimental and theoretical viewpoints.
}
Hirsch et al is just a good reference to read, about this as well
as many other things!

} 4. Reducing the condensor aperture size will give less illumination,
} but it won't give parallel illumination. It will change the size of the
} diffraction spots, but not their focus.
}
It makes it closer to parallel. Of course, with really parallel
illumination there is no intensity!

} 5. I'm not quite sure why the illumination spread has such a strong
} effect on the apparent position of the diffraction plane relative to the OL
} aperture, but suspect it arises from the action of the strong
} condensor-objective in this microscope. I would also question that it has much
} to do with coherent/incoherent beams in a FEG or with prefield/postfield Cs
} effects.
}
Sorry, I don't think that is correct. I would suggested that
people look at the distortions with a standard sample and see how bad they
can be.

} Larry
}
}
} Larry Thomas
} Pacific Northwest National Laboratory
} MSIN P8-16
} P.O. Box 999
} Richland, WA 99352
} Phone: (509)372-0793 Fax: (509)376-6308
} Email: mailto: Larry.Thomas-at-pnl.gov
}
}
}
} ----------
} From: L. D. Marks
} Reply To: L-marks-at-nwu.edu
} Sent: Thursday, February 3, 2000 8:06 AM
} To: Radostin Danev
} Cc: MSA listserver
} Subject: Re: TEM:finding the parallel beam
}
} ------------------------------------------------------------------------
} 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
} -----------------------------------------------------------------------.
}
}
} Sorry, but I think we are getting a little confused here.
}
} 1) The diffraction plane is (by definition) the objective lens
} focal length below the sample - it is the focus of parallel
} directions coming from the sample. The focal length (and
} parameters such as Cs, Cc) change rapidly with the objective lens
} current so an eucentric position makes life simple but is not
} necessarily the best condition to use. (For HREM you generaly
} want to have the objective lens stronger and reduce Cs.)
}
} 2) In general C2 does not change the focal length at all. You
} can have some effect from coupling of the magnetic fields, but
} all C2 is supposed to do is change the convergence angle.
}
} 3) The objective aperature is (approximately) in the back-focal
} plane, given that it is finite and height adjustments are only
} done coarsely (for a specific value of the objective current
} only).
}
} 4) The selected area aperature is (approximately) in the same
} plane as the object for the same reasons as above. Remember that
} there are positional errors in SA diffraction due to Cs (see
} Hirsch et al for instance).
}
} 5) You should focus Kikuchi-lines, not go for the smallest spot,
} since these are real object in the diffraction pattern. What you
} will see is then representative of you incident beam, sometimes
} a Gaussian range of directions.
}
} 6) The simplest way to get more parallel illumination in general is
} to reduce the condensor aperture size.
}
} 7) All bets are off if you have a FEG. Rather than having incoherent
} illumination things like the Cs of the prefield (above the sample)
} and postfield (below) add coherently making it much more complicated.
} For instance, the illumination angle varies across the field of view.
} While this is also there with LaB6, it is a weak effect (except for
} certain classes of diffracted beams, e.g. those with a screw-axis).
} With
} a FEG you can get a "spot" pattern many ways, and most of these will
} have
} large aberrations (e.g. pincushion).
}
} ++++++++++++++++++++++++++++++++++++++++++++++++
} Laurence Marks
} Department of Materials Science and Engineering
} Northwestern University
} fax: (847) 491-7820
} mailto:l-marks-at-nwu.edu
} http://www.numis.nwu.edu
} ++++++++++++++++++++++++++++++++++++++++++++++++
}
}
}

++++++++++++++++++++++++++++++++++++++++++++++++
Laurence Marks
Department of Materials Science and Engineering
Northwestern University
fax: (847) 491-7820
mailto:L-marks-at-nwu.edu
http://www.numis.nwu.edu
++++++++++++++++++++++++++++++++++++++++++++++++






From: Tobias Baskin :      BaskinT-at-missouri.edu
Date: Fri, 4 Feb 2000 12:30:27 +1100
Subject: Re: LSCM Need help with non-fluorescing resin

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Greetings,
Butyl methyl methacrylate is in our hands non fluorescent
totally. You can use it for TEM, although it is not as good as epoxies.
Hope this helps.
Tobias
}
}
} I am working on fluorescent imaging of thin sections cut from samples
} embedded in TEM embedding resin, and am having a significant problem with
} resin fluorescence. Does anyone know of an embedding resin that doesn't
} autofluoresce?
}
} Matt Plantinga
} Amway Corporation
} matt_plantinga-at-amway.com





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From: ctschristopher :      ctschristopher-at-samiot.uct.ac.za (by way of Carol
Date: Thu, 3 Feb 2000 17:56:46 -0800
Subject: a question about 2-hydroxyhexane-dial, AKA hydroxyadipaldehyde

Contents Retrieved from Microscopy Listserver Archives
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LIst,

Does anyone know of a current source for this chemical? It's an ingredient
of an embedding medium made up by our group. We have a supply on hand but
have to reserve it if it's no longer commercially available.


} } We are very excited about the possibilities of HACH as a solution for a
} } problem
} } that has troubled us for a couple of years with our PU grafts. We have,
} } unfortunately, one serious problem in that we cannot find a source for HA.
} } We have tried Sigma Aldrich and they informed us that they withdrew
} } 2-Hydroxyhexane-dial from their range in 1990. We have tried a web search
} } and are contacting our local suppliers, so far without success. Do you have
} } any ideas where we can find it? We are very anxious to try HACH on our PU
} } samples and will gratefully appreciate any additional help that you can give
} } us.

} } Thanks again

} } Phil






From: Dr Malcolm Roberts :      malc-at-rock.ru.ac.za
Date: Fri, 04 Feb 2000 09:04:34 +0200
Subject: Silence

Contents Retrieved from Microscopy Listserver Archives
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Have I been cut off, or is nobody talking anymore?

--
Dr MP Roberts Phone: [+27](0)46 603 8313
Dept of Geology Fax: [+27](0)46 622 9715
Rhodes University e-mail: malc-at-rock.ru.ac.za
6140 Grahamstown
SOUTH AFRICA






From: Alexander Mironov Jr. :      amironov-at-cmns.mnegri.it
Date: Fri, 4 Feb 2000 08:51:22 +0100 (MET)
Subject: Re: New Gold Enhancement Reagents

Contents Retrieved from Microscopy Listserver Archives
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Dear Michael,

I have tried GoldEnhance for preembedding protocol to label intracellular
structures. It is marvelous. Buy it and use it - you will not be
unsatisfied. I have no any interest in Nanoprobes, I just like these dense
gold particles.
Practically all they claim is true:
- light insensitive
- no self-nucleation
- do not react with buffer ions
- is not dissolved by uranil and osmium (I have treat for 1 h)

Sincerely,
Dr. Alexander A. Mironov Jr., MD, PhD
Unit of Morphology
Dept. of Cell Biology and Oncology
Consorzio Mario Negri Sud
Via Nazionale, S.Maria Imbaro (Ch)
66030 Italy

Tel. 0039-0872-570-332
Fax 0039-0872-570-412
E-mail: amironov-at-cmns.mnegri.it


On Thu, 3 Feb 2000, Michael Plociniak 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 Colleagues,
} Has anyone used gold enhancement reagents from Nanoprobes, Inc.
} (Stonybrook, NY - USA) as an alternative to silver enhancement for
} enlarging 1 nm immunogold probes?
}
} Advertisements state that gold enhancement has lower background and is
} compatible with osmium. Can anyone verify this from personal experience?
}
} In addition to these advantages, I am hoping that milder pH conditions will
} be less damaging to ultrastructure in cultured neurons (using a
} preembedment protocol).
}
} Thank you,
} Michael
}
}





From: Ken Tiekotter :      tiekotte-at-up.edu
Date: Fri, 4 Feb 2000 00:13:57 -0800 (PST)
Subject: Re: Help:A comparison of optics

Contents Retrieved from Microscopy Listserver Archives
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Dear Steve and interested parties:

The comparison between Zeiss and Olympus '100x' objective lenses is
impossible unless one knows the type of lens be compared.

Catagories used to determine the quality of such a lens include:
aberration correction (spherical and chromatic), correction of flatness of
field, numerical aperture, the presence of an iris diaphram, whether the
system is based on a 160mm tube length or infinity correction, and if the
lens is used for brightfield or phase contrast for instance.

(PLAN APO, PH3, 100x, iris diaphram, 1.32 numerical aperture? 160mm tube
length)

Resolution is still basically related to half the wavelength utilized.
Hey, let's look at the advantages of oblique illumination for contrast
enhancement and a differerent formula for resolution!!

Is your client looking at a system or an indivdual lens? Do they have a
Zeiss or an Olympus system? One does not mixed and match objectives from
different systems merely for the convenience of pricing.

An additional consideration is the type and correction of the condenser
lens. Check the numerical aperture of not only the objective, but the
condenser lens as well.

More questions than answers...Let me know if I can further complicate the
answers!

Cheers!
Ken

------------
Ken Tiekotter
Dept. of Biology
The University of Portland
5000 N Willamette Blvd.
Portland, OR 97303


On Thu, 3 Feb 2000, Steve Niemela wrote:

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} -----------------------------------------------------------------------.
}
}
} To: {Microscopy-at-MSA.Microscopy.Com}
} Subject: A comparison of optics
} Date: Thu, 3 Feb 2000 10:57:19 -0600
} MIME-Version: 1.0
} Content-Type: multipart/alternative;
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} X-Mailer: Microsoft Outlook Express 5.00.2615.200
} X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2615.200
}
} A client of ours wonders about a comparison between Zeiss and Olympus
} optics. The Zeiss 100X lense cost $10,000, and the Olympus about $4,000:
} does that mean the Olympus is inferior?  That's what our client's
} colleagues imply. What measure of lense function is relevant? Does Olympus
} (or Zeiss) have a specification, like 'resolves particles to 0.1 micron'
} or something like that?  Is there a measure of distortion? Best
} regards, Steve NiemelaVayTek, Inc.305 West Lowe AvenueFairfield, IA
} 52556email: sniemela-at-vaytek.com Phone: 515.472.2227Fax: 515.472.8131web:
} www.vaytek.comgeneral email: vaytek-at-vaytek.com
}
}
}
}





From: Janko Otto :      otto-at-quantifoil.com
Date: Fri, 04 Feb 2000 09:47:44 +0100
Subject: Re: JAMP-7810 manual in English and local chemical analysis

Contents Retrieved from Microscopy Listserver Archives
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Dear Dmitri,

} We are making preparations for experiments on determination of the chemical
} composition of semiconductor oxides, formed locally. The size of the
} modified area should be about 1 micron, and we are not sure yet if it is
} possible to make mentioned chemical analysis with Auger microprobe at all.

I can recommend you some former colleagues of mine performing auger
spectroscopy and many other semiconductor surface analytics.
Please contact Prof. Wolfgang Richter: richter-at-pinet.uni-jena.de.
The URL is: http://www.physik.uni-jena.de/~layer/

Regards,
Kay Pfennighaus


--
_____________________________________________________________

Janko Otto/Kay Pfennighaus email otto-at-quantifoil.com
Quantifoil Micro Tools GmbH Tel +49 (0) 3641 - 206 470
Winzerlaer Strasse 10 Fax +49 (0) 3641 - 206 471
D-07745 Jena Web http://www.quantifoil.com
Germany
_____________________________________________________________




From: Tony Bruton :      bruton-at-EMU.UNP.AC.ZA
Date: Fri, 04 Feb 2000 11:38:49 +0200
Subject: Re: Silence

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


W'ere all waiting to hear news of MSSA Conference 2000 { :-)

Tony

} } } Dr Malcolm Roberts {malc-at-rock.ru.ac.za} 02/04/00 09:04AM } } }
------------------------------------------------------------------------
The Microscopy ListServer -- Sponsor: The Microscopy Society of America


Have I been cut off, or is nobody talking anymore?

--
Dr MP Roberts Phone: [+27](0)46 603 8313
Dept of Geology Fax: [+27](0)46 622 9715
Rhodes University e-mail: malc-at-rock.ru.ac.za
6140 Grahamstown
SOUTH AFRICA








From: A.Walker :      Alan.Walker-at-sheffield.ac.uk
Date: Fri, 4 Feb 2000 10:10:23 +0000
Subject: Re: Technoorg-Linda Ion Beam Thinner

Contents Retrieved from Microscopy Listserver Archives
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Hi Paul,

} Does anyone out there have experience with the Technoorg-Linda IV3H ion beam
} thinner.
} We have their instruction manual but it has been a long time since we had a
} training session and are having some problem with the set up.
} Does anyone have a revised instruction manual or perhaps some tips on the
} alignment procedure of the guns. (please please please please).

We have the IV3 here in Sheffield - I'll try and help you if I can.
You need to be able to actually see the beams (cover your head and
the viewing glass with a black sheet if you cannot darken the room).
Get one gun to fire through the center of the sample holder (tilt it
at right angles to the beam) and to hit the opposite gun where it's
beam would exit. Do the same for the other gun. Now keep this gun
orientation, load the specimen and tilt the holder to the desired milling angle.

Hope this helps,

Alan Walker

*********************************************
Alan Walker
Dept of Electronic and Electrical Engineering
University of Sheffield
Mappin Street
Sheffield S1 3JD
United Kingdom

Tel:+44-(0)114-2225365
Fax:+44-(0)114-2726391
alan.walker-at-shef.ac.uk
http://www.shef.ac.uk/uni/academic/D-H/eee/
*********************************************




From: Donald Lovett :      lovett-at-tcnj.edu
Date: Fri, 4 Feb 2000 09:17:35 -0500 (EST)
Subject: Is KEVEX 8000 worth installing?

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A Kevex 8000 (with huge 8 or 10" disk drives), attached to an old Amray
1000, has been donated to us. All components were fully functional when
they were crated up a year ago. (They are sitting in our basement until we
move into our new building this June).

A colleague in Physics (I'm a biologist) wishes to use the system to
quantify elemental composition (beryllium, gallium, etc) in semi-conductor
ceramics. We already have (up and running) an Hitachi HS-510 (SEM) and an
Hitachi H-7100 (STEM).

1) Is this (Kevex 8000) an appropriate instrument for her to do the
analyses?

2) Which would be wiser: Trying to get the Amray installed and running
or getting an adapter to use the Kevex in one of the Hitachi's? (Space is
a consideration, so I'd prefer not to have to install (and keep in running
condition) another scope.)

3) The Kevex is pretty old. Are we wasting money trying to get or keep
running something that is this old? I realize that we may need to replace
the detector, since it has been at room temp for about a year. Is there
anything else we should automatically consider replacing or upgrading as
part of the installation?

4) She also has been sending her samples out to do x-ray analysis of the
material to determine it's crystalline structure. The place she sends it
to has a special x-ray diffraction instrument. Could the x-ray
diffraction on the H-7100 be used to get equal results, or does this other
instrument have capabilities that the TEM doesn't.

Thanks for any advice.

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










From: Barbara Foster :      mme-at-map.com
Date: Fri, 04 Feb 2000 10:20:50 -0500
Subject: Re: Help:A comparison of optics

Contents Retrieved from Microscopy Listserver Archives
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Steve,

The cost of an objective depends on a number of issues, not the least of
which are the corrections for chromatic and spherical aberration as well as
the size of the numerical aperture. I'd need more information to really
compare. "Optimizing Light Microscopy" has a detailed discussion of all of
this which might be helpful. Ordering information is available on our
website.

The best test is to try each system with your applications. Since much of
Vaytek's work centers on deconvolution of fluorescence images, fluorescent
beads of varying sizes would be a good test object for you and high
throughput and crisp imaging would probably be two key benchmarks. Since
fluorescence detects objects beyond the resolution limits, the normal
resolution tests are meaningless. By the way, the numerical aperture of
the objective is the major deciding factor on a microscope's ability to
resolve fine detail (as well as providing good edge information).

If you'd like to write or call me directly, I may be able to give you
further guidance.

Best regards,
Barbara Foster
Consortium President
Microscopy/Microscopy Education ...Educating microscopists for greater
productivity.

125 Paridon Street Suite 102 Springfield, MA 01118
PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Visit our web site {http://www.MME-Microscopy.com/education}


Caveat: MME does have a commercial interest in the sale of "Optimizing
Light MIcroscopy:"

At 04:04 PM 2/3/00 -0600, Steve Niemela wrote:
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From: Warren E Straszheim :      wesaia-at-iastate.edu
Date: Fri, 04 Feb 2000 09:52:32 -0600
Subject: Re: Is KEVEX 8000 worth installing?

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If your Kevex has a Quantum thin window detector you should be able to see
about half of a Boron peak, but there is no commercially available detector
(that I know of) that reliably sees all of the beryllium peak. And if you
have a standard window, you will only be able to see sodium and above.

I suppose the detector may still be functional even after storage. I recall
that it is not a trivial thing to switch detectors between scope models.
The adapters can be quite different and can get pricey. But I have not
priced one in over 10 years.

I suppose with rigorous use of standards, you might be able to analyze Be
by difference, but it would be a trick.

I think the Kevex 8000 may be worth installing if you are primarily
interested in x-ray analysis. We used a Kevex Delta for many years and
found it quite adequate. I might be able to talk you through some of the
technical issues.

However, if you are interested in digital imaging or x-ray mapping, the
improvements in the new equipment is fantastic compared to the old systems.
We run an IXRF Systems EDS on a JEOL 840A (having replaced our Kevex) and a
Link ISIS on a Hitachi 2460N. I could hardly imagine giving up their
capabilities and going back to the Delta or an 8000. Disk storage, the
operating environment, digital imaging can hardly be compared between the
old and the new.

FWIW,
Warren

At 09:17 AM 2/4/2000 -0500, Donald L. Lovett wrote:
} A Kevex 8000 (with huge 8 or 10" disk drives), attached to an old Amray
} 1000, has been donated to us. All components were fully functional when
} they were crated up a year ago. (They are sitting in our basement until we
} move into our new building this June).
}
} A colleague in Physics (I'm a biologist) wishes to use the system to
} quantify elemental composition (beryllium, gallium, etc) in semi-conductor
} ceramics. We already have (up and running) an Hitachi HS-510 (SEM) and an
} Hitachi H-7100 (STEM).
}
} 1) Is this (Kevex 8000) an appropriate instrument for her to do the
} analyses?
}
} 2) Which would be wiser: Trying to get the Amray installed and running
} or getting an adapter to use the Kevex in one of the Hitachi's? (Space is
} a consideration, so I'd prefer not to have to install (and keep in running
} condition) another scope.)
}
} 3) The Kevex is pretty old. Are we wasting money trying to get or keep
} running something that is this old? I realize that we may need to replace
} the detector, since it has been at room temp for about a year. Is there
} anything else we should automatically consider replacing or upgrading as
} part of the installation?
}
} 4) She also has been sending her samples out to do x-ray analysis of the
} material to determine it's crystalline structure. The place she sends it
} to has a special x-ray diffraction instrument. Could the x-ray
} diffraction on the H-7100 be used to get equal results, or does this other
} instrument have capabilities that the TEM doesn't.
}
} Thanks for any advice.
}
} Don





From: Jim McGee :      mcgee-at-geol.sc.edu
Date: Fri, 04 Feb 2000 10:55:56 -0500
Subject: Re: Nitrogen determination

Contents Retrieved from Microscopy Listserver Archives
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Carl et al.,

I responded to Eugenia's query on another listserver (MAS microprobe listserver),
but since it is also discussed here, I will add a couple things.

I did some work on ammonium-bearing minerals (with ~ 3 - 5 wt%N), using BN and later
synthetic buddingtonite (ammonium feldspar) as the N standard (see Microbeam
Analysis 1994, New Orleans meeting). I avoided using any epoxy for the sample prep
as a precaution against N artifacts from the mounting medium. The synthetic
buddingtonite can be made ala Voncken et al., 1988 (Phys. Chem of Minerals, v. 15,
p. 323).

We did "normal" carbon coating, using brass color as a guide to obtain 150-200A
coating (standards and unknowns). I used a synthetic crystal, 2d 60A, and ran at
10kV, 25 nA probe current. I agree that a light C coat, done at same time/conditions
as the standards is important. I don't recall a substantial problem with mobility
at thnese operating conditions, although the N intensity yields were low and
detection limits were on the order of 0.5wt% N. The minerals are feldpar and
perhaps the ammonium ion stayed put better than it woud in zeolites. I agree
though, it is a difficult analytical problem. But that makes it interesting.

Jim
--
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
James J. McGee (email: jmcgee-at-sc.edu)
Department of Geological Sciences
University of South Carolina
Columbia, SC 29208

Tel: 803-777-6300 Fax: 803-777-6610




Carl Henderson wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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} -----------------------------------------------------------------------.
}
} Eugenia,
}
} You can avoid your concern with C coating by coating your standards
} and unknowns at the same time. You should use the minimum coating
} thickness--just enough to prevent beam charging.
}
} However, as stated below by Dr. Lachowski, ammonia volatility would
} be a big problem, especially since you will need help from high beam
} intensities and/or long counting times to get enough N counts to give
} decent precision. The expected intensity measured with a 10kV beam
} from a zeolite nominally of analcime composition (NaAlSi2O6.H2O) with
} 5 wt% N would be about 2.5% that of the intensity from Si3N4. High N
} intensity is difficult to achieve even with the best spectrometer and
} crystal designs. And to add insult to injury, the addition of the C
} coating absorbs even more of the N x-rays from the sample. The mass
} absorption coefficient for N K-alpha emission with a C absorber is
} huge (greater than 23,000).
}
} Plus, you should check for possible N peak shape differences between
} Si3N4 (or other standards) and your ammonium-bearing zeolite.
}
} All in all, it is a very difficult analytical problem.
}
} Good luck!
}
} Carl
}
} At 4:24 PM +0000 2/3/00, Dr Eric Lachowski wrote:
} } Eugenia,
} } the microprobe is not the best tool for the analysis of
} } ammonia in zeolites because ammonia is very volatile in the
} } electron beam. If your zeolite is phase pure it would be
} } far better to dissolve it and do a classical chemical
} } analysis.
} }
} } regards,
} } Eric
} } On Thu, 3 Feb 2000 14:54:20 +0100 bengi {bengi-at-unimo.it}
} } wrote:
} }
} } }
} } } Dear listers,
} } } I have started a research project on ammonium -bearing mineral (zeolites),
} } } and now I'm trying to quantify N using electron microprobe analysis. My
} } } problems of dealing with N are:
} } }
} } } a) The thickness of the coating (in the literateur =
} } approximately 150 A).
} } } Does anyone have information on the model of coater to making these films?
} } } Does anyone have experience with preparation of such
} } } samples?
} } }
} } } b) Obtaining appropriate standards. (I have Si3N4, but it contains too much
} } } N (=39.96%) towards my samples (5%). Does anyone known any good nitrogen
} } } standards for such a sample ?
} } }
} } } Any other suggestions about quantitative nitrogen analysis would be greatly
} } } appreciated !!
} } } Thanks for your time.
} } }
} } } My best regards
} } }
} } } Eugenia
} } }
} } }
} } } Eugenia Marchi
} } } Universit degli Studi di Modena
} } } Dip. Scienze della Terra
} } } P.le S.Eufemia n.19
} } } 41100 Modena (Italy)
} } } Tel. +39-059-417289
} } } Fax. +39-059-417399
} } } e-mail: bengi-at-unimo.it
} } }
} } }
} } }
} }
} } ----------------------
} } Dr Eric Lachowski
} } Department of Chemistry
} } University of Aberdeen
} } Meston Walk
} } Old Aberdeen AB24 3UE
} } Scotland
} } +44 (0)1224 272934 fax 272921
} } e.lachowski-at-abdn.ac.uk
}
} ======================================
} Carl Henderson
} Electron Microbeam Analysis Laboratory
} University of Michigan
} 2501 C.C. Little Bldg.
} Ann Arbor, MI 48109-1063 USA
} (734) 936-1550 FAX (734) 763-4690
} ======================================





From: Augusto_A_Morrone-at-notes.seagate.com
Date: Fri, 4 Feb 2000 09:58:13 -0600
Subject: JEOL 2010 question/C2, diffraction and OA

Contents Retrieved from Microscopy Listserver Archives
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Since the discussion on CM12 alignment shifted to several other problems, and
eventually got some JEOL 2010 users involved (Larry Thomas), I thought of
throwing in this alignment question regarding the OA in a 2010 with a LaB6 gun:

To keep the camera length at a consistent value when I am not using an internal
standard, I record SA diffraction patterns with C2 fully defocused cw (this is
by conventional jargon, although actually using the Brightness knob I am
changing C3). This is supposed to make the illumination as parallel as possible
and is easily reproducible. Then I focus the direct beam with the intermediate
lens (diffraction focus). CBD and NBD are another problem.

The problem is to align the Objective Aperture under this condition. If the OA
is centered around the direct beam in diffraction, when switching to image mode
and increasing the illumination (Brightness) for imaging conditions, the
aperture will be apparently out of center. This gives the impression that the
voltage center changes between a fully defocused C2 and a moderately focused
(still cw from crossover, converging the illumination on the sample) condenser.
That shift is obvious in DIFF mode, the direct beam can be seen to shift as C2
is turned ccw from the fully cw position.

As was noted in the on-going discussion, the crossover planes move along the
optic axis as the strength of C2 changes. The helicoidal path of the beam is
straight down the optic axis of the lenses only piecewise, and at the level of
the OA plane it stays centered only for a limited range of C2 settings--or
convergence angles. In my case, the convergence changes considerably when I
record images digitally in a 1kx1k CCD camera, which screams for brightness.

My two ways around this are to (1) recenter the OA in image mode, or (2) in DIFF
mode, but keeping C2 at the setting I will use in MAG mode. Both methods are a
pain, mostly when doing BF/DF work, and probably incorrect.

Is there an alignment procedure for the 2010 that will prevent this " OA shift "
from DIFF to MAG modes?

Augusto Morrone
Seagate Technology
NRW-115
7801 Computer Ave S.
(612) 844-5838
Fax: (612) 844-7301






From: Mary Mager :      mager-at-interchange.ubc.ca
Date: Fri, 04 Feb 2000 08:42:59 -0800
Subject: Re: Need repairman for Hitachi SEM

Contents Retrieved from Microscopy Listserver Archives
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Dear Paul,
Why don't you use the Hitachi service from NSC? They are the best and the
cheapest.
At 12:12 PM 2/3/00 -0500, you wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America


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





From: Lesley Weston :      lesley-at-interchange.ubc.ca
Date: Fri, 4 Feb 2000 10:12:49 -0800 (PST)
Subject: Aqueous mounting medium

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Can anyone recommend an aqueous mounting medium that polymerises (or
whatever) to form a permanent hard mount, and that does not
autofluoresce? Thanks.

Lesley Weston.







From: Douglas Keene :      DRK-at-SHCC.ORG
Date: Fri, 04 Feb 2000 10:36:37 -0800 (Pacific Standard Time)
Subject: New Gold Enhancement Reagents

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Michael,

We had been using silver enhancement kits from Amersham and
Nanoprobe. Of these, we found the Nanoprobe kit to be very
difficult to work with given it's high viscosity. We also
use a pre-embed protocol, and found that both silver
enhancement kits caused collagen fibrils to denature
(unravel). However, during our protocol the tissue is
incubated for fairly long duration in the enhancement
medium (15 minutes on ice, then warmed to 25 C in a water
bath for an additional 5 minutes, all in the dark).

Our more recent experience is with the Nanoprobe gold
enhancement kit. It has HUGE advantages over the silver
system.

First, you can Osmicate the tissue. Second, the viscosity
of the components is comparatively very low, easing the use
of the product. It is not so sensitive to light, and it
does not cause denaturation of collagen fibrils. We do see
some variability in particulate size, but this is probably
a problem associated with diffusion of the media through
the tissue. We have found no disadvantages of the Gold
enhancement v.s. Silver and now use the Gold method
exclusively.

I hope this helps,

Doug

On Thu, 03 Feb 2000 12:44:23 -0500 Michael Plociniak
{plocinia-at-aecom.yu.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
} -----------------------------------------------------------------------.
}
}
} Dear Colleagues,
} Has anyone used gold enhancement reagents from Nanoprobes,
} Inc. (Stonybrook, NY - USA) as an alternative to silver
} enhancement for enlarging 1 nm immunogold probes?
}
} Advertisements state that gold enhancement has lower
} background and is compatible with osmium. Can anyone
} verify this from personal experience?
}
} In addition to these advantages, I am hoping that milder pH
} conditions will be less damaging to ultrastructure in
} cultured neurons (using a preembedment protocol).
}
} Thank you,
} Michael

----------------------
Douglas R. Keene
Associate Investigator
Shriners Hospital Research Facilities
3101 S.W. Sam Jackson Park Road
Portland, Oregon 97201
phone: 503-221-3434
FAX: 503-412-6894 (9-5 PST)
e-mail: DRK-at-shcc.org








From: Dan Freidus :      freidus-at-wwnet.com
Date: Fri, 4 Feb 2000 14:38:50 -0500
Subject: Re: Help:A comparison of optics

Contents Retrieved from Microscopy Listserver Archives
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-----Original Message-----
} From: Ken Tiekotter {tiekotte-at-up.edu}
..One does not mixed and match objectives from different systems merely for
the convenience of pricing....
----------------------------------------------------------------------------
---------------------------------------------------------------------------

Of course, I've heard this sort of statement many times before. But has
anyone ever done any comparisons of mixed systems to see if there are
certain brands that are particularly incompatible or particularly
compatible?

I ask this both as a general question and because I use a Wild M7 for which
I need a pair of high eyepoint oculars. I don't need the adjusting collar
built into current models and haven't been able to find an older pair used.
But before I buy a Nikon/Olympus/ etc. pair I'd love to know how different
the complete system will be from what I've got now.

I know many people who have used third party photo eyepieces, in just the
situation where you'd think quality would be of the utmost concern. Dealers
are often not the best source of info since they tend to represent one brand
and, even when they are completely honest, rarely have experienc ewith a
wide range of brands (especially mixed up.

Dan Freidus
freidus-at-wwnet.com

P.S. Of course, if anyone has a pair of Wild 10x high eyepoint oculars for
sale, that would also solve my problem (I'd also consider other
magnifications). (I'm also looking for accessory objective lenses and
various other accessories. Please contact me off-list if you've got anything
for sale or trade.)








From: Dr. Gary Gaugler :      gary-at-gaugler.com
Date: Fri, 04 Feb 2000 15:02:28 -0600
Subject: Re: Help:A comparison of optics

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At 09:20 AM 2/4/00 , you wrote:
} } A client of ours wonders about a comparison between Zeiss and Olympus
} } optics. The Zeiss 100X lense cost $10,000, and the Olympus about $4,000:
} } does that mean the Olympus is inferior?  That's what our client's
} } colleagues imply. What measure of lense function is relevant? Does Olympus
} } (or Zeiss) have a specification, like 'resolves particles to 0.1 micron'
} } or something like that?  Is there a measure of distortion? Best
} } regards, Steve NiemelaVayTek, Inc.305 West Lowe AvenueFairfield, IA
} } 52556email: sniemela-at-vaytek.com Phone: 515.472.2227Fax: 515.472.8131web:
} } www.vaytek.comgeneral email: vaytek-at-vaytek.com
} }

The NA is very important in achieving high resolution. The other factor is
the NA of the condenser. An Abbe condenser is easily out gunned by
the aplanatic condenser (NA=1.4). Olympus makes two PlanAPO 100X oil
objectives. One has an NA of 1.3 while the other is 1.4. The 1.4 lens
costs today new about $6000. The lower NA lens is about $4500. So this
is probably the one you are comparing. I had a Zeiss PlanAPO and recall
that it had an NA of 1.3. It was terrible. I replaced it with the Olympus 1.4
and got major improvements. Of the Zeiss objectives, their 63X PlanAPO
is probably the best one they have made. The rest are so so. Some are
easily beaten by Olympus flourites. Zeiss is more expensive because it
is Zeiss, not necessarily because it is better. The cost of manufacturing
in Germany is very high compared to Japan and other places that
Olympus uses. The emerging problem I see is that Olympus is moving much
of its scope production to China. I have purchased all of the Olympus
items I need and that were made in Japan before being stuck with Chinese
Olympus. It may reach a point where Zeiss is a better product despite the
higher cost.

BTW, the 1.4NA lens at $6000 reflects the recent 10% across the board
price increase by Olympus.

gary g.





From: Sinkler, Wharton :      wharton.sinkler-at-anlw.anl.gov
Date: Fri, 04 Feb 2000 14:14:10 -0700
Subject: Re: TEM:finding the parallel beam

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Listers,

There are important differences between older and more recent TEMs, which go
to the heart of this discussion.

On older dedicated HREM's (LaB6) imaging is done with the beam at or close
to crossover on the specimen, and beam convergence is determined by the user
using the C2 aperture. On newer TEM's such as the JEOL 2010 (FEG or not),
you need to set C2 so crossover is pretty far from the specimen plane in
order to get decent coherence for imaging. Whenever this is the case, the
C2 aperture isn't incoherently filled, and some of the older theory won't
apply (regardless of whether the machine is or isn't a FEG).

For example, the standard method of determining convergence in an image is
by switching to diffraction under the same conditions used to image, and
measuring the disc diameter. (see e.g. O'Keefe and Sanders, Acta Cryst A31,
p.307). This assumes the beam being near or at crossover on the specimen
aperture. It won't work if the beam is far from crossover on the specimen,
because now the incident angle depends on position on the specimen. The
disc diameters in diffraction only indicate what the total range of incident
angles is over the entire illuminated area. But the area visible in the
HREM image is much smaller than this, and over this latter area the range of
illuminating angles is smaller. This is just another way of saying that the
C2 aperture isn't incoherently filled. If it were, would be impossible to
form a shadow image of the C2 aperture at the specimen plane, as one does by
spreading the beam.

Think of the illumination on the specimen as a convolution of the source
image with the circular C2 aperture: When the beam is at crossover, the
aperture part is approximating a delta function - we see the source in the
image. When the beam is spread very far, the illumination is a nice shadow
image of the C2 aperture, with a little blurriness at the edge because of
convolution with a source which is not quite point-like.

In the latter case, the angular spread locally is given by the source size
(demagnification) in the diffraction pattern. This can be imaged by
switching to diffraction and focusing with the intermediate lens to obtain
sharp source images.

This doesn't depend on whether the machine is FEG or not, though the source
will be very much smaller if it is. I have posted an example in which a
tungsten filament is imaged sharply by defocusing with the intermediate lens
with the illumination just slightly defocused. This should be a square
pattern, so note that the optics introduce significant distortions in this
extreme case.

http://www.numis.nwu.edu/internet/Staff/wharton/sources.jpg


++++++++++++++++++++++++++++++++++++++++++++++++++
Wharton Sinkler
Argonne National Laboratory West
P. O. Box 2528
Idaho Falls, ID 83403

}
}
} On Thu, 3 Feb 2000, Thomas, Larry wrote:
}
} } Laurie,
} }
} } Now I'm really confused. I used to think I understood this somewhat.
} }
} } 1. In the JEOL 2010F FEG (the "all bets are off" case in your item#7),
} } changing the illumination spread by changing C2 profoundly moves the
} } "diffraction plane" relative to the objective aperture. [There are actually
} } two
} } objective apertures at different heights in this microscope, but that's
} } beside
} } the point]. By diffraction plane, I mean the plane below the objective lens
} } where the diffraction spots are in sharp focus. In other words (assuming
} } fixed
} } OL and condenser minilens excitations), in this microscope there is only one
} } C2
} } value that allows the diffraction spot pattern and OL aperture to be in focus
} } simultaneously. If you focus on the OL aperture (with the intermediate
} } lens),
} } the spot pattern can be focused at only this one C2 setting. At any other C2
} } setting, you can focus the diffraction pattern with the intermediate lens
} } ("diffraction focus") but the OL aperture will then be defocused. Changing
} } the
} } condenser minilens excitation changes the diffraction plane (relative to the
} } OL
} } aperture) to a different C2 setting. This is actually quite a useful feature
} } of
} } the microscope, e.g., for getting very intense focused diffraction patterns,
} } but
} } a different story.
} }
} OK, the bets off case. In a FEG you have a somewhat coherent
} range of incident directions. The "diffraction pattern" is therefore
} something which is effected by the coherent aberrations of the microscope,
} unlike the case with incoherent illumination. Consider the case with
} focussed illumination, when the diffraction pattern shows an image of
} the condensor aperture. In a FEG As a you can "focus" this (coherent)
} image to a small spot by going out of focus in a true sense for the
} diffraction pattern. This gives you a spot-like pattern, but you will also
} see severe distortions at higher angles. With incoherent illumination you
} cannot do this and going out of focus (in the diffraction pattern) will
} give in general a blurred image of the condensor aperture - life is
} simple.
} What you need to do is focus "real diffraction" features, e.g.
} Kikuchi lines, then not play with the diffraction focus at all. As you
} change C2 the size of the spots will grow or shrink, this is fine.
}
} } 2. The reason for choosing to focus the diffraction spots rather than
} } the K-lines is --to put it simply-- that's where the intensity is. In
} } amplitude
} } contrast imaging (conventional brightfield and darkfield imaging), the
} } intensity
} } contribution to the image is limited by the OL aperture. It's really
} } important
} } to have the defining aperture and diffraction spot pattern in focus
} } simultaneously. In addition, if I aim to measure diffraction spot patterns
} } there is little incentive to focus the K-lines instead of the spots.
} }
} Yes, you can always get a "spot" pattern with a FEG, and it may
} be prettier. However, beware the distortions which make doing
} measurements from it very dangerous.
}
} } 3. Textbooks such as Hirsch et al. often discuss the positional error
} } in SA diffraction due to Cs. What readers often miss is the consequence of
} } the
} } fact that the error depends strongly on the diffraction angle. If
} } measurements
} } are confined to the low-index (low-angle) reflections, the selecting area can
} } be
} } reduced (to 200 nm or so at 200 kV) without big positional errors. Maybe
} } that's
} } an example of differences in experimental and theoretical viewpoints.
} }
} Hirsch et al is just a good reference to read, about this as well
} as many other things!
}
} } 4. Reducing the condensor aperture size will give less illumination,
} } but it won't give parallel illumination. It will change the size of the
} } diffraction spots, but not their focus.
} }
} It makes it closer to parallel. Of course, with really parallel
} illumination there is no intensity!
}
} } 5. I'm not quite sure why the illumination spread has such a strong
} } effect on the apparent position of the diffraction plane relative to the OL
} } aperture, but suspect it arises from the action of the strong
} } condensor-objective in this microscope. I would also question that it has
} } much
} } to do with coherent/incoherent beams in a FEG or with prefield/postfield Cs
} } effects.
} }
} Sorry, I don't think that is correct. I would suggested that
} people look at the distortions with a standard sample and see how bad they
} can be.
}
} } Larry
} }
} }
} } Larry Thomas
} } Pacific Northwest National Laboratory
} } MSIN P8-16
} } P.O. Box 999
} } Richland, WA 99352
} } Phone: (509)372-0793 Fax: (509)376-6308
} } Email: mailto: Larry.Thomas-at-pnl.gov
} }
} }
} }
} } ----------
} } From: L. D. Marks
} } Reply To: L-marks-at-nwu.edu
} } Sent: Thursday, February 3, 2000 8:06 AM
} } To: Radostin Danev
} } Cc: MSA listserver
} } Subject: Re: TEM:finding the parallel beam
} }
} } ------------------------------------------------------------------------
} } 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
} } -----------------------------------------------------------------------.
} }
} }
} } Sorry, but I think we are getting a little confused here.
} }
} } 1) The diffraction plane is (by definition) the objective lens
} } focal length below the sample - it is the focus of parallel
} } directions coming from the sample. The focal length (and
} } parameters such as Cs, Cc) change rapidly with the objective lens
} } current so an eucentric position makes life simple but is not
} } necessarily the best condition to use. (For HREM you generaly
} } want to have the objective lens stronger and reduce Cs.)
} }
} } 2) In general C2 does not change the focal length at all. You
} } can have some effect from coupling of the magnetic fields, but
} } all C2 is supposed to do is change the convergence angle.
} }
} } 3) The objective aperature is (approximately) in the back-focal
} } plane, given that it is finite and height adjustments are only
} } done coarsely (for a specific value of the objective current
} } only).
} }
} } 4) The selected area aperature is (approximately) in the same
} } plane as the object for the same reasons as above. Remember that
} } there are positional errors in SA diffraction due to Cs (see
} } Hirsch et al for instance).
} }
} } 5) You should focus Kikuchi-lines, not go for the smallest spot,
} } since these are real object in the diffraction pattern. What you
} } will see is then representative of you incident beam, sometimes
} } a Gaussian range of directions.
} }
} } 6) The simplest way to get more parallel illumination in general is
} } to reduce the condensor aperture size.
} }
} } 7) All bets are off if you have a FEG. Rather than having incoherent
} } illumination things like the Cs of the prefield (above the sample)
} } and postfield (below) add coherently making it much more complicated.
} } For instance, the illumination angle varies across the field of view.
} } While this is also there with LaB6, it is a weak effect (except for
} } certain classes of diffracted beams, e.g. those with a screw-axis).
} } With
} } a FEG you can get a "spot" pattern many ways, and most of these will
} } have
} } large aberrations (e.g. pincushion).
} }
} } ++++++++++++++++++++++++++++++++++++++++++++++++
} } Laurence Marks
} } Department of Materials Science and Engineering
} } Northwestern University
} } fax: (847) 491-7820
} } mailto:l-marks-at-nwu.edu
} } http://www.numis.nwu.edu
} } ++++++++++++++++++++++++++++++++++++++++++++++++
} }
} }
} }
}
} ++++++++++++++++++++++++++++++++++++++++++++++++
} Laurence Marks
} Department of Materials Science and Engineering
} Northwestern University
} fax: (847) 491-7820
} mailto:L-marks-at-nwu.edu
} http://www.numis.nwu.edu
} ++++++++++++++++++++++++++++++++++++++++++++++++
}
}





From: Jaci Lett :      jmlett-at-cid.wustl.edu
Date: Fri, 4 Feb 2000 16:01:45 -0600
Subject: TEM: B-glucuronidase localization

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We are looking to localize B-glucuronidase in the mouse cochlea. We'd like
to stain for the enzyme and still be able to decalcify the cochleas and
process them into Epon-Araldite for LM and TEM without losing the reaction
product. Does anyone have any suggestions for staining or localization
protocols for this enzyme? I'm not certain whether the tissue will be taken
all the way to TEM, but I do know we'll be looking at LM (1 to 4 microns).
I found a method by Hayashi and Hayashi ("Electron Microscopic Cytochemistry
and Immunocytochemistry in Biomedicine", CRC Press, 1993) using hexazonium
pararosanilin (red in LM and electron dense for EM) on 40 micron non-frozen
sections (and then processed for EM) that sounds appropriate, but I'm
concerned that it may not work en bloc (the cochlea is a twisty, windy
beasty).

I posted this request a few months ago and received no assistance (just
individuals also interested in similar protocols and a company promoting
their antibody).

Thanks so much for any direction I can get,

Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.edu

Fay and Carl Simons Center for Biology of Hearing and Deafness
Central Institute for the Deaf
818 S. Euclid Ave.
St. Louis, MO 63110

voice: 314-977-0257 fax: 314-977-0030





From: Jonathan Barnard :      Jonathan.Barnard-at-Angstrom.UU.SE
Date: Sat, 05 Feb 2000 00:45:21 +0100
Subject: TEM: Jeol 2010 question

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So the problem as I have understood it is:

===} changing C3 (brightness) leads to a shift of the bright field (BF)
beam in the back focal plane (BFP) causing the objective aperture to appear
off centre after converging the beam?

If you have aligned the gun properly then I suspect that the (shift) wobble
alignment has not been done properly.
When this button is pressed the microscope goes over to diffraction and you
should see a pair of discs or spots (which represent the two extremes in
tilt when the beam is shifted to its own extremes). The beam tilts are
adjusted to bring the two discs/spots into coincidence. This is done for
both the X and Y shift coils separately.
The X wobble button shifts the beam back and forth (X shift coils), but in
the back focal plane the spot or disc should remain stationary. This is
what you are trying to achieve with this alignment procedure. Afterwards
this should give you a beam that expands about the same point in the back
focal plane. You should find that the objective aperture remains centred in
both diff and image modes after doing this alignment.

As for the crossovers, yes you are right, they do move up and down when
changing the brightness. The back focal plane remains stationary (by
definition as L.Marks said). Anything other than a parallel beam should
give you a disc in the BFP (which represents the angular distribution of
rays incident on the objective lens).

What did you mean by a voltage centre? My interpretation of this is that
the source high tension is wobbled and any shift in the (previously
focussed) image is minimised by tuning the beam tilts. This alignment would
be used for energy filtered TEM (EFTEM) when doing elemental mapping on a
Gatan Imaging Filter (GIF) for example.



********************************************************
Dr Jonathan Barnard

Analytical Materials Physics
The Angstrom Laboratory, Uppsala University
P O Box 534, SE-751 21 Uppsala, Sweden
Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
http://www.angstrom.uu.se/analytical/home.html

********************************************************





From: Stephen A. Shaffer :      sshaffer-at-microdataware.com
Date: Fri, 04 Feb 2000 15:47:58 -0800
Subject: LM Meeting Announcement, San Francisco Microscopical Society

Contents Retrieved from Microscopy Listserver Archives
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The San Francisco Microscopical Society Announces its next regular
meeting:

Wednesday, February 9, 2000

Randall Museum
199 Museum Way
San Francisco, CA 94114-1429
(415) 554-9600

7:00 pm

Topic: Introduction to Phase Contrast Microscopy
by Robert D. Griffin, President, SFMS
And: Annual Business Meeting and Election of Officers

Further information, including an important Letter to the Members from
President Griffin, is available at http://www.microdataware.com/sfms .

The San Francisco Microscopical Society (SFMS) is a small, informal
group of light microscopists who meet on a monthly basis to discuss
topics of common interest. All are invited and welcome, regardless of
knowledge level or professional field.





From: James Pawley :      jbpawley-at-facstaff.wisc.edu
Date: Sat, 5 Feb 2000 14:39:24 +1100
Subject: 5th Live-cell Course at UBC: June 19-29

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Dear Steve,
Because of the nomenclature and the way manufacturers determine their
specifications, it is really difficult to compare optics on paper. When I
find myself in these types of discussions, I find it more useful to compare
optics of similar list price rather than specifications because generally
speaking, in our industry, the more things cost, the more care is taken in
their manufacture. Olympus has a new 100X objective with an NA of 1.65 that
sells for $10k. That would be the optic I would compare to the $10k Zeiss.

Shane Collins
Scientific Instruments

-----Original Message-----
} From: Steve Niemela [mailto:sniemela-at-vaytek.com]
Sent: Thursday, February 03, 2000 2:05 PM
To: microscopy-at-sparc5.microscopy.com


To: {Microscopy-at-MSA.Microscopy.Com}


SECOND ANNOUNCEMENT: UBC LIVE-CELL COURSE: Please register by March!!

Hello all,

The faculty for the 2000 UBC 3D Live-cell Microscopy Course is now set and
we welcome several new faces, including Stephan Hell, Andreas Kriete and
Steve Potter. The whole list is below.

o Stephen Adams University of California-SD
o Ping Chin Cheng SUNY, Buffalo
o Elaine Humphrey Univ. of British Columbia
o Stephan Hell Max Planck, Goettingen
o Ted Inou Universal Imaging, PA
o Iain Johnson Molecular Probes, OR
o Larry Keenan Cell Robotics, NM
o Ernst Keller Carl Zeiss, Thornwood, NY
o Andreas Kriete University of Geissen
o Felix Margadant University of Sydney
o Tim Murphy University of British Columbia
o Jim Pawley University of Wisconsin-Madison
o Steve Potter California Institute of Tech
o Jason Swedlow University of Aberdeen
o Michael Weis Agriculture Canada


Basic info is below but you can get the entire brochure, including links to
faculty home pages, at

http://www.cs.ubc.ca/spider/ladic/course/brochure.htm

Cheers,

Jim P.

Fifth Annual INTERNATIONAL 10-Day Short Course on

3D Microscopy of Living Cells
June 19 - 29, 2000



Fourth, Post-course Workshop on

3D Image Processing,
July 1 - 3, 2000



Organized by Prof. James Pawley,
(University of Wisconsin-Madison)
(SEE SABBATICAL ADDRESS AT END OF MESSAGE)


in association with Dr. Elaine Humphrey
UBC BioSciences Microscopy Facility:
University of British Columbia
Vancouver, BC, Canada



DATES

Applications must be received by March 1, 2000
Deposit due April 15, 2000
Registration 5:00 - 7:00 PM Sunday, June 18, 2000
First Lecture 7:30 PM Sunday, June 18, 2000
Live-cell Course ends, noon Thursday, June 29, 2000


APPLICATIONS DUE BY MARCH 1, 2000


More information at :
http://www.cs.ubc.ca/spider/ladic/course/bulletin.html

or

REGARDING COURSE ORGANIZATION:


Prof. James B. Pawley, (on Sabbatical)
Room LG 10, Madsen Building, F-09,
University of Sydney, NSW, 2006
Australia

Ph. 61-2-9351-7548/2351
FAX 61-2-9351-7682
JBPAWLEY-at-FACSTAFF.WISC.EDU

REGARDING APPLICATIONS

Dr. Elaine Humphrey,
Biosciences EM Facility
Biosciences Building
Univ. of British Columbia
6270 University Blvd.
Vancouver, BC, V6T-1Z4




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, an intensive
eleven-day residential course concentrating on all aspects of 3D Microscopy
of Living Cells will be held at the University of British Columbia, in June
of 2000. The course includes 4 days on 2D techniques, 6 days of 3D
techniques and a summary presentation. It covers basic microscopy to the
highest level confocal microscopy. (A half-day Pre-course is offered for
any who may need to brush up on basic optics!).

Topics include:
o Quantitative 2D light microscopy
o How to keep your cells alive
o 3D imaging in confocal microscopy
o Widefield/deconvolution techniques
o Two-photon excitation microscopy
o Fluorescent and backscattered light signals
o Dye design, characteristics and use
o Pixelation: The Nyquist Criterion
o Lasers and laser tweezers
o Objectives and aberrations
o Scanning-systems: AODs and mirrors
o Optimal pinhole size/photon efficiency
o Detectors: operation and performance
o Video-rate confocal imaging
o Measuring ion concentrations
o Display and measurement of 3D data

Morning lecture/demonstrations lead to hands-on laboratory exercises each
afternoon that will utilize most of the commercial instruments currently
available for 3D microscopic imaging. Students will work in groups of 3 or
4 throughout the discussion and laboratory sessions, and may complete a
live-cell 3D study on their own specimens. In the first four years, over
100 students from 22 countries have attended. Last year, 11 separate, 3D
microscopical workstations were available for student use under the
supervision of a 17-member international faculty. We expect to have even
more workstations in 2000. Including manufacturers representatives, the
teacher/ student ratio will be almost 2:1.

INTERNATIONAL FACULTY

o Stephen Adams University of California-SD
o Ping Chin Cheng SUNY, Buffalo
o Elaine Humphrey Univ. of British Columbia
o Stephan Hell Max Planck, Goettingen
o Ted Inou Universal Imaging, PA
o Iain Johnson Molecular Probes, OR
o Larry Keenan Cell Robotics, NM
o Ernst Keller Carl Zeiss, Thornwood, NY
o Andreas Kriete University of Geissen
o Felix Margadant University of Sydney
o Tim Murphy University of British Columbia
o Jim Pawley University of Wisconsin-Madison
o Steve Potter California Institute of Tech
o Jason Swedlow University of Aberdeen
o Michael Weis Agriculture Canada

TUITION

Course tuition is $2,150 US and includes lunches. On receipt of 50%
deposit, students will receive preliminary group assignments and the
textbook, Handbook of Biological Confocal Microscopy, (Plenum, 1995). The
tuition fee includes the textbook, a binder of handouts, and tickets for
the Opening Reception, the Manufacturer's Reception and the Beach Party.
Accommodations and meals other than lunch are not included in the tuition
fee. The Pre-course is $100 US.

APPLICATIONS

Applicants will complete a questionnaire to assess knowledge level and
field of interest. Enrollment is limited to about 24 participants.
Selection will be made on the basis of background and perceived need.
Those without previous LM experience will be provided with basic texts on
request to read before the course begins and are encouraged to take the
Pre-course on the afternoon of June 18.


Application forms, and other course information from this and past years,
can be downloaded from the WWW site at:

http://www.cs.ubc.ca/spider/ladic/course/bulletin.html

or obtained from:

Dr. Elaine Humphrey,
Biosciences EM Facility
Biosciences Building
Univ. of British Columbia
6270 University Blvd.
Vancouver, BC, V6T-1Z4
Phone: 1-604.822-3354
FAX: 1-604.265-5315
Email: ech-at-unixg.ubc.ca.


Application deadlines:

Application forms are due by March 1, 2000!
Deadlines are early to facilitate setting up groups. Successful applicants
will be notified by April 1, and a deposit of 50% must be received by April
15, 2000 to reserve your position. In general, deposit refunds are only be
possible if your position can be filled from the waiting list. The
remainder of the fees is due at registration.

DATES

Applications must be received by March 1, 2000
Deposit due April 15 2000
Registration 5:00 - 7:00 PM Sunday, June 18, 2000
First Lecture 7:30 PM Sunday, June 18, 2000
Live-cell Course ends, noon Thursday, June 29, 2000


TEACHING PHILOSOPHY

As befits teaching in an area at the boundary of "what is now known,"
lecturers have been chosen based on their expertise as scientists working
in the field rather than because they all agree. They are encouraged more
to be provocative than to be prosaic. Students should expect discussion in
areas where differences of opinion exist.

Prior to the course, students will be organized into groups and encouraged
to communicate by email/phone, about the "Living-cell" group projects that
they will pursue during the course and that will be presented to the class
on the last day of the course. It has been found that group interactions
make best use of students' prior experience and can be very effective in
teaching the practical skills covered in a hands-on course of this type.

ARRANGEMENTS FOR LIVE SAMPLES

Students must contact Dr. Elaine Humphrey to make necessary arrangements
for the transport and maintenance of cell lines etc. needed for their
projects. Organisms linked in any way with human disease are not permitted
because of safety considerations. Transport and customs arrangements for
living specimens are entirely the responsibility of the student.
Students also attending the 3D Image Processing Course, may be able to
analyze, process and display some of the 3D data collected from their
specimens


********************************************************************************

Fourth Annual

3D MAGE PROCESSING WORKSHOP

The workshop will cover 3D image processing for measurement and display.
Enrollment is limited to those attending the 3D Microscopy course.
Tuition : $850 US (lunches and snacks incl.)

WHO SHOULD ATTEND?

The course is designed for biologists working with multidimensional and
possibly multicolor microscopical data sets. Getting the data is only half
of the battle. Image data in 3, 4 or even 5 dimensions may be difficult to
store let alone analyze or display. This course is to help students
understand the hardware and software aspects of this problem and give them
the techniques they need to make the best use of their data.

The course is designed for biologists who need to make measurements on 3D
microscopical data sets and then display the results in an effective
manner. The course will be taught on SGI, Macintosh and IBM-compatible
computers. A wide variety of software designed for the 3D microscopy market
will be described, demonstrated and available for use.

Workshop Organizers

o Andreas Kriete Giessen University, DE
o Felix Margadant University of Sydney, Au

Faculty

o Ping Chin Cheng State U. of New York, Buffalo
o Chris MacLean Vaytek Inc., IA

PLAN OF INSTRUCTION
Classes will meet from 8:30-12:00 and 1:00-6:00 with lecture-demonstrations
followed immediately by hands-on laboratory sessions using a variety of
workstations. Students will "learn-by-doing" with two to a machine. Lab
handouts will describe some specific exercises to be performed on "canned"
data sets. Facilities and supervision will be available until 11:00 PM, for
students to work on their own data.
****************************************
Prof. James B. Pawley, (on Sabbatical) Ph. 61-2-9351-7548/2351
Room LG 10, Madsen Building, F-09, FAX 61-2-9351-7682
University of Sydney, NSW, 2006 Australia JBPAWLEY-at-FACSTAFF.WISC.EDU
Come to the 5th 3D Live-cell Microscopy Course, June 19-29, 2000, UBC, Canada.
Info and forms at: http://www.cs.ubc.ca/spider/ladic/course/bulletin.html
"A scientist is not one who can answer questions but one who can question
answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39




From: Stan Rice :      srice-at-alpha.utampa.edu
Date: Fri, 4 Feb 2000 21:55:32 -0600
Subject: LKB Nova ultramicrotome

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I recently inherited a used LKB Nova ultramicrotome from a medical
facility in town. The previous owner could not locate the operating
instructions and I am at a loss to figure out its proper operation. Does

anyone on the list have instructions for operation of this instrument??
I would be willing to reimburse anyone for a copy of the instruction
manual.

Dr. Stanley A. Rice
University of Tampa
srice-at-alpha.utampa.edu
(813) 253-3333 Ext. 3340

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From: Patricia A. Glazebrook :      PGlazebr-at-Research.metrohealth.org
Date: Fri, 4 Feb 2000 21:57:44 -0600
Subject: Cy5 filtercube today on a upright flourescent microscope

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I tried a Cy5 filtercube today on a upright flourescent microscope equiped
with a Diagnostics Instrument Spot Camera. My tissue had been
immunolabelled with Cy5 and I did not expect to see thru the microscope. So
we took pictures with the Spot Camera and the immunolabeling seemed to
work. The problem was that there was uneven illumination in the pictures,
half moon of brightness and the rest of the field dark. The salesperson did
check that the mercury bulb was aligned correctly. Also there was no
ambient light from the room causing the shadow.

Anyone with an idea  of what is causing this problem?

Thank-you, Pat Glazebrook







From: Lucille A. Giannuzzi :      lag-at-mail.ucf.edu
Date: Fri, 4 Feb 2000 21:59:21 -0600
Subject: TEM Specimen Prep short course

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TEM Specimen Preparation Short Course!!!!

With emphasis on Tripod Polishing and Focused Ion Beam Specimen Preparation
....

..will be offered in conjunction with the Joint Annual Meeting of the
Florida Society for Microscopy and the Florida American Vacuum Society in
Orlando at the University of Central Florida March 15-17, 2000.

Instructors:

Ron Anderson, IBM
Fred Stevie, Lucent Technologies
Lucille Giannuzzi, UCF

Vendor Sponsors:

FEI Company
Micro Optics
South Bay Technology

For registration material please contact Lucille Giannuzzi at lag-at-mail.ucf.edu

*******************************************************************
Lucille A. Giannuzzi, Ph.D.

Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng.,
University of Central Florida,
PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA
phone (407) 823-5770 fax (407) 823-0208 email lag-at-mail.ucf.edu

Director, UCF/Cirent Materials Characterization Facility, 12443 Research
Parkway, Suite 305
Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321
--------------------------------------------------------------------
"Good judgement comes from experience.

Experience comes from making bad judgement."

Mark Twain
********************************************************************






From: Roger Moretz :      rcmoretz-at-excite.com
Date: Fri, 4 Feb 2000 20:52:16 -0800 (PST)
Subject: Re: Help:A comparison of optics

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The responses to this thread have been outstanding. Yet from my perspective
the unasked and thus unanswerable question is: what is the purpose for the
lens? A couple of the responses have brushed the issue, but it still hasn't
been specifically stated. Assuming that resolution is the main issue, the
concerns of flatness of field, NA, color correction, working distance, etc,
all have to be factored into the evaluation of the different lenses. It
should be possible to demo both (or any of a set of) lenses under the lab
and experimental conditions of real life. Only then can one truly determine
which lens is the "best" (or more correctly, the appropriate) lens. All of
the theory and specifications in the world don't matter if the lens doesn't
aid the microscopist in obtaining the data necessary to carry out the work.
One thing that is unalterable is that even if one is considering a number of
"infinity-corrected" lenses from different manufacturers, they may not be
truly interchangeable. Make sure the lens works on the microscope body.
Then make sure it provides the performance needed. That's the right lens.

Roger Moretz


On Thu, 3 Feb 2000 16:04:55 -0600, Steve Niemela wrote:

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} To: {Microscopy-at-MSA.Microscopy.Com}
} Subject: A comparison of optics
} Date: Thu, 3 Feb 2000 10:57:19 -0600
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} X-Mailer: Microsoft Outlook Express 5.00.2615.200
} X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2615.200
}
} A client of ours wonders about a comparison between Zeiss and
Olympus
} optics. The Zeiss 100X lense cost $10,000, and the Olympus about
$4,000:
} does that mean the Olympus is inferior?  That's what our client's
} colleagues imply. What measure of lense function is relevant? Does
Olympus
} (or Zeiss) have a specification, like 'resolves particles to 0.1 micron'
} or something like that?  Is there a measure of distortion? Best
} regards, Steve NiemelaVayTek, Inc.305 West Lowe AvenueFairfield, IA
} 52556email: sniemela-at-vaytek.com Phone: 515.472.2227Fax: 515.472.8131web:
} www.vaytek.comgeneral email: vaytek-at-vaytek.com
}
}
}


Roger C Moretz, Ph.D.





_______________________________________________________
Get 100% FREE Internet Access powered by Excite
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From: Ken Tiekotter :      tiekotte-at-up.edu
Date: Sat, 5 Feb 2000 00:36:00 -0800 (PST)
Subject: Re: Aqueous mounting medium

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Lesley,
How about CMC (Refractive Index=1.37). CMC10 (R.I.=1.39), CMCP-9
(R.I.=1.40) and CMCP-10 (R.I. 1.41)? This is a mixture of Polyvinyl
alcohol (5%), phenol(25%), lactic acid (25%), and water (45%).

I believe you can get this from Polysciences Inc. This mountant causes
problems with most stained specimens, but may work for you. Is glycerine
jelly auto-fluorescening?

-Ken

--------------
Ken Tiekotter
Dept. of Biology
The University of Portland
5000N. Willamette Blvd.
Portland, OR 97303







From: Ken Tiekotter :      tiekotte-at-up.edu
Date: Sat, 5 Feb 2000 00:45:21 -0800 (PST)
Subject: Re: Help:A comparison of optics

Contents Retrieved from Microscopy Listserver Archives
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Dan,
Mixing and matching objectives is one thing, while doing the same for
oculars is probably not visually as big an issue. However, in highly
corrected systems, compensating eyepieces are a critical consideration if
one wishes to maintain image quality, i.e., plan apo objectives require
compensating oculars to maintain the correction of the apochromatic
objective. This is especially true for color reproduction in color
photomicrograph. This is not as important in black and white
photomicrography.

-Ken

----------------
Ken Tiekotter
Dept. of Biology
The University of Portland
5000 N Willamette Blvd.
Portland, OR 97303


On Fri, 4 Feb 2000, Dan Freidus wrote:

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} -----------------------------------------------------------------------.
}
}
} -----Original Message-----
} } From: Ken Tiekotter {tiekotte-at-up.edu}
} ..One does not mixed and match objectives from different systems merely for
} the convenience of pricing....
} ----------------------------------------------------------------------------
} ---------------------------------------------------------------------------
}
} Of course, I've heard this sort of statement many times before. But has
} anyone ever done any comparisons of mixed systems to see if there are
} certain brands that are particularly incompatible or particularly
} compatible?
}
} I ask this both as a general question and because I use a Wild M7 for which
} I need a pair of high eyepoint oculars. I don't need the adjusting collar
} built into current models and haven't been able to find an older pair used.
} But before I buy a Nikon/Olympus/ etc. pair I'd love to know how different
} the complete system will be from what I've got now.
}
} I know many people who have used third party photo eyepieces, in just the
} situation where you'd think quality would be of the utmost concern. Dealers
} are often not the best source of info since they tend to represent one brand
} and, even when they are completely honest, rarely have experienc ewith a
} wide range of brands (especially mixed up.
}
} Dan Freidus
} freidus-at-wwnet.com
}
} P.S. Of course, if anyone has a pair of Wild 10x high eyepoint oculars for
} sale, that would also solve my problem (I'd also consider other
} magnifications). (I'm also looking for accessory objective lenses and
} various other accessories. Please contact me off-list if you've got anything
} for sale or trade.)
}
}
}
}
}
}





From: Paul Webster :      pwebster-at-hei.org
Date: 05 Feb 00 12:50:25 -0800
Subject: RE: Aqueous mounting medium

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Lesley Weston {lesley-at-interchange.ubc.ca}
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From: Paul Webster :      pwebster-at-hei.org
Date: 05 Feb 00 12:50:25 -0800
Subject: RE: Aqueous mounting medium

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Reply to: RE: Aqueous mounting medium
Dear Lesley,

You can make a very useful mounting medium from inexpensive ingredients.
The main ingredient is poly(vinyl) alcohol which is soluble in water but will dry without shrinking too much. The water solubility means you can take previously mounted slides, rehydrate them and use them again.

Fading of fluorescent dyes can be retarded by including anti-fade compounds.

You can find the recipe at {http://www.hei.org/htm/moviol.htm} .

Details of anti-fade compounds can be found here (at the bottom of the page) {http://www.hei.org/htm/ifse.htm} .
Ths site is getting a little outdated so if you feel there are details missing then please contact me with your questions.

Many thanks to Heinz Schwarz of Tuebingen for writing these protocols.

Regards,

Paul Webster

Paul Webster, Ph.D.
Associate Scientist & Director
Ahmanson Advanced Electron Microscopy & Imaging Center
House Ear Institute
2100 West Third St.
Los Angeles, CA 90057

Phone: (213) 273-8026
Fax: (213) 413-6739
e-mail: pwebster-at-hei.org
http://www.hei.org/htm/aemi.htm





From: donald j marshall :      dmrelion-at-world.std.com
Date: Sun, 6 Feb 2000 07:21:55 -0500 (EST)
Subject: solid state emitters

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I would call this a typical PARTIAL judgment.
What you see is what you get. Thee only way, to the best of my experience,
is to get both objectives, side by side if possible, and to do your own
evaluation. I have honestly doubts concerning those values set on
objectives.
Things change and optic too and sometimes faster that we could expect.
Norm
----- Original Message -----
} From: Dr. Gary Gaugler {gary-at-gaugler.com}
To: MSA listserver {Microscopy-at-sparc5.Microscopy.Com} ; {sniemela-at-vaytek.com}
Sent: Friday, February 04, 2000 5:02 PM


I am interested to know what the present state of the art is re: solid state
electon emitters, total electron beam currents available and also beam
current densities. If anyone could point out a recent review article, I
would appreciate it. Thanks,

Don Marshall

Donald J. Marshall
Relion Industries
P.O. Box 12
Bedford, MA 01730
Ph: 781-275-4695
FAX: 781-271-0252
email dmrelion-at-world.std.com

Cathodoluminescence, mass spectroscopy, electron beam technology


"A weed is a flower out of place."

