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From: Richard Beanland +44 1327 356363 :      richard.beanland-at-gecm.com
Date: Tue, 01 Jul 1997 14:00:21 +0000 (GMT)
Subject: Re: Colour changes in Si

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On a related subject, has anyone else noticed that Si samples get darker when
you start to ion mill them? I usually get TEM cross-sections down to ~8-10
um, and when the sample goes in the mill it's an amber/red colour (from the
rather poor light shining through it). Ten minutes later it's almost opaque.
Maybe it's just the surface roughness increasing; or perhaps the effect of an
amorphous layer?
And while I'm on the subject of looking through samples in the ion mill, has
anyone come up with a way of seeing how thick GaAs or InP are while milling?
GaAs seems to become transparent at about 1 um (ish), and InP always looks
dark no matter what. A real pain when setting the timer!

Cheers,

Richard Beanland
GMMT Ltd.,
Caswell,
Towcester,
Northants NN12 8EQ
UK





From: Larry Stoter :      LPS-at-teknesis.demon.co.uk
Date: Tue, 1 Jul 1997 07:42:22 +0100
Subject: Re: Hydrogels-sample preparation for SEM, AFM

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} Recently we were asked to analyze the uniformity of hydrogel coating on the
} surface of the 100 micron particles. The hydrogel consists of 10% bifunctional
} polyacrylate and some additives. The rest, of course, is water. We froze the
} samples in liquid N2, and sublimed frozen water using freeze dryer. After SEM
} observation, teh coatings looked collapsed anyway. This is not what our
} customer wants. AFM analysis of the obtained samples yields similar
} results. In
} addition, it is compounded by the fact that the particles are spherical
} and the
} curvature really throws off the small magnificAtion images. We recommended the
} customer a liquid Tapping Mode AFM (we are not eqiupped with it). Is there any
} other way we can prepare and analyze the surface morphology of these samples?
} This is my first time addressing the ListServer, but there is always time for
} first.
} Thanks for possible help,
} Alex Mejiritski
} Ph. D. Student
} Center for Photochemical Sciences
} Bowling Green State University
} Bowling Green , Ohio 43402
} (419) 372-7830
}

How about an Environmental SEM? You could look at the sample in a fully
hydrated state, then slowly dehydrate, to reveal sub-surface structure.

Regards,
Larry Stoter






From: Jim Darley :      jim-at-proscitech.com.au
Date: Tue, 1 Jul 1997 14:20:36 +1000
Subject: Re: LM: length standard reply

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Ed is quite right, one space plus one line. If possible, the accuracy is
improved if many spaces/lines and are included. A stage micrometer could be
used to establish the magnification and for a given lens combination that
figure would be "permanent". For some uses it could be most useful to then
establish the length of the negative in micrometers and use that as a
standard, but some people would prefer large latex spheres incorporated
with the specimens. For low power SEM and reflected LM there is a 0.01mm
graduated scale available for calibration of those instruments (See Pelco
or ProSciTech).
Jim Darley

ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313
Great microscopy catalogue, 350+ Links, MSDS
************************ http://www.proscitech.com.au
} } } Can anyone suggest what to use as a length standard with a 100x
} } } objective? I intend to print out an image of this standard when I
print
} } } images of samples, in order to determine final magnification. I am a
little
} } } concerned with the use of my micrometer slide (10 microns between
} } } graduations) because the width of the individual lines is significant
} } } compared to the distance being measured between lines. Also there is
} } } no tolerance stated for the separation between the lines. I can think
of
} } } using calibrated latex beads suspended in water, but the refractive
index
} } } difference between latex and the aqueous medium (or air) causes a lot
of
} } } problems. Are there other standards I can use?
} } }
} } } Thanks
} } } Richard
} } } Richard_Thrift-at-Depotech.com
} }
} }
} } Richard,
} }
} } I was under the impression that micrometer slides were meant to measure
} } from one side of a line to the corresponding side on the adjacent line,
} } not between the lines. Have I been mis-led?
} } I thought the line thickness is accounted for in this manner.
} }
} } } | } | not | { } |
} }
} } Cheers
} } ed
} }
} } Edward J. Basgall, PhD
} } The Pennsylvania State University
} } Surface Chemistry Group ejb11-at-psu.edu
} } Materials Research Institute Building Ph:
814-865-0493
} } University Park, PA 16802-7003 FAX: 814-863-0618
} }
} }
}
}




From: gradice-at-richmond.edu (Gary Radice)
Date: Tue, 1 Jul 1997 10:21:42 -0400
Subject: test specimen for toy microscopes

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God help me, I had another idea....

What about a piece of black and and white photographic negative film for a
cheap test specimen? On a good microscope, the silver grains appear with
sharp edges. I don't have a toy microscope around to compare. But film is
widely available and cheap, dry, and you don't even need a microscope slide
or coverslip. No regular pattern to detect barrel or pincushion distortion,
but these probably wouldn't matter to most users anyway.

BTW, someone around here made up a bunch of cheap stage micrometers by
photographing a metric ruler from a distance with a 35 mm camera. With the
reduction, the one mm divisions on the scale are 100 micrometers apart on
the negative. These were then just cut up and glued to microscope slides.
They are a little fuzzy but if you measure center to center of each line it
is close enough. Perhaps one could photograph a grid to make a test
specimen for distortion.

Gary Radice, Associate Professor gradice-at-richmond.edu
Department of Biology 804-289-8107 (voice)
University of Richmond VA 23173 804-289-8233 (FAX)






From: Jim Darley :      jim-at-proscitech.com.au
Date: Tue, 1 Jul 1997 14:49:15 +1000
Subject: Re: Images of Algae and Sea Weeds

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David - use the find function on your browser and search for "Protist" on
our link page. That link takes you a sites at the Uni of Montreol which has
the very things you are looking for.
Cheers
Jim Darley

ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313
Great microscopy catalogue, 350+ Links, MSDS
************************ http://www.proscitech.com.au

----------
} From: David Webb {davehawaiiedu-at-msn.com}
} To: Microscopy-at-sparc5.microscopy.com
} Subject: Images of Algae and Sea Weeds
} Date: Monday, 30 June 1997 5:12
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} I am looking for all types of images (light & EM) which deal with algae,
} especially sea weeds. I plan to teach a course in which I will compare
and
} contrast vegetative and reproductive adaptations of algae with land
plants.
} The emphasis will be at the light microscope level, but SEM and TEM
images
} which deal with major aspects of form and function would be welcome.




From: Jim Darley :      jim-at-proscitech.com.au
Date: Tue, 1 Jul 1997 14:43:52 +1000
Subject: Re: Cleaning EM grids

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Cleaning grids? Go your hardest if you have nothing better to do:
Collect grids in a 20 or 30ml glass vial. Soak them in chloroform, give one
change in chloroform, pour off solvent, replace with ethanol or acetone,
replace with water, add drops of detergent, ultra sonicate for a short
time. Rinse with water, etch for a moment in weak acid (10% acetic or 0.2N
HCl) rinse in distilled water. Pour water off. Use washbottle with water
(or for faster drying ethanol) to flush grids into a Petrie dish lined
with filterpaper; pipette off excess fluid. Sit dish in an incubator until
dry. Thousands of grids can be treated in one batch but the joy is in
sorting the grids and throwing the bad once out.
We sell standard copper grids at A$10/vial/100; which is a little over
US$7. In a labour-costly country, with the odd exception, cleaning grids
is a waste of time.
Cheers
Jim Darley

ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313
Great microscopy catalogue, 400+ Links, MSDS
************************ http://www.proscitech.com.au



} I don't know about the first, I've never had really good results from
} cleaning off coated grids, but as to the second; in the absence of a Glow
} Discharge apparatus you might get some result out of using a Zerostat
} Antistatic Pistol. It is not so reliable as Glow Discharge and takes some
} practice to obtain neutral charge grids.
}
} They are still available as far as I know. We bought one recently for
} zapping our Balance after frustrating sessions of attempting to weigh out
} powders which were flying all over the bench. It cured that.
}
} If you want to see if it works before buying, find yourself a Black Vinyl
} Record collector and borrow one. They often use them for discharging
static
} on Discs before playing. I used one myself before going over to CD's many
} years ago.
} Regards
} Stephen Griffiths
}
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}
} Stephen Griffiths e-mail:- s.griffiths-at-ucl.ac.uk
} Visual Science Department Phone:- 0171 608 6914
} Institute of Ophthalmology Fax:- 0171 608 6850
} Bath Street, London. EC1V 9EL
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}
}
} } Just wondering about a couple of things.
} } First the different ways there are to clean grids that have a
} } film (2% parlodion), and a carbon coat, but no sample.
} } Second the best way to reduce or eliminate static on coated grids,
} } without using a glow discharge apparatus.
} } Thanks,
} } Maya Moody
}




From: Jim Darley :      jim-at-proscitech.com.au
Date: Tue, 1 Jul 1997 14:04:29 +1000
Subject: Re: Cleaning grids (Zerostat)

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Dear Joyce:

We still have a "bunch" available. See our on-line catalogue.
Jim Darley

ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313
Great microscopy catalogue, 400+ Links, MSDS
************************ http://www.proscitech.com.au
----------
} From: joyce craig {bafpjec-at-csu.edu}
} To: Listserver {Microscopy-at-sparc5.microscopy.com}
} Subject: Cleaning grids (Zerostat)
} Date: Tuesday, 1 July 1997 15:18
}
} ------------------------------------------------------------------------
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} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Does anyone know where I can purchase a Zerostat antistatic pistol? My
} old source no longer carries them, and I find them really useful in dry
} sectioning in low humidity (not a problem in Chicago in the summer, but
} in the winter...) and in keeping grids from bouncing onto the lid of the
} Petrie dish.
} Cheers from
} Joyce Craig
} Chicago State University




From: oshel-at-ux1.cso.uiuc.edu (Philip Oshel)
Date: Tue, 1 Jul 1997 08:02:48 -0500
Subject: RE: Diamond objectives? Coffee break discussion

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From Mark Lund:
}
} ---
}
} Being the token optical engineer on this list makes me the token lens
} designer, too :) Although I have never designed an immersion type
} microscope objective the general trend is that as you go up in index
} of refraction the job gets easier. A high index "glass" that won't
} etch should make the lens design practical. Of course the high index
} of diamond should make it possible to get higher NA's

But diamond has a high dispersion relative to quartz (0.044 vs 0.013).
Wouldn't this make correcting for chromatic aberration difficult?

How about corundum? RI = 1.762 -1.77, dispersion = 0.018

On a separate note: as the "token optical engineer", have you been
following the thread about microscopes for kids and
lower-cost-but-still-quality ones for amateurs?

Phil


} Sic Hoc Legere Scis Nimium Eruditionis Habes {
Philip Oshel
Station A
PO Box 5037
Champaign, IL 61825-5037
(217) 355-1143
oshel-at-ux1.cso.uiuc.edu
*** looking for a job again ******************







From: DDKJoe-at-aol.com
Date: Tue, 1 Jul 1997 07:45:07 -0400 (EDT)
Subject: Re: Diamond objectives? Coffee break discussion

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The index of refraction of diamond is 2.4173. I don't know as much about the
theory of microscope optics as I should but I do know that this plays a role.

Joe Tabeling
Delaware Diamond Knives
3825 Lancaster Pike
Wilmington, DE 19805
302-999-7476




From: Nestor J. Zaluzec-Argonne Nat. Lab. :      ZALUZEC-at-aaem.amc.anl.gov
Date: Tue, 1 Jul 1997 11:19:40 -0500 (CDT)
Subject: Microscopy Back On-Line -- ???

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Colleagues;

The server was down for nearly a full day while I
replaced the main drive. Expect a hic-cup or two
while things settle in.

The subscribe/unsubscribe functions should (?) be
functioning tomorrow AM. For those of you that
have tried to unsubscribe you should be "off" the
list tomorrow.

I apologize for the inconvenience...

Nestor
Your Friendly and Blurry Eyed Neighborhood SysOp


End of returned message





From: Christoph Guenther :      guenther-at-141.42.1.11
Date: Tue, 01 Jul 1997 16:09:35 +0200
Subject: cryo-sectioning

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Hallo!


We want to do immunohistochemistry on human heart muscle tissue cryosections.
Which embedding medium should be used for cryosectioning on a Leica cm
3000 kryostat for light microscopy?
Are there simple recipies and procedures (cryoprotection...)?
Which freezing techniques should be used? We get the tissue in the
operating theatre which freezing technique can be used directly there? I
sthere a freezing technique for both, cryosectioning and RNA-preparation of
the same tissue-block.

Thank you all for your answers

Christoph Guenther
Klinikum der Charite der Humboldt Universitaet
Ziegelstrasse 5-7
10117 Berlin /Germany
Tel.: +49 30 2802 6327
Fax + 49 30 2802 6608




From: vhacopian-at-wellesley.edu (Vachik Hacopian)
Date: Tue, 01 Jul 1997 12:49:48 -0500
Subject: Re: Toy Microscopes, Resolution & Education

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Caroline Schooley wrote:

} There's a new CD-ROM (Windows & Mac) out
} from a reputable source (Center for Bioengineering, U. of Washington,
} Seattle) that may do some good (tho I haven't had a chance to review it yet
} for the Project MICRO bibliography). Microscopy-Tutor, Lippincott-Raven,
} ISBN 0-7817-1217-3, $195.00. http://www/lrpub.com, 800-777-2295.

The URL for Microscopy-Tutor is "http://www.lrpub.com/media/m1208.htm".

Here's an advertising blurb from the web site.

==============================================
Microscopy-Tutor, CD-ROM for PC and Macintosh, with Two-Color Insert, by
The Department of Laboratory Medicine and the Center for Bioengineering,
University of Washington, Seattle, WA.

This interactive computer program guides students of biology and the health
professions through the basic concepts of bright field light microscopy,
developing and refining the user's knowledge by providing a more active
role in learning. Simple, approachable, and largely qualitative,
Microscopy-Tutor uses three-dimensional animation to simulate a microscope
with an integrated illuminator and adjustable field diaphragm. The CD-ROM's
animated diagrams are more accurate than those found in most
university-level texts, but are also easy-to-understand because key
concepts and equations are presented visually rather than symbolically.
Since using Microscopy-Tutor feels more like operating a microscope than a
computer it's successful in presenting changes in dynamic processes that
occur during alignment and use of the microscope. Rather than explaining
what happens through the use of words and pictures, the user -- who learns
by doing -- can see what happens and better understand the implications. In
some sections two animated perspectives are synchronized, allowing the
viewer to change microscope settings and simultaneously see the resulting
image. Interactive quizzes test the user's knowledge and accent areas for
improvement.
==============================================

I have no experience with this software.

Vachik Hacopian






From: Dr. Mark W. Lund :      lundm-at-acousb.byu.edu
Date: Tue, 01 Jul 1997 10:44:27 MST/MDT
Subject: RE: Toy Microscopes, Resolutions and Aberrations

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The information below is slightly wrong. Not a big deal, but in
the interest of science I thought I would set things slightly
straighter :)

Azriel Gorski wrote tht McCrone and McCrone wrote:

} Lenses have several types of aberrations which will cause the loss of
} detail unless corrected for. Toy microscopes are not.
}
} Spherical aberration - "is especially apparent in lenses having sperical
} surfaces. Light paths near the center of the lens focus at different
} points compared to light paths near the periphery."


The name "spherical aberration" has historical roots in astronomy where
a spherical mirror has this aberration but a parabolic mirror does not.
In some instances a spherical mirror will have no spherical aberration
wheras a parabolic mirror will have maximum spherical aberration.
The aberration really has nothing to do with the spherical surfaces
of lenses. The confusion comes because making a surface "aspheric"
can cure spherical aberration in many instances.


} Field curvature - "is a natural result of using lenses with curved
} surfaces. The image plane produced by such lenses will be curved. This
} kind of image occurs in microscopy unless plano (flat-field) objectives
} are used."

Actually, field curvature is a natural result of the geometry of the
real world. Since an object at the edge of the field of view is
farther from the lens "center" it will tend to be focussed closer to the lens
than an object on axis. This naturally leads to field curvature unless
the designer makes the lens weaker for off axis points. It has nothing
to do with the lenses being curved. The confusion comes from the formula
for Petzval curvature, which has lens powers in it.


To the amateurs - welcome! We need you to keep us fresh and excited in
what we do.

Shalom from Utah

mark

/signature






From: Robert Underwood :      underwoo-at-u.washington.edu
Date: Tue, 1 Jul 1997 10:01:22 -0700 (PDT)
Subject: Re: Digoxigenin labeled in situ's

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Hello Tom,

The alk phos is more sensitive because it will continue to reduce
substrate long after peroxidase has quit.

Bob
Morphology Core


On Fri, 27 Jun 1997, Tom Phillips wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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} -----------------------------------------------------------------------.
}
} We are starting some in situ work with digoxigenin labeled probes. A lot
} of the kits and studies seem to use Alkaline Phosphatase labeled antibodies
} to detect the digoxigenin. I am curious why they seem to prefer Alk Phos
} over peroxidase coupled antibodies which are the more standard
} immunocytochemical choice. Anybody have any thoughts on this?
}
}
} Thomas E. Phillips, Ph.D.
} Associate Professor of Biological Sciences
} Director, Molecular Cytology Core Facility
} 3 Tucker Hall
} University of Missouri
} Columbia, MO 65211
} (573)-882-4712 (voice)
} (573)-882-0123 (fax)
}
}
}





From: Frank Karl :      fskarl-at-goodyear.com
Date: Tue, 01 Jul 1997 13:22:21 -0400
Subject: Diamond objectives

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Wow!

Talk about a step backwards... Historically, some objectives were made
with different gems in the 1800's (I believe, I don't have my history
references at work), but they suffered from several problems. First there
are problems with the optical clarity of the gem, color in the case of
rubys and garnets, polishing the surfaces to the optimum shape and of course,
cost. In the case of polarized light, the double refractive index stones
were disastrous. This development did not lasted very long as it was too
impractical. Also remember, an immersion system requires more than oiling
the front lens of the objective. The condenser should be oiled to the
slide as well as the objective. In these systems the resolution is limited
by the lowest refractive index, which for a diamond condenser and objective
could be the immersion oil.

Good luck and have fun!


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

These opinions are mine alone and have no relationship to my employer.
Thank you.

Frank Karl fskarl-at-goodyear.com
Goodyear Tire & Rubber Co. Voice: 330-796-7818
Analytical Services Dept. 415B Fax: 330-796-3304
142 Goodyear Blvd.
Akron, OH 44305
USA

They that give up essential liberty to obtain a little temporary safety
deserve neither liberty or safety. Benjamin Franklin




From: Ray Hicks :      rh208-at-cus.cam.ac.uk
Date: Tue, 1 Jul 1997 10:53:00 +0100
Subject: Re: Cleaning grids (Zerostat)

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I think Aldrich still sell them, catalogue number: Z10,881-2, I've only got
an old catalogue, and Aldrich might be your old source, so this might not
be too useful.

Ray

At 10:18 pm -0700 30/6/97, joyce craig wrote:
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Ray Hicks
________________________________________________________________________
|University of Cambridge |Tel 01223 330149 |
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|CB2 |ftp server ftp://131.111.80.78 |
|UK | |
|_________________________________|_____________________________________|






From: Keith Ryan :      KPR-at-wpo.nerc.ac.uk
Date: Tue, 01 Jul 1997 08:39:44 +0000
Subject: Re: Recommendations on freeze dryers -Reply

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I have looked at the discussion on recrystallization with interest. As I am an agreeable fellow, I would say that you are both correct!

It is true that amorphously vitrified pure water will devtrify at about -130 C or lower, depending partly on the rates of cooling and rewarming. The
other extreme was described by MacKemzie AP (1981) Modellling the ultra-rapid freezing of cells and tissues. In: Microprobe analysis of Biological
Specimens (eds) Hutchinson TE & Somlyo AP. Academic Press, NewYork, London, 397-421. In this work, which used starches and gels, some specimens could
be rewarmed to about -6 C before crystal growth took place (i.e. in the higher molecular weight substances).

I published one micrograph in Scanning Microsc. 6, 715-743 (KP Ryan, Cryofixation of tissues for electron microscopy: a review....) in 1992 which
showed some fish spleen tissue elements stored at -60 for 48 hopurs which showed no observable signs of crystal growth. Other results (some published
only in my thesis) showed that the samwe tissue could be stored at -40 for some days but after 8 days there were signs of crystal growth in the
extracellular fluid (more 'watery'?).

This is an interesting aspect of 'cryo' and deserves more attention by someone. I only did a piece of this to validate the 48 h at -80 C freeze
substitution that I used. It should not induce crystal growth. The same f/s has been done at -50 C without apparent crystal development (but not by
myself). In tthat case I presume the substitution got to the frozebnwater before it had time to show growth in the crystals

Keith Ryan
Plymouth Marine Lab., UK




From: yuhui xu :      Yuhui=Xu%RES%DFCI-at-EYE.DFCI.HARVARD.EDU
Date: Tue, 1 Jul 97 13:51:05 EDT
Subject: Where to Buy Filaments

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Dear colleagues:
All of our scopes are JEOL made and we have been using filaments made by
JEOL. The engineers from JEOL also recommend us to use their parts. As far as
I know, however, there are companies which also sell filaments. Could anyone
of you give me some information as to which source is better than the others?
Thank you very much.
Regards,
Yuhui




From: Dr. Mark W. Lund :      lundm-at-acousb.byu.edu
Date: Tue, 01 Jul 1997 12:04:44 MST/MDT
Subject: RE: Diamond objectives? Coffee break discussion

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Philip Oshel wrote:

} } From Mark Lund:
} }
} } ---
} }
} } Being the token optical engineer on this list makes me the token lens
} } designer, too :) Although I have never designed an immersion type
} } microscope objective the general trend is that as you go up in index
} } of refraction the job gets easier. A high index "glass" that won't
} } etch should make the lens design practical. Of course the high index
} } of diamond should make it possible to get higher NA's
}
.But diamond has a high dispersion relative to quartz (0.044 vs 0.013).
} Wouldn't this make correcting for chromatic aberration difficult?
}
} How about corundum? RI = 1.762 -1.77, dispersion = 0.018
}
} On a separate note: as the "token optical engineer", have you been
} following the thread about microscopes for kids and
} lower-cost-but-still-quality ones for amateurs?

First: sorry about my first post. My computer decided that it had
more important things to think about and while I was trying to
get its attention the message posted before I could finish it.

The important thing is not the dispersion--many glasses have higher
dispersion than diamond, but that the dispersion is much higher
than you would expect for a refractive index of 2.4. Not only
is the dispersion anomalous, but the partial dispersions are all
well off the normal "glass line." This means that making a
lens where one of the elements is diamond would be much easier
than otherwise. Not only would correcting the color be easier
but correcting residual color (i.e. making the lens apochromatic)
would be easier. Also, for an immersion lens (with the right
index matching oil) you could probably get an NA greater than
2.0, maybe even as high as 2.3. Of course the condensor and
slide would also need to be high index for this to be useful :(

It looks like a fun idea, let me know if you need someone
to design one!

Sapphire would be the perfect first element except for its
natural birefringence--it would come out astigmatic on axis.


best regards
mark

Mark W. Lund, PhD
Director } } Soft X-ray Web page http://www.moxtek.com { {
MOXTEK, Inc.
452 West 1260 North
Orem UT 84057 801-225-0930 FAX 801-221-1121
lundm-at-xray.byu.edu

"The state is good at simple tasks, like killing people and seizing their
wealth. It has far more trouble reaching inside individuals and making them
good." Doug Bandow








From: Ultratecex-at-aol.com
Date: Tue, 1 Jul 1997 14:30:31 -0400 (EDT)
Subject: Annular Cutting Machine -- Re:Cutting Titanium Implants

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

Marcello,

With regard to cutting undecalcified bone samples (with and without
implants), a lot of people have found the best method of achieving high
straightness, low damage, thin cuts is with an annular diamond blade.

This blade involved in this process is held in a precision chuck and mounted
at high tension, to give very little lateral movement, thus producing the
high quality of cut. There are many exponents of the method in the hard
tissue & dental research fields.

I'd be pleased to send you a relevant technical article on the use of this
system for this application and also further information on the commercial
version of the annular cutting machine, the 'Microslice 2'. Please send me
your address details so I may get the information through to you.

(Note to Group: This product is sold by us -- Please accept this as a
notification of our commercial interest in this product)

I look forward to hearing from you.

Best Regards

Tim Hazeldine
Product Manager, ULTRA TEC Mfg., Inc.
...............................................................the cutting
and polishing specialists
1025 E. Chestnut Avenue
Santa Ana
CA 92701 - 6425
USA
Tel. +1 714 542 0608 Fax. +1 714 542 0627
e-mail. info-at-ultratecusa.com or ultratecex-at-aol.com
..............................................................................
.....................................




From: William Tivol :      tivol-at-wadsworth.org
Date: Tue, 1 Jul 1997 14:36:47 -0400 (EDT)
Subject: Re: Diamond objectives? Coffee break discussion

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

Dear Joe,

} The dispersion is 0.044.
}
In units of per nanometer? In any case, I still don't know what
effect this might have on the practicality of using diamond lenses. If
the illumination were single-wavelength, there would be no problem.

} Fabrication problems?
}
Yes, but a colleague suggests that one can use diamond powder as
a grinding agent. You at DDK would know a lot more than I. On the plus
side, diamond lenses would be very hard to scratch.
Yours,
Bill Tivol





From: DANIEL SCHWAB :      DSCHWAB-at-OPIE.BGSU.EDU
Date: Tue, 01 Jul 1997 14:58:43 -0500 (EST)
Subject: EDX Interpretation

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

Hello everybody,

Recently we did an SEM/EDX analysis that gave rather baffling results.
After some thought we came up with what seemed to be a plausible
explanation, however, the people for whom the analysis was done remained
skeptical. Therefore I am writing the Listserver for a second opinion.
The sample consisted of irregular shaped alumino-silicate particles roughly
2-5 microns is size. The specimen was uncoated resulting in some charging.
Analysis (windowless) at 1000x mag. resulted in a carbon peak much larger
than the Al or Si peaks. At 5000x the carbon peak was smaller; i.e.,
comparable to the Al and Si. Using a spot mode on individual particles, no
carbon was detected. However, spot mode analysis after lowering the
accelerating voltage from 20 to 10 kv, showed a small carbon peak.
The only explanation we could envision to fit these results was a thin
carbonaceous coating on the particle surface. Is this a plausible
explanation or we missing something? Does anyone have an alternative
suggestion? Any help would be appreciated.

Dan Schwab
Center for Microscopy and Microanalysis
Bowling Green State University
Bowling Green, OH





From: corwinl-at-pt.cyanamid.com
Date: Tue, 01 Jul 1997 15:29 -0500 (EST)
Subject: Re: LM: length standard

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

Leica (and probably other manufacturers) sells NIST (US) or NPL (UK)
traceable stage micrometers, certified to tenths of a micron.
Measurements are from centers of the lines. They're not cheap.

If you want to do your own certification, start with some regular fine
grid, perhaps a dime store holograph, measure it against some standard
you trust, and figure out the spacing.




From: Buddy Steffens :      steffens-at-calc.vet.uga.edu
Date: Tue, 1 Jul 1997 16:33:51 EST
Subject: Re: Cleaning grids (Zerostat)

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

To Joyce Craig:

I have just today found a company that supplies the Zerostat
antistatic pistol in the U.S.

Bellex International
Voice: (302) 761-9885
FAX: (302) 761-9896

The company is located in Wilmington, DE. Ask for Deborah Hunt.


-=W.L. Steffens=-
Department of Veterinary Pathology
College of Veterinary Medicine
University of Georgia
Athens, GA 30602 USA
http://www.vet.uga.edu/vpp/wls/steffens.html




From: Wil Bigelow :      bigelow-at-engin.umich.edu
Date: Tue, 1 Jul 1997 12:09:20 -0400
Subject: Vac Dessicator storage

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

There are at least two problems involved in storing objects (specimens,
pole pieces, specimen holders, and other things commonly stored in
dessicators) in a plastic vacuum dessicator under vacuum: (1) the
dessicator may have a small leak, and over time come back up to atmospheric
pressure, thus exposing the objects to ordinary air, (2) under the
influence of the vacuum plasticizers may bleed out of the plastic from
which the desiccator is made and find their way onto the stored objects, as
noted by others.

Both of these problems can be minimized quite simply, however, by filling
the dessicator to atmospheric pressure with a dry, inert gas, rather than
leaving it evacuated. For most purposes dry nitrogen would be a
satisfactory gas to use here, although helium or argon might be preferred
for storing some reactive materials. If the gas is purchased in a high
pressure tank it is necessary to be sure that it is oil-free, otherwise the
fill-gas may carry oil vapors onto the stuff you are storing inside the
dessicator, thereby defeating the purpose of the whole operation, a matter
discussed on p. 64 of my book 'Vacuum Methods in Electron Microscopy'.

The vapor pressure of water at the temperature of liquid nitrogen is in the
realm of 10-20 Torr, and the vapor pressure of most oils is even lower, and
so the gas that is constantly boiling off each container of liquid nitrogen
is about as clean and dry as any you can get. As described on p. 65 of Vac
Meth in EM, this dry nitrogen can be collected and used to fill vacuum
apparatus rather simply. Fit a one-hole stopper into the LN2 storage flask
and connect it to the inlet valve of the vacuum container with
non-collapsable, flexible tubing (ordinary polyethylene tubing works well).
Attach a large, collapsable plastic beach ball to a Tee joint in this
tubing with a length of soft, surgical-rubber tubing, and make a clean slit
in this surgical tubing about 100 mm long with a sharp razor blade or
scalpel. Ordinarily this slit will close thghtly enough so that the
nitrogen gas evolved from the storage container will be directed into the
beach ball; however, if the ball becomes full the slit will serve as a
primative pressure-release valve by opening slightly and allowing the gas
to escape so that there is no danger of over-pressurization. When the
inlet valve to the evacuated chamber is opened, the nitrogen will flow into
the chamber only under the influence of atmospheric pressure acting on the
collapsable beach ball, and so there is no danger of exceeding atmospheric
pressure in the chamber. An ordinary beach ball will hold enough gas to
fill most dessicators several times.

If an oil-sealed rotary vane pump is used to evacuate the storage chamber
then some care must be exercised to avoid having oil vapours backstream
from the pump into the chamber. To avoid this, it is important not to pump
the chamber down below the range of viscous flow (i.e. below about 0.1 Torr
- see p.29 of Vac Meth in EM). If this is not considered to be a
sufficient vacuum to remove as much atmospheric gas as desired, then the
container can be filled with the dry gas, pumped out and filled again a
couple of times. Each time the container is pumped out some 90% of the
existing gas molecules are removed, and so two or three flushings should
leave only an insignificant trace of the original atmospheric gases.

Wilbur C. Bigelow, Prof. Emeritus
Materials Sci. & Engr., University of Michigan
Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu;
Fx:313-763-4788; Ph:313-764-3321






From: Dr. Gordon Warren :      root-at-rangers.tamu.edu
Date: Tue, 1 Jul 1997 16:32:14 -0500
Subject: Re: cryo-sectioning

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

On Jul 1, 4:09pm, Christoph Guenther wrote:
} We want to do immunohistochemistry on human heart muscle tissue cryosections.
} Which embedding medium should be used for cryosectioning on a Leica cm
} 3000 kryostat for light microscopy?
} Are there simple recipies and procedures (cryoprotection...)?
} Which freezing techniques should be used? We get the tissue in the
} operating theatre which freezing technique can be used directly there? I
} sthere a freezing technique for both, cryosectioning and RNA-preparation of
} the same tissue-block.

In skeletal muscle, we embed in OCT compound (Miles, Inc.) and plunge freeze in
melting isopentane. To avoid freeze artifact, go with smaller pieces of tissue
( { 20-30 mg). Also, keep the specimen in the melting isopentane only for 8-12
sec otherwise sample cracking occurs. For more detail, see the "Color Atlas of
Muscle Histochemistry" by R.A. Brumback and R.W. Leech (PSG Publishing Co.,
1984).


--
Gordon L. Warren, Ph.D.
Research Scientist
Muscle Biology Laboratory
158 Read Building
Texas A&M University
College Station, TX 77843-4243
Fax (409) 862-4808
Phone (409) 862-4809 office
e-mail root-at-rangers.tamu.edu




From: Don Chernoff at ASM :      asm-at-indy.net
Date: Tue, 01 Jul 97 10:32:04 -0500
Subject: Re: Hydrogels-imaging.

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

Alex Mejiritski asked about imaging hydrogels.

He remarked that freeze-drying (for SEM) produced collapsed coatings and
that AFM (imaging in air) produced the same result.

} We recommended the
} customer a liquid Tapping Mode AFM (we are not eqiupped with it).
-- I am happy to say that ASM is equipped to do exactly this. We have
had good success imaging wet samples.

Don Chernoff
Analytical Services Division



Advanced Surface Microscopy, Inc. E-Mail: asm-at-indy.net
6009 KNYGHTON RD. Voice: 317-251-1364
INDIANAPOLIS IN 46220 Toll free: 800-374-8557 (in USA)
web: http://www.a1.com/asm Fax: 317-254-8690
(If you experience difficulty in accessing our website,
note that the web address uses numeral "1" in "a1")







From: Linda Barthel :      barthel-at-umich.edu
Date: Tue, 1 Jul 1997 18:59:51 -0400 (EDT)
Subject: Re: cryo-sectioning

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

We use a 1:2 mixture of OCT with our cryoprotectant buffer (PO4 + 20%
sucrose) freezing in isopentane cooled in liquid nitrogen. This gives us
a block consistency that allows for 3 um cryosections at -20 C, see
Barthel & Raymond, J. Histochem Cytochem, 1990, 38:1383-1388 or our web
site for a detailed protocol, http://www.personal.umich.edu/~praymond/
Others have used this combination with good success for both thin and
thick (+10 um) sections.

To avoid sample cracking we try to be sure that ratio of block size to
tissue size is large. It seems our tissue swells somewhat when it
freezes. If the block is not large enough then it cracks. Sometimes we
avoid the cracking by "quick" dipping the sample in the cooled isopentane
until it is completely frozen.

If anyone has other suggestions to avoid cracking that would be great. It
is a continuous battle with our samples. Seems some days are worse than
others.