(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)




From: Bill Miller :      microbill-at-mohawk.net
Date: Sun, 06 Feb 2000 08:43:16 -0500
Subject: Re: Help:A comparison of optics

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In fact, if you're really interested in getting the BEST lens, try at
least three examples of the ones you are interested in . All high NA lenses
must meet some minimal standard but some are significantly better than that
minimum and the only way to find one that is is to try THE lens you are
going to buy - not just an example.

Bill Miller

At 10:04 PM 2/5/00 -0400, Norm wrote:
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From: VALLARAY-at-aol.com
Date: Sun, 6 Feb 2000 16:10:45 EST
Subject: Re: Electron diffraction simulations

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Erik,

Do not have difraction sofrware, but here are some URLs where you can
download free simulation software I use for e-beam to solid interactions,
hope it may be useful for you.

Casino is a freeware for monte carlo simulations:
http://www.gme.usherb.ca/casino/

Monte carlo resources:
http://dsa.dimes.tudelft.nl/pattern_definition/monte/monte.html

My FAVORITE monte carlo software:
http://web.utk.edu/~srcutk/htm/simulati.htm

Valery Ray, MSEE,
SEM engineer
Applied Materials
001-208-8413744




From: Richard Hey :      richardh-at-jeoleuro.com
Date: Sun, 6 Feb 2000 18:32:21 -0600
Subject: Re: Parallel beam

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Hi everybody
While this has been an interesting discussion we seem to have got away from
the original question.

This is the method I use - after of course ensuring normal alignments and
adjustments have been carried out

Insert the objective aperture with a specimen in the beam. Focus the image
of this aperture in Selected Area Diffraction using the first intermediate
lens focus.

Focus the diffraction pattern (i.e. the smallest zero order spot) with the
final condenser lens, this will give a parallel beam incident upon the
specimen.

To test this, remove the objective aperture and insert the largest selected
area aperture. If the beam is parallel moving this aperture will not move
the diffraction spot.


------------------------------------------------------------

Tilt and Shift alignment wobblers in the 2010

Tilt - The purpose of this alignment is to ensure that when the beam is
tilted it doesn't move on the specimen. This is double tilt correction and
depends on using double deflector coils.

Shift - Double shift correction is meant to ensure that when the beam is
shifted it doesn't tilt. The operation of this alignment is dependent upon
the value of the first intermediate lens. The best value for this alignment
is a parallel beam condition as described above. By the very nature of the
beast it will be found to be impossible to perform this alignment correctly
at some CBD conditions. This is meant to be a convergent beam mode after all
not a parallel beam mode.

I understand that this is also referred to as tilt and shift purity.

Richard Hey

The views expressed herein are purely my own and do not reflect those of my
employer

Richard Hey
Technical Support Engineer
Jeol (UK) Ltd
Phone: (44) 1707 377117






From: Qiaoling Jin :      jinq-at-pilot.msu.edu
Date: Sun, 6 Feb 2000 20:37:18 -0500 (EST)
Subject: Immnogoldlabeling on TEM

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Dear colleagues,
I am now currently using TEM to observe a filamentous structure
so-called hrp pilus produced by plant pathogen, Pseudomonas syringae which is
grown on planta mimic medium agar plates. Even though I could easily find this
structure through directly put the nickel-grid on the bacteria plate for a
certain time,followed by the negative staining, I can't convince other people
to believe it's the structure we are looking for as its size is around
7-10 nm, which is indistinguishable from that of bacterial flagellum.
So we want to label this structure by using the specific antibody against
its major structural protein followed by the 10 nm gold-conjugated second
antibody. The difficulty I am encountering now is most pili are washed away
after the whole immnuogoldlabeling process. Since I am a beginnner on TEM work,
I don't know whether it's a very commonly encountered problem in
immnogoldlabeling experiment or just a very special case happened to me, such
as the pilus i'm interested in is too vulnerable to survive the exclusive wash,
or I didn't master the whole immunogoldlabeling techinique. If it's a common
problem, I hope someone could kindly provide me with your valuable experience
on how to resolve this problem, how we can prevent them to be washed away. If
it's due to my techinical problem, could you please send me a very detailed
immunogoldlabeling protocol.
Any kind of help will be highly appreciated
have a nice week
yours
qiaoling jin
jinq-at-msu.edu








From: Augusto_A_Morrone-at-notes.seagate.com
Date: Mon, 7 Feb 2000 08:17:13 -0600
Subject: Re: TEM: Jeol 2010 question

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Jonathan, Andrew, Richard, Larry:

Thank you for your responses. Please note that I do perform the TILT and SHIFT
alignments (aka "pivot point alignment" in other instruments) on a regular
basis, but this doesn't solve the problem.

Regarding: "...What did you mean by a voltage centre? My interpretation of this
is that
the source high tension is wobbled and any shift in the (previously
focussed) image is minimised by tuning the beam tilts. This alignment would
be used for energy filtered TEM (EFTEM) when doing elemental mapping on a
Gatan Imaging Filter (GIF) for example".

This is a (standard) routine alignment for imaging as well. The voltage center
alignment is done by first focusing the specimen at the optimum OL current
(adjusting Z), then adjusting the Tilt (in image mode) and Shift (in diffraction
mode), and finally adjusting the beam tilts to minimize the image shift at the
center of the screen. The problem arises when the Brightness is changed, as is
typically adjusted when optimizing the illumination on the sample to record an
image (intense illumination) versus an SA diffraction pattern (fully overfocused
condenser). Apparently this brightness change affects the voltage center as
evidenced by a shift of the direct beam as that brightness change is effected in
the SAD mode. This shift in the direct beam is equivalent to a beam tilt.

Augusto






From: L. D. Marks :      ldm-at-apollo.numis.nwu.edu
Date: Mon, 7 Feb 2000 08:30:10 -0600 (CST )
Subject: Re: Parallel beam

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I regret to say that there are some important points which perhaps are
being missed. Classic optics, i.e. most TEM textbooks and introductory
optics texts are for one of two cases:
a) Every point in the condensor aperture (CA) is incoherent (i.e is not
related in phase in a statistical sense) with respect to any other point.
b) Every point in the CA is coherent (i.e. has a fixed phase) with
respect to any other point.

Approximation a) is used in most computer codes for HREM, approximation
b) for STEM.

Alas, both a) and b) are only approximations! Current microscopes,
particularly FEGs (but perhaps others as well) operate under conditions
when neither of the two approximations are valid. While the analytic
theory is well established (see the Chapter in Born and Wolff on
partial coherence), it is not easy and there are no clean solutions
that you can write down. (It can be solved numerically, but this
would require big 4-D FFT's which are beyond most current computers.)

The simplest test is to fully focus the illumination. If the spot
size is A nm, the coherence in the CA is about 1/A nm-1 (which you can
convert to an angle as appropriate). If this is small relative to the
CA radius you are working with case a) above. If it is about the same as the
CA radius you have b). If it is 2-3 times less, neither approximation
is correct!

The moral of the story:
1) If you have a), you can simulate an HREM image with confidence
using any of the available programs. They are WRONG quantitatively
otherwise.
2) If you have a), you can use simple "ray-diagram" optics,
but not otherwise.
3) If you have b) the simple equations for STEM operation are
valid, but not otherwise.

Sorry, but as microscopes improve the theory we need to understand them
can change.

++++++++++++++++++++++++++++++++++++++++++++++++
Laurence Marks
Department of Materials Science and Engineering
Northwestern University
fax: (847) 491-7820
mailto:l-marks-at-nwu.edu
http://www.numis.nwu.edu
++++++++++++++++++++++++++++++++++++++++++++++++





From: Michael Herron :      herro001-at-maroon.tc.umn.edu
Date: Mon, 07 Feb 2000 09:20:44 -0600
Subject: Diameter of human RBC?

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All,

I am trying to get a handle on the scale of some old pictures. There
are human RBC in theses pictures. Can anyone tell me the diameter of
your average human RBC?

Thanks
Mike

--
_________________________________________________
/ Michael J. Herron /
/ U of MN,Medicine/Infectious Diseases /
/ herro001-at-maroon.tc.umn.edu /
/ http://128.101.243.213 /
/_____________________________________________/




From: Walck. Scott D. :      walck-at-ppg.com
Date: Mon, 7 Feb 2000 10:52:29 -0500
Subject: Be X-ray peaks -I don't think so

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When a characteristic X-ray is given off from an atom, it carries 1 h-bar
(Planck's constant divided by 2 pi) of angular momentum. The electronic
transition that occurred for the X-ray to come off must conserve angular
momentum of the system. Therefore, a transition from a 2s1/2 to a 1s1/2
state is forbidden by selection rules. You can't produce a Be X-ray.
-Scott

Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)






From: A. K. Christensen :      akc-at-umich.edu
Date: Mon, 07 Feb 2000 11:38:52 -0500
Subject: Re: Diameter of human RBC?

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Mike: According to Fawcett's CONCISE HISTOLOGY (page 32), "In the blood,
erythrocytes are 8.5 micrometers in diameter, but in the dehydrated state,
in dried blood smears, they measure 7.6 micrometers." --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
Office: 2703A Medical Science II Building
Tel (work) (734) 763-1287, Fax (work) (734) 763-1166
E-mail: akc-at-umich.edu
http://www.umich.edu/~akc/
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
--On Mon, Feb 7, 2000 9:20 AM -0600 Michael Herron
{herro001-at-maroon.tc.umn.edu} wrote:

} All,
}
} I am trying to get a handle on the scale of some old pictures. There
} are human RBC in theses pictures. Can anyone tell me the diameter of
} your average human RBC?
}
} Thanks
} Mike
}
} --
} _________________________________________________
} / Michael J. Herron /
} / U of MN,Medicine/Infectious Diseases /
} / herro001-at-maroon.tc.umn.edu /
} / http://128.101.243.213 /
} /_____________________________________________/








From: donald j marshall :      dmrelion-at-world.std.com
Date: Mon, 7 Feb 2000 12:06:10 -0500 (EST)
Subject: Re: Computer lock-ups from static

Contents Retrieved from Microscopy Listserver Archives
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John, If they do, I hope that they will respond to the whole list because we
have clients who have experienced similar problems, especially re: computers
that are controlling VIS spectrometers collecting CL spectra. I know that
there are in line RF filters available (e.g., Steward Co.) and there may be
others but the final word is not in on how well these work yet.

Don Marshall


} From microprobe-at-microanalysis.org Mon Feb 7 11:43:28 2000
} }
} From: John Hunt {hunt-at-ccmr.cornell.edu}
} Subject: Computer lock-ups from static
} To: microprobe-at-microanalysis.org
}
}
} Dear Probe community,
} It is winter in Central NY, and the room humidity is 33%. Static
} electricity (I believe) has been causing frustrating computer lockups
} requiring rebooting our microprobe workstation.
} I am wondering if anyone has found the best solution for preventing
these
} problems.
}
}
}
}
} John Hunt
} Cornell Center for Materials Research
} 607-255-0108 microprobe
} -3789 SEM/TEM
} -2365 fax
}

Donald J. Marshall
Relion Industries
P.O. Box 12
Bedford, MA 01730
Ph: 781-275-4695
FAX: 781-271-0252
email dmrelion-at-world.std.com

Cathodoluminescence, mass spectroscopy, electron beam technology


"A weed is a flower out of place."

(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)




From: Marisa Ahmad :      mahmad-at-semiconductor.com
Date: Mon, 7 Feb 2000 12:33:38 -0500
Subject: tungsten filaments

Contents Retrieved from Microscopy Listserver Archives
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I would like to tap the wealth of information out there.

We are experiencing a lot of problems with the tungsten filaments that we
are currently using on our JEOL 5800 SEM. During long acquisitions (many
hours) there is a lot of brightness drift and shifting, even when lengthy
stabilization periods are given prior to the acquisitions. JEOL has
checked out the SEM itself, and can't find the source for the drifting. I
was thinking that it might be the filament. I was wondering if there are
different types of tungsten filaments out there, and which is the most
stable over long periods of time. I would like to try out all of them
(considering trying to switch our entire system to a LaB6 filament is a bit
much for now). Any recommendations or experiences you have would help us
out a lot.

Thanks in advance!


Marisa Ahmad
R&D Specialist
Semiconductor Insights
mahmad-at-semiconductor.com


weather report (for those interested):
It's unusually sunny and warm for here in Ottawa.....only -10 degrees C (but
it feels like -25 with the wind). It's such a nice day that I washed my
car this morning since it's not really supposed to be brown - now all the
doors and windows are frozen shut. {sigh}





From: Marie E. Cantino :      cantino-at-uconnvm.uconn.edu
Date: Mon, 7 Feb 2000 12:45:54 -0500
Subject: Re: Diameter of human RBC?

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In my experience, RBCs prepared for SEM by critical point drying or HMDS
shrink even more and may be only 4 or 5 microns across.

Marie
}
} Mike: According to Fawcett's CONCISE HISTOLOGY (page 32), "In the blood,
} erythrocytes are 8.5 micrometers in diameter, but in the dehydrated state,
} in dried blood smears, they measure 7.6 micrometers." --Kent
}

Dr. Marie E. Cantino
Dept. of Physiology and Neurobiology, U-2131
University of Connecticut
Storrs, CT 06269-2131
Phone: 860-486-3588
Fax: 860-486-1936






From: Karl E. Garsha :      keg-at-csd.uwm.edu
Date: Mon, 7 Feb 2000 12:03:51 -0600 (CST)
Subject: Re: Immnogoldlabeling on TEM

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------------- Begin Forwarded Message -------------



------------- End Forwarded Message -------------






From: Karl E. Garsha :      keg-at-csd.uwm.edu
Date: Mon, 7 Feb 2000 12:10:39 -0600 (CST)
Subject: Re: Diameter of human RBC?

Contents Retrieved from Microscopy Listserver Archives
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It depends on the fixation/dehydration parameters. Somewhere in the
nieghborhood of 4 microns, dependant on the angle of measurement and the angle
of the sample with respect to the camera.
} Date: Mon, 07 Feb 2000 09:20:44 -0600
} From: Michael Herron {herro001-at-maroon.tc.umn.edu}
} X-Accept-Language: en
} MIME-Version: 1.0
} To: Microscopy listserv {Microscopy-at-sparc5.Microscopy.Com}
} Subject: Diameter of human RBC?
} Content-Transfer-Encoding: 7bit
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America





From: Margaret Brannigan :      brannign-at-asrr.arsusda.gov
Date: Mon, 7 Feb 2000 13:25:52 -0500
Subject: TEM: What is Brandes dip?

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Hi,

Can anyone tell me what a Brandes dip is and how it is done? The only thing
I know is that it can be used to detect virus particles from plant sap but
I can't find any references on it.

Thank You!

Margaret

Margaret Dienelt

Plant Pathologist
Electron Microscopy Lab

Floral and Nursery Plants Research Unit
U.S. National Arboretum/Agricultural Research Service/USDA

B. 010A, Rm. 238, BARC-W
10300 Baltimore Avenue
Beltsville MD. 20705 USA

(301) 504-6097
Fax: (301) 504-5096






From: Hao Li :      haoli-at-eng.umd.edu
Date: Mon, 07 Feb 2000 13:56:57 -0500
Subject: Simulation of dislocation network

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Does anybody know any program capable of doing TEM simulation of
dislocation network, e.g., misfit dislocations in plane view TEM? I know
some programs can do one or two dislocations, but what I need is the
simulation for a set of dislocations.
Thanks a lot.

Hao Li





From: Steve Chapman :      PROTRAIN-at-CompuServe.COM
Date: Mon, 7 Feb 2000 13:59:53 -0500
Subject: Automatic Microscope and Coherence

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Hi

It has been very interesting reading about diffraction and electron beams
but it is quite clear that we often give the instrument credit, "the
instrument does it automatically", when credit is not due. Almost every
"automatic" is a designers estimate or better still a calculation, it is a
setting!

After 36 years with TEM I have to say that the only area that the
instrument could set in the SA mode could be the approximate position of
the first image plane. We normally move to the SA mode to free up the
diffraction lens (1st intermediate) to enable it to be focussed on the
diffraction (intermediate) aperture independent of the magnification
system. We would then focus the image inside the aperture with the
objective lens to ensure that the selected area IS the area in the
diffraction pattern. Without this procedure electron rotation and
misalignments could give you a diffraction pattern from an area not in the
centre of the screen! Users are correct in that if they set the eucentric
point or better still, adjust the specimen height to focus at the same
objective lens current value, that done the first image plane will be at
the same point within the diffraction (intermediate) lens.

So how could the Philips work? I would guess they set the diffraction lens
at approximately the first image plane. Then as they do not use it within
their lens scheme for the SA magnifications it is fixed at this focus, good
enough I think?

A second point, parallel beams are very important if you are chasing the
best image quality, biology or materials, most times we need high
coherence. There are misunderstandings on how to obtain a parallel beam or
high coherence, setting the final condenser underfocus is incorrect. The
procedure for high coherence would be to use the smallest spot size you can
tolerate (this probably means you must up the emission current). Once in
this condition overfocus the final condenser (clockwise from crossover) the
spot, what ever size it is now, becomes your virtual source. The further
overfocus you go the greater the beam coherence and that is what we are
after. You will deduce the smaller the condenser aperture the sharper the
spot and the smaller the spot the greater the coherence for a given degree
of overfocus.

Work with a design team and they expect everyone to overfocus, they expect
everyone to use small condenser apertures and they expect everyone to use
small spot sizes. They do not expect everyone to use too low an emission
current (almost everyone does, we have talked before about filament life
being the most important feature of many laboratories) because this makes
the task too difficult!
There must be people more knowledgeable than I who will cut out all the
guesswork?

Steve Chapman
Senior Consultant
Protrain
Electron microscopy courses world wide
http://www.emcourses.com
Tel & Fax 44 (0) 1280 814774




From: Kalman Rubinson :      kr4-at-is2.nyu.edu
Date: Mon, 7 Feb 2000 14:27:14 -0500 (EST)
Subject: Re: Diameter of human RBC?

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On Mon, 7 Feb 2000, Michael Herron wrote:

} I am trying to get a handle on the scale of some old pictures. There
} are human RBC in theses pictures. Can anyone tell me the diameter of
} your average human RBC?

When I was a lad...... in grad school, we were told it was
a useful reference at 7um diameter.

Kal





From: Joseph Passero :      jp-at-spacelab.net
Date: Mon, 07 Feb 2000 15:15:13 -0500
Subject: ANNOUNCEMENT Course on Polarized Light Microscopy

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New York Microscopical Society

Bernard Friedman Memorial Workshop

Polarized Light Microscopy

Two consecutive weekends

Saturday, April 21, Sunday, April 22, 2000
Saturday, April 28, Sunday, April 29, 2000

An advanced course on polarized light microscopy which will cover the following topics:
The nature of polarized light
The origin and interpretation of interference colors
Birefringence and crystal orientation
The Indicatrix
Compensation and variable compensators
Interference figures and their interpretation

The workshop will consist of two consecutive weekends of lectures and hands on labs to cover
the theoretical and practical aspects of polarized light microscopy.

The course instructors include;

Jan Hinsch of Leica, Inc.,

John Reffner of Trace Consulting,

N.Y.M.S. Instructor Donald O'Leary.

WHEN: April 22, 23, 28 & 29, 2000 from 10 A.M. to 4 P.M.

WHERE: Location To be Announced.

COST: $275 for N.Y.M.S. members, $295 for non-members (includes membership)
Lunch and course materials are included. Checks made out to N.Y.M.S.

WHO: advanced course for those who have completed "The Use of the Microscope" or
are experienced in microscopy and familiar with the theory of its use.

HOW: Register using the form below. Limited to the first 12 registrants.


FURTHER INFORMATION: Contact Donald O'Leary

eMail: mailto:donoleary-at-worldnet.att.net

(201) 797-8849 Voice Phone Number

PLEASE POST
------------------------------------------------------------------------------------------------------------

Return form to Donald O'Leary, 6 Chittenden Road, Fair Lawn, NJ 07410.

Registration Form

Polarized Light Microscopy, April 22, 23, 28 & 29, 2000

N.Y.M.S. Member _________________ ($275.00) Non-Member __________($295.00)

Name ___________________________________________________________________________

Address _________________________________________________________________________

City _________________________________ State __________________ Zip Code _______________

Phone (W) ____________________________ (H) ___________________________

eMail Address: ______________________________________




________________________________________________________
1stUp.com - Free the Web
Get your free Internet access at http://www.1stUp.com




From: Robert Underwood :      underwoo-at-u.washington.edu
Date: Mon, 7 Feb 2000 13:23:54 -0800 (PST)
Subject: Re: Cy5 filtercube today on a upright flourescent microscope

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Hi Patricia,

I'm just taking a guess. Is it possible, depending on the type of filters
used in the cube, that infrared is passing through as flare if your first
excitation filter is not blocking the IR? Or there may be a coating
problem on your filter? We have an IR blocking filter right after the
mercury lamp. The CCD cameras are extremely sensitive to IR and this has
caused problems on our system. But in your situation I don't know why it
would pass through. So it is probably something else.

Bob
Morphology core
U of Washington

On Fri, 4 Feb 2000, Patricia A. Glazebrook 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 tried a Cy5 filtercube today on a upright flourescent microscope equiped
} with a Diagnostics Instrument Spot Camera. My tissue had been
} immunolabelled with Cy5 and I did not expect to see thru the microscope. So
} we took pictures with the Spot Camera and the immunolabeling seemed to
} work. The problem was that there was uneven illumination in the pictures,
} half moon of brightness and the rest of the field dark. The salesperson did
} check that the mercury bulb was aligned correctly. Also there was no
} ambient light from the room causing the shadow.
}
} Anyone with an idea  of what is causing this problem?
}
} Thank-you, Pat Glazebrook
}
}
}
}
}





From: Barbara Foster :      mme-at-map.com
Date: Mon, 07 Feb 2000 16:26:47 -0500
Subject: Re: Help:A comparison of optics

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One more thought to support Gary's observation about the importance of the
NA of the condenser: The full equation for the limit of resolution is R =
(1.22)(wavelength)/(NAobjective + NAcondenser). However, if you don't oil
it to the back of the slide, you might as well use a lower NA condenser.
Reminds me of the hematology scope built by an otherwise sensible company,
meant to be used in ultra high resolution applications. The manufacturer
was very good about providing a quality, high NA, oil immersion objective
as well as a high NA condenser. The only problem was that you could never
raise the condenser high enough to oil it to the back of the slide and,
therefore, use it at the NA inscribed... only at about .95! What a waste
of time and money!

Also, a reminder that the NAs engraved are the maximum working limit.
Closing down an aperture iris or an iris in the back focal plane of the
objective reduces the NA accordingly.

One last fact: the NA affects both resolution AND edge information, so,
for the crispest, highest resolution images, go for high NA in both
objective and condenser.

Best regards,
Barbara Foster
Consortium President
Microscopy/Microscopy Education ...Educating microscopists for greater
productivity.

125 Paridon Street Suite 102 Springfield, MA 01118
PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Visit our web site {http://www.MME-Microscopy.com/education}


At 03:02 PM 2/4/00 -0600, Dr. Gary Gaugler wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America




From: William Tivol :      tivol-at-wadsworth.org
Date: Mon, 7 Feb 2000 16:35:56 -0500 (EST)
Subject: Re: tungsten filaments

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Marisa Ahmad:
} I would like to tap the wealth of information out there.
}
} We are experiencing a lot of problems with the tungsten filaments that we
} are currently using on our JEOL 5800 SEM. During long acquisitions (many
} hours) there is a lot of brightness drift and shifting, even when lengthy
} stabilization periods are given prior to the acquisitions. JEOL has
} checked out the SEM itself, and can't find the source for the drifting. I
} was thinking that it might be the filament. I was wondering if there are
} different types of tungsten filaments out there, and which is the most
} stable over long periods of time. I would like to try out all of them
} (considering trying to switch our entire system to a LaB6 filament is a bit
} much for now). Any recommendations or experiences you have would help us
} out a lot.
}
Dear Marisa,
Do you have control over the filament voltage? If so, set the
voltage just a hair over that necessary to saturate the filament. If
the voltage is too low, parts of the filament will not emit electrons
which get through the wehnelt and into the beam; if too high, there may
be significant evaporation of the filament; either may cause drifting
and/or shifting. I have had very good luck with the stabilities of
both Agar and Ebtec (now Energy Beam Sciences) filaments; not that
others may not be as good, I haven't tried them. Ebtec makes several
shapes of filament, and one may be best for long-term stability. I
have no interest in Agar or Ebtec except as a satisfied customer.
Yours,
Bill Tivol




From: Barbara Foster :      mme-at-map.com
Date: Mon, 07 Feb 2000 17:07:29 -0500
Subject: Re: Computer lock-ups from static

Contents Retrieved from Microscopy Listserver Archives
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John,

Have you considered a "local" remedy, like putting up a humidifier? I know
that static can be an issue, but I have never heard of it being this bad
before.
In more humid environments, we often put a small enclosure around a light
microscope then put out a dish of Drierite. Perhaps some simple plastic
sheeting, like they sell for exterior storm windows/wind breaks, can be
hung from the ceiling around your microprobe plus the humidifier will keep
the humidity immediately around your microprobe more balanced.

Best of luck on a tough problem.

Barbara Foster
Consortium President
Microscopy/Microscopy Education ...Educating microscopists for greater
productivity.

125 Paridon Street Suite 102 Springfield, MA 01118
PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Visit our web site {http://www.MME-Microscopy.com/education}


At 12:06 PM 2/7/00 -0500, donald j marshall wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America




From: BLADE1691-at-aol.com
Date: Mon, 7 Feb 2000 17:33:41 EST
Subject: pictures

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---------- Forwarded message ----------


how can i get a picture of you seal




From: G. Macdonald :      glenmac-at-u.washington.edu
Date: Mon, 7 Feb 2000 14:52:34 -0800 (PST)
Subject: Re: Cy5 filtercube today on a upright flourescent microscope

Contents Retrieved from Microscopy Listserver Archives
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Is this a Spot Jr. or a SpotII? The IR filter in the Jr. cuts off at 650
nm, the SpotII puts it on a slider. Is it all the way out or [partially
occluding the image? Bob Fern
caught a lot of the possible issues, but is your lamp properly focussed
and aligned? Are you picking up light through the eyepieces? We have a
Nikon that will show a semi-circle of light in from a
recessed ceiling light
that projects to the scope at an angle.

Regards,
Glen


Glen MacDonald
Research Scientist
Hearing Research Laboratories of the
Virginia Merrill Bloedel Hearing Research Center
Box 35-7923
University of Washington
Seattle, WA 98195-7923
(206) 616-4156
glenmac-at-u.washington.edu


On Fri, 4 Feb 2000, Patricia A. Glazebrook 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 tried a Cy5 filtercube today on a upright flourescent microscope equiped
} with a Diagnostics Instrument Spot Camera. My tissue had been
} immunolabelled with Cy5 and I did not expect to see thru the microscope. So
} we took pictures with the Spot Camera and the immunolabeling seemed to
} work. The problem was that there was uneven illumination in the pictures,
} half moon of brightness and the rest of the field dark. The salesperson did
} check that the mercury bulb was aligned correctly. Also there was no
} ambient light from the room causing the shadow.
}
} Anyone with an idea  of what is causing this problem?
}
} Thank-you, Pat Glazebrook
}
}
}
}
}






From: Michael Bode :      mb-at-soft-imaging.com
Date: Mon, 7 Feb 2000 16:06:21 -0700
Subject: RE: Computer lock-ups from static

Contents Retrieved from Microscopy Listserver Archives
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Out here in the dry West (here we have Summer humidity around the 33%
value you mention, in Winter we go a lot further down) we have the same
problem. There are a few methods to help:

1) increase humidity. Perhaps you can use a humidifier in your equipment
room?
2) prevent charging. There are shoes available that prevent charging.
3) get rid of charging. There are several options here:

a) The easiest way and what I usually do if no other method is
available, is to touch some metal part NOT of the PC but a chair or
table before touching the PC. This requires a) that a suitable object is
near the PC, b) that you're not afraid of "shocking" yourself, and c)
that you don't move around a lot while working on the PC.

b) Purchase a grounding mat to place in front of the equipment. these
mats are connected to the AC ground (use the same as the PC ground). If
the mat is big enough so that everybody has to step on the mat first AND
is wearing the conductive shoes, everything should be OK.

c) wear grounded wrist straps. These require of course that you put them
on.

We use a combination of these techniques to protect our equipment both
in the office and the production floor (believe me, it IS necessary
here). As bad as it is for electronic equipment, though, it is just
great for comfort levels outside. 90 degrees (F) feel just right for
bicycling, climbing, rafting...

One company where you can buy this stuff is called "-at-once". Their web
site is www.4atonce.com. I just happened to find their catalog first.
There are many other companies out there that sell static control
equipment. We have no financial interest in this company whatsoever.

Michael

Michael Bode, Ph.D.
Soft Imaging System Corp.
1675 Carr St., #105N
Lakewood, CO 80215
===================================
phone: (888) FIND SIS
(303) 234-9270
fax: (303) 234-9271
email: mailto:info-at-soft-imaging.com
web: http://www.soft-imaging.com
===================================



} ----------
} From: donald j marshall[SMTP:DMRELION-at-WORLD.STD.COM]
} Sent: Monday, February 07, 2000 10:06:10 AM
} To: hunt-at-ccmr.cornell.edu
} Cc: Microscopy-at-sparc5.Microscopy.Com
} Subject: Re: Computer lock-ups from static
} Auto forwarded by a Rule
}
------------------------------------------------------------------------
The Microscopy ListServer -- Sponsor: The Microscopy Society of America



John, If they do, I hope that they will respond to the whole list
because we
have clients who have experienced similar problems, especially re:
computers
that are controlling VIS spectrometers collecting CL spectra. I know
that
there are in line RF filters available (e.g., Steward Co.) and there may
be
others but the final word is not in on how well these work yet.

Don Marshall


} From microprobe-at-microanalysis.org Mon Feb 7 11:43:28 2000
} }
} From: John Hunt {hunt-at-ccmr.cornell.edu}
} Subject: Computer lock-ups from static
} To: microprobe-at-microanalysis.org
}
}
} Dear Probe community,
} It is winter in Central NY, and the room humidity is 33%.
Static
} electricity (I believe) has been causing frustrating computer lockups
} requiring rebooting our microprobe workstation.
} I am wondering if anyone has found the best solution for
preventing
these
} problems.
}
}
}
}
} John Hunt
} Cornell Center for Materials Research
} 607-255-0108 microprobe
} -3789 SEM/TEM
} -2365 fax
}

Donald J. Marshall
Relion Industries
P.O. Box 12
Bedford, MA 01730
Ph: 781-275-4695
FAX: 781-271-0252
email dmrelion-at-world.std.com

Cathodoluminescence, mass spectroscopy, electron beam technology


"A weed is a flower out of place."

(Please note: Do not send email with attachments to this address.
Instead, send it to donbarlen-at-aol.com. Thank you.)




From: JoAnn Buchanan :      redhair-at-leland.Stanford.EDU
Date: Mon, 7 Feb 2000 17:24:50 -0600
Subject: GFP antibodies

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I am planning to try EM immunolabelling with GFP antibodies. Does anyone
out there know of the best source for GFP antibodies and had any luck with
using them for TEM. I would like to try pre-embedding with a nanogold
labelled secondary. Any suggestions would be appreciated. Thank you.

JoAnn Buchanan
Molecular and Cellular Physiology
Stanford University School of Medicine
Stanford, CA 94305
650-723-5856






From: Frederick Schamber :      fhscham-at-sgi.net
Date: Mon, 07 Feb 2000 19:04:05 -0500
Subject: Re: tungsten filaments

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Marisa,

Just a quick comment for whatever it's worth. Your posting suggests that you
are concerned about mechanical drifts of the filament -- the subject of a
certain body of folklore in the microscopy community. It's been my experience
that the principal drift in a tungsten filament is not the filament itself, but
rather it is the mechanism supporting the filament which drifts.

The actual issue with drifting filaments is that mechanical stresses in the wire
will gradually relieve under heat, causing the filament to distort slightly.
This is a first-time-used issue. However, I believe most manufacturers include
an annealing step in their process to remove these stresses before the filament
is shipped to a customer (at least, that's what we used to do in another company
when we manufactured our own filaments). In any case, the filament is operated
at a very high temperature and it doesn't take long to anneal out these stresses
under normal use, so long-term drift effects shouldn't be an issue.

My experience is that long-term mechanical drifts originate elsewhere in the gun
mechanism. When you think about it, this makes sense. The filament itself has
very little mass and quickly reaches its ultimate operating temperature
distribution. However, the rest of the gun assembly is much more massive and,
depending on various design considerations, may take hours to reach thermal
equilibrium. Meanwhile, all of those intricate parts are expanding -- and many
at different rates. Principal suspects, in my opinion, are lateral adjusting
and clamping screws -- for example, a set of radial set screws is commonly used
to center the ceramic base of the filament within a ring of some type. I used
to work with a rather complex assembly which positioned the filament via lateral
screws and sometimes had episodes of "filament drift". Subsequently, I have had
nearly a decade of experience with a very simple design which supports the
filament solely via axial pressure and find that "filament drift" isn't an
issue. My conclusion is that a lot of what I used to call "filament drift" was
actually mechanism drift. So if you are experiencing mechanical instabilities
of the filament, I would suspect the way the filament is being supported more
than the filament itself.

This opinion is based on my own experiences, however, and I will be very
interested to hear if others have had different experience.

Fred Schamber
RJ Lee Instruments Limited

Marisa Ahmad 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 would like to tap the wealth of information out there.
}
} We are experiencing a lot of problems with the tungsten filaments that we
} are currently using on our JEOL 5800 SEM. During long acquisitions (many
} hours) there is a lot of brightness drift and shifting, even when lengthy
} stabilization periods are given prior to the acquisitions. JEOL has
} checked out the SEM itself, and can't find the source for the drifting. I
} was thinking that it might be the filament. I was wondering if there are
} different types of tungsten filaments out there, and which is the most
} stable over long periods of time. I would like to try out all of them
} (considering trying to switch our entire system to a LaB6 filament is a bit
} much for now). Any recommendations or experiences you have would help us
} out a lot.
}
} Thanks in advance!
}
} Marisa Ahmad
} R&D Specialist
} Semiconductor Insights
} mahmad-at-semiconductor.com
}
} weather report (for those interested):
} It's unusually sunny and warm for here in Ottawa.....only -10 degrees C (but
} it feels like -25 with the wind). It's such a nice day that I washed my
} car this morning since it's not really supposed to be brown - now all the
} doors and windows are frozen shut. {sigh}





From: schmutzm-at-lear.u-strasbg.fr (Schmutz Marc)
Date: Tue, 8 Feb 2000 09:27:07 +0900
Subject: Hematology Slide Stainer

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Hi Colleagues around the world,



We're interested in knowing a little bit more about the Hematology Slide
Stainer RSG 61 commercialized by EMS. So if you have one can you please
share your experience with us. Give the plus and also all the minus. If you
have another equivalent brand in which you're fully satisfied of course say
it. Naturally salesman can contact me (offline only please, so the whole
community will not be annoided...)

Thank you for your help


Marc


PS hi Keith how is weather in Plymouth ? Here it's raining and we're singing...





------------------------------
SCHMUTZ Marc
IGBMC
1 rue Laurent FRIES
BP 163
F 67404 Illkirch Cedex
FRANCE

Tel: +33 (0)388 653 330 direct
Fax: +33 (0)388 653 201
email:schmutzm-at-lear.u-strasbg.fr

------------------------------






From: Bart Cannon :      cannonmp-at-accessone.com
Date: Mon, 07 Feb 2000 16:39:50 -0800
Subject: Solid State Electron Emitters

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Donald,

Nasa has been working on micro-machined solid state electron emitters
and Thomas George of that agency has published a design utilizing an
array of them as the source for a "palm top" SEM in the August, 1998
issue of Nasa Tech Briefs. The proposed SEM relies upon the speculated
use of an electon transparent membrane at the bottom of the lens column
in this sealed, vacuum pumpless system. Resolution is anticipate in the
micron range.

Nasa has also published designs for miniature x-ray tubes in an October
1995 issue of Nasa Tech Briefs.

As far as I know a solid state high energy electron gun exists only as a
Star Wars project.

Bart Cannon
Cannon Microprobe
Seattle
206 522 9233




From: Bart Cannon :      cannonmp-at-accessone.com
Date: Mon, 07 Feb 2000 16:39:50 -0800
Subject: Solid State Electron Emitters

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Donald,

Nasa has been working on micro-machined solid state electron emitters
and Thomas George of that agency has published a design utilizing an
array of them as the source for a "palm top" SEM in the August, 1998
issue of Nasa Tech Briefs. The proposed SEM relies upon the speculated
use of an electon transparent membrane at the bottom of the lens column
in this sealed, vacuum pumpless system. Resolution is anticipate in the
micron range.

Nasa has also published designs for miniature x-ray tubes in an October
1995 issue of Nasa Tech Briefs.

As far as I know a solid state high energy electron gun exists only as a
Star Wars project.

Bart Cannon
Cannon Microprobe
Seattle
206 522 9233




From: Bart Cannon :      cannonmp-at-accessone.com
Date: Mon, 07 Feb 2000 16:39:50 -0800
Subject: Solid State Electron Emitters

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Donald,

Nasa has been working on micro-machined solid state electron emitters
and Thomas George of that agency has published a design utilizing an
array of them as the source for a "palm top" SEM in the August, 1998
issue of Nasa Tech Briefs. The proposed SEM relies upon the speculated
use of an electon transparent membrane at the bottom of the lens column
in this sealed, vacuum pumpless system. Resolution is anticipate in the
micron range.

Nasa has also published designs for miniature x-ray tubes in an October
1995 issue of Nasa Tech Briefs.

As far as I know a solid state high energy electron gun exists only as a
Star Wars project.

Bart Cannon
Cannon Microprobe
Seattle
206 522 9233




From: Barbara Foster :      mme-at-map.com
Date: Mon, 07 Feb 2000 19:57:13 -0500
Subject: LM: Course reminder

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Just a friendly reminder about the upcoming American Chemical Society
Course, "Applied Optical Microscopy". This course is not just for chemists!

If you need a refresher on a basic technique, need to know more about
interpreting images gathered by a variety of contrast systems (including
Hoffman Modulation Contrast and Polarized light), or are moving more into
digital imaging, this is a great course for you!

Hands-on, 3 days of total immersion, basic principles through advanced
techniques, quantitative polarized light.... and New Orleans, thrown in for
good measure! $895 for ACS members; $995 for non-members (tuition and
materials only).

Visit the MME website for details: {http://MME-Microscopy.com/education}

Best regards,
Barbara Foster
ACS Course Coordinator

Microscopy/Microscopy Education ...Educating microscopists for greater
productivity.

125 Paridon Street Suite 102 Springfield, MA 01118
PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Visit our web site {http://www.MME-Microscopy.com/education}
******************************************************




From: Hay, Karen :      KHay-at-ahs.llumc.edu
Date: Mon, 7 Feb 2000 19:13:42 -0800
Subject: RE: Diameter of human RBC?

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---------- Forwarded message ----------



-----Original Message-----
From: Michael Herron [SMTP:herro001-at-maroon.tc.umn.edu]

I am trying to get a handle on the scale of some old pictures.
There
are human RBC in theses pictures. Can anyone tell me the diameter
of
your average human RBC?


Mike-

I have recently subscribed to this list in the hopes of picking up some SEM
pointers as I am beginning to dip my toes into the field. I certainly
can't answer any SEM questions, but I CAN answer your question regarding the
size of human RBCs. On a dried film, normal RBC's have a diameter of
between 7.2-7.9 um. In certain disease states, they can deviate markedly
from this. Microcytic RBC's can be well under 6 um, while macrocytes will
have diameters greater than 9 um.

Good luck in your sizing!

{ {...} }
Karen Hay, MS, MT(ASCP)
Hematology Research, MC 2522
Loma Linda University Medical Center
Loma Linda, CA 92354
Tel: (909) 558-4000 x45350
Fax: (909) 558-4189
Khay-at-ahs.llumc.edu {mailto:Khay-at-ahs.llumc.edu}






From: COURYHOUSE-at-aol.com
Date: Mon, 7 Feb 2000 23:24:15 EST
Subject: Re: Diameter of human RBC?

Contents Retrieved from Microscopy Listserver Archives
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what is a rbc?
in machining we had a theoretical size referred to by the Dimension of a RCH
thanks Ed Sharpe archivist for SMECC




From: Ken Tiekotter :      tiekotte-at-up.edu
Date: Mon, 7 Feb 2000 22:45:24 -0800 (PST)
Subject: Re: Diameter of human RBC?

Contents Retrieved from Microscopy Listserver Archives
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Michael,
How about ~7.5 to 8.5um in diameter (depending on fixation/preparation)
and ~2um in greatest thickness.
-Ken

On Mon, 7 Feb 2000, 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,
}
} I am trying to get a handle on the scale of some old pictures. There
} are human RBC in theses pictures. Can anyone tell me the diameter of
} your average human RBC?
}
} Thanks
} Mike
}
} --
} _________________________________________________
} / Michael J. Herron /
} / U of MN,Medicine/Infectious Diseases /
} / herro001-at-maroon.tc.umn.edu /
} / http://128.101.243.213 /
} /_____________________________________________/
}
}





From: Bright, Alan :      ABright-at-brightinstruments.com
Date: Tue, 8 Feb 2000 08:59:57 -0000
Subject: SEM: automotive paints

Contents Retrieved from Microscopy Listserver Archives
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Dear Jorgelina,

We have supplied a number of Cryostats & Microtomes to the automotive
industry for sectioning paint finishes on steel body parts & plastic
components e.g. bumper (fenders) door handles, door handle surrounds
etc. Please view our Website to see our range of instruments.

If I can be of further assistance please come back to me.

Best Regards

Alan Bright

Bright Instrument Co.Ltd.
St Margaret's Way
Huntingdon
PE18 6EB
England

Tel No:+44 (0)1480 454528
Fax No:+44 (0)1480 456031
Email: AlanBright-at-brightinstruments.com
Web Site: www.brightinstruments.com



-----Original Message-----
} From: "csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com
[mailto:"csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com]
Sent: 08 February 2000 17:35
To: MICROSCOPY-at-sparc5.microscopy.com


Hi!
My name is Jorgelina C. Altamirano. I'm an undegrated student of
the
licenciate in chemical sciences.
I'm currently preparing my thesis, being its subjet: The
morphological
characterization and the chemical analysis of automotive paints with
forensic
purposes.
I'm undergoing my work using SEM and EPMA techniques with a
JEOL,
JSM 35C and a microprobe EDAX PV9500.
I've chosen some papers as references:
- Beam et al, Analysis Protocol for discrimination for
automotive
paints by SEM-EDAX, Journal of Forensic Sciences, 35, 5, 1990
- Zieba-Palus et al, Mikrochim. Acta, 14, 1997.

I've taken same samples of automotive paints (such as Fiat,
Peugeot,
Renault, Ford, Chevrolet, Volkswagen) corresponding to the years
1994-99.
I've found many difficulties while preparing them in:
- separating the paint from the body work of the car;
- cutting the specimen;
- to stick specimen to the graphite specimen pedestal;

Though I've made the primary morphological and chemical analysis
and I
haven't observed great differences between the diferent automotive
paints
trades (while polyuretane paints and belayer metalizer paints already
tested),
but I'd found great differences between layers and layers of the paints
specimen.
As I'm not quite satisfied with the results obtained to be used
for
forensic purposes, I would greatly appreciate making contacts with
specialist
fo this subject, so as to receive any comments on this subject.
Thank you in advance for your help.
Jorgelina Altamirano
CERIDE,
Guemes 3450
(3000) Santa Fe, Argentina
e-mail: csedax-at-arcride.edu.ar




From: mauty-at-MOOREPARK.TEAGASC.IE
Date: Tue, 08 Feb 2000 11:22:33 +0000
Subject: HeNe laser

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Jorgelina
SEM/EDS//EPMA is only one class of analytical methods that the Forensic
community applies to paint analysis for comparison and association with
original source. Although used routinely, probe methods are not used
exclusively. The organic components in paints are generally more
discriminating.
Sample preparation usually entails some form of embedment and either
microtomy or polishing in order to reveal internal structure.
Removal of paint from an auto body is tricky, and requires practice.The
manufacturers have designed their paint systems to prevent removal!
I would be glad to provide you with additional resources.

Dennis

_________________________________________________________________________
Dennis C . Ward voice: 202-324-2982
FBI Laboratory fax: 202-324-4018
Microanalysis Laboratory email:
DCWard-at-concentric.net


----- Original Message -----
} From: {"csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com}
To: {MICROSCOPY-at-sparc5.Microscopy.Com}
Sent: Tuesday, February 08, 2000 9:35 AM


Jorgelina
SEM/EDS//EPMA is only one class of analytical methods that the Forensic
community applies to paint analysis for comparison and association with
original source. Although used routinely, probe methods are not used
exclusively. The organic components in paints are generally more
discriminating.
Sample preparation usually entails some form of embedment and either
microtomy or polishing in order to reveal internal structure.
Removal of paint from an auto body is tricky, and requires practice.The
manufacturers have designed their paint systems to prevent removal!
I would be glad to provide you with additional resources.

Dennis

_________________________________________________________________________
Dennis C . Ward voice: 202-324-2982
FBI Laboratory fax: 202-324-4018
Microanalysis Laboratory email:
DCWard-at-concentric.net




----- Original Message -----
} From: "Bright, Alan" {ABright-at-brightinstruments.com}
To: {"csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com} ;
{MICROSCOPY-at-sparc5.Microscopy.Com}
Sent: Tuesday, February 08, 2000 12:59 AM


Jorgelina
SEM/EDS//EPMA is only one class of analytical methods that the Forensic
community applies to paint for characterization and association with
original source. Although used routinely, probe methods are not used
exclusively. The organic components in paints are generally more
discriminating.
Sample preparation usually entails some form of embedment and either
microtomy or polishing in order to reveal layer structure.
Removal of paint from an auto body is tricky, and requires practice.The
manufacturers have designed their paint systems to prevent removal!
I would be glad to provide you with additional resources.

Dennis

_________________________________________________________________________
Dennis C . Ward voice: 202-324-2982
FBI Laboratory fax: 202-324-4018
Microanalysis Laboratory email:
DCWard-at-concentric.net




----- Original Message -----
} From: {"csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com}
To: {MICROSCOPY-at-sparc5.Microscopy.Com}
Sent: Tuesday, February 08, 2000 9:35 AM


Dear all

I am planning to replace my Kr/Ar laser on a Zeiss 310 CSLM with two
separate lasers: 488nm Ar ion and HeNe 543nm. Does anyone have
experience of using a HeNe 543? I need this or something similar to
excite Rhodamine for food structure applications. I'd appreciate any
advice.

The reason I'm replacing the mixed gas laser is that has
averaged 25% downtime over the past three years.

Mark

Mark Auty
Dairy Products Research Centre
Moorepark, Fermoy, Co. Cork, Ireland.
mauty-at-moorepark.teagasc.ie
tel 00353 2542447
fax 00353 2542340






From: Stan Rice :      srice-at-alpha.utampa.edu
Date: Tue, 08 Feb 2000 09:04:43 -0400
Subject: LKB Nova

Contents Retrieved from Microscopy Listserver Archives
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Dear all,

Thank you for the many responses to my inquery about operation
instructions for the LKB Nova ultramicrotome. For future reference, I
have the instrument and the original instructions for the LKB Reichert
ultramicrotome if anyone has acquired one of these without instructions.
Thanks again for all of your responses.

Stan Rice
University of Tampa





From: Cap'n Peachfuzz :      bsteffen-at-arches.uga.edu
Date: Tue, 8 Feb 2000 08:42:57 -0500 (Eastern Standard Time)
Subject: Re: tungsten filaments

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On Mon, 7 Feb 2000 12:33:38 -0500 Marisa Ahmad
{mahmad-at-semiconductor.com} wrote:

} We are experiencing a lot of problems with the tungsten filaments that
} we are currently using on our JEOL 5800 SEM. During long acquisitions
} (many hours) there is a lot of brightness drift and shifting, even when
} lengthy stabilization periods are given prior to the acquisitions.
} JEOL has checked out the SEM itself, and can't find the source for the
} drifting

Marisa,

You can easily enough to see if it is filament drift. First, start the
filament alignment program to ensure alignment. Then with the beam on,
wait a few minutes for the filament to drift, then switch off the
autobeam function, and manually check the alignment using the 2
filament alignment knobs. If you can make the image brighter, then
there is filament drift. With my experiences with the 5800 though, I
suspect this is not the case.


W. L. (Buddy) Steffens, Ph.D
Dept. of Pathology
University of Georgia






From: Cap'n Peachfuzz :      bsteffen-at-arches.uga.edu
Date: Tue, 8 Feb 2000 09:02:01 -0500 (Eastern Standard Time)
Subject: Re: tungsten filaments

Contents Retrieved from Microscopy Listserver Archives
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On Mon, 7 Feb 2000 16:35:56 -0500 (EST) William Tivol
{tivol-at-wadsworth.org} 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 Marisa, Do you have control over the filament voltage? If so,
} set the voltage just a hair over that necessary to saturate the
} filament. If the voltage is too low, parts of the filament will not
} emit electrons which get through the wehnelt and into the beam; if too
} high, there may be significant evaporation of the filament; either may
} cause drifting and/or shifting. I have had very good luck with the
} stabilities of both Agar and Ebtec (now Energy Beam Sciences)
} filaments; not that others may not be as good, I haven't tried them.
} Ebtec makes several shapes of filament, and one may be best for
} long-term stability. I have no interest in Agar or Ebtec except as a
} satisfied customer. Yours,
} Bill Tivol


Marisa,

This is a good point that Bill makes...if you are slightly
undersaturated, there will be fluctuations in the rate of electron
emission from the filament. The 5800 automatically aligns and
saturates the filament when the Autobeam mode is selected...and on our
5800, saturation in this mode is a bit low. If you turn off the
autobeam switch and use the filament emission knob to slightly increase
the emission current, this might solve the problem at a cost of only a
small decrease in filament life.



W. L. (Buddy) Steffens, Ph.D
Dept. of Pathology
University of Georgia





From: Huggins, Bradley J :      HUGGINBJ-at-BP.com
Date: Tue, 8 Feb 2000 09:11:56 -0500
Subject: Re: Computer lock-ups from static

Contents Retrieved from Microscopy Listserver Archives
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John Hunt wrote:
}
} Dear Probe community,
} It is winter in Central NY, and the room humidity is 33%. Static
} electricity (I believe) has been causing frustrating computer lockups
} requiring rebooting our microprobe workstation.
} I am wondering if anyone has found the best solution for preventing
these
} problems.
}
John we had this same problem for many years while we were using the old
RT-11 based Tracor Northern and Kevex x-ray analyzers. The problem was
especially bad with the TN-5500. This was due to the keyboard with built in
scroll knob. The motion of turning that knob on the keyboard was perfect
for generating a little static. (This is pre- GUI and mouse days, except
for the Mac users!) Because of the low humidity conditions and severe
static accumulation, in winter months we were forced to set the entire
keyboard on a grounded pad. We also attached an antistatic strip to the
front of the keyboard. Because of the nature of the job, and the movement
to and from the computer and the microscope controls, we could not wear the
wrist straps. But with the conductive grounding strip across the entire
front of the keyboard we found that we could very easily get into the habit
of grounding our wrist or thumb on the strip before touching the rest of the
computer or keyboard. Then as we used the keyboard we would make some
effort to touch the grounding strip from time to time. This virtually
eliminated the problem. By the way I believe it was such a problem that
Tracor engineered a static discharge switch into the keyboard connection.


Brad Huggins
BPAmoco,
Naperville, Microscopy and Microanalysis




From: Leona Cohen-Gould :      lcgould-at-mail.med.cornell.edu
Date: Tue, 8 Feb 2000 12:44:34 -0400
Subject: Re: Diameter of human RBC?

Contents Retrieved from Microscopy Listserver Archives
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At 9:20 AM -0600 2/7/0, Michael Herron wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

Leona Cohen-Gould, M.S.
Sr. Staff Associate
Director, Electron Microscopy Core Facility
Manager, Confocal Microscopy Core Facility
Joan & Sanford I. Weill Medical College
of Cornell University
voice (212)746-6146
fax (212)746-8175






From: csedax-at-alpha.arcride.edu.ar
Date: Mon, 7 Feb 2000 17:36:17 -2359
Subject: SEM: automotive paints

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hi!
My name is Jorgelina C. Altamirano. I'm an undegrated student of the
licenciate in chemical sciences.
I'm currently preparing my thesis, being its subjet: The morphological
characterization and the chemical analysis of automotive paints with forensic
purposes.
I'm undergoing my work using SEM and EPMA techniques with a JEOL,
JSM 35C and a microprobe EDAX PV9500.
I've chosen some papers as references:
- Beam et al, Analysis Protocol for discrimination for automotive
paints by SEM-EDAX, Journal of Forensic Sciences, 35, 5, 1990
- Zieba-Palus et al, Mikrochim. Acta, 14, 1997.

I've taken same samples of automotive paints (such as Fiat, Peugeot,
Renault, Ford, Chevrolet, Volkswagen) corresponding to the years 1994-99.
I've found many difficulties while preparing them in:
- separating the paint from the body work of the car;
- cutting the specimen;
- to stick specimen to the graphite specimen pedestal;

Though I've made the primary morphological and chemical analysis and I
haven't observed great differences between the diferent automotive paints
trades (while polyuretane paints and belayer metalizer paints already tested),
but I'd found great differences between layers and layers of the paints
specimen.
As I'm not quite satisfied with the results obtained to be used for
forensic purposes, I would greatly appreciate making contacts with specialist
fo this subject, so as to receive any comments on this subject.
Thank you in advance for your help.
Jorgelina Altamirano
CERIDE,
Guemes 3450
(3000) Santa Fe, Argentina
e-mail: csedax-at-arcride.edu.ar




From: Mary Mager :      mager-at-interchange.ubc.ca
Date: Tue, 08 Feb 2000 09:49:38 -0800
Subject: Re: Be X-ray peaks -I don't think so

Contents Retrieved from Microscopy Listserver Archives
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Dear Scott,
I've seen a Be Ka x-ray peak in a Link advertisement and we have generated
one in the Quartz XOne applications lab with a really good light element
deteector. You will find the Be Ka1 peak listed at 0.110 keV in most x-ray
tables.
At 10:52 AM 2/7/00 -0500, you wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

Mary Mager
Electron Microscopist
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From: Christoph Bauer :      cbauer-at-midway.uchicago.edu
Date: Tue, 08 Feb 2000 09:56:35 -0800
Subject: Desmosomes

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we are looking at desmosomes in mouse epidermis. My question: From
looking at sections we got the impression that demosomes are rather
uniform, round knob-like structures. As we did not do any serial
sections, we are not sure if this is true. Does anybody know of a
publication dealing with this?

Thanks for your help,

Christoph



Christoph Bauer Ph.D.
University of Chicago
Molecular Genentics and Cell Biology
5841 S. Maryland Ave/MC 1028
Chicago, Il 60637




From: Ingber, Bruce F. :      bingber-at-commserver.srrc.usda.gov
Date: Tue, 8 Feb 2000 11:56:46 -0600
Subject: Re: FEI/Philips CM12 SAD focus

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I received the following fax from Ms. Irene Piscopo, EM Consultant,
regarding electron diffraction procedures for the CM-12 or CM-120 TEM's from
FEI/Philips. I'll copy the information for those interested.

ELECTRON DIFFRACTION PROCEDURES

There are three methods for doing electron diffraction in the CM-12
involving two different techniques: Method I is an aperture limiting method
while Methods II and III are probe limiting. In all cases, the sample must
be in the eucentric position and focused.

METHOD I: (area } 1 m) SAD

1. Select the SA mode (the diffraction aperture and image are focused in the
same plane).
2. Insert and center the SA aperture around the area of interest.*
3. Select D.
4. Remove the objective aperture.
5. Overfocus the second condenser to sharpen the pattern (i.e. parallel
illumination conditions).
6. Focus the pattern with the focus control which is now changing the
diffraction lens).
7. Change the size of the pattern (i.e. camera length) by changing the
magnification.

Size of the SA aperture
* Size of area selected =
----------------------------------------------------------------------------
------------
Objective lens magnification in D aperture
plane (~ 30x**)

** Actual magnification is focal length dependent.

METHOD II: (areas down to 40 nm with w filament; 20 nm with LaB6)

1. Locate an area of interest.
2. Remove the objective aperture.
3. Using Condensers 1 and 2, bring the beam down to a size slightly smaller
than the area of the area from which the diffraction pattern is to be
obtained.
4. Go to D and select the desired CL with the magnification control.
5. Focus the pattern with the focus control. Note: While it's sometimes
difficult to determine exact focus, it's often easier to insert an objective
aperture and focus the edge of the aperture to determine diffraction focus.
6. The size of the discs within the pattern can be decreased by decreasing
the size of the condenser aperture. Be sure the aperture is well-centered.

METHOD III: NANOPROBE and/or STEM (to 2 nm)

For diffraction and/or chemical information in the TEM mode from
regions of the sample less than 40 nm in diameter requires operating the
instrument in either the NANOPROBE or STEM modes.

NANOPROBE:

1. Depress NANOPROBE.
2. Follow instructions in Method II.

STEM:

1. Obtain a focused STEM image using the smallest C2 aperture.
2. Stop the scan.
3. Lower the main screen (CBED pattern)
4. Slightly larger probes and more parallel beam conditions can be obtained
using UUD and focusing with INTENSITY (C2 lens).

NOTES ON ELECTRON DIFFRACTION

1. The sample must always be eucentric and focused.
2. In METHOD I, the area from which the diffraction pattern is obtained is
determined by the selected area - (diffraction) aperture, when in SA mode.
To sharpen the ED pattern, one overfocuses the second condenser (C2)
3. In METHODS II and III, the area from which the diffraction pattern is
selected depends on the size of the beam. The size of the beam can be
altered by changing condenser lenses C1 and C2 (i.e. spot size and
intensity. To sharpen the ED pattern , one reduces the size of the C2
aperture.
4. Spherical aberration limits METHOD I to ~1 m.
5. In METHODS II and III, spherical aberration of the objective lens is not
the limiting factor, the minimum probe size is.
6. For identifying an ED pattern, at least three rings are required.
7. Be sure the sample and the standard are in the eucentric position.
Changes in the objective current will cause variations in CL.


Bruce F. Ingber, Biologist
USDA-ARS, SRRC
1100 Robert E. Lee Blvd.
New Orleans, LA 70124

(504) 286-4270 phone
(504) 286-4419 fax
bingber-at-nola.srrc.usda.gov






From: Walck. Scott D. :      walck-at-ppg.com
Date: Tue, 8 Feb 2000 12:57:15 -0500
Subject: RE: Be X-ray peaks -I don't think so

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I'm wrong. I guess I should have looked up at the X-ray periodic table
above my head and saw the 0.108 keV value for Be. The reason that I was
wrong was that I did not consider the solid state aspects. I am still right
about not having a 2s1/2 to a 1s1/2 transition and that an X-ray carries
away 1h-bar of angular momentum. What I didn't do was think about the band
structure of a solid. If you have the solid, the 2p bands of Be most be
spilling into or be the conduction band for the electrons. That must be the
source for the electronic transition. I guess I should have engaged brain
before typing.
My most humble apologies.
-Scott

Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)


"The opinions expressed are those of Scott D. Walck and not of PPG
Industries, Inc. nor of any PPG-associated companies."
--

} -----Original Message-----
} From: microprobe [mailto:chris.salter-at-materials.oxford.ac.uk]
} Sent: Tuesday, February 08, 2000 7:10 AM
} To: Walck. Scott D.
} Subject: Re: Be X-ray peaks -I don't think so
}
}
} Dear Walck,
}
} You may not, in theory, be able to produce Be X-rays, but we
} can detect them on our JEOL JXA 8800, they correspond to the position
} in the tables for Be Ka!, and the target is pure Be.
}
} On Mon, 7 Feb 2000 10:52:29 -0500 "Walck. Scott D." {walck-at-ppg.com}
} wrote:
}
} -----Original Message-----
} From: Ekstrom, Harry [mailto:harry.ekstrom-at-honeywell.com]
} Sent: Monday, February 07, 2000 7:40 PM
} To: 'Walck. Scott D.'
} Subject: RE: Be X-ray peaks -I don't think so
}
}
} I don't agree Scott. An L or M line for any element would
} not be located in
} the area
} of Be Ka. And the peak I normally get there with an ultra thin window
} is a well defined peak with great resolution.
} Please re evaluate your theory.
}
} Harry Ekstrom
} } }
} --------------------------------------------------------------
} ----------
} } 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
} }
}
} --------------------------------------------------------------
} ---------.
} }
} }
} } When a characteristic X-ray is given off from an atom, it
} carries 1 h-bar
} } (Planck's constant divided by 2 pi) of angular momentum.
} The electronic
} } transition that occurred for the X-ray to come off must
} conserve angular
} } momentum of the system. Therefore, a transition from a
} 2s1/2 to a 1s1/2
} } state is forbidden by selection rules. You can't produce a
} Be X-ray.
} } -Scott
} }
} } Scott D. Walck, Ph.D.
} } PPG Industries, Inc.
} } Glass Technology Center
} } Guys Run Rd. (packages)
} } P. O. Box 11472 (letters)
} } Pittsburgh, PA 15238-0472
} }
} } Walck-at-PPG.com
} }
} } (412) 820-8651 (office)
} } (412) 820-8161 (fax)
} }
} }
} }
}
} -------------------
} microprobe
} chris.salter-at-materials.ox.ac.uk
}
} * This e-mail message was sent with Execmail V5.0.x *
}




From: Steve Chapman :      PROTRAIN-at-CompuServe.COM
Date: Tue, 8 Feb 2000 15:31:08 -0500
Subject: Diffraction & Coherence

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Hi

It has been very interesting reading about diffraction and electron beams
but it is quite clear that we often give the instrument credit, "the
instrument does it automatically", when credit is not due. Almost every
"automatic" is a designers estimate or better still a calculation, it is a
setting!

After 36 years with TEM I have to say that the only area that the
instrument could set in the SA mode could be the approximate position of
the first image plane. We normally move to the SA mode to free up the
diffraction lens (1st intermediate) to enable it to be focussed on thefirst
image plane and the diffraction (intermediate) aperture, independent of the
magnification system. We would then focus the image inside the aperture
with the objective lens to ensure that the selected area IS the area in the
diffraction pattern. Without this procedure electron rotation and
misalignments could give you a diffraction pattern from an area not in the
centre of the screen! Users are correct in that if they set the eucentric
point or better still, adjust the specimen height to focus at the same
objective lens current value, that done the first image plane will be at
the same point within the diffraction (intermediate) lens.

So how could the Philips work? I would guess they set the diffraction lens
at approximately the first image plane. Then as they do not use it within
their lens scheme for the SA magnifications it is fixed at this focus, good
enough I think?

A second point, parallel beams are very important if you are chasing the
best image quality, biology or materials, most times we need high
coherence. There are misunderstandings on how to obtain a parallel beam or
high coherence, setting the final condenser underfocus is incorrect. The
procedure for high coherence would be to use the smallest spot size you can
tolerate (this probably means you must up the emission current). Once in
this condition overfocus the final condenser (clockwise from crossover) the
spot, what ever size it is now, becomes your virtual source. The further
overfocus you go the greater the beam coherence . You will deduce the
smaller the condenser aperture the sharper the spot and the smaller the
spot the greater the coherence for a given degree of overfocus.

Work with a design team and they expect everyone to overfocus, they expect
everyone to use small condenser apertures and they expect everyone to use
small spot sizes. They do not expect everyone to use too low an emission
current (almost everyone does, we have talked before about filament life
being the most important feature of many laboratories) because this makes
the task too difficult!


Steve Chapman
Senior Consultant
Protrain
Electron microscopy courses world wide
http://www.emcourses.com
Tel & Fax 44 (0) 1280 814774




From: Schibler, Matthew :      MSchibler-at-mednet.ucla.edu
Date: Tue, 8 Feb 2000 12:42:13 -0800
Subject: RE: HeNe laser

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} Dear all

} I am planning to replace my Kr/Ar laser on a Zeiss 310 CSLM with two
} separate lasers: 488nm Ar ion and HeNe 543nm. Does anyone have
} experience of using a HeNe 543? I need this or something similar to
} excite Rhodamine for food structure applications. I'd appreciate any
} advice.

} The reason I'm replacing the mixed gas laser is that has
} averaged 25% downtime over the past three years.

} Mark

} Mark Auty
} Dairy Products Research Centre
} Moorepark, Fermoy, Co. Cork, Ireland.
} mauty-at-moorepark.teagasc.ie
} tel 00353 2542447
} fax 00353 2542340


I have been using the Argon ion (488nm) and HeNe 543nm combination on
different Zeiss LSM310's and 410's for the past 9 years. The HeNe 543nm is
fairly low power, but it has been perfectly adequate for scanning the
fluorochromes I have needed to see including TRITC, Texas Red, Cy3,
propidium iodide, and Sirius Red. I have never had a problem with the laser
going down either, but one would expect that with a HeNe laser.

Slinte,

Matthew J. Schibler Ph.D.
UCLA Brain Research Institute
1524A Gonda (Goldschmied) Center
for Neuroscience and Genetics
Los Angeles, CA 90095-1761

(310) 825-9783
FAX (310) 206-5855
E-mail: mschibler-at-mednet.ucla.edu




From: Kristen Lennon :      kalen-at-citrus.ucr.edu
Date: Tue, 08 Feb 2000 14:06:58 -0800
Subject: cryo chamber for LRWhite UV

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Hi All,
Does anyone have experience with the Pelco UVC2 or UVC1 Cryo Chamber for UV
curing of resins (especially LR White) or with something similar? We're
contemplating buying something like that for UV polymerization of LR White
at -20 C.
Thanks for your help,
Kristen
Kristen A. Lennon
Cell, Molecular & Developmental Biology Group
Department of Botany & Plant Sciences
University of California
Riverside, CA 92521
kalen-at-citrus.ucr.edu




From: Jim McGee :      mcgee-at-geol.sc.edu
Date: Tue, 08 Feb 2000 17:09:09 -0500
Subject: Re: Be X-ray peaks -I don't think so

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Scott,
I'm not sure what brought this up, but my reaction is "thems fightin'
words". I do not currently have a wavelength reflecting crystal to measure
that low in the periodic table, but I did on another instrument. I distinctly

recall seeing a pretty hefty peak at the Be K-alpha position when I focused
the beam down on a piece of pure Be metal. I guess rules are made to be
broken. Are your calculations correct? Or am I misunderstanding something
here?

Jim
--
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
James J. McGee (email: jmcgee-at-sc.edu)
Department of Geological Sciences
University of South Carolina
Columbia, SC 29208

Tel: 803-777-6300 Fax: 803-777

"Walck. Scott D." 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
} -----------------------------------------------------------------------.
}
} When a characteristic X-ray is given off from an atom, it carries 1 h-bar
} (Planck's constant divided by 2 pi) of angular momentum. The electronic
} transition that occurred for the X-ray to come off must conserve angular
} momentum of the system. Therefore, a transition from a 2s1/2 to a 1s1/2
} state is forbidden by selection rules. You can't produce a Be X-ray.
} -Scott
}
} Scott D. Walck, Ph.D.
} PPG Industries, Inc.
} Glass Technology Center
} Guys Run Rd. (packages)
} P. O. Box 11472 (letters)
} Pittsburgh, PA 15238-0472
}
} Walck-at-PPG.com
}
} (412) 820-8651 (office)
} (412) 820-8161 (fax)





From: Rosemary Walsh :      rw9-at-psu.edu
Date: Tue, 8 Feb 2000 16:24:35 -0600
Subject: Size determination of overlapping particles

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Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: Rosemary Walsh :      rw9-at-psu.edu
Date: Tue, 8 Feb 2000 16:24:35 -0600
Subject: Size determination of overlapping particles

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Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: Rosemary Walsh :      rw9-at-psu.edu
Date: Tue, 8 Feb 2000 16:24:35 -0600
Subject: Size determination of overlapping particles

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Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: Rosemary Walsh :      rw9-at-psu.edu
Date: Tue, 8 Feb 2000 16:24:35 -0600
Subject: Size determination of overlapping particles

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Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: Rosemary Walsh :      rw9-at-psu.edu
Date: Tue, 8 Feb 2000 16:24:35 -0600
Subject: Size determination of overlapping particles

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Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: Rosemary Walsh :      rw9-at-psu.edu
Date: Tue, 8 Feb 2000 16:24:35 -0600
Subject: Size determination of overlapping particles

Contents Retrieved from Microscopy Listserver Archives
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Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: Rosemary Walsh :      rw9-at-psu.edu
Date: Tue, 8 Feb 2000 16:24:35 -0600
Subject: Size determination of overlapping particles

Contents Retrieved from Microscopy Listserver Archives
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Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: Rosemary Walsh :      rw9-at-psu.edu
Date: Tue, 8 Feb 2000 16:24:35 -0600
Subject: Size determination of overlapping particles

Contents Retrieved from Microscopy Listserver Archives
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Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: Rosemary Walsh :      rw9-at-psu.edu
Date: Tue, 8 Feb 2000 16:24:35 -0600
Subject: Size determination of overlapping particles

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: Rosemary Walsh :      rw9-at-psu.edu
Date: Tue, 8 Feb 2000 16:24:35 -0600
Subject: Size determination of overlapping particles

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: Rosemary Walsh :      rw9-at-psu.edu
Date: Tue, 8 Feb 2000 16:24:35 -0600
Subject: Size determination of overlapping particles

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: Rosemary Walsh :      rw9-at-psu.edu
Date: Tue, 8 Feb 2000 16:24:35 -0600
Subject: Size determination of overlapping particles

Contents Retrieved from Microscopy Listserver Archives
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Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: Dr Eric Lachowski :      che136-at-abdn.ac.uk
Date: Tue, 8 Feb 2000 16:24:58 -0600
Subject: Be X-ray and filament drift

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Dear Listers,
I have responses to two of today's postings, but they are
quite unrelated.
1. I did not see the original question about Be x-rays,
only Scott Walk's reply, so I don't know what info was
being sought. We used a Be-containing mineral to evaluate
microprobes before buying. I won't embarrass those who
failed, but only one manufacturer succeeded. The reason
the others failed is probably that in compounds the x-ray
energy is subject to largish chemical shifts relative to
pure Be because there is no shielding by outer electrons
and you need to search on either side of the tabulated
value of about 0.1keV. So there IS a Be x-ray and a
reasonable WDS spectrometer should detect it.

2. Filament drift. Fred Schamber's reply is right most of
the time, but I did once have a batch of 6 JEOL-type
filaments made by an EM supply house where 2 of them
drifted until the tips were right at the edge of the hole
and stayed there even when the filament was cooled down
again. This was the only time in about 15 years of using
JEOL microscopes.

Eric
----------------------
Dr Eric Lachowski
Department of Chemistry
University of Aberdeen
Meston Walk
Old Aberdeen AB24 3UE
Scotland
+44 (0)1224 272934 fax 272921
e.lachowski-at-abdn.ac.uk






From: Dean Armytage :      hillstream-at-hypermax.net.au
Date: Tue, 8 Feb 2000 16:27:01 -0600
Subject: Live Blood Staining

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Hi List I am using live blood staining light microscopy, is there
anyone who has tried or is using this technique. I would like to swap
notes. Dean Armytage PhD Hillstream Health Centre Australia






From: Ekstrom, Harry :      harry.ekstrom-at-honeywell.com
Date: Tue, 8 Feb 2000 18:55:31 -0700
Subject: Re: Be X-ray peaks -I don't think so

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I'm getting more curious as I read on here. Speaking strictly energy now
and leaving out wavelength, the characteristic energy of Be Ka is about .11
KeV like Mary stated. I've had an occasional peak show up there before and
thought it may have been Be. Now I understand that it may have been "noise"
all along. Would "noise" be a pretty defined and relatively well resolved
peak at that energy level using an ultra thin window? Depending on the low
energy cutoff setting, one might truly have Be and never detect it. Would
lowering the KV reduce the noise artifact?

I guess I'll get out my Be planchet and try a few things.

Harry

-----Original Message-----
} From: Jim McGee [mailto:mcgee-at-geol.sc.edu]
Sent: Tuesday, February 08, 2000 3:09 PM
To: 'Microscopy'


Scott,
I'm not sure what brought this up, but my reaction is "thems fightin'
words". I do not currently have a wavelength reflecting crystal to measure
that low in the periodic table, but I did on another instrument. I
distinctly

recall seeing a pretty hefty peak at the Be K-alpha position when I focused
the beam down on a piece of pure Be metal. I guess rules are made to be
broken. Are your calculations correct? Or am I misunderstanding something
here?

Jim
--
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
James J. McGee (email: jmcgee-at-sc.edu)
Department of Geological Sciences
University of South Carolina
Columbia, SC 29208

Tel: 803-777-6300 Fax: 803-777

"Walck. Scott D." 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
} -----------------------------------------------------------------------.
}
} When a characteristic X-ray is given off from an atom, it carries 1 h-bar
} (Planck's constant divided by 2 pi) of angular momentum. The electronic
} transition that occurred for the X-ray to come off must conserve angular
} momentum of the system. Therefore, a transition from a 2s1/2 to a 1s1/2
} state is forbidden by selection rules. You can't produce a Be X-ray.
} -Scott
}
} Scott D. Walck, Ph.D.
} PPG Industries, Inc.
} Glass Technology Center
} Guys Run Rd. (packages)
} P. O. Box 11472 (letters)
} Pittsburgh, PA 15238-0472
}
} Walck-at-PPG.com
}
} (412) 820-8651 (office)
} (412) 820-8161 (fax)





From: Hong Yi :      hyi-at-emory.edu
Date: Tue, 8 Feb 2000 21:26:04 -0500 (EST)
Subject: Re: New Gold Enhancement Reagents

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Dear Michael:

My personal experience with silver enhancement dates back to when Janssen
Life Sciences first introduced the IntenSE M silver enhancement kit in
late 80's (this product was later taken over by Amersham). At the time, I
felt the IntenSE M was easy to use, but its fast reaction made it hard to
control the particle growth. Just like you, the desire for a better
results has kept me searching and testing whenever a new protocol or
product became available.

Danscher's method gives wonderful intensification, but its low pH
sometimes damages the ultrastructure when used for the pre-embedding
immunogold labeling. Burry's method and the Nanoprobes HQ silver made
progress with pH, however they inherited Danscher's high viscosity and
light sensitivity, which limit their penetration in pre-embedding silver
enhancement and render it more difficult to handle from a practical
standpoint. I have also tested the Nanoprobes GoldEnhance kit for
pre-embedding immunogold labeling recently. At the LM level the results
were decent. I have not spent a lot of time evaluating its performance
at the EM-level. So far, the particle size homogeneity is not as good as
what I've found with Danscher's method. But I will refrain from further
judgment until more work has been done.

In my opinion, the first breakthrough in intensifying gold particles since
Danscher's method was realized when Aurion, a Dutch company, first
introduced the R-gent SE-EM silver enhancement kit last June. As you
wished for in your post, it has a near-neutral pH, low viscosity, and it
is light insensitive. Moreover, it gives a great enhancement efficiency
and even particle size and shape. Background remains minimal even after
two hours enhancement on the bench. Morphology preservation after
enhancement is superior to any other enhancement reagent I have used. I
have been testing this kit profusely for both post- and pre-embedding
immunogold labeling on various types of samples (including cell cultures
and vibratome sections), with many different primary and secondary
antibodies, and am very pleased with its performance. I use 0.5% OsO4 for
20 min and have never had any problem with silver disappearing. If you
like, I would be very happy to e-mail some images to you so you can
evaluate them yourself.

Hong
=================
Hong Yi
Emory University
Department of Neurology
6215 Woodruff Memorial Research Building
1639 Pierce Dr.
Atlanta, GA 30341
Phone: (404) 727-8692
Fax: (404) 727-3157
Email: hyi-at-emory.edu

On Thu, 3 Feb 2000, Michael Plociniak wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
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} -----------------------------------------------------------------------.
}
}
} Dear Colleagues,
} Has anyone used gold enhancement reagents from Nanoprobes, Inc.
} (Stonybrook, NY - USA) as an alternative to silver enhancement for
} enlarging 1 nm immunogold probes?
}
} Advertisements state that gold enhancement has lower background and is
} compatible with osmium. Can anyone verify this from personal experience?
}
} In addition to these advantages, I am hoping that milder pH conditions will
} be less damaging to ultrastructure in cultured neurons (using a
} preembedment protocol).
}
} Thank you,
} Michael
}







From: Radostin Danev :      rado-at-nips.ac.jp
Date: Wed, 9 Feb 2000 12:01:47 +0900
Subject: Finding the diffraction plane

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hi everybody,

Here is the way I found yesterday for finding the diffraction plane. By
diffraction plane I mean the plane at which diffraction pattern is formed
(crossover, Fourier of the object wave) not the theoretical BFP of the
objective.

First perform standard alignment for brightfield mode. Then go to 10 000X.
There are several ways to perform:

1. Fixed illumination and specimen position (i.e. OL current)

a) Adjust the desired brightness with C2. Adjust the desired OL current and
specimen position.
b) Insert OL aperture and move it so that the edge of the aperture to cross
through the middle of the screen. If the aperture is exactly at the diff.
plane then you'll not be able to see its edge - just the whole beam will
fade uniformly ... so you have the diff. plane.
c) If you see aperture edge then it is not at diff. plane. Now two ways:
- change the aperture z-position (not recommended ... but if it is very far
..)
- If your microscope has condenser minilens you can try tweaking both C2 and
CML in order to keep the intensity same and just make the aperture edge
disappear (i.e. the whole screen uniformly darkened).

2. Fixed specimen (i.e. OL current).

a) Focus the specimen
b) same as 1b
c) Change C2 excitement in order to make the edge disappear

3. Fixed illumination

a) Set desired C2 excitement
b) same as 1b
c) Change the objective lens current until the aperture edge disappears.
Then move specimen in z-direction until it is focused.


In all these methods after adjusting the aperture at diff. plane you can try
tweaking C2 or OL ... you will see that the aperture is not at diff. plane
anymore, The edge will appear on one or the other side (i.e. mirrored)
depending on the direction of the lens current change.
Another thing - after adjusting the aperture at diff. plane you can check if
beam shift tilts the beam (by tilting here I mean - if the specimen is
illuminated with plane wave then shifting the beam should not tilt the wave
front). Shift the beam and if the brightness changes (when the aperture edge
partially covers the zero diff. spot) then the beam is tilting.
I wish there was a IL1 tweaking knob allowing for change of the IL1 current
while in brightfield mode. This wold make the things much more easier.

About the methods for making the illumination at the specimen parallel. They
will work only if:
- The OL aperture is positioned by the manufacturer exactly at the
theoretical BFP (I think that in most of the TEMs it is not)
- The OL current should be set to zero deviation (i.e. at the value for
which the BFP position was calculated)
- The intermediate lens system should be properly calibrated by the
manufacturer so that if both of the above conditions are satisfied and the
specimen is at front focal plane of the OL then it to be in focus. In other
words the theoretical BFP of the objective to coincide with the theoretical
front focal plane of the IL1.

I don't think parallel illumination is so very important ... using spherical
wave will give the same results but just the diff. planes will be shifted.

All of the above is just my opinion. I could be mistaking somewhere ...

Best wishes,

Rado

---------------------------------------------------------------------
Radostin Danev
Laboratory of Ultrastructure Research
National Institute for Physiological Sciences
Myodaiji-cho, Okazaki 444-8585, JAPAN
e-mail: rado-at-nips.ac.jp
---------------------------------------------------------------------





From: Gordon Couger :      gcouger-at-couger.com
Date: Tue, 8 Feb 2000 21:31:38 -0600
Subject: Re: automotive paints

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html



----- Original Message -----
} From: "Dennis Ward" {DCWard-at-concentric.net}
}
} Jorgelina
} SEM/EDS//EPMA is only one class of analytical methods that the
Forensic
} community applies to paint analysis for comparison and association with
} original source. Although used routinely, probe methods are not used
} exclusively. The organic components in paints are generally more
} discriminating.
} Sample preparation usually entails some form of embedment and either
} microtomy or polishing in order to reveal internal structure.
} Removal of paint from an auto body is tricky, and requires
practice.The
} manufacturers have designed their paint systems to prevent removal!
} I would be glad to provide you with additional resources.

I have never do this but this is what i would try first.

One thing I would try on would be removing the metal from the paint. A
weak elecrolite solution and a low DC voltage connected to reverse
electoplate the metal away sould get rid of almost all of it and the last
bit could be removed with acid or evaporated as the cathode in a vacum
chamber.

Once you get the metal down to a bunch of small islands you might be able
to let it rust free of the paint.

I have no idea what this would do to the paint but most paint films I have
delt with are pretty tough.

Gordon

Gordon Couger gcouger-at-couger.com

Stillwater, OK www.couger.com/gcouger
405 624-2855 GMT -6:00






From: Massimo Catalano :      massimo.catalano-at-ime.le.cnr.it
Date: Wed, 09 Feb 2000 09:43:03 +0100
Subject: PIPS and milling damage

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Listservers,

we are massively working on analytical and structural TEM characterization
of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots.

Specimen preparation is of course a crucial point of our work.

We have just switched to PIPS (we used to make our samples using the Gatan
Duo Mill), and we are getting controversial results about the damage
introduced by the milling procedure.

We are perfectly aware of all the differences between the two instruments,
especially the absence of cooling stage and the higher beam current.

Has anyone performed a systematic and careful analysis to try to evaluate
the milling damage on similar materials systems. We are willing to start a
systematic work to assess this issue, but would like to know if anyone has
already done anything on this topic. Also, any collaboration will be more
than welcome.

Thanks.

Massimo




Dr. Massimo Catalano
CNR-IME
Campus Universitario
Via Arnesano
73100 Lecce - ITALY
tel: + 39 0832 322362
fax: + 39 0832 325299
email: massimo.catalano-at-ime.le.cnr.it
http://www.ime.le.cnr.it
http://www.ime.le.cnr.it/sime/sime.htm






From: flaitz-at-us.ibm.com
Date: Wed, 9 Feb 2000 07:48:09 -0500
Subject: PIPS and milling damage

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html



Dear Massimo,

in our lab we have prepared a number of GaAs, AlGaAs, InGaAs devices.
Mechanical thinning is by the tripod wedge technique, but typically we end
up with more damage than we find for Si samples so there is a need to leave
samples thicker and use more extensive ion milling for final thinning.

We also have a PIPS and I have found its capabilities essential for
achieving good thin samples in these materials. I found more success by
using low angles and low KeV for long times. Typically I would be using
5-6 degrees and 4-4.5 KeV initially, with 3-4 degrees and 3-3.5 KeV for the
final ion polishing. On samples 2-3 microns thick, this would amount to
2-3 hours total milling time. I did not observe any artefacts introduced
with milling under these conditions, and the junction structures and
inherent defects were clearly observed.

The conditions above should work with any ion miller capable of low angle
milling, but our experience is only with the PIPS. If you use the wedge
technique for mechanical thinning, I would recommend that you use Mo grids
to mount your specimens as the extended milling times at low angles will
really chew up a Cu grid. Also, Mo grids are considerably thinner,
allowing angles as low as 3-4 degrees on the grid hole side for a sample
positioned in the middle of a 1 mm grid hole.

If you would like, I can send you some typical images of devices we have
prepared, including images showing the removal of the mechanical polishing
damage.


Philip L. Flaitz
IBM Analytical Services, Hopewell Junction, NY
http://www.chips.ibm.com/services/asg
Ph.......(914) 892-3094, FAX -2003
flaitz-at-us.ibm.com



Massimo Catalano {massimo.catalano-at-ime.le.cnr.it} on 02/09/2000 03:43:03 AM

To: Microscopy-at-sparc5.Microscopy.Com
cc:


Dear Listservers,

we are massively working on analytical and structural TEM characterization
of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots.

Specimen preparation is of course a crucial point of our work.

We have just switched to PIPS (we used to make our samples using the Gatan
Duo Mill), and we are getting controversial results about the damage
introduced by the milling procedure.

We are perfectly aware of all the differences between the two instruments,
especially the absence of cooling stage and the higher beam current.

Has anyone performed a systematic and careful analysis to try to evaluate
the milling damage on similar materials systems. We are willing to start a
systematic work to assess this issue, but would like to know if anyone has
already done anything on this topic. Also, any collaboration will be more
than welcome.

Thanks.

Massimo




Dr. Massimo Catalano
CNR-IME
Campus Universitario
Via Arnesano
73100 Lecce - ITALY
tel: + 39 0832 322362
fax: + 39 0832 325299
email: massimo.catalano-at-ime.le.cnr.it
http://www.ime.le.cnr.it
http://www.ime.le.cnr.it/sime/sime.htm










From: LAB. DE MICROSCOPIA ELECTRONICA - FI - UNER :      microsc-at-fi.uner.edu.ar
Date: Wed, 9 Feb 2000 13:10:00 +0000
Subject: looking for ETEC Company

Contents Retrieved from Microscopy Listserver Archives
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Hello .....
we have an old ETEC Omniscan SEM, and we are searching for ETEC company or any
people that can supply manuals ....

any help is welcome

best regards


===================================================
Fernando D. Balducci
Laboratorio de Microscopia Electrnica
Facultad de Ingeniera - Bioingenieria
Universidad Nacional de Entre Rios
Argentina
e-mail: microsc-at-fi.uner.edu.ar
tel: 54 43 975100
fax: 54 43 975077


===================================================




From: Corazon Bucana :      bucana-at-audumla.mdacc.tmc.edu
Date: Wed, 09 Feb 2000 08:11:32 -0600
Subject: autofluorescence quenching

Contents Retrieved from Microscopy Listserver Archives
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I am working with archival specimens of formalin-fixed paraffin embedded
tissues and I am getting tremendous autofluorescence. I looked at tissue
sections after the sections have been de-waxed using either FITC or
rhodamine filter sets and red cells and connective tissues are brightly
fluorescent. Is there a way of suppressing the autofluorescence and still
retain reactivity of tissue to antibodies? I would appreciate any comments
or suggestions.

*******************************************************
Corazon D. Bucana, Ph.D.
Department of Cancer Biology
U.T. M.D. Anderson Cancer Center
1515 Holcombe Blvd. Box 173
Houston, Texas 77030
Phone: (713) 792-8106
FAX: (713) 792-8747
Email:bucana-at-audumla.mdacc.tmc.edu
FAX: (713) 792-8747





From: Jim McGee :      mcgee-at-geol.sc.edu
Date: Wed, 09 Feb 2000 10:26:27 -0500
Subject: Re: Be X-ray peaks -I don't think so

Contents Retrieved from Microscopy Listserver Archives
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Harry,

I don't have a UTW detector here, so I'll leave this to the EDS experts. But,
on the JEOL 8800/8900 I used to run we had a thin window detector that could see
B (not Be) if the low end noise peak (which is huge) was properly
discriminated. Get out your C planchett while you are getting the Be one, and
see if you get the noise peak at Be. Far better to use wavelength than EDS down
at that end.

Jim McGee
--
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
James J. McGee (email: jmcgee-at-sc.edu)
Department of Geological Sciences
University of South Carolina
Columbia, SC 29208

Tel: 803-777-6300 Fax: 803-777-6610

"Ekstrom, Harry" 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'm getting more curious as I read on here. Speaking strictly energy now
} and leaving out wavelength, the characteristic energy of Be Ka is about .11
} KeV like Mary stated. I've had an occasional peak show up there before and
} thought it may have been Be. Now I understand that it may have been "noise"
} all along. Would "noise" be a pretty defined and relatively well resolved
} peak at that energy level using an ultra thin window? Depending on the low
} energy cutoff setting, one might truly have Be and never detect it. Would
} lowering the KV reduce the noise artifact?
}
} I guess I'll get out my Be planchet and try a few things.
}
} Harry
}
} -----Original Message-----
} } From: Jim McGee [mailto:mcgee-at-geol.sc.edu]
} Sent: Tuesday, February 08, 2000 3:09 PM
} To: 'Microscopy'
} Subject: Re: Be X-ray peaks -I don't think so
}
} ------------------------------------------------------------------------
} 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
} -----------------------------------------------------------------------.
}
} Scott,
} I'm not sure what brought this up, but my reaction is "thems fightin'
} words". I do not currently have a wavelength reflecting crystal to measure
} that low in the periodic table, but I did on another instrument. I
} distinctly
}
} recall seeing a pretty hefty peak at the Be K-alpha position when I focused
} the beam down on a piece of pure Be metal. I guess rules are made to be
} broken. Are your calculations correct? Or am I misunderstanding something
} here?
}
} Jim
} --
} ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
} James J. McGee (email: jmcgee-at-sc.edu)
} Department of Geological Sciences
} University of South Carolina
} Columbia, SC 29208
}
} Tel: 803-777-6300 Fax: 803-777
}
} "Walck. Scott D." 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
} } -----------------------------------------------------------------------.
} }
} } When a characteristic X-ray is given off from an atom, it carries 1 h-bar
} } (Planck's constant divided by 2 pi) of angular momentum. The electronic
} } transition that occurred for the X-ray to come off must conserve angular
} } momentum of the system. Therefore, a transition from a 2s1/2 to a 1s1/2
} } state is forbidden by selection rules. You can't produce a Be X-ray.
} } -Scott
} }
} } Scott D. Walck, Ph.D.
} } PPG Industries, Inc.
} } Glass Technology Center
} } Guys Run Rd. (packages)
} } P. O. Box 11472 (letters)
} } Pittsburgh, PA 15238-0472
} }
} } Walck-at-PPG.com
} }
} } (412) 820-8651 (office)
} } (412) 820-8161 (fax)

--
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
James J. McGee (email: jmcgee-at-sc.edu)
Department of Geological Sciences
University of South Carolina
Columbia, SC 29208

Tel: 803-777-6300 Fax: 803-777-6610






From: ipaul-at-MtRoyal.AB.CA
Date: Wed, 09 Feb 2000 09:26:15 -0700
Subject: Sarcomere cross-sections

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


I am looking for TEM images that show cross-sectional views of skeletal muscle
sarcomeres during the *contracted state*. In particular, I am interested in
seeing the spacing of the thin filaments when they **overlap each other** (i.e.,
when those from one side of the sarcomere overlap those from the other side of
the sarcomere in the contracted state). In addition, I am interested in seeing
the distribution of the thin filaments as they pass through the M line. It
would be greatly appreciated if anyone could tell me if they know of any
research papers or review articles that contain such TEM images.

Thank you!

Izak Paul
Biological Sciences
Mount Royal College






From: David Knecht :      knecht-at-uconn.edu
Date: Wed, 9 Feb 2000 13:03:36 -0500
Subject: Paultek camera

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Does anyone know if Paultek Imaging still exists and a phone number for
them (or Email)? Thanks-Dave


************************************************************

Dr. David Knecht
Department of Molecular and Cell Biology
University of Connecticut
75 N. Eagleville Rd. U-125
Storrs, CT 06269
Knecht-at-uconnvm.uconn.edu
860-486-2200 860-486-4331 (fax)
home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
************************************************************






From: Roger Moretz :      rcmoretz-at-excite.com
Date: Wed, 9 Feb 2000 10:06:34 -0800 (PST)
Subject: Re: Desmosomes

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Christoph:
I don't have any references at hand, but there is a whole literature on
desmosomes. A search on Medline (or PubMed on the internet) should provide
you with a good list of EM related publications on desmosomes--probably more
than you ever wanted to know! As an aside, I think there may be some books
that cover this as well, but since it is not an area of personal expertise
or current research interest, I am afraid they have all gone from my memory
banks.

Roger


On Tue, 08 Feb 2000 09:56:35 -0800, Christoph Bauer 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 are looking at desmosomes in mouse epidermis. My question: From
} looking at sections we got the impression that demosomes are rather
} uniform, round knob-like structures. As we did not do any serial
} sections, we are not sure if this is true. Does anybody know of a
} publication dealing with this?
}
} Thanks for your help,
}
} Christoph
}
}
}
} Christoph Bauer Ph.D.
} University of Chicago
} Molecular Genentics and Cell Biology
} 5841 S. Maryland Ave/MC 1028
} Chicago, Il 60637
}


Roger C Moretz, Ph.D.





_______________________________________________________
Get 100% FREE Internet Access powered by Excite
Visit http://freeworld.excite.com





From: Mary Gail Engle :      mgengle-at-pop.uky.edu
Date: Wed, 09 Feb 2000 10:06:35 -0800
Subject: Re: Desmosomes

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Christoph,
I have examined alot of mouse and pig skin at the TEM level and the
desmosomes and hemidesmosomes appear rather typical, that is, small, long
bridge-like structures between the cells.

Mary Gail Engle
Electron Microscopy & Imaging Facility
University of Kentucky

At 09:56 AM 2/8/00 -0800, Christoph Bauer wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America





From: Fagerland,Jane :      jane.a.fagerland-at-abbott.com
Date: Wed, 9 Feb 2000 16:45:16 -0600
Subject: Summer internships

Contents Retrieved from Microscopy Listserver Archives
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The Department of Microscopy and Microanalysis at Abbott Laboratories has two
summer internships available for 8-12 weeks this summer. One position is in
Biological Microscopy, and one is in Materials Analysis and Microscopy.
We're looking for students who are considering a career in microscopy,
especially students interested in the pharmaceutical industry.

Abbott Laboratories is a diversified healthcare company that produces
pharmaceuticals, diagnostic devices, and hospital products. The Microscopy
department analyzes samples related to all these functions. We use polarized
light, fluorescence, TEM, SEM, EDXS, confocal microscopy, and flow cytometry
to solve problems related to products, as well as to provide support for
pharmaceutical Discovery and Development basic research and drug safety.

Housing and a stipend are provided for the summer. We're located near Lake
Michigan and the Wisconsin state border, about an hour's drive or train-ride
north of Chicago. There's lots to see and do in the area, and there are
planned activities with other interns, as well. The Abbott Summer Internship
Program has been nationally recognized as one of the best in the country.

If you're interested, please send a resume to:

Jane A. Fagerland, Ph.D.
Abbott Laboratories
D-45M/AP31
200 Abbott Park Rd.
Abbott Park IL 60064-6202

You may also fax a resume to me at (847) 938-5027 or e-mail it to me at
jane.a.fagerland-at-abbott.com.

If you'd like further information, I can be reached by telephone at (847)
935-0104, or by e-mail. I prefer e-mail, but will be more than happy to talk
to discuss our projects and laboratory by telephone.




From: Walck. Scott D. :      walck-at-ppg.com
Date: Wed, 9 Feb 2000 20:31:58 -0500
Subject: RE: PIPS and milling damage

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


There is an easy way to determine the milling damage between the two
machines and the amount of ion milling damage (subjectively); compare them
with samples prepared using the small angle cleavage technique. Look at
John McCaffrey's paper on using the small angle cleavage technique in
Ultramicrotomy 38, (149) 1991. He shows the difference between high angle
milling, low angle milling and the small angle cleavage technique. Of
course, the small angle cleavage technique showed no ion milling damage. It
is perfect for these types of materials. If you don't need a site specific
technique, it is the way to go for semiconductors.

You should also look at his paper in the MRS TEM Sample Prep series III (vol
254) that also shows a comparison for a SiGe/Si layer structure. We have a
detailed pictorial outline with tips and tricks on how to do it in the MRS
TEM Sample Prep series IV (vol 480).

I know that you invested a lot in your ion mill, but you can do these
samples very cheaply. SouthBay Technology sells a Microcleave kit that gets
you started with the technique.



Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)


"The opinions expressed are those of Scott D. Walck and not of PPG
Industries, Inc. nor of any PPG-associated companies."
--




} -----Original Message-----
} From: Massimo Catalano [mailto:massimo.catalano-at-ime.le.cnr.it]
} Sent: Wednesday, February 09, 2000 3:43 AM
} To: Microscopy-at-sparc5.microscopy.com
} Subject: PIPS and milling damage
}
}
} --------------------------------------------------------------
} ----------
} 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 Listservers,
}
} we are massively working on analytical and structural TEM
} characterization
} of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots.
}
} Specimen preparation is of course a crucial point of our work.
}
} We have just switched to PIPS (we used to make our samples
} using the Gatan
} Duo Mill), and we are getting controversial results about the damage
} introduced by the milling procedure.
}
} We are perfectly aware of all the differences between the two
} instruments,
} especially the absence of cooling stage and the higher beam current.
}
} Has anyone performed a systematic and careful analysis to try
} to evaluate
} the milling damage on similar materials systems. We are
} willing to start a
} systematic work to assess this issue, but would like to know
} if anyone has
} already done anything on this topic. Also, any collaboration
} will be more
} than welcome.
}
} Thanks.
}
} Massimo
}
}
}
}
} Dr. Massimo Catalano
} CNR-IME
} Campus Universitario
} Via Arnesano
} 73100 Lecce - ITALY
} tel: + 39 0832 322362
} fax: + 39 0832 325299
} email: massimo.catalano-at-ime.le.cnr.it
} http://www.ime.le.cnr.it
} http://www.ime.le.cnr.it/sime/sime.htm
}
}
}




From: Barbara Foster :      mme-at-map.com
Date: Wed, 09 Feb 2000 20:34:34 -0500
Subject: Re: LM: Course reminder - DATES...

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hi again,

It has been brought to my attention that no dates were attached to this
course reminder:

March 10-12, 2000
Hyatt Regency
New Orleans, LA.

Best regards,
Barbara Foster
Consortium President
Microscopy/Microscopy Education ...Educating microscopists for greater
productivity.

125 Paridon Street Suite 102 Springfield, MA 01118
PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Visit our web site {http://www.MME-Microscopy.com/education}



At 07:57 PM 2/7/00 -0500, Barbara Foster wrote:
} ------------------------------------------------------------------------
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From: Barbara Foster :      mme-at-map.com
Date: Wed, 09 Feb 2000 20:39:10 -0500
Subject: Re: tacky wax

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Linda,

Suggest you contact Chuck Garber at Structure Probe: www.2spi.com
I'm sure that he has some sort of derivative of his tacky dots which would
be helpful


At 08:48 PM 8/27/70 +0000, Linda Chicoine wrote:
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From: Christina bennett :      chbennet-at-nmsu.edu
Date: Wed, 9 Feb 2000 21:58:18 -0700
Subject: FYI: Congress to allow email charges

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


I know this is off topic however a change like this could make this type of
forum impossible.





} Date: Tue, 8 Feb 2000 08:13:53 -0700
} X-Sender: liperez-at-cnmailsvr.nmsu.edu
} Mime-Version: 1.0
} To: all-at-biology.nmsu.edu
} From: liperez-at-NMSU.Edu (LSPSaldana)
} Subject: Congress to allow email charges
} Sender: owner-all-at-biology.nmsu.edu
} Precedence: bulk
} X-Keywords:
} X-UID: 43
} Status: O
}
}
} } } Congress to allow email charges
} } }
} } } Please pass this on to all you know since many of us use e-mail for
} } } business and to keep up with friends and family, I thought you'd like
} } } to know the following.
} } }
} } } Please jump on it right away and forward this to others.
} } }
} } } CNN has reported that within the next two weeks Congress is going to
} } } vote on allowing telephone companies to CHARGE A TOLL FEE for
} } } internet access.
} } } Translation: Every time we send long distance e-mail we will receive a
} } } long distance charge. This will get costly. Please visit the following web
} } } site and file a complaint. Complain to your Congressperson. We can't
} } } allow this to pass. The following address will allow you to send an
} } } e-mail on this subject DIRECTLY to your Congressperson.
} } }
} } } http://www.house.gov/writerep {http://www.house.gov/writerep}
} } }
} } } Pass this on to your friends. It is urgent! I hope all of you will pass
} } } this on to all your friends and family. We should ALL have
} } } an interest in this one.
} } }
} } } WAIT, THERE'S MORE. IN ADDITION, The last few months have revealed an
} } } alarming trend in the Government of the United States attempting to
} } } quietly push through legislation that will affect your use of the
} } Internet.
} } } Under
} } } proposed legislation the U.S. Postal Service will be attempting to bill
} } } email users out of "alternate postage fees". Bill 602P will permit the
} } } Federal Govt. to charge a 5 cent surcharge on every email delivered, by
} } } billing Internet Service Providers at source. The consumer would then be
} } } billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is
} } } working without pay to prevent this legislation from becoming law. The
} } U.S.
} } } Postal Service is claiming that lost revenue due to the proliferation of
} } } email
} } } is costing nearly $230,000,000 in revenue per year. You may have noticed
} } } their recent ad campaign "There is nothing like a letter".
} } } Since the average citizen received about 10 pieces of email per day in
} } } 1998,
} } } the cost to the typical individual would be an additional 50 cents per
} } } day, or over $180 dollars Per year, above and beyond there regular
} } Internet
} } } costs.
} } } Note that this would be money paid directly to the U.S. Postal Service
} } } for a service they do not even provide. The whole point of the Internet is
} } } democracy and non-interference. If the federal government is permitted
} } } to tamper with our liberties by adding a surcharge to email, who knows
} } Where
} } } it will end. You are already paying an exorbitant price for snail mail
} } } because of bureaucratic inefficiency. It currently takes up to 6 days for
} } a
} } } letter to be delivered from New York to Buffalo. If The U.S. Postal
} } Service
} } } is
} } } allowed to tinker with email; it will mark the end of the "free"
} } } Internet in the United States.
} } } One congressman, Tony Schnell has even suggested a "twenty to forty
} } } dollar per month surcharge on all internet service" above and beyond the
} } } government's proposed email Charges. Note that most of the major
} } } newspapers have ignored the story, the only exception being the
} } } Washingtonian
} } } which called the idea of email surcharge "a useful concept who's time has
} } } come"
} } } (March 6, 1999) Editorial.
} } }
} } } Don't sit by and watch your freedoms erode away! Send this e-mail to
} } } EVERYONE on your list, and tell all your friends and relatives to write
} } } to their congressman and say "No!" to Bill 602P.
} } }
} } }
} } } It will only take a few moments of your time, and could very well be
} } } instrumental in killing a bill we don't want.
} } }
} } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO
} } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD
} } } NOW, NOT AFTER
} } }
} } }
} } }
} } }
} } }
} } }
} }
}






From: S.A.Gusev :      gusev-at-ipm.sci-nnov.ru
Date: Thu, 10 Feb 2000 10:51:57 +0300
Subject: Solid State Electron Emitters

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Donald,
the Institute of Applied Physics
(http://www.sandy.ru/science/science/appl.new/frames.html)
constructs, produces and uses solid state high energy electron gun for
the High-power electronics and plasma physics researches.
May be they will help you.

Best regards,
Dr. S.A.Gusev


********************************************
* Institute for Physics of Microstrutures *
* Russian Academy of Science *
* ( IPM RAS ) *
* *
* Niznii Novgorod, GSP-105 *
* 603600 *
* RUSSIA *
* *
********************************************

tel: (+7)-8312-675313
fax: (+7)-8312-675553
e-mail: gusev-at-ipm.sci-nnov.ru






From: Erdem Yasar :      yasar-at-turkuaz.kku.edu.tr
Date: Thu, 10 Feb 2000 11:29:38 +0200 (EET)
Subject: about my water cooling system

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hello,
We have got a JEM3010 Transmission electron microscopy.I have got a
one problem about water cooling system.I dont
know,Which kind of water to used?I think so, We should use pure water for
water cooling?Besides My city water is not clean.What do you think about
this problem?
Thanks for your interest

**********************************
**********************************
** Research.Asst.ERDEM YASAR **
** University of Kirikkale **
** Department of Physics **
** TEM LABORATORY **
** 71450 KIRIKKALE/TURKEY **
** erdem.yasar-at-physics.org **
** yasar-at-science.ankara.edu.tr **
** yasar-at-turkuaz.kku.edu.tr **
**********************************
**********************************










From: Matt L Walker :      bmsmlw-at-bms.leeds.ac.uk
Date: Thu, 10 Feb 2000 10:08:40 -0000
Subject: TEM: Re Sarcomere cross-sections

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html



I am looking for TEM images that show cross-sectional views of skeletal
muscle
sarcomeres during the *contracted state*. In particular, I am
interested in
seeing the spacing of the thin filaments when they **overlap each
other** (i.e.,
when those from one side of the sarcomere overlap those from the other
side of
the sarcomere in the contracted state). In addition, I am interested in
seeing
the distribution of the thin filaments as they pass through the M line.
It
would be greatly appreciated if anyone could tell me if they know of any
research papers or review articles that contain such TEM images.

Thank you!

Izak Paul
Biological Sciences
Mount Royal College

The following papers are classics and would make good starting point.
Huxley, H.E. & Hanson, J. (1954). Nature, 173, 973-976.
Huxley, A.F. & Niedergerke, R.M. (1954). Nature, 173, 971.
Huxley, H.E. (1957). Biophys. & Biochem. Cytol. 3, 631-648.

Thin filaments 'pass through' the Z-line. Try John Squire and Pradeep
Luther
(3D reconstruction of fish Z-disc) and Mike Reedy (insect) or Bellinda
Bullard.

Matt Walker
School of Biomedical Sciences
Leeds University, UK







From: Malcolm Haswell :      malcolm.haswell-at-sunderland.ac.uk
Date: Thu, 10 Feb 2000 11:26:59 +0000
Subject: Film scanners - that old chestnut

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


I'm sorry to bring this topic up yet again, but I have just come across
the specification for a flat-bed scanner which looks as if it would be
suitable for scanning e.m. cut-film. It is the Epson Perfection 1200
Photo which includes a 5x4inch film adapter.

The UK web site address is:
http://www.epson.co.uk/sohoprod/imaging/scanner/perf1200/perf1200photo/spec.htm

and the specification is:
A4 Colour flatbed with 5"x4" film image scanner Single pass scanning
Optical resolution: 1200 x 2400 (30600 pixels/line)
Maximum output resolution of 9600x9600 dpi
36 bits per pixel in colour (24 bit output)
12 bits per pixel in black (8 bit output)
Optical Density 3.2D
USB (Type B)

Does anyone have any experience of this machine because it is about a
fifth of the price of the Umax flatbed film scanners? My only
reservation is that it is a SOHO product rather than professional.


Thanks in advance

Malcolm

Malcolm Haswell
Electron Microscopy
School of Sciences
Fleming Building
University of Sunderland
SUNDERLAND SR1 3SD
Tyne and Wear
UK

Tel (0191) 515 2872
e-mail: malcolm.haswell-at-sunderland.ac.uk




From: Dr. Gary Gaugler :      gary-at-gaugler.com
Date: Thu, 10 Feb 2000 05:34:15 -0600
Subject: Re: FYI: Congress to allow email charges

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


This is a recurring post about nothing more than a hoax.

Search for internet hoaxes and so forth and you will find
this one and many other interesting yet equally invalid assertions.

gary g.


At 10:58 PM 2/9/00 , you wrote:
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From: Jonathan Barnard :      Jonathan.Barnard-at-Angstrom.UU.SE
Date: Thu, 10 Feb 2000 13:38:12 +0100
Subject: coherence and spot size

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


The statement that the smallest spot size gives the best coherence can be a
misleading. The full statement should be the spot size with the smallest
angular distribution should be used. Anyone who has tried to do off-axis
holography in nanoprobe mode will testify to this.
The spatial coherence envelope, roughly the distance over which the beam is
considered 'coherent', is the Fourier transform of the angular distribution
of intensity at the source (Van Cittert Vernike theorem).

Born & Wolf (section 10.4.2)
"Hence if the linear dimensions of the source and the distance between P1
and P2 (points in the imaging plane) are small compared to the distance of
these points from the source, the degree of coherence, |mu_12| is equal to
the absolute value of the normalized Fourier transform of the intensity
function of the source"

The reason holography and high resolution work is done with te most
parallel beam possible becomes clear, the angular distribution of a
converged (or diverged beam) is wide enough to reduce the spatial coherence
envelope. In off-axis holography this is even more stringent since two
interfering points (reference wave & object wave) can be hundreds of
nanometers, microns even, apart. Elliptical illumination is used to
preserve coherence in the one important direction (minimum angular width)
and converged in the other to improve the intensity.


********************************************************
Dr Jonathan Barnard

Analytical Materials Physics
The Angstrom Laboratory, Uppsala University
P O Box 534, SE-751 21 Uppsala, Sweden
Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
http://www.angstrom.uu.se/analytical/home.html

********************************************************





From: EvexAnalyt-at-aol.com
Date: Thu, 10 Feb 2000 07:49:14 EST
Subject: Re: FYI: Congress to allow email charges

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Christine,

I read the following just the other day, from the US Postal Service

http://www.usps.gov/news/press/99/99045new.htm


FOR IMMEDIATE RELEASE
May 21, 1999
Release No. 45

E-MAIL RUMOR COMPLETELY UNTRUE

WASHINGTON – A completely false rumor concerning the U.S. Postal Service is
being circulated on Internet e-mail. As a matter of fact, the Postal Service
has learned that a similar hoax occurred recently in Canada concerning Canada
Post.

The e-mail message claims that a "Congressman Schnell" has introduced "Bill
602P" to allow the federal government to impose a 5-cent surcharge on each
e-mail message delivered over the Internet. The money would be collected by
Internet Service Providers and then turned over to the Postal Service.

No such proposed legislation exists. In fact, no "Congressman Schnell" exists.

The U.S. Postal Service has no authority to surcharge e-mail messages sent
over the Internet, nor would it support such legislation.

-30-


Evex Analytical
X-ray Analyzers and Digital Imaging Systems
857 StateRoad
Princeton, NJ 08540
609-252-9192 T
609-252-9091 F
www.evex.com
info-at-evex.com




In a message dated 2/10/00 12:26:56 AM Eastern Standard Time,
chbennet-at-nmsu.edu writes:

{ { Subj: FYI: Congress to allow email charges
Date: 2/10/00 12:26:56 AM Eastern Standard Time
From: chbennet-at-nmsu.edu (Christina bennett)
To: microscopy-at-sparc5.microscopy.com

------------------------------------------------------------------------
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
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I know this is off topic however a change like this could make this type of
forum impossible.





} Date: Tue, 8 Feb 2000 08:13:53 -0700
} X-Sender: liperez-at-cnmailsvr.nmsu.edu
} Mime-Version: 1.0
} To: all-at-biology.nmsu.edu
} From: liperez-at-NMSU.Edu (LSPSaldana)
} Subject: Congress to allow email charges
} Sender: owner-all-at-biology.nmsu.edu
} Precedence: bulk
} X-Keywords:
} X-UID: 43
} Status: O
}
}
} } } Congress to allow email charges
} } }
} } } Please pass this on to all you know since many of us use e-mail for
} } } business and to keep up with friends and family, I thought you'd like
} } } to know the following.
} } }
} } } Please jump on it right away and forward this to others.
} } }
} } } CNN has reported that within the next two weeks Congress is going to
} } } vote on allowing telephone companies to CHARGE A TOLL FEE for
} } } internet access.
} } } Translation: Every time we send long distance e-mail we will receive a
} } } long distance charge. This will get costly. Please visit the following
web
} } } site and file a complaint. Complain to your Congressperson. We can't
} } } allow this to pass. The following address will allow you to send an
} } } e-mail on this subject DIRECTLY to your Congressperson.
} } }
} } } http://www.house.gov/writerep {http://www.house.gov/writerep}
} } }
} } } Pass this on to your friends. It is urgent! I hope all of you will pass
} } } this on to all your friends and family. We should ALL have
} } } an interest in this one.
} } }
} } } WAIT, THERE'S MORE. IN ADDITION, The last few months have revealed an
} } } alarming trend in the Government of the United States attempting to
} } } quietly push through legislation that will affect your use of the
} } Internet.
} } } Under
} } } proposed legislation the U.S. Postal Service will be attempting to bill
} } } email users out of "alternate postage fees". Bill 602P will permit the
} } } Federal Govt. to charge a 5 cent surcharge on every email delivered, by
} } } billing Internet Service Providers at source. The consumer would then be
} } } billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is
} } } working without pay to prevent this legislation from becoming law. The
} } U.S.
} } } Postal Service is claiming that lost revenue due to the proliferation of
} } } email
} } } is costing nearly $230,000,000 in revenue per year. You may have noticed
} } } their recent ad campaign "There is nothing like a letter".
} } } Since the average citizen received about 10 pieces of email per day in
} } } 1998,
} } } the cost to the typical individual would be an additional 50 cents per
} } } day, or over $180 dollars Per year, above and beyond there regular
} } Internet
} } } costs.
} } } Note that this would be money paid directly to the U.S. Postal Service
} } } for a service they do not even provide. The whole point of the Internet
is
} } } democracy and non-interference. If the federal government is permitted
} } } to tamper with our liberties by adding a surcharge to email, who knows
} } Where
} } } it will end. You are already paying an exorbitant price for snail mail
} } } because of bureaucratic inefficiency. It currently takes up to 6 days for
} } a
} } } letter to be delivered from New York to Buffalo. If The U.S. Postal
} } Service
} } } is
} } } allowed to tinker with email; it will mark the end of the "free"
} } } Internet in the United States.
} } } One congressman, Tony Schnell has even suggested a "twenty to forty
} } } dollar per month surcharge on all internet service" above and beyond the
} } } government's proposed email Charges. Note that most of the major
} } } newspapers have ignored the story, the only exception being the
} } } Washingtonian
} } } which called the idea of email surcharge "a useful concept who's time has
} } } come"
} } } (March 6, 1999) Editorial.
} } }
} } } Don't sit by and watch your freedoms erode away! Send this e-mail to
} } } EVERYONE on your list, and tell all your friends and relatives to write
} } } to their congressman and say "No!" to Bill 602P.
} } }
} } }
} } } It will only take a few moments of your time, and could very well be
} } } instrumental in killing a bill we don't want.
} } }
} } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO
} } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD
} } } NOW, NOT AFTER
} } }
} } }
} } }
} } }
} } }
} } }
} }
}





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Date: Wed, 9 Feb 2000 21:58:18 -0700
To: microscopy-at-sparc5.microscopy.com
From: Christina bennett {chbennet-at-nmsu.edu}
Subject: FYI: Congress to allow email charges
Errors-to: Microscopy-request-at-sparc5.Microscopy.Com

} }




From: White, Woody N :      Woody.N.White-at-mcdermott.com
Date: Thu, 10 Feb 2000 07:14:00 -0600
Subject: Re:FYI: Congress to allow email charges -Hoax?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Is this yet another hoax eating time & bandwidth by being posted without
confirmation (like the last 3-4 times I saw this message in the pas year or
two), or is it real?

I could find no reference to it at the FCC site.... Those people who are
REALLY
in charge of telecommunication regulations....

Woody

New format, more pix:
http://www.geocities.com/capecanaveral/3722




From: White, Woody N :      Woody.N.White-at-mcdermott.com
Date: Thu, 10 Feb 2000 07:29:00 -0600
Subject: Noise vs. Be

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hello Harry,

Weelll... For most systems, if the low end noise is not inhibited (most
are),
the noise generated, apparent x-ray intensity, tends to increase with lower
ev.
This would not produce a gaussian-like peak, but a monotonic rise in
intensity
with lower ev. OTOH, pulse processor circuitry *could* be designed/setup to
produce such an effect. Seeing Be x-rays from 100% element is difficult to
impossible with most EDS systems. If compounded, the odds get worse.

Woody





From: Alwyn Eades :      jae5-at-lehigh.edu
Date: Thu, 10 Feb 2000 08:29:03 -0500
Subject: Re: Solid State Electron Emitters

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


In the early 1980's Philips believed that their research labs had
successfully developed a new electron source suitable for electron
microscopes. Indeed there was a time when they were planning to sell
microscopes with the new sources. I never heard what went wrong. There
were rumors that the lifetime was not good enough.

Anyone who wants details can find them in their library:
"An Efficient Silicon Cold Cathode for High Current Densities"
Van Gorkom and Hoeberechts
Philips Journal of Research 39 (1984) 51-60


Alwyn Eades
Department of Materials Science and Engineering
Lehigh University
5 East Packer Avenue
Bethlehem
Pennsylvannia 18015-3195
Phone 610 758 4231
Fax 610 758 4244
jae5-at-lehigh.edu





From: David_Bell-at-Millipore.com
Date: Thu, 10 Feb 2000 08:29:28 -0500
Subject: Re: FYI: Congress to allow email charges

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html




Hi all,

This, of course, is a classic internet hoax. I refer you to the following
website:


http://ciac.llnl.gov/ciac/CIACHoaxes.html#internetcharge

This hoax is designed, like many of the others, to eat up bandwidth and get
people involved in a general uproar.

Hopefully, no one panicked here! It may be worth the time to bookmark this hoax
site, or a similar one, so that when
we get messages such as this in our email, we can check their validity, before
contributing to its spread.
No insult intended towards anyone who falls victim to these emails, as I'm sure
most of us have at one time
or another.

Sincerely,

David Bell
Scientist
Electron Microscopy Lab
Millipore Corporation
80 Ashby Road
Bedford, MA 01730
(781) 533-2108






From: Scott Wight :      scott.wight-at-nist.gov
Date: Thu, 10 Feb 2000 08:54:18 -0500
Subject: Re: FYI: Congress to allow email charges

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


This is one of those internet hoaxes. It is not true, do not send it to
your friends, do not write your congressperson. Mining Co has a great web
page that debunks these hoaxes and myths
{http://urbanlegends.miningco.com/culture/urbanlegends/library/blhoax.htm?pid=27
33&cob=home} and is a good place to check before sending on any email that
urges you to send it to everyone you know. This message is a combination
of two old and popular hoaxes, see
{http://urbanlegends.miningco.com/culture/urbanlegends/library/blemtax2.htm}
and
{http://urbanlegends.about.com/culture/urbanlegends/library/weekly/aa012099.htm}
Hopefully this will die here.
Scott


} } } } Congress to allow email charges
} } } }
} } } } Please pass this on to all you know since many of us use e-mail for
} } } } business and to keep up with friends and family, I thought you'd like
} } } } to know the following.
} } } }
} } } } Please jump on it right away and forward this to others.
} } } }
} } } } CNN has reported that within the next two weeks Congress is going to
} } } } vote on allowing telephone companies to CHARGE A TOLL FEE for
} } } } internet access.
} } } } Translation: Every time we send long distance e-mail we will receive a
} } } } long distance charge. This will get costly. Please visit the following web
} } } } site and file a complaint. Complain to your Congressperson. We can't
} } } } allow this to pass. The following address will allow you to send an
} } } } e-mail on this subject DIRECTLY to your Congressperson.
} } } }
} } } } http://www.house.gov/writerep {http://www.house.gov/writerep}

..sniped...
} } } }
} } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO
} } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD
} } } } NOW, NOT AFTER


-------------------note: new mailing address----------------------
Scott Wight e-mail: scott.wight-at-nist.gov
NIST 222/A113 W voice: 301-975-3949
100 Bureau Dr STOP 8371 | fax: 301-417-1321
Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed
is my own and does not represent those of my employer.






From: William Janssen :      bjanssen-at-neuro.mssm.edu
Date: Thu, 10 Feb 2000 10:31:03 -0400
Subject: position available

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


A technical position is available in the Neurobiology of Aging program at the
Mount Sinai School of Medicine in New York. We are seeking an experienced
Electron Microscopist with a BS/BA or MS in Biology/Life Sciences. Applicants
should have excellent communication and organizational skills, an understanding
of basic laboratory procedures, and the ability to manage a large and varied
workload. The successful candidate will participate in ultrastructural studies
focusing on the effects of: estrogen and aging on hippocampal circuitry
which is implicated in learning and memory, quantitative excitatory amino
acid receptor (NMDA and AMPA) distribution within the central nervous
system of transgenic models, as well as manipulated primate and rodent
models. Qualifications include at least 2 years of experience in routine
transmission electron microscopy procedures,
ultramicrotomy, immunogold labelling, specimen preparation, digital
photography, and routine maintenance of equipment. Experience with
immunofluorescence and confocal microscopy is an asset.

We offer a salary commensurate with experience and excellent benefits. For
consideration, please mail/email your resume to:

Bill Janssen
Neurobiology of Aging
Box 867, EB9-02
Mount Sinai School of Medicine,
One Gustave L. Levy Place,
New York, NY 10029-6574

We are an equal opportunity employer fostering diversity in the workplace.
Bill Janssen
Neurobiology of Aging Laboratories
Mount Sinai School of Medicine, Box 1639
One Gustave L. Levy Place
New York, NY 10029

Phone 212-824-8789
Fax 212-849-2510





From: A.Walker :      Alan.Walker-at-sheffield.ac.uk
Date: Thu, 10 Feb 2000 14:55:24 +0000
Subject: Re: about my water cooling system

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hi,

I think the best solution would be to use a water recirculatory
system. Fill it with clean water and it should stay clean for a
long time. We use a 'Neslab'

Hope this helps

Alan Walker.

*********************************************
Alan Walker
Dept of Electronic and Electrical Engineering
University of Sheffield
Mappin Street
Sheffield S1 3JD
United Kingdom

Tel:+44-(0)114-2225365
Fax:+44-(0)114-2726391
alan.walker-at-shef.ac.uk
http://www.shef.ac.uk/uni/academic/D-H/eee/
*********************************************




From: Michael Bode :      mb-at-soft-imaging.com
Date: Thu, 10 Feb 2000 07:59:28 -0700
Subject: Re: Size determination of overlapping particles

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Rosemary,

this is of course a recurring theme in image analysis. There really is
no easy answer to this. Counting particles is easier as you can perhaps
separate the particles and arrive at the correct count, but if you don't
know what is occluded, you cannot measure it. However, if you can use
some information that is available to make some guesses or
extrapolations what the shape is, it can be done. A simple example: If
you look at a heap of coins, they may overlap. It is nevertheless
possible to measure the individual coins because I know that they are
all round. So I can take the "protruding" part of a coin and simply fit
a circle and that should give me the size pretty accurately.

Are ice crystals like snowflakes? Snowflakes, if I remember correctly,
have a six-fold symmetry. Can't you just measure one "branch" of a
snowflake (the one that you can see), and multiply that by 6? I am not
an expert on ice crystals (although I love to ski), so this may be
oversimplified. You may have to think about all the information you have
about ice crystals and can then perhaps make some guesses about the
occluded part.

Michael

Michael Bode, Ph.D.
Soft Imaging System Corp.
1675 Carr St., #105N
Lakewood, CO 80215
===================================
phone: (888) FIND SIS
(303) 234-9270
fax: (303) 234-9271
email: mailto:info-at-soft-imaging.com
web: http://www.soft-imaging.com
===================================



} ----------
} From: Rosemary Walsh[SMTP:RW9-at-PSU.EDU]
} Sent: Tuesday, February 08, 2000 3:24:35 PM
} To: microscopy-at-sparc5.Microscopy.Com
} Subject: Size determination of overlapping particles
} Auto forwarded by a Rule
}
------------------------------------------------------------------------
The Microscopy ListServer -- Sponsor: The Microscopy Society of America


Dear Listers,
I have a student interested in sizing ice crystals
which overlap. Any suggestions?
Rosemary






From: spatel-at-goodyear.com
Date: Thu, 10 Feb 2000 09:59:36 -0500
Subject: Liquid Nitrogen

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Fellow Subscribers;

We have a tedious task of liquid nitrogen transfer and fill-up on our
instruments such as SEMs, TEM and for cryo-ultramicrotomes. Is there any
compact unit on market which would condense nitrogen from air into liquid
form? I know that Liquid oxygen would be a problem. TIA;

Siddharth Patel





From: Eric Windsor :      Eric.Windsor-at-nist.gov
Date: Thu, 10 Feb 2000 10:10:27 -0500
Subject: Re: PIPS and milling damage

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Massimo,

I just read the two excellent replies from Scott and Phillip and fully agree
with them. Small angle cleavage technique (SACT) and the tripod polisher are
two very useful TEM prep techniques. In my experience, SACT is easier to learn
than tripod polishing but tripodding is applicable to more materials and yields
larger thin areas than does SACT.

As Phillip mentions, low energy milling in the final step of the process is
extremely important for reducing artifacts. I would finish using the lowest
possible energy that still yields effective milling.

Another approach is to use reactive ion beam etching (RIBE) or chemically
assisted ion beam etching (CAIBE). In both these techniques a reactive gas
(usually Iodine) is used. In RIBE, iodine is actually ionized and is used as
the milling gas. In CAIBE, iodine gas is introduced into the milling chamber
directly adjacent to the sample during argon ion milling. Both these
techniques have been shown to eliminate the ion beam damage for binary type
III-V compound semiconductors containing InP. Chew and Cullis (Ultramicroscopy
23/1987/175-198) also report improvements of ion milled surfaces for ternary
type III-V semiconductors containing InGaAs (materials you mentioned) using
RIBE.

I believe that Gatan offers an optional CAIBE attachment for the PIPS. This
may be the easiest thing to try first. Remember that iodine is very corrosive
and can damage ion milling parts if not used properly. Check with Gatan for
their recommendations.

Hope this helps,

Eric W.

Eric S. Windsor
Physical Scientist
NIST
100 Bureau Dr. Stop 8371
Gaithersburg, MD 20899-8371
(301) 975-3930
Fax: (301) 417-1321
Eric.Windsor-at-NIST.Gov


On Feb 9 -at- 09:43 Massimo Catalano wrote:

Dear Listservers,
we are massively working on analytical and structural TEM characterization
of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots.
Specimen preparation is of course a crucial point of our work.
We have just switched to PIPS (we used to make our samples using the Gatan
Duo Mill), and we are getting controversial results about the damage
introduced by the milling procedure.
We are perfectly aware of all the differences between the two instruments,
especially the absence of cooling stage and the higher beam current.
Has anyone performed a systematic and careful analysis to try to evaluate
the milling damage on similar materials systems. We are willing to start a
systematic work to assess this issue, but would like to know if anyone has
already done anything on this topic. Also, any collaboration will be more
than welcome.
Thanks.
Massimo



Dr. Massimo Catalano
CNR-IME
Campus Universitario
Via Arnesano
73100 Lecce - ITALY
tel: + 39 0832 322362
fax: + 39 0832 325299
email: massimo.catalano-at-ime.le.cnr.it
http://www.ime.le.cnr.it
http://www.ime.le.cnr.it/sime/sime.htm



Eric S. Windsor
Physical Scientist
NIST
100 Bureau Dr. Stop 8371
Gaithersburg, MD 20899-8371
(301) 975-3930
Fax: (301) 417-1321
Eric.Windsor-at-NIST.Gov




From: Malcolm Haswell :      malcolm.haswell-at-sunderland.ac.uk
Date: Thu, 10 Feb 2000 15:12:21 +0000
Subject: Re: Film scanners - that old chestnut

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Maureen and anyone else who's interested

I have found the basic description on the US/Canada website so I assume
its available there too - see address below:
http://www.epson.com/cam_scan/scanners/perfection1200/index.html

The price inclusive of transparency adapter appears to be about 200 UK
pounds over here. There seems to be three models:
1200s (standard print scanner - SCSI interface)
1200u (standard print scanner - USB interface)
1200photo (standard print scanner + 5x4inch transparency adapter - USB
interface)
It's the last of these that is of interest.

Malcolm Haswell
------------------------------------------
Maureen Petersen wrote:
}
} Malcolm:
}
} Will you divulge what this gem costs? Do you know if it is available in the
} US?
}
} Thank you, Maureen Petersen
} Dept Plant Pathology
} University of Florida
} Gainensville, FL 32611
}
} I'm sorry to bring this topic up yet again, but I have just come across
} the specification for a flat-bed scanner which looks as if it would be
} suitable for scanning e.m. cut-film. It is the Epson Perfection 1200
} Photo which includes a 5x4inch film adapter.
}
} The UK web site address is:
} http://www.epson.co.uk/sohoprod/imaging/scanner/perf1200/perf1200photo/spec.
} htm
}
} and the specification is:
} A4 Colour flatbed with 5"x4" film image scanner Single pass scanning
} Optical resolution: 1200 x 2400 (30600 pixels/line)
} Maximum output resolution of 9600x9600 dpi
} 36 bits per pixel in colour (24 bit output)
} 12 bits per pixel in black (8 bit output)
} Optical Density 3.2D
} USB (Type B)
}
} Does anyone have any experience of this machine because it is about a
} fifth of the price of the Umax flatbed film scanners? My only
} reservation is that it is a SOHO product rather than professional.
}
} Thanks in advance
}
} Malcolm
}
} Malcolm Haswell
} Electron Microscopy
} School of Sciences
} Fleming Building
} University of Sunderland
} SUNDERLAND SR1 3SD
} Tyne and Wear
} UK
}
} Tel (0191) 515 2872
} e-mail: malcolm.haswell-at-sunderland.ac.uk




From: David Knecht :      knecht-at-uconn.edu
Date: Thu, 10 Feb 2000 10:23:04 -0500
Subject: Color vs. monochrome cameras

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


I am considering the purchase of a new digital camera. Some of the new
cameras like the Spot-RT and Magnafire have the option of running as three
shot color or one shot gray-scale acquisition. If someone wants to do dual
channel fluorescence like FITC/Rhod or annexin/PI, we can do that as either
acquiring each image with a separate filter cube and combining in software,
or designing a dual filter cube and acquiring in color (or I guess
acquiring separate color images iwth the single cubes and combining??).
What is people real world experience with the relative merits of these two
approaches. It seems that most of what I read is done by separate filter
cubes combined in software, but there also seems to be a push toward fast
cooled color cameras. The advantage I can see to the color approach would
be using a color chip camera you get multiple channels simultaneously.
However, do you have to expose significantly longer to acheive this
compared to multiple gray scale acquisitions? I can't see an obvious
advantage to the three shot color approach because I suspect the light
throughput is lower and the filters sets I suspect are expensive to add on
if you already have the single fluor filters. The only real advantage I
see to three shot acquisition is the ability to easily image
stained/histology transmitted light slides and not for fluorescence. Any
thoughts appreciated. Thanks- Dave


************************************************************

Dr. David Knecht
Department of Molecular and Cell Biology
University of Connecticut
75 N. Eagleville Rd. U-125
Storrs, CT 06269
Knecht-at-uconnvm.uconn.edu
860-486-2200 860-486-4331 (fax)
home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
************************************************************






From: Kalman Rubinson :      kr4-at-is2.nyu.edu
Date: Thu, 10 Feb 2000 10:38:48 -0500 (EST)
Subject: Re: FYI: Congress to allow email charges

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


This nonsense resurfaces every few months. There has
never been any truth to it. (Of course, there's no
assurance that Congress will not act so foolishly in the
future.)