For using the samples for RNA (I presume you mean in situs?) we treat the
samples no differently except that our reagents and labware (including
poly-L-lysine slides) are RNase free.

When collecting your sample from the OR is it necessary to directly freeze
there? Your best preservation would be if you placed your
sample in a bottle of fix and returned to the lab to continue the
processing.
Linda Barthel
Research Associate II
Department of Anatomy and Cell Biology
University of Michigan
lab (313) 764-7476
fax (313) 763-1166
barthel-at-umich.edu






From: Linda Barthel :      barthel-at-umich.edu
Date: Tue, 1 Jul 1997 18:59:51 -0400 (EDT)
Subject: Re: cryo-sectioning

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

We use a 1:2 mixture of OCT with our cryoprotectant buffer (PO4 + 20%
sucrose) freezing in isopentane cooled in liquid nitrogen. This gives us
a block consistency that allows for 3 um cryosections at -20 C, see
Barthel & Raymond, J. Histochem Cytochem, 1990, 38:1383-1388 or our web
site for a detailed protocol, http://www.personal.umich.edu/~praymond/
Others have used this combination with good success for both thin and
thick (+10 um) sections.

To avoid sample cracking we try to be sure that ratio of block size to
tissue size is large. It seems our tissue swells somewhat when it
freezes. If the block is not large enough then it cracks. Sometimes we
avoid the cracking by "quick" dipping the sample in the cooled isopentane
until it is completely frozen.

If anyone has other suggestions to avoid cracking that would be great. It
is a continuous battle with our samples. Seems some days are worse than
others.

For using the samples for RNA (I presume you mean in situs?) we treat the
samples no differently except that our reagents and labware (including
poly-L-lysine slides) are RNase free.

When collecting your sample from the OR is it necessary to directly freeze
there? Your best preservation would be if you placed your
sample in a bottle of fix and returned to the lab to continue the
processing.
Linda Barthel
Research Associate II
Department of Anatomy and Cell Biology
University of Michigan
lab (313) 764-7476
fax (313) 763-1166
barthel-at-umich.edu






From: Caroline Schooley :      schooley-at-mcn.org
Date: Tue, 1 Jul 1997 16:11:22 -0700
Subject: Re: test specimen for toy microscopes

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

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

Not just an idea, but two GREAT ideas! I knew that the free spirits on the
listserver would think of good stuff. Thanks. CS
}
} What about a piece of black and and white photographic negative film for a
} cheap test specimen? On a good microscope, the silver grains appear with
} sharp edges. I don't have a toy microscope around to compare. But film is
} widely available and cheap, dry, and you don't even need a microscope slide
} or coverslip. No regular pattern to detect barrel or pincushion distortion,
} but these probably wouldn't matter to most users anyway.
}
} BTW, someone around here made up a bunch of cheap stage micrometers by
} photographing a metric ruler from a distance with a 35 mm camera. With the
} reduction, the one mm divisions on the scale are 100 micrometers apart on
} the negative. These were then just cut up and glued to microscope slides.
} They are a little fuzzy but if you measure center to center of each line it
} is close enough. Perhaps one could photograph a grid to make a test
} specimen for distortion.
}
} Gary Radice, Associate Professor gradice-at-richmond.edu
} Department of Biology 804-289-8107 (voice)
} University of Richmond VA 23173 804-289-8233 (FAX)


Caroline Schooley
Educational Outreach Coordinator
Microscopy Society of America
Box 117, 45301 Caspar Point Road
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html
Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/






From: Garber, Charles A. :      cgarber-at-2spi.com
Date: Tue, 01 Jul 97 21:53:51 -0500
Subject: Where to Buy Filaments

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

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

Yuhui wrote:
==================================================
All of our scopes are JEOL made and we have been using filaments made by
JEOL. The engineers from JEOL also recommend us to use their parts. As far
as I know, however, there are companies which also sell filaments. Could
anyone of you give me some information as to which source is better than
the others?
===================================================
While new filaments are always an option, have you considered retipping your
left over filament bases? While not everyone would agree, there is a good
number of persons who would claim that a good retipped filament is
indistinguishable in its performance from a brand new one. Of course, not
all retipped filaments from different sources are created equal. But if
they work for you, a considerable amount of money can be saved.

Disclaimer: SPI Supplies offers a service to retip filaments as do others
such as some of our competitors such as Pella, EBS, PLANO, etc. so we would
all have a vested interest in seeing more people using retipped filaments
instead of new ones. You can find out more information about the retipping
of filaments from our website given below.

Chuck

===================================================
Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400
President 1-(800)-2424-SPI
SPI SUPPLIES FAX: 1-(610)-436-5755
PO BOX 656 e-mail: cgarber-at-2spi.com
West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com


Look for us!
############################
WWW: http://www.2spi.com
############################
=================================================




From: Beth Bray :      bbray-at-netside.com
Date: Tue, 1 Jul 1997 23:56:47 -0400
Subject: Sputter coating with aluminum

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

Hello to all,

I have a sample that I have labeled with gold tagged antibody and I want to
sputter coat it with aluminum for viewing in the SEM. We have a sputter
coater with a turbomolecular pump and an aluminum target. We tired to
sputter coat the sample using essentially the same "settings" used for
chromium, but could not see that any coating had occurred. Does anyone
have any ideas? Thanks in advance for any help any of you can give this
humble student.

VTY,
Beth Bray
bbray-at-netside.com




From: Jim Darley :      jim-at-proscitech.com.au
Date: Wed, 2 Jul 1997 14:47:11 +1000
Subject: Re: EDX Interpretation

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

Dan - Never trust the low energy peaks from a specimen that is charging.
Jim Darley

ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313
Great microscopy catalogue, 400+ Links, MSDS
************************ http://www.proscitech.com.au

} Recently we did an SEM/EDX analysis that gave rather baffling results.
} After some thought we came up with what seemed to be a plausible
} explanation, however, the people for whom the analysis was done remained
} skeptical. Therefore I am writing the Listserver for a second opinion.
} The sample consisted of irregular shaped alumino-silicate particles
roughly
} 2-5 microns is size. The specimen was uncoated resulting in some
charging.
} Analysis (windowless) at 1000x mag. resulted in a carbon peak much larger
} than the Al or Si peaks. At 5000x the carbon peak was smaller; i.e.,
} comparable to the Al and Si. Using a spot mode on individual particles,
no
} carbon was detected. However, spot mode analysis after lowering the
} accelerating voltage from 20 to 10 kv, showed a small carbon peak.
} The only explanation we could envision to fit these results was a thin
} carbonaceous coating on the particle surface. Is this a plausible
} explanation or we missing something? Does anyone have an alternative
} suggestion? Any help would be appreciated.
}
} Dan Schwab
} Center for Microscopy and Microanalysis
} Bowling Green State University
} Bowling Green, OH
}




From: chris gilpin :      cgilpin-at-fs1.sem.man.ac.uk
Date: Wed, 2 Jul 1997 09:24:35 BST
Subject: Re: Hydrogels-sample preparation for SEM, AFM

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



} How about an Environmental SEM? You could look at the sample in a fully
} hydrated state, then slowly dehydrate, to reveal sub-surface structure.
}
} Regards,
} Larry Stoter

We have successfully images hydrogel microspheres in the wet state
using ESEM.
See
Applications of the environmental scanning electron microscope to the
analysis of pharmaceutical formulations.
D'Emanuele A, Gilpin C Scanning 1996 Oct 18:7 522-7

Chris


Chris Gilpin
Biological Sciences Electron Microscope Unit
G452 Stopford Building
Oxford Road
Manchester
M13 9PT
phone +44 161 275 5170
fax +44 161 275 5171




From: Keith Ryan :      KPR-at-wpo.nerc.ac.uk
Date: Wed, 02 Jul 1997 10:24:14 +0000
Subject: Sputter coating with aluminum -Reply

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

If all else fails, can you put in a coating unit with a tungsten filament and evaporate aluminium wire/foil, from four sides in rotation?

Even better - 8 sides! (we have a very old unit that does this job but it has a specimen rotation arrangement).

Keith Ryan
Plymouth Marine Lab. UK




From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Wed, 2 Jul 1997 10:34:19 -0000
Subject: keeping a protozoa still

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

Is there a method of restricting a protozoa such as paramecium from
moving about? The protozoa solution I have is great for studying them,
but not if I want to look at the internal structure. Also what stain is
a good stain to bring out the details in paramecium such as the nucleus?

Finally how do you pronounce 'paramecium'?!!!

Conrad :)

------------------------------------------------------------------------
-----------
"Any sufficiently advanced technology is indistinguishable from magic"
----------------------------------------------------------- Arthur C
Clarke ----





From: EUGENE GORDON :      MEDJET-at-worldnet.att.net
Date: Tue, 1 Jul 1997 09:25:01 -0400
Subject: TEM preparation of human cornea

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

I am working with the human cornea and wish to find the optimal buffers for
glutaraldehyde and osmium tetroxide dilutions in the preparation of
specimens for TEM. Any suggestions or references would be greatly
appreciated.





From: Robert H. Olley :      R.H.Olley-at-reading.ac.uk
Date: Wed, 2 Jul 1997 11:38:55 +0100 (BST)
Subject: Re: Cleaning EM grids: KerBoom!?

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


} Cleaning grids? Go your hardest if you have nothing better to do:
} Collect grids in a 20 or 30ml glass vial. Soak them in chloroform, give one
} change in chloroform, pour off solvent, replace with ethanol or acetone,

Ethanol is better, if you store acetone and chloroform together in a waste
bottle there is a risk, as those two will react explosively under some
cirecumstances.

Also, after a final wash in aqueous media, if they are such as to remove
the natural oxide coating, the copper underneath can oxidize and go all
'orrible.

} In a labour-costly country, with the odd exception, cleaning grids
} is a waste of time.

I don't know if this applies to the USA, but in the UK we find that
non-standard washing up is too difficult to be left to untrained staff!

+------------------------------------------------------------------------+
| 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: ebs-at-ebsciences.com
Date: Wed, 02 Jul 1997 07:17:52 EST
Subject: Where to Buy Filaments

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

Dear fellow microscopists,

Yuhui Xu asked:
} All of our scopes are JEOL made and we have been using filaments made by
} JEOL. The engineers from JEOL also recommend us to use their parts. As far as
} I know, however, there are companies which also sell filaments. Could anyone
} of you give me some information as to which source is better than the others?

We (Energy Beam Sciences) have been manufacturing filaments for electron
microscopes for more than 25 years. I know that some third party filaments
are equal in quality to those purchased from the original equipment
manufacturer, and some are not. In shopping for filaments, it is important
to ask whether the filaments are vacuum-annealed (to minimize drift when
heated) and precentered *after* annealing.

It is also important to know that, although the original equipment
manufacturers all offer only one style of filament loop on their filament
bases, usually a "V" hairpin filament (eg Philips) or a "curved" loop (eg
JEOL), other filament loops can be mounted on the same bases to maximize
performance in one way or another. For example, a pointed filament can
dramatically increase brightness, while another loop configuration, such as
our trademark "AR" loop, will provide longer life.

Anyone interested in the details of these special loop configurations can
contact me directly back-channel.

Disclaimer: Energy Beam Sciences manufactures a wide range of tungsten
filaments for electron microscopes.

Best regards,
Steven E. Slap, Vice-President
********************************
Energy Beam Sciences, Inc.
Adding Brilliance To Your Vision
ebs-at-ebsciences.com
http://www.ebsciences.com/
********************************





From: Woody.N.White-at-mcdermott.com
Date: 7/1/97 2:58 PM
Subject: EDX Interpretation

Contents Retrieved from Microscopy Listserver Archives
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I have had some similar samples generate noise in the spectrum which formed
a
peak in the area of carbon's. Usually the low energy side did not return
toward
baseline which was caused me to investigate a bit more. I found that if I
used
my Ultra Thin (metal/polymer) window the unusual response disappeared. If
carbon was really there, the peak would be only slightly attenuated, but
would
head back "down hill" on the low side. I did no more experiments to
determine
the cause, but it was suggested to me by several that the sample was likely
generating photons (~visible spectrum light) which were hitting the crystal
-
causing detector noise...

Regards,
Woody White
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Hello everybody,

Recently we did an SEM/EDX analysis that gave rather baffling results.
After some thought we came up with what seemed to be a plausible
explanation, however, the people for whom the analysis was done remained
skeptical. Therefore I am writing the Listserver for a second opinion.
The sample consisted of irregular shaped alumino-silicate particles roughly
2-5 microns is size. The specimen was uncoated resulting in some charging.
Analysis (windowless) at 1000x mag. resulted in a carbon peak much larger
than the Al or Si peaks. At 5000x the carbon peak was smaller; i.e.,
comparable to the Al and Si. Using a spot mode on individual particles, no
carbon was detected. However, spot mode analysis after lowering the
accelerating voltage from 20 to 10 kv, showed a small carbon peak.
The only explanation we could envision to fit these results was a thin
carbonaceous coating on the particle surface. Is this a plausible
explanation or we missing something? Does anyone have an alternative
suggestion? Any help would be appreciated.

Dan Schwab
Center for Microscopy and Microanalysis
Bowling Green State University
Bowling Green, OH




From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Wed, 2 Jul 1997 14:04:20 -0000
Subject: RE: keeping a protozoa still

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Message-ID: {c=US%a=_%p=Historical_Colle%l=IT3NT-970702140420Z-529-at-it3nt.pasttimes.com}

Thats ok then, as that was how I did tend to pronounce it, I just
thought it may have been pronounced:

PARA-MEK-EE-UM!!

I shall make a note of the useful info on the dyes! I currently have the
standard methyl blue.

Conrad

} -----Original Message-----
} From: Lou Ann Miller [SMTP:lamiller-at-uiuc.edu]
} Sent: Wednesday, July 02, 1997 1:57 PM
} To: Conrad Perfett
} Subject: Re: keeping a protozoa still
}
} Pronouncing paramecium.... the US way:
}
}
} pair a me see um
}
}
}
} I'm not an expert in the field, but I can tell you that ~ 0.5%( grams per
} 100ml) Tolluidine Blue is often used for fungi.
}
} Crystal Violet along with iodine is often used for bacteria that's been
} dried on a slide.
}
} Might try iodine, that would be inexpensive to try, maybe even a generic
} Betadine at the drugstore / pharmacist's.
} Would need a more dilute solution than what form it comes in, so as to not
} overpower your object.
}
}
} Also, speaking as one that has done about 10,000 hospital urine analysis,
} try lowering the condensor by quite a bit, and it makes some things more
} clear at about 40KX objective. ie floating blood cells, mucus strands,
} casts , and a little protozoan called trichamonous. ( forgive my spelling
} , it's been a while, and we just called them trich.) Which you hope you
} never ever have!
}
} Also, 2 pieces of Xray film crossing at right angles ( polorized material)
} will make the background darker, and make things like uric acid crystals
} stand out.
}
}
} Keep having fun!
}
}
}
} Lou Ann
}
}
} } Finally how do you pronounce 'paramecium'?!!!
} }
} } Conrad :)
}
} Lou Ann Miller
} Center for Microscopy & Imaging
} College of Veterinary Medicine
} Dept. of Veterinary Biosciences
} University of Illinois
} Rm 1108 Basic Sciences Bld
} 2001 S Lincoln Ave.
} Urbana, Illinois 61802
}
} Phone: 217-244-1566
} email: lamiller-at-uiuc.edu
}
} Center for Microscopy & Imaging Home page:
} http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
}
} Central States Microscopy Society:
} http://www.cvm.uiuc.edu/Homepages/LouAnnMiller/CSMS/csms.html
}
} Personal Home Pages:
} http://www.cvm.uiuc.edu/Homepages/LouAnnMiller/LAM.html
}
}




From: rtind-at-siu.edu (Randy Tindall)
Date: Wed, 2 Jul 1997 08:00:34 -0600
Subject: EDX Interpretation

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Dan,

How were the samples mounted during analysis? When we do EDX at our lab,
we generally use clean, pure carbon pin-type stubs and carbon adhesive tape
or discs. At a mag of 1000x while viewing particles 2-5 microns in size,
it seems that your probe would be scanning a large area of whatever they
were mounted on. If this is the case, going to higher mag and/or doing
spot analysis, thereby eliminating more of the background, would be likely
to reduce or eliminate the x-rays from the background, and this is what you
observed.

Also, unless I'm mistaken, lowering the accelerating voltage would tend to
emphasize the peaks of lower atomic weight elements, relative to heavier
ones, so if you were getting x-rays from your mounting materials, they
might tend to show up more at lower kv's.

Finally, if you're using a variable pressure scope, lower vacuum in the
column can scatter the electron beam to a large degree. Again, this would
tend to cause x-ray emission from areas other than the particles you are
interested in.

Hope I'm not belaboring the obvious, but it really sounds to me like the
carbon is from your mount.

Randy Tindall
Center for Electron Microscopy
Southern Illinois University at Carbondale






From: Malcolm Thomas :      mgt3-at-msc.cornell.edu
Date: Wed, 2 Jul 1997 09:07:05 -0400 (EDT)
Subject: metallograph recommendations

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Dear microscopists,
We will soon be purchasing an inverted metallograph for specimen prep. It
appears all the major brands are quite good, but perhaps there are subtle
advantages / disadvantages that are only evident after much use and experience.
If anyone has any strong recommendations, I would appreciate your input.
Please feel free to contact me off-line if you prefer. Thanks in advance
for your time and help.

Sincerely,
Mick Thomas

Materials Science Center
Cornell University
Ithaca, NY 14853
mgt3-at-msc.cornell.edu




From: homer-at-biomed.med.yale.edu (R Homer)
Date: Wed, 02 Jul 1997 09:07:22 -0500
Subject: Re: cryo-sectioning

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At 6:59 PM 7/1/97, Linda Barthel wrote:
see
} Barthel & Raymond, J. Histochem Cytochem, 1990, 38:1383-1388 or our web
} site for a detailed protocol, http://www.personal.umich.edu/~praymond/
} Others have used this combination with good success for both thin and
} thick (+10 um) sections.
}
I tried to look up your web site and got an error message to the effect
that the web site could not be found. Is this the right address?

thanks

Rob Homer

Robert Homer, MD, PhD
Asst Prof, Pathology
Yale University School of Medicine
310 Cedar St.
PO Box 208023
New Haven, CT 06520-8023






From: Nestor J. Zaluzec :      Zaluzec-at-Sparc5.Microscopy.Com
Date: Wed, 2 Jul 1997 08:14:34 -0500
Subject: EDS Spectrum Interpretation..

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Just a comment on a comment maninly on semantics...

} the cause, but it was suggested to me by several that the sample was likely
} generating photons (~visible spectrum light) which were hitting the crystal
} causing detector noise...

Visible light does not create noise, it is signal. Remember the SiLi detector
is an ENERGY DISPERSIVE SPECTROMETER. The photons are energy packets just
like X-rays both of which are creating electron/hole pairs. The energy is just
such that is at the very low energy range of the detector and hence in
correctly
gets called noise. It may be signal you do not want to see, but it is
signal.
Preferential attenuation of the light by a very thin visible light
absorbing "window"
is a pretty standard trick to FILTER OUT the light (unwanted signal) from
the low
energy x-rays desired signal.

Nestor
Your Friendly Neighborhood SysOp






From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Wed, 2 Jul 1997 14:07:00 -0000
Subject: RE: keeping a protozoa still

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I guess that is one way! I did think of that, but I guess I wanted to
keep it alive so I can see the processes going on!
Although it may seem obvious, what is the best way to kill them? putting
in something like a few drops of an alcohol solution?

Conrad

} -----Original Message-----
} From: Robert Wieland [SMTP:wieland-at-me.udel.edu]
} Sent: Wednesday, July 02, 1997 1:37 PM
} To: Conrad Perfett
} Subject: Re: keeping a protozoa still
}
} On Wed, 2 Jul 1997, Conrad Perfett wrote:
}
} [lots cut]
} } Is there a method of restricting a protozoa such as paramecium from
} } moving about? The protozoa solution I have is great for studying them,
} } but not if I want to look at the internal structure. Also what stain is
} } a good stain to bring out the details in paramecium such as the nucleus?
}
} There are any number of sources of methyl cellulose, which makes the
} "water" they live in more viscous so they don't dart out of the field. To
} make them absolutely still, I believe it's best to kill them.
}
} }
} } Finally how do you pronounce 'paramecium'?!!!
}
}
} par-ah-ME-see-um
}
} }
} } Conrad :)
} }
} } ------------------------------------------------------------------------
} } -----------
} } "Any sufficiently advanced technology is indistinguishable from magic"
} } ----------------------------------------------------------- Arthur C
} } Clarke ----
} }
} }
}
} Robert Wieland wieland-at-me.udel.edu
} You can't go faster than light, you can't get colder than absolute
} zero, and you can't help somebody by not telling them the truth.
}
}




From: Linda Barthel :      barthel-at-umich.edu
Date: Wed, 2 Jul 1997 09:22:33 -0400 (EDT)
Subject: Re: cryo-sectioning

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Sorry about the web site address, there is one minor error, the correct
address is:
http://www-personal.umich.edu/~praymond/


On Wed, 2 Jul 1997, R Homer wrote:

} At 6:59 PM 7/1/97, Linda Barthel wrote:
} see
} } Barthel & Raymond, J. Histochem Cytochem, 1990, 38:1383-1388 or our web
} } site for a detailed protocol, http://www.personal.umich.edu/~praymond/
} } Others have used this combination with good success for both thin and
} } thick (+10 um) sections.
} }
} I tried to look up your web site and got an error message to the effect
} that the web site could not be found. Is this the right address?
}
} thanks
}
} Rob Homer
}
} Robert Homer, MD, PhD
} Asst Prof, Pathology
} Yale University School of Medicine
} 310 Cedar St.
} PO Box 208023
} New Haven, CT 06520-8023
}
}
}





From: Michael D. Standing :      MDStandi-at-bioag.byu.edu
Date: Wed, 02 Jul 1997 08:29:08 -0700
Subject: EDX Interpretatioin

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Dan,

You don't mention what you mounted your sample on. If it is carbon
based (carbon stub, conductive carbon tape, other adhesive using
cargon), you will see the carbon peak. Going to smaller areas of
excitation (i.e. increasing the relative proportions of sample to
background scanned) will cause the carbon peak to be reduced. At spot
mode on a particle, you are pretty much just exciting the particle and
not the background so you do not see the carbon. At the lower
accelerating voltage the emission volume may broden out closer to the
surface of the background giving you the small carbon peak. It is
important to remember that just because the initial point of excitation
is small, it doesn't mean that the volume of exitation is small. With
your sample, you are probably exciting several cubic microns of your
sample. A good monte carlo program will demonstrate this to you.

Hope this helps

Michael D. Standing
Electron Microscopist
Brigham Young University
e-mail: MDStandi-at-bioag.byu.edu




From: Larry Stoter :      LPS-at-teknesis.demon.co.uk
Date: Wed, 2 Jul 1997 13:33:37 +0100
Subject: Re: Sputter coating with aluminum

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} I have a sample that I have labeled with gold tagged antibody and I want to
} sputter coat it with aluminum for viewing in the SEM. We have a sputter
} coater with a turbomolecular pump and an aluminum target. We tired to
} sputter coat the sample using essentially the same "settings" used for
} chromium, but could not see that any coating had occurred. Does anyone
} have any ideas? Thanks in advance for any help any of you can give this
} humble student.
}
} VTY,
} Beth Bray
} bbray-at-netside.com

Is it possible to successfull sputter Al? Al oxidises rapidly, so I would
have thought you'd have a nice coating of Al oxide, an excellent insulator,
on your specimen.

Perhaps carbon coating would be a better bet?

Regards,
Larry Stoter






From: bozzola-at-siu.edu (John J. Bozzola)
Date: Wed, 2 Jul 1997 09:22:00 -0600
Subject: Re: Sputter coating with aluminum

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} I have a sample that I have labeled with gold tagged antibody and I want to
} sputter coat it with aluminum for viewing in the SEM. We have a sputter
} coater with a turbomolecular pump and an aluminum target. We tired to
} sputter coat the sample using essentially the same "settings" used for
} chromium, but could not see that any coating had occurred. Does anyone
} have any ideas? Thanks in advance for any help any of you can give this
} humble student.

I assume that you are coating with the lower atomic weight Al versus Cr in
order to image the gold using backscattered imaging in the SEM. Lower
atomic numbered metals are difficult to sputter coat properly - although it
can be done using more energy. Also, Al is very reactive and will oxidize
almost immediately after coating. Carbon would provide a better sputter
coat if you have the capability. Actually, thermal evaporation is the
preferred technique with both Al and C.


####################################################################
John J. Bozzola, Ph.D., Director
Center for Electron Microscopy
Neckers Building, Room 146 - B Wing
Southern Illinois University
Carbondale, IL 62901-4402
U.S.A.
Phone: 618-453-3730
Fax: 618-453-2665
Email: bozzola-at-siu.edu
Web: http://www.siu.edu/departments/shops/cem.html
####################################################################






From: Bob_Citron-at-cc.chiron.com (Bob Citron)
Date: 7/1/97 9:25 AM
Subject: TEM preparation of human cornea

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Eugene;

In 1992, I did some SEM studies on corneal endothelial cells from rabbits.
I found that a 0.1M phosphate buffered saline worked fine for that purpose.
Osmolarity was maintained between 300 - 310. This work was published in the
EMSA proceedings for that year; "Evaluation of the Biocompatibility of
Polymer Surface Modifications with the Corneal Endothelium", p. 1106.

Regards,

Bob
***************************
Bob Citron
Chiron Vision
Claremont, CA
USA
(909)399-1311
Bob_Citron-at-cc.chiron.com
***************************


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I am working with the human cornea and wish to find the optimal buffers for
glutaraldehyde and osmium tetroxide dilutions in the preparation of
specimens for TEM. Any suggestions or references would be greatly
appreciated.





From: DANIEL SCHWAB :      DSCHWAB-at-OPIE.BGSU.EDU
Date: Wed, 02 Jul 1997 11:36:38 -0500 (EST)
Subject: EDX Interpretation update

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Hello,

Sorry for the incomplete info in my original posting. Several replies
questioned the mounting method. The specimen was packed into small grooves
in a silica-lead glass -- no carbon containing mountant of any kind was
involved.

Thank you all for your expert advice and interest.

Dan





From: oshel-at-ux1.cso.uiuc.edu (Philip Oshel)
Date: Wed, 2 Jul 1997 11:08:50 -0500
Subject: diamond or other objectives

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How about cubic zirconia for microscope objectives?
RI = 2.15 - 2.45, dispersion = 0.060, birefringence = 0, hardness = 8.5
(quartz = 7),
no cleavage with good toughness.
Diamond is very hard, but has perfect cleavage in 4 directions (which means
it chips relatively easily).
So, CZ gets into diamond's range for RI, and should therefore make high NA
lenses. CZ is a synthetic, so it can be grown under known conditions, and
is relatively cheap and easy to produce. The high dispersion could be a
problem for correcting chromatic aberration, but how much of one?

Phil

} Sic Hoc Legere Scis Nimium Eruditionis Habes {
Philip Oshel
Station A
PO Box 5037
Champaign, IL 61825-5037
(217) 355-1143
oshel-at-ux1.cso.uiuc.edu
*** looking for a job again ******************







From: John Arnott :      ladres-at-worldnet.att.net
Date: Wed, 02 Jul 1997 12:49:24 -0400
Subject: reply: sputter coating with aluminum

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Beth,
Al can be vey differcult to sputter off with DC process since it builds
up a very thick, closely bonded native oxide layer on its surface. You
might want to manually abrade the target to remove the oxide layer, pump
down and sputter immeadiately.
John Arnott




From: Woody.N.White-at-mcdermott.com
Date: 7/2/97 8:11 AM
Subject: EDS Spectrum Interpretation..

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Hello Nestor,

True... I was thinking in terms of undesired signal as noise.
"Undesirable signal" is certainly more accurate!

Speaking of undesirable signal:

Some of my specimens have generated dead times as high as 90+
percent - with the beam OFF! Spectra (w/ beam on) have very
wide peaks and a relatively huge background.
I know the reason... Highly radioactive specimens wreak havoc with
my EDS. Noise - or - signal???? {g}

BTW, I am using an (old) Kevex "Extra" detector. The massive turret
snout does provide a bit of shielding. Would like to add more, but
have no chamber room left (Etec).

Woody

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Just a comment on a comment maninly on semantics...

} the cause, but it was suggested to me by several that the sample was likely

} generating photons (~visible spectrum light) which were hitting the crystal

} causing detector noise...

Visible light does not create noise, it is signal. Remember the SiLi
detector
is an ENERGY DISPERSIVE SPECTROMETER. The photons are energy packets just
like X-rays both of which are creating electron/hole pairs. The energy is
just
such that is at the very low energy range of the detector and hence in
correctly
gets called noise. It may be signal you do not want to see, but it is
signal.
Preferential attenuation of the light by a very thin visible light
absorbing "window"
is a pretty standard trick to FILTER OUT the light (unwanted signal) from
the low
energy x-rays desired signal.

Nestor
Your Friendly Neighborhood SysOp




From: CAMOORE-at-Gems.VCU.EDU
Date: Wed, 02 Jul 1997 13:09:06 -0400 (EDT)
Subject: TEM-collodion coated grids/dna

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HEEEEELP! I am a graduate student trying to get through my LAST
expt and it is not working. I am coating copper grids with
collodion (2% in amyl acetate) and trying to analyze mitochondrial
dna on them. Initially this worked beautifully. The last 200
grids I've done have been a bust. Problems
1)the collodion keeps frying under the beam, it often seems to
"wobble" under the beam. I have tried 3 different bottles of new
collodion, cleaned the grids well with detergent-water-then acetone
rinses all to no avail.

2)the dna seems to have stopped adhering to the collodion. I
first heat nick the dna so it will be open circular (80 degrees, 30
min, in 0.5M NHAc) then I add cytochrome c to bind to the dna (I've
tried concentrations from 2-100ug/ml) I then rotary shadow with
platinum.

Please help, I'm desperate!
Crystal Moore




From: corwinl-at-pt.cyanamid.com
Date: Wed, 02 Jul 1997 13:12 -0500 (EST)
Subject: (P)LM: Advice Wanted on Newer Technologies

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I do polarized light chemical microscopy in the style (or school) of
Walter McCrone as part of doing general analytical chemistry for a
pharmaceutical and chemical manufacturer. Examples of the types of
problems I deal with are relating particle morphology of pure
substances to properties and some rudimentary optical crystallography
to distinguish crystal types. Can anyone provide experience or
references on the possible usefulness of confocal or other "modern"
light microscopic techniques in this area? Are there good journals
covering such topics as well as classical PLM?


Dr. Leonard R. Corwin
Principal Research Chemist
Fort Dodge Animal Health
Cyanamid Agricultural Research Center
Quaker Bridge & Clarksville Roads
PO Box 400
Princeton, NJ 08543-0400
609-716-2278
609-275-5239 fax
corwinl-at-pt.cyanamid.com





From: Linda Fox :      lfox1-at-wpo.it.luc.edu
Date: Wed, 02 Jul 1997 11:57:06 -0500
Subject: LKB knifebreaker, service

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Does anyone know where we can get our LKB 7800B knife breaker
serviced? If possible, by someone in the Chicago area? It is making
sporadically bad knives and we have adjusted all the knobs by the
instruction booklet.
If it needs to be replaced, can anyone recommend a good one? This
one has been with us for over 20 years.

Thanks, Linda Fox, Loyola Univ. Medical Center, Chicago
lfox1-at-wpo.it.luc.edu




From: TRuscica-at-aol.com
Date: Wed, 2 Jul 1997 14:40:24 -0400 (EDT)
Subject: WDX-LVSEM

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To: Didier Le Thiec
I.N.R.A. Centre de Recherches Forestieres
Unite d'Ecophysiologie Forestiere
Laboratoire de Pollution Atmospherique
54280 Champenoux - France

In response to request for information about WDX system with SEM or LVSEM.






From: NANCY SMITH :      nsmith-at-gauss.sci.csuhayward.edu
Date: Mon, 30 Jun 1997 15:03:41 PSD8PDT
Subject: Planaria

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Does anyone have a good SEM fixation method for planaria?

Nancy R. Smith
Director of Operations
Microscope And Graphic Imaging Center
California State University, Hayward
nsmith-at-csuhayward.edu
http://www.csuhayward.edu/SCI/sem




From: shAf :      mshaf-at-darkwing.uoregon.edu
Date: Wed, 02 Jul 1997 12:34:47 -0700
Subject: Re: Sputter coating with aluminum

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John J. Bozzola wrote:

} ...
}
} } I have a sample that I have labeled with gold tagged antibody and I
} want to
} } sputter coat it with aluminum ...
}
} ...
} can be done using more energy. Also, Al is very reactive and will
} oxidize
} almost immediately after coating. ...

An Al oxidation layer tends to be very thin and impervious to further
oxygen ... I might believe that under a vacuum the oxidization wouldn't
occur until exposed to air, and would still be conductive (??)

cheerios, shAf
--
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/}
Michael Shaffer, R.A. - University of Oregon Electron Probe Facility
mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu
http://darkwing.uoregon.edu/~mshaf/






From: Caroline Schooley :      schooley-at-mcn.org
Date: Wed, 2 Jul 1997 12:35:46 -0700
Subject: RE: keeping a protozoa still

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

No, it's MEESE. And for a stain, go to a tropical fish store and get "ich"
medecine. It's an aqueous copper salt & it works fine (see "Fun at 40
Power" in the MMMMMMProject MICRO bibliography)..
Caroline


Caroline Schooley
Educational Outreach Coordinator
Microscopy Society of America
Box 117, 45301 Caspar Point Road
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html
Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/






From: Kirk J C Czymmek :      kirk-at-udel.edu
Date: Wed, 2 Jul 1997 15:45:18 -0400 (EDT)
Subject: RE: keeping a protozoa still

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Dear Conrad,

Try "Protoslo" quieting solution from Carolina Science and Math,
Phone number: 1-800-334-5551.
You might consider requesting a catalog. I believe that you might
find they sell a number of interesting living and dead organisms for
observation by light microscopy.