Kal

On Wed, 9 Feb 2000, Christina bennett 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 know this is off topic however a change like this could make this type of
} forum impossible.
}
}
}
}
}
} } Date: Tue, 8 Feb 2000 08:13:53 -0700
} } X-Sender: liperez-at-cnmailsvr.nmsu.edu
} } Mime-Version: 1.0
} } To: all-at-biology.nmsu.edu
} } From: liperez-at-NMSU.Edu (LSPSaldana)
} } Subject: Congress to allow email charges
} } Sender: owner-all-at-biology.nmsu.edu
} } Precedence: bulk
} } X-Keywords:
} } X-UID: 43
} } Status: O
} }
} }
} } } } Congress to allow email charges
} } } }
} } } } Please pass this on to all you know since many of us use e-mail for
} } } } business and to keep up with friends and family, I thought you'd like
} } } } to know the following.
} } } }
} } } } Please jump on it right away and forward this to others.
} } } }
} } } } CNN has reported that within the next two weeks Congress is going to
} } } } vote on allowing telephone companies to CHARGE A TOLL FEE for
} } } } internet access.
} } } } Translation: Every time we send long distance e-mail we will receive a
} } } } long distance charge. This will get costly. Please visit the following web
} } } } site and file a complaint. Complain to your Congressperson. We can't
} } } } allow this to pass. The following address will allow you to send an
} } } } e-mail on this subject DIRECTLY to your Congressperson.
} } } }
} } } } http://www.house.gov/writerep {http://www.house.gov/writerep}
} } } }
} } } } Pass this on to your friends. It is urgent! I hope all of you will pass
} } } } this on to all your friends and family. We should ALL have
} } } } an interest in this one.
} } } }
} } } } WAIT, THERE'S MORE. IN ADDITION, The last few months have revealed an
} } } } alarming trend in the Government of the United States attempting to
} } } } quietly push through legislation that will affect your use of the
} } } Internet.
} } } } Under
} } } } proposed legislation the U.S. Postal Service will be attempting to bill
} } } } email users out of "alternate postage fees". Bill 602P will permit the
} } } } Federal Govt. to charge a 5 cent surcharge on every email delivered, by
} } } } billing Internet Service Providers at source. The consumer would then be
} } } } billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is
} } } } working without pay to prevent this legislation from becoming law. The
} } } U.S.
} } } } Postal Service is claiming that lost revenue due to the proliferation of
} } } } email
} } } } is costing nearly $230,000,000 in revenue per year. You may have noticed
} } } } their recent ad campaign "There is nothing like a letter".
} } } } Since the average citizen received about 10 pieces of email per day in
} } } } 1998,
} } } } the cost to the typical individual would be an additional 50 cents per
} } } } day, or over $180 dollars Per year, above and beyond there regular
} } } Internet
} } } } costs.
} } } } Note that this would be money paid directly to the U.S. Postal Service
} } } } for a service they do not even provide. The whole point of the Internet is
} } } } democracy and non-interference. If the federal government is permitted
} } } } to tamper with our liberties by adding a surcharge to email, who knows
} } } Where
} } } } it will end. You are already paying an exorbitant price for snail mail
} } } } because of bureaucratic inefficiency. It currently takes up to 6 days for
} } } a
} } } } letter to be delivered from New York to Buffalo. If The U.S. Postal
} } } Service
} } } } is
} } } } allowed to tinker with email; it will mark the end of the "free"
} } } } Internet in the United States.
} } } } One congressman, Tony Schnell has even suggested a "twenty to forty
} } } } dollar per month surcharge on all internet service" above and beyond the
} } } } government's proposed email Charges. Note that most of the major
} } } } newspapers have ignored the story, the only exception being the
} } } } Washingtonian
} } } } which called the idea of email surcharge "a useful concept who's time has
} } } } come"
} } } } (March 6, 1999) Editorial.
} } } }
} } } } Don't sit by and watch your freedoms erode away! Send this e-mail to
} } } } EVERYONE on your list, and tell all your friends and relatives to write
} } } } to their congressman and say "No!" to Bill 602P.
} } } }
} } } }
} } } } It will only take a few moments of your time, and could very well be
} } } } instrumental in killing a bill we don't want.
} } } }
} } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO
} } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD
} } } } NOW, NOT AFTER
} } } }
} } } }
} } } }
} } } }
} } } }
} } } }
} } }
} }
}
}
}







From: David_Bell-at-Millipore.com
Date: Thu, 10 Feb 2000 10:50:30 -0500
Subject: Information on RG Lee SEM's

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html




Colleagues,

Someone at a remote location within our company has decided to purchase an RG
Lee SEM. I am looking for information and opinions from people who have one of
these microscopes. I'm interested in any service issues, reliability issues and
usability concerns that anyone may have. The model they are interested in is
the PSM 75LS. Also, if anyone has the URL for their website, that would be
appreciated, as well.

Thanks in advance,

David Bell
Scientist
Electron Microscopy Lab
Millipore Corporation
80 Ashby Road
Bedford, MA 01730
(781) 533-2108






From: Hans Dijkstra :      hans.dijkstra-at-edax.nl
Date: Thu, 10 Feb 2000 17:00:49 +0100
Subject: RE: Be X-ray peaks -I don't think so

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Scott,

A poster does not mean anything. I have seen them with the Lithium-Ka energy
listed, although quantum physics does forbid this X-ray line. But I can
assure you the Be-K line does exist.

On modern EDX systems Be can be detected, provided you have either a
windowless detector or a detector with a modern polymer type window (SUTW,
SATW, Norvar, or whatever the EDX supplier calls it. My apologies if I break
any trade-marks here...).
On most demonstration setups your EDS specialist will be pleased to show you
a Be peak. But probably from a pure Be sample only. The absorption
coefficient (MAC) of Be in basically any matrix is so huge, and the X-ray
fluorescence yield so small, that in any compound with less than 50 atomic
percent Be you will not detect a visible Be peak. For this reason most EDX
installations "in the field" are not setup to detect Be-K radiation.

WDS has a better P/B ratio, so if you have the proper multilayer crystal you
can get results. But peak shifts and peak shape alterations will make
quantification extremely difficult, not to mention the fact that for most
MACs of Be-K in any matrix we only have a ball-park figure.

Other techniques, like PEELS, are much more suitable for Li and Be analysis.
Best regards,

Hans Dijkstra

Disclaimer: This is my opinion, and not necessarily the one from EDAX Inc.
-------------------------------------------------------------
EDAX Europe www.edax.com
Ringbaan Noord 103 Tel.: +31-13-5364000
P.O. Box 4144 Fax.: +31-13-5356279
5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl
the Netherlands
-------------------------------------------------------------


} -----Original Message-----
} From: Walck. Scott D. [mailto:walck-at-ppg.com]
} Sent: Tuesday, February 08, 2000 6:57 PM
} To: 'microprobe'; 'harry.ekstrom-at-honeywell.com'
} Cc: 'Microscopy'
} Subject: RE: Be X-ray peaks -I don't think so
}
}
} ------------------------------------------------------------------------
} 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 wrong. I guess I should have looked up at the X-ray periodic table
} above my head and saw the 0.108 keV value for Be. The reason that I was
} wrong was that I did not consider the solid state aspects. I am still
right
} about not having a 2s1/2 to a 1s1/2 transition and that an X-ray carries
} away 1h-bar of angular momentum. What I didn't do was think about the
band
} structure of a solid. If you have the solid, the 2p bands of Be most be
} spilling into or be the conduction band for the electrons. That must be
the
} source for the electronic transition. I guess I should have engaged brain
} before typing.
} My most humble apologies.
} -Scott
}
} Scott D. Walck, Ph.D.
} PPG Industries, Inc.
} Glass Technology Center
} Guys Run Rd. (packages)
} P. O. Box 11472 (letters)
} Pittsburgh, PA 15238-0472
}
} Walck-at-PPG.com
}
} (412) 820-8651 (office)
} (412) 820-8161 (fax)
}
}
} "The opinions expressed are those of Scott D. Walck and not of PPG
} Industries, Inc. nor of any PPG-associated companies."
} --
}





From: Dr. Gary Gaugler :      gary-at-gaugler.com
Date: Thu, 10 Feb 2000 08:08:10 -0800
Subject: urban myths and legends - Congress to allow e-mail charges

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Here are two URLs showing that this is just another hoax;


http://www.urbanmyths.com/email_internettax602p.html



http://www.ulrc.com.au/html/report.asp?CaseFile=ULRR0027&Page=1&View=Request

gary g.





From: sherwood-at-helix.mgh.harvard.edu (Peggy Sherwood)
Date: Thu, 10 Feb 2000 11:16:00 -0500
Subject: New England Society for Microscopy-March 8th Meeting Announcement

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


To all:

The first meeting of the millenium for NESM (New England Society for
Microscopy) will be held on
March 8, 2000 at Polaroid Corporation in Wayland, MA. Registration will
begin at 5:00pm with tours
of the (new) facility following at 5:30pm. Attendees will also be asked to
fill outa questionnaire (optional) on "Professional Imaging" at this time.

A buffet supper will be served from 6-7pm followed by three scientific
presentations. The speakers
are : Gabriel Rojano, Staff Reliability Engineer-MIA-Com (Tyco Electronics
Company), Paul Bain from the Harvard Medical Library and Dr. Hjalmar
Brismer from the Karolinska Institute in Stockhold, Sweden.

Advance registration is encouraged (by March 3rd). The registration fee
for members is $5.00 and $20.00 for non-members (this includes a current
one-year membership in the society). Interested parties should contact:
Peggy Sherwood, Corresponding Secretary (MESnesm-at-aol.com).








From: Dr. Gary Gaugler :      gary-at-gaugler.com
Date: Thu, 10 Feb 2000 08:29:18 -0800
Subject: Re: Congress to allow e-mail charges

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Different subject title in case other posting got filtered out.

} Date: Thu, 10 Feb 2000 08:08:10 -0800
} To: Microscopy-at-MSA.Microscopy.Com
} From: "Dr. Gary Gaugler" {gary-at-gaugler.com}
} Subject: urban myths and legends - Congress to allow e-mail charges
}
} Here are two URLs showing that this is just another hoax;
}
}
} http://www.urbanmyths.com/email_internettax602p.html
}
}
}
} http://www.ulrc.com.au/html/report.asp?CaseFile=ULRR0027&Page=1&View=Request
}
} gary g.





From: donald j marshall :      dmrelion-at-world.std.com
Date: Thu, 10 Feb 2000 11:48:39 -0500 (EST)
Subject: Congress to allow email charges

Contents Retrieved from Microscopy Listserver Archives
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The insidious nature of this hoax/scam/... is that not only does the
original hoax get circulated, using up bandwidth, but then the replies of
the people who identified the hoax, many with the full hoax message
attached, also get circulated. And then the triple play (baseball and spring
training is near) is when people like me comment on the aforesaid. Please,
please don't reply to this note (which should have no attachments!) and make
it a home run!

Don Marshall

Donald J. Marshall
Relion Industries
P.O. Box 12
Bedford, MA 01730
Ph: 781-275-4695
FAX: 781-271-0252
email dmrelion-at-world.std.com

Cathodoluminescence, mass spectroscopy, electron beam technology


"A weed is a flower out of place."

(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)




From: Laurie Wallin :      lwallin-at-ucsd.edu
Date: Thu, 10 Feb 2000 09:00:52 -0800
Subject: RE: FYI: Congress to allow email charges

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


This is not true. It is an internet hoax. Here's the info from an urban
legends website:
---------------------------
Claim: Congress will soon be voting on whether or not your phone company
will be
allowed to charge you long-distance rates for accessing the Internet.

Status: False.

Example: [Collected on the Internet, 1999]

Please forward this to everyone you can...

There is a new bill in the US Congress that will affect ALL INTERNET
USERS. CNN stated that the Government would in two weeks time
decide whether to allow or not allow a Charge to YOUR phone bill equal
to a long distance call each time you access the Internet.

This affects us all! We cannot allow this to happen! Please visit the
following URL and fill out the necessary form to let your Congressman
know how you feel! The address is http://www.house.gov/writerep/.
Write
your representative!

Synopsis: Neither the FCC nor Congress is considering -- much less
voting on --
legislation that would impose (or allow phone companies to impose)
per-minute access fees
on Internet users. Recent decisions by the FCC in this area have dealt
only with the issue of
how phone companies reimburse each other for handling calls placed to
Internet service
providers, not with the prospect of allowing phone companies to charge
their customers
long-distance rates for Internet use.

Origins: As soon as we get hold of something we really like at a
reasonable price, we start
worrying that it will be banned, taxed, or made too expensive for us to
afford. The Internet is
no exception, as our old friend -- the capitalized, exclamation pointed,
"send this to everyone
you know" anonymous e-mail message -- is here to tell you.

Way back when in 1987, the Federal Communications Commission did
consider imposing a
surcharge for transmitting data over the public telephone network, but
they ultimately
rejected the idea (thanks in part to the more than 10,000 letters of
complaint they received).
Unable to believe our good fortune (the government wasn't going to make
us pay through
the nose for dialing up all those neat computer bulletin boards we'd
discovered), we couldn't
leave well enough alone, and in 1991 a flood of urgent messages warning
us that the FCC
was again considering a proposal they'd rejected three years earlier hit
e-mail systems all
over the country (and the nascently popular Internet). Like the
ubiquitous Craig Shergold
message, the "modem tax" warning would long outlive the validity of the
information it
conveyed.

Fast forward to 1998. On-line services, the Internet, the World Wide
Web, e-mail, and chat
rooms are more popular than ever, a daily part of many people's lives.
Somebody -- the
government, the phone companies, Bill Gates, the Grinch -- must be on
his way to spoil the
party. Sure enough, we're now being told the phone companies and the
government are in
cahoots to ruin our good time.

First of all, a little background. Most of us still have to dial up over
a modem and connect to
an Internet Service Provider (ISP) to access the Internet. If your ISP
is in your local dialing
area, you probably don't pay anything at all for the call, no matter how
long you stay
connected. This means you get to tie up a phone line with your modem for
hours and hours
on end, every day, at no charge beyond the price of basic phone service.
And the party at the
other end of the line -- your ISP -- isn't paying anything extra,
either. It's easy to see that
somebody has a chance to reap some windfall profits here. If the phone
companies were
allowed to charge you a per-minute fee for accessing the Internet (or
the government were
allowed to tax your use of the Internet), their coffers would soon
overflow with cash.

Scary thought, isn't it? All the phone companies need, we hear, is to
get the FCC to
reclassify and/or regulate ISPs, and then the phone companies can charge
gobs of extra
money for handling Internet traffic. And Congress is just about to vote
on that very issue,
we're told.

In fact, there is no such proposal before Congress, and there never has
been.

The only real issue before the FCC concerning Internet usage (and the
genesis of this latest
round of scaremongering) is the subject of "reciprocal compensation." In
short, reciprocal
compensation means that when you place a local call to someone who is
serviced by a
different phone company, your phone company has to compensate his phone
company for
completing the call. (On the other hand, when you place a long-distance
call, the long
distance carrier who handles the traffic has to pay access charges to
your phone company
for originating the call.) But if the "person" you're calling is an ISP,
should your phone
company have to compensate the ISP's phone company?

The subject of recpirocal compensation has been a hot issue of late
because new local phone
companies have been springing up all over the place. The bigger phone
companies, figuring
that they would have many more customers than their newer (and smaller)
competitors,
negotiated reciprocal compensation agreements with the new phone
companies. Every time
one of these little phone companies' customers placed a call to a
destination outside his local
service that ended up on the bigger phone company's network, the big
phone company
would get to collect money from the little phone company. Not a bad
scheme, the big phone
companies thought.

Ah, but some of the little guys had a neat trick up their sleeves. They
started offering their
services to Internet service providers -- ISPs with banks and banks of
modems that received
thousands and thousands of calls every day, but never made any outgoing
calls. All the
reciprocal compensation started flowing one direction, from the big
phone companies to the
little phone companies, which wasn't what the big guys had in mind at
all. "Foul," they cried.
"Internet traffic flows all over the world," they noted. "Internet
traffic is therefore interstate in
nature and should be classified as long-distance, so the little guys
should be paying us for
originating the calls," they insisted. "We're not paying," they sputtered.

Enter the FCC to resolve the dispute, which they did (for now) on 25
February 1999 by
ruling that phone companies are bound by whatever reciprocal
compensation agreements
they've negotiated with each other, whether they think they're fair or
not. That was the only
issue before the FCC. But most of us are already struggling with a glut
of information, and
we don't have the time to familiarize ourselves with details like
interconnection agreements
and reciprocal compensation, so when we hear reports with buzzwords like
"Internet,"
"FCC," "access fees," and "long distance," we assume the worst, even
though the real story
is something quite different. And even if we make the effort to
understand the whole story,
we find all too often that we're reading information that has been
misreported by others --
often the mainstream media -- who didn't make enough of an effort to
understand the whole
story themselves. If we can't even depend upon the people whose business
it is to supply us
with accurate information to get the facts straight, what more can we
do? (See, for example,
the misleading headline on the CNN article referenced in the "Additional
information"
section below.)

A few important points related to the recent FCC decision:

Didn't the FCC rule that Internet connections are long-distance
calls?

Sort of. The FCC declared that "Internet traffic is
jurisdictionally mixed and appears to
be largely interstate in nature," which is the technical
definition of "long distance." But
that doesn't mean -- as is often misreported -- that Internet
users will be paying long
distance rates for dial-up connections, since the Internet has
been, and still is, exempt
from interstate access charges. The FCC did nothing to abolish
that exemption.

Won't the phone companies just pass the cost of carrying Internet
traffic to
customers by raising their phone rates?

There is no guarantee that phone rates won't go up in the future,
of course. However,
since most states require phone companies to charge a flat rate
for unlimited local
usage, you won't have to pay per-minute charges for accessing the
Internet (as long as
your ISP has a dial-up number within your local service area).

What if the FCC changes their mind?

The possibility exists that the FCC might someday decide that
additional fees can be
imposed for Internet access. But as Bill Kennard, the chairman of
the FCC, has stated
on more than one occasion: "I want to say this as clearly as I can
. . . as long as I'm
chairman of the Federal Communications Commission this agency will
not regulate
the Internet. It's not going to happen. The FCC has no intention
of making computer
users pay long distance fees for dial-up access to the Internet,
as people now pay when
they make long-distance telephone calls. These rumors get on the
Internet that the big
bad FCC is going to impose all this regulation on the Internet.
Now I know this
painfully because every so often when one of these rumors flares
up I get, literally,
about 600 e-mail messages a day by people who are telling me to
keep my hands off
the Internet."
[Note: this is not a direct quote from Kennard; it is pieced
together from multiple
statements of his.]

Additional information:

No Consumer Per-Minute Charges to Access ISPs (FCC)

Users, Advertisers Await FCC Decision on Internet Charges
(CNN)

Update: In April 1999, a Canadian version of this message was
unleashed on the Internet.
Unlike its American counterpart, this version is not mere misinformation
based on a flawed
understanding of actual events or legislation -- it is an outright hoax:

Please read the following carefully if you intend to stay
online and continue using email:

The last few months have revealed an alarming trend in the
Government of Canada attempting to quietly push through
legislation that will affect your use of the Internet. Under
proposed legislation Canada Post will be attempting to bilk
email users out of "alternate postage fees".

Bill 602P will permit the Federal Govt to charge a 5 cent
surcharge on every email delivered, by billing Internet
Service Providers at source. The consumer would then be
billed in turn by the ISP. Toronto lawyer Richard Stepp QC is
working without pay to prevent this legislation from
becoming law.

The Canada Post Corporation is claiming that lost revenue
due to the proliferation of email is costing nearly
$23,000,000 in revenue per year. You may have noticed
Canada Post's recent ad campaign "There is nothing like a
letter". Since the average citizen received about 10 pieces
of email per day in 1998, the cost to the typical individual
would be an additional 50 cents per day, or over $180
dollars per year, above and beyond their regular Internet
costs. Note that this would be money paid directly to
Canada Post for a service they do not even provide. The
whole point of the Internet is democracy and
non-interference. If the Canadian Government is permitted
to tamper with our liberties by adding a surcharge to email,
who knows where it will end. You are already paying an
exhorbitant price for snail mail because of beaurocratic
inefficiency. It currently takes up to 6 days for a letter to be
delivered from Mississauga to Scarborough. If Canada Post
Corporation is allowed to tinker with email, it will mark the
end of the "free" Internet in Canada. One back-bencher,
Liberal Tony Schnell (NB) has even suggested a "twenty to
forty dollar per month surcharge on all Internet service"
above and beyond the government's proposed email
charges. Note that most of the major newspapers have
ignored the story, the only exception being the Toronto Star
that called the idea of email surcharge "a useful concept
who's time has come" (March 6th 1999 Editorial) Don't sit
by and watch your freedoms erode away! Send this email to
all Canadians on your list and tell your friends and relatives
to write to their MP and say "No!" to Bill 602P.

Kate Turner
Assistant to Richard Stepp QC
Berger, Stepp and Gorman
Barristers at Law
216 Bay Street
Toronto, ON
MlL 3C6

Yes, Americans are not alone in their belief that when the need for the
primary service
provided by a governmental agency diminishes or disappears, that agency
will come up with
draconian schemes to perpetuate its existence at taxpayer expense rather
than modernizing
or closing up shop. In this case, however, the message is simply too
riddled with errors to be
anything but a hoax:

There is no "Bill 602P" currently before the Canadian parliament.
The designations of
Canadian parliamentary bills take the form of the letter 'C' or
'S' followed by a
number (depending upon whether they originated in the House of
Commons or the
Senate). Besides having the wrong prefix, this purported bill is
assigned a number far
too high to be one currently being considered in parliament, as
you can see on the list
of Canadian government bills.

Despite the claim in the message, nothing about this alleged bill
is to be found. Also,
there was no editorial about this on the 6 March 1999 OpEd page of
Toronto
Star. (Maybe "major papers have ignored the story" because it's a
work of
fiction?)

There is no Richard Stepp QC; law firm by the name of Berger,
Stepp and Gorman; or
216 Bay Street in Toronto.

A list of Canadian Members of Parliament contains no MP by the
name of Tony
Schnell.

Of course, it didn't take long before the same thing started circulating
in an Americanized
version:

Dear Internet Subscriber:

Please read the following carefully if you intend to stay
online and continue using email: The last few months have
revealed an alarming trend in the Government of the United
States attempting to quietly push through legislation that
will affect your use of the Internet. Under proposed
legislation the U.S. Postal Service will be attempting to bilk
email users out of "alternate postage fees".

Bill 602P will permit the Federal Govt to charge a 5 cent
surcharge on every email delivered, by billing Internet
Service Providers at source. The consumer would then be
billed in turn by the ISP. Washington D.C. lawyer Richard
Stepp is working without pay to prevent this legislation
from becoming law.

The U.S. Postal Service is claiming that lost revenue due to
the proliferation of email is costing nearly $230,000,000 in
revenue per year. You may have noticed their recent ad
campaign "There is nothing like a letter". Since the average
citizen received about 10 pieces of email per day in 1998,
the cost to the typical individual would be an additional
50 cents per day, or over $180 dollars per year, above and
beyond their regular Internet costs. Note that this would be
money paid directly to the U.S. Postal Service for a service
they do not even provide. The whole point of the Internet is
democracy and non-interference. If the federal government
is permitted to tamper with our liberties by adding a
surcharge to email, who knows where it will end. You are
already paying an exorbitant price for snail mail because of
bureacratic efficiency. It currently takes up to 6 days for a
letter to be delivered from New York to Buffalo. If the U.S.
Postal Service is allowed to tinker with email, it will mark
the end of the "free" Internet in the United States. One
congressman, Tony Schnell (r) has even suggested a
"twenty to forty dollar per month surcharge on all Internet
service" above and beyond the government's proposed
email charges. Note that most of the major newspapers
have ignored the story, the only exception being the
Washingtonian which called the idea of email surcharge "a
useful concept who's time has come" (March 6th 1999
Editorial) Don't sit by and watch your freedoms erode away!

Send this email to all Americans on your list and tell your
friends and relatives to write to their congressman and say
"No!" to Bill 602P.

Kate Turner
Assistant to Richard Stepp
Berger, Stepp and Gorman
Attorneys at Law
216 Concorde Street,
Vienna, Va.




From: Laurie Wallin :      lwallin-at-ucsd.edu
Date: Thu, 10 Feb 2000 09:03:00 -0800
Subject: paraffin sectioning troubleshooting guide

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Does anyone know where I can find a troubleshooting manual/guide for
paraffin sectioning? I am having trouble with the tissue "smearing" and
with it looking chopped-up. Is this a vibration issue? I welcome any
comments from those of you with experience in this area!

Thanks in advance.

Sincerely.




From: Laurie Wallin :      lwallin-at-ucsd.edu
Date: Thu, 10 Feb 2000 09:04:17 -0800
Subject: Fwd: paraffin sectioning troubleshooting guide

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Sorry, I forgot to sign my note last time...


} Date: Thu, 10 Feb 2000 09:03:00 -0800
} To: microscopy-at-sparc5.microscopy.com
} From: Laurie Wallin {lwallin-at-ucsd.edu}
} Subject: paraffin sectioning troubleshooting guide
} Cc:
} Bcc:
} X-Attachments:
}
} Does anyone know where I can find a troubleshooting manual/guide for
} paraffin sectioning? I am having trouble with the tissue "smearing" and
} with it looking chopped-up. Is this a vibration issue? I welcome any
} comments from those of you with experience in this area!
}
} Thanks in advance.
}
} Sincerely.

Laurie Wallin
UCSD Department of Anesthesiology
9500 Gilman Drive, 0629, La Jolla, CA 92093
(858) 822-3271




From: David Henriks :      Henriks-at-CompuServe.COM
Date: Thu, 10 Feb 2000 12:31:07 -0500
Subject: PIPS and milling damage

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Dr. Catalano:

There has been some very significant work done by Dr. Arpad Barna et al in
Hungary. Some of the papers he has presented include:

Ion Energy Effect on Surface Amorphisation of Semiconductor Crystals/A.
Barna, et al

Amorphisation and Surface Morphology Development at Low Energy Ion
Milling/A. Barna, B. Pecz.

Analysis of the Development of Large Surface Topography During Ion
Etching/A. Barna; P. Barna; et al

Model Considerations of Ion Beam Thinning for Preparing TEM
Samples/A.Barna; et al

Possibility of Surface Polishing by Ion Beam Thinning/ A. Barna

Ion Beam Thinning on the Basis of Topographic Kinetics/A. Barna

Low Angle and Low Energy Ion Beam Etching for TEM Sample Preparation/A.
Barna

TEM Sample Preparation by Ion Milling / Amorphization/A. Barna, B. Pecz, M.
Menyhard

I also have the following paper that may be of interest:

Preparation of InGaAs/GaAs Multilayered Materials for TEM by One Side
Non-Rotation Ion Beam Thinning/J.Y. Yao; G.L. Dunlop

I do not have the complete references available in front of me, but we do
have copies of all of these papers in our technical library. I would be
pleased to mail copies to you if that would be of interest. We also have a
list of over 250 papers dealing with various aspects of sample preparation
(much of it TEM sample preparation) which may be of interest. If you would
like to review that list, I can send it over to you in MS Excel format and
you can select any other papers that would be of interest.

I hope this helps.

DISCLAIMER: South Bay Technology, Inc. markets the IV3 Ion Mill in both
high and low energy (down to 200ev) versions which is based on the work of
Dr. Barna. We also produce the XLA 2000 computer controlled Low Angle Ion
Mill so we have a vested interest in promoting their use.

Best regards-

David
Writing at 10:05:16 AM on 02/09/2000

***************************************************************************
************************

David Henriks
Vice President TEL: 800-728-2233
(toll free in the USA)
South Bay Technology, Inc. +1-949-492-2600
1120 Via Callejon FAX: +1-949-492-1499
San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com

***************************************************************************
************************

} } } } } Please visit us at http://www.southbaytech.com { { { { {

Manufacturers of precision sample preparation equipment and supplies for
metallography, crystallography and electron microscopy.

Message text written by Massimo Catalano
}
------------------------------------------------------------------------
The Microscopy ListServer -- Sponsor: The Microscopy Society of America


Dear Listservers,

we are massively working on analytical and structural TEM characterization
of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots.

Specimen preparation is of course a crucial point of our work.

We have just switched to PIPS (we used to make our samples using the Gatan
Duo Mill), and we are getting controversial results about the damage
introduced by the milling procedure.

We are perfectly aware of all the differences between the two instruments,
especially the absence of cooling stage and the higher beam current.

Has anyone performed a systematic and careful analysis to try to evaluate
the milling damage on similar materials systems. We are willing to start a
systematic work to assess this issue, but would like to know if anyone has
already done anything on this topic. Also, any collaboration will be more
than welcome.

Thanks.

Massimo




Dr. Massimo Catalano
CNR-IME
Campus Universitario
Via Arnesano
73100 Lecce - ITALY
tel: + 39 0832 322362
fax: + 39 0832 325299
email: massimo.catalano-at-ime.le.cnr.it
http://www.ime.le.cnr.it
http://www.ime.le.cnr.it/sime/sime.htm


{




From: Roger Moretz :      rcmoretz-at-excite.com
Date: Thu, 10 Feb 2000 09:42:05 -0800 (PST)
Subject: Re: FYI: Congress to allow email charges

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Well, this urban legend has finally made it to the microscopy listserver!
And that is just what it is--a legend, a myth, a fable. As a stamp
collector and one who keeps abreast of all of this kind of stuff, this urban
legend has been debunked in both Linn's Stamp News and Stamp Collector.

All of us have important issues to consider. Please, put this one to rest.

Roger Moretz


On Wed, 9 Feb 2000 21:58:18 -0700, Christina bennett 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 know this is off topic however a change like this could make this type
of
} forum impossible.
}
}
}
}
}
} } Date: Tue, 8 Feb 2000 08:13:53 -0700
} } X-Sender: liperez-at-cnmailsvr.nmsu.edu
} } Mime-Version: 1.0
} } To: all-at-biology.nmsu.edu
} } From: liperez-at-NMSU.Edu (LSPSaldana)
} } Subject: Congress to allow email charges
} } Sender: owner-all-at-biology.nmsu.edu
} } Precedence: bulk
} } X-Keywords:
} } X-UID: 43
} } Status: O
} }
} }
} } } } Congress to allow email charges
} } } }
} } } } Please pass this on to all you know since many of us use e-mail for
} } } } business and to keep up with friends and family, I thought you'd like
} } } } to know the following.
} } } }
} } } } Please jump on it right away and forward this to others.
} } } }
} } } } CNN has reported that within the next two weeks Congress is going to
} } } } vote on allowing telephone companies to CHARGE A TOLL FEE for
} } } } internet access.
} } } } Translation: Every time we send long distance e-mail we will receive
a
} } } } long distance charge. This will get costly. Please visit the
following web
} } } } site and file a complaint. Complain to your Congressperson. We can't
} } } } allow this to pass. The following address will allow you to send an
} } } } e-mail on this subject DIRECTLY to your Congressperson.
} } } }
} } } } http://www.house.gov/writerep {http://www.house.gov/writerep}
} } } }
} } } } Pass this on to your friends. It is urgent! I hope all of you will
pass
} } } } this on to all your friends and family. We should ALL have
} } } } an interest in this one.
} } } }
} } } } WAIT, THERE'S MORE. IN ADDITION, The last few months have revealed an
} } } } alarming trend in the Government of the United States attempting to
} } } } quietly push through legislation that will affect your use of the
} } } Internet.
} } } } Under
} } } } proposed legislation the U.S. Postal Service will be attempting to
bill
} } } } email users out of "alternate postage fees". Bill 602P will permit
the
} } } } Federal Govt. to charge a 5 cent surcharge on every email delivered,
by
} } } } billing Internet Service Providers at source. The consumer would then
be
} } } } billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is
} } } } working without pay to prevent this legislation from becoming law.
The
} } } U.S.
} } } } Postal Service is claiming that lost revenue due to the proliferation
of
} } } } email
} } } } is costing nearly $230,000,000 in revenue per year. You may have
noticed
} } } } their recent ad campaign "There is nothing like a letter".
} } } } Since the average citizen received about 10 pieces of email per day
in
} } } } 1998,
} } } } the cost to the typical individual would be an additional 50 cents
per
} } } } day, or over $180 dollars Per year, above and beyond there regular
} } } Internet
} } } } costs.
} } } } Note that this would be money paid directly to the U.S. Postal
Service
} } } } for a service they do not even provide. The whole point of the
Internet is
} } } } democracy and non-interference. If the federal government is
permitted
} } } } to tamper with our liberties by adding a surcharge to email, who
knows
} } } Where
} } } } it will end. You are already paying an exorbitant price for snail
mail
} } } } because of bureaucratic inefficiency. It currently takes up to 6 days
for
} } } a
} } } } letter to be delivered from New York to Buffalo. If The U.S. Postal
} } } Service
} } } } is
} } } } allowed to tinker with email; it will mark the end of the "free"
} } } } Internet in the United States.
} } } } One congressman, Tony Schnell has even suggested a "twenty to forty
} } } } dollar per month surcharge on all internet service" above and beyond
the
} } } } government's proposed email Charges. Note that most of the major
} } } } newspapers have ignored the story, the only exception being the
} } } } Washingtonian
} } } } which called the idea of email surcharge "a useful concept who's time
has
} } } } come"
} } } } (March 6, 1999) Editorial.
} } } }
} } } } Don't sit by and watch your freedoms erode away! Send this e-mail to
} } } } EVERYONE on your list, and tell all your friends and relatives to
write
} } } } to their congressman and say "No!" to Bill 602P.
} } } }
} } } }
} } } } It will only take a few moments of your time, and could very well be
} } } } instrumental in killing a bill we don't want.
} } } }
} } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE
TWO
} } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD
} } } } NOW, NOT AFTER
} } } }
} } } }
} } } }
} } } }
} } } }
} } } }
} } }
} }
}
}
}


Roger C Moretz, Ph.D.





_______________________________________________________
Get 100% FREE Internet Access powered by Excite
Visit http://freeworld.excite.com





From: hank adams :      hpadams-at-bcm.tmc.edu
Date: Thu, 10 Feb 2000 11:45:28 -0600
Subject: GFP and Osmium

Contents Retrieved from Microscopy Listserver Archives
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Does anyone know if GFP is quenched after OsO4 postfixation?
Hank Adams
Integrated Microscopy Core
Molecular and Cell Biology
Baylor College of Medicine
Houston, TX 77030





From: Carlos Kazuo Inoki :      kazuo-at-csc.albany.edu
Date: Thu, 10 Feb 2000 13:05:16 -0500 (EST)
Subject: TEM: sample preparation for InSb

Contents Retrieved from Microscopy Listserver Archives
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Hi,

I'm working with a InSb sample prepared with tripod polishing.
Since the layer is quite thick, more than 3 micron plus a 2 micron buffer
layer, I am experimenting some difficult to prepare a good sample for
cross section observation. The final polishing is done in a Gatan ion
milling set using a low angle. The problem with my samples is how to get a
uniform thickness from the top of the sample to the substrate. Usually the
top of the sample is milled much faster than near to the substrate. I
can't mechanically polish the sample too much, since the sample is
somewhat fragile (brittle). Even in old papers people report this kind of
problem when preparing InSb samples for TEM using ion milling. Does
someone else suffered from this kind of problem: the top part of a
sample being milled away? Oh yes, when I polish the sample I try to make
wedge perpendicular to the sample surface, so the thickness of the sample
is the same from the InSb layer to the substrate after the mechanical
polishing. There is some correlation between the ion milling angle/energy
with this effect? The Gatan epoxy I use to glue a Si piece on top of the
sample should have some influence (charging)? Thanks.

Kazuo

o-------------------------------------------------------o
| Carlos Kazuo Inoki |
| Department of Physics - University at Albany |
| 1400 Washington Ave.- Albany - NY - 12222 |
o-------------------------------------------------------o





From: Walck. Scott D. :      walck-at-ppg.com
Date: Thu, 10 Feb 2000 13:51:03 -0500
Subject: RE: Be X-ray peaks, solid state physics, how about 2nd row elemen

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I think that I know understand the reason why you get Be X-ray and why
there are energy shifts when it is in different compounds. My next question
then is do the elements between B and F have measurable energy shifts
depending on what material they are in? The key word here is measurable.
The Ka1 and Ka2 peaks are due to transitions form the 2p1/2 and 2p3/2 (LII
and LIII) to the 1s1/2 state. But these would be valence/conductance bands
for the pure elements. (Assuming you collect your X-rays from solid N2, O2,
or F2) The band structure would be different for different materials. For
example, do you see any shifts between B2O3? and BN or C(diamond) and
C(graphite) or TiC? Any microprobers following this thread?

-Scott

Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)



} -----Original Message-----
} From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl]
} Sent: Thursday, February 10, 2000 11:01 AM
} To: Walck. Scott D.; 'microprobe'
} Cc: 'Microscopy'
} Subject: RE: Be X-ray peaks -I don't think so
}
}
} --------------------------------------------------------------
} ----------
} 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 Scott,
}
} A poster does not mean anything. I have seen them with the
} Lithium-Ka energy
} listed, although quantum physics does forbid this X-ray line.
} But I can
} assure you the Be-K line does exist.
}
} On modern EDX systems Be can be detected, provided you have either a
} windowless detector or a detector with a modern polymer type
} window (SUTW,
} SATW, Norvar, or whatever the EDX supplier calls it. My
} apologies if I break
} any trade-marks here...).
} On most demonstration setups your EDS specialist will be
} pleased to show you
} a Be peak. But probably from a pure Be sample only. The absorption
} coefficient (MAC) of Be in basically any matrix is so huge,
} and the X-ray
} fluorescence yield so small, that in any compound with less
} than 50 atomic
} percent Be you will not detect a visible Be peak. For this
} reason most EDX
} installations "in the field" are not setup to detect Be-K radiation.
}
} WDS has a better P/B ratio, so if you have the proper
} multilayer crystal you
} can get results. But peak shifts and peak shape alterations will make
} quantification extremely difficult, not to mention the fact
} that for most
} MACs of Be-K in any matrix we only have a ball-park figure.
}
} Other techniques, like PEELS, are much more suitable for Li
} and Be analysis.
} Best regards,
}
} Hans Dijkstra
}
} Disclaimer: This is my opinion, and not necessarily the one
} from EDAX Inc.
} -------------------------------------------------------------
} EDAX Europe www.edax.com
} Ringbaan Noord 103 Tel.: +31-13-5364000
} P.O. Box 4144 Fax.: +31-13-5356279
} 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl
} the Netherlands
} -------------------------------------------------------------
}
}
} } -----Original Message-----
} } From: Walck. Scott D. [mailto:walck-at-ppg.com]
} } Sent: Tuesday, February 08, 2000 6:57 PM
} } To: 'microprobe'; 'harry.ekstrom-at-honeywell.com'
} } Cc: 'Microscopy'
} } Subject: RE: Be X-ray peaks -I don't think so
} }
} }
} }
} --------------------------------------------------------------
} ----------
} } 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 wrong. I guess I should have looked up at the X-ray
} periodic table
} } above my head and saw the 0.108 keV value for Be. The
} reason that I was
} } wrong was that I did not consider the solid state aspects.
} I am still
} right
} } about not having a 2s1/2 to a 1s1/2 transition and that an
} X-ray carries
} } away 1h-bar of angular momentum. What I didn't do was
} think about the
} band
} } structure of a solid. If you have the solid, the 2p bands
} of Be most be
} } spilling into or be the conduction band for the electrons.
} That must be
} the
} } source for the electronic transition. I guess I should
} have engaged brain
} } before typing.
} } My most humble apologies.
} } -Scott
} }
} } Scott D. Walck, Ph.D.
} } PPG Industries, Inc.
} } Glass Technology Center
} } Guys Run Rd. (packages)
} } P. O. Box 11472 (letters)
} } Pittsburgh, PA 15238-0472
} }
} } Walck-at-PPG.com
} }
} } (412) 820-8651 (office)
} } (412) 820-8161 (fax)
} }
} }
} } "The opinions expressed are those of Scott D. Walck and not of PPG
} } Industries, Inc. nor of any PPG-associated companies."
} } --
} }
}
}




From: Walck. Scott D. :      walck-at-ppg.com
Date: Thu, 10 Feb 2000 13:57:08 -0500
Subject: RE: sample preparation for InSb

Contents Retrieved from Microscopy Listserver Archives
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I did InSb and AlInSb by the small angle cleavage technique when I was at
Wright Patterson AFB. I assume since you are using the Tripod Technique
that you require site-specific results. If not, try it. It works really
well on this material.

I am getting to sound like a broken record on this topic, aren't I? See my
post yesterday.

-Scott



Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)


"The opinions expressed are those of Scott D. Walck and not of PPG
Industries, Inc. nor of any PPG-associated companies."
--



Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)



} -----Original Message-----
} From: Carlos Kazuo Inoki [mailto:kazuo-at-csc.albany.edu]
} Sent: Thursday, February 10, 2000 1:05 PM
} To: Microscopy-at-sparc5.microscopy.com
} Subject: TEM: sample preparation for InSb
}
}
} --------------------------------------------------------------
} ----------
} 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'm working with a InSb sample prepared with tripod polishing.
} Since the layer is quite thick, more than 3 micron plus a 2
} micron buffer
} layer, I am experimenting some difficult to prepare a good sample for
} cross section observation. The final polishing is done in a Gatan ion
} milling set using a low angle. The problem with my samples is
} how to get a
} uniform thickness from the top of the sample to the
} substrate. Usually the
} top of the sample is milled much faster than near to the substrate. I
} can't mechanically polish the sample too much, since the sample is
} somewhat fragile (brittle). Even in old papers people report
} this kind of
} problem when preparing InSb samples for TEM using ion milling. Does
} someone else suffered from this kind of problem: the top part of a
} sample being milled away? Oh yes, when I polish the sample I
} try to make
} wedge perpendicular to the sample surface, so the thickness
} of the sample
} is the same from the InSb layer to the substrate after the mechanical
} polishing. There is some correlation between the ion milling
} angle/energy
} with this effect? The Gatan epoxy I use to glue a Si piece on
} top of the
} sample should have some influence (charging)? Thanks.
}
} Kazuo
}
} o-------------------------------------------------------o
} | Carlos Kazuo Inoki |
} | Department of Physics - University at Albany |
} | 1400 Washington Ave.- Albany - NY - 12222 |
} o-------------------------------------------------------o
}
}




From: David_Bell-at-Millipore.com
Date: Thu, 10 Feb 2000 14:06:06 -0500
Subject: My apologies

Contents Retrieved from Microscopy Listserver Archives
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Hello again,

My apologies to the list and especially those at RJ Lee! Hope nobody was
offended.

David Bell
Scientist
Electron Microscopy Lab
Millipore Corporation
80 Ashby Road
Bedford, MA 01730
(781) 533-2108






From: Damian :      dneuberger-at-mindspring.com
Date: Thu, 10 Feb 2000 13:16:34 -0600
Subject: Re:FYI: Congress to allow email charges -Hoax?

Contents Retrieved from Microscopy Listserver Archives
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Woody,

I hate to add to the bandwidth clutter but these are very old hoaxes.

Damian


} Is this yet another hoax eating time & bandwidth by being posted without
} confirmation (like the last 3-4 times I saw this message in the pas year or
} two), or is it real?





From: Alexander Mironov Jr. :      amironov-at-cmns.mnegri.it
Date: Thu, 10 Feb 2000 21:54:09 +0100 (MET)
Subject: Re: GFP and Osmium

Contents Retrieved from Microscopy Listserver Archives
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Dear Hank,

Yes, after OsO4 GFP is quenched, moreover it is quenched significantly
(three times or more) even with 0.05% of glutaraldehyde (10 min.


Dr. Alexander A. Mironov Jr., MD, PhD
Unit of Morphology
Dept. of Cell Biology and Oncology
Consorzio Mario Negri Sud
Via Nazionale, S.Maria Imbaro (Ch)
66030 Italy

Tel. 0039-0872-570-332
Fax 0039-0872-570-412
E-mail: amironov-at-cmns.mnegri.it


On Thu, 10 Feb 2000, hank adams wrote:

} ------------------------------------------------------------------------
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} -----------------------------------------------------------------------.
}
}
} Does anyone know if GFP is quenched after OsO4 postfixation?
} Hank Adams
} Integrated Microscopy Core
} Molecular and Cell Biology
} Baylor College of Medicine
} Houston, TX 77030
}
}
}





From: Fagerland,Jane :      jane.a.fagerland-at-abbott.com
Date: Thu, 10 Feb 2000 15:13:19 -0600
Subject: Summer Internships at Abbott - more information

Contents Retrieved from Microscopy Listserver Archives
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Thanks to all of you who have shown an interest in our internships. I'd like
to provide a few more details and answer some questions I've received so far
about the summer internships at Abbott.

First: Are the internships open to international students? Technically,
yes. However, interns are employees of Abbott Laboratories during their time
here. Thus, the logistics of processing the necessary paperwork for non-US
citizens for a 12-week employment period would not be practical. So, in
truth, the answer has to be that, except in highly unusual circumstances, we
will limit applications to students in the United States.

Second: Are the internships intended for college or high school students?
The internships are for college students working towards a Bachelor's degree
or higher. Students in the Associate degree programs at Madison Area
Technical College and San Joaquin Delta College are eligible, but priority
will be given to students who already have a Bachelor's degree.

Third: How to apply? Go to the Abbott Laboratories website at abbott.com
and follow the links: Careers, Entry Level Positions, Summer Internship
Program. There you will find an electronic resume form that can be submitted
from your computer. THE DEADLINE FOR SUBMISSION IS MARCH 1.

Along with submitting the electronic resume, please send me either an
e-mailed or faxed copy of your resume, or a hardcopy via snail mail. The
electronic resume ends up in Corporate Staffing, where your skills will be
matched with the internship openings at Abbott. If I have a copy of your
resume, I can make sure that you are considered for positions in the
microscopy department.

If there are any other questions, please feel free to contact me.

Thanks,

Jane A. Fagerland, Ph.D.
Dept. Microscopy and Microanalysis
D-45M/AP31 200 Abbott Park Rd.
Abbott Park IL 60064-6202
voice: (847) 935-0104
fax: (847) 938-5027
e-mail: jane.a.fagerland-at-abbott.com




From: Robert Underwood :      underwoo-at-u.washington.edu
Date: Thu, 10 Feb 2000 14:04:19 -0800 (PST)
Subject: Re: autofluorescence quenching

Contents Retrieved from Microscopy Listserver Archives
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Hello from Seattle,

There was a paper by Cowen, Haven and Burnstock 1985, using Pontamine Sky
Blue couterstain to reduce autofluorescence.

Also a paper by Kittleburger, Davis, and Stephans in Acta Histochem 89,
using Eriochrome Black T.

Bob
Morphology Core
U of Washington

On Wed, 9 Feb 2000, Corazon Bucana wrote:

} ------------------------------------------------------------------------
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} -----------------------------------------------------------------------.
}
}
} I am working with archival specimens of formalin-fixed paraffin embedded
} tissues and I am getting tremendous autofluorescence. I looked at tissue
} sections after the sections have been de-waxed using either FITC or
} rhodamine filter sets and red cells and connective tissues are brightly
} fluorescent. Is there a way of suppressing the autofluorescence and still
} retain reactivity of tissue to antibodies? I would appreciate any comments
} or suggestions.
}
} *******************************************************
} Corazon D. Bucana, Ph.D.
} Department of Cancer Biology
} U.T. M.D. Anderson Cancer Center
} 1515 Holcombe Blvd. Box 173
} Houston, Texas 77030
} Phone: (713) 792-8106
} FAX: (713) 792-8747
} Email:bucana-at-audumla.mdacc.tmc.edu
} FAX: (713) 792-8747
}
}
}





From: Diana van Driel :      dianavd-at-eye.usyd.edu.au
Date: Fri, 11 Feb 2000 10:01:58 +1100
Subject: Re: New Gold Enhancement Reagents

Contents Retrieved from Microscopy Listserver Archives
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At 8:51 AM +0100 4/2/00, Alexander Mironov Jr. wrote:
} I have tried GoldEnhance for preembedding protocol to label intracellular
} structures. It is marvelous. Buy it and use it - you will not be
} unsatisfied. I have no any interest in Nanoprobes, I just like these dense
} gold particles.
} Practically all they claim is true:
} - light insensitive
} - no self-nucleation
} - do not react with buffer ions
} - is not dissolved by uranil and osmium (I have treat for 1 h)

I have been using BioCell's silver enhancement kit for years and
particularly like it for the light insensitivity - one can monitor
the reaction under the light microscope. Is it possible to do this
with the GoldEnhance as well?

GoldEnhance sounds perfect - has anyone had any problems with it?

Diana



Diana van Driel
Department of Clinical Ophthalmology
Sydney University
GPO Box 4337
Sydney NSW
AUSTRALIA 2001

Phone 61 2 938 27278/27395
Mob 0412 165 075
Fax 61 2 938 27318




From: Paul Webster :      pwebster-at-hei.org
Date: 10 Feb 00 20:26:38 -0800
Subject: Re:FYI: Congress to allow email charges -Hoax?

Contents Retrieved from Microscopy Listserver Archives
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Reply to: Re:FYI: Congress to allow email charges -Hoax?
I too had my suspicions. It looks very much like the "Internet Access Tax" letter from a few years ago. See: {http://urbanlegends.miningco.com/culture/urbanlegends/library/weekly/aa010798.htm} for details.

There are many sites documenting this sort of stuff. {http://www.hoaxkill.com/} is an example .

Disclaimer: I have no affiliation to this site.

Regards,
Paul Webster

Paul Webster, Ph.D.
Associate Scientist & Director
Ahmanson Advanced Electron Microscopy & Imaging Center
House Ear Institute
2100 West Third St.
Los Angeles, CA 90057

Phone: (213) 273-8026
Fax: (213) 413-6739
e-mail: pwebster-at-hei.org
http://www.hei.org/htm/aemi.htm





From: Nelson+ Conti :      NelsonC51-at-excite.com
Date: Thu, 10 Feb 2000 21:07:34 -0800 (PST)
Subject: Re:FYI: Congress to allow email charges -Hoax?

Contents Retrieved from Microscopy Listserver Archives
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On Thu, 10 Feb 2000 13:16:34 -0600, Damian 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
} -----------------------------------------------------------------------.
}
}
} Woody,
}
} I hate to add to the bandwidth clutter but these are very old hoaxes.
}
} Damian
}
}
} } Is this yet another hoax eating time & bandwidth by being posted without
} } confirmation (like the last 3-4 times I saw this message in the pas year
or
} } two), or is it real?
}
}
Damian's response is indeed correct -- see posted responses by
"David_Bell-at-millipore.com", "EvexAnalyt-at-aol.com", Scott Wight, Kalman Rubin,
Dr. Gary Gaugler, donald j. marsh, Roger Moretz, Laurie Wallin and Damian.
Being new to the MSA listserver, I'm very relieved that such notices (err,
BANDWIDTH-OCCUPYING, ANNOYING HOAXES) are determined to be false by very
astute readers. I fell for this, and, upon reading other people's notices, I
am upset that someone would suggest spreading such hoaxes around like the
classic junk chain mail activities done at school (or even college). Such
notices clog up the space needed for important issues regarding microscopy.
Anyway .. I'm responding merely to alert other GULLIBLE people (like myself)
not to believe such notices, as well as be thankful that this has been a
recurring event in the MSA listserver.
Nelson Conti





_______________________________________________________
Get 100% FREE Internet Access powered by Excite
Visit http://freeworld.excite.com





From: Alexander Mironov Jr. :      amironov-at-cmns.mnegri.it
Date: Fri, 11 Feb 2000 10:31:02 +0100 (MET)
Subject: Re: New Gold Enhancement Reagents

Contents Retrieved from Microscopy Listserver Archives
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Dear Diana,

IMHO, the main problem with GoldEnhance is some inhomogenity in particle
size but for my purposes it is irrelevant. Also the kit works good in my
hands at pH 7.4 but does not at pH 6.5 (as claimed Nanoprobes).

Sincerely,
Dr. Alexander A. Mironov Jr., MD, PhD
Unit of Morphology
Dept. of Cell Biology and Oncology
Consorzio Mario Negri Sud
Via Nazionale, S.Maria Imbaro (Ch)
66030 Italy

Tel. 0039-0872-570-332
Fax 0039-0872-570-412
E-mail: amironov-at-cmns.mnegri.it







From: GUIDO LND :      lueoend-at-zzmk.unizh.ch
Date: Fri, 11 Feb 2000 10:31:14 +0100
Subject: TEM Formvar Film Cast on Glass

Contents Retrieved from Microscopy Listserver Archives
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Dear readers,
could anyone give me some hints or tips how I can separate a formvarfilm
(0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't
help at all. It works better with non-washed slides, but then I get an
uneven and dirty surface of the film.
I appreciate every kind of support.
Best regards

Guido Luond
Institute of Oral Microbiology and
General Immunology
Univ. of Zurich
Plattenstr. 11
CH-8028 Zurich, Switzerland
E-mail: lueoend-at-zzmk.unizh.ch





From: richard.beanland-at-gecm.com
Date: Fri, 11 Feb 2000 09:48:48 +0000 (GMT)
Subject: Re: TEM: Sample preparation for InSb

Contents Retrieved from Microscopy Listserver Archives
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Carlos,
sounds like you may be having a problem with differential milling rates. Try milling over a restricted angle; you can use oscillation (if your ion mill is set up to do this) or make C-shaped Ta plate shields which you fix on the sample holding plates of your ion mill, with the sample at the centre of the 'C'. If you put your sample in the plates such that the interface is parallel to the opening of the shield, the average direction of the ion beam will be perpendicular to the interface. This will prevent the ion beam milling parallel to the glue line and stop the layers disappearing before the substrate. If you stick to angles below 10 degrees you will get better results, and of course use liquid nitrogen cooling and/or iodine. My apologies if this isn't very clear - it's hard to describe everything with just words! I can send you a picture of the sample holder I use if you like.

Best regards,

Richard Beanland

} ------------------------------------------------------------------------
} 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'm working with a InSb sample prepared with tripod polishing.
} Since the layer is quite thick, more than 3 micron plus a 2 micron buffer
} layer, I am experimenting some difficult to prepare a good sample for
} cross section observation. The final polishing is done in a Gatan ion
} milling set using a low angle. The problem with my samples is how to get a
} uniform thickness from the top of the sample to the substrate. Usually the
} top of the sample is milled much faster than near to the substrate. I
} can't mechanically polish the sample too much, since the sample is
} somewhat fragile (brittle). Even in old papers people report this kind of
} problem when preparing InSb samples for TEM using ion milling. Does
} someone else suffered from this kind of problem: the top part of a
} sample being milled away? Oh yes, when I polish the sample I try to make
} wedge perpendicular to the sample surface, so the thickness of the sample
} is the same from the InSb layer to the substrate after the mechanical
} polishing. There is some correlation between the ion milling angle/energy
} with this effect? The Gatan epoxy I use to glue a Si piece on top of the
} sample should have some influence (charging)? Thanks.
}
} Kazuo
}
} o-------------------------------------------------------o
} | Carlos Kazuo Inoki |
} | Department of Physics - University at Albany |
} | 1400 Washington Ave.- Albany - NY - 12222 |
} o-------------------------------------------------------o
}

==============================================================
Richard Beanland
Caswell Technology,
Caswell,
Towcester,
Northants NN12 8EQ

e-mail richard.beanland-at-gecm.com
Tel. +44 1327 356363
Fax. +44 1327 356398
==============================================================
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Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."








From: Barbara Foster :      mme-at-map.com
Date: Fri, 11 Feb 2000 07:50:18 -0500
Subject: Re: Color vs. monochrome cameras - a different approach

Contents Retrieved from Microscopy Listserver Archives
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David,

Is your end goal hi res imaging or ratioing? If the latter, I saw an
interesting technology at Cell Bio which might be of help: the PARISS
Spectral imaging system from LightForm. I saw very early versions of this
technology some years ago at PITTCON but it has really matured and offers
an interesting alternative. PARISS can acquire 240 spectra simultaneously,
full field or in a region of interest. It really crashes through the old
barriers imposed by filter cubes. Also, despite the radical differences in
intensity, it can acquire the transmitted light image simultaneously with
the fluorescence image and present the combined results with perfect
registration. The system retrofits to existing microscopes and is
moderately priced, considering all the accessories (2 cameras,
spectrometer, motorized stage, software). If you are interested, visit
their website at lightforminc.com or contact Jeremy Lerner (908-281-9098).

Caveat: MME has no financial interest in this product.

Hope this is helpful.

Best regards,
Barbara Foster
Consortium President
Microscopy/Microscopy Education ...Educating microscopists for greater
productivity.

125 Paridon Street Suite 102 Springfield, MA 01118
PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Visit our web site {http://www.MME-Microscopy.com/education}



At 10:23 AM 2/10/00 -0500, David Knecht wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America




From: L. D. Marks :      ldm-at-apollo.numis.nwu.edu
Date: Fri, 11 Feb 2000 07:35:08 -0600 (CST )
Subject: Re: coherence and spot size

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Sorry, I think a small correction/clarification is requires. This is ONLY
for an incoherently filled condensor aperture, i.e. a large beam with C2
fully focussed. This approximation is not true with the newer microscopes!

On Thu, 10 Feb 2000, Jonathan Barnard wrote:

} Born & Wolf (section 10.4.2)
} "Hence if the linear dimensions of the source and the distance between P1
} and P2 (points in the imaging plane) are small compared to the distance of
} these points from the source, the degree of coherence, |mu_12| is equal to
} the absolute value of the normalized Fourier transform of the intensity
} function of the source"
}

++++++++++++++++++++++++++++++++++++++++++++++++
Laurence Marks
Department of Materials Science and Engineering
Northwestern University
fax: (847) 491-7820
mailto:l-marks-at-nwu.edu
http://www.numis.nwu.edu
++++++++++++++++++++++++++++++++++++++++++++++++





From: Hans Dijkstra :      hans.dijkstra-at-edax.nl
Date: Fri, 11 Feb 2000 14:51:03 +0100
Subject: RE: Be X-ray peaks, solid state physics, how about 2nd row elements

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Scott,

WDS spectra of Boron, Carbon, Nitrogen and Oxygen in many compounds have
been extensively recorded by Bastin and Heijligers in the 1980's and early
1990's. They report that peak shifts and peak shape alterations are clearly
present in Boron compounds. In the case of TiB crystals it was even found
that the crystal orientation influences the peak shape and position, i.e. if
you rotate the sample 90 degrees you get a different peak shape. For Carbon
the peak shapes are practically unaffected by the bonding with other
elements.

Please check "Quantitative Electron Probe Microanalysis of Boron in Binary
Borides" by Bastin and Heijligers, University of Technology Eindhoven, the
Netherlands, 1997, ISBN 90-3860-898-5.

In EDX the peak shifts are undetectable. Modern EDX systems give a FWHM of
around 60-65 eV for these very light elements, so a peak shift of a few eV
is basically undetectable. For many users this makes EDX the prefered
technique to quantify Boron compounds, since with WDS you need to record
either the full peak (tedious) or use fixed area-peak-fraction correction
methods. In EDX we can simple ignore peak shape and peak position changes.

Best regards,

Hans Dijkstra

Disclaimer: This is my opinion, and not necessarily the one from EDAX Inc.
-------------------------------------------------------------
EDAX Europe www.edax.com
Ringbaan Noord 103 Tel.: +31-13-5364000
P.O. Box 4144 Fax.: +31-13-5356279
5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl
the Netherlands
-------------------------------------------------------------

} -----Original Message-----
} From: Walck. Scott D. [mailto:walck-at-ppg.com]
} Sent: Thursday, February 10, 2000 7:51 PM
} To: 'Microscopy'
} Subject: RE: Be X-ray peaks, solid state physics, how about 2nd row
} elements
}
}
} ------------------------------------------------------------------------
} 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 that I know understand the reason why you get Be X-ray and why
} there are energy shifts when it is in different compounds. My next
question
} then is do the elements between B and F have measurable energy shifts
} depending on what material they are in? The key word here is measurable.
} The Ka1 and Ka2 peaks are due to transitions form the 2p1/2 and 2p3/2 (LII
} and LIII) to the 1s1/2 state. But these would be valence/conductance
bands
} for the pure elements. (Assuming you collect your X-rays from solid N2,
O2,
} or F2) The band structure would be different for different materials. For
} example, do you see any shifts between B2O3? and BN or C(diamond) and
} C(graphite) or TiC? Any microprobers following this thread?
}
} -Scott
}
} Scott D. Walck, Ph.D.
} PPG Industries, Inc.
} Glass Technology Center
} Guys Run Rd. (packages)
} P. O. Box 11472 (letters)
} Pittsburgh, PA 15238-0472
}
} Walck-at-PPG.com
}
} (412) 820-8651 (office)
} (412) 820-8161 (fax)
}





From: Massimo Catalano :      massimo.catalano-at-ime.le.cnr.it
Date: Fri, 11 Feb 2000 15:08:23 +0100
Subject: PIPS and milling damage. Second message.

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Listservers:

first of all I would like to thank of the people who spent part of their
time trying to share their knowledge with me, and also to the manufacturers
who of course replied my messages.

All messages contained very important suggestions, that I need to study and
evaluate, and I will contact personally the people that offered to share
their previous experience and/or to collaborate.

Our work on InGaAs/GaAs quantum wells, single and stacked quantum wires,
single and stacked quantum dots, consists of conventional TEM, high
resolution transmission electron microscopy and high spatial resolution
analytical electron microscopy, that we perform in close collaboration with
Arizona State University, using a FEG/STEM machine equipped with EELS and
EDX. One of the goals of the research is to evaluate compositional
fluctuations in the well and in the dots at nanoscale.

As everybody knows, the requirements from a TEM sample for imaging are
completely different from the requirements necessary to perform high
spatial resolution analytical work, were effects such as amorphisazion,
surface contamination are critical.

We have a pretty well estabilished specimen preparation technique, but
before publishing results into the scientific community, we want to be sure
that what we observe is what is in the sample (isn't this the main concern
of every microscopist?).

That's why all suggestions, opinions and critics are strongly appreciated.

Best regards

Massimo


Dr. Massimo Catalano
CNR-IME
Campus Universitario
Via Arnesano
73100 Lecce - ITALY
tel: + 39 0832 322362
fax: + 39 0832 325299
email: massimo.catalano-at-ime.le.cnr.it
http://www.ime.le.cnr.it
http://www.ime.le.cnr.it/sime/sime.htm






From: Tamara Howard :      howard-at-cshl.org
Date: Fri, 11 Feb 2000 09:13:34 -0500 (EST)
Subject: Re: TEM Formvar Film Cast on Glass

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


I don't wash the slides, just wipe the dry slide with a Kim-Wipe a few
times. Works like a charm, whatever the weather! (Very grey and nasty here
on "the island" today, by the way).

Tamara Howard
CSHL


On Fri, 11 Feb 2000, GUIDO L[ISO-8859-1] ND 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 readers,
} could anyone give me some hints or tips how I can separate a formvarfilm
} (0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't
} help at all. It works better with non-washed slides, but then I get an
} uneven and dirty surface of the film.
} I appreciate every kind of support.
} Best regards
}
} Guido Luond
} Institute of Oral Microbiology and
} General Immunology
} Univ. of Zurich
} Plattenstr. 11
} CH-8028 Zurich, Switzerland
} E-mail: lueoend-at-zzmk.unizh.ch
}
}
}





From: Tindall, Randy D. :      TindallR-at-missouri.edu
Date: Fri, 11 Feb 2000 08:27:09 -0600
Subject: Re: TEM: Formvar Film Cast on Glass

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Guido,

There is a product called "Victawet", which is available here in the U.S.
from various EM suppliers. It comes in a small vial that lasts forever and
is an off-white, waxy or soapy substance. We used to take our slides and
clean them very well with lint-free papers, then place them in a rack inside
a vacuum evaporator, with the side intended for the Formvar film facing a
tungsten basket connected to the electrodes. Then we place a piece of
Victawet about the size of a grain of rice in the basket, pump down the
system to high vacuum, and slowly heat the tungsten basket until it glows
dull red. (If you increase the current too quickly the little piece will
jump out of the basket.) The slides will begin to look fogged, as if
someone had breathed moisture onto them. You can stop at that point---you
only need a little.

Store these coated slides until needed, then take them as required and
polish them very carefully with lint-free paper. They should feel slippery.
Use these cleaned slides to dip into the Formvar solution and the film
should separate easily.

Please note that the type of slide and the humidity levels in the workplace
also have a large effect. I have worked in places where films would
separate from any slide, every day, with no Victawet coating. I have also
worked in places where we could only get Formvar films on occasion (who
knows why?), and then we made a bunch of them at once.

Hope this helps.

Randy

Randy Tindall
EM Specialist
Electron Microscopy Core Facility
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211
(573) 882-8304
(573) 884-5414 (fax)
email: tindallr-at-missouri.edu {mailto:tindallr-at-missouri.edu}
http://biotech.missouri.edu/emc {mailto:tindallr-at-missouri.edu}



Dear readers,

could anyone give me some hints or tips how I can separate a formvarfilm

(0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't

help at all. It works better with non-washed slides, but then I get an

uneven and dirty surface of the film.

I appreciate every kind of support.

Best regards

Guido Luond

Institute of Oral Microbiology and

General Immunology

Univ. of Zurich

Plattenstr. 11

CH-8028 Zurich, Switzerland

E-mail: lueoend-at-zzmk.unizh.ch


Randy Tindall
EM Specialist
Electron Microscopy Core Facility
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211
(573) 882-8304
(573) 884-5414 (fax)
email: tindallr-at-missouri.edu {mailto:tindallr-at-missouri.edu}
http://biotech.missouri.edu/emc {mailto:tindallr-at-missouri.edu}





From: mike.santanajr-at-amd.com
Date: Fri, 11 Feb 2000 08:48:14 -0600
Subject: RE: FYI: Congress to allow email charges -Hoax? (STOP)

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Greetings,

Hasn't anyone realized that the "hoax" email is a success even as a
failure. Everyone is responding so much about the email being a hoax that
we're using up band width and cluttering email accounts. We all know it's a
hoax..., let's leave it at that.....


Regards,

Mike Santana, Jr.
Fab25 Product Engineering
Advanced Micro Devices
Desk - (512) 602-4172
Pager - (512) 622-2494
email - mike.santanajr-at-amd.com

On Thursday, February 10, 2000 10:27 PM, Paul Webster
[SMTP:pwebster-at-hei.org] 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
} -----------------------------------------------------------------------.
}
}
} Reply to: Re:FYI: Congress to allow email charges -Hoax?
} I too had my suspicions. It looks very much like the "Internet Access
Tax" letter from a few years ago.
See: {http://urbanlegends.miningco.com/culture/urbanlegends/library/weekly/aa
010798.htm} for details.
}
} There are many sites documenting this sort of stuff.
{http://www.hoaxkill.com/} is an example .
}
} Disclaimer: I have no affiliation to this site.
}
} Regards,
} Paul Webster
}
} Paul Webster, Ph.D.
} Associate Scientist & Director
} Ahmanson Advanced Electron Microscopy & Imaging Center
} House Ear Institute
} 2100 West Third St.
} Los Angeles, CA 90057
}
} Phone: (213) 273-8026
} Fax: (213) 413-6739
} e-mail: pwebster-at-hei.org
} http://www.hei.org/htm/aemi.htm
}




From: Jess Ricote :      jricote-at-icmm.csic.es
Date: Fri, 11 Feb 2000 16:21:51 +0100
Subject: ion thinner: users of Bal-Tec RES 010

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear listmembers,

I would like to contact users of Bal-Tec RES 010 ion thinner to comment on
performance of this equipment. We have found over the years frequent
stability problems of the guns, which are not solved only with cleaning. We
are at the moment checking any possible source of unstability. We would
like advise from anyone who has had similar problems with this apparatus,
to know if we are missing something and what would be the best course of
action.

Thanks in advance

Jess Ricote
-------------------------------------------------------------
Dr. Jess Ricote
Departamento de Materiales Ferroelctricos
Instituto de Ciencia de Materiales de Madrid
Consejo Superior de Investigaciones Cientificas (CSIC)
Cantoblanco
28049 Madrid
SPAIN

Phone: ( + 34) 91 334 90 00 (Ext. 268)
Fax: ( + 34) 91 372 06 23
e-mail: jricote-at-icmm.csic.es
http://www.icmm.csic.es/mf/ferroesp.htm
-------------------------------------------------------------





From: Massimo Catalano :      massimo.catalano-at-ime.le.cnr.it
Date: Fri, 11 Feb 2000 16:22:30 +0100
Subject: PIPS and milling damage: Second message

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Listservers:

first of all I would like to thank of the people who spent part of their
time trying to share their knowledge with me, and also to the manufacturers
who of course replied my messages.

All messages contained very important suggestions, that I need to study and
evaluate, and I will contact personally the people that offered to share
their previous experience and/or to collaborate.

Our work on InGaAs/GaAs quantum wells, single and stacked quantum wires,
single and stacked quantum dots, consists of conventional TEM, high
resolution transmission electron microscopy and high spatial resolution
analytical electron microscopy, that we perform in close collaboration with
Arizona State University, using a FEG/STEM machine equipped with EELS and
EDX. One of the goals of the research is to evaluate compositional
fluctuations in the well and in the dots at nanoscale.

As everybody knows, the requirements from a TEM sample for imaging are
completely different from the requirements necessary to perform high
spatial resolution analytical work, were effects such as amorphisazion,
surface contamination are critical.

We have a pretty well estabilished specimen preparation technique, but
before publishing results into the scientific community, we want to be sure
that what we observe is what is in the sample (isn't this the main concern
of every microscopist?).

That's why all suggestions, opinions and critics are strongly appreciated.

Best regards

Massimo


Dr. Massimo Catalano
CNR-IME
Campus Universitario
Via Arnesano
73100 Lecce - ITALY
tel: + 39 0832 322362
fax: + 39 0832 325299
email: massimo.catalano-at-ime.le.cnr.it
http://www.ime.le.cnr.it
http://www.ime.le.cnr.it/sime/sime.htm






From: richard.beanland-at-gecm.com
Date: Fri, 11 Feb 2000 15:56:43 +0000 (GMT)
Subject: Re: U-shaped Ta for IBT

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Ah. Bit of a problem, that. I didn't mention how far away the shield is from the sample - about 3 or 4 mm - and that for low angles I use a wedge-shaped shield to deflect ions away from the specimen, rather than back on to it. Did you really try it that quickly? I only sent the e-mail a few hours ago! I did expect anyone who was going to have a go to ask me for a picture of my holder.
As to cleaning your sample, I think you can only ion mill it some more, with amended shields...

Richard


} Dear Richard,
}
} I did as you suggested in my sample preparation. However, the Ta was
} sputtered on the surface of the
} specimen. Could you suggest me how to avoid or how to clean the Ta on the
} specimen surface.
}
} Chengge Jiao
} H.H.Wills Physics Lab
} University of Bristol, U.K
} c.g.jiao-at-bristol.ac.uk

==============================================================
Richard Beanland
Caswell Technology,
Caswell,
Towcester,
Northants NN12 8EQ

e-mail richard.beanland-at-gecm.com
Tel. +44 1327 356363
Fax. +44 1327 356398
==============================================================
"The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone.
Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."








From: Walck. Scott D. :      walck-at-ppg.com
Date: Fri, 11 Feb 2000 11:16:35 -0500
Subject: RE: PIPS and milling damage: Second message

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


One of the benefits from the small angle cleavage technique on the III-V
compounds is that often a "step-like" sample is produced. When this
happens, you have a perfect sample for both High resolution and analytical
work. In the very thin steps, you can do HREM and EELS. In the thicker
steps, you can do EDS analysis.
-Scott


Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)



} -----Original Message-----
} From: Massimo Catalano [mailto:massimo.catalano-at-ime.le.cnr.it]
} Sent: Friday, February 11, 2000 10:23 AM
} To: Microscopy-at-sparc5.Microscopy.Com
} Subject: PIPS and milling damage: Second message
}
}
} --------------------------------------------------------------
} ----------
} 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 Listservers:
}
} first of all I would like to thank of the people who spent
} part of their
} time trying to share their knowledge with me, and also to the
} manufacturers
} who of course replied my messages.
}
} All messages contained very important suggestions, that I
} need to study and
} evaluate, and I will contact personally the people that
} offered to share
} their previous experience and/or to collaborate.
}
} Our work on InGaAs/GaAs quantum wells, single and stacked
} quantum wires,
} single and stacked quantum dots, consists of conventional TEM, high
} resolution transmission electron microscopy and high spatial
} resolution
} analytical electron microscopy, that we perform in close
} collaboration with
} Arizona State University, using a FEG/STEM machine equipped
} with EELS and
} EDX. One of the goals of the research is to evaluate compositional
} fluctuations in the well and in the dots at nanoscale.
}
} As everybody knows, the requirements from a TEM sample for
} imaging are
} completely different from the requirements necessary to perform high
} spatial resolution analytical work, were effects such as
} amorphisazion,
} surface contamination are critical.
}
} We have a pretty well estabilished specimen preparation
} technique, but
} before publishing results into the scientific community, we
} want to be sure
} that what we observe is what is in the sample (isn't this the
} main concern
} of every microscopist?).
}
} That's why all suggestions, opinions and critics are strongly
} appreciated.
}
} Best regards
}
} Massimo
}
}
} Dr. Massimo Catalano
} CNR-IME
} Campus Universitario
} Via Arnesano
} 73100 Lecce - ITALY
} tel: + 39 0832 322362
} fax: + 39 0832 325299
} email: massimo.catalano-at-ime.le.cnr.it
} http://www.ime.le.cnr.it
} http://www.ime.le.cnr.it/sime/sime.htm
}
}
}




From: Heather A Owen :      owenha-at-csd.uwm.edu
Date: Fri, 11 Feb 2000 11:20:25 -0600 (CST)
Subject: Re: TEM Formvar Film Cast on Glass

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


I've always had good luck getting the film off by polishing the slide
first with Ross Optical Lens Tissue. My major professor taught me this,
and his explaination was that it has silicone in it. Once you have cast
the film onto the slide and allowed it to dry, cut through the film at the
edges and bottom of the slide by running a different slide along them.
Then breathe on the film and lower it into the water dish.

Good luck,
Heather Owen

p.s. I have no affiliation with the company that makes, or any of the EM
supply houses that sell Ross lens paper.

Heather A. Owen, Director
Electron Microscope Laboratory
Department of Biological Sciences
University of Wisconsin - Milwaukee
(414)229-6816






From: Mary Mager :      mager-at-interchange.ubc.ca
Date: Fri, 11 Feb 2000 09:41:58 -0800
Subject: RE: Be X-ray peaks, solid state physics, how about 2nd row

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Scott,
In the Microbeam Analysis 1985, G.F. Bastin and H.J.M. Heijligers have the
first article, entitled: "Quantitative Electtron-Probe Microanalysis of Very
Light Elements" (page 1). They have several articles in that and succeeding
years in which they try to analyse every element and compound in the B to F
range material that they can get to sit still under the EPMA beam. They did
a very valuable set of studies that is worth looking into if you want to do
any very light element analysis. They cover peak shifts, peak shape changes,
even crystallographic direction sensitivities.
At 01:51 PM 2/10/00 -0500, you wrote:

}
} I think that I know understand the reason why you get Be X-ray and why
} there are energy shifts when it is in different compounds. My next question
} then is do the elements between B and F have measurable energy shifts
} depending on what material they are in? The key word here is measurable.
} The Ka1 and Ka2 peaks are due to transitions form the 2p1/2 and 2p3/2 (LII
} and LIII) to the 1s1/2 state. But these would be valence/conductance bands
} for the pure elements. (Assuming you collect your X-rays from solid N2, O2,
} or F2) The band structure would be different for different materials. For
} example, do you see any shifts between B2O3? and BN or C(diamond) and
} C(graphite) or TiC? Any microprobers following this thread?
}
} -Scott
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





From: Elaine Humphrey :      ech-at-unixg.ubc.ca
Date: Fri, 11 Feb 2000 10:12:05 -0800
Subject: Re: questions for MSA members microwave

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hello Casey/Marty
I cannot answer for the first question as we use Diotome knives and I have
no experience with the other.

But the answer to the second question is "Yes." At least for us it is. We
are a facility used by the Faculty of Medicine and Science so many and
varied samples come through here for electron, light, and confocal
microscopy. We have been working up some protocols for difficult specimens
such as nematodes, drosophila larvae and plant material. The power supply
allows control so that you don't have to do any other processing to the
specimens than toss them in the fixative. Since the fix and post fix enter
the specimens at low power ( {200 watts) without damaging the specimens
(full power is usually about 800 watts) we can fix in about 45 minutes for
something which would normally be in 24 hours. And we don't have to punch
holes in the specimen.

We have just been playing with the introduction of fluorescent dyes for
light microscopy and confocal in C. elegans, without the use of agents to
distrupt the membranes to get the dye in. So far we have got the timings
for beautiful staining of the tail region and partial staining of the
thicker sections. We still have to get it right but the initial experiments
are very positive.

No-one seems to know why the microwaves work, but the hypothesis is that at
low power, the cell membranes are "jiggled about" to let the fix/dye in
without distrupting them.

We are a very busy lab, and our experiments are constantly getting
interupted so it is taking longer than we'd like to pin down a finished
protocol.

I would be interested in receiving protocols from labs who have already got
it to work for their specimens. We are finding that every specimen is
different and the protcols have to be tweaked for each one.
Elaine

}
} A colleague would like your responses to these questions. I believe that I
} say something on the first question about a year ago and apologize for
} asking again.

} } Here's what I/we need to know.
} }
} } 1) Are Microstar diamond knives equal to Diatome. We have an offer that
} } will save us about $1200 on three knives ( one 3mm ultra with 45 degree
} } angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife).
} }
} }
} } 2) Is the power controller feature on the Pelco Microwave Tissue processor
} } (model #3451) worth it? (costs about $1300 additional compared to the
} } Pelco 3450 without the power controller). I plan to do immunolocalization
} } work, but am not sure this power controller feature is necessary or worth
} } the cost.
} }
} } Casey

Marty Reed


Dr. Elaine Humphrey
Biosciences Electron Microscopy Facility
University of British Columbia
6270 University Blvd, mail-stop Botany
Vancouver, BC
CANADA, V6T 1Z4
Phone: 604-822-3354
FAX: 604-822-6089
e-mail: ech-at-unixg.ubc.ca






From: Maria.Fazio-Zanakis-at-aventis.com
Date: Fri, 11 Feb 2000 12:17:37 -0600
Subject: TEM Formvar Film Cast on Glass

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Dear Guido,
I have learned long ago not to use films cast on slides. It is easier
and much cleaner to spread your films on the surface of water in a trough.
You can vary the thickness by the amount of drops you use and chose the area
you want to place your grids. This method gave me beautiful and sturdy
films.
Regards,
Maria

Maria Fazio-Zanakis
Bioimaging and Molecular Histology
Aventis Pharmaceuticals
1-908-231-3357
Fax: 1-908-231-3962
Email: Maria.Fazio-Zanakis-at-Aventis.com


-----Original Message-----
} From: GUIDO LND [mailto:lueoend-at-zzmk.unizh.ch]
Sent: Friday, February 11, 2000 4:31 AM
To: EM-Tips


Dear readers,
could anyone give me some hints or tips how I can separate a formvarfilm
(0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't
help at all. It works better with non-washed slides, but then I get an
uneven and dirty surface of the film.
I appreciate every kind of support.
Best regards

Guido Luond
Institute of Oral Microbiology and
General Immunology
Univ. of Zurich
Plattenstr. 11
CH-8028 Zurich, Switzerland
E-mail: lueoend-at-zzmk.unizh.ch





From: Paul Webster :      pwebster-at-mailhouse.hei.org
Date: 11 Feb 00 10:43:54 -0800
Subject: RE: TEM Formvar Film Cast on Glass

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From: Paul Webster :      pwebster-at-mailhouse.hei.org
Date: 11 Feb 00 10:43:54 -0800
Subject: RE: TEM Formvar Film Cast on Glass

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Reply to: RE: TEM Formvar Film Cast on Glass
Hi Guido,

Here is my recipe which has worked on four continents at altitudes up to 2,000m and in all weathers (humid to dry, cold to hot). Plus, it does not use those very unscientific body fluids I often see being used on this list!

Dissolve the formvar in chloroform as a concentration higher than will make the correct film thickness (film thickness will vary with the speed the glass is drawn out of the solution as well as with formvar concentration).

Take a glass slide (it is important to use precleaned 25x75 mm slides, VWR Scientific, Cat number 48312-002) and dip it in absolute ethanol. Immediately dry it with Ross lens tissue. If you wash it any other way or dry it with anything other than the Ross lens tissue, the film will not come off the glass.

Coat the slide immediately and let the film dry. Score the edges of the film so that it will easily detach from the glass and float it onto the water surface. Evaluate the film thickness and adjust the formvar concentration by adding chloroform to the solution.

I know there are many versions of this method that work. However, this is the only method I have found that is completely oblivious to the weather (still sunny in LA) and has been reproduced by many people.
in my experience, the formvar dissolved in choroform does not seem to have the short shelf life I read about recently. I have used the same solution for years, topping it up with chloroform to keep it at a concentration that will produce thin films.

Regards,

Paul Webster

Paul Webster, Ph.D
House Ear Institute
2100 West Third Street
Los Angeles, CA 90057
phone:213 273 8026
fax: 213 413 6739
e-mail: pwebster-at-hei.org
http://www.hei.org/htm/aemi.htm


GUIDO LND wrote:
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} -----------------------------------------------------------------------.
}
}
} Dear readers,
} could anyone give me some hints or tips how I can separate a formvarfilm
} (0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't
} help at all. It works better with non-washed slides, but then I get an
} uneven and dirty surface of the film.
} I appreciate every kind of support.
} Best regards
}
} Guido Luond
} Institute of Oral Microbiology and
} General Immunology
} Univ. of Zurich
} Plattenstr. 11
} CH-8028 Zurich, Switzerland
} E-mail: lueoend-at-zzmk.unizh.ch
}
}
}
}
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}
Paul Webster, Ph.D
House Ear Institute
2100 West Third Street
Los Angeles, CA 90057
phone:213 273 8026
fax: 213 413 6739
e-mail: pwebster-at-hei.org
http://www.hei.org/htm/aemi.htm





From: Brian McIntyre :      mcintyre-at-optics.rochester.edu
Date: Fri, 11 Feb 2000 14:01:04 -0500
Subject: metal pipe- inside surface

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hi all-

just got a request to view surface structure of the inside of a 12" metal
pipe. its too big for my chamber and they'd prefer a non-destructive
technique. will replication of the surface with acetate/acetone and SEM
work. i've never tried this before so any pointers would be welcome...

thx!
brian
----------------------------------------------
Brian McIntyre
Electron Microscopy Lab- River Campus
Univ of Rochester
Rochester, NY 14627
716-275-3058/4875




From: Walck. Scott D. :      walck-at-ppg.com
Date: Fri, 11 Feb 2000 16:55:15 -0500
Subject: Fluorescent dyes -Clueless in Pittsburgh

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Laurie,
Frieda Carson's book about histologic technique is the "gold standard for
histotechs right now. It is available at Amazon.com and Barnes and Noble's
website, and can be special ordered at the major bookstores, as it is
currently in print. (Unfortunately I am at home and do not have the actual
title in front of me)
Wanda Shotsberger
----- Original Message -----
} From: Laurie Wallin {lwallin-at-ucsd.edu}
To: {microscopy-at-sparc5.Microscopy.Com}
Sent: Thursday, February 10, 2000 11:03 AM


I was asked a couple of questions about fluorescent microscopy and I didn't
have a clue. Are all the dyes used with UV or are there other wavelengths
that can fluoresce them? Are the dyes used dangerous/hazardous? Are they
transparent to visible? Are they expensive? Can you get them in quantity?

-Clueless in Pittsburgh


Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)





From: Jeff Allbright :      jeff.allbright-at-usa.net
Date: Fri, 11 Feb 2000 16:04:05 -0600
Subject: looking for ETEC Company

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I knew a very good, reputable company in the Southern California region that
serviced ETEC SEMs. The name was Scan Service in California, Earl Weltmer
714 area code.

- Jeff

-----Original Message-----
} From: LAB. DE MICROSCOPIA ELECTRONICA - FI - UNER
[mailto:microsc-at-fi.uner.edu.ar]
Sent: Wednesday, February 09, 2000 5:10 AM
To: microscopy-at-sparc5.microscopy.com


Hello .....
we have an old ETEC Omniscan SEM, and we are searching for ETEC company or
any
people that can supply manuals ....

any help is welcome

best regards


===================================================
Fernando D. Balducci
Laboratorio de Microscopia Electrnica
Facultad de Ingeniera - Bioingenieria
Universidad Nacional de Entre Rios
Argentina
e-mail: microsc-at-fi.uner.edu.ar
tel: 54 43 975100
fax: 54 43 975077


===================================================






From: Chuck Butterick :      cbutte-at-ameripol.com
Date: 1/4/80 8:23 AM
Subject: questions for MSA members

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In regards to the diamond knives, both companies make an excellent
product and both provide good service. Because of price, the last 6
knives I've bought have been from Microstar (two well-used but dull
Diatome knives have been traded during these transactions) . All six
knives have been more than suitable for the intended purpose. I have
no commercial interest in either company.

Chuck Butterick
Engineered Carbons, Inc.


______________________________ Reply Separator _________________________________


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A colleague would like your responses to these questions. I believe that I
say something on the first question about a year ago and apologize for
asking again.


} Here's what I/we need to know.
}
} 1) Are Microstar diamond knives equal to Diatome. We have an offer that
} will save us about $1200 on three knives ( one 3mm ultra with 45 degree
} angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife).
}
}
} 2) Is the power controller feature on the Pelco Microwave Tissue processor
} (model #3451) worth it? (costs about $1300 additional compared to the
} Pelco 3450 without the power controller). I plan to do immunolocalization
} work, but am not sure this power controller feature is necessary or worth
} the cost.
}
} Casey
}
}
}
Thanks for your time
}
Marty Reed
Equipment Technician
Biology Department
Humboldt State University
Arcata CA 95521
707-826-3234
707-826-3201 FAX
mmr7001-at-axe.humboldt.edu







From: White, Woody N :      Woody.N.White-at-mcdermott.com
Date: 2/11/00 1:01 PM
Subject: metal pipe- inside surface

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Date sent: Fri, 11 Feb 2000 14:01:04 -0500
To: Microscopy-at-sparc5.Microscopy.Com
} From: Brian McIntyre {mcintyre-at-optics.rochester.edu}


Depends on the detail needed, but acetate replication should work for almost
all
situations. Of course, everything you see will be inside out... Often,
selecting inverse video helps one perceive thing properly. If the pipe is
dirty or corroded, you will likely pick up some deposit with the replica.
This
can be distracting, but can also be helpful if x-ray analysis is required...
Woody White
McDermott Technology
My place: http://www.geocities.com/capecanaveral/3722

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hi all-

just got a request to view surface structure of the inside of a 12" metal
pipe. its too big for my chamber and they'd prefer a non-destructive
technique. will replication of the surface with acetate/acetone and SEM
work. i've never tried this before so any pointers would be welcome...

thx!
brian
----------------------------------------------
Brian McIntyre
Electron Microscopy Lab- River Campus
Univ of Rochester
Rochester, NY 14627
716-275-3058/4875




From: Mandayam V. Parthasarathy :      mvp2-at-cornell.edu
Date: Fri, 11 Feb 2000 18:23:11 -0500
Subject: PGT IMIX system module

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If any one has a PGT IMIX X-ray Analysis/Imaging system 4-40 (SUN Sparc)
that is in storage, or not in use, I would be interested in acquiring its
Image Signal Processor ISP module (Part# MO328D-00).



*******************************************************************

M.V. Parthasarathy
Prof. of Plant Biology, Adjunct Prof. of Biomedical Sciences (Vet), &
Director, Cornell Integrated Microscopy Center (CIMC)
Section of Plant Biology
228 Plant Science Building
Cornell University, Ithaca, NY 14853
E-Mail: mvp2-at-cornell.edu
Plant Biology Office: 268 Emerson; Telephone: 607-255-1734
Plant Biology Fax: 607-255-5407
CIMC Office: C1 054 Vet. Med. Center; Telephone: 607-253-3803
CIMC Office Fax: 607-253-3803
CIMC web site: http://www.cimc.cornell.edu






From: A.P. Alves de Matos :      apmatos-at-ip.pt
Date: Fri, 11 Feb 2000 19:12:34 -0600
Subject: TEM LKB control Unit

Contents Retrieved from Microscopy Listserver Archives
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Dear Brian

Stop destroying your pipe. With our large chamber SEM we can analyse your
pipe up to a length of 35 inches and a diameter of 28 inches. Maximum weight
of the sample should not exceed 500 pounds.

Because of the positionable electron gun it is easy to observe the inside
surface.

Best regards
Martin Klein

Disclaimer: VisiTec is the manufacturer of large chamber SEMs. And we like
large samples.

----------------------------------------------------------------------------
---
VisiTec Microtechnik GmbH
Karl-Marx-Str. 14
D-23936 Grevesmuehlen
Germany
Fon: +49-3881-79049
Fax: +49-3881-79048

email: info-at-visitec-em.de
WWW: http://www.visitec-em.de

----- Original Message -----
} From: Brian McIntyre {mcintyre-at-optics.rochester.edu}
To: {Microscopy-at-sparc5.microscopy.com}
Sent: Friday, February 11, 2000 8:01 PM


I am in desperate need of a control unit for a LKB ultratome III, since
mine is beyond repair.
I would appreciate if anyone coud sell me an unused unit from old
apparatus, even if it does not work, I might
be able to repair mine with the pieces.
Please reply to the e-mail below

Thanks

Dr. A.P. Alves de Matos
Dental Medical School
Lisbon University
apmatos-at-ip.pt






From: Brian Robertson :      brian.robertson-at-materials.oxford.ac.uk
Date: Fri, 11 Feb 2000 19:20:10 -0600
Subject: ELECTRON MICROSCOPY MATERIALS SPECIALIST

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ELECTRON MICROSCOPY MATERIALS SPECIALIST
UNL Center for Materials Research and Analysis

Analyze/characterize materials using electron microscopy, materials
preparation and computer instrumentation. Supervise/train students,
faculty and visiting researchers utilizing these methods. Bachelor's
with major in physical science, engineering or related field plus three
years experience in the operation, repair or design of electron
microscopes or other scientific instrumentation. Master's preferred.
Must have excellent computer and interpersonal/communication skills.
Knowledge of x-ray diffraction, TEM/SEM principles and sample
preparation experience preferred. Excellent benefits. Submit cover
letter, resume and names, addresses and telephone numbers of three
professional references to Professor David J. Sellmyer, 112 Brace Lab,
Lincoln, NE 68588-0113. Review of resumes will begin March 1. Position
will remain open until a suitable candidate is found. UNL is committed
to AA/EEO and ADA/504. If you require accommodation, please call (402)
472-8762.








From: michael baxter :      mykkb-at-juno.com
Date: Fri, 11 Feb 2000 19:21:23 -0600
Subject: Re: TEM-Formvar on Glass Slides

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Try a different brand of slide. Corning is the only brand we use in our EM
Class. There must be different kinds of surfactants used on them. Some
other brands of slides just don't release the formvar.
We usually wipe them off thoroughly with a Kimwipe only. Dip the slide
into the formvar and let dry. But don't dry for too long. Try using a
diamond tipped (or carbide) pen to score a "rectangle" on the slide.
Slowly insert the slide at about a 30 degree angle into the water. Wait
until you see the formvar start to release before you continue inserting
into the water.
This method usually works in our lab and for students.

Mike Baxter
Lehman College
Bronx, NY






From: Ken Tiekotter :      tiekotte-at-up.edu
Date: Sat, 12 Feb 2000 01:13:04 -0800 (PST)
Subject: Re: wanted: microtome

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Dear Linda,
I have a DuPont Sorvall MT2B or a Porter Blum MT1 I would be interested in
selling if the price is right. If you are interested, let me know via my
email address: tiekotte-at-up.edu.
Cheers!
Ken
-----------
Ken Tiekotter
Dept. of Biology
The University of Portland
5000 N. Willamette Blvd.
Portland, OR 97303

On Mon, 27 Aug 1956, Linda Chicoine wrote:

} ------------------------------------------------------------------------
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} -----------------------------------------------------------------------.
}
}
} Does anyone in the Connecticut area have an old microtome they would
} like to sell? I would be interested in anything from an LKB to RMC to
} Reichert/ and Leica. Please contact me by email. Thanks. Linda
} Chicoine
}
}
}





From: Garber, Charles A. :      cgarber-at-2spi.com
Date: Sat, 12 Feb 2000 03:32:42 -0600
Subject: SEM pipe replication

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} To: MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com}
} Subject: SEM pipe replication
} Date: Sat, 12 Feb 2000 10:44:32 -0500
} From: "Garber, Charles A." {cgarber-at-2spi.com}
} X-Mailer: E-Mail Connection v3.1a
} Content-Length: 3106

}
} -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
}
} Brian McIntyre wrote:
} =========================================
} just got a request to view surface structure of the inside of a 12" metal
} pipe. its too big for my chamber and they'd prefer a non-destructive
} technique. will replication of the surface with acetate/acetone and SEM
} work. i've never tried this before so any pointers would be welcome...
} ==========================================
} You did not indicate how far into the pipe you want to do your looking,
but
} cellulose acetate is certainly one approach. The biggest problem you are
} going to have is to balance the need to do this all "nondestructively" vs.
} the need to see what might be underneath material covering the top surface

} (after all, it is the metal substrate that you really want to see). In
} other words, if the acetone should be a solvent for the contaminating
} material, (or if it lifts off metal oxide or other aspects of a scale) you
} will not be doing this very non-destructively. One technique is to make
} repetitive replicas on the same area, each additional one taking off more
of
} the contaminating material, in order to get to the bare metal surface.
But
} this might not be considered, by a court of law, to be nondestructive.
And
} if you are under an order to look at it nondestructively, this could
result
} in a problem with the court.
}
} When we have been faced with this dilemma, in order to first document what
} was there originally, we like to use our SPI "Wet Replica" kit, it is a
} silicone based system, and is described on our website. Certain dental
} replication systems such as "Sil-Flow" might work as well, but in general,
} for this kind of work, we believe our own kit is better. The silicone
resin
} in general won't remove surface contamination or scale. The drawback of
this
} technique is that after about 800X in the SEM you start to see "structure"
} from the replication system itself. A "plus" however is that you can make
a
} "replica of the replica", doing your SEM viewing on a positive, which does
} look just like the original inside surface.
}
} Once you have documented the state of the inside surface that way, you
} should be able to get permission to try the cellulose acetate replication
} method. There is additional information on the SPI Supplies website
relative
} to cellulose acetate replication methods.
}
} Disclaimer: SPI Supplies offers both cellulose acetate replicating tapes
} and sheets as well as the silicone based SPI "Wet Replica Kit" for use in
} electron microscopy applications.
}
} 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.spi.cc
} ########################
} ============================================
}
}
}
}






From: Dagmar Preusse :      Preusse-at-chemie.uni-bremen.de
Date: Sat, 12 Feb 2000 12:54:22 +0100
Subject: permanent staining of LR-White embedded root sections

Contents Retrieved from Microscopy Listserver Archives
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Hi,
I made an embedding of the endodermis of an iris root with LR-Wihte and
would like to make ultrathin sections and stain them. My problem is how
to stain the sections and how to receive permantly stained sections. Is
there anyone who has experiences with the staining of suberin in the 3rd
endodermis? Please inform me about the handling.
Thank you so much
Dagmar Preusse





From: Wright, Cheryl W :      CWright-at-Sikorsky.com
Date: Sat, 12 Feb 2000 08:25:19 -0500
Subject: Photography of Coated Replicas

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


I run a 1830 Amray S.E.M. and work with failed aircraft components. From
time to time I am asked to replicate a surface using the acetate tape &
acetone method. The coating I use is a combination of Au and Pd. My
problem is that when I have to stay in one particular area of the replica,
the beam leaves a nice little square embedded into the surface of the
replica. I try to rapidly focus in full field instead of partial field, and
use between 6 - 10Kv. Does anyone have any advise for me on how to avoid
the embedded square?

Thanks, Cheryl




From: Steve Chapman :      PROTRAIN-at-CompuServe.COM
Date: Sat, 12 Feb 2000 09:40:28 -0500
Subject: Photography of Coated Replicas

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hi

It is the total current that you put into the specimen that is causing the
material to erode.

If you use a smaller spot size, smaller final aperture or smaller emission
current you should be better off? I do not know how good your SEM is at
low kV but if it was me I would use 2kV on this type of specimen; it will
be very sensitive even though it is coated.

To run happily at 2kV you need a filament to cathode distance that will
give you at least 30uA with your normal bias settings. If you do a good
deal of work on this type of specimen I would be inclined to lift the anode
by 5mm also. This will make the gun more efficient and make the job
easier.

Good luck

Steve Chapman
Protrain for Electron Microscopy Courses World Wide
E-mail - protrain-at-emcourses.com
Web Site - www.emcourses.com




From: Gordon Couger :      gcouger-at-rfdata.net
Date: Sat, 12 Feb 2000 13:32:41 -0600
Subject: [LM] replication

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


I have been trying to develop a styrene replica
fluid so I can use polorized light.So far the
styrene clumps ups in little islands insted of
making a nice film. I am using acetone as
a solvent

Has anyone had any luck with this?

Gordon

Gordon Couger gcouger-at-couger.com

Stillwater, OK www.couger.com/gcouger
405 624-2855 GMT -6:00






From: Mark Sanders :      msanders-at-biosci.cbs.umn.edu
Date: Sun, 13 Feb 2000 01:02:53 -0600
Subject: Re: questions for MSA members

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


{fontfamily} {param} Monaco {/param} Marty,

I can address #2 with some confidence.  We have been using the
Pelco

Microwave Tissue processor (model #3451) for almost 2 years with the
power

controller and have found it essential for optimizing the procedures we
use

for antigen retrieval and immunolocalization at both the light and EM
level.

By being able to control the power you have much better control of the

"microwave effect" relative to heating. I would be happy to discuss
the

details with you directly.

Cheers,

Mark

****************************************************

Mark A. Sanders
           University
of Minnesota

Program Director
          Twin Cities
Campus

Imaging Center
            St.
Paul, MN  55108

23-35 Snyder Hall
         ph:
 612-624-3454

1475 Gortner Ave.
         fax:
612-624-1799

{/fontfamily} http://biosci.umn.edu/imagingcenter {fontfamily} {param} Monaco {/param}

President: Minnesota Microscopy Society

{/fontfamily} http://resolution.umn.edu/MMS/ {fontfamily} {param} Monaco {/param}



----------

} From: Marty Reed { {mmr7001-at-humboldt.edu}

} To: Microscopy-at-sparc5.Microscopy.Com

} Subject: questions for MSA members

} Date: Fri, Jan 4, 1980, 11:23 AM

}


}
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America

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ListServer-at-MSA.Microscopy.Com

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}

}

} A colleague would like your responses to these questions.  I
believe that I

} say something on the first question about a year ago and apologize
for

} asking again.

}

}

} }        Here's what I/we need to
know.

} }

} } 1)  Are Microstar diamond knives equal to Diatome.  We have
an offer that

} } will save us about $1200 on three knives ( one 3mm ultra with 45
degree

} } angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife).

} }

} }

} } 2) Is the power controller feature on the Pelco Microwave Tissue
processor

} } (model #3451) worth it?  (costs about $1300 additional compared
to the

} } Pelco 3450 without the power controller).  I plan to do
immunolocalization

} } work, but am not sure this power controller feature is necessary or
worth

} } the cost.

} }

} } Casey

} }

} }

} }

} Thanks for your time

} }

} Marty Reed

} Equipment Technician

} Biology Department

} Humboldt State University

} Arcata CA 95521

} 707-826-3234

} 707-826-3201 FAX

} mmr7001-at-axe.humboldt.edu

}

}

{/fontfamily} Cheers, Mark
**************************************************** Mark A. Sanders
           University
of Minnesota Program Director
          Twin Cities
Campus Imaging Center
            St.
Paul, MN  55108 23-35 Snyder Hall
         ph:
 612-624-3454 1475 Gortner Ave.
         fax: 612-624-1799
http://biosci.umn.edu/imagingcenter President: Minnesota Microscopy
Society http://resolution.umn.edu/MMS/



{/x-rich}



From: Stuart McKernan :      mckernan-at-cems.umn.edu
Date: Sun, 13 Feb 2000 12:02:10 -0600 (CST)
Subject: Microscopy and Microanalysis 2000

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------- Start of forwarded message -------

Rules, FAQ Docs - Microscopy ListServer
To: NewSub-at-sparc5.Microscopy.Com
} From: "Nestor J. Zaluzec" {zaluzec-at-sparc5.Microscopy.Com}

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To: NewSub-at-MSA.Microscopy.Com
} From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com}


Here is a final reminder and IMPORTANT CORRECTION for the Call for Papers for
Microscopy and Microanalysis 2000.

The title for symposium #05 in the Call for Papers was listed incorrectly. The
correct title should be "Phase Transitions" (If you check the symposium titles
on the electronic data submission forms it is listed correctly). X-ray
microanalysis papers should be directed to symposia 25 and 26.

The deadline for papers is Tuesday February 15, which is almost upon us.

Stuart McKernan (Program Chair)




__________________
Stuart McKernan stuartm-at-tc.umn.edu
Director Office:(612) 626-7594
IT Characterization Facility, University of Minnesota Desk:(612) 624-6009
100 Union Street S. E., Minneapolis, MN 55455 NEW-} Fax:(612) 625-5368





From: Mel Dickson :      m.dickson-at-unsw.edu.au
Date: Mon, 14 Feb 2000 12:29:53 +1100
Subject: metal pipe- inside surface

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


} X-Sender: mcintyre-at-optics.rochester.edu
} Date: Fri, 11 Feb 2000 14:01:04 -0500
} To: Microscopy-at-sparc5.Microscopy.Com
} From: Brian McIntyre {mcintyre-at-optics.rochester.edu}
} Subject: metal pipe- inside surface
} Errors-to: Microscopy-request-at-sparc5.Microscopy.Com
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America


The best way to make replicas is to use dental casting compound.

It has the ability to replicate very fine detail and accepts even damp
surfaces.

We used "Kerr Extrude Wash" (two part polyvinylsiloxane impression
material) to replicate (damp) tuna fish skin.

Once cured, we made epoxy positive casts from the negative casts, then
mounted, gold coated etc. for regular SEM.

Your local dental supply house will have this.

Kerr USA is at 28200 Wick Road, Romulus MI 48174-2600


Dr. Mel Dickson,
Deputy Director, The Electron Microscope Unit,
Adjunct Associate Professor, School of Microbiology & Immunology
The University of New South Wales Sydney, Australia.
Phone +612 9385 6383 Fax +612 9385 6400




From: Robert H. Olley :      r.h.olley-at-reading.ac.uk
Date: Mon, 14 Feb 2000 09:24:08 +0000 (GMT Standard Time)
Subject: Re: [LM] replication

Contents Retrieved from Microscopy Listserver Archives
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Dear Malcolm,

You could also consider Epson Expression 1600 Pro. There is a review at the
PC Magazine site:

http://www.zdnet.com/pcmag/stories/firstlooks/0,6763,2430926,00.html

True optical resolution 1600x3200. Dynamic range 3.3
Although it is not from a scientific point of view the authors claim that
this is the best scanner for the price they've ever seen.

Good luck,

Rado

Disclaimer: I'm not related to Epson company in any way.

---------------------------------------------------------------------
Radostin Danev
Laboratory of Ultrastructure Research
National Institute for Physiological Sciences
Myodaiji-cho, Okazaki 444-8585, JAPAN
e-mail: rado-at-nips.ac.jp
---------------------------------------------------------------------
----- Original Message -----
} From: Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk}
To: Microscopy (MSA) {microscopy-at-sparc5.microscopy.com}
Sent: Thursday, February 10, 2000 8:26 PM



On Sat, 12 Feb 2000, Gordon Couger wrote:

} I have been trying to develop a styrene replica
} fluid so I can use polarized light.So far the
} styrene clumps up in little islands instead of
} making a nice film. I am using acetone as
} a solvent

Polystyrene is only partially soluble in acetone, so the low MW goes into
solution while the high MW forms clumps or whatever. BUTANONE (MEK)
should work fine, but you'll have to give it perhaps 4 times the drying
time you'd need with acetone. For samples that are sensitive to polar
solvents such as these, you can also use toluene, but that takes an even
longer drying time.

+------------------------------------------------------------------------+
| Robert H.Olley Phone: |
| J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 |
| University of Reading {University internal extension 7867 |
| Whiteknights Fax +44 (0) 118 9750203 |
| Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk |
| England URL: http://www.reading.ac.uk/~spsolley |
+------------------------------------------------------------------------+






From: Robert H. Olley :      r.h.olley-at-reading.ac.uk
Date: Mon, 14 Feb 2000 09:35:52 +0000 (GMT Standard Time)
Subject: Re: Photography of Coated Replicas

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html



On Sat, 12 Feb 2000, Wright, Cheryl W wrote:

} I run a 1830 Amray S.E.M. and work with failed aircraft components. From
} time to time I am asked to replicate a surface using the acetate tape &
} acetone method. The coating I use is a combination of Au and Pd. My
} problem is that when I have to stay in one particular area of the replica,
} the beam leaves a nice little square embedded into the surface of the
} replica. I try to rapidly focus in full field instead of partial field, and
} use between 6 - 10Kv. Does anyone have any advise for me on how to avoid
} the embedded square?

Polystyrene MIGHT be a better material to use than cellulose acetate,
which tends to chemically self-destruct under electron beams: in fact,
when there is acetate remaining on replicas for TEM I sometimes burn it
off SLOWLY with the electron beam (too fast and you leave carbonaceous
stuff). The aromatic nature of PS also allows it to take up more energy.

You could replicate with PS, as follows:

Make a solution of PS in butanone (MEK) in a glass Petri dish. Allow to
evaporate slowly (if you can fume it off from a closed dish in an oven at
about 50^C, that's better). This film can then either be cast onto the
component with butanone, or you can melt-form it above the glass
transition of PS, say about 150^C. Don't use PS windows directly from
envelopes, because the films contain a lot of built-in-strain.

I'm not sure of it's performance in SEM, because I've used it for TEM
replication of acetone-sensitive specimens.

+------------------------------------------------------------------------+
| Robert H.Olley Phone: |
| J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 |
| University of Reading {University internal extension 7867 |
| Whiteknights Fax +44 (0) 118 9750203 |
| Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk |
| England URL: http://www.reading.ac.uk/~spsolley |
+------------------------------------------------------------------------+






From: MARK GERMANI :      mmrinc-at-wwa.com
Date: Mon, 14 Feb 00 08:15:48 -0600
Subject: Pittcon Short Course

Contents Retrieved from Microscopy Listserver Archives
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} From: "Walck. Scott D." {walck-at-ppg.com}

------------------------------------------------------------------------
The Microscopy ListServer -- Sponsor: The Microscopy Society of America




So your companys SEM operator just retired. You have used light microscopes for years and they have asked you to run the SEM also. So now what do you do? The SEM manuals are long gone and do you really want to crack open that 800 page SEM/x-ray microanalysis textbook?

Dont worry, consider taking Electron Microscopy and Microanalysis a one-day short course to be held on Sunday, March 12, at PITTCON 2000 in New Orleans. The course is an introduction to SEM, TEM and x-ray microanalysis and is designed for analytical chemists and others who need to use these techniques for industrial problem-solving and product research and development.

For further information and online registration visit www.pittcon.org (Course #2029) or contact me offline.

Mark S. Germani, Ph.D.
MicroMaterials Research, Inc.
136 Shore Drive
Suite 200
Burr Ridge, IL 60521

(630) 325-8170
(630) 325-8178 (fax)

mmrinc-at-wwa.com





From: Patton, David :      David.Patton-at-uwe.ac.uk
Date: Mon, 14 Feb 2000 14:46:36 +0000 (GMT Standard Time)
Subject: Re: TEM LKB control Unit

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Electron Optical Services Ltd. make a replacement control
box. Their address is given below.

Dave



52 Higher Road,
Urmston,
Manchester,
M41 9AP
England
UK



Dave
On Fri, 11 Feb 2000 19:12:34 -0600 "A.P. Alves de Matos"
{apmatos-at-ip.pt} wrote:

} ------------------------------------------------------------------------
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}
}
} I am in desperate need of a control unit for a LKB ultratome III, since
} mine is beyond repair.
} I would appreciate if anyone coud sell me an unused unit from old
} apparatus, even if it does not work, I might
} be able to repair mine with the pieces.
} Please reply to the e-mail below
}
} Thanks
}
} Dr. A.P. Alves de Matos
} Dental Medical School
} Lisbon University
} apmatos-at-ip.pt
}
}
}

----------------------------------------
Patton, David
Email: David.Patton-at-uwe.ac.uk
"University of the West of England"





From: Orth99-at-aol.com
Date: Mon, 14 Feb 2000 10:40:43 EST
Subject: Looking at the inside of metal pipes...

Contents Retrieved from Microscopy Listserver Archives
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How often does this community need to look at the inside of metal pipes or
pipes in general?

And, what is the typical surface roughness you are dealing with, i.e. would
an instrument with 100 um to 200 um of Z range be able to handle it?

And, what is the typical x,y range, i.e. would an instrument with 2 mm image
field be acceptable?

Just wondering. Thanks for any input you are willing to share.


Carol Rabke, Ph.D.
Surface Metrology Marketing & Applications Consultant
716-425-0976




From: Yvonne Nmcov :      ynemcova-at-natur.cuni.cz
Date: Mon, 14 Feb 2000 16:53:33 +0100
Subject: embedding protocols

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Dear Microscopy Listserver members,
can someone provide me:
- a detailed protocol for embedding Chlamydomonas into the Lowicryl
Resin K4M (fixation media, desiccation solutions, times and
concentrations
that are suitable for Chlamydomonas)
- a fixation protocol for cryosectioning of Chlamydomonas

Thank you very much,
Yvonne Nemcova

--
*****************************************
Mgr. Yvonne Nemcova
Department of Botany, Charles University, Prague
Benatska 2. 128 01 Praha 2, Czech Republic
tel: 00-420-2-21953127, fax: 00-420-2-21953125
e-mail address: ynemcova-at-natur.cuni.cz
*****************************************






From: Virginie Serin :      serin-at-cemes.fr
Date: Mon, 14 Feb 2000 17:53:50 +0100
Subject: Re: EELS database, a new tool to play with

Contents Retrieved from Microscopy Listserver Archives
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Dear Scott,


Thank you for the information on the MSA/MAS format that you sent me
some weeks ago.


The MSA/MAS spectra file format is now implemented in the EELS and XAS
database, therefore the spectra can be downloaded in this format.
Please let me know if you have any other problems. Any more suggestions
?

Thank you for your help.


Best regards


Virginie









{center} -----------------------------------------------


{fontfamily} {param} New_York {/param} {bigger} {bigger} Virginie Serin

{/bigger} {/bigger} {/fontfamily} {fontfamily} {param} Geneva {/param} CEMES-CNRS,
29 Rue J. Marvig, BP 4347,

31 055 Toulouse Cedex 4, France

Tl: 33 (0)5 62 25 78 67, Fax : 33 (0)5 62 25 79 99,

e-mail: serin-at-cemes.fr

http://www.cemes.fr


{/fontfamily} EELS and X ray database : http://www.cemes.fr/~eelsdb/


-----------------------------------------------

{/center}


{/x-rich}



From: tea meulia :      meulia.1-at-osu.edu
Date: Mon, 14 Feb 2000 14:54:32 -0500 (EST)
Subject: Edwards Vacuum Evaporator

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We are having an Edwards Vacuum Evaporator model E12E2 with a control unit
EVM052 in excess. This instrument has not been used for several year and it
may need some maintanance work. If you are interested please contact Tea
Meulia (meulia.1-at-osu.edu) or Pat Ashbaugh (ashbaugh.11-at-osu.edu).


Tea Meulia



Tea Meulia
Research Scientist and Head
Molecular and Cellular Imaging Center
OSU/OARDC
1680 Madison Ave.
Wooster OH 44691

tel.: (330) 263'3828
fax: (330) 202'3563

http://www.oardc.ohio-state.edu/mcic






From: Gordon Couger :      gcouger-at-rfdata.net
Date: Mon, 14 Feb 2000 15:24:38 -0600
Subject: Re: [LM] replication

Contents Retrieved from Microscopy Listserver Archives
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Thanks to all that responded. I have a lot better handle on how to
do it. I will report back my results.

Thanks to all for providing a great source of infomation.

Gordon

Gordon Couger gcouger-at-couger.com

Stillwater, OK www.couger.com/gcouger
405 624-2855 GMT -6:00
----- Original Message -----
} From: "Robert H. Olley" {r.h.olley-at-reading.ac.uk}
} On Sat, 12 Feb 2000, Gordon Couger wrote:
}
} } I have been trying to develop a styrene replica
} } fluid so I can use polarized light.So far the
} } styrene clumps up in little islands instead of
} } making a nice film. I am using acetone as
} } a solvent
}
} Polystyrene is only partially soluble in acetone, so the low MW goes into
} solution while the high MW forms clumps or whatever. BUTANONE (MEK)
} should work fine, but you'll have to give it perhaps 4 times the drying
} time you'd need with acetone. For samples that are sensitive to polar
} solvents such as these, you can also use toluene, but that takes an even
} longer drying time.
}







From: Tom Reese :      treese-at-mbl.edu
Date: Mon, 14 Feb 2000 17:56:31 -0600
Subject: Stylus Pro 5000

Contents Retrieved from Microscopy Listserver Archives
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Epson has a new ink jet printer out-Stylus pro 5000-that is expensive
but they claim it is much better than the much cheaper models many of
us know and love. Would anyone know whether this is true of just
hype-hard to tell from the published specs...? Thanks...Tom Reese




From: Ken Tiekotter :      tiekotte-at-up.edu
Date: Tue, 15 Feb 2000 00:05:37 -0800 (PST)
Subject: Re: [LM] replication

Contents Retrieved from Microscopy Listserver Archives
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Dear Gordon,
Try xylene. It works quite nicely as a mounting medium of a differenting
refractive index from that of say, canada balsam or Permount. Email me if
you have questions. Good luck!
Cheers,
Ken

-------------
Ken Tiekotter
Dept. of Biology
The Univerisity of Portland
5000 N. Willamette Blvd.
Portland, OR 97303


On Sat, 12 Feb 2000, Gordon Couger wrote:

} ------------------------------------------------------------------------
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} -----------------------------------------------------------------------.
}
}
} I have been trying to develop a styrene replica
} fluid so I can use polorized light.So far the
} styrene clumps ups in little islands insted of
} making a nice film. I am using acetone as
} a solvent
}
} Has anyone had any luck with this?
}
} Gordon
}
} Gordon Couger gcouger-at-couger.com
}
} Stillwater, OK www.couger.com/gcouger
} 405 624-2855 GMT -6:00
}
}
}
}





From: Kuusisto, Ari :      Ari.Kuusisto-at-wallac.fi
Date: Tue, 15 Feb 2000 10:34:16 +0200
Subject: RE: autofluorescence quenching

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Corazon ,

You wrote
"I am working with archival specimens of formalin-fixed paraffin embedded
tissues and I am getting tremendous autofluorescence. I looked at tissue
sections after the sections have been de-waxed using either FITC or
rhodamine filter sets and red cells and connective tissues are brightly
fluorescent. Is there a way of suppressing the autofluorescence and still
retain reactivity of tissue to antibodies? I would appreciate any comments
or suggestions."

It is an unfortunate fact that most fixatives cause high autofluorescence
making the studies with conventional fluorescence microscope hard. The
disturbance of autofluorescence can be avoided by using time-resolved
fluorescence microscope and special labels (See the following articles:
Visl M, Sevus L, Kuusisto A, Harju R, and Soini
E: Time- resolved fluorescence microscopy: Elimination of autofluorescence
in tissue specimens for Image cytometry with fluorescent labelled probes.
Micron and Microscopica Acta 21; 3, (1990).

Sevus L, Kuusisto A, Visl M, Hemmil I., and
Soini E: Lanthanide
chelates as labels in in situ hybridization and
immunohistochemistry,
Micron and Microscopica Acta 21; 3, (1990).

Sevus L, Visl M, Syrjnen S, Sandberg M,
Kuusisto A, Harju R, Salo J, Hemmil I, Kojola H, and Soini E: Time-Resolved
Fluorescence Imaging of Europium Chelate Label in Immunohistochemistry and
In Situ Hybridization, Cytometry 13; 329, (1992).

Sevus L, Kuusisto A, Visl M, Hemmil I., Kojola
H., Roomans G. M. and Soini E.: Use of Fluorescent Europium Chelates as
Labels in Microscopy Allows Glutaraldefyde Fixation and Permanent Mounting
and Leads to Reduced Autofluorescence and Good Long-Time Stability,
Microscopy Research and Technique 27; 00, (1994).

I hope this information helps you.




Ari Kuusisto, Research Physicist

Phone: +358 2 267 8508 (direct)
+358 2 267 8111 (switchboard)
Fax: +358 2 267 8380
e-mail: Ari.Kuusisto-at-wallac.fi {mailto:Ari.Kuusisto-at-wallac.fi}
Address: Wallac Oy
PO Box 10
FIN-20101 Turku, Finland
Homepage: www.wallac.fi {http://www.wallac.fi}






From: Malcolm Haswell :      malcolm.haswell-at-sunderland.ac.uk
Date: Tue, 15 Feb 2000 14:02:33 +0000
Subject: Re: Stylus Pro 5000

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I saw the Epson web spec and it looks quite impressive - especially as
they seem to be quoting speeds for real picture printing, not 5%
coverage or text.

My only concern is that Epson now market two printers with "10 years
light fastness" (the Epson Stylus photo 870 - A4 and 1270 A3) so you
have to wonder which is better for e.m. prints.

Malcolm Haswell
Electron Microscopy
School of Sciences
Fleming Building
University of Sunderland
SUNDERLAND SR1 3SD
Tyne and Wear
UK

Tel (0191) 515 2872
e-mail: malcolm.haswell-at-sunderland.ac.uk

---------------------------------------------------------

Tom Reese wrote:
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
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}
} Epson has a new ink jet printer out-Stylus pro 5000-that is expensive
} but they claim it is much better than the much cheaper models many of
} us know and love. Would anyone know whether this is true of just
} hype-hard to tell from the published specs...? Thanks...Tom Reese




From: marienhoff.visitec-at-t-online.de (Marienhoff)
Date: Tue, 15 Feb 2000 15:10:14 +0100
Subject: Re: Looking at the inside of metal pipes...

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Dear Carol,

We were looking up to 70 mm deep inside cylinder bores and engine blocks
several times during the last months with our large-chamber SEM. Roughness
was typically very small (less than 10 m), but for small diameter tubes
the positioning system needs large travels, tilts, and rotary axes to inspect
every interesting area.

Disclaimer: Visitec manufactures the large-chamber SEM.

Best regards
Peter
___________________________
Dr. Peter Marienhoff
VisiTec Microtechnik GmbH
Karl-Marx-Str. 14
D-23936 Grevesmuehlen
Germany

Fon: +49-3881-790-47
Fax: +49-3881-790-48
email: pmarienhoff-at-visitec-em.de
http://www.visitec-em.de

"Orth99-at-aol.com"-at-sparc5.Microscopy.Com wrote:
} How often does this community need to look at the inside of metal pipes or
} pipes in general?
}
} And, what is the typical surface roughness you are dealing with, i.e. would
} an instrument with 100 um to 200 um of Z range be able to handle it?
}
} And, what is the typical x,y range, i.e. would an instrument with 2 mm image
} field be acceptable?
}
} Just wondering. Thanks for any input you are willing to share.
}
}
} Carol Rabke, Ph.D.
} Surface Metrology Marketing & Applications Consultant
} 716-425-0976
}





From: George Laing :      scisales-at-ngscorp.com
Date: Tue, 15 Feb 2000 10:12:15 -0500
Subject: Stylus Pro 5000

Contents Retrieved from Microscopy Listserver Archives
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The Epson 5000 is a printer/proofer using 6 color inkjet technology. The 6
color technology expands the available color gamut significantly. It is
available both with or without a Fiery RIP. The Fiery RIP is used for
Postscript processing of complex files from Quark, Pagemaker,
Illustrator,etc.
Without the Fiery box, an Epson software RIP is included. Print quality is
excellent although without the Fiery RIP, printing times can be
long,especially for 11x17 or larger prints. For more information see:
http://prographics.epson.com/products/stypro5000/index.html

Canon also has an excellent printer, the BJC-8500 that also uses 6 color
technology. The BJC-8500 also handles up to 13x19 paper with excellent
print quality. Canon utilizes a technology that optimizes ink drying and
water resistance. For more information see:
http://www.bjc8500.com/index2.html

Neither of these printers are speed demons in terms of page throughput, but
quality is excellent. I will be happy to answer any questions anyone may
have regarding output devices.

George Laing
National Graphic Supply

-----Original Message-----
} From: Tom Reese [mailto:treese-at-mbl.edu]
Sent: Monday, February 14, 2000 6:57 PM
To: Microscopy-at-sparc5.microscopy.com


Epson has a new ink jet printer out-Stylus pro 5000-that is expensive
but they claim it is much better than the much cheaper models many of
us know and love. Would anyone know whether this is true of just
hype-hard to tell from the published specs...? Thanks...Tom Reese






From: Dusevich, Vladimir :      DusevichV-at-umkc.edu
Date: Tue, 15 Feb 2000 09:25:24 -0600
Subject: RE: Photography of Coated Replicas

Contents Retrieved from Microscopy Listserver Archives
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I'd try, besides using low kV and beam current,
to increase substantially thickness of Au/Pd coating.

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

} -----Original Message-----
} From: Wright, Cheryl W [mailto:CWright-at-Sikorsky.com]
} Sent: Saturday, February 12, 2000 7:25 AM
} To: 'Microscopy-at-MSA.Microscopy.Com'
} Subject: Photography of Coated Replicas
}
}
} --------------------------------------------------------------
} ----------
} 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/MicroscopyListser} ver/FAQ.html
}
}
} --------------------------------------------------------------
} ---------.
}
}
} I run a 1830 Amray S.E.M. and work with failed aircraft
} components. From
} time to time I am asked to replicate a surface using the
} acetate tape &
} acetone method. The coating I use is a combination of Au and Pd. My
} problem is that when I have to stay in one particular area of
} the replica,
} the beam leaves a nice little square embedded into the surface of the
} replica. I try to rapidly focus in full field instead of
} partial field, and
} use between 6 - 10Kv. Does anyone have any advise for me on
} how to avoid
} the embedded square?
}
} Thanks, Cheryl
}




From: Pearl Martin :      image-at-optonline.net
Date: Tue, 15 Feb 2000 11:37:14 -0500
Subject: Job: Sr. Metallographic Tech SEM/X-ray

Contents Retrieved from Microscopy Listserver Archives
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I am a technical recruiter. Currently, I am working on a search for one
of my Northeast clients for a
Senior Metallographic Technician:


Will perform metallographic evaluation of coatings including work with
SEM (Scanning Electron
Microscopy) and X-ray. Will be responsible for the metallurgy
laboratory, including organization of
work, and operation and calibration of equipment. Will write technical
and engineering reports. Will
prepare projects for the metallurgy laboratory. Will maintain
communication with customers.

AAS/BS plus fours years experience in a metallography laboratory.

Resumes should be sent by email, mail or fax. For best results, please
send emails as attached
Word.docs - I have Word 97.

Pearl Martin
Image Associates Inc.
5254 Merrick Road
Massapequa, NY 11758
Phone (516)798-3993
Fax (516)797-8703
Email: pearl-at-jobspot.com





From: Furdanowicz, Waldemar :      rwafu-at-bsco.com
Date: Tue, 15 Feb 2000 12:19:14 -0500
Subject: RE: questions for MSA members (i.e. diamond knives)

Contents Retrieved from Microscopy Listserver Archives
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With regard to diamond knives, I think it depends on what you're cutting and
what quality of section you require.

We cut steel and cross sections of coatings (some very hard and brittle) on
steel, and the only knife that approaches satisfactory result is Diatome 35
degree, sharpened to extra fine edge. We traded our big old Microstar knife
for a 2nd Diatome, because it couldn't make good, electron transparent
sections.

On the other hand, if your material cuts as well with a glass knife as with
a diamond but you don't want to bother making your own glass knives, then go
ahead and save some dollars.

(In my view, the same reasoning applies to the brand of ultramicrotome to
use. My company is very frugal, and there is always strong pressure to save
a dollar here and there, but we found that only Reichert/Leica can
consistently produce good samples in our demanding application; and the
higher initial cost has been more than offset by savings in time and effort,
and by the quality of results. (I.e., often the ability to just get any kind
of a result).)

Valdemar Furdanowicz, Ph.D.
Research Labs - G165
Bethlehem Steel
Bethlehem PA 18016

No interest in Microstar, Diatome, Leica; and this not reflect an opinion or
position of my employer....etc.

-----Original Message-----
From: Chuck Butterick [mailto:cbutte-at-ameripol.com]
Sent: Friday, February 11, 2000 5:11 PM
To: Microscopy-at-sparc5.Microscopy.Com;
mmr7001-at-humboldt.edu
Subject: Re: questions for MSA members


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In regards to the diamond knives, both companies make
an excellent
product and both provide good service. Because of
price, the last 6
knives I've bought have been from Microstar (two
well-used but dull
Diatome knives have been traded during these
transactions) . All six
knives have been more than suitable for the intended
purpose. I have
no commercial interest in either company.

Chuck Butterick
Engineered Carbons, Inc.


______________________________ Reply Separator
_________________________________
Subject: questions for MSA members
Author: Marty Reed {mmr7001-at-humboldt.edu} at INTERNET-MAIL
Date: 1/4/80 8:23 AM



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A colleague would like your responses to these questions. I
believe that I
say something on the first question about a year ago and
apologize for
asking again.


} Here's what I/we need to know.
}
} 1) Are Microstar diamond knives equal to Diatome. We have
an offer that
} will save us about $1200 on three knives ( one 3mm ultra
with 45 degree
} angle, one 3mm ultra with 35 degree angle, one 10 mm histo
knife).
}
}
} 2) Is the power controller feature on the Pelco Microwave
Tissue processor
} (model #3451) worth it? (costs about $1300 additional
compared to the
} Pelco 3450 without the power controller). I plan to do
immunolocalization
} work, but am not sure this power controller feature is
necessary or worth
} the cost.
}
} Casey
}
}
}
Thanks for your time
}
Marty Reed
Equipment Technician
Biology Department
Humboldt State University
Arcata CA 95521
707-826-3234
707-826-3201 FAX
mmr7001-at-axe.humboldt.edu







From: Steve Barlow :      sbarlow-at-sunstroke.sdsu.edu
Date: Tue, 15 Feb 2000 09:47:15 -0800
Subject: deconvolution vendors

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We are considering the purchase of a separate deconvolution system to
augment our imaging lab's confocal system Any users or vendors with
information about such systems, please contact me offline.

thanks in advance

steve


Dr. Steven Barlow, Associate Director
EM Facility/Biology Department
San Diego State University
5500 Campanile Drive
San Diego CA 92182-4614
phone: (619)594-4523
fax: (619) 594-5676
email: sbarlow-at-sunstroke.sdsu.edu
website: http://www.sci.sdsu.edu/emfacility/

Chairman, Educational Outreach subcommittee
promoting access to microscopes
Microscopy Society of America http://www.msa.microscopy.com/






From: ERIC :      biology-at-ucla.edu
Date: Tue, 15 Feb 2000 10:37:36 -0800
Subject: Oven for curing blocks

Contents Retrieved from Microscopy Listserver Archives
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To the wealth of knowledge on the list,

We are in the market for a new oven for curing blocks.... Any
suggestions? This lab uses as a standard Eponate 12 as the epoxy resin of
choice....
The oven we currently have after being fixed several times does not see to
keep a constant temp.....






From: Wil Bigelow :      bigelow-at-engin.umich.edu
Date: Tue, 15 Feb 2000 15:11:33 -0400
Subject: Thermocouple gauges

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One of the members of this listserver group just asked me privately about
the fact that he had noticed that the reading of the thermocouple gauge on
his vacuum evaporator changed markedly when he jiggled the wires running
from the gauge tube to the gauge control unit. This is a matter that is
discussed in some detail on page 87 of my book 'Vacuum Methods in Electron
Microscopy' (see http://www.2spi.com/catalog/books/book48.html for a
description)

Actually, this is a rather common problem with thermocouple gauges which
all people using them should be aware of. It arises because the sensing
element in a thermocouple gauge is an arrangement of very fine wires that
form one or more thermocouple junctions. The output of these gauges is
thus the electrical potential produced by these junctions that is of the
order of only 10 or 15 millivolts DC. As a consequence, the signal detected
by the gauge controller can be strongly influenced by minor changes the
resistance of the electrical lines connecting the gauge to the controller.
Typically, unplugging one of the connectors in this line and reconnecting
it can change the resistance of the line sufficiently to cause a
significant change in the reading produced by the controller.

For this reason, considerable care must be exercised in installing and
maintaining thermocouple gauges if acceptable readings are to be obtained.
As a minimum, the same cables used in calibrating a gauge must subsequently
be used when it is put into operation, and the pins and sockets of the
plugs involved in the circuit must be kept clean and firmly connected.

Because of the low DC voltages that must be detected, the meters used in
the gauge circuit must be rather delicate and sensitive. We have
encountered situations when a static electric charge on the plastic window
of a meter affected the movement of the needle inside the meter enough to
prevent an accurate response. Coating the meter window with an anti-static
solution cured this situation.

As discussed in my book, Thermocouple gauges are rugged, inexpensive, and
very convenient for use in many applications, but they must be handled with
care if they are to give acceptable service.

Wilbur C. Bigelow, Prof. Emeritus
Materials Sci. & Engr., University of Michigan
Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-662-5237






From: Karen Kavanagh :      kavanagh-at-sfu.ca
Date: Tue, 15 Feb 2000 14:26:57 -0800 (PST)
Subject: TEM available Philips EM-300

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A working Philips EM 300 TEM is available for donation to
a non-profit organization.
Please contact Karen Kavanagh at
kavanagh-at-sfu.ca






From: Heather A Owen :      owenha-at-csd.uwm.edu
Date: Tue, 15 Feb 2000 18:02:57 -0600 (CST)
Subject: TEM - glutaraldehyde for perfusion

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Hello everyone,

A graduate student is getting ready for a rather large scale preparation
of rat brains for TEM using vascular perfusion. Luckily (since I'm a
botanist) her major professor has had experience with the procedure. He
doesn't know for sure whether he used EM or Biological grade
glutaraldehyde previously, but considering the large volume that will be
required, and the cost difference, I'm hoping that the Biological grade
will suffice.

I would appreciate advice from those of you that have experience with this
method of fixation.

Thank you in advance for any information.

Heather Owen

Heather A. Owen, Director
Electron Microscope Laboratory
Department of Biological Sciences
University of Wisconsin - Milwaukee
(414)229-6816






From: Jos Ulln Serrano :      jullan-at-mixcoac.upmx.mx
Date: Tue, 15 Feb 2000 18:05:34 -0600
Subject: epifluorescence

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Dear Colleagues:

I have attempted to purchase an epifluorescence nose from 'Carl Zeiss'
to fit the old "Universal" model scopes, but was dismayed to find out that
it is no longer available.
If anyone has one that is not being used and wishes to sell it, I would be
very interested.

Thanks

JOSE ULLAN SERRANO, Ph.D., M.D.

E-mail: jullan-at-mixcoac.upmx.mx

Departamento de ANATOMIA
Escuela de MEDICINA
Universidad Panamericana
c/ Donatello, 59
Col. Insurgentes-Mixcoac
03920 MEXICO, D.F.

http://www.mixcoac.upmx.mx
Telef. 55 98 33 02 / 55 63 13 43
FAX: 56 11 35 85




From: Dr. Gary Gaugler :      gary-at-gaugler.com
Date: Tue, 15 Feb 2000 20:05:09 -0600
Subject: Re: Thermocouple gauges

Contents Retrieved from Microscopy Listserver Archives
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Thermocouple gauges are more like gross indicators of vacuum. As a
resistive bridge, they are driven by constant current sources and the
bridge or sensor output is fed to a differential amplifier. This amplifier
has a gain and zero adjust potentiometer. If either of these are dirty
or defective, readings will be off. But if a thermocouple sensor is
exhibiting erratic readings, and it uses the standard vacuum tube socket
connector, I'd replace the detector right off the bat. They do eventually
fail. A $25 throw away isn't worth the hassle.

Most instruments, like SEMs, use the TC gauge to indicate that the mechanical
pump is working. Once the turbo pump spins up to } 80% RPM, the TC gauge
is rather useless. Better readings are from cold cathode gauges....which tend
not to fire very regularly. Hence, the optimum readings are from the ion pump
current.

At 01:11 PM 2/15/00 , you wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America





From: Sergey Ryazantsev :      sryazant-at-ucla.edu
Date: Tue, 15 Feb 2000 20:13:52 -0800
Subject: TEM - glutaraldehyde for perfusion

Contents Retrieved from Microscopy Listserver Archives
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Dear Heather

We perfuse 4% formaldehyde in 1x PBS only, than prepared the brain's slices
.3-.4 mm thick and fix them in a mixture 3-4% formaldehyde 2% GA (all - EM
grade) in 1x PBS. If you would like to add GA into perfusion solution - it
is only 0.1% - not a big deal. From another hand, I don't think Biological
grade GA will affects fixation quality. But, who knows, every time it is a
"game": if fixation is good - no problem, if - no, we will start to think
what's going on and "biological" grade may be a "case"... To avoid such
troubles I always used fresh GA and formaldehyde from non-opened ampoules.

Good luck, Sergey


} Date: Tue, 15 Feb 2000 18:02:57 -0600 (CST)
} From: Heather A Owen {owenha-at-csd.uwm.edu}
} Subject: TEM - glutaraldehyde for perfusion
} To: MSA Listserver {Microscopy-at-sparc5.Microscopy.Com}
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

Sergey Ryazantsev Ph. D.
Electron Microscopy
UCLA School of Medicine
Department of Biological Chemistry
Box 951737
Los Angeles, CA 90095-1737

Phone: (310) 825-1144
Pager: (310) 845-0248
FAX (departmental): (310) 206-5272
mailto:sryazant-at-ucla.edu
http://www.bol.ucla.edu/~sryazant






From: Sergey Ryazantsev :      sryazant-at-ucla.edu
Date: Tue, 15 Feb 2000 21:20:05 -0800
Subject: Thermocouple gauges

Contents Retrieved from Microscopy Listserver Archives
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Dear Wilbur

As for my knowledge (mostly from physic's course at 7th grade Russian
school) the thermocouple gauge has two "thermocouples": the probe itself
and reference thermocouple inside of gauge (separate "references" for each
type of probe, I believe). All together they generate enough response to
be easily measured. I agree that the major problem in your case is a bad
electrical contact in the connectors or even broken wire (it is well
possible, because some material uses for thermocouples are very fragile).
In my point of view, thermocouple gauges is very reliably and most accurate
devices and they do not need any special care.