Regards,

Kirk J. Czymmek, Ph.D.
Biological Electron Microscopy Facility
111 Wolf Hall
University of Delaware
Newark, DE 19716






From: Mike Coviello :      Coviello-at-mae.uta.edu
Date: Wed, 02 Jul 1997 15:22:41 -0500
Subject: Re: Metallographic recommendations

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Dear microscopists,
We will soon be purchasing an inverted metallograph for specimen prep. It
appears all the major brands are quite good, but perhaps there are subtle
advantages / disadvantages that are only evident after much use and
experience.
If anyone has any strong recommendations, I would appreciate your input.
Please feel free to contact me off-line if you prefer. Thanks in advance
for your time and help.

Sincerely,
Mick Thomas

Materials Science Center
Cornell University
Ithaca, NY 14853
mgt3-at-msc.cornell.edu

Dear Mick:

What would you be using this inverted metallographic microscope for?
If you would use it for mostly tripod-type TEM sample preparation, I would
highly recommend Lieca's model. It has a nice design and has higher
resolution than other models that we've tested. If it is for general all
purpose sample preparation and inspection, others would be more qualified
to answer your question.


Regards,


Michael Coviello
EM Lab Manager
The University of Texas -at- Arlington
Arlington, TX
817-272-5496




From: HILDEGARD CROWLEY :      hcrowley-at-du.edu
Date: Wed, 2 Jul 1997 08:39:17 -0600 (MDT)
Subject: Help! Need TEM service contract.

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Hi,

A week before our service contract for our Hitachi-7000 was to be renewed
by Thomas Technical we were informed that they did not have the staffing
for our TEM. We currently are totally without a contract. We cannot
afford Hitachi's offer for the limited service we need. Can anyone
recommend an independent company which we might contact?
Any ideas would be appreciated.
Thanks,
Hildy Crowley
Dept of Biol Sciences
University of Denver
2101 E Wesley Ave
Denver, CO 80208
Tel. 303-871-3026
E-mail { hcrowley-at-DU.edu }
FAX 303-871-3471




From: Frank Karl :      fskarl-at-goodyear.com
Date: Wed, 02 Jul 1997 10:34:09 -0400
Subject: RE: keeping a protozoa still

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} } Subject: Re: keeping a protozoa still

The classic way to quite protozoa was a dilute solution of cocaine HCl.
(Try explaining that to your local DEA officer) A solution of methyl
cellulose will increase the viscosity of the mount to slow activity down
for a good look. Years ago I tried Novocaine, which I obtained from my
dentist, with limited success.

Best wishes...Frank
----------------------------------------------------------------------------
-------

These opinions are mine alone and have no relationship to my employer.
Thank you.

Frank Karl fskarl-at-goodyear.com
Goodyear Tire & Rubber Co. Voice: 330-796-7818
Analytical Services Dept. 415B Fax: 330-796-3304
142 Goodyear Blvd.
Akron, OH 44305
USA

They that give up essential liberty to obtain a little temporary safety
deserve neither liberty or safety. Benjamin Franklin




From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Wed, 2 Jul 1997 10:34:19 -0000
Subject: keeping a protozoa still

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Is there a method of restricting a protozoa such as paramecium from
moving about? The protozoa solution I have is great for studying them,
but not if I want to look at the internal structure. Also what stain is
a good stain to bring out the details in paramecium such as the nucleus?

Finally how do you pronounce 'paramecium'?!!!

We are teaching a summer course to high school kids using protozoans as our subject.
They will be doing a lot of staining to show various internal structures, and a list of these
stains is available on the "under construction" website for the course
(http://131.229.114.77/LAL) - look under the section on examining protozoa. Most of the
solutions are about .01-.05% aqueous. Takes some fussing around to get it right, as
different types of protozoans vary a lot. There is a great text called "Protocols in
Protozoology" edited by Lee and Soldo and published by the Society of Protozoologists. It
has a lot of info on slowing 'em down - ranging from compression under the coverslip to
various exotic techniques. Methylcellulose (10% aqueous, heating (don't boil it) to
dissolve) is good. I have used CO2 (club soda water) added to the drop, as well as .01%
nickel chloride for ciliates. Also try MgCl2 and KCl. Good luck.
Dick Briggs





From: TRuscica
Date: 97-07-02 15:27:54 EDT
Subject: Fwd: WDX-LVSEM

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---------------------
Forwarded message:
Subj: WDX-LVSEM

To: Didier Le Thiec
I.N.R.A. Centre de Recherches Forestieres
Unite d'Ecophysiologie Forestiere
Laboratoire de Pollution Atmospherique
54280 Champenoux - France

In response to request for information about WDX system with SEM or LVSEM.

WDX requires a large probe current which means a large spot size and inferior
resolution. Depending upon the specimen, the large beam current can produce
specimen damage - not a problem for most bulky physical specimens but
probably a problem for organic specimens. Cold stages can reduce damage for
some specimens, but because damage is radiation damage breaking down
molecules anot heat damage, the reduction in damage is not universal.

Operating in the Low Vacuum of Variable Pressure mode does not do anything to
prevent or enhance the beam damage. the residual gas molecules will scatter
the beam, resultiong in x-rays being generated at positions away from the
major beam incidence region. For this reason, care should be exercised in
interpreting the results obtained from x-ray studies in the variable Pressure
Sem mode. Shorter working distances minimize this effect. I hope shortly to
report on another technique for further minimizing this effect.

I can't give you references on WDX in variable pressure SEM, but following
are some references for the early work on the development of the variable
pressure SEM capability.

References on Variable Pressure SEM

V.N.E. Robinson, A Wet Stage Modification to a Scanning Electron Microscope -
Proceedings of the Eighth International Conference on Electron Microscopy,
Ed. J.V. Sanders and D.J. Goodchild, Australian Academy of Science, Canberra,
Vol. II, pp 50-51.

V.N.E. Robinson, A Wet Stage Modification to a Scanning Electron Microscope,
J. Microscopy, 103, pp 71-77, 1975.

V.N.E. Robinson, The Elimination of charging artifacts in the Scanning
Electron Microscope, J. Phys. E: Sci. Instrum. 8, pp 638-640, 1975.

V.N.E. Robinson, Scanning Electron Microscope Environmental Cells, Scanning
Electron Microscopy 1976/I, Ed. O Johari and I Corvin, IITRI, Chicago pp
91-100.

V.N.E. Robinson, The Examination of Hydrated Biological Specimens in a
Scanning Electron Microscope Environmental Cell, Electron Microscopy, 1976,
Proc. 6th European Congress, Ed. Y. Ben-Shaul, Tal International, Jerusalem.
Vol. II, pp 85-90.

D.A. Moncrieff, V.N.E. Robinson and L.B. Harris, Charge Neutralisation of
Insulating Surfaces in the SEM by Gas Ionisation, J. Phys. D: Appl. Phys. Vol
11, pp 2315-2325, 1978.

D.A. Moncrieff, P.R. Barker and V.N.E. Robinson, Electron Scattering by Gas
in the Scanning Electron Microscope, J. Phys. D: Appl. Phys. Vol. 12, pp
481-487, 1979.

G.D. Danilatos and V.N.E. Robinson, Principles of Scanning Electron
Microscopy at High Specimen Chamber Pressures, Scanning, Vol 2, pp 72-82,
1979.

V.N.E. Robinson, the Examination of Hydrated Specimens in electron
Microscopes, in Analysis of Organic Biological Surfaces, John Wiley and Sons,
New York, 1984, Ed. P. Echlin, Ch. 8, pp 191-207.


Tom Ruscica
ETP USA, Electron Detectors Inc.
For V.N.E. Robinson




From: Beth Bray :      bbray-at-netside.com
Date: Wed, 2 Jul 1997 21:47:57 -0400
Subject: Thanks Re: Sputter coating with aluminum

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Message-Id: {199707030154.VAA28308-at-ccs.netside.com}

Thanks to all of you who sent replies in answer to my question about
sputter coating with aluminum. The consensus appears to be that, in
general, "you can't get there from here."
Aluminum was chosen because, as John Bozzola correctly assumed, I want
to image the gold using backscattered imaging in the SEM. I should have
included that little fact in my initial query. My next move will be to
check into using Zn, Cr, Cu, Ag, or Pd--which was another suggestion, and
to research some of the suggested articles, as well as looking at using
thermal evaporation.
There are so many things to learn--whew!--but I am having more fun than
I would ever have thought possible. I am a "mature student" who is
fortunate enough to work for a company that is paying my way back to
college to finish a degree that I started many years ago. I will graduate
in December of this year (God willing and the creek don't rise) and I am
having a blast in school!
Any further ideas will very much welcomed. Again, many thanks!

Beth Bray
bbray-at-netside.com






From: Mike Coviello :      Coviello-at-mae.uta.edu
Date: Wed, 2 Jul 1997 20:57:10 -0500
Subject: Re: Metallographic Recommendations

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Original message:

Dear microscopists,
We will soon be purchasing an inverted metallograph for specimen prep. It
appears all the major brands are quite good, but perhaps there are subtle
advantages / disadvantages that are only evident after much use and
experience.
If anyone has any strong recommendations, I would appreciate your input.
Please feel free to contact me off-line if you prefer. Thanks in advance
for your time and help.

Sincerely,
Mick Thomas

Materials Science Center
Cornell University
Ithaca, NY 14853
mgt3-at-msc.cornell.edu

REPLY:

Dear Mick:

What would you be using this inverted metallographic microscope for?
If you would use it for mostly tripod-type TEM sample preparation, I would
highly recommend Lieca's model. It has a nice design and has higher
resolution than other models that we've tested. If it is for general all
purpose sample preparation and inspection, others would be more qualified
to answer your question.


Regards,


Michael Coviello
EM Lab Manager
The University of Texas -at- Arlington
Arlington, TX
817-272-5496






From: rblyston-at-trinity.edu
Date: Wed, 2 Jul 97 21:36:18 +0100
Subject: RE: keeping a protozoa still part 2

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} The classic way to quite protozoa was a dilute solution of cocaine HCl.

As was mentioned use methyl cellulose. It is sold under the name
Protoslo. Another trick is to chill the Protozoa medium and to use
neutral density filters to keep the heat of the lamp off of the little
buggers.

Blystone in Texas

Robert V. Blystone, Ph.D. {RBLYSTON-at-Trinity.edu}
Professor of Biology
Trinity University
San Antonio, Texas 78212
210.736-7243 210.736-7229 FAX





From: Keith Ryan :      KPR-at-wpo.nerc.ac.uk
Date: Thu, 03 Jul 1997 08:06:34 +0000
Subject: RE: keeping a protozoa still -Reply

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Do I remember tobacco smoke being used for this purpose years ago?

Keith Ryan
Plymouth Marine Lab., UK





From: azriel gorski :      azrielg-at-cc.huji.ac.il
Date: Thu, 3 Jul 1997 11:35:52 +0300 (GMT+0300)
Subject: RE: Toy Microscopes, Resolutions and Aberrations

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Dear Mark,

Sorry to be so long in geting to this. In the rat race of life the rats
are winning. Now to split a few hairs with you Mark, and hopefully we will
not bore the list to death or scare away those amateurs.
On Tue, 1 Jul 1997, Dr. Mark W. Lund wrote:

} } Lenses have several types of aberrations which will cause the loss of
} } detail unless corrected for. Toy microscopes are not.
} }
} } Spherical aberration - "is especially apparent in lenses having sperical
} } surfaces. Light paths near the center of the lens focus at different
} } points compared to light paths near the periphery."
}
}
} The name "spherical aberration" has historical roots in astronomy where
} a spherical mirror has this aberration but a parabolic mirror does not.
} In some instances a spherical mirror will have no spherical aberration
} wheras a parabolic mirror will have maximum spherical aberration.
} The aberration really has nothing to do with the spherical surfaces
} of lenses. The confusion comes because making a surface "aspheric"
} can cure spherical aberration in many instances.

What confusion? Spherical aberation (also sometimes called aperture
aberation) is a recognized term. While the refraction of light remains
constant, the "distance traveled" in the lens and the angles of
the lens to the object change as a function of the curvature of the lens
surface. That curvature resembles the surface of a sphere.

} } } } Field curvature - "is a natural result of using lenses with curved
} } surfaces. The image plane produced by such lenses will be curved. This
} } kind of image occurs in microscopy unless plano (flat-field) objectives
} } are used."
}
} Actually, field curvatureis a natural result of the geometry of the
} real world. Since an object at the edge of the field of view is
} farther from the lens "center" it will tend to be focussed closer to the lens
} than an object on axis. This naturally leads to field curvature unless
} the designer makes the lens weaker for off axis points. It has nothing
} to do with the lenses being curved. The confusion comes from the formula
} for Petzval curvature, which has lens powers in it.

And the geometry you refer to is due to what? I would suggest that it is
due to the curved surface/s of the lens. The curvature of the image does
mimic the curvature of the lens.

As to "off axis points" we are not getting into Coma aberration are we?

To those one the list, hopefully this friendly discussion/hair spliting
session does not bore you all to tears.

Shalom from Jerusalem

Azriel





From: Dr P. Echlin :      pe13-at-cus.cam.ac.uk
Date: Thu, 3 Jul 1997 11:36:42 +0100 (BST)
Subject: Re: Thanks Re: Sputter coating with aluminum

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Linda

as an alternative to having your knife-maker serviced have you considered
replacing the scoring tool part. I am familiar with the LKB 7801A and it's a
simple task to replace the cutting wheel (it should be in the instructions).
If you haven't got any spares then you may have to trace a supplier but a
pack of several replacements would cost you less than the call-out for a
service. My experience with these knife makers is that they virtually go on
for ever.

Sorry if you've already done this.

Malcolm Haswell
e.m. unit
University of Sunderland
UK
----------

Beth:

An additional thought, why not coat with a thin layer of evaporated
carbon. You would only need to put on a few nm.

Patrick Echlin
Cambridge UKOn Wed, 2 Jul 1997, Beth
Bray wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Thanks to all of you who sent replies in answer to my question about
} sputter coating with aluminum. The consensus appears to be that, in
} general, "you can't get there from here."
} Aluminum was chosen because, as John Bozzola correctly assumed, I want
} to image the gold using backscattered imaging in the SEM. I should have
} included that little fact in my initial query. My next move will be to
} check into using Zn, Cr, Cu, Ag, or Pd--which was another suggestion, and
} to research some of the suggested articles, as well as looking at using
} thermal evaporation.
} There are so many things to learn--whew!--but I am having more fun than
} I would ever have thought possible. I am a "mature student" who is
} fortunate enough to work for a company that is paying my way back to
} college to finish a degree that I started many years ago. I will graduate
} in December of this year (God willing and the creek don't rise) and I am
} having a blast in school!
} Any further ideas will very much welcomed. Again, many thanks!
}
} Beth Bray
} bbray-at-netside.com
}
}
}





From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Thu, 3 Jul 1997 13:05:07 -0000
Subject: RE: Amateur Microscopy

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As one fellow amateur to another, I'd be interested in the type of
camera you are intending to use! I currently use a Praktica MTL50 but am
thinking of getting an OM1 or 2. I have looked around and even
secondhand they are still expensive! but it seems Olympus are the only
ones that have the optionable clear screen (which seems to cost anywhere
from 30 to 40 UK pounds for that alone!). I was thinking of following
someones advice on here (from Australia if I remember correctly) and
cementing a square coverslip with canada balsam to the focusing screen
of another cheap camera, probably another Praktica as they are dirt
cheap here secondhand. Anyone else tried this? Is it satisfactory? I
realise it would make the camera only suitable for microphotography but
that does not matter as it would be vastly cheaper.

Conrad

} -----Original Message-----
} From: Frank J. Hogan [SMTP:jhogan-at-freenet.npiec.on.ca]
} Sent: Sunday, June 29, 1997 4:16 AM
} To: Conrad Perfett
} Subject: RE: Amateur Microscopy
}
} Hi again Conrad,
} Been away for a bit and just got caught up with the traffic on the
} mic list....wow! you really started something good. It really is time
} somebody looked into the toy microscope scene or should I say scam. I had
} one when I was a little guy and it really put me off scoping until I got
} my Bushnell lab model for med school.
} I didn't graduate from med school but switched to journalism where I
} earned my bread until recent early retirement. Fortunetely I kept my scope
} after med school days. Now, I hope to make use of it as a hobby tool.
} My scope has all the bells and whistles a lab scope should have I
} guess. I am limited by a three objective turret, 4x,10x,40x and an 8x and
} 10x eyepieces. I am equiped to do photomicroscopy and it is something I'm
} looking forward too.
} From the sound of things all you folks on the list are way ahead of me
} equipment-wise, but I'm content to putt along with what i've got for now.
} It also sounds like the more advanced people are providing all the info
} you can handle.
}
} Very best
} Frank
}




From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Thu, 3 Jul 1997 12:57:00 -0000
Subject: RE: keeping a protozoa still -Reply

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I am a non-smoker! Also I *do* already have a protozoa solution to slow
the critters down, but was looking more for a way to keep them still so
I can look and photograph them

Thanks for the replies I now have a wealth of information!

Conrad :)

} -----Original Message-----
} From: Keith Ryan [SMTP:KPR-at-wpo.nerc.ac.uk]
} Sent: Thursday, July 03, 1997 9:07 AM
} To: fskarl-at-goodyear.com; microscopy-at-Sparc5.Microscopy.Com
} Subject: RE: keeping a protozoa still -Reply
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America




From: admin-at-scottscientific.com (Scott Scientific Inc.)
Date: Thu, 3 Jul 1997 09:28:48 -0400 (EDT)
Subject: Copper Coating

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I am looking for a Canadian facility, perhaps in Quebec that can coat
samples with copper, please email me with any details.


Thank-You in Advance,

Patricia-Ann Harris





From: NICOLA BOCK :      EMZNJB-at-emn1.mateng.nottingham.ac.uk
Date: Thu, 3 Jul 1997 14:33:49 GMT
Subject: Keeping protozoa still

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Conrad,
You could try putting the little critters in the fridge for a while
before you look at them. This works for fruit flies and slows them
down, so it may be worth a try.
Good luck
Nikki

********************************
Nikki Bock
EM Technician
Dept.ME&MD
University Of Nottingham
Nottingham NG7 2RD
Tel: (0115) 9513759
Email: emznjb-at-emn1.nott.ac.uk




From: Rafal Spirydon :      spirydon-at-matlb.kjist.ac.kr
Date: Thu, 3 Jul 1997 22:45:17 +0900
Subject: Simulation of dislocations

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Dear Microscopists

I would have one question. What is software available for simulation of
dislocations?And which is the best?
It may be commercial or non-commercial.

Best regards


Rafal Spirydon





From: Sarka Lhotak :      lhotaks-at-fhs.csu.McMaster.CA
Date: Thu, 3 Jul 1997 10:22:14 -0400 (EDT)
Subject: immuno with anti-BODIPY FL antibody

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I am re-posting my question, since I have not received any replies and
our further trials are still unsuccessfull. Maybe this time, with a
different Subject, somebody will notice and help us. My appologies to
those not interested in immuno.


Dear immuno colleagues,

We would like to label F-actin at the EM level in growth plate
chondrocytes and matrix vesicles. We have done some pilot experiments at
the light microscope level using:

1) Rabbit anti-actin antibody (A 2066 from Sigma) on paraffin sections.
The label was confined only to a subset of smooth muscle cells in control
tissues.

2) Phalloidin-BODIPY FL worked very nicely on cryostat sections,
therefore we tried anti-BODIPY antibody, followed by biotinylated
goat-anti-rabbit, Streptavidin peroxidase and AEC substrate. However, we
got no labelling.

We would appreciate any input on F-actin labelling at EM level in
non-muscle cells or any experience dealing with rabbit anti-BODIPY FL
antibody.

Thank you,

Sarka Lhotak
EM Facility, McMaster University
Hamilton, Ontario, Canada

lhotaks-at-mcmaster.ca






From: Sarka Lhotak :      lhotaks-at-fhs.csu.McMaster.CA
Date: Thu, 3 Jul 1997 10:22:14 -0400 (EDT)
Subject: immuno with anti-BODIPY FL antibody

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I am re-posting my question, since I have not received any replies and
our further trials are still unsuccessfull. Maybe this time, with a
different Subject, somebody will notice and help us. My appologies to
those not interested in immuno.


Dear immuno colleagues,

We would like to label F-actin at the EM level in growth plate
chondrocytes and matrix vesicles. We have done some pilot experiments at
the light microscope level using:

1) Rabbit anti-actin antibody (A 2066 from Sigma) on paraffin sections.
The label was confined only to a subset of smooth muscle cells in control
tissues.

2) Phalloidin-BODIPY FL worked very nicely on cryostat sections,
therefore we tried anti-BODIPY antibody, followed by biotinylated
goat-anti-rabbit, Streptavidin peroxidase and AEC substrate. However, we
got no labelling.

We would appreciate any input on F-actin labelling at EM level in
non-muscle cells or any experience dealing with rabbit anti-BODIPY FL
antibody.

Thank you,

Sarka Lhotak
EM Facility, McMaster University
Hamilton, Ontario, Canada

lhotaks-at-mcmaster.ca






From: Michael D. Standing :      MDStandi-at-bioag.byu.edu
Date: Thu, 03 Jul 1997 08:35:03 -0700
Subject: LKB knifebreaker, service

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Linda,

I believe that Leica is now handeling (or has acquired) the LKB knife
breaker. You may try to contact your local Leica sales rep. for more
information.

Disclaimer: I have no personal or financial interest in Leica

Michael Standing
Electron Microscopist
Brigham Young University
e-mail: MDStandi-at-bioag.byu.edu




From: alexander.black-at-ucg.ie (Alex Black)
Date: Thu, 03 Jul 1997 16:00:23 +0000
Subject: LKB knifebreaker, service

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----------
} From: NANCY SMITH {nsmith-at-gauss.sci.csuhayward.edu}
} To: microscopy-at-Sparc5.Microscopy.Com
} Subject: Planaria
} Date: Mon, 30 Jun 1997 15:03:41 PSD8PDT
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

I think this one works ok, it has been some years since I used it, but it
was fine then!
200ml dist. H2O, 2 ml conc. HNO3, 4.5 ml formalin and 2.5 g MgSO4. Fix by
dropping solution onto the planaria, done at room temp, a higher temp causes
improper relaxation, a lower one, mucus secretion. Fixation complete within
24hrs, the specimens may be stored in 70% ethanol, or 5% formalin. Osmicate
specimens and dehydrate as per usual.
Good luck!

Alex Black
Department of Anatomy
University College Galway
IRELAND






From: Keith Ryan :      KPR-at-wpo.nerc.ac.uk
Date: Thu, 03 Jul 1997 15:48:12 +0000
Subject: Slowing down Protozoa

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Further to my earlier note about tobacco smoke, I now have the
reference: CFA OPantin. Notes on Microscopical Technique for
Zoologists. Cambridge University Press. 1969., page 7.

Funnily enough, guess what it is recommended for - Paramecium !! Also
flagellates, the cilia of Mytilus and Hydra. Not very politically correct (but
in certain matters, I may not be!). Expedient, if not finally terminal for both
the Paramecium and the microscopist!!

Also in this small book (of 76 pages) are recommended:
1. 10% alcohol - if you are an amateur, maybe whisky etc could be
added to your water.
2. magnesium chloride - 7.5% MgCl2.6H2O or 20% MgCl2.7H20 are both
isotonic with seawater, use half seawawater and half MgCl2 solution.
This works atraet within minutes with baby cuttlefish and squid (in fact,
we got a paper about it!)

Use 2.5% Mgcl2.6H2O fo freshwater organisms, it is slow e.g. 2 hours
for Planaria.

3. Menthol. - scatter a few crystals on the surface of the water and
leave overnight. Our specimen dept. used to do this with marine
invertebrates.

4. Ether vapour - good luck (explosively flammable!)

A more modern item is polyethylene oxide with a Molecular Weight of
4,000,000. From the Aldrich catalogue, cat. no. 18,964-4, 5 grams costs
#12.20 in UK before 17.5% tax. Used at 1% in your medium, this works
very well. This is a tip from the person for whom I recently posted a
fluorescence microscopy question.

Best wishes - Keith Ryan
Plymouth Marine Lab., UK (Bonjour, Daniele!)





From: Robert H. Olley :      R.H.Olley-at-reading.ac.uk
Date: Thu, 3 Jul 1997 16:00:47 +0100 (BST)
Subject: OM: Hydrophilic glass coverslips

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http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Does anyone know where we can get hydrophilic glass coverslips, please?
We have seen a reference to them in a German paper, as supplied by
ILMGLAS, but we cannot find this company on the web. Any other suppliers?

Thanks in advance,

+------------------------------------------------------------------------+
| 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: William Barnett - IDL :      barnett-at-amnh.org
Date: Thu, 3 Jul 1997 11:14:53 -0400
Subject: SEM/Confocal Position Available

Contents Retrieved from Microscopy Listserver Archives
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POSITION AVAILABLE: Laboratory Manager, Interdepartmental Laboratories

The American Museum of Natural History is seeking a Laboratory
Manager to manage the Museum's Interdepartmental Laboratories. Duties
include maintenance and operation of analytical microscopy equipment,
particularly a scanning electron microscope with energy dispersive
spectrometer, confocal laser scanning microscope, and digitial imaging and
printing facilities; day to day management of laboratory operations;
training users on analytical instrumentation; and participation in
biological, geological and anthropological research.
Qualifications include an advanced degree in Museum science-related
field, 5 years experience in analytical laboratory operation and with
scanning electron microscopes and energy dispersive systems. Ability to
work with research scientists and to manage day-to-day operation of a
digital imaging facility also required Background in confocal laser
scanning electron microscopy desirable. Full Benefits. Please send resume
with salary history to:

Human Resources
American Museum of Natural History
Central Park West at 79th Street
New York, NY 10024-5192.

You may also e-mail your application to: barnett-at-amnh.org

An Equal Opportunity Employer. No phone calls or faxes, please.

--------------------------------------------------------------------------
| William K. Barnett, Ph.D. |
| Director, Interdepartmental Laboratories barnett-at-amnh.org |
| American Museum of Natural History |
| Central Park West at 79th Street |
| New York, NY 10024-5192 |
--------------------------------------------------------------------------






From: Eric or Pat Metzler :      spruance-at-infinet.com
Date: Thu, 03 Jul 1997 12:34:49 -0400
Subject: inexpensive, high quality microscopes available

Contents Retrieved from Microscopy Listserver Archives
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The recent discussion about inexpensive, high quality microscopes,
reminded me that I have a couple of excess microscopes that I would be
happy to sell. These were my primary instruments until I recently moved
up to Zeiss. They are older, Bausch and Lomb/American Optical Spencer
microscopes with illuminators, excellent objectives, lenses, and
condensors. They are in excellent condition with available lenses
ranging from 3.5x to 100x and include oil immersion. If I remember
right, eyepieces are either 6x or 10x. These are very good for amateur,
exploration, etc. at a low cost. No plastic lenses or cheap parts.

Price: 150 U.S. to $250 U.S. (depending on what you want in the say of
objectives and lenses) plus shipping.

I won't be able to answer you for a few days, but let me know what you
think. I'll get back to you shortly.

Cheers,

Eric

spruance-at-infinet.com




From: JJLIU-at-monsanto.com
Date: Thu, 3 Jul 1997 11:59:37 -0500
Subject: EM Position Available

Contents Retrieved from Microscopy Listserver Archives
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We're redefining the way the world looks at life sciences. We're
the new life sciences company at Monsanto. We're even more
focused on the future. That's why our entire organization, from
agricultural biotechnology to pharmaceuticals to food
ingredients, is dedicated to life sciences. These are areas that
can directly impact our future. Now, we can continuously answer
the questions, needs and demands of our ever-changing planet.
After all, the world's resources are finite, but our vision for
its future is not.


Electron Microscopy Research Associate -

Monsanto Corporate Research has an opportunity for a Research
Associate in electron microscopy. The position is located in the
Analytical Sciences Center (ASC) in St. Louis, MO. The ASC is an
exciting, state-of-the-art research organization serving the
analytical needs of Monsanto's new life sciences company.

The successful candidate will join a small team of experts
involved in the structural examination of catalysts, polymers,
biomaterials, and other nanophase materials. The position
requires an M.S. or B.S. or equivalent in material science or a
related field with 3+ years experience in transmission electron
microscopy (TEM) associated with materials. A strong background
in TEM and associated preparative techniques, including
embedding, ultramicrotomy, and dark room procedures is essential.


The successful candidate will be highly motivated, self-directed,
interested in learning new skills, and effective working
independently or in a team-based research environment. Excellent
interpersonal, verbal and written communication skills are
necessary. Experience with SEM and other imaging techniques, as
well as digital imaging and image analysis are desired but not
required.

Monsanto offers a competitive salary and benefits package, as
well as a challenging and outstanding career opportunity in a
fast-paced scientific organization. Interested applicants should
send their CV or resume (suitable for scanning into our
database), with the names and addresses of three references to:
Monsanto Company, Staffing Code 0276, 800 N. Lindbergh Blvd.,
Mail Zone E2NA, St. Louis, MO 63167. EEO/AA Employer M/F/D/V.
Please visit our website at www.monsanto.com.




From: bozzola-at-siu.edu (John J. Bozzola)
Date: Thu, 3 Jul 1997 12:13:34 -0600
Subject: Re: Thanks Re: Sputter coating with aluminum

Contents Retrieved from Microscopy Listserver Archives
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} Aluminum was chosen because, as John Bozzola correctly assumed, I want
} to image the gold using backscattered imaging in the SEM. I should have
} included that little fact in my initial query. My next move will be to
} check into using Zn, Cr, Cu, Ag, or Pd--which was another suggestion, and
} to research some of the suggested articles, as well as looking at using
} thermal evaporation.

Be careful in chosing the proper metal to coat your specimen because if you
use a metal with too high atomic number (Pd, Ag, Au, Cr, etc) the
backscatter emissions may mask completely the gold emission from the
specimen - especially since it will be weak to begin with. In this case,
thermally evaporated carbon is the coating of choice.

Isn't microscopy fun!
Good luck in your career |8 { ))




####################################################################
John J. Bozzola, Ph.D., Director
Center for Electron Microscopy
Neckers Building, Room 146 - B Wing
Southern Illinois University
Carbondale, IL 62901-4402
U.S.A.
Phone: 618-453-3730
Fax: 618-453-2665
Email: bozzola-at-siu.edu
Web: http://www.siu.edu/departments/shops/cem.html
####################################################################






From: Luc Nocente :      ln-at-noesisvision.com
Date: Thu, 03 Jul 1997 11:07:57 -0400
Subject: Stage controllers

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

I am looking for a stage controller that provides more than 2 microns
accuracy.

Any suggestions?




----------------------------------------------------------------------------
-------------
Luc Nocente Tel: 514 345 1400
Noesis Vision Inc. Fax: 514 345 1575
e-mail: ln-at-noesisvision.com http://www.noesisvision.com
6800 Cote de Liesse, Suite 200
St-Laurent, PQ
H4T 2A7,Canada
----------------------------------------------------------------------------
-------------




From: Juan Marti :      jmartip-at-www.cepade.es
Date: Thu, 03 Jul 1997 20:43:29 +0200
Subject: cathode copper microstructure

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

Hello all,

I have been asked to study copper cathode microstructure with LM. Can
anyone direct me to any descriptions, micrographs or papers written about
cathode copper microstructure. Addresses of Companies and/or organizations
to contact with for help on this subject would also be very helpful to me.

Thanks a lot.

Juan Marti: jmartip-at-cepade.es





From: bozzola-at-siu.edu (John J. Bozzola)
Date: Thu, 3 Jul 1997 14:26:33 -0600
Subject: Re: TEM-collodion coated grids/dna

Contents Retrieved from Microscopy Listserver Archives
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} HEEEEELP! I am a graduate student trying to get through my LAST
} expt and it is not working. I am coating copper grids with
} collodion (2% in amyl acetate) and trying to analyze mitochondrial
} dna on them. Initially this worked beautifully. The last 200
} grids I've done have been a bust. Problems
} 1)the collodion keeps frying under the beam, it often seems to
} "wobble" under the beam. I have tried 3 different bottles of new
} collodion, cleaned the grids well with detergent-water-then acetone
} rinses all to no avail.

Check that you are not over-irradiating the specimen in the TEM (emission
too high, too large condenser spot size or aperture, too low kV). OR you
may have overloaded the grid with specimen material.


} 2)the dna seems to have stopped adhering to the collodion. I
} first heat nick the dna so it will be open circular (80 degrees, 30
} min, in 0.5M NHAc) then I add cytochrome c to bind to the dna (I've
} tried concentrations from 2-100ug/ml) I then rotary shadow with
} platinum.

The cytochrome c may have gone bad. This happened to me once and the dna
was not coated properly. Same batch used as previously?

Is the rotary shadowing being done properly - i.e., adequate Pt being
deposited? If you put down enough Pt, this should stabilize the collodion
as well. Not enough Pt will lead to lack of contrast (no dna seen) as well
as drifting/unstable films.

Did you try checking out plain collodion coated grids, minus specimen? If
bad, try a different vendor.

####################################################################
John J. Bozzola, Ph.D., Director
Center for Electron Microscopy
Neckers Building, Room 146 - B Wing
Southern Illinois University
Carbondale, IL 62901-4402
U.S.A.
Phone: 618-453-3730
Fax: 618-453-2665
Email: bozzola-at-siu.edu
Web: http://www.siu.edu/departments/shops/cem.html
####################################################################






From: Caroline Schooley :      schooley-at-mcn.org
Date: Thu, 3 Jul 1997 16:41:22 -0700
Subject: Re: Slowing down Protozoa

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

An aqueous extract of tobacco leaf usually has enough nicotine to make them
comatose. And if you're a classroom volunteer, it's a dramatic &
definitely PC demonstration for the young folk...
Caroline



Caroline Schooley
Educational Outreach Coordinator
Microscopy Society of America
Box 117, 45301 Caspar Point Road
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html
Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/






From: Kenneth JT Livi :      klivi-at-jhu.edu
Date: Tue, 01 Jul 1997 10:46:45 -0400 (EDT)
Subject: Re: Interfacing TN5500 to PC's and/or networks

Contents Retrieved from Microscopy Listserver Archives
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Hello Ron,

We have a similar setup though we use our TN 5500/5600 on a microprobe.
There is a flex module that enable you to choose to send data out one of
the four ports on the back of your 5500. It is the +} DV command. +} DV 0 is
the default which sends data out the printer port. We send data to +} DV 4
and print out formated information to a PC via a 9-pin cable and into comm
port 1 or 2. This is 1200 baud communications so images take a long time.
On the PC side, we have basic programs that capture what is comming in.
There are programs for both data string (analyses) and image transfer. All
are very limited but being able to transfer data out of the PDP is an
absolute nec.