I am using BARNANT-100 Thermocouple Gauge and REX-P96 Thermocouple
Controller with type "T" thermocouple sensors (copper/constantan, I
believe, but not sure) for many years without any problem. The sensor goes
into the vacuum chamber through regular 8-pin feed-thought. I was using
same as thermocouple material for screws to connect thermocouple's wires to
the feed-though wiring. For some reason, soldering does not work. I took
that screws from disassembled thermocouple connectors (looks like small
plugs) most suppliers sells for thermocouple probes (a dollar each). That
connectors should correspond to the thermocouple probe you are using.
Usually those connectors are color-coded. For instance connectors for my
type "T" probe was coded in blue. BARNANT-100 Thermocouple Gauge and
REX-P96 Controller performs self-calibration test every time you shut it ON
or disconnect/connect probe. During that calibration they "compensate"
some electrical abnormalities in the circuit like difference in the probe
length or as in my case the current deviation on the enter and end of the
feed-through. In theory, the "break" in thermocouple wiring on
feed-through should affect the gauge accuracy. In practice, I did not find
a difference between "solid" and with-feed-though probe up to 0.1oC
accuracy (which is sufficient for my needs). REX-P96 Thermocouple
Controller is just amazing. You may program complicated profiles for
cooling/heating cycles and it holds temperature pretty well. It has
special auto-calibration function to adjust heat rate depending from
parameters of your system. In practice it mean that your sample will not
overheated because of heater inertia. I am using that Controller in my
high-resolution shadowing device to control temperature from -150 to +50oC.

I have no interest in described products (only happy user) mostly because I
even don't know the names of real manufacturers. I remember it was
distributed by Cole-Parmer.

Sergey



} Date: Tue, 15 Feb 2000 15:11:33 -0400
} From: Wil Bigelow {bigelow-at-engin.umich.edu}
} Subject: Thermocouple gauges
} X-Sender: bigelow-at-srvr5.engin.umich.edu
} To: Microscopy Listserver {microscopy-at-sparc5.Microscopy.Com}
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

Sergey Ryazantsev Ph. D.
Electron Microscopy
UCLA School of Medicine
Department of Biological Chemistry
Box 951737
Los Angeles, CA 90095-1737

Phone: (310) 825-1144
Pager: (310) 845-0248
FAX (departmental): (310) 206-5272
mailto:sryazant-at-ucla.edu
http://www.bol.ucla.edu/~sryazant






From: Gordon Couger :      gcouger-at-rfdata.net
Date: Wed, 16 Feb 2000 01:53:56 -0600
Subject: Re: [LM] replication Thanks!!

Contents Retrieved from Microscopy Listserver Archives
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MEK worked great. I broke up the clear plastic from a
CDROM case and made a thick solution by adding MEK.
cleaned a slide and spread a drop of the solution on the
slide and put a celery leaf on it I covered the whole thing
with saran warp and gently pressed another slide on it
to get good contact. Then removed the slide and Saran
wrap.

After drying about 30 minutes I pealed the leaf off and
took a look under a Nomarski phase scope. It was
the best image I have seen on the scope. The Nomarski
colors were extremely strong and I could get three bands
of color in some features/artifacts. The effect of the styrene
on the polarized light were not as strong as I had hoped for
but they do increase the contrast and Nomarski colors. It
will make lovely photos. Now if my color CCD camera would
just get here. Monochrome just won't do it justice.

I has been a long time since I looked at a leaf but the
stoma showed up just like the books show and I remember.
Individual cells were very clearly visible.

The film is extremely fragile and probably needs a little
plastizer added and probably go to a go to a multi coat
system if the film needs to be pealed from a surface and
mounted. with the first coats being thin with little or no
plastizier and the following coats being thicker and
carrying more plastisizer.

My thanks to all the helped on this. I will probably try
other solvents but MEK does not set off my asthma
like xylene and toluene do. So I will probably put up
with the slower drying times.

I was very impressed with the detail that was replicated.

Thanks
Gordon

Gordon Couger gcouger-at-couger.com

Stillwater, OK www.couger.com/gcouger
405 624-2855 GMT -6:00




} From: Robert H. Olley [mailto:r.h.olley-at-reading.ac.uk]
} On Sat, 12 Feb 2000, Gordon Couger wrote:
}
} } I have been trying to develop a styrene replica
} } fluid so I can use polarized light.So far the
} } styrene clumps up in little islands instead of
} } making a nice film. I am using acetone as
} } a solvent
}
} Polystyrene is only partially soluble in acetone, so the low MW goes into
} solution while the high MW forms clumps or whatever. BUTANONE (MEK)
} should work fine, but you'll have to give it perhaps 4 times the drying
} time you'd need with acetone. For samples that are sensitive to polar
} solvents such as these, you can also use toluene, but that takes an even
} longer drying time.







From: Ken Tiekotter :      tiekotte-at-up.edu
Date: Wed, 16 Feb 2000 01:20:42 -0800 (PST)
Subject: Re: epifluorescence

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Dear Josi,
Try contacting John Oren at Vermont Optechs. I cannot tell you
immediately what his email or web site address is but his mail address is:
P.O. Box 69, Charlotte, VT 05445: Tel. (802) 425-2040; FAX. (802)
425-2074. Good luck!
-Ken


On Tue, 15 Feb 2000, Jos[ISO-8859-1] Ulln Serrano wrote:

} ------------------------------------------------------------------------
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} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
} -----------------------------------------------------------------------.
}
}
}
} Dear Colleagues:
}
} I have attempted to purchase an epifluorescence nose from 'Carl Zeiss'
} to fit the old "Universal" model scopes, but was dismayed to find out that
} it is no longer available.
} If anyone has one that is not being used and wishes to sell it, I would be
} very interested.
}
} Thanks
}
} JOSE ULLAN SERRANO, Ph.D., M.D.
}
} E-mail: jullan-at-mixcoac.upmx.mx
}
} Departamento de ANATOMIA
} Escuela de MEDICINA
} Universidad Panamericana
} c/ Donatello, 59
} Col. Insurgentes-Mixcoac
} 03920 MEXICO, D.F.
}
} http://www.mixcoac.upmx.mx
} Telef. 55 98 33 02 / 55 63 13 43
} FAX: 56 11 35 85
}
}





From: Alexander Mironov Jr. :      amironov-at-cmns.mnegri.it
Date: Wed, 16 Feb 2000 10:28:42 +0100 (MET)
Subject: TEM, biology: Diatome knives

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Dear All,

I have heard recently that Diatome invent some coating of their diamond
knives that significantly improve the quality of cryosections (they appear
more smooth, less compressed, practically without folds). Is it true or
just usual story to increase sales?

Sincerely,
Dr. Alexander A. Mironov Jr., MD, PhD
Unit of Morphology
Dept. of Cell Biology and Oncology
Consorzio Mario Negri Sud
Via Nazionale, S.Maria Imbaro (Ch)
66030 Italy

Tel. 0039-0872-570-332
Fax 0039-0872-570-412
E-mail: amironov-at-cmns.mnegri.it






From: David_Bell-at-Millipore.com
Date: Wed, 16 Feb 2000 08:25:55 -0500
Subject: Re: RJ Lee Instruments: Many Thanks!

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Hello again,

I want to thank everyone who took the time to reply to me. Your emails were
very informative and helpful, and I passed them along to my coworker.

Thanks again,

David Bell
Scientist
Electron Microscopy Lab
Millipore Corporation
80 Ashby Road
Bedford, MA 01730
(781) 533-2108






From: rlvaughn-at-unmc.edu
Date: Wed, 16 Feb 2000 08:48:39 -0600
Subject: TEM repair services

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Speaking of thermocouples, I need to replace (probably) the T1 and T2
thermocouples in our OLD Denton evaporator. Where, other than Denton, can
I look for these?
I also need service work done on our old Ultracut (the model before the "E"
series). I refuse to work with Leica. Is there anyone in the "real"
midwest area?





From: Bede Willenbring :      Bede.Willenbring-at-HBFuller.com
Date: Wed, 16 Feb 2000 09:20:53 -0600
Subject: Re: Thermocouple gauges

Contents Retrieved from Microscopy Listserver Archives
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I've also seen this type of behavior caused by a "cold" solder joint on one or more of the wires in the guage connector socket. Re-flowing the joints did wonders for stability.



============================================================
Bede Willenbring Phone: (651)236-5470
H.B. Fuller Company FAX: (651)236-5430
Research Chemist E-mail: Bede.Willenbring-at-HBFuller.com
P.O. Box 64683
St. Paul, MN 55164-0683



} } } Wil Bigelow {bigelow-at-engin.umich.edu} 02/15 1:11 PM } } }
------------------------------------------------------------------------
The Microscopy ListServer -- Sponsor: The Microscopy Society of America


One of the members of this listserver group just asked me privately about
the fact that he had noticed that the reading of the thermocouple gauge on
his vacuum evaporator changed markedly when he jiggled the wires running
from the gauge tube to the gauge control unit. This is a matter that is
discussed in some detail on page 87 of my book 'Vacuum Methods in Electron
Microscopy' (see http://www.2spi.com/catalog/books/book48.html for a
description)

Actually, this is a rather common problem with thermocouple gauges which
all people using them should be aware of. It arises because the sensing
element in a thermocouple gauge is an arrangement of very fine wires that
form one or more thermocouple junctions. The output of these gauges is
thus the electrical potential produced by these junctions that is of the
order of only 10 or 15 millivolts DC. As a consequence, the signal detected
by the gauge controller can be strongly influenced by minor changes the
resistance of the electrical lines connecting the gauge to the controller.
Typically, unplugging one of the connectors in this line and reconnecting
it can change the resistance of the line sufficiently to cause a
significant change in the reading produced by the controller.

For this reason, considerable care must be exercised in installing and
maintaining thermocouple gauges if acceptable readings are to be obtained.
As a minimum, the same cables used in calibrating a gauge must subsequently
be used when it is put into operation, and the pins and sockets of the
plugs involved in the circuit must be kept clean and firmly connected.

Because of the low DC voltages that must be detected, the meters used in
the gauge circuit must be rather delicate and sensitive. We have
encountered situations when a static electric charge on the plastic window
of a meter affected the movement of the needle inside the meter enough to
prevent an accurate response. Coating the meter window with an anti-static
solution cured this situation.

As discussed in my book, Thermocouple gauges are rugged, inexpensive, and
very convenient for use in many applications, but they must be handled with
care if they are to give acceptable service.

Wilbur C. Bigelow, Prof. Emeritus
Materials Sci. & Engr., University of Michigan
Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-662-5237








From: Geoff McAuliffe :      mcauliff-at-UMDNJ.EDU
Date: Wed, 16 Feb 2000 11:02:40 -0500
Subject: Re: TEM - glutaraldehyde for perfusion

Contents Retrieved from Microscopy Listserver Archives
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John Heckman wrote:

} Heather A Owen wrote:
}
} } Hello everyone,
} }
} } A graduate student is getting ready for a rather large scale preparation
} } of rat brains for TEM using vascular perfusion. Luckily (since I'm a
} } botanist) her major professor has had experience with the procedure. He
} } doesn't know for sure whether he used EM or Biological grade
} } glutaraldehyde previously, but considering the large volume that will be
} } required, and the cost difference, I'm hoping that the Biological grade
} } will suffice.
} }
} } Heather,
}
} It's been a while but, back when I did such things, I had bouts of
} non-uniform fixation when using biological grade glutaraldehyde solutions
} even in plant tissues. However, the solutions had been kept much longer than
} I'd now use and there are other preparation technique changes that confound
} the determination of causality. In the cases where I've perfused small
} animals, I've used EM grade GA since, as expensive as it is, it's cheaper
} than repeating the experiment.
}
} However, I did a full systemic perfusion. One random thought ( I haven't
} tried it): What's the vascular volume of a rat brain? If you just need just
} the brain, I'd think a little creative clamping (really small hemostats?) in
} the thorax should let you reduce the volume needed to make even a large
} experiment affordable with "the good stuff".
}
} cheers and good luck,
} John
}

Heather:

I agree with John on both counts. We clamp the aorta (and anything eles that
gets in the way) high in the abdomen to avoid perfusing organs we are not
interested in. Saves time and money. Also we tilt the animal once perfusion is
underway so that the head is lower.

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: Roger Moretz :      rcmoretz-at-excite.com
Date: Wed, 16 Feb 2000 08:21:17 -0800 (PST)
Subject: Re: TEM - glutaraldehyde for perfusion

Contents Retrieved from Microscopy Listserver Archives
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I have had a lot of years' experience doing vascular perfusion of the CNS of
rodents (rat and mouse). We have used the EM Sciences Biological Grade
glutaraldehyde, 50%, as our primary source for perfusion without problems
related to fixation. However, we have not used straight glutaraldehyde, but
found that 2% paraformaldehyde + 1% glutaraldehyde in 0.1M cacodylate buffer
at pH 7.0 to 7.2 gave consistent results with excellent preservation of
structures, morphology and cytoplasmic and nuclear components. We have used
cacodylate primarily because we use uranyl acetate stain either in the
dehydration/processing protocol and/or on the thin sections. Use of
phosphate buffers invariably results in precipitate formation, even when
used only on sections. Another reason for the paraform/glut/cacodylate
combination is that we have studied the leakage of tracers (e.g. HRP,
microperoxidase) across the blood brain barrier, and had to be able to move
into Tris buffer for DAB reactions, and phosphate buffer gave consistently
poorer results.

Hope this helps.

Roger Moretz
Dept. of Toxicology
BI Pharmaceuticals, Inc.

I have no personal or financial interest in EM Sciences--just a satisfied
user.


On Tue, 15 Feb 2000 18:02:57 -0600 (CST), Heather A Owen 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 everyone,
}
} A graduate student is getting ready for a rather large scale preparation
} of rat brains for TEM using vascular perfusion. Luckily (since I'm a
} botanist) her major professor has had experience with the procedure. He
} doesn't know for sure whether he used EM or Biological grade
} glutaraldehyde previously, but considering the large volume that will be
} required, and the cost difference, I'm hoping that the Biological grade
} will suffice.
}
} I would appreciate advice from those of you that have experience with
this
} method of fixation.
}
} Thank you in advance for any information.
}
} Heather Owen
}
} Heather A. Owen, Director
} Electron Microscope Laboratory
} Department of Biological Sciences
} University of Wisconsin - Milwaukee
} (414)229-6816
}
}
}


Roger C Moretz, Ph.D.





_______________________________________________________
Get 100% FREE Internet Access powered by Excite
Visit http://freeworld.excite.com





From: John Heckman :      heckman-at-pilot.msu.edu
Date: Wed, 16 Feb 2000 10:01:07 -0800
Subject: Re: TEM - glutaraldehyde for perfusion

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Heather A Owen 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 everyone,
}
} A graduate student is getting ready for a rather large scale preparation
} of rat brains for TEM using vascular perfusion. Luckily (since I'm a
} botanist) her major professor has had experience with the procedure. He
} doesn't know for sure whether he used EM or Biological grade
} glutaraldehyde previously, but considering the large volume that will be
} required, and the cost difference, I'm hoping that the Biological grade
} will suffice.
}
} I would appreciate advice from those of you that have experience with this
} method of fixation.
}
} Thank you in advance for any information.
}
} Heather Owen
}
} Heather A. Owen, Director
} Electron Microscope Laboratory
} Department of Biological Sciences
} University of Wisconsin - Milwaukee
} (414)229-6816

Heather,

It's been a while but, back when I did such things, I had bouts of
non-uniform fixation when using biological grade glutaraldehyde solutions
even in plant tissues. However, the solutions had been kept much longer than
I'd now use and there are other preparation technique changes that confound
the determination of causality. In the cases where I've perfused small
animals, I've used EM grade GA since, as expensive as it is, it's cheaper
than repeating the experiment.

However, I did a full systemic perfusion. One random thought ( I haven't
tried it): What's the vascular volume of a rat brain? If you just need just
the brain, I'd think a little creative clamping (really small hemostats?) in
the thorax should let you reduce the volume needed to make even a large
experiment affordable with "the good stuff".

cheers and good luck,
John

John Heckman
MSM Department
Michigan State University







From: Lesley Weston :      lesley-at-interchange.ubc.ca
Date: Wed, 16 Feb 2000 10:46:29 -0800 (PST)
Subject: Re: TEM - glutaraldehyde for perfusion

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


We use Biological grade to perfuse rats (whole body, we haven't tried clamping
bits off, though that might be a good idea since we're interested in just the
head). After the perfusion is finished, we remove the interesting bit, which is
huge by EM standards, and fix some more in Karnowski's. Then we go on to osmium,
and continue processing. We get acceptable EM pictures at high magnification,
even though our blocks are so large and processing times are therefore
extended. So I can recommend Biological grade glutaraldehyde. Hope this helps.

Lesley Weston.



On Tue, 15 Feb 2000, Heather A Owen 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 everyone,
}
} A graduate student is getting ready for a rather large scale preparation
} of rat brains for TEM using vascular perfusion. Luckily (since I'm a
} botanist) her major professor has had experience with the procedure. He
} doesn't know for sure whether he used EM or Biological grade
} glutaraldehyde previously, but considering the large volume that will be
} required, and the cost difference, I'm hoping that the Biological grade
} will suffice.
}
} I would appreciate advice from those of you that have experience with this
} method of fixation.
}
} Thank you in advance for any information.
}
} Heather Owen
}
} Heather A. Owen, Director
} Electron Microscope Laboratory
} Department of Biological Sciences
} University of Wisconsin - Milwaukee
} (414)229-6816
}
}
}
}






From: Larry Nittler :      nittler-at-lepvax.gsfc.nasa.gov
Date: Wed, 16 Feb 2000 17:09:21 -0500
Subject: TEM - Looking for ion mill

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Hello microscopy listserver-type people,

I am in the market for a used ion mill. Can anybody point me to one?

Thanks,
Larry

--
------------------------------------------------------------------------------
Larry R. Nittler Laboratory for Extraterrestrial Physics,
Code 691
Interstellar Dust Buster NASA Goddard Spaceflight Center
Greenbelt MD 20771
phone: 301-286-4572 fax:301-286-0212
nittler-at-lepvax.gsfc.nasa.gov http://www.ciw.edu/lrn/
------------------------------------------------------------------------------




From: RLevenson-at-cri-inc.com
Date: Wed, 16 Feb 2000 16:24:20 -0600
Subject: RE: autofluorescence quenching: spectral imaging can help

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} From: RLevenson-at-cri-inc.com Subject: RE: autofluorescence quenching:
spectral imaging can help

You asked about strategies for reducing autofluorescence.

In addition to various strategies which can include using near-IR dyes like
Cy5.5 and exciting far in the red, you can also treat tissues with sodium
borohydride or toluidine blue (these latter observations I pass on from
others--have not tried them myself).

I have recently had very promising results using spectral imaging to
discriminate autofluorescence from desired fluorescence. [Caution: I am
describing a product sold by my company]. CRI makes a liquid crystal
tunable filter (LCTF) that can be used to collect a stack of images at
different wavelengths. This yields a spectrum at every pixel of an image.
(A product with similar capabilities is sold by Applied Spectral Imaging,
but uses a different technology.) CRI's new LCTF, the FluoroSpec, uses
special optics to combine polarization states; along with other
improvements, it has more than twice the light throughput compared to
previous models.

The sample was formalin-fixed brain probed with a Cy2-labeled antibody to a
low-abundance protein. By eye it was very difficult to distinguish the Cy2
signal from the bright green to green-orange autofluorescence. Using a
spectral data set (15 images from 510 nm to 650 nm at 10 nm intervals) and
linear combination algorithms for pixel-unmixing, it was possible to
completely separate the Cy2 and autofluorescence images.

I would be glad to share these images with any interested parties.

Richard


Richard Levenson, MD
Technical Director, Biomedical Systems
Cambridge Research & Instrumentation (CRI), Inc.
80 Ashford St.
Boston, MA 02134

Ph: (617) 787-5700; Fax: (617) 787-4488
rlevenson-at-cri-inc.com www.cri-inc.com






From: chuck burilla :      burilact-at-ix.netcom.com
Date: Wed, 16 Feb 2000 18:29:41 -0500
Subject: Re: Stylus Pro 5000

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With respect to the Epson Stylus Pro 5000:

I just purchased one for printing images, x-ray maps, etc. obtained from my
new JEOL 8900 microprobe. Previous to this I was utilizing an Epson Photo
700 printer with my older Cameca MBX probe. I was very pleased with the 700
and for a few hundred dollars sure was worth it! Not a fast printer but gave
high quality photo-type prints.
After reading the 5000 specs I decided to get one for the new probe.
In a "nutshell" here are my observations:

I purchased just the 5000 printer (not the Fiery RIP server that interfaces
to it for network printing). Cost: around $3000. for the 5000 only.
The 5000 is a much more robust printer than the 700 type Epson models. It
can print up to 13"X19" formats. It is faster than the 700 series and the
ink cartridges and paper capacity are much greater.

To realistically compare the printers I ran a speed and print quality
comparison between my two models. I printed a full 8 1/2" X 11" full color
bleed consisting of 3 x-ray maps, a BSE image and an EDS spectrum, along
with some text. Printing on the same PC, under the same conditions the print
quality is about the same. (A little disappointing here). However the print
speed was a full 4 times faster on the 5000 as compared to the 700. Actual
print time with the 5000 was about 2.5 min. at 1440 DPI.
I found an excellent web site which compares various Epson Photo printers,
including the 5000 and 700 series, and demonstrates print quality issues
quite vividly:

http://www.tssphoto.com/sp/dg/news/dot_comp.html

All in all I believe the 5000 is a much higher caliber printer than the 700
series, but unfortunately the print quality is about the same.

One final note: the paper makes ALL the difference in the world with this
class of printers.
Email me if you need info and suggestions on paper issues...



Chuck Burilla
United Technologies Research Center
400 Silver Lane
East Hartford, CT 06108

(860) 610-7388
burilact-at-utrc.utc.com



-----Original Message-----
} From: Malcolm Haswell [mailto:malcolm.haswell-at-sunderland.ac.uk]
Sent: Tuesday, February 15, 2000 9:03 AM
To: Microscopy (MSA)


I saw the Epson web spec and it looks quite impressive - especially as
they seem to be quoting speeds for real picture printing, not 5%
coverage or text.

My only concern is that Epson now market two printers with "10 years
light fastness" (the Epson Stylus photo 870 - A4 and 1270 A3) so you
have to wonder which is better for e.m. prints.

Malcolm Haswell
Electron Microscopy
School of Sciences
Fleming Building
University of Sunderland
SUNDERLAND SR1 3SD
Tyne and Wear
UK

Tel (0191) 515 2872
e-mail: malcolm.haswell-at-sunderland.ac.uk

---------------------------------------------------------

Tom Reese 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
} -----------------------------------------------------------------------.
}
} Epson has a new ink jet printer out-Stylus pro 5000-that is expensive
} but they claim it is much better than the much cheaper models many of
} us know and love. Would anyone know whether this is true of just
} hype-hard to tell from the published specs...? Thanks...Tom Reese





From: Paul Webster :      pwebster-at-mailhouse.hei.org
Date: 16 Feb 00 15:54:53 -0800
Subject: General:Wall Posters

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------------------------------------------------------------------------
The Microscopy ListServer -- Sponsor: The Microscopy Society of America


Hello,

Does anyone know the name of a supplier of wall posters suitable for an EM lab. I am thinking of cut-out diagrams of TEM and SEM, history of EM (I think JEOL once had one like that), specimen preparation etc.
Regards,

Paul Webster, Ph.D
House Ear Institute
2100 West Third Street
Los Angeles, CA 90057
phone:213 273 8026
fax: 213 413 6739
e-mail: pwebster-at-hei.org
http://www.hei.org/htm/aemi.htm





From: oshel-at-terracom.net (Philip Oshel)
Date: Wed, 16 Feb 2000 19:57:42 -0600
Subject: Gordon Cougar

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My apologies to the list, but: could Gordon Cougar email me? I get a fatal
error when trying to email you:
The following addresses had permanent fatal errors -----
} {gcouger-at-rfdata.net}
[...]
} 550 {gcouger-at-rfdata.net} ... User unknown

Thanks!

Phil

*** Be famous! Send a Tech Tip or article to Microscopy Today! ***
Philip Oshel
Technical Editor, Microscopy Today
P.O. Box 620068
Middleton, WI 53562-0068
Voice: days (608) 263-4162, evening (608) 833-2885
Fax (608) 836-1969 (please make sure my name is on any fax)

Address for UPS, FedEx, or other couriers:
Dept. of Animal Health and Biomedical Sciences
University of Wisconsin
1656 Linden Drive
Madison, WI 53706-1581

oshel-at-terracom.net
or
peoshel-at-facstaff.wisc.edu






From: Steve D'Angelo :      steve-at-equiprx.net
Date: Thu, 17 Feb 2000 03:17:28 -0800
Subject: General:Wall Posters

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Paul,
My favorite has always been the DIATOME calendars.
Next are the Gatan calanders.
Dennis Kunkel has great original SEMs of diatomes etc, but I don't know if
he has any posters.
David Sharpe has the old standby insect SEMs.
The students at SanJoaquin Delta College have put together some good
calendars also.

} Date: 16 Feb 00 15:54:53 -0800
} From: Paul Webster {pwebster-at-mailhouse.hei.org}
} Subject: General:Wall Posters
} To: MSA listserver submission {Microscopy-at-sparc5.microscopy.com}
} X-Priority: 3
} MIME-Version: 1.0
} Reply-To: Paul Webster {pwebster-at-mailhouse.hei.org}
} X-MIME-Autoconverted: from quoted-printable to 8bit by
} ultra5.microscopy.com id RAA14282
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America


Very best regards,
Steve D'Angelo
Equipment Resurrection
650-738-0351

NOW at http://equiprx.net/






From: Ritchie Sims :      r.sims-at-auckland.ac.nz
Date: Thu, 17 Feb 2000 16:14:42 GMT+1200
Subject: DP fluid for JEOL 840

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html




Hello, Pumpers

Does anyone out there have direct experience of any benefit(s) gained
by the use of Santovac 5 in the diffusion pumps of a JEOL 840,
instead of the recommended Lion S?

And did they have to fiddle with either the heating elements or
heater supply voltages in order to acheive the benefits?

I know that Santovac is well thought of, but it's expensive, and my
experience of Lion S in an older JEOL (JXA-5A) has been very
positive. I think there's something in there about optimising the
pump design to a particular fluid.

As an economical alternative, mentioned in Wil Bigelow's book, has
anyone tried NEOVAC SY, marketed by Varian, in an 840?

Does anyone know what chemical family Lion S belongs to?

Perhaps someone from JEOL, or even from the Lion company might care
to reply.

cheers

Ritchie

Ritchie Sims Phone : 64 9 3737599 ext 7713
Department of Geology Fax : 64 9 3737435
The University of Auckland email : r.sims-at-auckland.ac.nz
Private Bag 92019
Auckland
New Zealand




From: Gordon Vrololjak :      gvrdolja-at-nature.Berkeley.EDU
Date: Thu, 17 Feb 2000 17:04:35 -0800 (PST)
Subject: Mikros Manual available

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hello,
I'm cleaning out some things in the laboratory and we have a Mikros Vacuum
Evaporator Manual C-20, 1964 here. We no longer have this evaporator, so
we no longer need the manual. If anyone would have need of it, please
email me personally and we'll send it out to you.

I'm amazed at the quality of this older manual - actual blue prints
included!

\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\
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





From: ard-at-ansto.gov.au (Arthur Day)
Date: Sat, 19 Feb 2000 09:58:59 +1100
Subject: Re: DP fluid for JEOL 840

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Ritchie,

There was a fairly extensive discussion about Dp oils on the listserver a
while ago. That should be in the archives somewhere.

}
} Does anyone out there have direct experience of any benefit(s) gained
} by the use of Santovac 5 in the diffusion pumps of a JEOL 840,
} instead of the recommended Lion S?
}

"Yes". We have been using Santovac 5 in our early 6400 machine for about 5
months now. The diffusion pumps appear to be the same as on an 840 and the
6400 had Lion S in it beforehand.

} And did they have to fiddle with either the heating elements or
} heater supply voltages in order to acheive the benefits?
}

No. The vacuum seems to be as good as ever without doing any of this. It
all looks very clean.

} I know that Santovac is well thought of, but it's expensive, and my
} experience of Lion S in an older JEOL (JXA-5A) has been very
} positive. I think there's something in there about optimising the
} pump design to a particular fluid.
}
} As an economical alternative, mentioned in Wil Bigelow's book, has
} anyone tried NEOVAC SY, marketed by Varian, in an 840?
}
} Does anyone know what chemical family Lion S belongs to?

That should all be in the archives somewhere, or perhaps on certain web
sites. Jim Darley and/or Will Bigelow had the answers.

}
} Perhaps someone from JEOL, or even from the Lion company might care
} to reply.
}

We were told that the Santovac 5 is more tolerant of the occasional gulp of
air.....so presumably it should last longer.

Cheers


Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457
Ansto Materials Division Fax: 61-2-9543-7179
PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au
Australia www: http://www.ansto.gov.au/






From: bozzola-at-siu.edu (John J. Bozzola)
Date: Fri, 18 Feb 2000 17:59:22 -0600
Subject: Photoshop 5.5 Thumbnail Preview in Macs

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


For imaging gurus:

I am using Photoshop 5.5 on a G3 Macintosh and can no longer get a preview
of the image when the file is selected in FILE } OPEN. What gives?

In previous versions (say 4.0 and earlier) when the file was selected, I
got a thumbnail preview of the image prior to opening it. This is very
convenient and I miss this capability.

Thanks,

John

####################################################################
John J. Bozzola, Ph.D., Director
Micro-Imaging and Analysis Center
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
####################################################################






From: steve :      halifa-at-globalnet.co.uk
Date: Sat, 19 Feb 2000 12:32:11 -0600
Subject: Re Santovac

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Santovac is a great oil to use in Jeol instruments. Backstreaming is
less and it will withstand a lot more abuse than Lion oil if the vac
system has a glitch. Just put a little less charge in than the normal Lion
oil. Regards Steve Parkins






From: Wright, Cheryl W :      CWright-at-Sikorsky.com
Date: Sat, 19 Feb 2000 12:32:33 -0600
Subject: Photography of Coated Replicas

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hi Everyone,

WOW, thanks soooo much for all the responses to my question. I have alot of
techniques to try so that should keep me busy for awhile. This is a very
informative site and I appreciate the help!!

Cheryl






From: Dean Abel :      dean-abel-at-uiowa.edu
Date: Sun, 20 Feb 2000 14:48:35 -0600
Subject: TEM Philips 300

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


The Department of Biological Sciences at the University of Iowa must find a
home for a Philips 300 Transmission Electron Microscope by June 2000. It
is in excellent condition and has been under service contract for nearly 30
years. We are remodeling our building and will no longer maintain support
for this instrument. Free to anyone willing to come and take it away (plus
Haskris chiller, carbon evaporator, film dessicator, etc.).

Please contact:
Dean Abel
Biological Sciences 138BB
University of Iowa
Iowa City IA 52242 USA
319-335-1070 (fax 319-335-1069)
dean-abel-at-uiowa.edu





From: Diana van Driel :      dianavd-at-eye.usyd.edu.au
Date: Mon, 21 Feb 2000 09:49:50 +1100
Subject: Photoshop 5.5 Thumbnail Preview in Macs

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear John,

Open the image in Photoshop and select SAVE AS. There are boxes to be
ticked, one of which is Mac thumbnail. My guess is that your problem
is here.

Diana



Diana van Driel
Department of Clinical Ophthalmology
Sydney University
GPO Box 4337
Sydney NSW
AUSTRALIA 2001

Phone 61 2 938 27278/27395
Mob 0412 165 075
Fax 61 2 938 27318




From: Robert Champaign :      r-champaign-at-raytheon.com
Date: Mon, 21 Feb 2000 08:05:00 -0800
Subject: Cool Prep (Water Coolers)

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


We are currently using Cool Prep in our Coolwell cooler system. This is a
liquid cleaner & slime preventer that helps keep mineral deposits to a
minimum in the cooling system. It also helps prevent corrosion in the
water passages without harming hoses, gaskets or seals. Since Coolwell is
no longer in business does anyone know of a replacement product for Cool
Prep.

Can we simply switch to ethylene glycol?
Robert Champaign
Raytheon
Electronic Systems Labs - Texas Region
r-champaign-at-raytheon.com
2501 W.University, MS 8011
McKinney Texas, 75070
972-952-3165




From: Michael Coviello :      coviello-at-mae.uta.edu
Date: Mon, 21 Feb 2000 09:58:54 -0600
Subject: SEM-JEOL 845 TMP problems

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hi Y'all:
We have an JEOL 845 SEM (TMP pumped) and we have been having problems with the
TMP REV light staying on. Because of the microscopes' timing circuit, the
scope will shut off after 20 minutes. The shutting off/timing issue can be
overridden by keeping the key turned clockwise: the microscope will
eventually go to the correct vacuum, but the darn TMP light never goes out.
JEOL told us of a trick of depressing the standby button on the TMP
controller, sometimes remedies this situation, but it didn't work in our
case. We have been in contact with JEOL and so far we/they haven't figured
out the problem. The TMP pump and the rough pump both seem OK: the TMP 80 %
relay kicks in at 80% RPM as it's supposed to and we changed the rough pump
oil, just to be sure). I have a few questions to ask of you 845 owners:
1) Have you seen this type of problem, and if so, how did you remedy it?
2) Does anyone out there have wiring diagrams, manuals for the TMP 845 (not
the DP, we already have that), if so can we get a copy? JEOL doesn't seem to
have anything.

Your help will be greatly appreciated.
Mike Coviello
Lab Manager
UT Arlington
Arlington, TX
817 272-5496





From: Ziel, R. (Rainer) :      Rainer.Ziel-at-akzonobel.com
Date: Mon, 21 Feb 2000 19:55:17 +0100
Subject: LM: Video-Adapter for Zeiss Photo Microscope

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


We have an old Photomicroscope from Zeiss. It's grey in color and about 20
years old, but works pretty good. Until now we have an old 1 inch video
camera on top. We want to change this video camera to a state of art
CCD-camera with 1/2 inch CCD-chip. To enlarge the field of view, we want to
use a video adapter with a magnification 0.5x. Unfortunately several
oprtical corrections are made within this adapter. Such an adapter is not
offered by Zeiss. Does anybody out there knows a company that sells such a
special adapter or can build one?

Kind Regards

Rainer

--------

Rainer Ziel
Acordis Analytical Services Obernburg
63784 Obernburg
Germany
Phone: 06022/812645
email: rainer.ziel-at-acordis.com






From: Ziel, R. (Rainer) :      Rainer.Ziel-at-akzonobel.com
Date: Mon, 21 Feb 2000 19:55:17 +0100
Subject: LM: Video-Adapter for Zeiss Photo Microscope

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


We have an old Photomicroscope from Zeiss. It's grey in color and about 20
years old, but works pretty good. Until now we have an old 1 inch video
camera on top. We want to change this video camera to a state of art
CCD-camera with 1/2 inch CCD-chip. To enlarge the field of view, we want to
use a video adapter with a magnification 0.5x. Unfortunately several
oprtical corrections are made within this adapter. Such an adapter is not
offered by Zeiss. Does anybody out there knows a company that sells such a
special adapter or can build one?

Kind Regards

Rainer

--------

Rainer Ziel
Acordis Analytical Services Obernburg
63784 Obernburg
Germany
Phone: 06022/812645
email: rainer.ziel-at-acordis.com






From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 15:10:21 -0600
Subject: Administrivia: Server is acting strange...

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Colleagues...

Yes I know the server appears to be acting up (i.e. not sending
any mail out).. I'm looking into the problem.

Nestor
Your Friendly Neighborhood SysOp






From: bozzola-at-siu.edu (John J. Bozzola)
Date: Mon, 21 Feb 2000 16:59:58 -0600
Subject: LM: COURSES

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


FORWARDED TO LISTSERVER,
PLEASE RESPOND TO THE ADDRESS SHOWN BELOW:

{randy_anderson-at-ameritech.net}

I am a member of MSA and I work for the University of Chicago in the
section of Nephrology. I was wondering if you would know of any
universities or other types of institutions that would offer summer
courses for the beginner and advance areas of immunofluorescence
microscopy. Any help that you can give me in matter would greatly be
appreciated.

Thank you

Randy Anderson

####################################################################
John J. Bozzola, Ph.D., Director
Micro-Imaging and Analysis Center
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
####################################################################






From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 17:07:02 -0600
Subject: Testing using TestList

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


This is a test to 3 zaluzec addresses and the archive.
5:06 pm






From: Earl Weltmer :      earlw-at-pacbell.net
Date: Mon, 21 Feb 2000 15:17:47 -0800
Subject: Re: SEM-JEOL 845 TMP problems

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


I have a copy can send you.

The JEOL Schematics are somewhat ambiguous as the logic from the turbo pump
activates several relays. The relays are NOT connected in the manner that one
would think, i.e. when the turbo pump speed is 80% the relay is activated and
signals the vacuum logic that all is well.

Earl Weltmer

Michael Coviello 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 Y'all:
} We have an JEOL 845 SEM (TMP pumped) and we have been having problems with the
} TMP REV light staying on. Because of the microscopes' timing circuit, the
} scope will shut off after 20 minutes. The shutting off/timing issue can be
} overridden by keeping the key turned clockwise: the microscope will
} eventually go to the correct vacuum, but the darn TMP light never goes out.
} JEOL told us of a trick of depressing the standby button on the TMP
} controller, sometimes remedies this situation, but it didn't work in our
} case. We have been in contact with JEOL and so far we/they haven't figured
} out the problem. The TMP pump and the rough pump both seem OK: the TMP 80 %
} relay kicks in at 80% RPM as it's supposed to and we changed the rough pump
} oil, just to be sure). I have a few questions to ask of you 845 owners:
} 1) Have you seen this type of problem, and if so, how did you remedy it?
} 2) Does anyone out there have wiring diagrams, manuals for the TMP 845 (not
} the DP, we already have that), if so can we get a copy? JEOL doesn't seem to
} have anything.
}
} Your help will be greatly appreciated.
} Mike Coviello
} Lab Manager
} UT Arlington
} Arlington, TX
} 817 272-5496





From: Christoph Bauer :      cbauer-at-midway.uchicago.edu
Date: Mon, 21 Feb 2000 17:33:42 -0800
Subject: Position available: EM-TECHNICIAN

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


ELECTRON MICROSCOPY TECHNICIAN
A full time position for a senior research technician is available at
the University of Chicago in the Department of Molecular Genetics and
Cell Biology. The lab of Elaine Fuchs Ph.D. is interested in the
cytoskeleton, in cell-cell adhesion and in the cell biology of skin
development.
The successful candidate will perform routine transmission electron
microcopy, including specimen acquisition, tissue processing,
ultramicrotomy, operation of a Philips CM120 electron microscope and
darkroom procedures/digital imaging. In addition he/she will be trained
in on-section immunolabeling using Lowicryl K4M.
Applicant Qualifications: Minimum requirements are a Bachelors degree
and 2 years of relevant experience.
If you are motivated to work in an excellent scientific environment with
state of the art instrumentation please contact:

Christoph Bauer Ph.D.
University of Chicago,
Department of Molecular Genetics and Cell Biology
5841 S. Maryland Ave/MC 1028
Chicago, Il 60637

Phone: 773 834 2896
Fax: 773 702 0141
Email:cbauer-at-midway.uchicago.edu




From: Robert Dourmashkin :      robert-at-bobsydnee.demon.co.uk
Date: Mon, 21 Feb 2000 18:18:47 -0600
Subject: MT2 Microtome

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html



Does anybody need a MT2 Porter-Blum ultramicrotome, in running order but
no binocular? Yours for the asking and transport.

Robert Dourmashkin,
Section of Academic Virology,
Royal London School of Medicine,
40 Turner Street,
Whitechapel London E1 2AD.
--
Robert Dourmashkin






From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 18:16:54 -0600
Subject: Administrivia: Restarting the Server

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Colleagues...

I'm reinitializing the server. You may see some old and/or
duplicate messages. I will try to restore those that I think
did not get through. If you tried to post a message in the last
day or so and haven't seen it please wait ~ 12 hours after
that time if you do not see it appearby then , you should
presume it was lost and repost your original
question/comment.

Sorry...

Nestor
Your Friendly Neighborhood SysOp






From: Greg Erdos :      gwe-at-biotech.ufl.edu
Date: Mon, 21 Feb 2000 18:18:12 -0600
Subject: Fwd: SEM/TEM Course

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html




} } Ms. Lalini Pillayi Would like to take both a TEM and SEM short
} } course. She is located at the at the US ARmy Dental Research Great
} } Lakes Lab.
} } Her applications will be both biological and biological
} } combined with material. If you offer such a course please contact her
} } directly at
} } lalini70-at-hotmail.com
} }
} } Thanks

Gregory W. Erdos, Ph.D. Ph. 352-392-1295
Assistant Director, Biotechnology Program
PO Box 110580 Fax:
352-846-0251
University of Florida
Gainesville, FL 32611








From: Earl Weltmer :      earlw-at-pacbell.net
Date: Mon, 21 Feb 2000 18:17:32 -0600
Subject: Re: SEM-JEOL 845 TMP problems

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html




I have a copy can send you.

The JEOL Schematics are somewhat ambiguous as the logic from the turbo pump
activates several relays. The relays are NOT connected in the manner that one
would think, i.e. when the turbo pump speed is 80% the relay is activated and
signals the vacuum logic that all is well.

Earl Weltmer

Michael Coviello 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 Y'all:
} We have an JEOL 845 SEM (TMP pumped) and we have been having problems
} with the
} TMP REV light staying on. Because of the microscopes' timing circuit, the
} scope will shut off after 20 minutes. The shutting off/timing issue can be
} overridden by keeping the key turned clockwise: the microscope will
} eventually go to the correct vacuum, but the darn TMP light never goes out.
} JEOL told us of a trick of depressing the standby button on the TMP
} controller, sometimes remedies this situation, but it didn't work in our
} case. We have been in contact with JEOL and so far we/they haven't figured
} out the problem. The TMP pump and the rough pump both seem OK: the TMP 80 %
} relay kicks in at 80% RPM as it's supposed to and we changed the rough pump
} oil, just to be sure). I have a few questions to ask of you 845 owners:
} 1) Have you seen this type of problem, and if so, how did you remedy it?
} 2) Does anyone out there have wiring diagrams, manuals for the TMP 845 (not
} the DP, we already have that), if so can we get a copy? JEOL doesn't seem to
} have anything.
}
} Your help will be greatly appreciated.
} Mike Coviello
} Lab Manager
} UT Arlington
} Arlington, TX
} 817 272-5496






From: Fred Pearson :      eoptics-at-mcmail.cis.mcmaster.ca
Date: Mon, 21 Feb 2000 18:18:24 -0600
Subject: Kevex Preamp Manual

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html



Good morning:

We would like to resurrect 2 older Kevex SiLi detectors for scintillation
counting, and have lost or misplaced the manuals for the preamp output
connectors, ie. voltages and pinouts.
The model numbers of the preamps are 2002 and 2003.

If anyone has this info, could you please fax or email?

Thanks in advance

Fred Pearson

********************************************************
Fred Pearson
Brockhouse Institute for Materials Research
McMaster University
1280 Main St. West
Hamilton, Ontario
Canada L8S 4M1


email: eoptics-at-mcmaster.ca
phone: (905) 525-9140 ext. 24609
fax: (905) 521-2773
********************************************************






From: Christoph Bauer :      cbauer-at-midway.uchicago.edu
Date: Mon, 21 Feb 2000 18:21:28 -0600
Subject: Position available: EM-TECHNICIAN

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html



ELECTRON MICROSCOPY TECHNICIAN
A full time position for a senior research technician is available at
the University of Chicago in the Department of Molecular Genetics and
Cell Biology. The lab of Elaine Fuchs Ph.D. is interested in the
cytoskeleton, in cell-cell adhesion and in the cell biology of skin
development.
The successful candidate will perform routine transmission electron
microcopy, including specimen acquisition, tissue processing,
ultramicrotomy, operation of a Philips CM120 electron microscope and
darkroom procedures/digital imaging. In addition he/she will be trained
in on-section immunolabeling using Lowicryl K4M.
Applicant Qualifications: Minimum requirements are a Bachelors degree
and 2 years of relevant experience.
If you are motivated to work in an excellent scientific environment with
state of the art instrumentation please contact:

Christoph Bauer Ph.D.
University of Chicago,
Department of Molecular Genetics and Cell Biology
5841 S. Maryland Ave/MC 1028
Chicago, Il 60637

Phone: 773 834 2896
Fax: 773 702 0141
Email:cbauer-at-midway.uchicago.edu






From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 18:22:34 -0600
Subject: JEOL 845 SEM (TMP Light)

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html



Hi Y'all:
We have an JEOL 845 SEM (TMP pumped) and we have been having problems with the
TMP REV light staying on. Because of the microscopes' timing circuit, the
scope will shut off after 20 minutes. The shutting off/timing issue can be
overridden by keeping the key turned clockwise: the microscope will
eventually go to the correct vacuum, but the darn TMP light never goes out.
JEOL told us of a trick of depressing the standby button on the TMP
controller, sometimes remedies this situation, but it didn't work in our
case. We have been in contact with JEOL and so far we/they haven't figured
out the problem. The TMP pump and the rough pump both seem OK: the TMP 80 %
relay kicks in at 80% RPM as it's supposed to and we changed the rough pump
oil, just to be sure). I have a few questions to ask of you 845 owners:
1) Have you seen this type of problem, and if so, how did you remedy it?
2) Does anyone out there have wiring diagrams, manuals for the TMP 845 (not
the DP, we already have that), if so can we get a copy? JEOL doesn't seem to
have anything.

Your help will be greatly appreciated.
Mike Coviello
Lab Manager
UT Arlington
Arlington, TX
817 272-5496






From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 18:23:47 -0600
Subject: Prep of rat brains for TEM using vascular perfusion

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Heather A Owen wrote:
}
} Hello everyone,
}
} A graduate student is getting ready for a rather large scale preparation
} of rat brains for TEM using vascular perfusion. Luckily (since I'm a
} botanist) her major professor has had experience with the procedure. He
} doesn't know for sure whether he used EM or Biological grade
} glutaraldehyde previously, but considering the large volume that will be
} required, and the cost difference, I'm hoping that the Biological grade
} will suffice.
}
} I would appreciate advice from those of you that have experience with this
} method of fixation.
}
} Thank you in advance for any information.
}
} Heather Owen
}
} Heather A. Owen, Director
} Electron Microscope Laboratory
} Department of Biological Sciences
} University of Wisconsin - Milwaukee
} (414)229-6816

Heather,

It's been a while but, back when I did such things, I had bouts of
non-uniform fixation when using biological grade glutaraldehyde solutions
even in plant tissues. However, the solutions had been kept much longer than
I'd now use and there are other preparation technique changes that confound
the determination of causality. In the cases where I've perfused small
animals, I've used EM grade GA since, as expensive as it is, it's cheaper
than repeating the experiment.

However, I did a full systemic perfusion. One random thought ( I haven't
tried it): What's the vascular volume of a rat brain? If you just need just
the brain, I'd think a little creative clamping (really small hemostats?) in
the thorax should let you reduce the volume needed to make even a large
experiment affordable with "the good stuff".

cheers and good luck,
John

John Heckman
MSM Department
Michigan State University







From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 18:24:19 -0600
Subject: OIL for use in JEOL's

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Santovac is a great oil to use in Jeol instruments. Backstreaming is
less and it will withstand a lot more abuse than Lion oil if the vac
system has a glitch. Just put a little less charge in than the normal Lion
oil. Regards Steve Parkins








From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 18:25:09 -0600
Subject: More on Rat Brain Prep..

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We use Biological grade to perfuse rats (whole body, we haven't tried clamping
bits off, though that might be a good idea since we're interested in just the
head). After the perfusion is finished, we remove the interesting bit, which is
huge by EM standards, and fix some more in Karnowski's. Then we go on to
osmium,
and continue processing. We get acceptable EM pictures at high magnification,
even though our blocks are so large and processing times are therefore
extended. So I can recommend Biological grade glutaraldehyde. Hope this helps.

Lesley Weston.







From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 18:25:57 -0600
Subject: CCD Camera on Photomicroscope from Zeiss

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We have an old Photomicroscope from Zeiss. It's grey in color and about 20
years old, but works pretty good. Until now we have an old 1 inch video
camera on top. We want to change this video camera to a state of art
CCD-camera with 1/2 inch CCD-chip. To enlarge the field of view, we want to
use a video adapter with a magnification 0.5x. Unfortunately several
oprtical corrections are made within this adapter. Such an adapter is not
offered by Zeiss. Does anybody out there knows a company that sells such a
special adapter or can build one?

Kind Regards

Rainer

--------

Rainer Ziel
Acordis Analytical Services Obernburg
63784 Obernburg
Germany
Phone: 06022/812645
email: rainer.ziel-at-acordis.com







From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 18:26:40 -0600
Subject: geometry settings for a Kevex Quantum detector for JEOL 2000FX

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Does anyone have the geometry settings for a Kevex Quantum detector on a
JEOL 2000FX? I am using DTSA and need the values. I would like to have the
sample to detector distance and the azimuthal angle. I am using +45 for the
azimuthal angle and 90 for the detector angle. Are these correct? I know
that the takeoff angle is 70.
-Scott


Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)







From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 18:27:29 -0600
Subject: looking for used ion mill

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Hello microscopy listserver-type people,

I am in the market for a used ion mill. Can anybody point me to one?

Thanks,
Larry

--
------------------------------------------------------------------------------
Larry R. Nittler Laboratory for Extraterrestrial Physics,
Code 691
Interstellar Dust Buster NASA Goddard Spaceflight Center
Greenbelt MD 20771
phone: 301-286-4572 fax:301-286-0212
nittler-at-lepvax.gsfc.nasa.gov http://www.ciw.edu/lrn/
------------------------------------------------------------------------------






From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 18:28:06 -0600
Subject: RE: autofluorescence quenching:spectral imaging can help

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} From: RLevenson-at-cri-inc.com Subject: RE: autofluorescence quenching:
spectral imaging can help

You asked about strategies for reducing autofluorescence.

In addition to various strategies which can include using near-IR dyes like
Cy5.5 and exciting far in the red, you can also treat tissues with sodium
borohydride or toluidine blue (these latter observations I pass on from
others--have not tried them myself).

I have recently had very promising results using spectral imaging to
discriminate autofluorescence from desired fluorescence. [Caution: I am
describing a product sold by my company]. CRI makes a liquid crystal
tunable filter (LCTF) that can be used to collect a stack of images at
different wavelengths. This yields a spectrum at every pixel of an image.
(A product with similar capabilities is sold by Applied Spectral Imaging,
but uses a different technology.) CRI's new LCTF, the FluoroSpec, uses
special optics to combine polarization states; along with other
improvements, it has more than twice the light throughput compared to
previous models.

The sample was formalin-fixed brain probed with a Cy2-labeled antibody to a
low-abundance protein. By eye it was very difficult to distinguish the Cy2
signal from the bright green to green-orange autofluorescence. Using a
spectral data set (15 images from 510 nm to 650 nm at 10 nm intervals) and
linear combination algorithms for pixel-unmixing, it was possible to
completely separate the Cy2 and autofluorescence images.

I would be glad to share these images with any interested parties.

Richard


Richard Levenson, MD
Technical Director, Biomedical Systems
Cambridge Research & Instrumentation (CRI), Inc.
80 Ashford St.
Boston, MA 02134

Ph: (617) 787-5700; Fax: (617) 787-4488
rlevenson-at-cri-inc.com www.cri-inc.com






From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 18:29:00 -0600
Subject: Help on fluorescence

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Hi all. I have just joined this list and am posting without the usual
"lurking time" due to time constraints - please forgive if recently
covered. I am also a novice in all microscopy techniques, especially
fluorescence.

I am trying to detect GFP-containing cells in liver tissue and having
some problems.

1. I can't determine the spectra for the filter blocks I am using (on a
Nikon Eclipse TE300 with TE-FM Epifluorescence attachment). The blocks
are marked "UV", "B-2" (blue from source, green in field) and "G" (green
from source, red in field). I have e-mailed Nikon and to be fair
they've only had a few days but I'm under some pressure. No help from
the manual. I have been using B-2 for GFP.

2. I get quite a lot of background fluorescence even with frozen
sections (fixed in neutral buffered formalin). Can anyone suggest a way
to reduce this? (eg any extra filters?)

3. I have read conflicting opinions on fixation. Most previous work has
used thick sections (50 microns cut eg with Vibratome, ? to avoid having
to embed tissue blocks). GFP is interfered with by acetone (and probably
other organic solvents) but I would have thought that after fixation
with formalin it should be stabilized and resistant to the
xylene/alcohol used in paraffin embedding. I have seen fluorescence
retained after this treatment but perhaps it can be improved.

4. For those with liver fluoro experience - on "UV" setting I see bright
fluorescence which photobleaches. I believe I am looking at retinoids in
stellate cells, although the bleaching is incomplete and a bit slower
than I have seen before. Can anyone confirm this?

Apologies for long post. Any help much appreciated.

David Lockwood
University of Qld Dept of Surgery
PA Hospital Brisbane Australia







From: A. Greene :      ablue-at-io.com
Date: Monday, February 21, 2000 10:02 AM
Subject: Cool Prep (Water Coolers)

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Hello Robert,
Preparation of the water in water chillers is a strange world of its own and
you may get many responses to your question. The majority of my experience
has been with the Haskris systems but the chemistry may be the same. In
about 30 years of looking after many chillers, my formula for success goes
like this:
1 - Use deionized or distilled water. The water will leach enough products
from the hoses, microscope lenses, pumps and so forth to reach a equilibrium
and then be "ideally suited" for your system.
2 - Don't use any fungicide or other bug killer because it will slime up the
works.
3 - Put 4 or 5 drops of oil on top of the water reservoir. This will form a
barrier by creating a monolayer on top of the water and keep things from
growing in the water.
I have used this proceedure in tropical climates and in the north and in
both dry and humid environments and it has always worked very well. I can
almost gurantee you that ethyline glycol will gum up the works.

Best wishes and good luck.

Alex Greene
SCIENTIFIC INSTRUMENTATION SERVICES, INC.
PMB-499, 1807 West Slaughter Lane, Number 200
Austin, Texas 78748-6200 Phone:512-282-5507 FAX 512/280-0702

QUALITY ELECTRON MICROSCOPE REPAIRS


-----Original Message-----
} From: Robert Champaign {r-champaign-at-raytheon.com}
To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}





From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Mon, 21 Feb 2000 23:38:16 -0600
Subject: looking for a bad address

Contents Retrieved from Microscopy Listserver Archives
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This is a short test

11:38 pm






From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Tue, 22 Feb 2000 00:21:26 -0600
Subject: another test but to microscopy@sparc5.microscopy.com

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another test but to microscopy-at-sparc5.microscopy.com






From: Jo Verbeeck :      joverbee-at-ruca.ua.ac.be
Date: Tue, 22 Feb 2000 10:52:37 +0100 (MET)
Subject: TEM EELS:How to focus ESI images?

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Hello all,

I'm using a CM30 FEG with GIF PEELS atached to it. I'm getting confused
about focussing an energy filtered image.

GATAN proposes to focus the image on the TV camere at a loss of
100eV.Apparently it will be focused more or less OK at a higher loss say
741eV.
DM uses the voltage offset which changes the HT of the microscope. So as
far as I understand the focus should be OK for every energy loss(no
refocussing should be needed at all) . Although in practice it is not. The
trick with focussing at 100eV works better than not focussing at all but
it is not satisfactory for high spatial resolution.

In the Phd of M.Schenner (TU Wien "The quantification problem in EELS
Microanalysis") I found a callibration procedure to find a defocus to
correct for chromatic abberation as a function of the energy loss you are
interested in. But he seems to use the drift tube in the GIF.

The question now: How should I focus my ESI images and what is the
explanation behind it?

Thanks in advance,

Jo

*************************************************************
* Jo Verbeeck *
* University of Antwerp *
* Dept. EMAT (Electron Microscopy for Materials Research) *
* e-mail: joverbee-at-ruca.ua.ac.be *
* tel: +32(0)3 218 02 49 *
* fax: +32(0)3 218 02 57 *
*************************************************************





From: ALEX BLACK :      ALEXANDER.BLACK-at-NUIGALWAY.IE
Date: Tue, 22 Feb 2000 11:29:17 +0000 (GMT)
Subject: Quinacrine fluorescence

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Dear All,
We are considering trying to label aspects of innervation using a
quinacrine fluorescence technique. So far the preliminary trials have not gone
well. Could anyone give advice on this, especially on a source for a reliable
fluorescent quinacrine dye?


Thanks in advance

Alex Black
Department of Anatomy
National University of Ireland, Galway





From: Barbara Foster :      mme-at-map.com
Date: Tue, 22 Feb 2000 08:11:57 -0500
Subject: Re: LM: COURSES

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Dear Randy,

We have specialists in this area who can offer customized, on-site courses,
using your equipment and speaking specifically to your application. If you
are interested in further details, or in how U. Chi can sponsor such a
course, please contact me directly.

Best regards,
Barbara Foster
Consortium President
Microscopy/Microscopy Education ...Educating microscopists for greater
productivity.

125 Paridon Street Suite 102 Springfield, MA 01118
PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Visit our web site {http://www.MME-Microscopy.com/education}


At 04:59 PM 2/21/00 -0600, John J. Bozzola wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America




From: Debby Sherman :      sherman-at-btny.purdue.edu
Date: Monday, February 21, 2000
Subject: Re: Prep of rat brains for TEM using vascular perfusion

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I perfused dozens of rat brains over the years. I would recomend using the purified glutaraldehyde. However, you can use a slightly less concentrated solution since the fixation is so much more efficient tham immersion fixation. We primarily fixed for ICC but only the choice of fix would vary not the method. For normal ultrastructure, I would try 2-3% Glut or half-strength Karnovsky's in a phosphate buffer system with added NaCl and Mg at physiological concentrations and see which fixation I prefered by examining the desired tissues prior to fixing critical animals.

We would anesthetize the animal, open the chest cavity and clamp off the descending aorta, thus limiting the perfusion to the upper half of the animal. The cannula would then be inserted into a small slit in the left ventricle, positioned up into the aorta, and held with a hemostat. Perfusion lines should have been filled with normal saline or ringers so that as soon as the cannula is in place, the flow can be started to wash out the blood. A small slit is made in the right atrium to permit release of venous blood, etc. As soon as the return is relatively pale, switch to your fixative and run the desired amount through. The condition of the liver is a reasonable criteria for the efficiency of the perfusion. It should be very pale yellow as the blood is removed and should stiffen up within a few minutes. In fact the entire upper part of the animal should become quite stiff as the tissues are fixed. Figure on a couple of hundred mls. of fix per animal.

A couple of suggestions are to use the initial ringer wash and fixatives at room temperature or slightly above so as not to cause vessels to contract. Do any further washes and post fix the portions of interest at 4oC. Also make sure you have a consistant but gental perfusion rate so as not to break delicate capillaries. Usually this can be accomplished just by raising the vessels containing the buffer and fix above the animal and let gravity do the rest.
Good luck,
Debby

Debby Sherman, Manager Phone: 765-494-6666
Microscopy Center in Agriculture FAX: 765-494-5896
Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu
Purdue University
1057 Whistler Building
West Lafayette, IN 47907-1057

--------------------------------------



Heather A Owen wrote:
}
} Hello everyone,
}
} A graduate student is getting ready for a rather large scale preparation
} of rat brains for TEM using vascular perfusion. Luckily (since I'm a
} botanist) her major professor has had experience with the procedure. He
} doesn't know for sure whether he used EM or Biological grade
} glutaraldehyde previously, but considering the large volume that will be
} required, and the cost difference, I'm hoping that the Biological grade
} will suffice.
}
} I would appreciate advice from those of you that have experience with this
} method of fixation.
}
} Thank you in advance for any information.
}
} Heather Owen
}
} Heather A. Owen, Director
} Electron Microscope Laboratory
} Department of Biological Sciences
} University of Wisconsin - Milwaukee
} (414)229-6816

Heather,

It's been a while but, back when I did such things, I had bouts of
non-uniform fixation when using biological grade glutaraldehyde solutions
even in plant tissues. However, the solutions had been kept much longer than
I'd now use and there are other preparation technique changes that confound
the determination of causality. In the cases where I've perfused small
animals, I've used EM grade GA since, as expensive as it is, it's cheaper
than repeating the experiment.

However, I did a full systemic perfusion. One random thought ( I haven't
tried it): What's the vascular volume of a rat brain? If you just need just
the brain, I'd think a little creative clamping (really small hemostats?) in
the thorax should let you reduce the volume needed to make even a large
experiment affordable with "the good stuff".

cheers and good luck,
John

John Heckman
MSM Department
Michigan State University










From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Tue, 22 Feb 2000 07:38:34 -0600
Subject: Administrivia: Still debugging

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Folks...

Still in a debug mode this is a very peculiar problem.
This test is going to the entire distribution..

Sorry

Nestor






From: John Catino :      jwcatino-at-concentric.net
Date: Tue, 22 Feb 2000 19:42:06 -0500
Subject: Enlarger and Print Processor Sale

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I have a Beseler 45M Enlarger and Kodak Royal Print Processor for sale.
Both are in working condition. We are going digital! Equipment is
located in Easton, PA.




John Catino






From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Tue, 22 Feb 2000 18:42:08 -0600
Subject: another test of Microscopy

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This is a second test without the 15 minute delay/queus






From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Tue, 22 Feb 2000 18:47:07 -0600
Subject: Testing at 6:47

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Testing at 6:47 to Microscopy






From: Nestor J. Zaluzec :      zaluzec-at-sparc5.microscopy.com
Date: Tue, 22 Feb 2000 19:20:37 -0600
Subject: Administrivia: DNS restored (I think)

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Colleagues....

Well the DNS (Domain Name Server) failed and was probably
pretty sick over most of last week. Alot of mail probably just
ended up in a black hole somewhere on the Net. Some of you
may have gotten messages, but I see that most of the mail
I sent out during my trip DownUnder to the ACEM-16 meeting
in Canberra never made it to it's destination.

The DNS "looks" fixed now but I'm not confident enough to proclaim
all as back to normal yet. Let's see if this message actually
gets through to everyone this time.

Sorry gang....

Nestor
Your Friendly (Frustrated) Neighborhood SysOp






From: Toby Knight :      tknight-at-waite.adelaide.edu.au
Date: Wed, 23 Feb 2000 13:32:59 +1030 (CST)
Subject: Sudan Black staining and fixing

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Greetings,

I have 2 queries pertaining to the use of the lipid colourant Sudan Black
B, I hope someone can be of some help:

1. I have been able to stain oil traces in the glands of citrus fruit BUT
the epoxy resin Procure/Araldite also stains quite strongly. Is this
typical of epoxy resins and would Spurrs show the same problem?

- Should I reduce the staining time from 30 mins?
- Adjust the temperature from 60 degrees C for staining?
- Rinse my sections for longer with 70% ethanol after staining?

2. I have used osmium tetroxide to fix lipids in the citrus rind tissues,
with mixed results.
I have also seen some success in oil fixation by applying Sudan Black B
to the tissue prior to chemical fixation. I assume it binds and reduces
the oil solubility. I am going to test this by treating tissue with SBB
prior and post chemical fix, and comparing.

- SBB is made up in 70% ethanol. As a postfix, would it be most logical
to introduce it at the 70% step in my ethanol dehydration series? (10 to
100%)

thanks kindly, Toby Knight (struggling PhD).


--------------------------------------------------------
Toby Knight
PhD student

Department of Horticulture, Viticulture and Oenology
The University of Adelaide
Plant Research Centre
Waite Campus, PMB 1
Glen Osmond, SA 5064
AUSTRALIA

Tel: +61 8 8303 6668 or 8303 6665
HVO: +61 8 8303 7242
Fax: +61 8 8303 7116
Email: tknight-at-waite.adelaide.edu.au
--------------------------------------------------------






From: Dr. Manfred Rohde :      mro-at-GBF.de
Date: Wed, 23 Feb 2000 07:26:57 +0100
Subject: Immuno-labeling of glutaraldehyd, osmium embedded samples in

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Dear Microscopist,

has anyone out there any experiences with immuno-labeling of osmificated and
Spurr embedded samples. We have tried out etching, oxidizing sections with
hydrogen peroxide (10% for 15 min) or sodium methylat (1% for 15 min)
without any significant increase of label intensity (Ab used is polyclonal
from rabbit, protein A purified)

Any suggestions are welcome. Manfred





From: Fatima Merchant :      merchant-at-persci.com
Date: Wed, 23 Feb 2000 07:48:45 -0600
Subject: Re: pinhole grids

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Nestor,

It made it her ok. I thought I was getting pretty good
service all week. But I know what evils hide in name
servers. Not nearly to the extent you do:)

As always you efferots are welcome. You provide the
pipe for one of the best resorces on the net.