E-mail if you want more info.

Ciao for now,
Ken

Kenneth JT Livi
Department of Earth and Planetary Sciences
34th and Charles Streets
Johns Hopkins University
Baltimore, Maryland 21218 USA
Phone: (410) 516-8342
Fax: (410) 516-7933
e-mail: klivi-at-jhu.edu






From: Khoo Keng Meng :      medp6023-at-leonis.nus.sg
Date: Fri, 4 Jul 1997 11:19:53 +0800 (SST)
Subject: Re: Slowing down Protozoa

Contents Retrieved from Microscopy Listserver Archives
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Hi!
Just a note on tobacco juice. I was told that stuffing tobacco
leaves on your socks will prevent leaches from crawling up your legs as
you cross rivers. Any scientific truth in it? Anything to do with the
nicotine?

K.M. Khoo

On Thu, 3 Jul 1997, Caroline Schooley wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} } ------------------------------------------------------------------------
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} } -----------------------------------------------------------------------.
} }
} } Further to my earlier note about tobacco smoke, I now have the
} } reference: CFA OPantin. Notes on Microscopical Technique for
} } Zoologists. Cambridge University Press. 1969., page 7.
} }
} } Funnily enough, guess what it is recommended for - Paramecium !! Also
} } flagellates, the cilia of Mytilus and Hydra. Not very politically correct (but
} } in certain matters, I may not be!). Expedient, if not finally terminal for
} } both
} } the Paramecium and the microscopist!!
}
} An aqueous extract of tobacco leaf usually has enough nicotine to make them
} comatose. And if you're a classroom volunteer, it's a dramatic &
} definitely PC demonstration for the young folk...
} Caroline
}
}
}
} Caroline Schooley
} Educational Outreach Coordinator
} Microscopy Society of America
} Box 117, 45301 Caspar Point Road
} Caspar, CA 95420
} Phone/FAX (707)964-9460
} Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html
} Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
}
}
}





From: Stephen Chapman :      PROTRAIN-at-CompuServe.COM
Date: Fri, 4 Jul 1997 03:23:38 -0400
Subject: EDX - Spot Mode

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

I travel the world teaching practical electron microscopy and it worries =
me
the emphasis that people place on "Spot Mode". I do not teach the use of=

spot mode for the following reasons-

1. Just because the spot appears in a certain position on the screen=

is this its correct position. I have found machines many microns out of
step in X and Y directions.

2. Specimens charge and switching out of spot mode does not give the=

operator an easy opportunity to see if the spot had moved during the
analysis.

3. Spot mode gives a false sense of analytical volume, no matter how=

small that spot may be we are almost certainly evaluating microns of
material.

Do others worry about spot mode accuracy, do others test the spot mode
accuracy? Is it not better to simply increase the magnification watching=

the area of interest all the way up before the analysis and all the way
down after the analysis?

What do you think?




From: Heike Buecking :      heibueck-at-uft.uni-bremen.de
Date: Fri, 04 Jul 1997 09:41:38 +0200
Subject: EDX - Spot Mode

Contents Retrieved from Microscopy Listserver Archives
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Dear all,

I have made an experiment with mites and than an embedding with Spurr=B4s
epoxy resin. After polymerisation the blocks are fragile and shatter. Is
there a possiblity to resolve Spurr=B4s and than to repeat the embedding of
the samples or other solutions?

Thanks in advance

Heike Buecking
Dr. Heike Buecking
University of Bremen
UFT
Physiological Plant Anatomy
Leobener Str.
D 28359 Bremen
Germany
TEL: +49-421-218-2954 or
TEL: +49-421-218-7283
FAX: +49-421-218-3737
e-mail: heibueck-at-uft.uni-bremen.de




From: Francois Goutenoire :      francois-at-fluo.univ-lemans.fr
Date: Sat, 05 Jul 1997 09:52:39 +0100
Subject: unsubscribe

Contents Retrieved from Microscopy Listserver Archives
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unsubscribe
=================================================================Dr.
Francois Goutenoire
Laboratoire des Fluorures - UPRES A 6010
Faculte des Sciences-Universite du Maine
Avenuue O. Messian, BP 535 Fax:(33).2.43.83.35.06 72000 Le
Mans Cedex, France Tel:(33).2.43.83.33.53
=================================================================


Phone:(0033).2.43.83.33.53
E-Mail: francois-at-fluo.univ-lemans.fr





From: Heike Buecking :      heibueck-at-uft.uni-bremen.de
Date: Fri, 04 Jul 1997 10:01:00 +0200
Subject: =?iso-8859-1?Q?Problems_with_Spurr=B4s?=

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

} Date: Fri, 04 Jul 1997 09:41:38 +0200
} To: microscopy-at-msa.microscopy.com
} From: Heike Buecking {heibueck-at-uft.uni-bremen.de}
}
} Dear all,
}
} I have made an experiment with mites and than an embedding with Spurr=B4s
epoxy resin. After polymerisation the blocks are fragile and shatter. Is
there a possiblity to resolve Spurr=B4s and than to repeat the embedding of
the samples or other solutions?
}
} Thanks in advance
}
} Heike Buecking
Dr. Heike Buecking
University of Bremen
UFT
Physiological Plant Anatomy
Leobener Str.
D 28359 Bremen
Germany
TEL: +49-421-218-2954 or
TEL: +49-421-218-7283
FAX: +49-421-218-3737
e-mail: heibueck-at-uft.uni-bremen.de




From: ainswort-at-geology.gla.ac.uk (Pete Ainsworth)
Date: Fri, 4 Jul 1997 12:38:21 +0100
Subject: Re: EDX - Spot Mode

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

I would agree with Stephen that it is important to emphasise the possible
inaccuracies with using spot mode. However I would not go as far as saying
don't use spot mode.

Spot mode can be useful for both quick looks to check compositions of a
variety of particles in a field of view as well as for accurate
microanalysis. Obviously with the latter it is important to be aware of the
possibility of drift and to be sure of the position of the spot. Certainly
with thin sections it is often possible to see the burn marks where the
beam hit the sample. I tend to emphasise to students that when using spot
mode the analysis will be a volume of 1-2 microns diameter and depth
depending on sample and conditions. Even when using general rastering the
same problem will be there.

This is just my opinion.


} I travel the world teaching practical electron microscopy and it worries me
} the emphasis that people place on "Spot Mode". I do not teach the use of
} spot mode for the following reasons-
}
} 1. Just because the spot appears in a certain position on the screen
} is this its correct position. I have found machines many microns out of
} step in X and Y directions.
}
} 2. Specimens charge and switching out of spot mode does not give the
} operator an easy opportunity to see if the spot had moved during the
} analysis.
}
} 3. Spot mode gives a false sense of analytical volume, no matter how
} small that spot may be we are almost certainly evaluating microns of
} material.
}
} Do others worry about spot mode accuracy, do others test the spot mode
} accuracy? Is it not better to simply increase the magnification watching
} the area of interest all the way up before the analysis and all the way
} down after the analysis?
}
} What do you think?

**************************************
* Pete Ainsworth *
* Dept. Geology & Applied Geology *
* Lillybank Gardens *
* University of Glasgow *
* Glasgow G12 8QQ *
* e-mail: ainswort-at-geology.gla.ac.uk *
* Tel : 0141 330 5505 (direct) *
**************************************






From: Bill Miller :      microbill-at-MOHAWK.NET
Date: Fri, 04 Jul 1997 09:33:38 -0400
Subject: Re: EDX - Spot Mode

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

Stephen Chapman wrote:

} ------------------------------------------------------------------------
}
} The Microscopy ListServer -- Sponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- Send Email to
} ListServer-at-MSA.Microscopy.Com
} -----------------------------
} -----------------------------------------.
}
} I travel the world teaching practical electron microscopy and it
} worries me
} the emphasis that people place on "Spot Mode". I do not teach the use
} of
} spot mode for the following reasons-
}
} 1. Just because the spot appears in a certain position on the
} screen
} is this its correct position. I have found machines many microns out
} of
} step in X and Y directions.
}
} 2. Specimens charge and switching out of spot mode does not give
} the
} operator an easy opportunity to see if the spot had moved during the
} analysis.
}
} 3. Spot mode gives a false sense of analytical volume, no matter
} how
} small that spot may be we are almost certainly evaluating microns of
} material.
}
} Do others worry about spot mode accuracy, do others test the spot mode
}
} accuracy? Is it not better to simply increase the magnification
} watching
} the area of interest all the way up before the analysis and all the
} way
} down after the analysis?
}
} What do you think?

Stephen - you are absolutely right - infact the only way to be sure
where the spot was is to look for the specimen damage after the fact! If
you have to analyze a small area (first think about the excitation
volume) you are better off doing a redcued area scan or go to high
magnification so you can at least "see" where the beam is hitting the
sample.

Bill Miller





From: Sara Prins :      SPrins-at-csir.co.za
Date: Fri, 04 Jul 1997 16:48:56 +0300
Subject: Electron diffraction patterns

Contents Retrieved from Microscopy Listserver Archives
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I am looking for an electron diffraction analysis software package. I have
to identify some particles (wether they're cubic, tetragonal, etc) and
there must be an easier way than to hand draw the expected diffraction
patterns....Does anybody know of something useful on the Web?

Thanx
Sara

==========================================================
Sara Prins
Surface and Structure Analytical Services
Division for Material Science and Technology
CSIR
PO Box 395
Pretoria
SOUTH AFRICA




From: Dr. Andrew P. Somlyo :      aps2n-at-elvis.med.virginia.edu
Date: Fri, 4 Jul 1997 11:13:03 -0400 (EDT)
Subject: Re: EDX - Spot Mode

Contents Retrieved from Microscopy Listserver Archives
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Depending on the stability of your stage/specimen, you should verify
the position of the spot at required intervals. My collegues and I and
numerous grad. students and post-docs have used this approach for decades.



On Fri, 4 Jul 1997, Bill Miller wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Stephen Chapman wrote:
}
} } ------------------------------------------------------------------------
} }
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of
} } America
} } To Subscribe/Unsubscribe -- Send Email to
} } ListServer-at-MSA.Microscopy.Com
} } -----------------------------
} } -----------------------------------------.
} }
} } I travel the world teaching practical electron microscopy and it
} } worries me
} } the emphasis that people place on "Spot Mode". I do not teach the use
} } of
} } spot mode for the following reasons-
} }
} } 1. Just because the spot appears in a certain position on the
} } screen
} } is this its correct position. I have found machines many microns out
} } of
} } step in X and Y directions.
} }
} } 2. Specimens charge and switching out of spot mode does not give
} } the
} } operator an easy opportunity to see if the spot had moved during the
} } analysis.
} }
} } 3. Spot mode gives a false sense of analytical volume, no matter
} } how
} } small that spot may be we are almost certainly evaluating microns of
} } material.
} }
} } Do others worry about spot mode accuracy, do others test the spot mode
} }
} } accuracy? Is it not better to simply increase the magnification
} } watching
} } the area of interest all the way up before the analysis and all the
} } way
} } down after the analysis?
} }
} } What do you think?
}
} Stephen - you are absolutely right - infact the only way to be sure
} where the spot was is to look for the specimen damage after the fact! If
} you have to analyze a small area (first think about the excitation
} volume) you are better off doing a redcued area scan or go to high
} magnification so you can at least "see" where the beam is hitting the
} sample.
}
} Bill Miller
}
}




From: bozzola-at-siu.edu (John J. Bozzola)
Date: Fri, 4 Jul 1997 10:55:04 -0500
Subject: RE:Spurrs embedding problem

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

} I have made an experiment with mites and than an embedding with Spurr=B4s
} epoxy resin. After polymerisation the blocks are fragile and shatter. Is
} there a possiblity to resolve Spurr=B4s and than to repeat the embedding of
} the samples or other solutions?

Spurr's is notoriously susceptible to moisture which, when present,
results in a brittle block that shatters during trimming. The moisture may
be absorbed from the atmosphere if one allows the uncapped specimens to sit
overnight (presumably to evaporate propylene oxide) or, of course, it may
not have been removed properly from the specimen during the dehydration
stage. I suspect that your resin has absorbed moisture from the atmoshere.
Check this out by polymerizing a block of plain resin. If it shatters, toss
out any components that may have absorbed moisture and start over. Spurr's
is a good resin but it is toxic, potentially carcinogenic and sometimes a
pain. Take precautions against contact (nitrile gloves), inhalation (fume
or exhaust hood) and properly dispose of the components by polymerizing
them.


###########################
Dr. John Bozzola, Director
Center for Electron Microscopy
Southern Illinois University
Carbondale, IL 62901
Phone: 618-453-3730
=46ax: 618-453-2665
###########################






From: SSARDARI-at-pharmacy.ualberta.ca
Date: Fri, 4 Jul 1997 12:15:19 MDT
Subject: Yeast cell TEM

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Dear friends,
I have some problem in interpreting my transmittance electron
microscopy. Please, let me know if you can help me.
Sincerely
Soroush Sardari
PhD student,
Faculty of Pharmacy,
Univ. of Alberta,
Edmonton, Canada, T6G 2N8
Fax: (403) 492-1217




From: Nestor J. Zaluzec :      Zaluzec-at-Sparc5.Microscopy.Com
Date: Sat, 5 Jul 1997 12:01:29 -0500
Subject: June Archives On-Line

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Colleagues

The June 97 Microscopy Listserver Archives are
now on-line. You may find them on the MSA WWW
site under Education/Reference Section.

http://www.msa.microscopy.com


Nestor
Your Friendly Neighborhood SysOp






From: Warren Straszheim :      wes-at-ameslab.gov
Date: Sat, 05 Jul 1997 14:43:44 -0500
Subject: Re: EDX - Spot Mode

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I routinely demonstrate to users of our JEOL 840 that they need to be
careful of hysteris during scanning. We have TV rate, a rapid scan, and two
slow scan modes on our SEM. I switch on the cross hair display at TV rate
and select a point of interest. I then switch to rapid scan and have to
recenter the cross hairs. I then switch to slow scan mode and recenter
again. Then I ask them which is correct? Of course the slower scan is most
nearly correct. I can demonstrate by watching the brightness LEDs as I move
the spot across a bright feature.

Fortunately, we are not using spot mode, per se, all that much. Both our
scopes now have digital imaging with point-and-shoot x-ray analysis. It is
much easier locating the point(s) of interest. But users still need to be
aware that there may some lag in the scan while recording the digital image.
Since we tend to use rather slow scans that is not much of a problem. But
users still need to be aware.

Depending on the rush, I still often use your method of cranking up the
manification so that the feature fills the field of view. I am fairly
assured that what I see is what I get for x-rays.

At 03:23 AM 7/4/97 -0400, you wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

E-Mail: wes-at-ameslab.gov (or: wesaia-at-iastate.edu)
http://www.public.iastate.edu/~iprt_info/cfce/ (re: coal)
http://www.public.iastate.edu/~wesaia/marl/ (re: SEM)

electron microscopy, x-ray analysis, image analysis
computer applications
coal characterization and processing





From: Phil Fraundorf :      philf-at-NEWTON.UMSL.EDU
Date: Sun, 06 Jul 1997 01:43:48
Subject: Viewing your scope from all angles

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Hi,

One way to record experimental data on
objects, rooms, and even reciprocal-space,
may be to record images along directions
evenly-distributed over the surface of a
sphere. As Bucky Fuller pointed out in our
neck of the woods some years ago, the
20 face-normals of an icosahedron
are especially convenient in this regard,
in part because the pictures required can be
shot with a single 24-exposure roll of film.

To illustrate, HTML templates for exploring
and animating data-sets on a Mathematica-drawn
Klein bottle, and our Philips EM430ST in its
new low-vibration site, are now on the web*.
A technical note on the angles for taking such
pictures is linked to each template set. Let
me know if you decide to make available similar
views from your own labs, or have other
application-related questions we might help
with.

Enjoy! /philf :)

* http://www.umsl.edu/~fraundor/3d20/index.html



\\/
(-at- -at-)
//\/\/\/\--o0O-(_)-Ooo--}
//P.Fraundorf Phys&Astr/CME 3145165044 philf-at-newton.umsl.edu
\\U.Missouri-St.Louis MO 63121 http://www.umsl.edu/~fraundor
\\/\/\/\/\/\/\/\/----------------}




From: rick-at-pgt.com (Rick Mott)
Date: Mon, 7 Jul 97 10:03:12 EDT
Subject: Re: EDX - Spot Mode

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shAf wrote:

} A major problems with scanning at high mag are either the beam is
} blanked while the beam waits for "start frame" or "start line" sync
} signals, thus count times are inaccurate ... or, if the beam isn't
} blanked then the volume analyzed is weighted to the upper left corner
} and left edge ...

These aren't the only alternatives. I can't speak for other EDS
manufacturers, but our system links the scan generator and the X-ray
pulse processor so that raster retrace and settling times are treated
as "dead time". So the counting (live) time is correct and the pixel
weightings are equal.

Regards to all,

Rick Mott, PGT
rick-at-pgt.com
www.pgt.com




From: William Tivol :      tivol-at-wadsworth.org
Date: Mon, 7 Jul 1997 11:15:44 -0500 (EDT)
Subject: Re: EDX - Spot Mode

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} 1. Just because the spot appears in a certain position on the screen
} is this its correct position. I have found machines many microns out of
} step in X and Y directions.

In my case, I'm always worried about this. I have, however, found
that for every electron-opaque object I have measured I have found the ap-
propriate element(s) when the beam is on the object and not when it is
~1 micrometer away. In many cases, there is a dark spot at the measured
site after the measurement, but not before. Since these spots are at the
position where the beam was set, and since occasional specimen drifts are
accompanied be shifts of the spot and lowered or absent peaks (compared
to subsequent measurements), I think I can be confident about the positions
of my analyses.
}
} 2. Specimens charge and switching out of spot mode does not give the
} operator an easy opportunity to see if the spot had moved during the
} analysis.
}
A distinct advantage of the use of high voltage (~1MV) and very low
beam current is that charging is very seldom a problem. Since I don't have
scanning capability, it is of little concern if I have to take a long time
to accumulate counts. If I were to do mapping, however, low beam current
would be very detrimental.

} 3. Spot mode gives a false sense of analytical volume, no matter how
} small that spot may be we are almost certainly evaluating microns of
} material.
}
Especially in my case where the typical specimen is a biological
section ~1 micrometer thick. David Joy's Monte Carlo program shows pretty
well what the measurement volumes are.

} Do others worry about spot mode accuracy, do others test the spot mode
} accuracy? Is it not better to simply increase the magnification watching
} the area of interest all the way up before the analysis and all the way
} down after the analysis?

Having complete control of the spot size by being able to set the
condenser aperture and two condenser lenses independently of the magnifica-
tion, I just use standard settings for each analysis--10K mag, 30 micrometer
C2 aperture, maximally excited C1 lens, C2 to crossover.
Yours,
Bill Tivol




From: yuhui xu :      Yuhui=Xu%RES%DFCI-at-EYE.DFCI.HARVARD.EDU
Date: Mon, 7 Jul 97 11:21:07 EDT
Subject: Help on SEM sample preparation

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Dear Colleagues:
Recently we came across a sample for SEM which we have great difficulty to
process. We would like to ask for your help:
This is a kind of poly-electrolyte polymer, that has been treated to form
microspheres ( 1 to 10 micron in diameter ) and cross-linked to maintein the
folded shape. Since the beads are highly negatively charged they do not stick
to poly-L-lysine coated cover slips. We tried to run the pellets in Eppendorf
tubes through alcohol dehydration, critical point drying, and then sprinkled
the dried ( as powder) onto double-sided scotch tape and sputter coated. The
results are generally not satisfactory partly due to the fact that it is very
difficult to dry these beads thoroughly in tubes. I wonder whether there have
been better methods for this type of sample which we do not know of. Any
suggestions will be very much appreciated.
Best regards,
Yuhui Xu




From: Larry Stoter :      LPS-at-teknesis.demon.co.uk
Date: Sun, 6 Jul 1997 16:23:07 +0100
Subject: Re: EDX - Spot Mode

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} I travel the world teaching practical electron microscopy and it worries me
} the emphasis that people place on "Spot Mode". I do not teach the use of
} spot mode for the following reasons-
}
} 1. Just because the spot appears in a certain position on the screen
} is this its correct position. I have found machines many microns out of
} step in X and Y directions.
}
} 2. Specimens charge and switching out of spot mode does not give the
} operator an easy opportunity to see if the spot had moved during the
} analysis.
}
} 3. Spot mode gives a false sense of analytical volume, no matter how
} small that spot may be we are almost certainly evaluating microns of
} material.
}
} Do others worry about spot mode accuracy, do others test the spot mode
} accuracy? Is it not better to simply increase the magnification watching
} the area of interest all the way up before the analysis and all the way
} down after the analysis?
}
} What do you think?

While the spot mode leaves a lot to be desired, particularly if the user
isn't aware of the problems, analysis in raster mode is not without its
difficulties. Most SEMs use some sort of of sychronisation of the line and
frame scan signals. If so, the analysis obtained in raster mode is biased
towards one edge and one corner of an area which may not even be identical
with the image area - the beam 'waits' at the beginning of each line and
each frame to synch, and the display may actually be blanked for a small
distance after the scan starts and before the scan ends (over scanning) to
hide image distortions arising from hysteresis, which is usually most
evident at the edges of the scans.

Get a nice dirty specimen and scan it with a coarse raster until the
contamination builds up. Now image the area at a lower mag, and look at the
contamination pattern - there will probably be a heavier contamination line
down one edge (the line scan 'wait') and a spot at one corner (the frame
'wait'). In addition, you may also be unlucky enough to find that the lines
making up the coarse raster are bent at each end - you don't see the
distortion in your images however because this part of the frame is blanked
from display, but it will contribute to an EDX analysis.

More modern SEMs have reduced these problems, but I believe that they are
still present in many.

I think that this demonstrates that whatever 'employers' might want, and
manufacturers try to provide, for anything beyond the most basic EM, you
need a skilled and trained operator with a full understanding of the
instrumentation, and the time to fully check and calibrate the machine (who
needs to be properly paid for undertaking a highly sophisticated and
skilled job). Otherwise you get results that are at best doubtful and at
worst wrong.

Regards,
Larry Stoter






From: Gary Liechty :      garyliechty-at-worldnet.att.net
Date: Mon, 07 Jul 1997 10:26:42 -0700
Subject: Re: metallograph recommendations

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Malcolm Thomas wrote:
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Dear microscopists,
} We will soon be purchasing an inverted metallograph for specimen prep. It
} appears all the major brands are quite good, but perhaps there are subtle
} advantages / disadvantages that are only evident after much use and experience.
} If anyone has any strong recommendations, I would appreciate your input.
} Please feel free to contact me off-line if you prefer. Thanks in advance
} for your time and help.
}
} Sincerely,
} Mick Thomas
}
} Materials Science Center
} Cornell University
} Ithaca, NY 14853
} mgt3-at-msc.cornell.edu
Hi Mick,

Several of my customers have the new Leica. It is a low profile scope,
provides easy access, very little stage drift and great amount of
working distance. It is a new design and seems to be very popular.
Call the 800 number for information for their number (800) 555-1212.

With regards to my recommendation for the LocTite 460, how did it
perform?

Good Luck,

Sincerely,

Gary Liechty
Allied High Tech Products
2376 E. Pacifica Place
Rancho Dominguez, Ca 90220

800-675-1118
310-762-6808 Fax

Products for Materiallographic, SEM and TEM Sample Preparation




From: King :      King-at-bioscience.biology.utah.edu
Date: 7 Jul 1997 12:15:50 U
Subject: Diamond Knives

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

I will soon need to get one or more new diamond knives for use with
biological specimens. I haven't purchased a new knife for several years, and
am curious about the quality of knives currently available. I'd appreciate
hearing from anyone who has purchased and used a new knife in the last year
or so.

Thank you.

Ed King
king-at-bioscience.utah.edu







From: Linda Barthel :      barthel-at-umich.edu
Date: Mon, 7 Jul 1997 16:06:41 -0400 (EDT)
Subject: Glycogen

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Does anyone know of a method/stain that would localize glycogen in cryo
sections of tissue (specifically goldfish neural retina) fixed with 4%
paraformaldehyde? We are interested in the light level, not EM.
Linda Barthel
Research Associate II
Department of Anatomy and Cell Biology
University of Michigan
lab (313) 764-7476
fax (313) 763-1166
barthel-at-umich.edu






From: Ming Chen :      mingchen-at-gpu.srv.ualberta.ca
Date: Mon, 7 Jul 1997 14:38:49 -0600 (MDT)
Subject: Re: Help on SEM sample preparation

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Hi yuhui,

You may simply put one drop of your sample onto a 0.45 u milipore filter
or a normal stub and air dry it for 2-3 hours.

Good luck,


On Mon, 7 Jul 1997, yuhui xu wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Dear Colleagues:
} Recently we came across a sample for SEM which we have great difficulty to
} process. We would like to ask for your help:
} This is a kind of poly-electrolyte polymer, that has been treated to form
} microspheres ( 1 to 10 micron in diameter ) and cross-linked to maintein the
} folded shape. Since the beads are highly negatively charged they do not stick
} to poly-L-lysine coated cover slips. We tried to run the pellets in Eppendorf
} tubes through alcohol dehydration, critical point drying, and then sprinkled
} the dried ( as powder) onto double-sided scotch tape and sputter coated. The
} results are generally not satisfactory partly due to the fact that it is very
} difficult to dry these beads thoroughly in tubes. I wonder whether there have
} been better methods for this type of sample which we do not know of. Any
} suggestions will be very much appreciated.
} Best regards,
} Yuhui Xu
}


***********************************************
* Ming H. Chen, PhD *
* Medicine/Dentistry Electron Microscopy Unit *
* University Of Alberta. *
* Edmonton, Alberta, Canada *
* *
* Visit My Page At: *
* http://www.ualberta.ca/~mingchen *
***********************************************








From: gllovel-at-ppco.com (Gary Lovell)
Date: Mon, 7 Jul 1997 16:15:17 -0500
Subject: ESEM EDS ?

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I have been trying to identify ESEM EDS resolution using a 50 micron
diameter Nickel standard embedded in epoxy (C, O and Cl). My instrument
parameters are 1: 15kv 2: 12.0 mm working distance 3: Chamber
pressure=3.5T 4: Chamber Gas = N2 or H2O (did'nt make any difference which
gas was used) 5: Condensor setting = 50% 6: Sample tilt = 30, 20, 10.
Doing a spot analysis in the center of the Ni standard showed abundant C and
O, and some Cl as well as the expected Ni counts. Is the skirting effect of
the ESEM such that one can't perform EDS on smples of 50 microns or less? I
would be interested in hearing from anyone who has any results for ESEM EDS
resolution.





From: Geoff McAuliffe :      mcauliff-at-UMDNJ.EDU
Date: Mon, 07 Jul 1997 16:40:09 -0700
Subject: Re;glycogen

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Linda:

PAS will stain glycogen but also other things in frozen sections. Can
the sections be treated to reomve lipids? Otherwise, how about Best's
Carmine? Give a day or two and I'll look in Lillie and Fullmer.

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 e-mail: mcauliff-at-umdnj.edu
***************************************************************




From: Stuart McKernan :      mckernan-at-cems.umn.edu
Date: Mon, 7 Jul 1997 19:04:15 -0500
Subject: Re: ESEM EDS ?

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Responding to the message of {199707072115.AA24968-at-relay.ppco.com}
from gllovel-at-ppco.com (Gary Lovell):
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} I have been trying to identify ESEM EDS resolution using a 50 micron
} diameter Nickel standard embedded in epoxy (C, O and Cl). My instrument
} parameters are 1: 15kv 2: 12.0 mm working distance 3: Chamber
} pressure=3.5T 4: Chamber Gas = N2 or H2O (did'nt make any difference which
} gas was used) 5: Condensor setting = 50% 6: Sample tilt = 30, 20, 10.
} Doing a spot analysis in the center of the Ni standard showed abundant C and
} O, and some Cl as well as the expected Ni counts. Is the skirting effect of
} the ESEM such that one can't perform EDS on smples of 50 microns or less? I
} would be interested in hearing from anyone who has any results for ESEM EDS
} resolution.
}
}
From the literature on this subject I think that the chamber pressure is too
high; resulting in too much scattering of the incoming beam. There are several
papers by Brendon Griffin in Australia, and by the Danish group in RISO -
Horsewell, Appel and Bilde-Sorensen - where they have extensively documented the
EDS resolution as a function of voltage, pressure and working distance. See MSA
proceedings from 1995 and 1996 for papers from the ESEM symposia which cover
this topic

Good luck,



__________________
Stuart McKernan stuartm-at-tc.umn.edu
Microscopy Specialist
CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594
100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590





From: Glen MacDonald :      glenmac-at-u.washington.edu
Date: Mon, 07 Jul 1997 17:26:33 -0800
Subject: Re: Slowing down Protozoa

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In my undergrad. zoology lab long ago we added 7-Up to slow down marine
invertebrates. Essentially they are a bit oxygen starved from all the
CO2. Nicotine would cause larger organisms, such as chitons, to curl
and distort.

On Thu, 3 Jul 1997 16:41:22 -0700 schooley-at-mcn.org (Caroline Schooley)
wrote:

} }
} } Further to my earlier note about tobacco smoke, I now have the
} } reference: CFA OPantin. Notes on Microscopical Technique for
} } Zoologists. Cambridge University Press. 1969., page 7.
} }
} } Funnily enough, guess what it is recommended for - Paramecium !! Also
} } flagellates, the cilia of Mytilus and Hydra. Not very politically
} correct (but
} } in certain matters, I may not be!). Expedient, if not finally terminal
} for
} } both
} } the Paramecium and the microscopist!!
}
} An aqueous extract of tobacco leaf usually has enough nicotine to make
} them
} comatose. And if you're a classroom volunteer, it's a dramatic &
} definitely PC demonstration for the young folk...
} Caroline
}
}
Glen MacDonald
Virginia Bloedel Hearing Research Center
Box 35-7923
University of Washington
Seattle, WA 98195-7923
(206) 616-4156
glenmac-at-u.washington.edu
*---------------------------------------------------------------------*
The box said "Requires Windows 95 or better.", so I bought a Macintosh.
*---------------------------------------------------------------------*





From: Ciprian Almonte :      calmonte-at-pitt.edu
Date: Mon, 7 Jul 1997 22:35:23 -0500
Subject: High Resolution Mosaic

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Hi Guys,
I'm currently acquiring mosaic images of muscle fibers with Mosaic Tiling
under Oncor Image version 2.0.5d. However, the copy of Mosaic Tiling that
I have doesn't allow me to get high resolution mosaic image. Does anyone
know if there is a copy of Mosaic Tiling for Oncor or anyother alternative
that will allow me to get high resolution mosaic images good for
quantification.
Thanks,


--Ciprian
Have fun and keep the sun on your back and a smile on your face.
__________________________________________________________
Ciprian A. Almonte
University of Pittsburgh
Center for Biologic Imaging
Pittsburgh, PA 15261

Visit my web site at http://www.pitt.edu/~calmonte
Laboratory's website: http://sbic6.sbic.pitt.edu
__________________________________________________________






From: Mary Mager :      mager-at-unixg.ubc.ca
Date: Mon, 07 Jul 1997 19:35:56 -0700
Subject: Re: Metalergical microscopes

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Dear Roadwalk,
The main difference between a biological and metallurgical light microscope
is that the metallurgical microscope views in the reflection mode, where the
light comes through the objective lens and reflects off the opaque sample,
since very few metal samples are transparent to light. Many metallurgical
microscopes are inverted, that is, the sample is put upside down on a stage
on the top of the microscope, with the objective nosepiece underneath, so
there is very little restriction on sample size. Several of our microscopes
are dual purpose and can work in either transmitted or reflected light mode.
They look like an ordinary biological microscope.

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


Regards,
Mary
Mary Mager
Electron Microscopist
Metals and Materials Eng., UBC
6350 Stores Rd.
Vancouver, B.C. V6T 1Z4
CANADA
tel:604-822-5648, fax:604-822-3619
e-mail: mager-at-unixg.ubc.ca





From: POSTMASTER-at-msmail.his.tch.tmc.edu
Date: Mon, 7 Jul 1997 07:21:00 -0500
Subject: Mail failure

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

Dear Sir,

We would like to examine a pore size of cellulose acetate with SEM. Please
advise us, how to prepare the sample becuase it is very sensitive to
vacuum. Can we dry by using a CPD?

Thank you

Paiboon







From: I.Montgomery-at-bio.gla.ac.uk (Ian Montgomery)
Date: Tue, 8 Jul 1997 08:57:53 +0100
Subject: Re: Glycogen

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Linda,
Periodic Acid Schiff is first favourite. Any histochemistry
text will give you the necessary details, failing that, e-mail me and I can
send you my protocol.
Ian.






From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Tue, 8 Jul 1997 08:56:59 -0000
Subject: Re: Glycogen

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well I now know why I am having difficulty finding the Amoeba since
according to the book I have just bought, at least the classic Amoeba
Proteus is rare in the wild!!! I remember being told by a retired
microscopist that it was rare also. So at least I can take comfort in
the fact that its more than likely there are none in my samples rather
than my mis-identification! But I will not be giving up the search as
everyone needs a mission in life *laugh*


Conrad

------------------------------------------------------------------------
-----------
"Any sufficiently advanced technology is indistinguishable from magic"
----------------------------------------------------------- Arthur C
Clarke ----





From: Kevin Mackenzie :      nhi691-at-abdn.ac.uk
Date: Tue, 8 Jul 1997 08:51:14 +0100 (BST)
Subject: Poly-L-lysine problem

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Hi

I am trying to attatch various small samples (Diatoms, Microspores and
Cecaria) onto Poly-L-lysine coated coverslips, but am finding that the
samples do not attach.

Does anyone have any ideas?

My Method

Clean coverslips in 100% ethanol
Air dry
0.1% Poly-L-lysine (MW60000) in distilled water for 1minute
Rinse in distilled water
Drop of fixative containg sample (30 - 60 minutes)
rinse, dehydrate and CPD

I also remember reading somewhere about using Poly-D-lysine instead,
has anyone tried this?

Thanks

Kevin Mackenzie
Tillydrone E.M. Unit
University of Aberdeen
Tillydrone Avenue
Aberdeen
AB9 2NT

Tel 01224-272847
Fax 01224-272396
Web site- http://www.abdn.ac.uk/~nhi691/





From: Milos Motejl :      motejl-at-paru.cas.cz
Date: Tue, 8 Jul 1997 11:26:05 +0200 (MET DST)
Subject: Poly-L-lysine problem

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unsubscribe microscopy motejl-at-paru.cas.cz





From: Ian Bache :      icb1000-at-cam.ac.uk
Date: Tue, 08 Jul 1997 10:35:42 +0100
Subject: Re: ESEM EDS ?

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Stuart McKernan wrote:

} Responding to the message of {199707072115.AA24968-at-relay.ppco.com}
} from gllovel-at-ppco.com (Gary Lovell):

} } I have been trying to identify ESEM EDS resolution using a 50 micron
} } diameter Nickel standard embedded in epoxy (C, O and Cl). My instrument
} } parameters are 1: 15kv 2: 12.0 mm working distance 3: Chamber
} } pressure=3.5T 4: Chamber Gas = N2 or H2O (did'nt make any difference which
} } gas was used) 5: Condensor setting = 50% 6: Sample tilt = 30, 20, 10.
} } Doing a spot analysis in the center of the Ni standard showed abundant C and
} } O, and some Cl as well as the expected Ni counts. Is the skirting effect of
} } the ESEM such that one can't perform EDS on smples of 50 microns or less? I
} } would be interested in hearing from anyone who has any results for ESEM EDS
} } resolution.
} }
} }
} } From the literature on this subject I think that the chamber pressure is too
} high; resulting in too much scattering of the incoming beam. There are several
} papers by Brendon Griffin in Australia, and by the Danish group in RISO -
} Horsewell, Appel and Bilde-Sorensen - where they have extensively documented the
} EDS resolution as a function of voltage, pressure and working distance. See MSA
} proceedings from 1995 and 1996 for papers from the ESEM symposia which cover
} this topic
}
} Good luck,
}
}
Our group in Cambridge have also been investigating this, and will be
presenting a paper at this years MSA conference in Cleveland :-

'Variations in the probe beam broadening with operating conditions in
ESEM'

Tuesday 11:45 - Rm. 206


Ian Bache
Research Student
Polymers & Colloids Group, Cavendish Laboratory
Cambridge University, Madingley Road, Cambridge CB3 0HE
UK




From: chris gilpin :      cgilpin-at-fs1.sem.man.ac.uk
Date: Tue, 8 Jul 1997 12:32:53 BST
Subject: Re: ESEM EDS ?

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You may also like to read my modest contribution

SIGEE, D.C. and GILPIN, C.J. (1994)
X-ray microanalysis with the environmental scanning electron
microscope: Interpretation of data obtained under different
atmospheric conditions. Scanning Microscopy Supplement 8: 219-229


1 GILPIN, C.J. and SIGEE, D.C. (1995)
X-ray microanalysis of wet biological specimens in the environmental
scanning electron microscope 1. Reduction of specimen distance under
different atmospheric conditions. Journal of Microscopy 179: 22-28


Chris

Chris Gilpin
Biological Sciences Electron Microscope Unit
G452 Stopford Building
Oxford Road
Manchester
M13 9PT
phone +44 161 275 5170
fax +44 161 275 5171
http://www.biomed.man.ac.uk/biology/emunit/emhome.html




From: jss :      jss-at-siva.bris.ac.uk
Date: Tue, 08 Jul 1997 13:16:51 +0000
Subject: EDX at High Pressure

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We do both theoretical and experimental work on high pressure SEM
microscopy . (This is a noncommercial name for ESEM.) We have been
looking at the effect of water vapour and water liquid layer on the
emission characteristic x-rays. We have recently deduce from our Monte
Carlo calculations that the true contribution from the 'image' area can
be estimated by applying a correction factor. 'Resolved image area' in
turn depends upon the thickness of the water layer or the pressure of
the water vapour above the specimen. The exact proportion of the
characteristic x-rays coming from the actual region being analysed
increases with the 'increase' in the resolavable radius. Thus the value
of the correction factor reduces as the resolution worsens. These
results and others will be presented at the Meeting MSA.

Jitu Shah
H.H. Wills Physics Laboratory,
University of Bristol
Royal Fort.
Bristol BS 8 1TL
UK
email: jss-at-siva.bristol.ac.uk


} Stuart McKernan wrote:
}
} } Responding to the message of {199707072115.AA24968-at-relay.ppco.com}
} } from gllovel-at-ppco.com (Gary Lovell):
}
} } } I have been trying to identify ESEM EDS resolution using a 50 micron
} } } diameter Nickel standard embedded in epoxy (C, O and Cl). My instrument
} } } parameters are 1: 15kv 2: 12.0 mm working distance 3: Chamber
} } } pressure=3.5T 4: Chamber Gas = N2 or H2O (did'nt make any difference which
} } } gas was used) 5: Condensor setting = 50% 6: Sample tilt = 30, 20, 10.
} } } Doing a spot analysis in the center of the Ni standard showed abundant C and
} } } O, and some Cl as well as the expected Ni counts. Is the skirting effect of
} } } the ESEM such that one can't perform EDS on smples of 50 microns or less? I
} } } would be interested in hearing from anyone who has any results for ESEM EDS
} } } resolution.
} } }
} } }
} } } From the literature on this subject I think that the chamber pressure is too
} } high; resulting in too much scattering of the incoming beam. There are several
} } papers by Brendon Griffin in Australia, and by the Danish group in RISO -
} } Horsewell, Appel and Bilde-Sorensen - where they have extensively documented the
} } EDS resolution as a function of voltage, pressure and working distance. See MSA
} } proceedings from 1995 and 1996 for papers from the ESEM symposia which cover
} } this topic
} }
} } Good luck,
} }
} } Ian Bache wrote:
} Our group in Cambridge have also been investigating this, and will be
} presenting a paper at this years MSA conference in Cleveland :-
}
} 'Variations in the probe beam broadening with operating conditions in
} ESEM'




From: jss :      jss-at-siva.bris.ac.uk
Date: Tue, 08 Jul 1997 13:16:51 +0000
Subject: EDX at High Pressure

Contents Retrieved from Microscopy Listserver Archives
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We do both theoretical and experimental work on high pressure SEM
microscopy . (This is a noncommercial name for ESEM.) We have been
looking at the effect of water vapour and water liquid layer on the
emission characteristic x-rays. We have recently deduce from our Monte
Carlo calculations that the true contribution from the 'image' area can
be estimated by applying a correction factor. 'Resolved image area' in
turn depends upon the thickness of the water layer or the pressure of
the water vapour above the specimen. The exact proportion of the
characteristic x-rays coming from the actual region being analysed
increases with the 'increase' in the resolavable radius. Thus the value
of the correction factor reduces as the resolution worsens. These
results and others will be presented at the Meeting MSA.

Jitu Shah
H.H. Wills Physics Laboratory,
University of Bristol
Royal Fort.
Bristol BS 8 1TL
UK
email: jss-at-siva.bristol.ac.uk


} Stuart McKernan wrote:
}
} } Responding to the message of {199707072115.AA24968-at-relay.ppco.com}
} } from gllovel-at-ppco.com (Gary Lovell):
}
} } } I have been trying to identify ESEM EDS resolution using a 50 micron
} } } diameter Nickel standard embedded in epoxy (C, O and Cl). My instrument
} } } parameters are 1: 15kv 2: 12.0 mm working distance 3: Chamber
} } } pressure=3.5T 4: Chamber Gas = N2 or H2O (did'nt make any difference which
} } } gas was used) 5: Condensor setting = 50% 6: Sample tilt = 30, 20, 10.
} } } Doing a spot analysis in the center of the Ni standard showed abundant C and
} } } O, and some Cl as well as the expected Ni counts. Is the skirting effect of
} } } the ESEM such that one can't perform EDS on smples of 50 microns or less? I
} } } would be interested in hearing from anyone who has any results for ESEM EDS
} } } resolution.
} } }
} } }
} } } From the literature on this subject I think that the chamber pressure is too
} } high; resulting in too much scattering of the incoming beam. There are several
} } papers by Brendon Griffin in Australia, and by the Danish group in RISO -
} } Horsewell, Appel and Bilde-Sorensen - where they have extensively documented the
} } EDS resolution as a function of voltage, pressure and working distance. See MSA
} } proceedings from 1995 and 1996 for papers from the ESEM symposia which cover
} } this topic
} }
} } Good luck,
} }
} } Ian Bache wrote:
} Our group in Cambridge have also been investigating this, and will be
} presenting a paper at this years MSA conference in Cleveland :-
}
} 'Variations in the probe beam broadening with operating conditions in
} ESEM'




From: George.C.Ruben-at-Dartmouth.EDU (George C. Ruben)
Date: 08 Jul 97 08:14:08 EDT
Subject: Re: Poly-L-lysine problem

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Why would a silica shell of a diatom stick to a positively charge surface----
You can collect diatoms on a 0.22 micron or 0.45 micron silver filter sold by
the electron microscopy houses. These filters are conductive if you are going
to look at the diatoms in SEM.

George C. Ruben
Dept. Biological Sciences
Dartmouth College
Hanover, NH USA




From: Robert A. CARLTON 610-454-3949 :      CARLTRA-at-rpr.rpna.com
Date: Tue, 08 Jul 1997 08:32:00 -0400 (EDT)
Subject: Re: ESEM-EDS

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Mr-Received: by mta FWVX01; Relayed; Tue, 08 Jul 1997 08:37:27 -0400
Alternate-Recipient: prohibited
Disclose-Recipients: prohibited

Gary,

I noticed a couple of things about your conditions which are probably
not optimum for the best resolution. I assume you are using one of
the Electroscan instruments. If so, you can reduce the beam skirt by
using the long assembly secondary electron detector (17 mm) and a
working distance of 19 mm. That leaves a 2 mm gas path length. Also,
the voltage to the secondary detector has a profound effect on many of
the measurement parameters such as the beam current (see Wight in 1996
MSA proceedings p838-839). Reducing that voltage probably has a
beneficial effect As was mentioned previously, the size of the skirt
is directly related to the gas pressure and one wants to operate as
low as possible for the samples being examined. I use 2.0 torr for my
polymer samples and lower for others. Also, higher accelerating
voltages reduce the skirt size but at an obvious cost if doing EDS of
light elements.

Notice how cleverly I gave an answer without answering your question?
I should have been a lawyer or better a politician! Frankly, I don't
think the final word has been written on EDS-ESEM resolution. The
original work seemed to indicate that we should be talking millimeters
not micrometers. More recent work, however, suggests that maybe
micrometers are appropriate with sufficient care. There is a session
on Tuesday morning on the ESEM at the MSA/MAS meeting and there will
surely be more discussion of this issue there and in the literature.

By the way, I have been trying to locate the articles by Horsewell,
Appel and Bilde-Sorenson mentioned by Stuart McKernan with no success.
Does anyone have any suggesstions on how to get ahold of them?

Robert Carlton
Rhone-Poulenc Rorer





From: leibest-at-acpub.duke.edu (Leslie Eibest)
Date: Tue, 8 Jul 1997 08:50:28 -0500
Subject: Re: Help on SEM sample preparation

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Dear Yuhui;
For particulate samples that cannot be air dried, I sandwich the
sample between two Millipore filters held in a Millipore Swinney stainless
steel filter holder. It is designed to fit on a syringe, but I cut the
ends off (which allows for good fluid exchange), and run the holder through
the dehydration series and CPD. Alternatively, some samples can be dried
in HMDS or TMS, and you can skip the critical point dryer.
Also, I recomend using conductive carbon tabs for your adhesive.
They give a nice smooth background, and are more conductive than
double-stick tape (mine came from Pella, but other companies may also
offer them).
Please feel free to contact me off-line for more details if necessary.

Leslie Eibest
Zoology Dept., Box 90325
Duke University
Durham, NC 27708 USA
(919) 684-2547
leibest-at-duke.edu






From: ROMEO Michelangelo :      romeo-at-maiorana.u-strasbg.fr
Date: Wed, 10 Feb 1993 21:22:55 +0100 (MET)
Subject: Unsubscribe

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From: William Tivol :      tivol-at-wadsworth.org
Date: Tue, 8 Jul 1997 08:55:12 -0500 (EDT)
Subject: Re: Questions on device failure analysis

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} Assuming there is a device built on a 1cm by 1cm Si wafer and the
} device has several 1-2nm thick layers and about 0.2um wide circuit
} "wiring", how can you find failure points, preferably without breaking
} the device? I know Sandi is doing X-TEM of devices and am really curious
} about the way to find, say, a short circuit on the device.
} Any comments are greatly appreciated.
}
Dear Chao-Ying,
What are the compositions of the parts of the device you want to
examine? That is, are the "wires" also mainly Si, are they a different
material with similar Z--i.e. aluminum--or do they have much different
Z? What I'm really asking is if there is a contrast-producing effect
in the device. If all the device is Si with small amounts of various
dopants for the different parts, you will have a difficult time, but if
not, either imaging or element mapping could give you the info you want.
Other questions are what resolution is necessary and what is the total
thickness of the device--is there a thick backing?
Yours,
Bill Tivol




From: Panjikar Santosh Kumar :      kumar-at-uni-muenster.de
Date: Tue, 8 Jul 1997 15:11:15 +0200
Subject: unsubscribe

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unsubscribe microscopy kumar-at-uni-muenster.de




From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Tue, 8 Jul 1997 08:29:33 -0500
Subject: Biological vs. Metallurgical EM

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I have worked in biological EM for 13 years now, but am curious about
metallurgical EM in Eng. and am wondering how difficult it would be for
me to make a switch, because I've noticed that there seems to be more
jobs for EM technologists in metallurgy than biology.

Are there any people out there that have switched from one side to the
other? I think that looking after the microscope would be essentially
the same, but I have no experience in electro-polishing and sample
preparation for metallurgy. How difficult is this?

Garry




From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Tue, 8 Jul 1997 08:33:10 -0500
Subject: MDS

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I am interested in hearing from anyone doing diagnostic EM with a
company called MDS. We are about to be privatized here, but I have no
idea what this company plans to do with electron microscopy. All the
information that I have regarding MDS basically only applies to their
high volume "core-lab" but not to their small specialized labs.

Garry




From: Stuart McKernan :      mckernan-at-cems.umn.edu
Date: Tue, 8 Jul 1997 08:37:57 -0500
Subject: Re: ESEM-EDS

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Responding to the message of {01IKZKXJZCKW90OHU4-at-mr.fwvx03.com}
from "Robert A. CARLTON 610-454-3949" {CARLTRA-at-rpr.rpna.com} :
}
} By the way, I have been trying to locate the articles by Horsewell,
} Appel and Bilde-Sorenson mentioned by Stuart McKernan with no success.
} Does anyone have any suggesstions on how to get ahold of them?
}
The references I have are all to conference proceedings:
MSA 1996 p847, Scandem 1996 Aarhus Denmark, Scandem 1997 Goteborg Sweden (and
EUREM 1996 Dublin Ireland - proceedings on CR-ROM; incomplete and virtually
useless!)

__________________
Stuart McKernan stuartm-at-tc.umn.edu
Microscopy Specialist
CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594
100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590





From: Sara Miller :      saram-at-acpub.duke.edu
Date: Tue, 8 Jul 1997 09:58:02 -0400 (EDT)
Subject: Re: Poly-L-lysine problem

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On Tue, 8 Jul 1997, Kevin Mackenzie wrote:

} Date: Tue, 8 Jul 1997 08:51:14 +0100 (BST)
} From: Kevin Mackenzie {nhi691-at-abdn.ac.uk}
} To: Microscopy-at-sparc5.microscopy.com
} Subject: Poly-L-lysine problem
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Hi
}
} I am trying to attatch various small samples (Diatoms, Microspores and
} Cecaria) onto Poly-L-lysine coated coverslips, but am finding that the
} samples do not attach.
}
} Does anyone have any ideas?
}
} My Method
}
} Clean coverslips in 100% ethanol
} Air dry
} 0.1% Poly-L-lysine (MW60000) in distilled water for 1minute
} Rinse in distilled water
} Drop of fixative containg sample (30 - 60 minutes)
} rinse, dehydrate and CPD
}
} I also remember reading somewhere about using Poly-D-lysine instead,
} has anyone tried this?
}
} Thanks
}
} Kevin Mackenzie
} Tillydrone E.M. Unit
} University of Aberdeen
} Tillydrone Avenue
} Aberdeen
} AB9 2NT
}
} Tel 01224-272847
} Fax 01224-272396
} Web site- http://www.abdn.ac.uk/~nhi691/
}
SAMPLES THAT ARE FIXED FREQUENTLY DON'T LIKE TO STICK TO ANYTHING ELSE.
TRY COATING YOUR SURFACE (WE USED 1% PL LYS FOR 10 MIN, RINSE, AIR-DRY),
ADDING YOUR SAMPLE (LET A ROUNDED DROP SIT AND DON'T LET IT OVERFLOW THE
EDGE) ON THE COVERSLIP FOR 30 MIN COVERED IN A MOIST CHAMBER. THEN
REMOVE THE EXCESS FLUID WITH A PASTEUR PIPET AND *!*GENTLY*!* ADD
FIXATIVE TO ONE SIDE AND LET IT SIT IN A ROUNDED UP DROP FOR 10-20 MIN.
WASH WITH BUFFER (GENTLY) AND PROCEED WITH WHATEVER ELSE YOU WANT TO DO.

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: Joergen Bilde-Soerensen 5802 :      j.bilde-at-risoe.dk
Date: Tue, 08 Jul 1997 16:12:46 +0200
Subject: Re: ESEM EDS ?

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Gary Lovell wrote:

} I have been trying to identify ESEM EDS resolution using a 50 micron
} diameter Nickel standard embedded in epoxy (C, O and Cl). My
} instrument parameters are 1: 15kv 2: 12.0 mm working distance 3:
} Chamber pressure=3.5T 4: Chamber Gas = N2 or H2O (did'nt make any
} difference which gas was used) 5: Condensor setting = 50% 6:
} Sample tilt = 30, 20, 10. Doing a spot analysis in the center of the
} Ni standard showed abundant C and O, and some Cl as well as the
} expected Ni counts. Is the skirting effect of the ESEM such that
} one can't perform EDS on smples of 50 microns or less? I
} would be interested in hearing from anyone who has any results for
} ESEM EDS resolution.

Dear Gary,

Approximately 75 % of the primary electrons will be scattered
when you use a working distance of 12 mm and a pressure of 3.5 torr
and a significant fraction of these scattered electrons will hit the
sample further away from the beam target than 50 micrometer. (The
scattered intensity is approximately given by Is/Io = 1 - exp(-psL/kT)
where p is the pressure, s the total scattering cross section for
electron scattering on the gas used, L the distance between the last
pressure limiting aperture and the sample, k the Boltzmann constant
and T the absolute temperature). Examples of skirt shapes are e. g.
given in:

D. A. Moncrieff et al., J. Phys. D: Appl. Phys. vol. 12 (1979)
481-88.
D. C. Joy, Microscopy and Microanalysis ' 96, Proc. Annual
Meeting MSA, Minneapolis, Minnesota, 11 - 15 August 1996

It is to a large degree possible to correct for the beam skirt
effects:

1) You can extrapolate from spectral measurements made at several
different pressures to the result that would have been found without
scattering provided that the measurements are made in the single
scattering regime (i.e. pL { approx. 1.6 Pa.m for measurements in
water vapour). In order to obtain single scattering conditions you can
use a so-called X-ray bullet to reduce the working distance.
2) If there is plural scattering, you can take two spectra, one with
a fine needle (of the kind used for field ion microscopy or scanning
tunneling microscopy) inserted over the point of interest, and the
other with the needle slightly retracted. Subtraction of the first
from the second spectrum will approximately give the spectrum from the
point of interest.

Neither method will give you as exact an analysis as you will get
under high vacuum, but you can get rid of most of the skirt effects.
The pressure variation method in particular yields pretty good results
if carefully performed. The methods are described in:

J. B. Bilde-Soerensen and C. C. Appel, Proc. 48th Annual Meeting of
the Scandinavian Society for Electron Microscopy, Aarhus, 2 - 5 June
1996, pp. 4 - 5.
J. B. Bilde-Soerensen and C. C. Appel, Proc. 11th
European Congress on Microscopy EUREM' 96, Dublin, 26-30 August 1996.
Session T6.
J. B. Bilde-Soerensen and C. C. Appel, Proc. 49th Meeting
of the Scandinavian Society for Electron Microscopy, Gothenburg, 10 -
13 June 1997, pp. 12-15.

Best wishes,
Jorgen.


J. B. Bilde-Soerensen
Materials Research Department
Risoe National Laboratory
DK-4000 Roskilde
Denmark

e-mail: j.bilde-at-risoe.dk
phone: +45 46 77 58 02 (direct)
phone: +45 46 77 46 77 (switchboard)
fax: +45 46 77 57 58
website: http://www.risoe.dk




From: Carolyn.Gondran-at-SEMATECH.Org
Date: Tue, 08 Jul 1997 09:18:07 -0500
Subject: TEM opening

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Job Title: TEM Analyst
Manager: Dr. Carolyn Gondran
Department: Materials Analysis
Division: Internal Technical Support - SEMATECH
2706 Montopolis Dr.
Austin, TX 78741-6499

(512) 356-3149 phone
(512) 356-7008 FAX

Job Summary:
Provide analytical support to SEMATECH and I300I projects and to
the ATDF through TEM analysis of semiconductor materials and devices
(in plan view and cross section) and direct interactions with
internal customers.
Operate the TEM (EDAX, STEM, PEELS, Electron diffraction etc.)
interpret electron micrographs, electron diffraction patterns and EDAX
data and provide written reports of all analyses. Work with and
oversee the activities of TEM technician(s) including selection of
sample preparation techniques, preparation of samples, photographic
image processing, maintenance of lab equipment and supplies and the
development/refinement of new sample preparation techniques as needed.

Qualifications:
An advanced degree in Physics, Chemistry or Materials Science
with proven TEM experience. 5 - 10 years of experience in TEM analysis
and sample preparation and analysis of semiconductor materials and
devices is desirable.





From: chris gilpin :      cgilpin-at-fs1.sem.man.ac.uk
Date: Tue, 8 Jul 1997 15:12:11 BST
Subject: I need a CM12

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Hi All
I have a need for a second hand Philips CM12 or CM20 preferably with
compustage. If there is anyone with one to sell or is thinking of
upgrading? I would take an instrument that is underused and give full
access to the owner if that would help.
The need is fairly urgent so a speedy reply would be appreciated even
if only to express an interest
Please reply directly to me and not the list.
Many thanks

Chris

Chris Gilpin
Biological Sciences Electron Microscope Unit
G452 Stopford Building
Oxford Road
Manchester
M13 9PT
phone +44 161 275 5170
fax +44 161 275 5171
http://www.biomed.man.ac.uk/biology/emunit/emhome.html




From: chris gilpin :      cgilpin-at-fs1.sem.man.ac.uk
Date: Tue, 8 Jul 1997 16:03:17 BST
Subject: Re: ESEM-EDS

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} As was mentioned previously, the size of the skirt
} is directly related to the gas pressure and one wants to operate as
} low as possible for the samples being examined. I use 2.0 torr for my
} polymer samples and lower for others. Also, higher accelerating
} voltages reduce the skirt size but at an obvious cost if doing EDS of
} light elements.


As people who know me will already know I am a biologist using ESEM.
I almost always view hydrated samples. Try keeping a sample wet at
2.0 Torr!
There are many people who use an ESEM to look at dry uncoated samples
and in this case there are a number of parameters available for
change.
In any general discussion on the list remember that not all samples
and imaging requirements are the same.
Sounds like lots of discussion for the ESEM session at MSA and also
the users group meeting.

Chris


Chris Gilpin
Biological Sciences Electron Microscope Unit
G452 Stopford Building
Oxford Road
Manchester
M13 9PT
phone +44 161 275 5170
fax +44 161 275 5171
http://www.biomed.man.ac.uk/biology/emunit/emhome.html




From: HILDEGARD CROWLEY :      hcrowley-at-du.edu
Date: Tue, 8 Jul 1997 09:10:24 -0600 (MDT)
Subject: Re: Good diamond knives

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Hi,
We have a large staff and many students. We have about 22 thousand
dollars worth of various kinds of diamond knives. We always buy
DIATOME. The quality is excellent and the service and support superior.
I would not consider buying any other knife from another company. In 15
years of buying and using these knives, I have never been sent a poor one
or one with any detectable flaw. I do not even "check" them out anymore
when we get one resharpened. I know it is good. (I have no stock in EMS
who sells these knives).
Sincerely,
Hildy




From: Garber, Charles A. :      cgarber-at-2spi.com
Date: Tue, 08 Jul 97 11:09:45 -0500
Subject: Mounting of powder samples

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

Kevin Mackenzie wrote:
===============================================
I am trying to attatch various small samples (Diatoms, Microspores and
Cecaria) onto Poly-L-lysine coated coverslips, but am finding that the
samples do not attach.

Does anyone have any ideas?
================================================
So long as these are relatively "free flowing" powders, larger than about 4
um, then one of our Tacky Dot (TM) Slide products should work fine in this
application. In addition, there is the added bonus that the particles are
mounted in an orthogonal array, making it possible to do analytical work on
the powder far more quickly if not also more accurately.

Information about Tacky Dot Slides can be found on our website.

Disclaimer: SPI is the sole worldwide manufacturer, under license from
DuPont, of Tacky Dot Slides so we have a vested interest in seeing that
more are used! We know of no other product like this one so there are no
other references to be given.

Chuck

===================================================
Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400
President 1-(800)-2424-SPI
SPI SUPPLIES FAX: 1-(610)-436-5755
PO BOX 656 e-mail: cgarber-at-2spi.com
West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com


Look for us!
############################
WWW: http://www.2spi.com
############################
==================================================




From: Garber, Charles A. :      cgarber-at-2spi.com
Date: Tue, 08 Jul 97 11:09:22 -0500
Subject: Cellulose acetate preparation for SEM

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

Paiboon NUANNIN wrote:
================================================
We would like to examine a pore size of cellulose acetate with SEM. Please
advise us, how to prepare the sample becuase it is very sensitive to vacuum.
Can we dry by using a CPD?
=================================================
Could you give more information about your system, for example, a) what is
the cellulose acetate "wet" with?, b) what would be the expected pore size,
and c) what is the physical form of the sample, is it a thin film coating on
a substrate or is it more of a bulk sample? With regard to cellulose
acetate, I don't think we have ever found porosity in that polymer system.
But then again, maybe we did not look hard enough either.

Chuck

===================================================
Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400
President 1-(800)-2424-SPI
SPI SUPPLIES FAX: 1-(610)-436-5755
PO BOX 656 e-mail: cgarber-at-2spi.com
West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com


Look for us!
############################
WWW: http://www.2spi.com
############################
==================================================

-------- REPLY, End of original message --------






From: Crossman, Harold :      crossman-at-OSI.SYLVANIA.com
Date: Tue, 8 Jul 1997 11:18:08 -0400
Subject: Thickness measurements

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Folks,

I have a customer who will be using SEM/EDS to characterize a mixed
oxide layer (Fe-, Ni-, Cr-, Mn-, Si-oxides, approximately 1 micrometer
thick) furnace-grown from the superalloy substrate. While measuring the
desired features will be successful at a comfortable rate in the lab,
the ultimate goal is to develop a low-prep, rapid, reliable process that
can be used on the plant floor by semi-skilled workers.

I checked around and it looks like x-ray thickness measurement tools may
do the trick. Many fluorescence units are portable, easily used, etc.
and are appropriate to our sample size (approximately 1 CM2)

My questions: Do any of you use industrial-duty x-ray fluorescence
thickness gages for measuring oxide layers over metal substrates? If
so, how accurate, repeatable, etc. Any caveats?

If you would prefer to respond directly, I can be reached at:


Harold J. Crossman
OSRAM SYLVANIA INC.
Lighting Research Center
71 Cherry Hill Dr.
Beverly, MA 01915
Phone: (508) 750-1717
E-mail: crossman-at-osi.sylvania.com

Our web sites: www.sylvania.com
www.siemens.com
--

"Crossman, Harold" {crossman-at-osi.SYLVANIA.com}





From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Tue, 8 Jul 1997 15:23:44 -0000
Subject: Amoeba is rare (Resent as missed subject line, sorry!)

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well I now know why I am having difficulty finding the Amoeba since
according to the book I have just bought, at least the classic Amoeba
Proteus is rare in the wild!!! I remember being told by a retired
microscopist that it was rare also. So at least I can take comfort in
the fact that its more than likely there are none in my samples rather
than my mis-identification! But I will not be giving up the search as
everyone needs a mission in life *laugh*


Conrad

------------------------------------------------------------------------
-----------
"Any sufficiently advanced technology is indistinguishable from magic"
----------------------------------------------------------- Arthur C
Clarke ----





From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Tue, 8 Jul 1997 14:23:47 -0000
Subject: protozoa book

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I finally got a good book on Protozoa identification. In the end I found
that the place I got my Microscope had a good book. Only 4.40 uk pounds!
Thats the kind of price I like :)

Conrad

------------------------------------------------------------------------
-----------
"Any sufficiently advanced technology is indistinguishable from magic"
----------------------------------------------------------- Arthur C
Clarke ----





From: Sally Shrom :      sally-at-retina.anatomy.upenn.edu
Date: Tue, 8 Jul 1997 12:16:17 -0400 (EDT)
Subject: Re: Good diamond knives

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Hello,
I agree completely with the praise to DIATOME. Well deserved indeed.
Sally

On Tue, 8 Jul 1997, HILDEGARD CROWLEY wrote:

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} -----------------------------------------------------------------------.
}
}
} Hi,
} We have a large staff and many students. We have about 22 thousand
} dollars worth of various kinds of diamond knives. We always buy
} DIATOME. The quality is excellent and the service and support superior.
} I would not consider buying any other knife from another company. In 15
} years of buying and using these knives, I have never been sent a poor one
} or one with any detectable flaw. I do not even "check" them out anymore
} when we get one resharpened. I know it is good. (I have no stock in EMS
} who sells these knives).
} Sincerely,
} Hildy
}





From: Ya Chen :      ychen14-at-facstaff.wisc.edu
Date: Tue, 8 Jul 1997 12:23:10 -0600
Subject: Re: Help on SEM sample preparation

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Hi YuHui,
You can try using Cryo-SEM. Put a drop of beads in suspension onto a
substrate, blot excess solution using filter paper, and fast-freeze it. In
order to expose the surface for SEM viewing, you need to etch the ice on
surface, but the bottom part of beads are still embedded in ice. Then
apply cryo-coating and cryo-observation in a cryo-SEM. If you have any
questions, please contact me offline.

Ya Chen


Ya Chen

=========================================================================
\ / Integrated Microscopy Resource (IMR)--
\ / __ an NIH Biomedical Research Resource TEL : 608-263-8481
\/ / / University of Wisconsin-Madison FAX : 608-265-4076
/ / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu
/ /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu
=========================================================================
IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html

The Integrated Microscopy Resource and Carnegie Mellon University will
be sponsoring a symposium and short course on multi-photon excitation
imaging, August 9-10, 1997, in Cleveland Ohio.






From: Greg Erdos :      gwe-at-biotech.ufl.edu
Date: Tue, 08 Jul 1997 13:55:56 -0400
Subject: Re: Good diamond knives

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We have never gotten a bad diamond knife from MicroStar in the 10 plus years
we have been dealing with them. We have at least a dozen knives and get 2
or more resharpened every year.


} } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
At 12:16 PM 7/8/97 -0400, you wrote:
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Scientific Director,
ICBR Electron Microscopy Core Lab
PO Box 118525 Fax: 352-846-0251
University of Florida E-mail: gwe-at-biotech.ufl.edu
Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
Home of the #1 Gators
*****
"Many shall run to and fro, and knowledge shall be increased"
Daniel 12:4





From: Jill Craig :      jcraig-at-unbc.edu
Date: Tue, 8 Jul 1997 13:57:01 -0700 (PDT)
Subject: fixation of diatomaceous algae

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Hi all,

We are trying to fix diatomaceous algae for SEM. Does anyone have any
suggestions or warnings they could pass along.

Specifically, we are interested in the best solution for long-term
storage (greater than 3 months).

The four suggested so far are: 1)lugol's solution, 2)formalin, 3)1%
glutaraldehyde, 3)2% paraforaldehyde EM grade, and 4)a combination of 1%
glutaraldehyde and .1% paraformaldehyde.

If you would like to post directly to me, I would be happy to submit a
summary to the group.

Thank you very much for your suggestions

Cheers,


Jill




From: Berta, Yolande :      YBerta-at-ms-mail.chemse.gatech.edu
Date: Tue, 08 Jul 97 17:30:00 EDT
Subject: 5K run/walk in Atlanta '98

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Dear Microscopists,
For the Microscopy and Microanalysis '98 meeting in Atlanta, the local
arrangements committee is seeking a volunteer to organize a fun 5K run/walk
for Sunday morning, July 12, 1998. For further information, please contact
me off-line, at my e-mail address:
yolande.berta-at-mse.gatech.edu
Yolande Berta
Georgia Tech
School of Materials Science and Engineering
778 Atlantic Dr.
Atlanta, GA 30332-0245
(404)894-2545




From: Caroline Schooley :      schooley-at-mcn.org
Date: Tue, 8 Jul 1997 15:49:37 -0700
Subject: Re: protozoa book

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What is the book? Perhaps I should put it in the Project MICRO bibliography.