Gordon

G. C,. Couger
624 Cheyenne
Stillwater, OK 74075-1411
405 624 2855 between 3:00 pm to 1:00 am CST
405 742 2758 cell
----- Original Message -----
} From: "Nestor J. Zaluzec" {zaluzec-at-sparc5.microscopy.com}
To: {Microscopy-at-sparc5.microscopy.com}
Sent: Tuesday, February 22, 2000 7:20 PM



}
} Dear All:
}
} I was wondering if anyone out there is aware of a company
} that sells pinhole grids. I am looking for grid(2D) of
} pinholes that are approximately 15-20 microns in diameter
} with a center to center separation of about 150-200 microns.
}
} I would greatly appreciate any information that you could
} provide regarding the commercial availability of similar
} pinhole grids.
}
} TIA,
} Fatima Merchant
}
}
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}
}
} Fatima Merchant, Ph.D.
} Senior Research Engineer
} Perceptive Scientific Instruments, Inc.
} 2525 South Shore Blvd., Suite 100
} League City, Texas 77573
}
} Telephone: (281) 334-3027 Ext: 230
} Toll Free: (800) 288-3027 Ext: 230
} Facsimile: (281) 538-2222
} Email: merchant-at-persci.com
}
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}
}
}






From: G Peranzi :      peranzi-at-bichat.inserm.fr
Date: Wed, 23 Feb 2000 08:25:18 -0600
Subject: KNEWS ABOUT GLOW DISCHARGE ACCESSORY

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-----Original Message-----
} From: Valdes Erica R Dr. SBCCOM
Sent: Tuesday, February 22, 2000 1:26 PM
To: 'microscopy-at-sparc5.Microscopy.Com'


We got a sputter-coater system (VG Microtech SC500) that we would like to
convert
into a glow discharge. Does anybody have done it already?
We got electronic schemes to do the High Voltage unit control, but we miss the
mecanical design to fit into our vacuum chamber (of the SC500). Does anybody
know
how to do?
We know that an acessory (for the Microtech) have been developped some years
ago
(GD350A) but it is not anymore available. What are the modification made to
use this
chamber?
Jean-Jacques LACAPERE
INSERM U 410
Facult de Mdecine X. Bichat
Paris FRANCE






From: Jo Verbeeck :      joverbee-at-ruca.ua.ac.be
Date: Wed, 23 Feb 2000 08:25:41 -0600
Subject: TEM EELS:How to focus ESI images?

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Hello all,

I'm using a CM30 FEG with GIF PEELS atached to it. I'm getting confused
about focussing an energy filtered image.

GATAN proposes to focus the image on the TV camere at a loss of
100eV.Apparently it will be focused more or less OK at a higher loss say
741eV.
DM uses the voltage offset which changes the HT of the microscope. So as
far as I understand the focus should be OK for every energy loss(no
refocussing should be needed at all) . Although in practice it is not. The
trick with focussing at 100eV works better than not focussing at all but
it is not satisfactory for high spatial resolution.

In the Phd of M.Schenner (TU Wien "The quantification problem in EELS
Microanalysis") I found a callibration procedure to find a defocus to
correct for chromatic abberation as a function of the energy loss you are
interested in. But he seems to use the drift tube in the GIF.

The question now: How should I focus my ESI images and what is the
explanation behind it?

Thanks in advance,

Jo

*************************************************************
* Jo Verbeeck *
* University of Antwerp *
* Dept. EMAT (Electron Microscopy for Materials Research) *
* e-mail: joverbee-at-ruca.ua.ac.be *
* tel: +32(0)3 218 02 49 *
* fax: +32(0)3 218 02 57 *
*************************************************************






From: Greg Erdos :      gwe-at-biotech.ufl.edu
Date: Wed, 23 Feb 2000 09:37:47 -0500
Subject: Re: Immuno-labeling of glutaraldehyd, osmium embedded samples

Contents Retrieved from Microscopy Listserver Archives
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Material fixed like yours works only rarely. You might try etching with
saturated sodium metaperiodate for one hour followed by water washes before
incubating with the primary.
That is assuming that your antigen is not periodate sensitive.

At 07:26 AM 02/23/2000 +0100, you wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

Gregory W. Erdos, Ph.D. Ph. 352-392-1295
Assistant Director, Biotechnology Program
PO Box 110580 Fax:
352-846-0251
University of Florida
Gainesville, FL 32611






From: William Tivol :      tivol-at-wadsworth.org
Date: Wed, 23 Feb 2000 10:03:08 -0500
Subject: Re: pinhole grids

Contents Retrieved from Microscopy Listserver Archives
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Fatima Merchant wrote:

} } I was wondering if anyone out there is aware of a company
} } that sells pinhole grids. I am looking for grid(2D) of
} } pinholes that are approximately 15-20 microns in diameter
} } with a center to center separation of about 150-200 microns.
} }
} } I would greatly appreciate any information that you could
} } provide regarding the commercial availability of similar
} } pinhole grids.
} }

Dear Fatima,
I asked about the same question and got a reply from
Janko Otto (otto-at-quantifoil.com). He wrote me:

"I read a discussion about making holey films . We had
solving the problem using semiconductor lithographic techniques.

We have been producing QUANTIFOIL, a perforated foil with pre-definable
hole size shape and arrangement. The variation among the hole sizes is
small: the standard deviation for their distribution is well below 0.1
m within a batch; the reproducibility for the average hole size between
batches is 0.2 m. The shapes of the holes in the foils produced so far
are circular and square. Until now, their arrangements have been
orthogonal.

Foils with any desired hole size in the micrometer range, and with any
shape and arrangement can be fabricated. Any of these three parameter
can also be varied in a defined manner within one foil.

Until now, we produce three different types of QUANTIFOIL:

Type Hole Repetition distancee
um um
R1.2/1.3 ~1.2 2.5 (circular holes)
R2/2 ~2 4 (circular holes)
S7/2 ~7 9 (square holes)

QUANTIFOIL is generally delivered as a carbon foil; it can also be
obtained strengthened with plastic film.

If you interested in these perforated foil, I can send you some samples."

He may be able to produce what you want. I have not yet tried
his grids out, but I intend to when I get my specimens ready. I am not
affiliated with Quantifoil, just a potential user. Good luck.
Yours,
Bill Tivol







From: William Tivol :      tivol-at-wadsworth.org
Date: Wed, 23 Feb 2000 10:38:42 -0500 (EST)
Subject: Re: Cool Prep (Water Coolers)

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*************************************************************
* Jo Verbeeck *
* University of Antwerp *
* Dept. EMAT (Electron Microscopy for Materials Research) *
* e-mail: joverbee-at-ruca.ua.ac.be *
* tel: +32(0)3 218 02 49 *
* fax: +32(0)3 218 02 57 *
*************************************************************

---------- Forwarded message ----------


} We are currently using Cool Prep in our Coolwell cooler system. This is a
} liquid cleaner & slime preventer that helps keep mineral deposits to a
} minimum in the cooling system. It also helps prevent corrosion in the
} water passages without harming hoses, gaskets or seals. Since Coolwell is
} no longer in business does anyone know of a replacement product for Cool
} Prep.
}
} Can we simply switch to ethylene glycol?

Dear Robert,
The suggestion of using distilled water and letting the system
come to equilibrium might work in a truly closed system composed of only
one material, but if you need to add water, or if you have several metals
in the system--as we do--this will be unsatisfactory in the long run.
We have found that Aqua-Treet 42 works very well in our Haskris system.
This product is made by Aqua Laboratories, Inc., PO Box 645, 8 Indus-
trial Way, Amesbury MA, (978) 388-3989. I have no relationship to this
company except as a satisfied user of their product.
Yours,
Bill Tivol




From: Walck. Scott D. :      walck-at-ppg.com
Date: Wed, 23 Feb 2000 11:42:55 -0500
Subject: DTSA and EMMFF 1.1?

Contents Retrieved from Microscopy Listserver Archives
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I am running two copies of DTSA, one is an older version and one is the
newer one that will run on a power Mac. I am also fairly new to DTSA. The
old version does not have the MSA format capability for reading and writing
files, but the newer one does. However, the new one will read two versions
of MSA format, 1.0 and 1.1 but will only save in 1.1. Now I'm confused. I
asked Nestor where I could get the latest MSA description of the MSA format
and he sent me what I already have, EMMFF.total from of the EMMPDL/MAS-Lib
files. I know that Nestor was on that committee. So what gives with MSA
version 1.1 in the latest version of DTSA? Why don't they have the 1.0
option? The keywords are not even the same.



-Confused in Pittsburgh.


Scott D. Walck, Ph.D.
PPG Industries, Inc.
Glass Technology Center
Guys Run Rd. (packages)
P. O. Box 11472 (letters)
Pittsburgh, PA 15238-0472

Walck-at-PPG.com

(412) 820-8651 (office)
(412) 820-8161 (fax)






From: wft03-at-health.state.ny.us
Date: Wed, 23 Feb 2000 12:13:50 -0500
Subject: Re: Cool Prep (Water Coolers)

Contents Retrieved from Microscopy Listserver Archives
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Why not use a liquid that has NO water in it. That solved my problem in
vibratory polishing of dissimilar metals in an alumina slurry. Ethylene
glycol or propylene glycol, (less toxic), are possibilities.


Dear Bernie,

That will work unless there is some microorganism which will

grow in ethylene glycol. One could also just buy commercial anti-freeze

and get some additional corrosion protection. I don't think that there

would be any problem with the circulation pump, since the viscosity

of glycol is similar to that of water. Keeping the fluid in the system for

some decades has different requirements from using it over a period

of hours, though.

Yours,

Bill Tivol






From: Carl Henderson :      chender-at-umich.edu
Date: Wed, 23 Feb 2000 12:22:05 -0500
Subject: Re: DTSA and EMMFF 1.1?

Contents Retrieved from Microscopy Listserver Archives
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Look for a "Write MSA 1.0" plug-in in the "extras Folder" of your
DTSA download and move it to the "Plug-ins" folder at the same level
as DTSA.



At 11:42 AM -0500 2/23/00, Walck. Scott D. wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

======================================
Carl Henderson
Electron Microbeam Analysis Laboratory
University of Michigan
2501 C.C. Little Bldg.
Ann Arbor, MI 48109-1063 USA
(734) 936-1550 FAX (734) 763-4690
======================================




From: Tobias Baskin :      BaskinT-at-missouri.edu
Date: Wed, 23 Feb 2000 11:39:28 -0600
Subject: (FE)SEM, sputter coating puzzle

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Greetings,
I wonder if anyone could explain the following observation
to me. I have been sputter coating with platinum and viewing the
sample with a FESEM (Hitatchi 4700, below lens type). In a certain
type of sample, the image appears darker than the background where
there is sample. Now these samples are perhaps a bit unusual. They
are very thin, starting off life as 1.75 um methacrylate sections of
a plant root adhered to a glass coverslip, followed by extraction in
hot acidic peroxide to remove all organic material except crystalline
cellulose. The cellulosic cell walls left behind on the coverslip are
nominally 100 nm thick and perhaps flattened further by the
processing. The cell walls are present in the sample as either cross
sections or as small regions laying in the plane of the coverslip.
These regions can be anywhere from the size of a cell (10 x 40 or so
microns) down to small portions thereof. All of this remaining cell
wall is darker than the surrounding background. I am putting on a
thin coat of Pt, somewhere around 2 nm, and imaging at 2 kV (I have
seen the same thing at 5 kV). It really puzzles me that my sample is
darker than background. Any explanations???

THanks,
Tobias
_ ____ ^ __ ____ Tobias I. Baskin
/ \ / / \ / \ \ University of Missouri
/ | / / \ \ \ Biological Sciences
/___/ /__ /___ \ \ \__ 109 Tucker Hall
/ / / \ \ \ Columbia, MO 65211-7400 USA
/ / / \ \ \ voice: 573-882-0173
/ /____ / \ \__/ \____ fax: 573-882-0123




From: Debby Sherman :      sherman-at-btny.purdue.edu
Date: 23 Feb 2000 13:17:19 -0500
Subject: Food Science Prep.

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------------------------------------------------------------------------
The Microscopy ListServer -- Sponsor: The Microscopy Society of America


Hi,
Well I guess we never run out of new challenges when it comes to specimen prep. This one concerns a product that is made of a cereal bound together with a sugar syrup. The investigators want to see how the syrup behaves as to structural integrety over a period of time with and without the addition of various stabilizing components. Information would be collected at the light microscopic level.
We first thought of cryosectioning followed by examination using LM. However, the product is filled with air pockets and literally disintegrates when microtomed. It is easily cut into pieces at temperatures above freezing as long as the pieces are no thinner than3-4 mmAccempts to infiltrate it with OCT compoound prior to freezing were not satisfactory.
New thought is to use a low temperature resin to infiltrate the sample followed by UV or chemical polymerization. butThe resulting blocks could then be microtomed at room temperature and imaged. It would be desirable to cut sections of 4-8 m in thickness which is too thick for an ultramicrotome with glass or diamond knives. Also UV polymerization would probably limit us to very small sample size which may be a problem.
Any suggestions would be appreciated as to resins to use which have low viscosity, could be used to infiltrate pieces of ~0.5-1cm sq, polymerize at low temperatures (+4 to 0oC range) using UV or chemical polymerization, and are preferably soft enough to section with a metal microtome blade. Any other suggestions of preparations techniques to try would be very appreciated.

Thanks in advance for your input.
Debby

Debby Sherman, Manager Phone: 765-494-6666
Microscopy Center in Agriculture FAX: 765-494-5896
Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu
Purdue University
1057 Whistler Building
West Lafayette, IN 47907-1057





From: Hank Adams :      hpadams-at-bcm.tmc.edu
Date: Wed, 23 Feb 2000 11:14:42 -0600
Subject: TEMs

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Dear listservers, I need to determine the value of a Philips 410 that has
been on service contract until it was decommissioned last year. I need this
information for an insurance claim. Is there a company or dealer of used em
equipment that can help me?
Thanks
Hank Adams
Integrated Microscopy Core
Molecular and Cellular Biology
Baylor College of Medicine
Houston, Tx





From: Timothy Schneider :      Timothy.Schneider-at-mail.tju.edu
Date: Wed, 23 Feb 2000 14:29:55 -0500
Subject: GLOW DISCHARGE

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Sorry if this is a second post, but we just got an e-mail saying the
first was rejected as spam, so here goes:






Hi:

I want to be able to "Glow Discharge" carbon coated copper grids for the
purpose of making them hydrophilic and negatively charged, so that I can
spread aqueous suspensions on them. I have a Denton Vacuum Desk ll sputter
coater with a "etch" mode. The question I have for my fellow list servers
is: will "etching" the grids do the same thing as "Glow Discharging" them?
Many Thanks, Tim


Timothy Schneider, Director of Electron Microscopy
Department of Pathology
Thomas Jefferson University
Room 229 Jeff Hall
1020 Locust St.
Philadelphia, Pa.
19107
215-503-4798
Timothy.Schneider-at-Mail.TJU.EDU





From: jillp-at-telecomedia.co.jp
Date: Wed, 23 Feb 2000 04:43:29 -0600
Subject: Version 5.0 is Now Available

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From: jillp-at-telecomedia.co.jp
Date: Wed, 23 Feb 2000 04:43:29 -0600
Subject: Version 5.0 is Now Available

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Now you can quickly and easily have access to the most powerful internet investigation sites and databases available today!

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The "N M S SOFTWARE" is absolutely astounding!

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Trace anyone by social security number!

Free Internet Access

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Unlisted phone numbers!

Get anyone's phone number with just a name - even unlisted numbers!

Locate!

Long lost friends, relatives, a past lover who broke your heart!

Send anonymous e-mail completely untraceable!

Investigate anyone!

Use the sources that private investigators use (all on the Internet)
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Ex-spouse!

Learn how to get information on an ex-spouse that will help you

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Criminal search-background check!

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Find out!

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From: Doug Cromey :      doug-cromey-at-ns.arizona.edu
Date: Wed, 23 Feb 2000 13:10:48 -0700
Subject: Can EELS, WDS or FT-IR localize As in Biol. tissues?

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Colleagues,

We are interested in localizing arsenic in biological tissues (toxicology
research), but have been unable to come up with a technique that will give
us the cellular localization information we're looking for (sub-cellular
would be even better).

We have tried TEM with energy dispersive x-ray microanalysis and our
concentrations are apparently too low, so we are unable to detect arsenic.

A few years ago we looked into using the autometallographic techniques that
have been published by Dr. Gorm Dansher & his colleagues. We were told
that the photographic emulsions required were no longer available.

In a brainstorming session yesterday several techniques were bantered
about, but in truth we are not familiar enough with the techniques to know
if they would suit our needs. Does EELS have anything to offer, or perhaps
wavelength dispersive spectoscopy (WDS)? How about some of the new FT-IR
instruments? Are there any other techniques that might be able to localize
arsenic and/or indicate its state of oxidation?

Vendors are welcome to reply.

Yours,
Doug Cromey
....................................................................
: 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: doug-cromey-at-ns.arizona.edu):
:...................................................................:
http://www.pharmacy.arizona.edu/exp_path.html
Home of: "Microscopy and Imaging Resources on the WWW"





From: Scott Wight :      scott.wight-at-nist.gov
Date: Wed, 23 Feb 2000 15:28:52 -0500
Subject: Re: DTSA and EMMFF 1.1?

Contents Retrieved from Microscopy Listserver Archives
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Scott & listers

Go to http://www.cstl.nist.gov/div837/837.02/dtsa.html and click on
"supporting files" page then download "preferences and plugins" you will
find 2 versions of the MSA write 1.0 (one is Kevex specific). It would
make life much easier on us microscopists if all manufacturers adopted the
MSA format.
Good luck,
Scott


} I am running two copies of DTSA, one is an older version and one is the
} newer one that will run on a power Mac. I am also fairly new to DTSA. The
} old version does not have the MSA format capability for reading and writing
} files, but the newer one does. However, the new one will read two versions
} of MSA format, 1.0 and 1.1 but will only save in 1.1. Now I'm confused. I
} asked Nestor where I could get the latest MSA description of the MSA format
} and he sent me what I already have, EMMFF.total from of the EMMPDL/MAS-Lib
} files. I know that Nestor was on that committee. So what gives with MSA
} version 1.1 in the latest version of DTSA? Why don't they have the 1.0
} option? The keywords are not even the same.
}
} -Confused in Pittsburgh.
}
} Scott D. Walck, Ph.D.
} PPG Industries, Inc.
} Glass Technology Center
} Guys Run Rd. (packages)
} P. O. Box 11472 (letters)
} Pittsburgh, PA 15238-0472
} Walck-at-PPG.com
} (412) 820-8651 (office)
} (412) 820-8161 (fax)

For the weather watchers: Mostly sunny, 60 F, snow is melted away.

-------------------note: new mailing address----------------------
Scott Wight e-mail: scott.wight-at-nist.gov
NIST 222/A113 W voice: 301-975-3949
100 Bureau Dr STOP 8371 | fax: 301-417-1321
Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed
is my own and does not represent those of my employer.






From: Supanee Danviriyakul :      supaneed-at-foodsci.umass.edu
Date: Wed, 23 Feb 2000 15:40:10 -0500 (EST)
Subject: Image analysis

Contents Retrieved from Microscopy Listserver Archives
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Dear all,
I am doing the image analysis of SEM micrographs. The
micrographs contain both dense (light) and air cell (dark) areas. But two
areas are not discrete. I'm having a difficulty of quantifying those
separated fractions. I wonder if anyone has any ideas of how to
solve the problem.
Looking forward for your suggestions.
Regards,
Supanee





From: Rejane Magalhes Pimentel
Date: Wed, 23 Feb 2000 19:25:00 -0300
Subject: ellipse at Image Tool

Contents Retrieved from Microscopy Listserver Archives
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Hello all,
I need to know how to draw an ellipse to measure at Image Tool software.
Thanks a lot.
Rejane Pimentel
Rejane Magalhes Pimentel Galindo
Functional Plant Morphology
Universidade Federal Rural de Pernambuco
ggalindo-at-elogica.com.br
Rua Maria Carolina, 417/804
51020-220 Boa Viagem





From: Sandra Perkins :      skperkin-at-vt.edu
Date: Wed, 23 Feb 2000 17:06:32 -0500
Subject: epoxy embedding of rat eye

Contents Retrieved from Microscopy Listserver Archives
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I thought I'd try this again in case it got lost in space:

Hi-

We are going to be processing rat eyes (from perfused animals) for
embedding in epoxy and I was wondering if anyone has any suggestions for
processing. We are interested in the retina, optic nerve, cornea and uveal
tract.

Thank you very much,
Sandy Perkins






From: Warren E Straszheim :      wesaia-at-iastate.edu
Date: Wed, 23 Feb 2000 16:03:17 -0600
Subject: Re: Image analysis

Contents Retrieved from Microscopy Listserver Archives
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Ahh! That is always the rub, trying to get the areas of interest to stand
out as a distinct range of gray scales.

If it is just a question of noise pixels in what is otherwise a distinct
region, then you might try filtering (smoothing) your image before
thresholding. Or you might try changing your image acquisition protocol to
get less noisy images.

You might try a different signal to find something that is less subject to
overlaps. Usually I resort to backscattered electron imaging since the
dependence on atomic number of that signal typically results in distinct
gray levels.

If that won't do it, you might be able to perform some sort of image
processing to separate the regions. There are techniques available, but
there seems to be as much art as science to them. I would refer you to John
Russ' "Image Processing Handbook" or a similar text, because there is not a
short answer to that question.

Warren Straszheim

At 03:40 PM 2/23/2000 -0500, you wrote:
} Dear all,
} I am doing the image analysis of SEM micrographs. The
} micrographs contain both dense (light) and air cell (dark) areas. But two
} areas are not discrete. I'm having a difficulty of quantifying those
} separated fractions. I wonder if anyone has any ideas of how to
} solve the problem.
} Looking forward for your suggestions.
} Regards,
} Supanee





From: Gordon Couger :      gcouger-at-rfdata.net
Date: Wed, 23 Feb 2000 18:37:38 -0600
Subject: Re: Cool Prep (Water Coolers)

Contents Retrieved from Microscopy Listserver Archives
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} From: {"wft03-at-health.state.ny.us"-at-sparc5.microscopy.com}
To} Why not use a liquid that has NO water in it. That solved my problem
in
} vibratory polishing of dissimilar metals in an alumina slurry. Ethylene
} glycol or propylene glycol, (less toxic), are possibilities.
}
}
} Dear Bernie,
}
} That will work unless there is some microorganism which will
}
} grow in ethylene glycol. One could also just buy commercial anti-freeze
}
} and get some additional corrosion protection. I don't think that there
}
} would be any problem with the circulation pump, since the viscosity
}
} of glycol is similar to that of water. Keeping the fluid in the system
for
}
} some decades has different requirements from using it over a period
}
} of hours, though.

Glycols won't have near the heat capacity of water. I doubt
you will find a bug that will live in ethylene glycol but you can
probably find many that can live in propylene glycol.

Any cooling system needs to have the coolant changed
periodically. Following the manufactures recommendation
is the safe way to go.

Gordon

Gordon Couger gcouger-at-couger.com

Stillwater, OK www.couger.com/gcouger
405 624-2855 GMT -6:00






From: Ladd Resaerch :      ladres-at-worldnet.att.net
Date: Wed, 23 Feb 2000 18:17:38 -0600
Subject: Re: pinhole grids

Contents Retrieved from Microscopy Listserver Archives
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Dear Fatima Merchant,

This is something that we can probably help you with. We do similar jobs
for several other companies.
If you could fax a drawing with the material, OD, Thickness, number of
holes, etc. to 1-802-878-8074 we can get you a quote.

Thanks,

John Arnott
Chairman
--

LADD RESEARCH
131 Dorset Lane
Williston, VT 05495

TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere)
FAX 1-802-878-8074
e-mail ladres-at-worldnet.att.net
web site http://www.ladd.cc


--

LADD RESEARCH
131 Dorset Lane
Williston, VT 05495

TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere)
FAX 1-802-878-8074
e-mail ladres-at-worldnet.att.net
web site http://www.ladd.cc






From: Fang Qian :      fangq-at-tr.comm.mot.com
Date: Wed, 23 Feb 2000 18:57:07 -0600
Subject: Cryotransfer system

Contents Retrieved from Microscopy Listserver Archives
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Hi, There,

We are considering to purchase a cryotransfer system for our 2010F TEM.
There are two major suppliers: Gatan and Oxford. We are not sure which
one works better since this is the first time dealing with this
technique.

Could you please send me your comments and experiences on these two
vendors? Our samples are mostly soft tissues: Polymers, surfactants,
cell membranes, and proteins. Your help is highly appreciated.

Maoxu Qian
****************************
* Maoxu Qian, Ph.D. *
* Dept of MSE, box 352120 *
* University of Washington *
* Seattle, WA 98195 *
* mxq-at-u.washington.edu *
* (206)616-3973(phone) *
* (206)543-3100(fax) *
****************************




From: David Henriks :      Henriks-at-CompuServe.COM
Date: Wed, 23 Feb 2000 19:31:13 -0600
Subject: Tripod Polisher Workshop

Contents Retrieved from Microscopy Listserver Archives
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Workshop Objective

This course will cover all aspects of pre-thinning and focus on final
thinning via Tripod Polishing. Due to the limited class size and the
extensive hands-on opportunities, this course is well suited to novices as
well as advanced Tripodders. Attendees will also learn the latest
techniques available in ion milling, plasma cleaning and ion beam sputter
deposition and etching for TEM and SEM samples.

The workshop will be held in San Clemente, CA on May 26-27, 2000. Please
call for more information or visit our web site ( www.southbaytech.com )
for registration information.


***************************************************************************

David Henriks
Vice President TEL: 800-728-2233
(toll free in the USA)
South Bay Technology, Inc. +1-949-492-2600
1120 Via Callejon FAX: +1-949-492-1499
San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com

***************************************************************************
} } } } } Please visit us at http://www.southbaytech.com { { { { {

Manufacturers of precision sample preparation equipment and supplies for
metallography, crystallography and electron microscopy.






From: Maoxu Qian :      mxq-at-u.washington.edu
Date: Wed, 23 Feb 2000 20:04:56 -0600
Subject: Cryo holder TEM

Contents Retrieved from Microscopy Listserver Archives
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}
}
} Hi, There,
}
} We are considering to purchase a cryotransfer system for our 2010F TEM.
} There are two major suppliers: Gatan and Oxford. We are not sure which one
} works better since this is the first time dealing with this technique.
} Could you please send me your comments and experiences on these two
} vendors? Our samples are mostly soft tissues: Polymers, surfactants, cell
} membranes, and proteins. Your help is highly appreciated.
}
} Maoxu Qian
} ****************************
} * Maoxu Qian, Ph.D. *
} * Dept of MSE, box 352120 *
} * University of Washington *
} * Seattle, WA 98195 *
} * mxq-at-u.washington.edu *
} * (206)616-3973(phone) *
} * (206)543-3100(fax) *
} ****************************
}
}
}
}
}






From: Divakar R :      divakar-at-igcar.ernet.in
Date: Thu, 24 Feb 2000 11:41:30 +0530
Subject: Re: Cool Prep (Water Coolers)

Contents Retrieved from Microscopy Listserver Archives
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A related question from India. We have been using DM (demineralised) water in our JEOL 2000 EX II cooled by a Neslab chiller for the last ten years. This requires change every 6 months approximately. What additives can be used to prevent algae (? it is green and slimy and I am not a biologist) growth? Something obtainable locally (chemical name rather than trade names) will be most useful to me. Thanks very much.

-----Original Message-----
} From: William Tivol [SMTP:tivol-at-wadsworth.org]
Sent: Thursday, February 24, 2000 10:25 AM
To: Robert Champaign
Cc: microscopy-at-sparc5.microscopy.com


} We are currently using Cool Prep in our Coolwell cooler system. This is a
} liquid cleaner & slime preventer that helps keep mineral deposits to a
} minimum in the cooling system. It also helps prevent corrosion in the
} water passages without harming hoses, gaskets or seals. Since Coolwell is
} no longer in business does anyone know of a replacement product for Cool
} Prep.
}
} Can we simply switch to ethylene glycol?

Dear Robert,
The suggestion of using distilled water and letting the system
come to equilibrium might work in a truly closed system composed of only
one material, but if you need to add water, or if you have several metals
in the system--as we do--this will be unsatisfactory in the long run.
We have found that Aqua-Treet 42 works very well in our Haskris system.
This product is made by Aqua Laboratories, Inc., PO Box 645, 8 Indus-
trial Way, Amesbury MA, (978) 388-3989. I have no relationship to this
company except as a satisfied user of their product.
Yours,
Bill Tivol







From: Anita Garg :      Anita.Garg-at-lerc.nasa.gov
Date: Thu, 24 Feb 2000 07:55:24 -0500
Subject: Re: Allied Multiprep Polisher for TEM samples

Contents Retrieved from Microscopy Listserver Archives
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Dear Colleagues
Does anybody have experience on the Allied MultiPrep Polishing
System, especially in terms of getting a good TEM sample? How well do
the Allied's Precision Cross-Section Tool and TEM Wedge Polisher work
on this semi automatic multiprep polisher?
Also, how does their TEM Wedge Polisher compare with the South Bay
Tripod Polisher?
TIA
Anita
*******************************************
Dr. Anita Garg
NASA Glenn Research Center at Lewis Field
Advanced Metallics Branch
Mail Stop 49-3
21000 Brookpark Road
Cleveland, OH 44135
Phone : (216) 433-8908
Fax : (216) 977-7132
E-mail : Anita.Garg-at-grc.nasa.gov
*******************************************




From: Dan May :      dmay-at-iti-med.com
Date: Thu, 24 Feb 2000 08:26:39 -0600
Subject: PINHOLE GRIDS

Contents Retrieved from Microscopy Listserver Archives
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Fatima Merchant,
I used to work for a company called Buckbee Mears. They have been making
tight tolerance electroformed Ni pinholes for many, many years. They are
located in St. Paul, MN and their phone number is 651-228-6400. Good luck.
Dan May






From: Leona Cohen-Gould :      lcgould-at-mail.med.cornell.edu
Date: Thu, 24 Feb 2000 10:47:41 -0400
Subject: Re: epoxy embedding of rat eye

Contents Retrieved from Microscopy Listserver Archives
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} Hi-
}
} We are going to be processing rat eyes (from perfused animals) for
} embedding in epoxy and I was wondering if anyone has any suggestions for
} processing. We are interested in the retina, optic nerve, cornea and uveal
} tract.
}
} Thank you very much,
} Sandy Perkins

****************************
Hi Sandra,

Eyes are a real pain. There are so many layers of different cell types,
and each seems to infiltrate differently. Having worked on a fir number of
eyes, I woauld make the following suggestions:
Process the cornea separately. It is almost pure collagen. It will not
osmicate all that noticeably, so don't be surprised when it doesn't turn
dark brown or black. Give it long infiltration times, in vaccuum if
possible.

If you don't need the optic nerve attached to the eye, dissect it free and
process it. Give it slighlty longer than usual dehydration and
infiltration steps to be sure you'e got the myelin taken care of.

As for the rest: remove the lens and the vitreous so that you have good
acess to the retina. Be careful not to disrupt the retina when you remove
the vitreous. Once the lens is out, you can usually get the vitreous out
by gently blotting against a Kimwipe. You may want to split the orb into
hemispheres. Just be sure not to take any sections from the cut edge,
since you will disturb the structure there when you cut. Again, you'll
nees longer dehyration and infiltration times, since the sclera is really
tough. I use 30 minutes in each step of graded acetone, propylene oc=xide
then a graded infiltraion with PO:resin (Epon analog) 2:1 for 30 min, 1:1
overnight, 1:2 2 hours, pure resin overnight, on a rotator.

I'm sure there ore others out there, with their own "pet protocol" for
eyes. Its all a bit of magic and luck, isn't it?

Good luck,
Lee

Leona Cohen-Gould, M.S.
Sr. Staff Associate
Director, Electron Microscopy Core Facility
Manager, Confocal Microscopy Core Facility
Joan & Sanford I. Weill Medical College
of Cornell University
voice (212)746-6146
fax (212)746-8175






From: ferrari-at-comp.uark.edu (Michael Ferrari)
Date: Thu, 24 Feb 2000 08:30:12 -0800
Subject: WI objectives for BH-2 RFCA

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Hi All,

Not sure this went out earlier...I've inherited an Olympus BH-2 RFCA
upright that I'd like to equip with WI lenses for live cell fluorescence
work. This is a 160mm tube length scope, and Olympus is now only
manufacturing the 60X WI fluor objective (I think/hope). Any leads to
sources of compatible WI objectives in the 10-60X range much appreciated.
Please reply offline. Thanks.

Mike

Michael Ferrari
Department of Biological Sciences
629 Science Engineering (SCEN)
University of Arkansas
Fayetteville, Arkansas 72701
Ph: 501-575-6372
Fax: 501-575-4010






From: amy-at-www.molbio.Vanderbilt.Edu (amy-at-helix.molbio.vanderbilt.edu)
Date: Thu, 24 Feb 2000 10:14:40 -0600
Subject: image processing - microdensitometer available

Contents Retrieved from Microscopy Listserver Archives
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We are interested in selling our Perkin-Elmer Model
1010M microdensitometer at a very reasonable price. It
was purchased as a reconditioned model from Perkin-Elmer
eight years ago and is in very good condition. Technical
manual is included.

If interested, please contact:

Amy Kendall
Department of Molecular Biology
Vanderbilt University
Box 1820, Station B
Nashville TN 37232
(615) 322-2012
amy-at-helix.molbio.vanderbilt.edu




From: Russell E. Cook :      recook-at-anl.gov
Date: Thu, 24 Feb 2000 11:03:00 -0600
Subject: RE: Cool Prep (Water Coolers)

Contents Retrieved from Microscopy Listserver Archives
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R. Divakar wrote, "A related question from India. We have been using DM
(demineralised) water in our JEOL 2000 EX II cooled by a Neslab chiller for
the last ten years. This requires change every 6 months approximately. What
additives can be used to prevent algae (? it is green and slimy and I am
not a biologist) growth? Something obtainable locally (chemical name rather
than trade names) will be most useful to me."

I quote here recommendations from the Haskris Co. (we have several Haskris
chillers):

1. If algae is present, flush the system with one of two alternatives: add
1 cup of chlorine bleach or one pint of hydrogen peroxide per 15 gallons of
water. Circulate 20-30 minutes. Drain the system and refill with clean
water.

2. To prevent regrowth of algae, one of the following additives are
recommended:
a. 10% solution of lab grade (no additives) ethylene glycol.
b. Dichlorophene. This is an insoluble white powder which is a
fungicide/bacteriacide. It should be sprinkled evenly across the entire
surface of the water in the chiller's tank.
c. 8 parts per million chlorine.

----------------------------------------------------------------------
Russell E. Cook, Ph.D.
Electron Microscopy Center for Materials Research
Argonne National Laboratory
Materials Science Division, Building 212
9700 South Cass Avenue
Argonne, IL 60439-4838
(630)252-7194
FAX: (630)252-4289
recook-at-anl.gov






From: Mary Mager :      mager-at-interchange.ubc.ca
Date: Thu, 24 Feb 2000 09:02:46 -0800
Subject: Re: (FE)SEM, sputter coating puzzle

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Dear Tobias,
The most logical explanation for this phenomenon is that the carbon-based
cell material is of a lower average atomic number, even with the thin Pt
coating, than the glass coverslip (SiO2) it is located on, also with the Pt
coat. The secondary electron yield is directly proportional to the average
atomic number, if topographic effects are not in evidence, which they won't
be on such a thin sample.
At 11:39 AM 2/23/00 -0600, you wrote:
} Greetings,
} I wonder if anyone could explain the following observation
} to me. I have been sputter coating with platinum and viewing the
} sample with a FESEM (Hitatchi 4700, below lens type). In a certain
} type of sample, the image appears darker than the background where
} there is sample. Now these samples are perhaps a bit unusual. They
} are very thin, starting off life as 1.75 um methacrylate sections of
} a plant root adhered to a glass coverslip, followed by extraction in
} hot acidic peroxide to remove all organic material except crystalline
} cellulose. The cellulosic cell walls left behind on the coverslip are
} nominally 100 nm thick and perhaps flattened further by the
} processing. The cell walls are present in the sample as either cross
} sections or as small regions laying in the plane of the coverslip.
} These regions can be anywhere from the size of a cell (10 x 40 or so
} microns) down to small portions thereof. All of this remaining cell
} wall is darker than the surrounding background. I am putting on a
} thin coat of Pt, somewhere around 2 nm, and imaging at 2 kV (I have
} seen the same thing at 5 kV). It really puzzles me that my sample is
} darker than background. Any explanations???
}
} THanks,
} Tobias

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





From: HILDEGARD CROWLEY :      hcrowley-at-du.edu
Date: Thu, 24 Feb 2000 10:43:06 -0700 (MST)
Subject: Re: Sudan Black staining and fixing

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Dear Toby,

You have a basic problem - free lipids interfere with the polymerization
of epoxies. Since you are having mixed results with osmium not all lipids
may be fixing adequately. If possible, try keeping the specimens in
osmium overnight, on a rotator. (After the first hour of exposure to
osmium, change to fresh osmium). I have done this very successfully with
lipid rich structures. Hope it works.

Hildy Crowley
University of Denver
Denver, CO





From: HILDEGARD CROWLEY :      hcrowley-at-du.edu
Date: Thu, 24 Feb 2000 10:53:58 -0700 (MST)
Subject: Re: Immuno-labeling of glutaraldehyd,

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On Wed, 23 Feb 2000, Dr. Manfred Rohde 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 Microscopist,
}
} has anyone out there any experiences with immuno-labeling of osmificated and
} Spurr embedded samples. We have tried out etching, oxidizing sections with
} hydrogen peroxide (10% for 15 min) or sodium methylat (1% for 15 min)
} without any significant increase of label intensity (Ab used is polyclonal
} from rabbit, protein A purified)
}
} Any suggestions are welcome. Manfred
}
}
}
Hello,

You may have a problem which you do not recognize. Spurr's really has no
place in immunocytochemistry. It is highly crosslinked. As you might
know, epoxies not only crosslink, they also chemically combine with tissue
proteins making them unavailable. Also, a highly stable, highly
crosslinked embeddding medium will cut a shiny, very smooth surface on
sections. The surface relief of such a section is extremely low, and, of
course, the more surface relief, the more chance for the antigen to be
exposed to markers. You might also have a paucity of antigen which
compounds all problems.

If you want to stay with epoxies, I would suggest switching to a soft
Epon-Araldite for starters. If that does not work, then my suggestion is
to go to LR Gold. A block of LR Gold does not "cut", but cleaves, causes
something like a triple increase in surface relief thus giving more
antigens the chance to be exposed.

If I am not making myself clear, please e-mail me. I have just gone
through 2yrs of frustration with stuff like this, but this week the paper
is going out!!!!

Bye,
Hildy Crowley
University of Denver
Denver, CO

P.S. I am not making all this up. I can reference everything, but do not
have the time to do that in this format





From: David Hoyle :      dhoyle-at-istar.ca
Date: Thu, 24 Feb 2000 13:22:52 -0800
Subject: Cooling water/glycol Danger

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Before adding glycol to your Haskris water chiller check with the
manufacture of the microscope. If you use glycol to cool a Hitachi
microscope you will damage the plastic water lines and they will all need
to be replaced after a few years. I have seen this happen a few times now
after people have used this advice from the list server. Please check with
your service engineer first.

To those users of Hitachi S2500/S570, glycol is used to cool the objective
lense but only on these 2 instruments.

I am a service engineer for Hitachi in Canada and the above opinion is my
opinion and not that of Hitachi.


David Hoyle
Nissei Sangyo Canada
http://www.nsctoronto.com/
service-at-nsctoroto.com (general mail)
dhoyle-at-istar.com (direct mail)




From: Heather A Owen :      owenha-at-csd.uwm.edu
Date: Thu, 24 Feb 2000 12:24:27 -0600 (CST)
Subject: RE: Cool Prep (Water Coolers)

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On Thu, 24 Feb 2000, Russell E. Cook wrote:

} I quote here recommendations from the Haskris Co. (we have several Haskris
} chillers):
}
} 1. If algae is present, flush the system with one of two alternatives: add
} 1 cup of chlorine bleach or one pint of hydrogen peroxide per 15 gallons of
} water. Circulate 20-30 minutes. Drain the system and refill with clean
} water.
}
} 2. To prevent regrowth of algae, one of the following additives are
} recommended:
} a. 10% solution of lab grade (no additives) ethylene glycol.
} b. Dichlorophene. This is an insoluble white powder which is a
} fungicide/bacteriacide. It should be sprinkled evenly across the entire
} surface of the water in the chiller's tank.
} c. 8 parts per million chlorine.
}
} ----------------------------------------------------------------------
} Russell E. Cook, Ph.D.
} Electron Microscopy Center for Materials Research
} Argonne National Laboratory
} Materials Science Division, Building 212
} 9700 South Cass Avenue
} Argonne, IL 60439-4838
} (630)252-7194
} FAX: (630)252-4289
} recook-at-anl.gov
}
}


I've been using dichlorophene in our Haskris chillers following the
incident related below. If you want to buy some, dichlorophene is listed
in several chemical company catalogs as:

2,2'-Methylenebis(4-chlorophenol)

Considering how I was planning to use it, I bought technical grade (90%),
since it was substantially less expensive than higher grades and have not
experienced any problems.

A complete replacement of our chiller water (along with the water in the
scope and lines) was required about a year ago when the water pump in our
TEM Haskris had to be replaced. Our sevice engineer flushed the system
using the concentrated hydrogen peroxide as mentioned above, and a truly
amazing amount of material spewed forth, so much so that small connectors
to the power supply clogged up a few weeks down the line, causing our HV
to shut down. Cleaning out the connectors and flushing the system again
with distilled water took care of the problem.

At the time, we were cautioned (by Haskris, I think) to use distilled
water and not deionized water in the chillers. Does anyone know why?

Heather Owen

Heather A. Owen, Director
Electron Microscope Laboratory
Department of Biological Sciences
University of Wisconsin - Milwaukee
(414)229-6816






From: Carl Henderson :      chender-at-umich.edu
Date: Thu, 24 Feb 2000 13:49:37 -0500
Subject: Re: Cooling water/glycol Danger

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OK, now I am concerned.

What is the real answer? When we had our Hitachi S-3200N installed
two years ago, the service engineer used an ethylene glycol/water
solution in the Haskris chiller. And it was pretty strong, too.
Something like 1:3 glycol:water.

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======================================
Carl Henderson
Electron Microbeam Analysis Laboratory
University of Michigan
2501 C.C. Little Bldg.
Ann Arbor, MI 48109-1063 USA
(734) 936-1550 FAX (734) 763-4690
======================================




From: Christensen, Kim :      ChristeK-at-whiteoaksemi.com
Date: Thu, 24 Feb 2000 14:38:10 -0500
Subject: Job Opening: TEM Technician

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TEM Technician
A position is available at White Oak Semiconductor, Richmond VA, USA. The
successful candidate will perform TEM operation and sample preparation in
support of DRAM fabrication and TEM engineering. The candidate should have
at least an associates degree and a minimum of 3 years experience in the
semiconductor industry; of which at least 1 year should pertain to
semiconductor TEM sample preparation. Additionally, they should understand
the fundamental tools and techniques of TEM sample preparation and the
semiconductor process from a top down and cross sectional perspective. Good
eye-hand coordination, communication skills, and attention to detail are
required. Candidates should be highly motivated, self directed, good at
problem solving, interested in learning new skills and effective working
alone or in a team environment.
Interested candidates please contact

Kim Christensen
Failure Analysis Laboratory
White Oak Semiconductor
6000 Technology Boulevard
Sandston, Virginia 23150
Ph: 804 952 7307
Fax: 804 952 7902
Pager: 1 800 759 8888, pin# 130 3382





From: Diana van Driel :      dianavd-at-eye.usyd.edu.au
Date: Fri, 25 Feb 2000 07:56:59 +1100
Subject: epoxy embedding of rat eye

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Sandy,

If you can puncture the eye, or preferably take off the anterior
chamber, so that the chemicals can get in, there is absolutely no
problem. Standard embedding, using longish times, works well. For
instance, 3 hrs in osmium, 1 hr in UAc, 20 mins in each of the
alcohols/acetone, 1 hr in resin/acetone (dilute), overnight
infiltration in resin/acetone (more concentrated), 1 hr in warm
resin, embed. If you need to have the eye unpunctured, it may be
difficult. I've never embedded a discrete eye, but I do know that it
is really difficult to get even isolated sclera well embedded, as it
seems to act as a very effective barrier to resin. The eye is a well
designed ball that doesn't want to deflate, but that also means
nothing much will get in.

Contact me if you want further info. I've embedded more eyes than I
care to remember, including some rats eyes.

Diana



Diana van Driel
Department of Clinical Ophthalmology
Sydney University
GPO Box 4337
Sydney NSW
AUSTRALIA 2001

Phone 61 2 938 27278/27395
Mob 0412 165 075
Fax 61 2 938 27318




From: David Hoyle :      dhoyle-at-istar.ca
Date: Thu, 24 Feb 2000 16:15:16 -0800
Subject: Cooling water/glycol Danger

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Carl,

You are correct Carl you can use glycol to cool your sem(all Hitachi sems)
as only the diffusion pump is being cooled and Hitachi uses a vinyl hose
that isn't effected by the Glycol. For those that have Hitachi TEM's the
water lines for the lenses and diffusion pumps are the smaller black
appearing lines, these will be adversely effected by the glycol.

Sorry for the confusion I will be more specific in the future. So in the
sem's that use the clear reinforced vinyl hoses only, glycol is okay. In
Hitachi TEM which use a small black plastic hose for the lenses and
diffusion pumps, do not use glycol.



} OK, now I am concerned.

} What is the real answer? When we had our Hitachi S-3200N installed
} two years ago, the service engineer used an ethylene glycol/water
} solution in the Haskris chiller. And it was pretty strong, too.
} Something like 1:3 glycol:water.

David Hoyle
Nissei Sangyo Canada
http://www.nsctoronto.com/
service-at-nsctoroto.com (general mail)
dhoyle-at-istar.com (direct mail)




From: Brian Wajdyk :      r49655-at-email.sps.mot.com
Date: Thu, 24 Feb 2000 14:39:06 -0700
Subject: Job announcement: TEM technician

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I am pleased to bring to your attention the following TEM technician
position at Motorola's Process and Materials Characterization Laboratory
(PMCL) I encourage all persons interested to respond ASAP.

Position: TEM Technician/S9001P
Employer: Motorola-Semiconductor Products Sector's Process and Materials
Characterization Laboratory (PMCL)
Location: Mesa, AZ
Employment type: Full-time
Employment status: Full-employee (non-contractor)
Number of Positions: 2
Shift: all (compressed)
Relocation: Available

Duties/Responsibilities: Experience in Transmission Electron Microscopy
techniques including basic operation, specimen preparation, maintenance,
and troubleshooting of TEM's. Work effectively as a team player in
multiple projects providing routine and non-routine TEM analysis in
support of semiconductor product manufacturing. Exposure to many
different types of processes and technologies, and working knowledge of
FIB and EDS are a plus.

Specific knowledge: AA degree preferred. A higher or lower
classification will be established depending upon qualification and
experience.

Contact info: Send resume or questions via email to
Brian_Wajdyk-at-email.mot.com or fax to 480-655-4316 C/O Brian Wajdyk.
--
********************************************************************
Brian Wajdyk
Team Leader / Electron Microscopist (FESEM, EDS, SAM)
Motorola - Process and Materials Characterization Laboratory (PMCL)
2200 W. Broadway Rd., Mesa AZ 85202 Mail Drop: M360
Tel: 480-655-4337 Fax: 480-655-4316
Email: brian_wajdyk-at-email.mot.com Pager: 1-800-313-5960
********************************************************************




From: DakruegerMN1-at-aol.com
Date: Thu, 24 Feb 2000 16:49:33 EST
Subject: image analysis of fibers

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Image analysis folks,

I am interested in doing some PC based image analysis of fibers and fiber
properties on crimping. Is there anyone doing IA of crimp angle/amplitude or
crimps per inch? Is there commercial IA software that will do these types of
measurements?

Any help would be appreciated. Thanks.

Duane Krueger
7635 Harold Avenue
Golden Valley, Minnesota 55427

612-541-9880 (FAX)
DakruegerMN1-at-aol.com
yanktonson2-at-aol.com




From: William Tivol :      tivol-at-wadsworth.org
Date: Thu, 24 Feb 2000 18:00:36 -0500
Subject: Re: Cool Prep (Water Coolers)

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Gordon Couger wrote:

} Glycols won't have near the heat capacity of water. I doubt
} you will find a bug that will live in ethylene glycol but you can
} probably find many that can live in propylene glycol.
}

Dear Gordon,
I've been trying to look up the heat capacity of glycols
in my Handbook of Chemistry and Physics. It doesn't have the
data I want, but it does have the Cp for water (~4 j/gK), butane
(~0.006 j/molK = ~10^-4 j/gK, if I'm reading my cal's and
Cal's right), and 1-propanol (~0.01 j/molK with the same caviat).
} From these, I'd guess Cp for ethylene glycol is ~0.014 j/molK,
which is, indeed, much less than that of water. Propylene
glycol's Cp should be similar.
Yours,
Bill Tivol





From: William Tivol :      tivol-at-wadsworth.org
Date: Thu, 24 Feb 2000 18:08:15 -0500
Subject: Re: Cool Prep (Water Coolers)

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Divakar R wrote:

} A related question from India. We have been using DM (demineralised) water in our JEOL 2000 EX II cooled by a Neslab chiller for the last ten years. This requires change every 6 months approximately. What additives can be used to prevent algae (? it is green and slimy and I am not a biologist) growth? Something obtainable locally (chemical name rather than trade names) will be most useful to me. Thanks very much.

Dear Divakar,
Russell's suggestion of dichlorophene is good--it's also
what we use. To prevent growth in the tubing, we added some
CuSO4--being careful to keep the pH slightly above 7.5.
We have copper tubing in the system, so the Cu++ should not
be a problem, and it will inhibit algal growth. At that pH,
CuSO4 is not too soluble, so go slowly (if at all).
Yours,
Bill Tivol





From: William Tivol :      tivol-at-wadsworth.org
Date: Thu, 24 Feb 2000 18:15:47 -0500
Subject: Re: Cool Prep (Water Coolers)

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Heather A Owen wrote:

Dear Heather,


} A complete replacement of our chiller water (along with the water in the
} scope and lines) was required about a year ago when the water pump in our
} TEM Haskris had to be replaced. Our sevice engineer flushed the system
} using the concentrated hydrogen peroxide as mentioned above, and a truly
} amazing amount of material spewed forth, so much so that small connectors
} to the power supply clogged up a few weeks down the line, causing our HV
} to shut down. Cleaning out the connectors and flushing the system again
} with distilled water took care of the problem.
}
}

We added inline filters--the ones with the fiber cartridges--to
trap all the particulate material in the lines, and over a period of several
months, we probably got as much material as you got all at once. I highly
recommend adding filters.

At the time, we were cautioned (by Haskris, I think) to use distilled

} water and not deionized water in the chillers. Does anyone know why?
}

Hard to say. Deionized water can contain material leached out
of the resin used, and maybe that is bad for the system, while volatiles
(mostly organics, I'd guess) from distillation may not be.
Yours,
Bill Tivol





From: Andrew McNaughton :      andrew.mcnaughton-at-stonebow.otago.ac.nz
Date: Fri, 25 Feb 2000 12:31:14 +1300
Subject: Reference points in serial sections.

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Hello

A student wishes to take a number of serial sections through a bee brain
mounted in glycerine. The total distance is around 500m and she would
like to include three reference points which enable the sections to be
correctly orientated throughout the brain. Embedding three small rods
seems to be the most likely option but the material would need to be stable
and able to be cut by a vibratome. Keeping the rods parallel would also be
beneficial.

If anyone has tried this sort of thing or has any suggestions as to what we
could use for the markers I'd appreciate hearing your bright ideas.

Regards

Andrew McNaughton

______________________________________________________________________________
Andrew McNaughton
South Campus Electron Microscope Unit
C/-Department of Anatomy and Structural Biology
School of Medical Sciences
University of Otago
PO Box 913, Dunedin
NEW ZEALAND

Telephone: +64 3 479 7308
Facsimile: +64 3 479 7254

e-mail: andrew.mcnaughton-at-stonebow.otago.ac.nz
______________________________________________________________________________ht






From: Hkan Olin :      hakan.olin-at-fy.chalmers.se
Date: Fri, 25 Feb 2000 08:05:38 +0100
Subject: SPM information on the Web - including TEM-STM

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Dear Timothy,

We are using sputter coater in "etch" mode for this purpose. It is doing the
job flawlessly.
I don't know about your coater but I'll give you the settings we are using
here.
The coater has maximum operating voltage of 1400V. The grids are put on
glass plate (so they do not make contact with the lower electrode) and then
the plate is put on the lower electrode. The current we use is 5 mA for 20
seconds (the current depends strongly on the pressure and the voltage). The
pressure is about 20 Pa.

Good luck !

Rado

---------------------------------------------------------------------
Radostin Danev
Laboratory of Ultrastructure Research
National Institute for Physiological Sciences
Myodaiji-cho, Okazaki 444-8585, JAPAN
e-mail: rado-at-nips.ac.jp
---------------------------------------------------------------------
----- Original Message -----
} From: Timothy Schneider {Timothy.Schneider-at-mail.tju.edu}
To: {microscopy-at-sparc5.microscopy.com}
Sent: Thursday, February 24, 2000 4:29 AM


Dear Microscopy users,

I just want to inform you about a new web site
(http://www.nanofactory.com) with some useful information on scanning
probe microscopy (SPM)

There are, for example:
- Weekly update of SPM research articles
- Suggestions of some SPM literature
- Links to many SPM research groups
- Links to almost all SPM companies

There are also some interesting mpeg-movies with TEM-STM measurements of
two tunneling tips jumping into each other.

Best regards,
Hakan Olin
--
Dr Hakan Olin
Physics and Engineering Physics
Chalmers University of Technology
SE-412 96 Goteborg
Sweden
tel. +46-31-772 3338
fax +46-31-772 3367
mobile +46-70-30 88 99 0
e-mail hakan.olin-at-fy.chalmers.se
home page http://fy.chalmers.se/~f4aolin




From: Wim Jacob :      jacob-at-uia.ua.ac.be
Date: Fri, 25 Feb 2000 11:31:08 +0100
Subject: Re: Food Science Prep.

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An effective way to prevent your cooling system from growing
green or blue-green algae is to ensure that it is totally light-tight.
Algae require light for photosynthesis. The Philips engineers who
installed our CM120 Biotwin recommended we connect up the
chiller with the black-lined vinyl hose used in drinks vending
machines, where microorganism growth in the water lines is
obviously undesirable. The product we used is trademarked
"Aquavend", is opaque white outside and black inside. It appears to
have worked so far, though the ethylene glycol we use would also
discourage algae.
Yours
Chris Jeffree

Date sent: Thu, 24 Feb 2000 18:08:15 -0500
} From: William Tivol {tivol-at-wadsworth.org}
To: microscopy-at-sparc5.microscopy.com


Dear Debby,
Some ten yaers ago we were doing research on sugar syrup in cooperation with a sugar company. They were interested in the stucture of mixtures of sugar syrups. W e had some succes using freeze-fracture and looking at the replicas in the TEM. Of course these were not mixtures of cereals with sugar syrup and the specimens were allowed to be a lot smaller but this could be a suggestion.
Succes !
Wim Jacob
Ctr.of Electron Microscopy
University of Antwerp
Belgium

Debby Sherman wrote:

} ------------------------------------------------------------------------
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} -----------------------------------------------------------------------.
}
} Hi,
} Well I guess we never run out of new challenges when it comes to specimen prep. This one concerns a product that is made of a cereal bound together with a sugar syrup. The investigators want to see how the syrup behaves as to structural integrety over a period of time with and without the addition of various stabilizing components. Information would be collected at the light microscopic level.
} We first thought of cryosectioning followed by examination using LM. However, the product is filled with air pockets and literally disintegrates when microtomed. It is easily cut into pieces at temperatures above freezing as long as the pieces are no thinner than3-4 mmAccempts to infiltrate it with OCT compoound prior to freezing were not satisfactory.
} New thought is to use a low temperature resin to infiltrate the sample followed by UV or chemical polymerization. butThe resulting blocks could then be microtomed at room temperature and imaged. It would be desirable to cut sections of 4-8 m in thickness which is too thick for an ultramicrotome with glass or diamond knives. Also UV polymerization would probably limit us to very small sample size which may be a problem.
} Any suggestions would be appreciated as to resins to use which have low viscosity, could be used to infiltrate pieces of ~0.5-1cm sq, polymerize at low temperatures (+4 to 0oC range) using UV or chemical polymerization, and are preferably soft enough to section with a metal microtome blade. Any other suggestions of preparations techniques to try would be very appreciated.
}
} Thanks in advance for your input.
} Debby
}
} Debby Sherman, Manager Phone: 765-494-6666
} Microscopy Center in Agriculture FAX: 765-494-5896
} Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu
} Purdue University
} 1057 Whistler Building
} West Lafayette, IN 47907-1057





From: Wim Jacob :      jacob-at-uia.ua.ac.be
Date: Fri, 25 Feb 2000 11:52:17 +0100
Subject: Re: Can EELS, WDS or FT-IR localize As in Biol. tissues?

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Dear Doug,
Why don't you try imaging SIMS? I would not surprised if such instrument(s) are
present at the University of Arizona.
Of course this would restrict the resolution, I gess, to 0.1 - 0.5 micrometer,
with the newer instruments.
Regards
Wim Jacob
Ctr. of Electron Microscopy
University of Antwerp
Belgium

Doug Cromey wrote:

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} -----------------------------------------------------------------------.
}
} Colleagues,
}
} We are interested in localizing arsenic in biological tissues (toxicology
} research), but have been unable to come up with a technique that will give
} us the cellular localization information we're looking for (sub-cellular
} would be even better).
}
} We have tried TEM with energy dispersive x-ray microanalysis and our
} concentrations are apparently too low, so we are unable to detect arsenic.
}
} A few years ago we looked into using the autometallographic techniques that
} have been published by Dr. Gorm Dansher & his colleagues. We were told
} that the photographic emulsions required were no longer available.
}
} In a brainstorming session yesterday several techniques were bantered
} about, but in truth we are not familiar enough with the techniques to know
} if they would suit our needs. Does EELS have anything to offer, or perhaps
} wavelength dispersive spectoscopy (WDS)? How about some of the new FT-IR
} instruments? Are there any other techniques that might be able to localize
} arsenic and/or indicate its state of oxidation?
}
} Vendors are welcome to reply.
}
} Yours,
} Doug Cromey
} ....................................................................
} : 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: doug-cromey-at-ns.arizona.edu):
} :...................................................................:
} http://www.pharmacy.arizona.edu/exp_path.html
} Home of: "Microscopy and Imaging Resources on the WWW"





From: Shu-You Li :      syli-at-mail.uni-mainz.de
Date: Fri, 25 Feb 2000 15:56:01 +0100
Subject: TEM: a new TEM website.

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Hi, all,

Please visit my new homepage at http://syli.homepage.com at your convinience!
It contains not only my current research work, but also LINKS to useful journal informations, instruction for authors, on-line EELS database, periodic table, physical constants, online TEM course and etc.

I am looking forward to your invaluable suggestions!
Yours sincerely,

Shu-You Li


**************************************************
Shu-You Li, Dr.
Institut fuer Physikalische Chemie
Johannes Guttenberg Universitaet
Jakob-Welder-Weg 11
D-55099 Mainz, Germany

E-mail: syli-at-mail.uni-mainz.de; syli16-at-hotmail.com
Fax: +49-6131-3923768
Tel: +49-6131-3923148(O)
**************************************************








From: Jill Verlander :      verlaj-at-medicine.ufl.edu
Date: Fri, 25 Feb 2000 14:25:57 -0500
Subject: Immunogold Workshop

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Becky,
I do not know if it is okay in the S4700 to use glycol. If it is working
as in why change?


X-From_: r-holdford-at-ti.com Thu Feb 24 19:53:24 2000


X-From_: Patrick.Kilcran-at-hii-hitachi.com Thu Feb 24 23:55:28 2000
} From: "Kilcran, Patrick-HIIEX" {Patrick.Kilcran-at-hii-hitachi.com}
To: dhoyle-at-istar.ca
Cc: "Lima, Rick-HIIEX" {Rick.Lima-at-hii-hitachi.com}


Dear Researcher:

The University of Florida College of Medicine Electron Microscopy Facility
is hosting a two-day workshop on immunogold technique. The workshop will
include two identical sessions on April 10/11 and 13/14. Dr. Jan
Leunissen from the Aurion Immunogold Reagent & Accessories, an
internationally known expert in the field, will be the instructor for the
workshop. Attached is the program information. If you are interested in
attending this workshop, please contact Dr. Verlander or Ms. Hong Yi at
the addresses provided below. Due to practical considerations, the
workshop will be limited to a maximum of 20 participants for each session.
Therefore, we encourage you to register as soon as possible. If you are
not able to attend, we would appreciate if you could pass the message onto
someone else who might be interested in learning about immunogold
techniques. Thank you very much.

Dr. Jill W. Verlander Reed (workshop faculty)
Director, University of Florida College of Medicine Electron Microscopy
Facility Phone: (352) 846-0820 Fax: (352) 846-3299 verlaj-at-medicine.ufl.edu

Hong Yi (technical coordinator)
Supervisor, Emory Neurology Microscopy Core Laboratory
Phone: (404) 727-8692
Fax: (404) 727-3157
Email: hyi-at-emory.edu


University of Florida, College of Medicine, Electron Microscopy Facility
Aurion Immunogold Reagents & Accessories / Electron Microscopy Sciences

Present
Immunogold Workshop

April 10-14, 2000
Gainesville, FL

WORKSHOP HOST
Dr. Jill Verlander Reed
Director, Electron Microscopy Facility
University of Florida College of Medicine
Phone: (352) 846-0820
Fax: (352) 846-3299
verlaj-at-medicine.ufl.edu

WORKSHOP INSTRUCTORS

Dr. Jan L.M. Leunissen is an internationally known scientist in the field
of ultrastructural localization. He received his Ph.D in molecular cell
biology at the University of Utrecht in the Netherlands. Dr. Leunissen is
the inventor of ultra small colloidal gold probes and also the founder and
president of the company Aurion, which specializes in immunogold reagents
and custom immunogold labeling. He has been invited to many international
microscopy conferences and workshops and is especially experienced in
providing hands-on training. His previous workshops in Europe, Asia, and
the US have been fully attended and very well received.

Dr. Jill W. Verlander is the Director of the Electron Microscopy Facility
for the University of Florida College of Medicine. Dr. Verlander has 15
years of experience in ultrastructural research in kidney and is known
internationally for her work examining structure-function relationships in
renal transport epithelia. Her work has been published in such journals as
the Journal of Clinical Investigation, the American Journal of Physiology,
Journal of the American Society of Nephrology, and Kidney International.
These studies have been accomplished using light microscopic
immunohistochemistry and in situ hybridization, scanning and transmission
electron microscopy, morphometric analyses, and immunogold and
immunoperoxidase cytochemistry at the ultrastructural level.

GENERAL INFORMATION
Objectives
To provide researchers the opportunity to learn the theory and practice of
immunogold labeling To permit participants to process their own samples
using these techniques under expert guidance To promote technology
exchange and research collaboration

Registration Fees
Regular: $250.00
Student: * $150.00

Lodging
Sheraton Gainesville, $69/night
Phone: (352) 373-6721

Rooms will be reserved in the above hotel under the name "UF EM workshop".
Shuttle buses are available between the hotel and UF campus

Enrollment Note
Two sessions (A; April 10-11, B: April 13-14) will be hosted at UF. Each
session will be limited to a maximum of 20 participants. The workshops
will provide samples to those who prefer not to bring their own. Each
participant will be provided with a sample of gold conjugate of their
choice at the end of the workshop.

MAIN CURRICULUM
The properties of gold particles and their protein conjugates.
Theories underlying immunogold labeling protocols.
Silver enhancement of gold particles
Imunogold labeling on a variety of sample preparations for LM.
Immunogold labeling for EM
a. Pre-embedding immunogold labeling using ultra_small gold conjugates
and silver enhancement. b. Post-embedding immunogold labeling using ultra
small and conventional colloidal gold conjugates.

CONTACT PERSON AND TECHNICAL COORDINATOR
Hong Yi
Phone: (404) 727-8692
Fax: (404) 727-3157
Email: hyi-at-emory.edu


Jill Verlander Reed, D.V.M.
Associate Scientist
Office of the Dean
University of Florida College of Medicine
P.O. Box 100215
Gainesville, FL 32610
verlaj-at-medicine.ufl.edu
Phone: (352) 846-0820
Fax: (352) 846-3299




From: george sibbald :      geos-at-goldrush.com
Date: Fri, 25 Feb 2000 13:53:49 -0700
Subject: AFM Newslatter

Contents Retrieved from Microscopy Listserver Archives
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Molecular Imaging is updating our contact information.

If you are interested in scanning probe microscopy, would you please take a
moment and update your information at http://www.molec.com/info/form.html

Thank you

George

_______________________
George Sibbald, President
Molecular Imaging: Technology Leader In Advanced Bio-AFM Systems
9830 South 51 Street, Ste 124A
Phoenix AZ 85044
(480) 753-4311
www.molec.com






From: Delilah Wood :      wood-at-pw.usda.gov
Date: Fri, 25 Feb 2000 14:03:54 -0800
Subject: Re: Glycol for cooling

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In fact, we have an S-4700 and the service technician told us that there are tons of problems with using glycol in the chillers. We have a Haskris chiller. Apparently, the glycol volatilizes some of the hose components.


De Wood


At 11:01 AM 2/25/2000 -0800, David Hoyle wrote:

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} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html

} -----------------------------------------------------------------------.

}

}

} Becky,

} I do not know if it is okay in the S4700 to use glycol. If it is working

} as in why change?

}

}

} X-From_: r-holdford-at-ti.com Thu Feb 24 19:53:24 2000

} Date: Thu, 24 Feb 2000 19:26:10 -0600

} } From: Becky Holdford { {r-holdford-at-ti.com}

} Organization: KFAB Physical Analysis Lab, Texas Instruments, Dallas, TX

} X-Mailer: Mozilla 4.51 [en] (WinNT; U)

} X-Accept-Language: en,en-US,en-GB,uk,ru

} To: David Hoyle { {dhoyle-at-istar.ca}

} Subject: Re: Cooling water/glycol Danger

} X-MIME-Autoconverted: from 8bit to quoted-printable by tower.ti.com id

} TAA19583

}

} David: we have 3 Hitachi S4700's that have the final lenses water-cooled.

} Does the caveat of glycol still hold? We are using distilled water and have

} had no problem so far.

}

} David Hoyle wrote:

}

} } ----------------------------------------

}

}

{bold} ********************************************************************

Delilah F. Wood

Botanist

USDA-ARS-WRRC

800 Buchanan St.

Albany, CA 94710


Tel: 510-559-5653

Fax: 510-559-5818 {/bold}

{/x-rich}



From: gregory.argentieri-at-pharma.Novartis.com
Date: Fri, 25 Feb 2000 18:08:34 -0600
Subject: Phtographic printers for TEM

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}
} Looking to find and to determine what is the best printer (photographic and
} archive quality)
} for TEM digital images.
}
} Currently considering Kodak 8670PS, Codonics, or Fuji Printers
} Would be interested in comments from people using photoraphic qualtiy
} printers
} for TEM applications
} thoughts, "next time's" etc.
}
} Thank You
}
} Gregory Argentieri
} Novartis Pharmacuticals Corp
} gregory.argentieri-at-pharma.novartis.com
}
}
}
}






From: Purdy, Sam :      SPurdy-at-nationalsteel.com
Date: Fri, 25 Feb 2000 18:10:57 -0600
Subject: RE: Food Science Prep.