Caroline


Caroline Schooley
Educational Outreach Coordinator
Microscopy Society of America
Box 117, 45301 Caspar Point Road
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html
Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/






From: W.Jablonski-at-csl.utas.edu.au (Wis Jablonski)
Date: Wed, 09 Jul 1997 09:20:10 +1100
Subject: ESEM/EDS

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My two cents worth on EDS at elevated pressures,
I published results of quantitative analysis of 70 microns Cr-spinel in
Mg-olivine matrix at pressure in the specimen chamber from 1 Tr to 16.6 Tr
with still good GSE resolution. Even at 10,000x magnification and analytical
window width 4.5 microns, there is small but significant contribution from
Mg-olivine towards Cr-spinel composition. And yes, changing working distance
(shorter) and accelerating voltage (higher) can minimise skirt. My advice is
if you have to do it in ESEM, go to the lowest possible pressure in the
specimen chamber to avoid charging and try WD and kV for the best results.
For biological specimen use a cold stage and keep a specimen at minimum
temp. close to 0 deg. In this case you will still deal with a fully hydrated
specimen at relatively low pressure of water vapor in the specimen chamber (
4.647 Tr at 0.2 deg. C)
Wis Jablonski OiC EM/X-ray Microanalysis Unit, CSL, Uni of Tasmania

Ref: W Jablonski, ESEM-2020-a key research tool in the university environment.
The Third Biennial Symposium on SEM Imaging and Analysis: Applications and
Techniques, Proceedings, Australian Microbeam Society, Feb 15-17, Melbourne
1995, pages16-17.

PS More recent and quantitative work by Bilde-Soerensen could be a good help.





From: Mary Mager :      mager-at-unixg.ubc.ca
Date: Tue, 08 Jul 1997 20:15:32 -0700
Subject: Re: Biological vs. Metallurgical EM

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Dear Garry,
If you have done biological EM then metallurgical EM should be a breeze.
Although my degree is in Microbiology, all my EM has been in Metallurgy and
Materials Engineering. There is much more SEM than TEM and a lot of EDX,
including the problems of quantitative EDX. Specimen preparation is straight
forward and can consist of just putting the metal piece on a stub and into
the SEM. You need to learn all about EDX and some basic principals of
metallurgy. Electropolishing is just recipe following and there are some
neat instruments, like ion beam thinners, to help you. It depends on what
the particular lab you work at specializes in. I don't think there is
anything as difficult as biological TEM specimen preparation and
ultramicrotoming in the metallurgical field. The trick is to get the
training you need from someone who knows it well.

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

Mary Mager
Electron Microscopist
Metals and Materials Eng., UBC
6350 Stores Rd.
Vancouver, B.C. V6T 1Z4
CANADA
tel:604-822-5648, fax:604-822-3619
e-mail: mager-at-unixg.ubc.ca





From: W.Jablonski-at-csl.utas.edu.au (Wis Jablonski)
Date: Wed, 09 Jul 1997 14:13:38 +1100
Subject: ESEM/EDS

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Further to my mail on EDS at elevated pressures:
I would like to clarify the sentence No2: Even at 10,000x magnification
and analytical window width 4.5 microns, there is a small but significant
contribution from Mg-olivine towards Cr-spinel composition at the lowest (
in this case) 1 Tr specimen chamber pressure.
Hope this will help.
WJ





From: Ritchie Sims :      r.sims-at-auckland.ac.nz
Date: Wed, 9 Jul 1997 16:28:45 GMT+1200
Subject: Re: Indium foil source

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} Help! I need a source for indium foil. I need to use this to pick up
} particles and residue from the equipment in our wafer fab.

Try ESPI, phone toll free (800) 638-2581, fax (818) 889-7098, at 5310
Derry Avenue, Agoura, CA 91301. Their catalog lists 9 different
thicknesses each in 3 purity grades!!

I have no connection with them, I merely drool over their catalog
occasionally.

Ritchie




Ritchie Sims phone: 64 9 3737599 ext 7713
Department of Geology fax: 64 9 3737435
University of Auckland
Private Bag 92019
Auckland
New Zealand




From: Stephan Coetzee :      stephan-at-gecko.biol.wits.ac.za
Date: Wed, 9 Jul 1997 07:22:58 GMT+2
Subject: Re: Help on SEM sample preparation

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I have to admit a very need solution. Being restricted on budget
AND the exchange rate! (totally unfair) We are using a cheap
solution. We produce envelopes from a ~ 5cm x 3cm piece of lint free
paper (lens paper), staple it closed and CPD the normal way.

} For particulate samples that cannot be air dried, I sandwich the
} sample between two Millipore filters held in a Millipore Swinney stainless
} steel filter holder. It is designed to fit on a syringe, but I cut the
} ends off (which allows for good fluid exchange), and run the holder through
} the dehydration series and CPD. Alternatively, some samples can be dried
} in HMDS or TMS, and you can skip the critical point dryer.
} Also, I recomend using conductive carbon tabs for your adhesive.
} They give a nice smooth background, and are more conductive than
} double-stick tape (mine came from Pella, but other companies may also
} offer them).
} Please feel free to contact me off-line for more details if necessary.
##
[########]
##
##
##
##
##
Stephan H Coeztee
Electron Microscope Unit
Private Bag 3
Wits
2050
South Africa

Stephan-at-Gecko.biol.WITS.ac.za

Tell: +27 11 716 2419
Fax : +27 11 339 3407




From: Laurent.NORMAND-at-ifp.fr
Date: 9 Jul 1997 07:27:26 +0000
Subject: Gels with TEM

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G'day,

Do any of you have hints and advises to give me about studies of gels with TEM ?
1) What kind of preparation would you use ? 2) What would you characterise first
? 3) Would try to go for cryo TEM ? 4) Would you do replica or try to use a cold
stage ?...
Your experience is more than welcommed !
Thanks.





From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Wed, 9 Jul 1997 09:18:04 -0000
Subject: protozoan book

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Message-ID: {c=US%a=_%p=Historical_Colle%l=IT3NT-970709091804Z-154-at-it3nt.pasttimes.com}

I have made a note in my palmtop to remember to bring the details of the
protozoan book that I bought into work so I can send them to this list.

I can't say how good it is when compared to other possible books due to
not having seen other books to compare it with! but I thought my little
Observers Book was good value for 2 UK pounds and that book was
concerned mainly with pond life in general. This book costs just over
twice as much and is full of detailed pictures of various protozoan
including of course several types of Amoeba which I am currently
searching for...!

It is quite interesting though how different pictures in books can be
with respect to certain creatures since I think the little observers
book has a better drawing of COLEPS than this protozoan book in my
opinion as it resembles a 'knurled barrel' whereas in this protozoan
book it doesn't give so much an impression of a 'knurled' surface which
is how it appeared to me under the 'scope!

I liked the warning about culturing Amoeba in that if you do it at too
high a temperature you can favour the culture of a LETHAL PATHONOGENIC
species of amoeba!!! although it says if careful it is unlikely as the
temperatures it quotes are above 35C which is hot for a country like the
UK!!! unless of course the central heating is turned up.....

For any amateurs on this list in the UK I obtained this book from Brunel
Microscopes for the cost of 4.40 UK Pounds.

Conrad

------------------------------------------------------------------------
-----------
"Any sufficiently advanced technology is indistinguishable from magic"
----------------------------------------------------------- Arthur C
Clarke ----





From: Pavel HOZAK :      hozak-at-sun1.biomed.cas.cz
Date: Wed, 09 Jul 1997 10:20:08 +0200
Subject: search for Philips 400 vacuum module

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Dear all,
we are in a desperate search for an old vacuum electronic module E-U12A
Philips EM400 (Philips Cat. No. 532269514353) as our old one is down and we
cannot afford to buy a new one from Philips - the old parts are
outrageously expensive. Isn't there someone owning an old Philips 400 that
is being replaced and could give us the module?

Thank you for any help or suggestion.

Pavel Hozak


__________________________

Pavel HOZAK, PhD
Inst. of Experimental Medicine
Dept. of Cell Ultrastructure & Molecular Biology
Videnska 1083
142 20 Prague 4 - Krc
Czech Republic

Tel.: (+420-2)-4752219
FAX: (+420-2)-4752782
e-mail: hozak-at-biomed.cas.cz
www page of our laboratory: http://uemweb.biomed.cas.cz/hozak.htm




From: METENGR-at-aol.com
Date: Wed, 9 Jul 1997 08:25:19 -0400 (EDT)
Subject: Re: Indium foil source

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Becky,

An excellent source for any metal is Alfa/AESAR (Johnson Matthey), they have
indium foil and many other hard to find metals.

Their info. is as follows:

Alfa Aesar
30 Bond Street
Ward Hill, MA 01835-8099

WWW -at- {http://www.alfa.com
Phone: 800-343-0660 or 508-521-6300
FAX: 800-322-4757
e-mail: catalog-at-alfa.com

I hope this helps.

Laura L. Estok
Asst. to the President
M.E. Taylor Engineering, Inc.
21604 Gentry Lane
Brookeville, MD 20833
Phone: 301-774-6246 * FAX: 301-774-6711 * e-mail: Metengr-at-aol.com







From: Marcelo Henrique Prado da Silva :      prado-at-METALMAT.UFRJ.BR
Date: Wed, 9 Jul 1997 10:01:53 EST3EDT
Subject: Biological Vs. Metallurgical EM

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Dears Garry, Mary Mager and all,

I'm a Metallugical engineer taking Ph.D. course on Biomaterials.
My thesys is on titanium implants for odontology. My M.Sc. thesis
was on a beta titanium alloy for aircraft industry and I did TEM
characterization of this alloy. Now, I'm on the opposite way: I
need to do histological cuts and analisys of the specimens (titanium
implants inserted into the tibiae of rabbits).

I'd like you to give me some information about the interpretation
of histological specimens on laser confocal microscopy or optical
microscopy. First, about specimens preparation: We have a diamond
wheel machine (ISOMET). I intend to embed the specimen with resin
and gently cut (low speed, low weight). To obtain a histological
slice, must I grind it? Till how many microns? Second: How can I
quantitatively characterize de degree of osseointegration?

I hope you can help me.

Yours sincerely,

Marcelo Henrique Prado
PEMM - COPPE/UFRJ
Po.Box.:68505
Cidade Universitaria - Ilha do Fundao
Rio de Janeiro-R.J.
CEP.: 21941-900

TEL.: 280-7443/590-2663 R.217
FAX.: 290-6626




From: Bob Lawrence :      Bob_Lawrence-at-latgqmg.sps.mot.com
Date: 9 Jul 1997 06:09:57 -0700
Subject: Indium foil

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REGARDING Indium foil

Beckey,

The best source is the Indium Corporation of America, 1-800-4-indium.
I've used the foil to capture particles for years
with excellent results. I work as a semiconductor failure analyst, 19 years
here at Motorola.

Hope this is the information you need. Have a nice day!

Respectfully,
Bob Lawrence





From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Wed, 9 Jul 1997 14:44:10 -0000
Subject: protozoa book

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Message-ID: {c=US%a=_%p=Historical_Colle%l=IT3NT-970709144410Z-873-at-it3nt.pasttimes.com}

Hi,
I hope that the I didn't imply that the little book that I have bought
had photographs in it! It has only drawings but still at 4.40 its good
value I think. I intend to post the book details when I remember to
bring them in!

Conrad

------------------------------------------------------------------------
-----------
"Any sufficiently advanced technology is indistinguishable from magic"
----------------------------------------------------------- Arthur C
Clarke ----





From: Kenneth JT Livi :      klivi-at-jhu.edu
Date: Wed, 09 Jul 1997 09:45:43 -0400 (EDT)
Subject: Re: Biological vs. Metallurgical EM

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Mary and Garry wrote about switch from Biological to Metallurgical EM:

Although I agree that sample prep can be far more demanding with biological
specimens, as far as TEM is concerned, one thing biologists will most
likely lack is training in crystallography. Most biologists I've met are
not fond of reciprocal space. Garry, if you want to switch to crystalline
solids, take a few courses in crystallography. Materials Science is far
more than just generating a conventional TEM image.

Ciao for now,
Ken

Kenneth JT Livi
Department of Earth and Planetary Sciences
34th and Charles Streets
Johns Hopkins University
Baltimore, Maryland 21218 USA
Phone: (410) 516-8342
Fax: (410) 516-7933
e-mail: klivi-at-jhu.edu






From: Ronnie Houston :      rhh1-at-airmail.net
Date: Wed, 09 Jul 1997 09:30:00 -0700
Subject: PHOSPORWOLFRAM ACID

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Can any of our German colleagues help with translation please? Came
across the term PHOSPHORWOLFRAM acid in a technical paper. Suspect it is
phosphoric acid but want to be sure before proceeding.
Thanks in advance
Ronnie Houston
Texas Scottish Rite Hospital for Children
Dallas




From: Mark E. Darus (216) 266-2895 General Electric Co. :      darus-at-cle.dnet.ge.com
Date: Wed, 9 Jul 97 10:49:23 EDT
Subject: RE: PHOSPORWOLFRAM ACID

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Look to number 7339 if you have the 11th Edition of the Merck Index.

Phosphotungstic Acid (Tungstophosphoric Acid) 24WO3.2H3PO4.48H2O

Page 7341




From: Gib Ahlstrand :      giba-at-puccini.crl.umn.edu
Date: Wed, 9 Jul 1997 10:08:31 -0500
Subject: Re: PHOSPORWOLFRAM ACID

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In message {33C3BC88.5577-at-airmail.net} writes:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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}
} Can any of our German colleagues help with translation please? Came
} across the term PHOSPHORWOLFRAM acid in a technical paper. Suspect it is
} phosphoric acid but want to be sure before proceeding.
} Thanks in advance
} Ronnie Houston
} Texas Scottish Rite Hospital for Children
} Dallas

Ronnie, wolfram in the old Germanic (I think) name for tungsten, so you have
phosportungstic acid there, which when dissolved in water to a few percent is
good ol' PTA, commonly used as a negative stain for viruses, bacteria,
particulates in transmission electron microscopy.




--

Gib Ahlstrand, MMS Newsletter Editor
Electron Optical Facility, University of Minnesota, Dept. Plant Pathology
495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249
612-625-9728 FAX, giba-at-puccini.crl.umn.edu

Plato: "When the mode of the music changes, the walls of the city will shake."

Chuck Berry: "There's a whole lotta shakin' goin' on!"





From: jeharper-at-amoco.com
Date: 7/6/97 10:23 AM
Subject: Re: EDX - Spot Mode

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I have used spot modes to do EDX "maps" on the fly.

If I suspect a high concentration of a particular element in a specific local
area of an image, I start acquiring and then move the spot onto and away from
the feature of interest while simultaneously watching the growth of a peak for
the element of interest. It is a crude but effective way of matching the
location of particular to features observed.

Jim Harper
Amoco Fabrics and Fibers

______________________________ Reply Separator _________________________________


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} I travel the world teaching practical electron microscopy and it worries me
} the emphasis that people place on "Spot Mode". I do not teach the use of
} spot mode for the following reasons-
}
} 1. Just because the spot appears in a certain position on the screen
} is this its correct position. I have found machines many microns out of
} step in X and Y directions.
}
} 2. Specimens charge and switching out of spot mode does not give the
} operator an easy opportunity to see if the spot had moved during the
} analysis.
}
} 3. Spot mode gives a false sense of analytical volume, no matter how
} small that spot may be we are almost certainly evaluating microns of
} material.
}
} Do others worry about spot mode accuracy, do others test the spot mode
} accuracy? Is it not better to simply increase the magnification watching
} the area of interest all the way up before the analysis and all the way
} down after the analysis?
}
} What do you think?

While the spot mode leaves a lot to be desired, particularly if the user
isn't aware of the problems, analysis in raster mode is not without its
difficulties. Most SEMs use some sort of of sychronisation of the line and
frame scan signals. If so, the analysis obtained in raster mode is biased
towards one edge and one corner of an area which may not even be identical
with the image area - the beam 'waits' at the beginning of each line and
each frame to synch, and the display may actually be blanked for a small
distance after the scan starts and before the scan ends (over scanning) to
hide image distortions arising from hysteresis, which is usually most
evident at the edges of the scans.

Get a nice dirty specimen and scan it with a coarse raster until the
contamination builds up. Now image the area at a lower mag, and look at the
contamination pattern - there will probably be a heavier contamination line
down one edge (the line scan 'wait') and a spot at one corner (the frame
'wait'). In addition, you may also be unlucky enough to find that the lines
making up the coarse raster are bent at each end - you don't see the
distortion in your images however because this part of the frame is blanked
from display, but it will contribute to an EDX analysis.

More modern SEMs have reduced these problems, but I believe that they are
still present in many.

I think that this demonstrates that whatever 'employers' might want, and
manufacturers try to provide, for anything beyond the most basic EM, you
need a skilled and trained operator with a full understanding of the
instrumentation, and the time to fully check and calibrate the machine (who
needs to be properly paid for undertaking a highly sophisticated and
skilled job). Otherwise you get results that are at best doubtful and at
worst wrong.

Regards,
Larry Stoter






From: Barbara Foster :      mme-at-mail.map.com
Date: Wed, 09 Jul 1997 13:25:43 -0700
Subject: Re: PHOSPORWOLFRAM ACID

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Message-ID: {33C3F3C7.74DD-at-mail.map.com}

Ronnie Houston wrote:
}
} ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.}
} Can any of our German colleagues help with translation please? Came
} across the term PHOSPHORWOLFRAM acid in a technical paper. Suspect it is
} phosphoric acid but want to be sure before proceeding.
} Thanks in advance
} Ronnie Houston
} Texas Scottish Rite Hospital for Children
} DallasDear Ronnie,

This sounds like a "common" rather than proper chemical name. If it
helps at all, Wolfram is the source for the chemical symbol "W", for
tungsten. I would try looking up tungsten salts of phosphoric acid.

Good luck.

Barbara Foster
Consortium President
Microscopy/Marketing & Education
53 Eton Street
Springfield, MA 01108 USA
(413)736-6931 fax: (413)746-9311 email: mme-at-map.com
(




From: rybicka-at-acsu.buffalo.edu (Krystyna K. Rybicka)
Date: Wed, 9 Jul 1997 13:44:03 -0400 (EDT)
Subject: Re:Glycogen

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Linda Barthel asked for a method to localize glycogen in cryosections. I
sent her some background information needed in the microscopic study of
glycogen, and I repeat them for all researchers interested in the subject.

The problem of glycogen is more complex than commonly appreciated, and the
understanding of this complexity is a sine qua non condition in the
microscopic study of glycogen.

Glycogen in the cell appears in the organelles, GLYCOSOMES, composed of
glycogen and enzymes involved in glycogen synthesis and degradation. The
structures stained by uranium and lead salts, and interpreted in EM as
"particles of glycogen" represent in fact the protein component of
glycosomes. Glycogen does not react with the ionic stains, but it can be
demonstrated histochemically, by the PAS technique (periodic acid - Schiff
reagent) in LM, and by the modification of this procedure (Thiery
technique) in EM.

The problem is that glycosomes are easily destroyed during tissue
processing. The most common destructive factor is the change in pH which
breaks the bond between glycogen and protein. The effect is that the
soluble protein component (enzymes) is washed out, and glycogen which is
not fixed, floats in the cell and aggregates into clumps. The acidic
treatment is inherent in the PAS procedure where periodic acid is used,
therefore, in the vertically processed slides, the unfixed glycogen often
accumulates as crescents at the bottom of the cells (the effect well known
in the classical histochemistry).

In EM the common destructive factor is uranyl acetate (strongly acidic)
used before tissue dehydration. In tissue processed without uranyl acetate
glycosomes appear intact, even after the priodic acid treatment in Thiery
technique, because the histochemical reaction is performed on sections
which are already embedded in the resin. This embedding prevents the
floating of the unfixed glycogen.

Freezing seems to be another destructive factor for glycosomes. Raether et
al, 1977 (Z.Parasitenkunde, 54, 149) used deep-freezing of Entamoeba
cultures, and found that only a few amoebae retained normal structure.
Their micrographs indicate that in the destroyed organisms the glycosomes
were destroyed.

Additional complication is that the described factors affect only free
glycosomes, whereas others, which are bound to different cell structures
remain resistant.

The review of glycosomes was published by
K.K.Rybicka, 1996, Tissue & Cell 28 (3) 253-267.

Best wishes in further study,
Krystyna






From: James.Passmore-at-corp.wrgrace.com
Date: Wed, 9 Jul 97 13:41:56 -0400
Subject: Re: PHOSPORWOLFRAM ACID

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} Can any of our German colleagues help with translation please? Came
} across the term PHOSPHORWOLFRAM acid in a technical paper. Suspect it is
} phosphoric acid but want to be sure before proceeding.
} Thanks in advance
} Ronnie Houston
} Texas Scottish Rite Hospital for Children
} Dallas

Ronnie,

"Wolfram" is the former chemical name for the element "Tungsten," hence the
symbol of "W" on the periodic table. I think it's safe to assume that
PHOSPHORWOLFRAM ACID is the same as PHOSPHOTUNGSTIC ACID.
Phosphotungstic acid is listed in references as a stain for EM work,
but I have no experience with it myself.

Jim Passmore
Analytical Chemist
Cryovac North America
Duncan, SC




From: Pat Hales :      hales-at-medcor.mcgill.ca
Date: Wed, 09 Jul 1997 13:45:15 -0700
Subject: TEM-collodion coated grids/dna

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Crystal,

This may have nothing to do with your problem and others on this listserver
may understand my story better than I do, but you may also want to note the
batch of your collodion as well as the cytochrome c. We do routine TEM
autoradiography which involves coating slides with a 1% collodion/amyl
acetate solution, then placing the sections on the slides, carbon coating,
processing the sections through autoradiography and placing grids on the
sections, then removing the collodion before scanning on the TEM. At one
point a couple years ago we ran into real problems removing the collodion
after the procedure - the sections were removed before the collodion!! After
much investigation and many trials a friend of mine finally spoke with the
chemist working at the company we bought our stock collodion solution from.
Apparently they had switched to a newer? better? safer? method of preparing
the stock collodion. We now specially order collodion prepared somehow with
ethanol and ethyl ether - and have never had a problem since!!!

Pat Hales
McGill University
Dept. of Anatomy & Cell Biology
hales-at-hippo.medcor.mcgill.ca





From: Leclerc Jean :      leclercj-at-magellan.umontreal.ca
Date: Wed, 9 Jul 1997 16:22:22 -0400 (EDT)
Subject: Pyroxylin coating

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Hi there!=88

Anyone knows anything about pyroxylin? I would like to know what it is
exactly, and if it might be more stable than some other films for coating
grids for the TEM. (Note that I'm looking at thick stuff - no thin
sections here!)

Thanks!

???????????????????????????????????????????????????????????????????????????=
??

"Life is the leading cause of Death" -B.C.

Jean Le Clerc

Institut de Recherche en Biologie Vegetale

leclercj-at-magellan.umontreal.ca
Voice: 514-277-7938
FAX: 514-277-7938 *call first*





From: bart-at-lasalle.edu
Date: Wed, 9 Jul 1997 17:05:29 -0400
Subject: SEM and Clay Mineral Prep.

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I will be purchasing a TOPCON SEM soon and I am interested in studying
clays.
I am most interested in the preparation techniques that investigators have
used
to get good pictures of kaolinite, smectite, illite, etc.

I have been using an X-ray diffractometer w/clays for years but wonder
about how
to mount oriented/unoriented specimens on those stubs.

I'd appreciate any help or references.

Hank Bart






From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Wed, 9 Jul 1997 16:29:58 -0500
Subject: RE: Good diamond knives

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I'd agree with the assessment of Diatome knives as being excellent. We
have 6 of them here, and they all seemed to be of excellent quality. Of
course, I also like Dupont and DDK.

My thoughts,

Garry

} ----------
} From: Greg Erdos[SMTP:gwe-at-biotech.ufl.edu]
} Sent: 8 July, 1997 12:55
} To: Microscopy-at-Sparc5.Microscopy.Com
} Subject: Re: Good diamond knives
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America




From: Steve Chapman :      PROTRAIN-at-CompuServe.COM
Date: Wed, 9 Jul 1997 18:01:12 -0400
Subject: SEM and Clay Mineral Prep.

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Mounting the specimen may be as simple as placing it onto a stub using a
conducting adhesive, waiting 20 minutes for it to dry, and then running
your new microscope at a low ( {5kV) accelerating voltage. Keep the
specimen as small as possible to limit contamination. Do not fall into t=
he
trap of believing that every specimen should be evaluated at 25kV. Vary
the kV to see how the information you visualise changes with beam
penetration, there is always one particular kV that will best present the=

specimen features. Obtain a simple Monte Carlo program so that you can
calculate the depth from which the backscatter and x-ray information is
coming. Changing Z and tilt will vary the backscatter contribution to yo=
ur
image and change the way the information is presented.

However if you wish to carry out EDX analysis in the microscope you will=

need up to 25kV to enable a full range spectrum evaluation. In this case=

if the material is non conducting you will need to coat it, preferably wi=
th
carbon. The simplest carbon coating systems are desk top and may either =
be
self contained or as an accessory for a sputter coater. In either case a=

carbon string system should be all that is required. The coating system i=
s
not the most important feature but the type of string you use will decide=

how easy it is to coat the specimen. We find the thick string like a boo=
t
lace is the best. If you new SEM is of the variable pressure variety th=
is
will help you reduce any problems of specimen charge

For both imaging atomic number contrast and to better visualise your
specimen for EDX evaluation you need to use a backscattered electron
detector. Varying the kV, viewing by backscatter differing depths of
information, you will effectively be able to section the specimen, from 5=
kV
through to the microscopes maximum.. The Monte Carlo program may be used=

to add penetration figures to your investigation.

Steve Chapman
Senior Consultant
Protrain




From: Roderick Ford :      Roderick.Ford-at-asu.edu
Date: Wed, 09 Jul 1997 16:05:13 -0700
Subject: Re: Electron diffraction patterns

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Diffraction pattern indexing can be difficult even if you know the
general structure, i.e. orthorhombic, because of the different length
sides of the unit cell (let alone angles in other structures). Then,
just because you can find a set of reciprocal space planes at the right
distances and angles from each other doesn't mean that the structure
factor allows for that spot to be strong in reciprocal space. So any
indexing software usually requires that you know positions of atoms in
the unit cells of all the structures that would potentially fit the
pattern that you are trying to index. EMS can be run on unix or vax
machines. You must not only know the possible structures but also a
range of camera length.

But,...I have found an alternate although cumbersome and limited
method:
I have generated and used successfully in Microsoft Excel a 3 sheet
spreadsheet that notifies you of all planes that match the input
specifications of the pattern to be indexed. You must also input the
cell parameters and cell angles. This does not take into account any
structure factors. And, it is limited by computer RAM. If you are
interested, I can tell you more how to generate it yourself -- better
that my old one.

So -- sorry, no easy indexing.




From: Sara Miller :      saram-at-acpub.duke.edu
Date: Wed, 9 Jul 1997 19:25:39 -0400 (EDT)
Subject: Re: TEM-collodion coated grids/dna

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Try a light coat of carbon .

Try Formvar.

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: Brad Goodwin :      goodwib-at-wdni.com
Date: Wed, 09 Jul 1997 16:41:34 -0700
Subject: early vs. late wood staining

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When staining sections of Douglas-fir twigs, I use safranine/fast green.
I would like to see a stain difference between early and late wood, but
with this stain I can only tell the difference by the size of the
vessels and the thickness of their walls. I would like a cleaner
method. Any ideas?




From: gcruz-at-imm.hokudai.ac.jp (Ginny Cruz)
Date: Thu, 10 Jul 1997 08:43:58 +0900 (JST)
Subject: Re: ihc on em

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Hello everyone! I'm sending this message to you in behalf of Ms. Louise Tayl
or because I believe that her question might get more replies from this netw
ork. Please send your replies to Ms Taylor directly at this address:

179LOU-at-chiron.wits.ac.za
Thanks!


At 1:51 PM 97.7.9, Ms Louise Taylor wrote:
} Hi there histonetters
}
} I have a query from an electron microscopist in our department. They
} are having some problems doing IHC on their samples, particularly
} with HMB45 (DAKO).
} Are there any suggestions out there regarding optimum dilution of the
} antibody or conversely another source that works well at EM level. I
} realise that this is not necessarily a histonet query, but I would
} appreciate whatever contacts, info etc I can get.
}
} Many Thanks
} Louise Taylor






From: Garber, Charles A. :      cgarber-at-2spi.com
Date: Wed, 09 Jul 97 22:45:07 -0500
Subject: Gels with TEM

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

Laurent NORMAND wrote:
================================================
Do any of you have hints and advises to give me about studies of gels with
TEM?
1) What kind of preparation would you use ? 2) What would you characterise
first? 3) Would try to go for cryo TEM ? 4) Would you do replica or try to
use a cold stage ?...
=================================================
There might be as many philosophies of approach as there are people doing
this kind of work. My first thought is always to determine what kind of
features are you really trying to resolve in the gel structure. For example
, certain greases (they act as gels because they have "thickeners") contain
various metal stearates that form fibrillar type networks, and the
differences between samples show up by just taking the gel, often times not
even diluting it but just smearing it out on a glass slide to the thinnest
possible film, and after RT drying, carrying out Pt/C shadowing and
replication and looking at it that way. You see the metal stearate
structures quite nicely and they usually, or at least some of them, get
picked up with the replica so you can even do ED and DF studies. And the
experimental procedure is very definitely not a complicated one, using just
conventional equipment that just about everyone has at their fingertips in
their laboratories.

On the other hand, if you have something like an aviation jet fuel kind of
gel, and one is interested in seeing the network features of the polymeric
additives (e.g. the ones that are responsible for the gelling of the system)
, freeze fracture TEM is more appropriately indicated. If you are seeking
to resolve "micropores", you might see them this way as well. If the gel,
such as a so-called "hydro gel" type system is being looked at, being
aqeuous based, some freeze etching should also be done.

If there is some kind of a non-organic type additive, that would have enough
electron density in its own right, and the goal is to see its degree of
dispersion in the gel, then cryo-TEM would be our preference. If the gel is
aqueous based, and the presence of pores is what is to be resolved, then we
have looked for ways to precipitate silver chloride into the pores to serve
as a "decorator".

The conspicuous absence of mention of SEM was not accidental. We have
almost never found SEM to be good enough to resolve the kinds of features
people want to see when doing this kind of work.

It did not sound like you are asking about things like "gel" spots in
polymer films which of course would require entirely different approaches,
none requiring cryo, except maybe for diamond knife thin sectioning.

Disclaimer: We have no connection with any of these techniques except that
we do offer them as a service for clients wanting to do this type of work.

Chuck

===================================================
Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400
President 1-(800)-2424-SPI
SPI SUPPLIES FAX: 1-(610)-436-5755
PO BOX 656 e-mail: cgarber-at-2spi.com
West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com


Look for us!
############################
WWW: http://www.2spi.com
############################
==================================================




From: Mary Mager :      mager-at-unixg.ubc.ca
Date: Wed, 09 Jul 1997 21:16:11 -0700
Subject: Re: SEM and Clay Mineral Prep.

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Dear Hank,
When I prepare clays to see the crystal structure, I usually suspend the
clay in alcohol to the consistancy of milk and then put a drop on a SEM stub
with a small glass cover slip on top, to get a very smooth surface. Dry,
then gold coat. This will allow you to see the "books" that are
characteristic of kaolinite. I use this method to look at the different
crystal shapes of kaolinite, halloysite and illite for a lab.
You wrote:
}
} I will be purchasing a TOPCON SEM soon and I am interested in studying
} clays.
} I am most interested in the preparation techniques that investigators have
} used
} to get good pictures of kaolinite, smectite, illite, etc.
}
} I have been using an X-ray diffractometer w/clays for years but wonder
} about how
} to mount oriented/unoriented specimens on those stubs.
}
} I'd appreciate any help or references.
}
} Hank Bart
Regards,
Mary

Mary Mager
Electron Microscopist
Metals and Materials Eng., UBC
6350 Stores Rd.
Vancouver, B.C. V6T 1Z4
CANADA
tel:604-822-5648, fax:604-822-3619
e-mail: mager-at-unixg.ubc.ca





From: Dr. T. J. Filler :      filler-at-uni-muenster.de
Date: Thu, 10 Jul 1997 07:55:34 +0100
Subject: New listserver

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Dear microscopists,

as it might be of interest for some of you, I wish to point
you to a new listserver at the medical faculty of the
Westfalian Wilhelms-University. It discusses anatomical
and related topics. However, it is announced for the english
and for the german language. On the other hand it has just
started and traffic is still low; therefore it might
become common to discuss in English as Germans appear sometimes
to be a bit offish :-)

To subscribe send an e-Mail to
Majordomo-at-medweb.uni-muenster.de
with the following body text:
subscribe ANATOMIE-D {your-eMail-address}

--
Mit freundlichen Gruessen Yours sincerely
********************************************************************
* Dr. T. J. Filler * specialist in anatomy *
* Westfaelische Wilhelms-Univ. * phone:*49 251 83 55226 *
* Institute of Anatomy * FAX.: *49 251 83 55241 *
* Image Analysis Division * e-Mail: filler-at- *
* Vesaliusweg 2-4 * e-Mail: image.analysis-at- *
* D-48149 Muenster Germany * e-Mail: Institute.of.Anatomy-at- *
* ______ _ ______ * domain: uni-muenster.de *
/_____/\ / /\/_____/\http://medweb.uni-muenster.de/institute/anat *
\/ /\_\// / /\/ /\_\/**********************************************
/ / / / / / / _/\
/_/ / / / / /_/\_\/
\_\/__/ / / \_\/
/___/ /
\___\/




From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Thu, 10 Jul 1997 09:07:31 -0000
Subject: apology

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Sorry if I have annoyed people but with all due respect Ray, I was asked
to share the information about this book by fellow amateurs. I admit
that I may have rambled on a bit. Although I have been on the net for a
long time, I guess with respect to this list I am one of those 'dreaded
newbies'. and thought it was like a newsgroup and forgot it was composed
mainly of proffesionals.

I only posted because it would have taken too much time to reply to
everyone and I had better end this here before this becomes another
rambling post!

Regards,

Conrad

} -----Original Message-----
} From: Ray Hicks [SMTP:rh208-at-cus.cam.ac.uk]
} Sent: Wednesday, July 09, 1997 6:54 PM
} To: Conrad Perfett
} Subject: Re: protozoan book
}
} Conrad,
}
} It's good that you've found some information useful to you, but why not
} wait until someone asks before you share it? You'll notice that there
} isn't very much spontaneous posting to the list by anyone else, about
} microscopy but especially about their research interests. Remember that
} the common interest of the list is microscopy, not microbiology, histology
} or metallurgy. If, for instance, someone has a microscopy related
} metallurgical question they post it and get an answer, generally from
} another metallurgist, you don't tend to get "I looked at a piece of
} pearlite yesterday - interesting eh?" type comments. If you did you might
} get twenty anecdotes a day from each of the members of the list, and it
} would stop being so useful.
}
} How about lurking around until someone asks a question that's in your area
} of expertise, and then helping them out?
}
} By the way, my school teachers used to provide amoebae for practical
}




From: Pavel HOZAK :      hozak-at-sun1.biomed.cas.cz
Date: Thu, 10 Jul 1997 11:15:36 +0200
Subject: search for Philips 400 vacuum module

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Dear all,
we are in a desperate search for an old vacuum electronic module E-U12A
Philips EM400 (Philips Cat. No. 532269514353) as our old one is down and we
cannot afford to buy a new one from Philips - the old parts are
outrageously expensive. Isn't there someone owning an old Philips 400 that
is being replaced and could kindly give us the module or to suggest a
solution?

Thank you for any help or suggestion.

Pavel Hozak


__________________________

Pavel HOZAK, PhD
Inst. of Experimental Medicine
Dept. of Cell Ultrastructure & Molecular Biology
Videnska 1083
142 20 Prague 4 - Krc
Czech Republic

Tel.: (+420-2)-4752219
FAX: (+420-2)-4752782
e-mail: hozak-at-biomed.cas.cz
www page of our laboratory: http://uemweb.biomed.cas.cz/hozak.htm




From: curo-at-uia.ua.ac.be (Chul-Un Ro)
Date: Thu, 10 Jul 1997 13:06:25 +0200 (MET DST)
Subject: Need help on obtaining CITZAF program

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Hello,

I am working on single aerosol particle analysis using windowless SEM/EDX
system. I hope to quantitatively analyze the contents of C, N, and O in
microparticles. CITZAF program has been known to work for particles, so I am
looking for where I could get it. I know it is a shareware and was
distributed by Caltech. However, Dr. Armstrong seems not working over there
any more. Would someone kindly help me to have a copy of CITZAF program?
Many thanks in advance.

Sincerely yours,
Chul-Un Ro

Chul-Un Ro, Ph.D
Department of Chemistry
University of Antwerp
B-2610 Wilrijk
Belgium

e-mail : curo-at-uia.ua.ac.be





From: Brad Storey :      bstorey-at-awmailhost.anlw.anl.gov
Date: 10 Jul 97 07:46:40 -0700
Subject: TEM Vibration Isolation

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Howdy,
We have a JEOL 2010 STEM that must be isolated from a huge vibrating air
compressor in a different part of the building (no they won't move it or
isolate the compressor). Our plan is to cut the floor (8" of high
density reinforced concrete sitting on dirt) around the microscope. I
am interested in tips and lessons learned,e.g., width of cut, filler
material in the cut, is it ok to tile over the gap, how far from the
scope to place the cut, etc.

Brad Storey
Materials Scientist
Argonne National Lab - West
P.O. Box 2528
Idaho Falls, ID 83403
Ph. 208-533-7685
Fax 208-533-7683
e-mail brad.storey-at-anl.gov





From: Tobias Baskin :      baskin-at-biosci.mbp.missouri.edu
Date: Thu, 10 Jul 1997 08:49:46 -0600
Subject: History...Wolfram and Tungsten

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Message-Id: {v03020900afeaa5dd8895-at-[128.206.15.200]}
Mime-Version: 1.0
Content-Type: text/plain; charset="us-ascii"

Greetings,
Any of you erudite types out there know the origins of the word
"Wolfram"? or why or when the word "tungsten" (which I believe comes from
sweedish words for "heavy + steel") came to be the prefered word? Please
forgive my curiousity if this is too far off the microscopical axis for
your taste.

Thanks,
Tobias Baskin

_ ____ ^ __ ____ Tobias I. Baskin
/ \ / / \ / \ \ University ofMissouri
/ | / / \ \ \ BiologicalSciences
/___/ /__ /___ \ \ \__ 109 Tucker Hall
/ / / \ \ \ Columbia, MO 65211 USA
/ / / \ \ \ voice: 573-882-0173
/ /____ / \ \__/ \____ fax: 573-882-0123






From: corwinl-at-pt.cyanamid.com
Date: Thu, 10 Jul 1997 09:37 -0500 (EST)
Subject: Re: Pyroxylin coating

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Short article on pyroxylin in Merck Index. Cellulose nitrate in old
film base and plastics is not stable over decades, but this probably
does not matter for your needs. Note hazards.




From: Diane Montpetit :      montpetitd-at-em.agr.ca
Date: Thu, 10 Jul 1997 09:55:54 -0400
Subject: formvar background and pta/u.a. stains

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hello everyone,

I am currently staining milk proteins with phosphotungtic acid (2%/water
ph 7.3) and uranyl acetate (1%/water) and often I do get what I called a
background consisting of very small (about 20-40nm) bubbles-like
structures that interfere with my samples...

I also recall seeing the same thing with viruses but this time it is rather
annoying because it also looks a lot like my protein...

Does somebody out there has a explanation to this phenomenon....I though
maybe it was perhaps a hydrophobic reaction between the formvar and
the stain or irregularities/defects in the formvar film....

thank you,

Diane Montpetit
Food research center
agriculture canada
st-hyacinthe, quebec, Canada
fax 514 773 8461
tel; 514 773 1105
e mail; montpetitd-at-em.agr.ca




From: Mark E. Darus (216) 266-2895 General Electric Co. :      darus-at-cle.dnet.ge.com
Date: Thu, 10 Jul 97 10:34:44 EDT
Subject: Re: History...Wolfram and Tungsten

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Tungsten

The tungsten mineral wolframite was known in the tin mines of
the Saxony-Bohemia region long before the element itself was discovered. In
1781, the Swedish chemist Scheele, who had been working with the stony mineral,
elucidated the composition of this mineral to be a compound of calcium with
an unknown acid. The acid-forming element thus discovered was named tungsten
by A. F. Cronstedt in 1755. He derived this name, from the Swedish words
"tung", meaning heavy, and "sten", meaning stone.



Wolfram

In 1783, the brothers J.J. and F. de Elhujar found that wolframite
also contained tungsten. After success in obtaining metallic tungsten from
wolframite, and gave it the name "wolfram" The origin of the word is not
clear, but it is assumed to be derived from German words "Wolf" and "Rahm" or
Swedish word "wolfrig".


Wolfram is the official international alternate name for tungsten.
Tungsten is preferred in the United States.




From: MICHAEL DELANNOY :      delannoy-at-welchlink.welch.jhu.edu
Date: Thu, 10 Jul 1997 10:49:45 -0400 (EDT)
Subject: resinless embedding

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Hello fellow microscopists,
I'm in the process of using the DGD resinless embedding
technique developed by Capco, Krochmalnic and Penman and I'm
looking for: 1)Short cuts-the ethanol, nbutanol to DGD transition
is very long (6 hours) and I am working with monolayers,
so is there a poblem with this version:
Graded ethanol dehydration to 100% 30 min
100% ethanol 4 changes 1 hr
1:1 nbutanol:100% ethanol 30 min
100% nbutanol 1 hr
1:1 nbutanol:DGD 45 min
100% DGD (with DMSO) 1 hr
RT harden O/N


2) Also any suggesstions on how to harden this wax on
tissue culture dishes, coverslips (glass) and platic slides will
be greatly appreciated. Thank you.

Improvising in Baltimore,
Mike D.




From: Pavel HOZAK :      hozak-at-sun1.biomed.cas.cz
Date: Thu, 10 Jul 1997 16:54:58 +0200
Subject: search for Philips 400 vacuum module

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Dear all,
we are in a desperate search for an old vacuum electronic module E-U12A
Philips EM400 (Philips Cat. No. 532269514353) as our old one is down and we
cannot afford to buy a new one from Philips - the old parts are
outrageously expensive. Isn't there someone owning an old Philips 400 that
is being replaced and could give us the module?

Thank you for any help or suggestion.

Pavel Hozak


__________________________

Pavel HOZAK, PhD
Inst. of Experimental Medicine
Dept. of Cell Ultrastructure & Molecular Biology
Videnska 1083
142 20 Prague 4 - Krc
Czech Republic

Tel.: (+420-2)-4752219
FAX: (+420-2)-4752782
e-mail: hozak-at-biomed.cas.cz
www page of our laboratory: http://uemweb.biomed.cas.cz/hozak.htm




From: C. John Runions :      cjr14-at-cornell.edu
Date: Thu, 10 Jul 1997 11:30:33 -0400
Subject: Re: early vs. late wood staining

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Hi Brad, I have done a bit of work staining core samples and twig sections
of Doug fir in an attempt to age the wood, and safranin staining was the
best way to evaluate this. Unfortunately, the only difference between
early and late wood is vessel diameter (ie. secondary cell wall thickness).
There may be differences in minor cell wall components from early to late
wood but none of them (I predict) would show as good a demarcation as
safranin does in staining lignin. Don't overstain with fast green as you
can lose contrast, in fact, it can be left out altogether because you are
not interested in staining cytoplasmic components. Remember that in Doug
fir there will be a gradual transition from early to late wood. You have
to estimate where the change is based on an arbitrary cell wall thickness
that you get to choose (I get a real rush making executive decisions like
that). Cheers, John

=================
C. John Runions, Ph.D
Section of Ecology and Systematics
Corson Hall
Cornell University
Ithaca, New York
USA 14853

email cjr14-at-cornell.edu [ie. cjr(fourteen)-at-...]
phone (607) 254-4282
Fax (607) 255-8088






From: C. John Runions :      cjr14-at-cornell.edu
Date: Thu, 10 Jul 1997 11:30:33 -0400
Subject: Re: early vs. late wood staining

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

Hi Brad, I have done a bit of work staining core samples and twig sections
of Doug fir in an attempt to age the wood, and safranin staining was the
best way to evaluate this. Unfortunately, the only difference between
early and late wood is vessel diameter (ie. secondary cell wall thickness).
There may be differences in minor cell wall components from early to late
wood but none of them (I predict) would show as good a demarcation as
safranin does in staining lignin. Don't overstain with fast green as you
can lose contrast, in fact, it can be left out altogether because you are
not interested in staining cytoplasmic components. Remember that in Doug
fir there will be a gradual transition from early to late wood. You have
to estimate where the change is based on an arbitrary cell wall thickness
that you get to choose (I get a real rush making executive decisions like
that). Cheers, John

=================
C. John Runions, Ph.D
Section of Ecology and Systematics
Corson Hall
Cornell University
Ithaca, New York
USA 14853

email cjr14-at-cornell.edu [ie. cjr(fourteen)-at-...]
phone (607) 254-4282
Fax (607) 255-8088






From: C. John Runions :      cjr14-at-cornell.edu
Date: Thu, 10 Jul 1997 11:39:55 -0400
Subject: Re: early vs. late wood staining

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

Hi Brad, I have done a bit of work staining core samples and twig sections
of Doug fir in an attempt to age the wood, and safranin staining was the
best way to evaluate this. Unfortunately, the only difference between
early and late wood is vessel diameter (ie. secondary cell wall thickness).
There may be differences in minor cell wall components from early to late
wood but none of them (I predict) would show as good a demarcation as
safranin does in staining lignin. Don't overstain with fast green as you
can lose contrast, in fact, it can be left out altogether because you are
not interested in staining cytoplasmic components. Remember that in Doug
fir there will be a gradual transition from early to late wood. You have
to estimate where the change is based on an arbitrary cell wall thickness
that you get to choose (I get a real rush making executive decisions like
that). Cheers, John


=================
C. John Runions
Section of Ecology and Systematics
Corson Hall
Cornell University
Ithaca, New York
USA 14853

email cjr14-at-cornell.edu [ie. cjr(fourteen)-at-...]
phone (607) 254-4282
Fax (607) 255-8088






From: Gib Ahlstrand :      giba-at-puccini.crl.umn.edu
Date: Thu, 10 Jul 1997 11:00:40 -0500
Subject: Re: formvar background and pta/u.a. stains

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In message {s3c4b19e.001-at-EM.AGR.CA} Diane Montpetit writes:
} hello everyone,
}
} I am currently staining milk proteins with phosphotungtic acid (2%/water
} ph 7.3) and uranyl acetate (1%/water) and often I do get what I called a
} background consisting of very small (about 20-40nm) bubbles-like
} structures that interfere with my samples...
}
} I also recall seeing the same thing with viruses but this time it is rather
} annoying because it also looks a lot like my protein...
}
} Does somebody out there has a explanation to this phenomenon....I though
} maybe it was perhaps a hydrophobic reaction between the formvar and
} the stain or irregularities/defects in the formvar film....
}
} thank you,
}
} Diane Montpetit
} Food research center
} agriculture canada
} st-hyacinthe, quebec, Canada
} fax 514 773 8461
} tel; 514 773 1105
} e mail; montpetitd-at-em.agr.ca

Perhaps you could modify your formvar/grid preparation techniques to see if it
makes any improvement. You could try: 1. Coat the Formvar coated grids with a
thin layer of evaporated carbon. 2. Treat the Formvar coated grids in a glow
discharge device if one is available. Either treatment may improve spreading and
negative staining of your samples.

I looked at milk proteins, too, a few years ago using PTA staining and I don't
recall any problems. The "bubbles" you see in the background may be due to
contaminant that got on the Formvar film during preparation or handling, so look
at your Formvar coating technique to see if you can clean it up; use pure double
distilled water to float film off a previously cleaned glass slide, water in
clean trough, etc, etc.

Good Luck!


--

Gib Ahlstrand, MMS Newsletter Editor
Electron Optical Facility, University of Minnesota, Dept. Plant Pathology
495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249
612-625-9728 FAX, giba-at-puccini.crl.umn.edu

Plato: "When the mode of the music changes, the walls of the city will shake."

Chuck Berry: "There's a whole lotta shakin' goin' on!"





From: hadams-at-nmsu.edu ()
Date: Thu, 10 Jul 1997 10:10:55 +0000
Subject: Re: Pyroxylin coating

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} Date: Wed, 9 Jul 1997 16:22:22 -0400 (EDT)
} From: Leclerc Jean {leclercj-at-magellan.umontreal.ca}
} To: Microscopy Association of America {Microscopy-at-Sparc5.Microscopy.Com}
} Subject: Pyroxylin coating

} ------------------------------------------------------------------------
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} -----------------------------------------------------------------------.
}
} Hi there!^
}
} Anyone knows anything about pyroxylin? I would like to know what it is
} exactly, and if it might be more stable than some other films for coating
} grids for the TEM. (Note that I'm looking at thick stuff - no thin
} sections here!)
}
} Thanks!
}
Our lab uses Pyroxylin, also known as parlodian or collodion,
frequently, as it is easy to use. We buy as a 2% solution in amyl
acetate from an EM supplier. The method: one drop on DDW in a 10mm
diameter dish, let the film dry and then remove with the tip of a
glass pipet to remove any dust particles, followed by another drop
and let dry (interference colors become visible when viewed at a low
angle). Then very gently place clean 10 to 25 grids dull side down
on the film's surface.Carefully lay a piece of parafilm on the
surface, pull the parafilm back up, and the grids plus film come
along. Place in petri dish to thoroughly dry, followed by carbon
evaporation. It is a very quick method and suitable for a number of
applications.

Hank Adams
EML
New Mexico State University






From: Diane Montpetit
Date: 10 July 1997 17:04
Subject: formvar background and pta/u.a. stains

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Diane

you didn't mention what sort of coating you are using on your grid. Could
these bubble structures be defects in the plastic coating caused by moisture
absorbed into the liquid plastic stock solution? I must admit they sound
very small but you can get small semi-perforate holes when you try to make
holey plastic films.

If they are present before sample and stain they would be more difficult to
see but should show up with a small objective aperture and lower KV.

Sorry if you've already thought of this.

Malcolm Haswell
e.m. unit
University of Sunderland
UK
----------

hello everyone,

I am currently staining milk proteins with phosphotungtic acid (2%/water
ph 7.3) and uranyl acetate (1%/water) and often I do get what I called a
background consisting of very small (about 20-40nm) bubbles-like structures
that interfere with my samples...

I also recall seeing the same thing with viruses but this time it is rather
annoying because it also looks a lot like my protein...

Does somebody out there has a explanation to this phenomenon....I though
maybe it was perhaps a hydrophobic reaction between the formvar and the
stain or irregularities/defects in the formvar film....

thank you,

Diane Montpetit
Food research center
agriculture canada
st-hyacinthe, quebec, Canada
fax 514 773 8461
tel; 514 773 1105
e mail; montpetitd-at-em.agr.ca





From: wise-at-vaxa.cis.uwosh.edu
Date: Thu, 10 Jul 1997 13:21:37 +0000
Subject: Image Pro Plus

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To all,

We have an older, DOS-based version of Image Pro Plus image
analysis software that has us stymied. We are capturing video images of
chloroplast movements and dumping them into a DOS-based computer. We want
to be able to sum all of the white areas in a field and then express that
total white area as a percentage of the total image area. It seems simple
enough, but it's not. We can identify all of the "white" objects, we can
define what we are calling 'white', we can get the area of each individual
white object, but we cannot sum the areas of all of the white objects.
Image Pro Plus corporation is no help. They sold the rights to the DOS
version to another company and no one a IPP knows anything about it. They
are quite willing to sell us the Windows version for $3500 but cannot help
us with their older version.
Does anyone out there in microscope land have experience with this
program?

Yours in computer frustration,

Bob






From: Bjorn_Bergsten-at-pei.philips.com (Bjorn Bergsten)
Date: 7/10/97 8:49 AM
Subject: History...Wolfram and Tungsten

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Dear Microscopists,

As a Swede, at least I can help with the Tungsten history:

"tung" = heavy
"sten" = stone.

Regards,

Bjorn Bergsten (= Bear Mountainstone)
EDAX International


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Greetings,
Any of you erudite types out there know the origins of the word
"Wolfram"? or why or when the word "tungsten" (which I believe comes from
sweedish words for "heavy + steel") came to be the prefered word? Please
forgive my curiousity if this is too far off the microscopical axis for
your taste.

Thanks,
Tobias Baskin

_ ____ ^ __ ____ Tobias I. Baskin
/ \ / / \ / \ \ University ofMissouri
/ | / / \ \ \ BiologicalSciences
/___/ /__ /___ \ \ \__ 109 Tucker Hall
/ / / \ \ \ Columbia, MO 65211 USA
/ / / \ \ \ voice: 573-882-0173
/ /____ / \ \__/ \____ fax: 573-882-0123






From: m.munro :      ab157-at-ab.sac.ac.uk
Date: Thu, 10 Jul 1997 20:50:57 +0100 (BST)
Subject: Confocal list

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Dear all,
Could someone please give me the address of the listserver for
confocal microscopy.

Thanks a lot,

Mark Munro
SAC Aberdeen
m.munro-at-ab.sac.ac.uk




From: Balaji Chandrasekaran :      bch605-at-hecky.acns.nwu.edu
Date: Thu, 10 Jul 1997 14:53:29 -0500 (CDT)
Subject: unsubscribe

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unsubscribe from the list server.

Thanks,
Balaji





From: Buddy Steffens :      steffens-at-calc.vet.uga.edu
Date: Thu, 10 Jul 1997 16:02:05 EST
Subject: Re: early vs. late wood staining

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Back in my early graduate school days when I used to be a botanist,
we used a stain called phloroglucinol which gave excellent
demarcation of wood based on lignin content. I remember that you
have to be very careful not to overstain, as everything will become
crimson. The rest of the tissue, parenchyma etc. was contrasted with
fast green. The stain can be modified to distinguish between
deciduous and conifer wood (excluding Ginko), although I never tried
this. The technique is described in Johansen's "Plant
Microtechnique". I'm not sure if this stain would apply to your
case, but it might be worth a try.




-=W.L. Steffens=-
Department of Veterinary Pathology
College of Veterinary Medicine
University of Georgia
Athens, GA 30602 USA
http://www.vet.uga.edu/vpp/wls/steffens.html




From: Buddy Steffens :      steffens-at-calc.vet.uga.edu
Date: Thu, 10 Jul 1997 16:08:39 EST
Subject: Re: formvar background and pta/u.a. stains

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The "bubbles" on the substrate that you describe are wetting
artifacts...at least that is what I was told by an oldtimer. I get
them consistently on negative stained viral samples, unless I
incorporated a small amount (a very small amount) of bacitracin into
the negative stain. I suppose some other surfactants would work, but
bacitracin does the trick for me every time.




-=W.L. Steffens=-
Department of Veterinary Pathology
College of Veterinary Medicine
University of Georgia
Athens, GA 30602 USA
http://www.vet.uga.edu/vpp/wls/steffens.html




From: Dr. Kristof Kovacs :      kris-at-elod.vein.hu
Date: Thu, 10 Jul 1997 22:21:06 +0200
Subject: Re: SEM and Clay Mineral Prep.

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My experience is better with isopropyl alcohol, more dilute suspension plus
in some stubborn cases a litle ultrasonic bathing... Everything will be visible!
Kris

} When I prepare clays to see the crystal structure, I usually suspend the
} clay in alcohol to the consistancy of milk and then put a drop on a SEM stub
} with a small glass cover slip on top, to get a very smooth surface. Dry,
} then gold coat. This will allow you to see the "books" that are
} characteristic of kaolinite. I use this method to look at the different
} crystal shapes of kaolinite, halloysite and illite for a lab.





From: howard-at-cshl.org (Tamara Howard)
Date: Thu, 10 Jul 1997 09:44:55 -0400 (EDT)
Subject: fluorescence?

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Disclaimer first: I'm not a chemistry/physics person, so please
excuse me if this question is incredibly stupid :)
Is it possible for a pigment to scatter/reflect light strongly
enough to look like fluorescence? If so, how could you distinguish such
behaviour from "real" fluorescence? I'm trying to work with some pigmented
plant samples that "glow" under my rhodamine/Tx Red excitation/emission
filters (standard fluorescence scope); the pigmented areas look normal
under FITC and UV conditions. The pigment itself is red under regular
light. We'd like to know if the pigment autofluoresces or if this is just
some light/pigment interaction. I'm lost and our library isn't heavy on
this type of references. Any of you "hard science" types willing to try to
educate me?!

TIA!

Tamara Howard
CSHL
howard-at-cshl.org







From: Kalman Rubinson :      rubinsnk-at-is2.nyu.edu
Date: Thu, 10 Jul 1997 18:07:48 -0400 (EDT)
Subject: Re: Image Pro Plus

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On Thu, 10 Jul 1997 wise-at-vaxa.cis.uwosh.edu wrote:
} We have an older, DOS-based version of Image Pro Plus image
} analysis software that has us stymied. We are capturing video images of
} chloroplast movements and dumping them into a DOS-based computer. We want
} to be able to sum all of the white areas in a field and then express that
} total white area as a percentage of the total image area. It seems simple
} enough, but it's not. We can identify all of the "white" objects, we can
} define what we are calling 'white', we can get the area of each individual
} white object, but we cannot sum the areas of all of the white objects.
} Image Pro Plus corporation is no help. They sold the rights to the DOS
} version to another company and no one a IPP knows anything about it. They
} are quite willing to sell us the Windows version for $3500 but cannot help
} us with their older version.
} Does anyone out there in microscope land have experience with this
} program?

Yes, a bit. Is the remaining non-white area contiguous? If so, measure
that as the reciprocal of the white areas.

Kal





From: chandler-at-lamar.ColoState.EDU (John Chandler)
Date: Thu, 10 Jul 1997 16:18:53 -0600 (MDT)
Subject: TEM: Used Zeiss 902 Available

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I just heard of a Zeiss 902 TEM that is available for immediate purchase.

Location: USA
5 years old
400 hours of logged beam time
Top entry goniometer stage
C-mount port
EELS detector, but no computer

Contact: Tim Koons (voice) 847-658-3930 (FAX) 847-658-3993

PLEASE DO NOT REPLY TO THIS LIST WITH INQUIRIES. THANKS.

John
chandler-at-lamar.ColoState.EDU






From: psic-at-uclink4.berkeley.edu (Paula Sicurello)
Date: Thu, 10 Jul 1997 15:20:40 -0700 (PDT)
Subject: Source for Dissecting Scope?

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Hello All!


I'm trying to find a vendor who might sell a dissecting microscope
that has 2 heads. We need something like this for demonstration purposes.
It's a lot easier to use one of those than to keep switching chairs and
asking if they see what we're talking about.
So if anybody knows please send me the information so I can check
into it further.

Thanks!


Paula = )


Paula Sicurello
UC Berkeley
Electron Microscope Lab
psic-at-uclink4.berkeley.edu






From: bozzola-at-siu.edu (John J. Bozzola)
Date: Thu, 10 Jul 1997 16:56:54 -0600
Subject: Re: formvar background and pta/u.a. stains

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} I am currently staining milk proteins with phosphotungtic acid (2%/water
} ph 7.3) and uranyl acetate (1%/water) and often I do get what I called a
} background consisting of very small (about 20-40nm) bubbles-like
} structures that interfere with my samples...
}
} I also recall seeing the same thing with viruses but this time it is rather
} annoying because it also looks a lot like my protein...


This annoying phenomenon, sometimes called "champagning", may be caused by
several factors. In my own experience, I would see it whenever using PTA in
preparations with high protein concentrations. If you are quick, you can
actually see the phenomenon taking place as the beam vaporizes the PTA. I
believe that the proteins serve to sequester water which is then enrobed in
PTA. When the beam strikes the hydrated proteins, the water is vaporized
and bursts through the PTA crust (like erupting gases in magma) leaving
behind uniformly sized, spherical, electron light areas that may be
mistaken for viruses or protein subunits. One workaround may be to dilute
the protein, use phosphomolybdic or silicotungstic acids as the negative
stain and to dry the grids in a 60 degree oven over the weekend.


####################################################################
John J. Bozzola, Ph.D., Director
Center for Electron Microscopy
Neckers Building, Room 146 - B Wing
Southern Illinois University
Carbondale, IL 62901-4402
U.S.A.
Phone: 618-453-3730
Fax: 618-453-2665
Email: bozzola-at-siu.edu
Web: http://www.siu.edu/departments/shops/cem.html
####################################################################






From: maher-at-mat.mte.ncsu.edu (Dennis M. Maher)
Date: Thu, 10 Jul 1997 18:15:44 -0500
Subject: amorphous SiO2

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I am looking for a software package to construct and visualize an
amorphous 3-D SiO2 network with options that allow for insertion of other
elements such as nitrogen, carbon, and hydrogen and also the possibility of
forming various molecular groups such as silanol, siloxane, etc..

Does anyone know if such a software package exists?

Thanks,

David Wolfe (c/o Dr. D. Maher, NCSU)






From: buffat-at-cime.epfl.ch ( =?iso-8859-1?Q?Philippe=2DAndr=E9?= Buffat)
Date: Fri, 11 Jul 1997 07:40:47 +0100
Subject: Re: Electron diffraction patterns

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Sara wrote:

} I am looking for an electron diffraction analysis software package. I have
} to identify some particles (wether they're cubic, tetragonal, etc) and
} there must be an easier way than to hand draw the expected diffraction
} patterns....Does anybody know of something useful on the Web?

Try the EMS On-line soft running at WWW URL http://cimewww.epfl.ch/. This
is a short version of P. Stadelmann EMS programme.

Sara,
You can build a library of possible structures (lattice parameters, space
group and if known the atom position in the cell to account for kinematical
extinctions). Then you enter two or three diffracted vectors (lengths on
the pattern, angles, approximate camera length) and the programme tells you
on which zone axis of which structure you took the diffraction pattern plus
the exact camera length needed to do the fit. If there are some
ambiguities, you can also view/print the computed pattern=8A and more!

Philippe-A. Buffat


__________________________________________________________________
Philippe Buffat
Ecole Polytechnique Federale de Lausanne (EPFL)
Centre Interdepartemental de Microscopie Electronique
Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland
Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time)
E-mail: philippe.buffat-at-cime.uhd.epfl.ch, WWW URL http://cimewww.epfl.ch/
______________________________ Eudora F2.1 ___________________________






From: Eugen Schwan :      SCHWAN-at-hera.EMBL-Heidelberg.DE
Date: Fri, 11 Jul 1997 07:59:33 +0100
Subject: unsubscribe

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unsubscribe from the list






From: ifj. Dr. Kasa Peter :      KASA-at-pharma.szote.u-szeged.hu
Date: Fri, 11 Jul 1997 08:37:12 MET
Subject: Unsubscribe

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Please unsubscribe me from July 12 to Aug. 20.

Thanks

Peter





From: Jacky Larnould :      larnould-at-mnet.fr
Date: Fri, 11 Jul 1997 10:50:40 +0200
Subject: Re: History...Wolfram and Tungsten

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At 08:49 10/07/1997 -0600, you wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

Wolfram is a German word and it's a reference to the gray color of "Tungsten"
similar to the color of a wolf.
Tungsten is a swedish word and means heavy stone.
We use the word "Tunstene" in France, Wolfram is an alternate word but not
very common.
Salutations.
==========================================================
Jacky Larnould
mailto:larnould-at-mnet.fr
voice:33 (0)4 67 72 28 26
fax :33 (0)4 67 79 54 90




From: S.Suder-at-eee.salford.ac.uk
Date: 11 Jul 97 9:03
Subject: Diff.pattern for porous superlattice silicon?

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Dear all,

I came across some sort of porous superlattice silicon(I think) in my
rearch, I would be happy if someone provides me some information on
diffraction patterns of porous superlattice silicon.

Thanks in advance


S Suder



Suli Suder
Joule Physics Lab
University of Salford
Salford M5 4WT
United Kingdom
Tel: 0161 745 5000 ext. 53264
Fax: 0161 745 5119
E-mail: s.suder-at-eee.salford.ac.uk





From: Jim Darley :      jim-at-proscitech.com.au
Date: Fri, 11 Jul 1997 20:16:12 +1000
Subject: Re: Confocal list

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Mark et.al. -
We have the confocal listserver linked in our comprehensive listing of
microscopy sites. When you go to it you will get this notice:

"Season's Greetings
Our server is down for repairing.
We will gradually bring the web up in the coming week.
Please visit us after the New Year.
Have a happy holiday!"

http://corn.eng.buffalo.edu/www/ConfocalList/C1994/Q1/0037.html

I think they are a bit premature!
Seasons greetings
Jim Darley

ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313
Great microscopy catalogue, 400+ Links, MSDS
************************ http://www.proscitech.com.au

_________________________
} Dear all,
} Could someone please give me the address of the listserver for
} confocal microscopy.
}
} Thanks a lot,
}
} Mark Munro
} SAC Aberdeen
} m.munro-at-ab.sac.ac.uk




From: Jim Darley :      jim-at-proscitech.com.au
Date: Fri, 11 Jul 1997 20:16:12 +1000
Subject: Re: Confocal list

Contents Retrieved from Microscopy Listserver Archives
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Mark et.al. -
We have the confocal listserver linked in our comprehensive listing of
microscopy sites. When you go to it you will get this notice:

"Season's Greetings
Our server is down for repairing.
We will gradually bring the web up in the coming week.
Please visit us after the New Year.
Have a happy holiday!"

http://corn.eng.buffalo.edu/www/ConfocalList/C1994/Q1/0037.html

I think they are a bit premature!
Seasons greetings
Jim Darley

ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313
Great microscopy catalogue, 400+ Links, MSDS
************************ http://www.proscitech.com.au

_________________________
} Dear all,
} Could someone please give me the address of the listserver for
} confocal microscopy.
}
} Thanks a lot,
}
} Mark Munro
} SAC Aberdeen
} m.munro-at-ab.sac.ac.uk




From: Jim Darley :      jim-at-proscitech.com.au
Date: Fri, 11 Jul 1997 20:23:45 +1000
Subject: Re: Pyroxylin coating

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Jean and all: Here is a copy from our on-line catalogue:

"PARLODION Pyroxylin, in strips.
A highly purified form of cellulose nitrate; prepared for embedding tissu=
es
for sectioning and for making support films for EM grids from 1% parlodio=
n
in Amyl Acetate."

I prefer parlodion over formvar because it seems to release better when
cast on a microscope slide, also is seems more suitable for thicker film=
s.
It comes in hard strips. Cut and weigh a suitable piece and then calcula=
te
the solvent required to make the percentage solution. Because it is very
flammable parlodion it is frequently stored under water. Blot, then dry =
in
an incubator before weighing the small piece. Parlodion, like formvar
requires=20
a carbon coating for stability in TEM. Butvar does not.=20
ProSciTech and I trust all other EM suppliers handle Parlodion.
=20
Jim Darley
ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313
Great microscopy catalogue, 400+ Links, MSDS =20
************************ http://www.proscitech.com.au
=20
----------
} From: Leclerc Jean {leclercj-at-magellan.umontreal.ca}
} Hi there!=88
} =20
} Anyone knows anything about pyroxylin? I would like to know what it is
} exactly, and if it might be more stable than some other films for coati=
ng
} grids for the TEM. (Note that I'm looking at thick stuff - no thin
} sections here!)
} =20
} Thanks!
} ----------
} =20




From: ebs-at-ebsciences.com
Date: Fri, 11 Jul 1997 07:32:34 EST
Subject: faster resinless embedding

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Dear fellow microscopists,

At 10:49 AM 7/10/97 -0400, Michael Delannoy wrote:
I'm in the process of using the DGD resinless embedding technique developed
by } Capco, Krochmalnic and Penman and I'm looking for: 1)Short cuts-
} the ethanol, nbutanol to DGD transition is very long (6 hours) and I am
working } with monolayers

{snip}

It is often possible to dramatically speed up the dehydration steps of TEM
specimen preparation by carrying them out in a laboratory microwave. The
technique is described on pp352-353 of _The Microwave Cookbook for
Microscopists._

disclaimer: Energy Beam Sciences manufactures laboratory microwaves, and
has a vested interest in seeing more people utilizing this technique.