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Debby Sherman
Talk to Mark Cavaleri at 3M. He made a polished cross sectgion of M%M
candies and preserved tyhe sugr layer

Sam Purdy

} ----------
} From: Debby Sherman
} Sent: February 2000 1:17 PM
} To: message to: MSA list
} Subject: Food Science Prep.
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
} -----------------------------------------------------------------------.
}
}
} Hi,
} Well I guess we never run out of new challenges when it comes to
} specimen prep. This one concerns a product that is made of a cereal bound
} together with a sugar syrup. The investigators want to see how the syrup
} behaves as to structural integrety over a period of time with and without
} the addition of various stabilizing components. Information would be
} collected at the light microscopic level.
} We first thought of cryosectioning followed by examination using LM.
} However, the product is filled with air pockets and literally
} disintegrates when microtomed. It is easily cut into pieces at
} temperatures above freezing as long as the pieces are no thinner than3-4
} mmAccempts to infiltrate it with OCT compoound prior to freezing were not
} satisfactory.
} New thought is to use a low temperature resin to infiltrate the sample
} followed by UV or chemical polymerization. butThe resulting blocks could
} then be microtomed at room temperature and imaged. It would be desirable
} to cut sections of 4-8 m in thickness which is too thick for an
} ultramicrotome with glass or diamond knives. Also UV polymerization would
} probably limit us to very small sample size which may be a problem.
} Any suggestions would be appreciated as to resins to use which have low
} viscosity, could be used to infiltrate pieces of ~0.5-1cm sq, polymerize
} at low temperatures (+4 to 0oC range) using UV or chemical polymerization,
} and are preferably soft enough to section with a metal microtome blade.
} Any other suggestions of preparations techniques to try would be very
} appreciated.
}
} Thanks in advance for your input.
} Debby
}
} Debby Sherman, Manager Phone: 765-494-6666
} Microscopy Center in Agriculture FAX: 765-494-5896
} Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu
} Purdue University
} 1057 Whistler Building
} West Lafayette, IN 47907-1057
}
}






From: Mei Lie Wong :      wong-at-msg.ucsf.edu
Date: Fri, 25 Feb 2000 16:10:42 -0800
Subject: Fwd: sem

Contents Retrieved from Microscopy Listserver Archives
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I received this today. If anyone could help, please reply to the sender.

Thanks.

ML

} Date: Fri, 25 Feb 2000 17:14:04 -0600
} From: Howard Hsu {hhsu-at-kumc.edu}
} Organization: KU Medical Center
} X-Accept-Language: en
} MIME-Version: 1.0
} To: wong-at-msg.ucsf.edu
} Subject: sem
}
} I am a researcher at the University of Kansas Med. Center and would like
} study a biological sample for the presence of mineral using SEM
} approach. If you can provide external service on cost or collaborative
} basis, please let me know. Otherwise, I would greatly appreciate if you
} would let me know any academic or industrial sources that provide this
} type of service. Thank you. Howard Hsu, Ph.D.
}
Mei Lie Wong
Electron Microscope Laboratory
Department of Biochemistry
HHMI-UCSF
Ph. 415-476-4441 Fax 415-476-1902
http://util.ucsf.edu/agard/wong/index.html
email wong-at-msg.ucsf.edu




From: Ronald R. Matias :      rrm-at-skyinet.net
Date: Sat, 26 Feb 2000 08:19:49 +0800
Subject: Need help on EM negative staining

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html



Hello and Help!

We are a hospital based research lab. starting to use the electron
microscope to identify or confirm pathogens that cause gastrointestinal
disturbances. We would like to know if someone out there could recommend
appropriate specific EM protocols (negative staining, processing of the
gastrointestinal contents, etc....) to visualize these pathogens (ranging
from viruses to bacteria and protozoans). We have a Jeol 1010 EM and perform
the routine EM preparations designed to examine tissues.

In the interest of bring quality healthcare to our people, we will be very
much obliged,

ronnie matias




From: Garber, Charles A. :      cgarber-at-2spi.com
Date: Sat, 26 Feb 2000 05:13:31 -0500
Subject: SEM/TEM analytical service request

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --

Mei Lie Wong wrote:
==================================================
I received this today. If anyone could help, please reply to the sender.

Thanks.

ML

} Date: Fri, 25 Feb 2000 17:14:04 -0600
} From: Howard Hsu {hhsu-at-kumc.edu}
} Organization: KU Medical Center
} X-Accept-Language: en
} MIME-Version: 1.0
} To: wong-at-msg.ucsf.edu
} Subject: sem
}
} I am a researcher at the University of Kansas Med. Center and would like
} study a biological sample for the presence of mineral using SEM
} approach. If you can provide external service on cost or collaborative
} basis, please let me know. Otherwise, I would greatly appreciate if you
} would let me know any academic or industrial sources that provide this
} type of service. Thank you. Howard Hsu, Ph.D.
}
Mei Lie Wong
Electron Microscope Laboratory
Department of Biochemistry
HHMI-UCSF
Ph. 415-476-4441 Fax 415-476-1902
http://util.ucsf.edu/agard/wong/index.html
email wong-at-msg.ucsf.edu
============================================================
Although you have specifically asked about SEM, I am aware only of a TEM
approach for this kind of analysis. We at Structure Probe, Inc. do perform
this kind of work for clients on a commercial basis. You can see an example
of the final sample that is analyzed by TEM on our website at page URL
http://www.2spi.com/catalog/instruments/etchers4.html

The method is pretty much described so anyone really could do with, provided
they did have access by a plasma etcher such as the SPI Plasma Prep II. If
you wanted to do it yourself and were having difficulties, let us know,
perhaps we could give you assistance. We have found that the removal of the
organics is usually quite important in order to remove the source of the
unwanted Bremsstrahlung radiation.

We have heard from persons trying to do this by SEM/EDS, where by the
section is deposited on a polished beryllium mount, and then the organics
etched away but they seem to have not been successful.

Disclaimer: SPI Supplies manufactures the plasma etching equipment needed
to practice this procedure and our Structure Probe analytical services part
of our business performs this kind of sample preparation and analysis for
clients. We realize there might be other methods for doing this, but we
have favored the approach that works for us.

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.spi.cc
########################
============================================






From: przybylo :      przybylowicz-at-nac.ac.za
Date: Sat, 26 Feb 2000 22:44:21 +0200
Subject: As in biological tissues

Contents Retrieved from Microscopy Listserver Archives
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Dear Doug,


We have some experience in using micro-PIXE (proton induced X-ray emission)
and the so-called nuclear microprobe in toxicological research. It is more
sensitive than EDS and WDS, is very easy to quantify (standardless
technique). In principle you can have detection limits of the order of 1
ppm in point analysis mode. Specimens can be of any thickness and their
actual thickness is assessed by the simultaneous use of the BS technique
(proton backscatering spectrometry). What in my opinion makes it really
attractive for this purpose is its very good scanning capability. Scan sizes
are from ca. 2.5 mm x 2.5 mm down to the beam resolution, which is of the
order of few micrometer (can be down to 1 micrometer or even less, but in
most of our uses few micrometers is good enough).
The sample preparation is similar as for EDS, using cryotechniques to avoid
elemental losses and redistribution.
We have so far done studies on Ni, Mo, Cd. The detection limits for As are
better than for Mo and Cd.
Of course, this technique cannot indicate the state of oxidation, you can
really treat it as a "more sensitive EDS".

If you want more information, I can refer you to our earlier work and
perhaps you might be interested in contacting me.

Best regards

xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
Dr Wojciech J. Przybylowicz
Materials Research Group
National Accelerator Centre
PO Box 72
Faure 7131 South Africa
E-mail: przybylowicz-at-nac.ac.za
Fax: +27-21-8433543
Phone: +27-21-8433820
http://www.nac.ac.za/IBA/
http://www.nac.ac.za/materials/
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx





From: przybylo :      przybylowicz-at-nac.ac.za
Date: Sat, 26 Feb 2000 22:50:42 +0200
Subject: As in biological tissues

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Dear Doug,


We have some experience in using micro-PIXE (proton induced X-ray emission)
and the so-called nuclear microprobe in toxicological research. It is more
sensitive than EDS and WDS, is very easy to quantify. In principle you can
have detection limits of the order of 1 ppm in point analysis mode.
Specimens can be of any thickness and their actual thickness is assessed by
the simultaneous use of the BS technique (proton backscatering
spectrometry). What in my opinion makes it really attractive for this
purpose is its very good scanning capability. Scan sizes are from ca. 2.5 mm
x 2.5 mm down to the beam resolution, which is of the order of few
micrometer (can be down to 1 micrometer or even less, but in most of our
uses few micrometers is good enough).
The sample preparation is similar as for EDS, using cryotechniques to avoid
elemental losses and redistribution.
We have so far done studies on Ni, Mo, Cd. The detection limits for As are
better than for Mo and Cd.
Of course, this technique cannot indicate the state of oxidation, you can
really treat it as a "more sensitive EDS".

If you want more information, I can refer you to our earlier work and
perhaps you might be interested in contacting me.

Best regards

xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
Dr Wojciech J. Przybylowicz
Materials Research Group
National Accelerator Centre
PO Box 72
Faure 7131 South Africa
E-mail: przybylowicz-at-nac.ac.za
Fax: +27-21-8433543
Phone: +27-21-8433820
http://www.nac.ac.za/IBA/
http://www.nac.ac.za/materials/
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx





From: John Catino :      jwcatino-at-concentric.net
Date: Sun, 27 Feb 2000 21:34:39 -0500
Subject: Print Processor for sale

Contents Retrieved from Microscopy Listserver Archives
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I have a Kodak Royal Print Processor for sale. The unit is in good
working condition. Our lab is going digital.

Please contact me via e-mail if interested.

John







From: John Catino :      jwcatino-at-concentric.net
Date: Sun, 27 Feb 2000 21:25:36 -0500
Subject: Kodak Royal Print Processor

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


I have a Kodak Royal Print Processor for sale. The unit is in good
working condition. Our lab is going digital.

Please contact me via e-mail if interested.

John





From: Hans-Werner Liebsch :      hwl.bioanalytic-at-bluewin.de
Date: Mon, 28 Feb 2000 11:22:52 +0100
Subject: Active Vibration Isolation News

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An information about a new principle in vibration damping to all
microscopy users, who have problems with particularly low frequency
vibrations from natural environment.

I invite you kindly to visit our home page:
http://www.hwlbioanalytic.com

We show there active vibration isolation systems for various
applications with damping frequencies from 0,6 Hz to 100/200 Hz and load
capacities (depending from system) from max. 90 kg up to 330/660 kg and
max. 1.500 kg.
Thank you for your time.

With kind regards
Hans-Werner Liebsch

--------------------------------------------
HWLbioanalyticSYSTEMS
Hans-Werner Liebsch
Sales Office Scient. Instruments
Weidenstrasse 11
D - 72119 ENTRINGEN
Germany
Tel.: 0049 (0) 7073 916796
Fax : 0049 (0) 7073 916798
e-mail : hwl.bioanalytic-at-bluewin.de
http://www.hwlbioanalytic.com






From: tracy gales :      tl_gales-at-fccc.edu
Date: Mon, 28 Feb 2000 09:24:29 -0500
Subject: Virology

Contents Retrieved from Microscopy Listserver Archives
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Ronnie,

We use many of the techniques and the atlas from the following excellent
book.
"EM in Diagnostic Virology" by Doan and Anderson,1987, Cambridge
University Press,
ISBN 0 521 24311 4.

--
Tracy Gales
Fox Chase Cancer Center
Electron Microscopy Facility
7701 Burholme Ave.
Philadelphia PA 19111
Tel. 215-728-2484
Fax 215-728-2412






From: Louis M Kerr :      lkerr-at-mbl.edu
Date: Mon, 28 Feb 2000 11:12:56 -0500
Subject: TEM Diffraction plate reader

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A friend recently retired and gave me a TEM diffraction plate reader. This
was made by Ernest F. Fullam, Inc. in the early 1970's. It is still in the
wooden shipping crate, is in unused condition and looks complete except
there is no manual. It is a basic no-frills manual version. It has a high
quality Starrett vernier scale and could be used for measurement of any
electron micrograph, or other type of transparency, or it may be useful for
teaching the history and theory of diffraction pattern measurement.

I would be willing to send it to someone willing to pay the shipping costs.

Thanks,
Louie

Louie Kerr
Research and Education Support Coordinator
Marine Biological Laboratory
7 MBL Street
Woods Hole, MA 02543
508-289-7273
508-540-6902 (FAX)
508-292-0289 (Cell phone)

VISIT OUR WEB SITE:
http://www.mbl.edu






From: Guoda Lian :      glian0-at-engr.uky.edu
Date: Mon, 28 Feb 2000 12:49:27 -0500 (EST)
Subject: Position Available (fwd)

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RESEARCH ASSOCIATE

Electron Microscopy Facility

University of Kentucky


The University of Kentucky invites applications for a Research Associate
in the area of electron microscopy. The Electron Microscopy Facility
houses two SEMs, two TEMs one AFM and extensive sample preparation
equipment that are used by undergraduate and graduate students in
support of research and education. The primary responsibility of the
Research Associate will be to oversee the JEOL 2010F Field Emission TEM
and auxiliary equipment, including an Oxford energy dispersive x-ray
spectrometer (EDS), Gatan electron energy loss imaging filter and
acquisition hardware and software. S/he will be responsible for
coordinating and administering maintenance, access, training and use of
the equipment by internal and external users. S/he will also be
encouraged to develop sponsored research programs and collaborate with
university faculty members. The qualified candidate will have a Ph.D.
and practical experience in analytical electron microscopy and
demonstrate good communication skills. The salary will be commensurate
with qualifications and experience. The University of Kentucky offers
comprehensive insurance and benefits packages and is an equal
opportunity employer. Please send applications to:


Professor Elizabeth Dickey

Director, Electron Microscopy Facility

University of Kentucky

A254 ASTeCC Bldg.

Lexington, KY 40506-0286



***********

Elizabeth C. Dickey

Assistant Professor

Department of Chemical and Materials Engineering

Director, Electron Microscopy Facility

University of Kentucky

177 Anderson Hall

Lexington, KY 40506-0046

ph: 606.257.2300 ext.288

FAX: 606.323.1929

http://www.engr.uky.edu/CME/faculty/dickey/







From: Sara Miller :      saram-at-duke.edu
Date: Mon, 28 Feb 2000 12:38:41 -0500 (EST)
Subject: Re: Need help on EM negative staining

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Ronnie,

We perform about 1000 exams/yr for viruses using EM. Below are some refs
(check out especially 2, 5, 9, 11, 14). Also, at the bottom is a
synopsis I give to our medical students; maybe it will be useful.

As for enteric viruses, EM is the best way to look for them. Many of
them do not grow in tissue culture, and those that can be coaxed, do so
under special conditions that are not employed in the routine culture
lab. Biochemical identification (e.g., immunological kits, PCR, Western
Blots) require a specific reagent to recognize the virus and if you
don't use the right reagent, you will miss viruses (e.g., if you run a
test for rotavirus, you will miss adenovirus, etc.) Furthermore, there
aren't biochemical reagents for all viruses. With EM, you can
see a wide variety of viruses by doing a negative stain of an aqueous
extract of stool.

Good luck,
Sara

REFERENCES ON VIRUS IDENTIFICATION BY ELECTRON MICROSCOPY

1. Cegielski JP, Msengi AE, Miller SE. 1994. Enteric viruses
associated with HIV infection in Tanzanian children with chronic
diarrhea. Ped AIDS HIV Inf 5:296-299.

2. Doane FW, Anderson N. 1986. Electron Microscopy in Diagnostic
Virology: A Practical Guide and Atlas, Cambridge University Press, New York.

3. Lew JF, Glass, RI, Petric M, LeBaron CW, Hammond GW, Miller SE,
Robinson C, Boutilier J, Riepenhoff-Talty M, Payne CM, Franklin R, Oshiro
LS, and Jaqua M-J. 1990. Six year retrospective surveillance of
gastroenteritis viruses identified at ten electron microscopy centers in
the United States and Canada Pediatr Infect Dis J 9:709-714.

4. Mclean DM, Wong KK. 1984. Same Day Diagnosis of Human Virus
Infections, CRC Press, Boca Raton, FL.

5. Miller SE. 1986. Detection and identification of viruses by
electron microscopy. J Electron Microsc Tech 4:265-301.

5. Miller SE, Howell DN. 1988. Viral infections in the acquired
immunodeficiency syndrome. J Electron Microsc Tech 8:41-78.

7. Miller SE. 1988. Diagnostic virology by electron microscopy.
ASM News 54:475-481.

8. Miller SE. 1989. Electron microscopy in rapid viral diagnosis.
EMSA Bull 19:53-59.

9. Hayat MA, Miller SE. 1990. Negative Staining: Applications and
Methods. McGraw-Hill, New York.

10. Miller SE. 1991. Evaluation of electron microscopic information
available from clinical samples. In de la Maza LM, Peterson EM (eds.),
Medical Virology, Vol 10. Irvine, CA. pp 21-54.

11. Miller SE. 1995. Diagnosis of viral infection by electron
microscopy. In Lennette EH, Lennette DA, Lennette ET (eds), Diagnostic
Procedures for Viral, Rickettsial and Chlamydial Infections, American
Public Health Assoc, Washington, DC. pp 35-76.

12. Oshiro LS, Miller SE. 1992. Application of electron microscopy
to the diagnosis of viral infections. In Lennette EH (ed), Laboratory
Diagnosis of Viral Infections. Marcel Dekker, New York. pp 45-68.

13. Palmer E, Martin ML. 1982. An Atlas of Mammalian Viruses, CRC
Press, Boca Raton, FL.

14. Palmer E, Martin ML. 1988. Electron Microscopy in Viral
Diagnosis, CRC Press, Boca Raton, FL.

15. Murphy FA, Fauquet CM, Bishop DHL, Ghabrial SA, Jarvis AW, GP
Martelli, Mayo MA, Summers MD. 1995. Virus Taxonomy. Sixth Report of the
International Committee on Taxonomy of Viruses. Springer-Verlag, New York.

2, 5, 9, 11-14 = Methods and micrographs
2, 14 = Excellent atlases
7, 8 = Brief synopses of technique and basis of virus ID
15 = Good general reference, but don't memorize!! Lists all viruses, not
just human.



TWO WAYS TO LOOK AT VIRUSES BY EM:
Negative staining
Used with liquid samples
Shows whole virus (size, surface structure important)
Thin sectioning
Used with cells and tissues
Shows a slice of an infected cell
Shows relationship of viruses to cell organelles (internal
structure and cell organelle association important)
Location within the cell is a clue to nucleic acid type
DNA viruses are usually seen in the nucleus (some
exceptions).
RNA viruses are usually seen in the cytoplasm
(some exceptions).
Exceptions:
Naked viruses get out of the cell by lysis; if
the cell is very sick, there may be virions
everywhere, e.g., adenovirus (DNA).
Poxviruses are DNA viruses, but are constructed
in the cytoplasm.
Hepatitis B core particles (DNA) may be in the
nucleus and cytoplasm. Dane particles
(complete viruses) are only cytoplasmic.
Some paramyxovirus (RNA) nucleocapsids, not the
complete virion, can be seen in the nucleus
(e.g., measlesvirus).
Enveloped viruses are associated with cell membranes; the
type of cell membrane (plasma membrane,
nuclear membrane, endoplasmic reticulum, vacuoles) can
be a clue to virus identification.

SUMMARY OF VIRUS TYPES (based on morphology):
Naked viruses
All human naked viruses are icosahedral (spherical).
There are 3 size ranges from 20-35 nm, 45-55 nm, 70-90 nm. Examples:
Small: parvovirus, enterovirus, rhinovirus
Medium: polyomavirus, papillomavirus
Large: rotavirus/reovirus, adenovirus
Enveloped viruses
All have a lipoprotein membrane on the outside of the
nucleocapsid. Most derive this layer by budding through
cell membranes; this pliable layer makes the virion
pleomorphic. Poxviruses synthesize their shell de novo;
sometimes they pick up an extra covering by budding. They are
not pleomorphic, but different varieties may be more ovoid,
others, brick-shaped.
Most range in size from 40-400 nm. (Filoviruses are 80 nm by up
to 1400 nm.)
The envelope has proteins on the outside that may look like
spikes or fuzz (e.g., influenza virus, coronavirus).
Sometimes the spikes are too short to be distinguished by EM,
making the virion appear smooth (e.g., herpesviruses,
rubella virus).
The nucleocapsid shape can be
a. icosahedral, like the naked viruses; however, some
are morphologically nondescript as seen by EM;
b. filamentous, like a "slinky."
c. complex (poxvirus cores look like dumbbells with
lateral bodies)
d. morphologically nondescript (just dense without any
particular shape).
Enveloped DNA viruses (except pox) construct their nucleocapsids
in the nucleus.
Enveloped RNA viruses construct their nucleocapsids in the
cytoplasm; (measles nucleocapsids can sometimes be seen
in the nucleus).



On Sat, 26 Feb 2000, Ronald R. Matias wrote:

} Date: Sat, 26 Feb 2000 08:19:49 +0800
} From: Ronald R. Matias {rrm-at-skyinet.net}
} To: Microscopy-at-sparc5.microscopy.com
} Subject: Need help on EM negative staining
}
} ------------------------------------------------------------------------
} 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 and Help!
}
} We are a hospital based research lab. starting to use the electron
} microscope to identify or confirm pathogens that cause gastrointestinal
} disturbances. We would like to know if someone out there could recommend
} appropriate specific EM protocols (negative staining, processing of the
} gastrointestinal contents, etc....) to visualize these pathogens (ranging
} from viruses to bacteria and protozoans). We have a Jeol 1010 EM and perform
} the routine EM preparations designed to examine tissues.
}
} In the interest of bring quality healthcare to our people, we will be very
} much obliged,
}
} ronnie matias
}
}

Sara E. Miller, Ph. D.
P. O. Box 3020
Duke University Medical Center
Durham, NC 27710
Ph: 919 684-3452
FAX: 919 684-8735





From: Sara Miller :      saram-at-duke.edu
Date: Mon, 28 Feb 2000 12:38:41 -0500 (EST)
Subject: Re: Need help on EM negative staining

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html




---------- Forwarded message ----------


Ronnie,

We perform about 1000 exams/yr for viruses using EM. Below are some refs
(check out especially 2, 5, 9, 11, 14). Also, at the bottom is a
synopsis I give to our medical students; maybe it will be useful.

As for enteric viruses, EM is the best way to look for them. Many of
them do not grow in tissue culture, and those that can be coaxed, do so
under special conditions that are not employed in the routine culture
lab. Biochemical identification (e.g., immunological kits, PCR, Western
Blots) require a specific reagent to recognize the virus and if you
don't use the right reagent, you will miss viruses (e.g., if you run a
test for rotavirus, you will miss adenovirus, etc.) Furthermore, there
aren't biochemical reagents for all viruses. With EM, you can
see a wide variety of viruses by doing a negative stain of an aqueous
extract of stool.

Good luck,
Sara

REFERENCES ON VIRUS IDENTIFICATION BY ELECTRON MICROSCOPY

1. Cegielski JP, Msengi AE, Miller SE. 1994. Enteric viruses
associated with HIV infection in Tanzanian children with chronic
diarrhea. Ped AIDS HIV Inf 5:296-299.

2. Doane FW, Anderson N. 1986. Electron Microscopy in Diagnostic
Virology: A Practical Guide and Atlas, Cambridge University Press, New York.

3. Lew JF, Glass, RI, Petric M, LeBaron CW, Hammond GW, Miller SE,
Robinson C, Boutilier J, Riepenhoff-Talty M, Payne CM, Franklin R, Oshiro
LS, and Jaqua M-J. 1990. Six year retrospective surveillance of
gastroenteritis viruses identified at ten electron microscopy centers in
the United States and Canada Pediatr Infect Dis J 9:709-714.

4. Mclean DM, Wong KK. 1984. Same Day Diagnosis of Human Virus
Infections, CRC Press, Boca Raton, FL.

5. Miller SE. 1986. Detection and identification of viruses by
electron microscopy. J Electron Microsc Tech 4:265-301.

5. Miller SE, Howell DN. 1988. Viral infections in the acquired
immunodeficiency syndrome. J Electron Microsc Tech 8:41-78.

7. Miller SE. 1988. Diagnostic virology by electron microscopy.
ASM News 54:475-481.

8. Miller SE. 1989. Electron microscopy in rapid viral diagnosis.
EMSA Bull 19:53-59.

9. Hayat MA, Miller SE. 1990. Negative Staining: Applications and
Methods. McGraw-Hill, New York.

10. Miller SE. 1991. Evaluation of electron microscopic information
available from clinical samples. In de la Maza LM, Peterson EM (eds.),
Medical Virology, Vol 10. Irvine, CA. pp 21-54.

11. Miller SE. 1995. Diagnosis of viral infection by electron
microscopy. In Lennette EH, Lennette DA, Lennette ET (eds), Diagnostic
Procedures for Viral, Rickettsial and Chlamydial Infections, American
Public Health Assoc, Washington, DC. pp 35-76.

12. Oshiro LS, Miller SE. 1992. Application of electron microscopy
to the diagnosis of viral infections. In Lennette EH (ed), Laboratory
Diagnosis of Viral Infections. Marcel Dekker, New York. pp 45-68.

13. Palmer E, Martin ML. 1982. An Atlas of Mammalian Viruses, CRC
Press, Boca Raton, FL.

14. Palmer E, Martin ML. 1988. Electron Microscopy in Viral
Diagnosis, CRC Press, Boca Raton, FL.

15. Murphy FA, Fauquet CM, Bishop DHL, Ghabrial SA, Jarvis AW, GP
Martelli, Mayo MA, Summers MD. 1995. Virus Taxonomy. Sixth Report of the
International Committee on Taxonomy of Viruses. Springer-Verlag, New York.

2, 5, 9, 11-14 = Methods and micrographs
2, 14 = Excellent atlases
7, 8 = Brief synopses of technique and basis of virus ID
15 = Good general reference, but don't memorize!! Lists all viruses, not
just human.



TWO WAYS TO LOOK AT VIRUSES BY EM:
Negative staining
Used with liquid samples
Shows whole virus (size, surface structure important)
Thin sectioning
Used with cells and tissues
Shows a slice of an infected cell
Shows relationship of viruses to cell organelles (internal
structure and cell organelle association important)
Location within the cell is a clue to nucleic acid type
DNA viruses are usually seen in the nucleus (some
exceptions).
RNA viruses are usually seen in the cytoplasm
(some exceptions).
Exceptions:
Naked viruses get out of the cell by lysis; if
the cell is very sick, there may be virions
everywhere, e.g., adenovirus (DNA).
Poxviruses are DNA viruses, but are constructed
in the cytoplasm.
Hepatitis B core particles (DNA) may be in the
nucleus and cytoplasm. Dane particles
(complete viruses) are only cytoplasmic.
Some paramyxovirus (RNA) nucleocapsids, not the
complete virion, can be seen in the nucleus
(e.g., measlesvirus).
Enveloped viruses are associated with cell membranes; the
type of cell membrane (plasma membrane,
nuclear membrane, endoplasmic reticulum, vacuoles) can
be a clue to virus identification.

SUMMARY OF VIRUS TYPES (based on morphology):
Naked viruses
All human naked viruses are icosahedral (spherical).
There are 3 size ranges from 20-35 nm, 45-55 nm, 70-90 nm. Examples:
Small: parvovirus, enterovirus, rhinovirus
Medium: polyomavirus, papillomavirus
Large: rotavirus/reovirus, adenovirus
Enveloped viruses
All have a lipoprotein membrane on the outside of the
nucleocapsid. Most derive this layer by budding through
cell membranes; this pliable layer makes the virion
pleomorphic. Poxviruses synthesize their shell de novo;
sometimes they pick up an extra covering by budding. They are
not pleomorphic, but different varieties may be more ovoid,
others, brick-shaped.
Most range in size from 40-400 nm. (Filoviruses are 80 nm by up
to 1400 nm.)
The envelope has proteins on the outside that may look like
spikes or fuzz (e.g., influenza virus, coronavirus).
Sometimes the spikes are too short to be distinguished by EM,
making the virion appear smooth (e.g., herpesviruses,
rubella virus).
The nucleocapsid shape can be
a. icosahedral, like the naked viruses; however, some
are morphologically nondescript as seen by EM;
b. filamentous, like a "slinky."
c. complex (poxvirus cores look like dumbbells with
lateral bodies)
d. morphologically nondescript (just dense without any
particular shape).
Enveloped DNA viruses (except pox) construct their nucleocapsids
in the nucleus.
Enveloped RNA viruses construct their nucleocapsids in the
cytoplasm; (measles nucleocapsids can sometimes be seen
in the nucleus).



On Sat, 26 Feb 2000, Ronald R. Matias wrote:

} Date: Sat, 26 Feb 2000 08:19:49 +0800
} From: Ronald R. Matias {rrm-at-skyinet.net}
} To: Microscopy-at-sparc5.microscopy.com
} Subject: Need help on EM negative staining
}
} ------------------------------------------------------------------------
} 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 and Help!
}
} We are a hospital based research lab. starting to use the electron
} microscope to identify or confirm pathogens that cause gastrointestinal
} disturbances. We would like to know if someone out there could recommend
} appropriate specific EM protocols (negative staining, processing of the
} gastrointestinal contents, etc....) to visualize these pathogens (ranging
} from viruses to bacteria and protozoans). We have a Jeol 1010 EM and perform
} the routine EM preparations designed to examine tissues.
}
} In the interest of bring quality healthcare to our people, we will be very
} much obliged,
}
} ronnie matias
}
}

Sara E. Miller, Ph. D.
P. O. Box 3020
Duke University Medical Center
Durham, NC 27710
Ph: 919 684-3452
FAX: 919 684-8735






From: Sara Miller :      saram-at-duke.edu
Date: Mon, 28 Feb 2000 12:38:41 -0500 (EST)
Subject: Re: Need help on EM negative staining

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Ronnie,

We perform about 1000 exams/yr for viruses using EM. Below are some refs
(check out especially 2, 5, 9, 11, 14). Also, at the bottom is a
synopsis I give to our medical students; maybe it will be useful.

As for enteric viruses, EM is the best way to look for them. Many of
them do not grow in tissue culture, and those that can be coaxed, do so
under special conditions that are not employed in the routine culture
lab. Biochemical identification (e.g., immunological kits, PCR, Western
Blots) require a specific reagent to recognize the virus and if you
don't use the right reagent, you will miss viruses (e.g., if you run a
test for rotavirus, you will miss adenovirus, etc.) Furthermore, there
aren't biochemical reagents for all viruses. With EM, you can
see a wide variety of viruses by doing a negative stain of an aqueous
extract of stool.

Good luck,
Sara

REFERENCES ON VIRUS IDENTIFICATION BY ELECTRON MICROSCOPY

1. Cegielski JP, Msengi AE, Miller SE. 1994. Enteric viruses
associated with HIV infection in Tanzanian children with chronic
diarrhea. Ped AIDS HIV Inf 5:296-299.

2. Doane FW, Anderson N. 1986. Electron Microscopy in Diagnostic
Virology: A Practical Guide and Atlas, Cambridge University Press, New York.

3. Lew JF, Glass, RI, Petric M, LeBaron CW, Hammond GW, Miller SE,
Robinson C, Boutilier J, Riepenhoff-Talty M, Payne CM, Franklin R, Oshiro
LS, and Jaqua M-J. 1990. Six year retrospective surveillance of
gastroenteritis viruses identified at ten electron microscopy centers in
the United States and Canada Pediatr Infect Dis J 9:709-714.

4. Mclean DM, Wong KK. 1984. Same Day Diagnosis of Human Virus
Infections, CRC Press, Boca Raton, FL.

5. Miller SE. 1986. Detection and identification of viruses by
electron microscopy. J Electron Microsc Tech 4:265-301.

5. Miller SE, Howell DN. 1988. Viral infections in the acquired
immunodeficiency syndrome. J Electron Microsc Tech 8:41-78.

7. Miller SE. 1988. Diagnostic virology by electron microscopy.
ASM News 54:475-481.

8. Miller SE. 1989. Electron microscopy in rapid viral diagnosis.
EMSA Bull 19:53-59.

9. Hayat MA, Miller SE. 1990. Negative Staining: Applications and
Methods. McGraw-Hill, New York.

10. Miller SE. 1991. Evaluation of electron microscopic information
available from clinical samples. In de la Maza LM, Peterson EM (eds.),
Medical Virology, Vol 10. Irvine, CA. pp 21-54.

11. Miller SE. 1995. Diagnosis of viral infection by electron
microscopy. In Lennette EH, Lennette DA, Lennette ET (eds), Diagnostic
Procedures for Viral, Rickettsial and Chlamydial Infections, American
Public Health Assoc, Washington, DC. pp 35-76.

12. Oshiro LS, Miller SE. 1992. Application of electron microscopy
to the diagnosis of viral infections. In Lennette EH (ed), Laboratory
Diagnosis of Viral Infections. Marcel Dekker, New York. pp 45-68.

13. Palmer E, Martin ML. 1982. An Atlas of Mammalian Viruses, CRC
Press, Boca Raton, FL.

14. Palmer E, Martin ML. 1988. Electron Microscopy in Viral
Diagnosis, CRC Press, Boca Raton, FL.

15. Murphy FA, Fauquet CM, Bishop DHL, Ghabrial SA, Jarvis AW, GP
Martelli, Mayo MA, Summers MD. 1995. Virus Taxonomy. Sixth Report of the
International Committee on Taxonomy of Viruses. Springer-Verlag, New York.

2, 5, 9, 11-14 = Methods and micrographs
2, 14 = Excellent atlases
7, 8 = Brief synopses of technique and basis of virus ID
15 = Good general reference, but don't memorize!! Lists all viruses, not
just human.



TWO WAYS TO LOOK AT VIRUSES BY EM:
Negative staining
Used with liquid samples
Shows whole virus (size, surface structure important)
Thin sectioning
Used with cells and tissues
Shows a slice of an infected cell
Shows relationship of viruses to cell organelles (internal
structure and cell organelle association important)
Location within the cell is a clue to nucleic acid type
DNA viruses are usually seen in the nucleus (some
exceptions).
RNA viruses are usually seen in the cytoplasm
(some exceptions).
Exceptions:
Naked viruses get out of the cell by lysis; if
the cell is very sick, there may be virions
everywhere, e.g., adenovirus (DNA).
Poxviruses are DNA viruses, but are constructed
in the cytoplasm.
Hepatitis B core particles (DNA) may be in the
nucleus and cytoplasm. Dane particles
(complete viruses) are only cytoplasmic.
Some paramyxovirus (RNA) nucleocapsids, not the
complete virion, can be seen in the nucleus
(e.g., measlesvirus).
Enveloped viruses are associated with cell membranes; the
type of cell membrane (plasma membrane,
nuclear membrane, endoplasmic reticulum, vacuoles) can
be a clue to virus identification.

SUMMARY OF VIRUS TYPES (based on morphology):
Naked viruses
All human naked viruses are icosahedral (spherical).
There are 3 size ranges from 20-35 nm, 45-55 nm, 70-90 nm. Examples:
Small: parvovirus, enterovirus, rhinovirus
Medium: polyomavirus, papillomavirus
Large: rotavirus/reovirus, adenovirus
Enveloped viruses
All have a lipoprotein membrane on the outside of the
nucleocapsid. Most derive this layer by budding through
cell membranes; this pliable layer makes the virion
pleomorphic. Poxviruses synthesize their shell de novo;
sometimes they pick up an extra covering by budding. They are
not pleomorphic, but different varieties may be more ovoid,
others, brick-shaped.
Most range in size from 40-400 nm. (Filoviruses are 80 nm by up
to 1400 nm.)
The envelope has proteins on the outside that may look like
spikes or fuzz (e.g., influenza virus, coronavirus).
Sometimes the spikes are too short to be distinguished by EM,
making the virion appear smooth (e.g., herpesviruses,
rubella virus).
The nucleocapsid shape can be
a. icosahedral, like the naked viruses; however, some
are morphologically nondescript as seen by EM;
b. filamentous, like a "slinky."
c. complex (poxvirus cores look like dumbbells with
lateral bodies)
d. morphologically nondescript (just dense without any
particular shape).
Enveloped DNA viruses (except pox) construct their nucleocapsids
in the nucleus.
Enveloped RNA viruses construct their nucleocapsids in the
cytoplasm; (measles nucleocapsids can sometimes be seen
in the nucleus).



On Sat, 26 Feb 2000, Ronald R. Matias wrote:

} Date: Sat, 26 Feb 2000 08:19:49 +0800
} From: Ronald R. Matias {rrm-at-skyinet.net}
} To: Microscopy-at-sparc5.microscopy.com
} Subject: Need help on EM negative staining
}
} ------------------------------------------------------------------------
} 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 and Help!
}
} We are a hospital based research lab. starting to use the electron
} microscope to identify or confirm pathogens that cause gastrointestinal
} disturbances. We would like to know if someone out there could recommend
} appropriate specific EM protocols (negative staining, processing of the
} gastrointestinal contents, etc....) to visualize these pathogens (ranging
} from viruses to bacteria and protozoans). We have a Jeol 1010 EM and perform
} the routine EM preparations designed to examine tissues.
}
} In the interest of bring quality healthcare to our people, we will be very
} much obliged,
}
} ronnie matias
}
}

Sara E. Miller, Ph. D.
P. O. Box 3020
Duke University Medical Center
Durham, NC 27710
Ph: 919 684-3452
FAX: 919 684-8735






From: Guoda Lian :      glian0-at-engr.uky.edu
Date: Mon, 28 Feb 2000 12:49:27 -0500 (EST)
Subject: Position Available (fwd)

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html




---------- Forwarded message ----------




---------- Forwarded message ----------





RESEARCH ASSOCIATE

Electron Microscopy Facility

University of Kentucky


The University of Kentucky invites applications for a Research Associate
in the area of electron microscopy. The Electron Microscopy Facility
houses two SEMs, two TEMs one AFM and extensive sample preparation
equipment that are used by undergraduate and graduate students in
support of research and education. The primary responsibility of the
Research Associate will be to oversee the JEOL 2010F Field Emission TEM
and auxiliary equipment, including an Oxford energy dispersive x-ray
spectrometer (EDS), Gatan electron energy loss imaging filter and
acquisition hardware and software. S/he will be responsible for
coordinating and administering maintenance, access, training and use of
the equipment by internal and external users. S/he will also be
encouraged to develop sponsored research programs and collaborate with
university faculty members. The qualified candidate will have a Ph.D.
and practical experience in analytical electron microscopy and
demonstrate good communication skills. The salary will be commensurate
with qualifications and experience. The University of Kentucky offers
comprehensive insurance and benefits packages and is an equal
opportunity employer. Please send applications to:


Professor Elizabeth Dickey

Director, Electron Microscopy Facility

University of Kentucky

A254 ASTeCC Bldg.

Lexington, KY 40506-0286



***********

Elizabeth C. Dickey

Assistant Professor

Department of Chemical and Materials Engineering

Director, Electron Microscopy Facility

University of Kentucky

177 Anderson Hall

Lexington, KY 40506-0046

ph: 606.257.2300 ext.288

FAX: 606.323.1929

http://www.engr.uky.edu/CME/faculty/dickey/








From: Guoda Lian :      glian0-at-engr.uky.edu
Date: Mon, 28 Feb 2000 12:49:27 -0500 (EST)
Subject: Position Available (fwd)

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html





RESEARCH ASSOCIATE

Electron Microscopy Facility

University of Kentucky


The University of Kentucky invites applications for a Research Associate
in the area of electron microscopy. The Electron Microscopy Facility
houses two SEMs, two TEMs one AFM and extensive sample preparation
equipment that are used by undergraduate and graduate students in
support of research and education. The primary responsibility of the
Research Associate will be to oversee the JEOL 2010F Field Emission TEM
and auxiliary equipment, including an Oxford energy dispersive x-ray
spectrometer (EDS), Gatan electron energy loss imaging filter and
acquisition hardware and software. S/he will be responsible for
coordinating and administering maintenance, access, training and use of
the equipment by internal and external users. S/he will also be
encouraged to develop sponsored research programs and collaborate with
university faculty members. The qualified candidate will have a Ph.D.
and practical experience in analytical electron microscopy and
demonstrate good communication skills. The salary will be commensurate
with qualifications and experience. The University of Kentucky offers
comprehensive insurance and benefits packages and is an equal
opportunity employer. Please send applications to:


Professor Elizabeth Dickey

Director, Electron Microscopy Facility

University of Kentucky

A254 ASTeCC Bldg.

Lexington, KY 40506-0286



***********

Elizabeth C. Dickey

Assistant Professor

Department of Chemical and Materials Engineering

Director, Electron Microscopy Facility

University of Kentucky

177 Anderson Hall

Lexington, KY 40506-0046

ph: 606.257.2300 ext.288

FAX: 606.323.1929

http://www.engr.uky.edu/CME/faculty/dickey/








From: Michael Bode :      mb-at-soft-imaging.com
Date: Mon, 28 Feb 2000 11:31:04 -0700
Subject: FW: Image analysis

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Supanee,

I don't know if you have received an answer since you posted this
message, but if you'd like, we could have a look at your images and try
to help you. What we need is of course the image(s) and a description of
what you actually want to measure. Please be as specific as you can.
Please send the material to my email address below.

Thanks.

Michael Bode, Ph.D.
Soft Imaging System Corp.
1675 Carr St., #105N
Lakewood, CO 80215
===================================
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: Exchange Administrator
Sent: Wednesday, February 23, 2000 8:35 PM
To: Michael Bode


Dear all,
I am doing the image analysis of SEM micrographs. The
micrographs contain both dense (light) and air cell (dark) areas. But
two
areas are not discrete. I'm having a difficulty of quantifying those
separated fractions. I wonder if anyone has any ideas of how to
solve the problem.
Looking forward for your suggestions.
Regards,
Supanee





From: George Lawton :      George.Lawton-at-email.swmed.edu
Date: Mon, 28 Feb 2000 12:48:57 -0600
Subject: TEM - Embedding Problems

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Occassionally someone sends a question to the listserver and there is no response. It doesn't happen often so I thought I would send this question again.


Several weeks ago, I was given some inner membranes from E. coli to be run up to Epon and sectioned for TEM. I treated the sample like tissue culture and pelleted the membranes. Fixed in
3% glut in PBS and post-fixed in 1% OsO4. Routine dehydration in alcohols. Embedded in
EMbed 812 with BDMA in Beem Capsules. Left in oven 48 hours. When I tried to section, the tissue was too soft and appeared to not be infiltrated well. Put back into the oven for 2-3 more days. No difference. Thinking I had a neural meltdown while processing the samples, I tried again on fresh samples, paying particular attention to details. I also used new bottles of embedding media. Same results.
The part of the block without the tissue is its usual hardness. The part of the block with the tissue
(the tip of the beem capsule) is soft and tacky.

Has anyone experienced anything like this? Any suggestions on how I can get the sample into epon?

Thanks in advance for any ideas and/or suggestions.

George Lawton
Chief Electron Microscopist
Molecular and Cellular Imaging Facility
UT Southwestern Medical Center at Dallas
5323 Harry Hines Blvd.
Dallas, Tx 75390-9039
Phone: 214-648-7291
eMail: George.Lawton-at-email.swmed.edu




From: Greg Erdos :      gwe-at-biotech.ufl.edu
Date: Mon, 28 Feb 2000 15:08:21 -0500
Subject: Re: TEM - Embedding Problems

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


If you were working with a pellet in the bottom of a tube or BEEm cap, it
is possible that you were not getting good exchange of reagents at the tip
of the tube during dehydration and embedding.
Or is you were centifuging in the final resin mixture before
putting into the oven it is possible that your sample was packed so tightly
that there was not enough resin in it to polymerize properly.


At 12:48 PM 02/28/2000 -0600, you wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

Gregory W. Erdos, Ph.D. Ph. 352-392-1295
Assistant Director, Biotechnology Program
PO Box 110580 Fax:
352-846-0251
University of Florida
Gainesville, FL 32611






From: Margie Bryant :      mbryant-at-com1.med.usf.edu
Date: Feb.28, 2000
Subject: Leitz Ultrapak incident-Light illuminator

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html



Hello,

We have an old Leitz Orthoplan microscope and are
looking for the Ultrapak incident light illuminator (housing with fixed
ring mirror and interchangeable UO objectives of 4X to 60X primary
magnification). These components have not been produced in many years.
If anyone has this equipment, we would be pleased to pay for it and for
the shipping. Thank you very much!

Sincerely,

Margie Bryant

From: Margie
Bryant {mbryant-at-com1.med.usf.edu





From: przybylo :      przybylowicz-at-nac.ac.za
Date: Mon, 28 Feb 2000 23:08:46 +0200
Subject: As in biological tissues

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Dear Doug,

We have some experience in using micro-PIXE (proton induced X-ray emission)
and the so-called nuclear microprobe in toxicological research. It is more
sensitive than EDS and WDS, is very easy to quantify (standardless
technique). In principle you can have detection limits of the order of 1
ppm in point analysis mode. Specimens can be of any thickness and their
actual thickness is assessed by the simultaneous use of the BS technique
(proton backscatering spectrometry). What in my opinion makes it really
attractive for this purpose is its very good scanning capability. Scan sizes
are from ca. 2.5 mm x 2.5 mm down to the beam resolution, which is of the
order of few micrometer (can be down to 1 micrometer or even less, but in
most of our uses few micrometers is good enough).
The sample preparation is similar as for EDS, using cryotechniques to avoid
elemental losses and redistribution.
We have so far done studies on Ni, Mo, Cd. The detection limits for As are
better than for Mo and Cd.
Of course, this technique cannot indicate the state of oxidation, you can
really treat it as a "more sensitive EDS".

If you want more information, I can refer you to our earlier work and
perhaps you might be interested in contacting me.

Best regards


xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
Dr Wojciech J. Przybylowicz
Materials Research Group
National Accelerator Centre
PO Box 72
Faure 7131 South Africa
E-mail: przybylowicz-at-nac.ac.za
Fax: +27-21-8433543
Phone: +27-21-8433820
http://www.nac.ac.za/IBA/
http://www.nac.ac.za/materials/
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx





From: rlvaughn-at-unmc.edu
Date: Mon, 28 Feb 2000 15:15:58 -0600
Subject: network printing management

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


I seem to recall a discussion of software that was available to keep track
of printing jobs submitted to a network printer for billing purposes? An
argument here for not installing a networked dyesub printer for digital
images is who is going to be responsible for watching who's using it and
how many prints they're making. We have had some problems like this with
the Epson 850 which is on the network and somebody has ran through two
color cartridges in a short time. Thanks for any advice.

Rick Vaughn
rlvaughn-at-unmc.edu





From: John J. Bozzola :      bozzola-at-siu.edu
Date: Mon, 28 Feb 2000 17:27:59 -0600
Subject: Image Analysis of Spheres

Contents Retrieved from Microscopy Listserver Archives
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We are looking for a program (preferably Macintosh-based) that will
automatically measure the diameter of spheres that have been imaged
in a TEM.

Typically, the spheres measure about 3-5 mm on the TEM negative and
often are touching or slightly overlapped. I know that NIH Image
should do this, but I have not been able to get reproduceable data
using this program.

Someone told me about a program called "Bolero", written by some
Spanish university researchers but it is an $8-10K program. Too rich
for our tastes.

Any suggestions?

We do appreciate your suggestions.

John

####################################################################
John J. Bozzola, Ph.D., Director
Micro-Imaging and Analysis Center
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
####################################################################




From: William McManus :      billemac-at-biology.usu.edu
Date: Mon, 28 Feb 2000 16:36:18 -0700
Subject: bio: peptidoglycan

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Does anyone know of a way to label for peptidoglycan at the light or
electron microscope level?

William McManus
Supervisor
Electron Microscopy Facility
Department of Biology
Utah State University
Logan UT 84322-5305

billEMac-at-cc.usu.edu
435-797-1920





From: Sara Miller :      saram-at-duke.edu
Date: Mon, 28 Feb 2000 18:42:51 -0500 (EST)
Subject: Re: TEM - Embedding Problems

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I agree with Greg: sounds like an infiltration problem. Your best bet
is to encase the bugs in agar. One way is to fix briefly (2-5 min) in glut,
then pellet and let sit for a while (1 hr). This glues them together.

Don't resuspend them in the glut or it will fix the walls of the bugs so
that they won't stick together.

Cut off the bottom of the
tube if plastic or scrape out the cells if tube is glass, onto Parafilm
and drain with wedge of filter paper. Divide into 1 cubic mm (or
smaller) portions and cover with molter agar. Cut away any excess.
Wash. Osmicate and embed as you would a piece of tissue.

Other folks have suspended bugs in molter agar and pelleted them through
the agar. You have to do this before the agar cools. Then cut off the
end with the cells.

Don't fix the agar/cells in glut; it will cross-link the agar so that the
subsequent solutions can't get in. The glut left in the bugs after
they're fixed and before they're encased is OK and will wash out.

Hope this helps.

Sara


Sara E. Miller, Ph. D.
P. O. Box 3020
Duke University Medical Center
Durham, NC 27710
Ph: 919 684-3452
FAX: 919 684-8735





From: DrJohnRuss-at-aol.com
Date: Mon, 28 Feb 2000 18:49:03 EST
Subject: Re: Image Analysis of Spheres

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In a message dated 2/28/00 11:44:37 PM, bozzola-at-siu.edu writes:

} Typically, the spheres measure about 3-5 mm on the TEM negative and
} often are touching or slightly overlapped. I know that NIH Image
} should do this, but I have not been able to get reproduceable data
} using this program.

If none of the spheres have been cut off in the section so that you have
stereological corrections to make for the missing parts, and you aren't
having problems with thresholding, etc., then I don't see what your problem
is. Use a watershed to separate the touching features, and use the maximum
feret's diameter to estimate the sphere diameter, not the area (which may be
reduced due to overaps). Or you can use the maximum values of the euclidean
distance map (the ultimate eroded point) as the sphere radii and not even
bother with the watershed.




From: rachel.harding-at-oxinst.co.uk
Date: Mon, 28 Feb 2000 19:10:48 -0600
Subject: info

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Hello,
Is there a web site where I can find more information about quantitative
techniques in microscopy on SEM's?
In particular the XPP correction procedure?

Many thanks
Rachel


____________________________________________________________
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HP12 3SE, UK Tel: +44 (0)1494 442255 Fax: +44 (0)1494 524129
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and delete it from your records. Internet communications are not secure
and Oxford Instruments is not responsible for their abuse by third
parties nor for any alteration or corruption in transmission.
____________________________________________________________






From: mtl :      mtl-at-njcc.com
Date: Fri, 01 Jan 1999 21:33:41 -0400
Subject: Re: Leitz Ultrapak incident-Light illuminator

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You may want to try Triad Scientific Inc. in Edison, NJ. ,
triadsci-at-webspan.net or (800) 867-6690. When I was there last month to
purchase a vacuum oven, a whole truck lot of optical microscopes and parts
(Leitz, etc.) was being unloaded. Most were still in the original boxes.
J. Roy Nelson
Material Testing Laboratory
Pennington, NJ
mtl-at-njcc.com

Margie Bryant 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
} -----------------------------------------------------------------------.
}
} Date: Feb.28, 2000
}
} Hello,
}
} We have an old Leitz Orthoplan microscope and are
} looking for the Ultrapak incident light illuminator (housing with fixed
} ring mirror and interchangeable UO objectives of 4X to 60X primary
} magnification). These components have not been produced in many years.
} If anyone has this equipment, we would be pleased to pay for it and for
} the shipping. Thank you very much!
}
} Sincerely,
}
} Margie Bryant
}
} From: Margie
} Bryant {mbryant-at-com1.med.usf.edu





From: Marilyn Henderson :      marilyn.henderson-at-adelaide.edu.au
Date: Tue, 29 Feb 2000 13:57:34 +1030
Subject: Re: TEM - Embedding Problems

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Dear George,

I've found the same problem sometimes with Epon/Araldite in Eppendorf
tubes. I think the pellet is so tightly bound to the bottom of the tube
by the time 100% resin infiltration is reached (especially if you
centrifuge along the way) that the resin doesn't have good access to the
cells or material at the 'bottom'. I now lift (ease gently) the pellet
off the bottom of the tube with a toothpick to ensure that the media are
totally surrounding the pellet.
Another choice is to cut away all the gooey resin and hope that the
material at the 'top' of the pellet is well infiltrated and polymerized.

Regards,
Marilyn

George Lawton 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
} -----------------------------------------------------------------------.
}
} Occassionally someone sends a question to the listserver and there is no response. It doesn't happen often so I thought I would send this question again.
}
} Several weeks ago, I was given some inner membranes from E. coli to be run up to Epon and sectioned for TEM. I treated the sample like tissue culture and pelleted the membranes. Fixed in
} 3% glut in PBS and post-fixed in 1% OsO4. Routine dehydration in alcohols. Embedded in
} EMbed 812 with BDMA in Beem Capsules. Left in oven 48 hours. When I tried to section, the tissue was too soft and appeared to not be infiltrated well. Put back into the oven for 2-3 more days. No difference. Thinking I had a neural meltdown while processing the samples, I tried again on fresh samples, paying particular attention to details. I also used new bottles of embedding media. Same results.
} The part of the block without the tissue is its usual hardness. The part of the block with the tissue
} (the tip of the beem capsule) is soft and tacky.
}
} Has anyone experienced anything like this? Any suggestions on how I can get the sample into epon?
}
} Thanks in advance for any ideas and/or suggestions.
}
} George Lawton
} Chief Electron Microscopist
} Molecular and Cellular Imaging Facility
} UT Southwestern Medical Center at Dallas
} 5323 Harry Hines Blvd.
} Dallas, Tx 75390-9039
} Phone: 214-648-7291
} eMail: George.Lawton-at-email.swmed.edu




From: Tina Carvalho :      tina-at-pbrc.hawaii.edu
Date: Mon, 28 Feb 2000 18:37:42 -1000 (HST)
Subject: Re: TEM - Embedding Problems

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Hi-

Two things come to mind. First, if processed in a microfuge tube, loosen
the pellet from the tube and make sure all fluids and epoxy mixtures come
into contact with all surfaces (as already recommended). Second, I always
make it a practice to preheat the BEEM capsules and labels in the
embedding oven all day or overnight before use to drive off any
moisture. This really seems to help in our humid environment!

Good luck and aloha,
Tina

Sunny and 80F but with mosquitoes.

****************************************************************************
* 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*
****************************************************************************





From: Stanley L. Flegler :      flegler-at-pilot.msu.edu
Date: Tue, 29 Feb 2000 08:20:26 -0500
Subject: Re: network printing management

Contents Retrieved from Microscopy Listserver Archives
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Rick:
What you want to do is fairly easy if you are using Windows NT. It will
maintain a log of the user name, number of prints, date, name of print job,
etc. I posted instructions to this group in 1997. These instructions were
published in Microscopy and Microanalysis Vol.3, number 6, p. 569. If you
don't have access to this, let me know and I will send the directions.
Stanley L. Flegler, Acting Director
Center for Advanced Microscopy
Michigan State University




From: mccu5iv2-at-fs2.ee.umist.ac.uk ()
Date: Tue, 29 Feb 2000 07:19:44 -0600
Subject: HREM of semiconductors

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Email: mccu5iv2-at-fs2.ee.umist.ac.uk
Name: Imran

School: Vahora

Question: Currently studying for a project on HREM of semiconductors,
namely GaN; I would like to know where I can obtain some good information
on this subject. My lecturer does not have much insight in this area, I
need to study its structure, and I am currently using the Cerius(2)
software package to simulate images of GaN.

thankyou














---------------------------------------------------------------------------






From: Duncan Waddell :      D.Waddell-at-mailbox.uq.edu.au
Date: Tue, 29 Feb 2000 07:31:01 -0600
Subject: CyberSTEM demonstration

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The Centre for Microscopy & Microanalysis (CMM) at The University of
Queensland will demonstrate its web-based interactive Virtual Electron
Microscopy service this Thursday (March 2, 2000) between 11am and 1pm.
(Queensland Time; GMT +10)

CyberSTEM (WWW accessible Scanning and Transmission Electron Microscopy)
is a service that became officially permanent on the web after Centre
Directors approved funding last week.

URL: http://www.uq.edu.au/nanoworld/online.html

The CMM's videoconference service - begun in 1995 - has evolved into a
webcam-based service that allows unlimited live connections to
microscopy sessions conducted at the Centre. Visitors can interact with
staff during organised sessions via telephone, by chat programs or via
email. (Please note: Live interaction is only available during
specifically organised sessions - direct all other inquiries to the
address/email noted in the signature below.)

Though primarily intended to service paying customers who cannot
physically attend sessions, video streaming from two of their
microscopes (more later they hope) will be regularly accessible by the
general public.

Sincerely,

Duncan

--
************************************************************
Duncan Waddell (BSc)
Senior Scientific Officer
Centre for Microscopy and Microanalysis
The University of Queensland, St. Lucia, Qld, 4072 Australia
Telephone: +61-7-3365-4216
Facsimile: +61-7-3365-4422
WWW: http://www.uq.edu.au/nanoworld/nanohome.html
************************************************************
Any opinion expressed is that of the writer,
and not necessarily that of CMM or of the University.
************************************************************






From: Maria.Fazio-Zanakis-at-aventis.com
Date: Tue, 29 Feb 2000 07:37:17 -0600
Subject: TEM - Embedding Problems

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Dear George,
In all my experience working up bacteria and bacterial fragments, I have
always, and by the way this is the simplest, filtered all suspended cells on
Millipore filters and processed the samples on the filters. This yielded
excellent results. The fragments, cells stayed on the filters and never
detached.
Filter your suspended fragment solution using a 0.2um filter unit system
attached to a vacuum system. The aspirator of a faucet works great. Rinse
in buffer while on the filter system.
Then remove the filter and gently place in a plastic or glass petri dish.
Again, gently drop fixative over the sample enough to cover your filter and
carefully slice it into your appropriate sizes and process from there.
You will have success. If you have further questions, just give me a call.

Blessings,
Maria

Maria Fazio-Zanakis
Bioimaging and Molecular Histology
Aventis Pharmaceuticals
1-908-231-3357
Fax: 1-908-231-3962
Email: Maria.Fazio-Zanakis-at-Aventis.com


-----Original Message-----
} From: George Lawton [mailto:George.Lawton-at-email.swmed.edu]
Sent: Monday, February 28, 2000 1:49 PM
To: microscopy-at-sparc5.microscopy.com


Occassionally someone sends a question to the listserver and there is no
response. It doesn't happen often so I thought I would send this question
again.


Several weeks ago, I was given some inner membranes from E. coli to be run
up to Epon and sectioned for TEM. I treated the sample like tissue culture
and pelleted the membranes. Fixed in
3% glut in PBS and post-fixed in 1% OsO4. Routine dehydration in alcohols.
Embedded in
EMbed 812 with BDMA in Beem Capsules. Left in oven 48 hours. When I tried
to section, the tissue was too soft and appeared to not be infiltrated well.
Put back into the oven for 2-3 more days. No difference. Thinking I had a
neural meltdown while processing the samples, I tried again on fresh
samples, paying particular attention to details. I also used new bottles of
embedding media. Same results.
The part of the block without the tissue is its usual hardness. The part of
the block with the tissue
(the tip of the beem capsule) is soft and tacky.

Has anyone experienced anything like this? Any suggestions on how I can get
the sample into epon?

Thanks in advance for any ideas and/or suggestions.

George Lawton
Chief Electron Microscopist
Molecular and Cellular Imaging Facility
UT Southwestern Medical Center at Dallas
5323 Harry Hines Blvd.
Dallas, Tx 75390-9039
Phone: 214-648-7291
eMail: George.Lawton-at-email.swmed.edu




From: Earl Weltmer :      earlw-at-pacbell.net
Date: Tue, 29 Feb 2000 06:23:04 -0800
Subject: S-520

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Hi all,

I have a customer who wants to purchase a Hitachi S-520 (or equivalent)
SEM with EDS.

If anyone has anything, please contact me off line.

Thank You,

Earl Weltmer
(714) 573-9158







From: Earl Weltmer :      earlw-at-pacbell.net
Date: Tue, 29 Feb 2000 06:23:18 -0800
Subject: S-520

Contents Retrieved from Microscopy Listserver Archives
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Hi all,

I have a customer who wants to purchase a Hitachi S-520 (or equivalent)
SEM with EDS.

If anyone has anything, please contact me off line.

Thank You,

Earl Weltmer
(714) 573-9158







From: Fiona Russell :      fiona.russell-at-gmx.net
Date: Tue, 29 Feb 2000 16:23:06 +0100 (MET)
Subject: Help on: Plant Cell Chromosome Counting

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Can anyone help me?
I am doing research on the plant Achillea millefolium (Yarrow)and I need
to find out the ploidy level of 2 seed sources. I am looking for information
on companies/institutions that will do this work for me and a rough estimate
of the cost.
Many thanks,
Fiona Russell.

--
Sent through Global Message Exchange - http://www.gmx.net





From: Steve Miller :      smiller-at-ventanamed.com
Date: Tue, 29 Feb 2000 13:37:31 -0700
Subject: unsubscribe

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From: HILDEGARD CROWLEY :      hcrowley-at-du.edu
Date: Tue, 29 Feb 2000 15:20:37 -0700 (MST)
Subject: Oversimplification-Sorry

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Hello,

I recently tried to help out Dr. M. Rohde who was embedding in Spurr's
formulation and attempting to locate an antigen.

I said that Spurr's formulation has no place in immunocytochemistry for
all the reasons I listed then. This is an oversimplification (I am always
harassed for time when trying to answer questions completely.) Let me
tell you that I, myself, worked with a graduate student who used Spurr's
medium, etched for one hour in sodium metaperiodate, and then got
wonderful Au label. The reason for this success was that she had an
antigen that was so sturdy and such plentiful supply that it was a
candidate for the prozone effect. Her same exact protocol (actually mine)
when tried by us in our laboratory was a spectacular failure with our
antigen which we soon discovered was scarce and delicate and needed
amplification up to the point where I wondered whether we were actually
increasing the thickness of the section by amplification methods! Sort of
like painting a wall until the room gets smaller!
At any rate - yes, Spurr's can be used for immunocytochemistry. However,
if at first you don't succeed, don't try again. Go to another epoxy, and
then to an acrylic.
I should not make absolute statements as I did. I did not mean to
discourage those who successfully use Spurr's to abandon it. Besides,
compared to what there is to be known about immunocytochemistry, my amount
of knowledge can be dropped into a thimble with room left to stuff an
entire thumb in after it.

I always reserve the right to be wrong!

Hildy Crowley
Sr. Electron Microscopy Specialist
Dep. of Biol. Sciences
University of Denver
Denver, CO





From: Karen Zaruba :      kszaruba-at-MMM.COM
Date: Tue, 29 Feb 2000 16:38:26 -0600
Subject: General: Web sign-up

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Dear Listers,

I remember seeing posts over the years regarding web-based equipment
sign-up systems, but didn't save any of them. Can anyone direct me to
an archived discussion thread, or have current suggestions on a very
simple and easy way to set this up? I want to put this under a dept.
web page on our intranet only, so security is not a major issue. All I
want is a simple calendar that users can view and sign up for time
slots. No retribution if time not used, but must require user's name so
that they can be tracked down for questioning if needed. I have no
experience in HTML programming but am not afraid to learn.

Thanks as always,
Karen

--
Karen S. Zaruba kszaruba-at-mmm.com
3M Company, St. Paul, MN






From: Karen Zaruba :      kszaruba-at-MMM.COM
Date: Tue, 29 Feb 2000 16:45:18 -0600
Subject: General: Video Printers

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Any suggestions on a really good video printer to include with an SEM
purchase? We've had trouble over the years with the standard roll
printers getting paper jams, which required sending out for servicing
(i.e. lots of down time). We're willing to sacrifice some speed for
something reliable. Are there "fast" ( { 1 min. per print) video
printers that use the cut sheet paper?

Thanks,
Karen

P.S. vendors please respond by E-mail (not phone) or dare to face my
wrath.
--
Karen S. Zaruba kszaruba-at-mmm.com
3M Company, St. Paul, MN






From: Barbara Plowman :      Bplowman-at-sfmail.dental.uop.edu
Date: Tue, 29 Feb 2000 15:09:31 -0800
Subject: subcribe

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From: Ekstrom, Harry :      harry.ekstrom-at-honeywell.com
Date: Tue, 29 Feb 2000 17:11:10 -0700
Subject: Tracor Northern 5500 EDX Units

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We have 2 Tracor Northern/Noran TN 5500 EDX systems that need a new home. We
are looking for a possible trade for comparable priced equipment/software.
If anyone is interested, please contact me directly.

Harry Ekstrom
Honeywell Materials Laboratory
Phoenix, AZ
602-231-2744





From: David Henriks :      Henriks-at-CompuServe.COM
Date: Tue, 29 Feb 2000 19:59:14 -0500
Subject: TEM: Tripod Polisher Workshop - Date

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Sorry for the inconvenience, but we mistakenly scheduled our Tripod
Polisher Workshop over Memorial Day Weekend. While I'm sure that you
would all love to spend your holiday weekend here with us, we felt it
necessary to move the workshop to the following week. The workshop is now
scheduled for June 2- 3, 2000 in San Clemente, CA.

For those of you who have already registered, you should have already been
contacted and given the option of re-booking for the new date or canceling
your reservation. Again, I apologize for the error. Certainly, if anyone
wishes to come and visit us over the Memorial Day Weekend, we will welcome
you with open arms - but you'll have to wait until the following weekend
for the workshop!

Best regards-

David
Writing at 5:12:19 PM on 02/29/2000

***************************************************************************
************************

David Henriks
Vice President TEL: 800-728-2233
(toll free in the USA)
South Bay Technology, Inc. +1-949-492-2600
1120 Via Callejon FAX: +1-949-492-1499
San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com

***************************************************************************
************************

} } } } } Please visit us at http://www.southbaytech.com { { { { {

Manufacturers of precision sample preparation equipment and supplies for
metallography, crystallography and electron microscopy.




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