Best regards,
Steven E. Slap, Vice-President
********************************
Energy Beam Sciences, Inc.
Adding Brilliance To Your Vision
ebs-at-ebsciences.com
http://www.ebsciences.com/
********************************





From: Richard E. Edelmann :      edelmare-at-casmail.muohio.edu
Date: Fri, 11 Jul 1997 07:45:31 -0500
Subject: Sticky Grid Box Puzzler

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O.K. you professionals here's a real puzzler for you:


I have a user here who has about 24 SPI Slide-A Grid Boxes.
They were a little dusty (they may have been used before - heavens
not to say they didn't come absolutely pristine from SPI), and so he
throughly washed them and rinsed them with Ethanol, only now he can
not get them back open any longer! They are really stuck.

I don't know if he removed some type of lubication on the box,
maybe the "natural" lube from the manufacturing process, or if he
caused the plastic to swell with the EtOH, but I could certianly use
any suggestion. Maybe we should throw this up to the group - surely
someone has cleaned grib boxes before.

We can get the box lids to move upto 5mm or so but then they stop
and with work we can get them to move back, but we still have freed
any lids. We've tried prying up the lids a little from the back
edges (where they slide along rasied ridges) - didn't help much.
We've even tried soaking them again in water or EtOH hoping to
provide some "lubrication" (there are no grids in the boxes, but
didn't really want to soak them in some thing oily) - still with no
success.


I oringinally asked this of Charles Garber (of SPI) and neither of us
have come up with any ideas, but he did throw in the following:

Of the box parts:

} The plastic is an antistatic formulation however, alcohol being so
} polar is not going to swell the base piece at least. If the alcohol
} was hot or if it was in contact with the plastic for some extended
} period of time, then who knows, but I just can not believe that the
} alcohol would have done anything to it in the way you are
} suggesting.
}
} The sliding top might be another story. I forgot the plastic that is
} used for it, but it might be a clear PVC. But even then, room
} temperature alcohol should not have had time to do anything.
}

Any suggestions?

The things fools will do ....


Richard E. Edelmann, Ph.D.
Electron Microscopy Facility Supervisor
352 Pearson Hall
Miami University, Oxford, OH 45056
Ph: 513-529-5712 Fax: 513-529-4243
E-mail: edelmare-at-muohio.edu


"640K ought to be enough for anybody."
-- Bill Gates, 1981




From: hoopea01-at-mchip00.med.nyu.edu (Andrea Therese Hooper)
Date: Fri, 11 Jul 1997 08:39:26 -0500
Subject: Re: confocal list

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I believe that this is the Confocal list address:

{LISTSERV-at-LISTSERV.ACSU.BUFFALO.EDU}

Andrea T. Hooper
NYU Medical Center






From: Bo Johansen :      Boj-at-bot.ku.dk
Date: Fri, 11 Jul 1997 08:47:12 -0700
Subject: Re: faster resinless embedding

Contents Retrieved from Microscopy Listserver Archives
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ebs-at-ebsciences.com wrote:
} It is often possible to dramatically speed up the dehydration steps of TEM
} specimen preparation by carrying them out in a laboratory microwave. The
} technique is described on pp352-353 of _The Microwave Cookbook for
} Microscopists._

Another way to speed up dehydration is to do it chemically - use
acidified 2,2 dimethoxypropane (DMP). For a monlayer complete
dehydration will occur in a few min. I usually dehydrate small blocks
(up to 3x3x3 mm) of plant tissue for 15 min, and then go directly to
embedding medium:dimethoxypropane 1:1. Any residual water from the
dehydration step will be removed during the first embedding step.
However, I do not know if DGD can be mixed or react with DMP, so you
have to test it first.


Bo





From: Cheri Owen :      cowen-at-cmb.biosci.wayne.edu
Date: Fri, 11 Jul 1997 09:33:19 -0400 (EDT)
Subject: Tungsten shadowing

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Does anyone know a TEM protocol for "Tungsten Shadowing" say, for DNA
strand enhancement?

Thanks!

Cheri Owen
Detroit Neurotrauma Institute
Wayne State University
Detroit





From: Cheri Owen :      cowen-at-cmb.biosci.wayne.edu
Date: Fri, 11 Jul 1997 09:43:24 -0400 (EDT)
Subject: Re: TEM Vibration Isolation

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Hi Brad,

Any reason why you wouldn't use an anti-vibration platform? We have a
Micro-g isolation table made by Technical Manufacturing Corporation for
our JEM 1010. We were in a room next to the furnace and air conditioners
for the whole building and NEVER had any problems. If you are interested,
I have the information, so should JEOL, since they both have offices in
Peabody, Massachusetts.

Cheri Owen
Detroit Neurotrauma Institute
Wayne State University
Detroit





From: dave :      dave.strecker-at-ab.com
Date: Fri, 11 Jul 1997 09:49:13 -0400
Subject: M&M97 early registration deadline is approaching!

Contents Retrieved from Microscopy Listserver Archives
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Microscopy & Microanalysis '97 in Cleveland is less than a month away!
The deadline for early registration is this coming Tuesday, July 15. If
you have not yet done so, register now to qualify for the lower rates.
If you need registration forms, please call the business office at
(800) 538-3672 . If you have internet access to the www, forms are
available for downloading at
http://www.bright.net/~strecker/msno/mm97.html.

If you are planning on attending and have not yet reserved hotel space,
you must contact the hotels directly. Hotel information is also
available from our web site or from the business office. I can also
e-mail you this information if required. At our last planning meeting
it was announced that there are no rooms available downtown Cleveland
for Saturday, August 9, so if you need a room, please act now.

A special note to those who have been following the Mars Pathfinder
program. A debate on the pros and cons of life on Mars will take place
on Monday, August 11. Following this will be a presentation by Al
Worden, Apollo 15 astronaut. Mr. Worden's talk will feature many of his
breath-taking photographs from space.

----------------------------------------------------------------
Dave Strecker mailto:dave.strecker-at-ab.com
Rockwell Automation/Allen-Bradley Phone: (216)646-3250
Component Engineering ND246 Fax: (216)646-3416
1 Allen-Bradley Dr.
Mayfield Hts., Ohio 44124 USA
----------------------------------------------------------------






From: Conrad Perfett :      ConradPerfett-at-pasttimes.com
Date: Fri, 11 Jul 1997 14:55:34 -0000
Subject: M&M97 early registration deadline is approaching!

Contents Retrieved from Microscopy Listserver Archives
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Sorry Geoff! I guess I was forgetting that other people may want to know
the details of this book, so I am posting the details here:

A beginners guide to the collection, isolation, cultivation and
identification of Freshwater Protozoa

Pblished in 1988 by
Culture Collection of Algae Associaction (CCAP)

Freshwater Biological Association
The Ferry House
Ambleside
Cumbria
United Kingdom

ISBN 1 871105 03 X

but remember it has drawings not photos! still for 4.40 uk pounds
you can't complain :)

} -----Original Message-----
} From: Geoff McAuliffe [SMTP:mcauliff-at-UMDNJ.EDU]
} Sent: Friday, July 11, 1997 4:34 PM
} To: Conrad Perfett
} Subject: protozoa book??
}
} Dear Conrad:
}
} You have been on the MSA list praising a book on microscopy of protozoa
} quite often. So what is the title of the book??????? I keep reading your
} posts hoping to find some mention of the title, author, publisher,
} something! Either post the title or email it to me, please.
}
} 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 e-mail: mcauliff-at-umdnj.edu
} ***************************************************************




From: Robert Underwood :      underwoo-at-u.washington.edu
Date: Fri, 11 Jul 1997 07:20:33 -0700 (PDT)
Subject: Re: fluorescence?

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Greetings,
I received many replies to my original query, and in case others
are interested, here is the gist of them. Thanks to all of you who answered.


Hi Tamara,

If you have a proper filter setup, the emission filter should be excluding
all the exitation wavelength. Therefore I think you are looking at
autofluorescence. Any light that has not changed wavelength should be
blocked.

Bob

On Thu, 10 Jul 1997, Tamara Howard wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Disclaimer first: I'm not a chemistry/physics person, so please
} excuse me if this question is incredibly stupid :)
} Is it possible for a pigment to scatter/reflect light strongly
} enough to look like fluorescence? If so, how could you distinguish such
} behaviour from "real" fluorescence? I'm trying to work with some pigmented
} plant samples that "glow" under my rhodamine/Tx Red excitation/emission
} filters (standard fluorescence scope); the pigmented areas look normal
} under FITC and UV conditions. The pigment itself is red under regular
} light. We'd like to know if the pigment autofluoresces or if this is just
} some light/pigment interaction. I'm lost and our library isn't heavy on
} this type of references. Any of you "hard science" types willing to try to
} educate me?!
}
} TIA!
}
} Tamara Howard
} CSHL
} howard-at-cshl.org
}
}
}
}





From: Ulf Skoglund :      ulf.skoglund-at-cmb.ki.se
Date: Fri, 11 Jul 1997 17:00:44 +0200
Subject: unsubscribe

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unsubscribe





From: Mike Coviello :      Coviello-at-mae.uta.edu
Date: Fri, 11 Jul 1997 10:00:29 -0500
Subject: Enlargers-LogEtronics EM-55

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Hello Everyone:

Does anyone know where I can get a used LogEtronics 55-EM enlarger
(or something similar with auto-dodge & burn-in capabilities)in good
working condition?


Thanks,

Michael Coviello
Lab Manager
The University of Texas -at- Arlington




From: MICHAEL DELANNOY :      delannoy-at-welchlink.welch.jhu.edu
Date: Fri, 11 Jul 1997 11:54:57 -0400 (EDT)
Subject: Resinless embedding

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Hello again,
DGD stands for diethylene glycol disterarate, which is a
waxy "solid" at room temp (although sweats/melts at r.t. like the London
resins-white,gold) and liquid at 70 deg.C. The references are:
Pennman (1995) PNAS. 92:5251-5257 review
Capco etal (1984) JCB. 98:1878-1885
These will get you started.
Right now I've run some samples on tissue culture plates, coverslips
and a glass slide (which got trashed). Separating the DGD from the plates
was easy (light hammer tapping) but some cells did remain on the dish.
The coverslips separated easily, so I've mounted a few and will be cutting
soon (I can't see the cells so I'm sectioning blindly). One problem is the
block sweating at the cell surface, I've placed everything in the fridge
hopefully to stop this-cutting should be interesting. The second paper
used glass petris which I don't have, but may have to get. The n-butanol
did not dissolve the culture plate. I'll keep you posted, more suggesstions
are welcome.

Sweating in Baltimore,
Mike D.

P.S. I got my DGD from Polysciences who also sent a protocol. This stuff
may work better with tissue blocks and pellets, monolayers are tricky.




From: rtind-at-siu.edu (Nadia Navarrete)
Date: Fri, 11 Jul 1997 11:29:30 -0600
Subject: Enlargers

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Michael,

An excellent source for used photographic equipment of all kinds, including
digital imaging, is a monthly magazine called "Shutterbug". I think it's
put out by the same people who do the "Computer Shopper" magazine. They
began as an advertising magazine dedicated to used photo stuff, but have
expanded into a large format glossy, general photography magazine. They
still have loads of ads and classifieds for all sorts of used you-name-it
relating to photography. It can be found in many camera shops and magazine
newstands.

Randy Tindall
Center for Electron Microscopy
Southern Illinois University at Carbondale






From: HILDEGARD CROWLEY :      hcrowley-at-du.edu
Date: Fri, 11 Jul 1997 10:31:31 -0600 (MDT)
Subject: Re: Sticky grid boxes

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Hi,
In 25 years of being in a microscopy laboratory I have witnessed impossibly
sticky grid boxes 3 times. Twice they were manufactured by LKB, once they came
from another source. Each time the grid boxes were washed in alcohol.
No time were we able to rescue the boxes. We threw them away. We now
strictly wash all grid boxes with soap and water only.
So sorry,
HHC





From: Scott D. Ireland :      sireland-at-frontiernet.net
Date: Fri, 11 Jul 1997 10:55:52 -0400
Subject: Re: Image Pro Plus

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Bob -

In Image-Pro for DOS, after you count and measure all the white objects,
the total area of these objects can be found by going to View-Statistics.
The Sum measurement is the total area of all counted objects.

Additionally, to clarify a few points, Image-Pro for DOS was not sold to
anyone - Media Cybernetics - the original developer of the Image-Pro family
of products - still owns all rights to these products. However, the DOS
versions of Image-Pro were discontinued quite some time ago (nearly 4
years). Although we make our best effort to support all of our products
and customers, there have been at least 6 new Windows versions published
since the last DOS version, and the vast majority of our customers have
taken advantage of the upgrades or extended maintenance plans in order to
stay 'current'. If you have registered your product with us, then you have
probably seen the announcements and upgrade offers over the years.

If you, or any other DOS users out there, would like to upgrade to the
latest version - Image-Pro Plus 3.0, please feel free to contact me
directly.

--------------------------------------
Scott D. Ireland
Regional Sales Manager
Eastern North America & Latin America
Media Cybernetics, LP
"The Imaging Experts"
Tel: (716) 473-0222
Fax: (716) 473-8048
scott-at-mediacy.com
http://www.mediacy.com
---------------------------------------

----------
} From: wise-at-vaxa.cis.uwosh.edu
} To: Microscopy-at-sparc5.microscopy.com
} Subject: Image Pro Plus
} Date: Thursday, July 10, 1997 9:21 AM
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} To all,
}
} We have an older, DOS-based version of Image Pro Plus image
} analysis software that has us stymied. We are capturing video images of
} chloroplast movements and dumping them into a DOS-based computer. We
want
} to be able to sum all of the white areas in a field and then express that
} total white area as a percentage of the total image area. It seems
simple
} enough, but it's not. We can identify all of the "white" objects, we can
} define what we are calling 'white', we can get the area of each
individual
} white object, but we cannot sum the areas of all of the white objects.
} Image Pro Plus corporation is no help. They sold the rights to the DOS
} version to another company and no one a IPP knows anything about it.
They
} are quite willing to sell us the Windows version for $3500 but cannot
help
} us with their older version.
} Does anyone out there in microscope land have experience with
this
} program?
}
} Yours in computer frustration,
}
} Bob
}
}




From: ejb11-at-psu.edu (Edward J. Basgall)
Date: Fri, 11 Jul 1997 13:47:10 -0500
Subject: Re: Tungsten shadowing

Contents Retrieved from Microscopy Listserver Archives
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}
} Does anyone know a TEM protocol for "Tungsten Shadowing" say, for DNA
} strand enhancement?
}
} Thanks!
}
} Cheri Owen
} Detroit Neurotrauma Institute
} Wayne State University
} Detroit

Cheri,
There are two sections on tungsten shadowing in Chap 7 " High Resolution
Shadowing" by Henry S. Slayter in the book: Electron Microscopy in
Biology, J.R. Harris, ed. 1991, IRL Press at Oxford Univ. Press. ISBN
0-19-963215-4 (pbk)

cheers
ed

Edward J. Basgall, PhD
The Pennsylvania State University
Surface Chemistry Group ejb11-at-psu.edu
Materials Research Institute Building Ph: 814-865-0493
University Park, PA 16802-7003 FAX: 814-863-0618






From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Fri, 11 Jul 1997 15:33:32 -0500
Subject: RUSH PRINTS! Rush Lab.

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Often in our lab we are in a big rush to get prints right after the user
has finished using the EM. I've sometimes just washed the negatives
(3.25X4" Kodak EM film) just 2 minutes instead of 20, and then after
I've printed them all, I go back and wash them for 30 minutes to remove
any trace fixative that might still be remaining in the emulsion. This
way I can shave 18 minutes off of the delivery time. I also print them
with just a bit of water at their edges so that they are not completely
dry, and shave another 5 minutes off of the drying time, so that they
are 23 minutes out the door faster.

Does anyone else have any long term experience with this wash-after-wash
technique. I'd like to know more about the long term effect, i.e.:
whether I can indeed wash them later and be sure to remove all the
traces of fix from the film emulsion.

In a hurry,
Garry
}




From: Kalman Rubinson :      rubinsnk-at-is2.nyu.edu
Date: Fri, 11 Jul 1997 16:31:14 -0400 (EDT)
Subject: Re: Image Pro Plus

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On Fri, 11 Jul 1997, Scott D. Ireland wrote:

} However, the DOS
} versions of Image-Pro were discontinued quite some time ago (nearly 4
} years). Although we make our best effort to support all of our products
} and customers, there have been at least 6 new Windows versions published
} since the last DOS version, and the vast majority of our customers have
} taken advantage of the upgrades or extended maintenance plans in order to
} stay 'current'.

There is a difference in perspective about these issues for many of us.

'Nearly 4 years' is not a long time for the life of a useful product after
discontinuation and, in fact, I am often loath to upgrade software if the
current version is working despite the 'enhancements' of newer versions.
For example, previous correspondance has revealed that IP has changed some
of the ways in which it measures certain parameters and I am concerned
about data compatibility. In addition, interface changes are tedious even
if the newer interface is better; one has to unlearn too many things.

So, your statement that IPWin has had at least 6 versions since the last
DOS version (which I have) is an indication of progress but it is also a
matter of great concern about support for these lapsed versions.

Kalman Rubinson
NYU Medical Center





From: William Tivol :      tivol-at-wadsworth.org
Date: Fri, 11 Jul 1997 17:26:41 -0500 (EDT)
Subject: Re: RUSH PRINTS! Rush Lab.

Contents Retrieved from Microscopy Listserver Archives
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} Often in our lab we are in a big rush to get prints right after the user
} has finished using the EM. I've sometimes just washed the negatives
} (3.25X4" Kodak EM film) just 2 minutes instead of 20, and then after
} I've printed them all, I go back and wash them for 30 minutes to remove
} any trace fixative that might still be remaining in the emulsion. This
} way I can shave 18 minutes off of the delivery time. I also print them
} with just a bit of water at their edges so that they are not completely
} dry, and shave another 5 minutes off of the drying time, so that they
} are 23 minutes out the door faster.
}
} Does anyone else have any long term experience with this wash-after-wash
} technique. I'd like to know more about the long term effect, i.e.:
} whether I can indeed wash them later and be sure to remove all the
} traces of fix from the film emulsion.
}
Dear Garry,
We don't use the wash-after-wash technique; rather, we wash for
~1 min in H2O, 1 min in Permawash and ~1 min in H20. This removes the
fixer and negs which have been processed this way keep for several
years at least. I guess if a user were *really* in a hurry, we could
save ~1 min, but our way the user can take the negs home. ;-)
Yours,
Bill Tivol




From: Larry Allard :      allardlfjr-at-ornl.gov
Date: Fri, 11 Jul 1997 21:25:22 -0400
Subject: Re: RUSH PRINTS! Rush Lab.

Contents Retrieved from Microscopy Listserver Archives
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A *very* good argument for Digital Imaging!

Larry


}
} } Often in our lab we are in a big rush to get prints right after the user
} } has finished using the EM. I've sometimes just washed the negatives
} } (3.25X4" Kodak EM film) just 2 minutes instead of 20, and then after
} } I've printed them all, I go back and wash them for 30 minutes to remove
} } any trace fixative that might still be remaining in the emulsion. This
} } way I can shave 18 minutes off of the delivery time. I also print them
} } with just a bit of water at their edges so that they are not completely
} } dry, and shave another 5 minutes off of the drying time, so that they
} } are 23 minutes out the door faster.
} }
} } Does anyone else have any long term experience with this wash-after-wash
} } technique. I'd like to know more about the long term effect, i.e.:
} } whether I can indeed wash them later and be sure to remove all the
} } traces of fix from the film emulsion.
} }
} Dear Garry,
} We don't use the wash-after-wash technique; rather, we wash for
} ~1 min in H2O, 1 min in Permawash and ~1 min in H20. This removes the
} fixer and negs which have been processed this way keep for several
} years at least. I guess if a user were *really* in a hurry, we could
} save ~1 min, but our way the user can take the negs home. ;-)
} Yours,
} Bill Tivol


Dr. Lawrence F. Allard
Senior Research Staff Member
High Temperature Materials Laboratory
Oak Ridge National Laboratory
1 Bethel Valley Road
Bldg. 4515, MS 6064
PO Box 2008
Oak Ridge, TN 37831-6064

423-574-4981
423-574-4913 Fax
l2a-at-ornl.gov






From: T.T. COZZIKA FOUNDATION :      cozzika-at-compulink.gr
Date: Sat, 12 Jul 1997 11:39:04 -0500
Subject: leukocytes - electron microscopy

Contents Retrieved from Microscopy Listserver Archives
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Please recommend, if it's possible, suitable, detailed references or
your own procedure, how to isolate leukocyte-rich plasma and process it
without sedimentation media, for electron microscopy.

Thanks for your understanding,
Margarita Chrysanthou
E-mail : cozzika-at-athena.copmulink.gr






From: Beth Bray :      bbray-at-netside.com
Date: Sat, 12 Jul 1997 14:05:36 -0400
Subject: Ignore

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

Sorry. Please ignore the "subscribe" request you just received from me as I
am already subscribed to the list.

Beth Bray




From: Gabriel Adriano Rosa :      micros-at-biolo.bg.fcen.uba.ar
Date: Sat, 12 Jul 1997 17:00:14
Subject: Looking for a TEM and-or SEM

Contents Retrieved from Microscopy Listserver Archives
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The Department of Biological Sciences of the Exacts and Natural Sciences
Faculty
of the Buenos Aires University are looking for donation of a TEM and-or SEM
equipment,
not older than 15 years and still woorking.
We will paid any handling and shipping cost.
It would be very helpfull for us any other kind of assistance.
Thank you vey much in advance.

Please respond to me at the addresses and numbers listed below.




Gabriel Adriano Rosa
Area Microscopia Electronica, Depto. Cs. Biologicas
Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires
Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA

FAX (54-1)-782-0582 e-mail micros-at-biolo.bg.fcen.uba.ar




From: Jim Darley :      jim-at-proscitech.com.au
Date: Sat, 12 Jul 1997 17:45:23 +1000
Subject: Re: Tungsten shadowing

Contents Retrieved from Microscopy Listserver Archives
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Cheri and all -
Specific to DNA strands, I know this as the Kleinschmitt technique, now
over 25 years old. The osmotically opened DNA strands are rotary shadowed.
For that the specimen grids are rotated at about 60 rpm, 60-100mm from the
source and at an 6-10 degree angle of incidence. The rotary shadowing makes
it easier to check the continuity and to follow the under/over of the
fibers.
I used to evaporate Pt/Pd or for finer grain, Pt and C simultaneously.
Ed Basgall's posting should help with the tungsten evaporation. Tungsten is
finer still but really requires special equipment. It is possible to
evaporate W - "Wolfram" in a normal evaporator, but it is slow and a lot
of heat is generated. A heat shield for the specimen with a suitable
aperture is advised.
But why that trouble? The coarser grain from other evaporation material
does not impair tracing and measuring of the DNA fibres and fibre detail is
better using negative staining, or positive staining using methanolic UA.
Regards
Jim Darley

ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313
Great microscopy catalogue, 400+ Links, MSDS
************************ http://www.proscitech.com.au

----------
} From: Cheri Owen {cowen-at-cmb.biosci.wayne.edu}
} To: Microscopy-at-sparc5.microscopy.com
} Subject: Tungsten shadowing
} Date: Friday, 11 July 1997 23:33
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
}
} Does anyone know a TEM protocol for "Tungsten Shadowing" say, for DNA
} strand enhancement?
}
} Thanks!
}
} Cheri Owen
} Detroit Neurotrauma Institute
} Wayne State University
} Detroit
}




From: nkrsmith-at-juno.com (Nancy K. R. Smith)
Date: Sat, 12 Jul 1997 19:26:12 -0500
Subject: AFM position, San Antonio

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A Research Associate position is open in the dDpartment of Radiology,
University of Texas Health Science Center at San Antonio, as the primary
technician for the operation of a new atomic force
microscope (Nanoscope, Digital Instruments). The person in this position
would participate in studies of novel vascular biomaterial surfaces as
well as in studies of vascular cell responses to mechanical forces.
Applicants should have a Master's degree or a bachelor's degree with
several years experience . Preferred areas of training and/or experience
include scanning electron microscopy, image analysis, and cell biology.
The position is currently open and we seek to hire someone as soon as
possible. All applicants must apply through Human Resources, UTHSCSA.
Inquiries should be directed to Dr. Gene Sprague, 210 567-5564, e-mail
sprague-at-uthscsa.edu





From: Peter Michiels :      pmi-at-mail2.tornado.be
Date: Mon, 14 Jul 1997 17:11:51 +0100
Subject: unsubscribe

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Please unsubscribe me.
___________________________________

Ing. Peter Michiels

Philips Analytical - Electron Optics
Tel.: +32-2-5256249
Fax: +32-2-5256483
e-mail: pmi-at-tornado.be
___________________________________





From: Ian MacLaren :      I.MacLaren-at-BHAM.AC.UK
Date: Mon, 14 Jul 1997 16:37:08 +0100
Subject: Re: Suggestions on the twin-jet electropolisher

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Ibrahim and anyone else interested,

We have used the Struers Tenupols here for many years and are generally
happy with them. They wear out after a while because of all the nasty
electropolishing solutions that people want to use in them but they seem to
be good for at least 3-5 years (probably a lot longer if they did't get the
battering that our students and RFs give them). Even then, the first
things to go are the jets and these can be replaced (at a price).

You do (at least with tenupols) have to think a bit about how you will cool
your electrolytes. We have a system where alcohol cooled with either dry
ice or liquid nitrogen is pumped through the cooling coils on the tenupol.
The temperature of the electrolyte is controlled (to give between 0 and -40
degrees C) by a homebuilt control box using the reading from a electrical
resistance thermometer to control a relay that turns the pumping of the
cooling alcohol on or off.

Hope this helps


++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Ian MacLaren, Tel: (44) (0) 121 414 3447
IRC in Materials for FAX: (44) (0) 121 414 3441
High Performance Applications, email: I.MacLaren-at-bham.ac.uk
The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/
Birmingham B15 2TT,
England.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++






From: dana nojima :      noji-at-lenti.med.umn.edu
Date: Mon, 14 Jul 1997 09:38:49
Subject: RE: RUSH PRINTS! Rush Lab.

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When I had worked as a newspaper photographer back in my college days, time
was always something that we were trying to cheat on. We used the EtOH
wash technique to get things to dry faster.

One thing that has not been addressed is what are the photos going to be
used for. Previously a typical morning at the EM would produce 80 to 100
films. We would print 8X10 images of these photos. These would later be
used to keep track of the films and to keep track of the portions that
would were later digitized. The printing to RC paper and processing with
an Ilford developer went fairly quickly. I would hate to think of how
much time it would take to send 100 several meg images to the Fugi
Pixtograph printer. First there would be the storage problem, the output
problem, lower resolution, and then the cost.

Dana
noji-at-snowman.med.umn.edu __~0_\___
Minneapolis, Minnesota (_________)
(612) 624-4687 O O
http://www.tc.umn.edu/nlhome/g118/post-doc/dtn.htm






From: MICHAEL DELANNOY :      delannoy-at-welchlink.welch.jhu.edu
Date: Mon, 14 Jul 1997 11:49:11 -0400 (EDT)
Subject: 488 nm excitable autofluorescence

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Microscopists,
Are there any new tricks for depleting autofluorescence (488 nm
excit) in fixed culture cells? Thanks.

Mike D.




From: Robert Mixon :      mixonr-at-ohsu.edu
Date: Mon, 14 Jul 1997 08:52:11 -0700
Subject: Leukocytes-reply

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Margarita: One can simply make a buffy coat by spinning the sample in a
wintrobe tube ( a skinny glass tube obtained from hematology) immediately and
fixing the buffy coat by aspirating off most of the serum and replacing with
glutaraldehyde/paraformaldehyde EM fixative. then cut the top and bottom off
the wintrobe tube (after diamond scoring) and float this half inch long segment
in the fixative until the buffy starts to get firm. Simply take a wooden
applicator stick and poke the pellet out of the tube segment, process for EM and
embed on edge to see all the leukocyte types in layers. bob M




From: Microscopy-request
Date: Monday, July 14, 1997 8:33AM
Subject: Suggestions on the twin-jet electropolisher

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Ibrahim:

We have a Fishione unit which we have used for the past fifteen years. We
found the unit to be very reliable. Our unit came with a glass dish used as
the reservoir. This tended to affect the response of the light detector
used to stop the electropolishing when a hole appeared. In most instances
however this was not a severe limitation for us. The jets and the specimen
holder do need to be aligned occasionally. Also, we did have to replace the
jets and the holder once since the electrodes do have a finite life. As
long as the unit is rinsed thoroughly after use it should last a long time.
We found the unit to be very compact and it was a simple matter to cool the
electrolyte using a double beaker with the larger beaker containing the
coolant. Please not that I am not familiar with the newer units and that
configuration might have changed by now.

I have no financial interests in Fischione.

Jordi Marti




----------
-----------------------------------------------------------------------.


Hi everyone,

I need your suggestions on the twin-jet electropolishers for
preparation of TEM samples. I need a reliable brand with reasonable
price. Right now, I have information from Struers,SouthBay Technology and
Fischione. Fischione has a reasonable price. Does anyone have any
experience with Fischione?

Thank you for your all suggestions. Please reply my own adress.

Ibrahim Karaman
Mechanical Engineering Department
University of Illinois
1206 W. Green Street
Urbana, IL 61801





From: RWILLSON-at-pearl.tufts.edu
Date: Mon, 14 Jul 1997 13:30:03 -0500 (EST)
Subject: unsubscribe

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unsubscribe




From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Mon, 14 Jul 1997 12:51:51 -0500
Subject: Permawash

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Bill,

I've never heard about Permawash. I've heard of "hypoclear" from Kodak,
but the instructions state that this should be discarded after 24 hours,
so it would be expensive and time consuming to make this up all the
time. Is this Permawash stuff stable enough that I could keep it in a
stainless steel covered tank for a week or so, to use as required?

Garry



} } Dear Garry,
} } We don't use the wash-after-wash technique; rather, we wash for
} } ~1 min in H2O, 1 min in Permawash and ~1 min in H20. This removes the
} } fixer and negs which have been processed this way keep for several
} } years at least. I guess if a user were *really* in a hurry, we could
} } save ~1 min, but our way the user can take the negs home. ;-)
} } Yours,
} } Bill Tivol
}
}




From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Mon, 14 Jul 1997 12:55:14 -0500
Subject: RE: Rush Lab

Contents Retrieved from Microscopy Listserver Archives
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Randy,

It would have been my thoughts that the film need not be washed for such
a long period of time, but I'm just trying to follow the instructions as
supplied by Kodak as a film insert. Somehow they felt that 20 minutes
was the minimum time required unless one used hypoclear or Permawash.

Garry

} Garry,
}
} It's probably not necessary to use the wash-after-wash technique. William
} Tivol's reply probably provides the best compromise. Film, as opposed to a
} print on paper, is not really very absorbent and can be washed pretty clean
} pretty fast. Fix tends to rinse out readily. (I repeat, this is NOT
} necessarily true for resin-coated prints, although they also wash pretty
} fast, and is CERTAINLY not true for fiber-based prints.)
}
} As an example, in their IlfoPro newsletter, Ilford recently recommended a
} wash technique for their 35mm and roll films that involved filling the
} developing tank with water and inverting it 5 times, refilling it and
} inverting it 10 times, and filling it a third time and inverting it 20
} times. They claim this is an archival wash technique.
}
} After only a couple minutes wash, I'd bet that your negatives will show no
} signs of change for many years. (I can already hear the screams of
} outrage!) Hypo clearing agents, PermaWash, etc., are excellent for
} assuring that the negatives will outlive the people taking them, and I
} certainly recommend their use for safety's sake. I still have negatives,
} however, that were taken when I was 12 years old and processed in a
} makeshift darkroom (it required nighttime in the country to make it
} light-tight, IF the moon wasn't too bright). They were washed pretty
} sloppily and still show no signs of deterioration 30 years later.
}
} Regarding digital imaging---in my opinion, silver-based photography is
} still the most cost-effective, highest information content, easily stored
} method for image CAPTURE. After capture, however, digital is more and more
} the way to go for manipulation and publication. Nothing yet beats a
} negative as an imaging starting point, especially considering the sizeable
} investment in decent digital imaging equipment. Most labs already have
} darkrooms. (I bet this will generate some comments!)
}
} Hope this helps.
}
} Randy Tindall
} Center for Electron Microscopy
} Southern Illinois University at Carbondale
}
}
}




From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Mon, 14 Jul 1997 13:00:11 -0500
Subject: Rush Lab. -Why?

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Brian,

We are doing diagnostic electron microscopy as part of the Department of
Pathology, in a major health care centre. The users that demand fast
turn around time are the Pathologists especially when we are processing
a fine needle aspirate biopsy sample. Our technique spares the patient
from more invasive procedures. Because of our speed, we have been able
to deliver the micrographs within 8 working hours from fresh tissue, to
dispell the myth that EM as applied to biological tissue is necessarily
very very slow. Often we get micrographs after 6 working hours or less.
So, speed is of the essence for us. (at least that is what they tell
us)

Garry

} ----------
} From: Brian G. Demczyk[SMTP:demczyk-at-erxindy.rl.plh.af.mil]
} Sent: 13 July, 1997 06:35
} To: Garry Burgess
} Subject: Re: RUSH PRINTS! Rush Lab.
}
} Just out of curiosity, what industry are you in that users
} demand such a rapid turn around?!
}




From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Mon, 14 Jul 1997 14:02:28 -0500
Subject: RE: Rush Lab + Projection Slides

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Randy,

Sometimes when I want to make a transparency for seminars, or workshops
or whatever, I put the EM negative into the enlarger and project it to a
small 2X2" size on to yet another piece of EM film that I've cut in half
for this purpose. (you need a special enlarger lens to get this small!).
Then I cut it to size and mount it in a glass slide mount, and voila, I
have supersize black and white slides, with good resolution and
contrast. (at least better than 35mm projection slides) But sometimes
in the past, if I tried to rush things, I notice that the image turned
brown. To fix this problem though, I simply re-fix and re-wash this
image, and the brown discoloration (which is probably some residual
silver halide and fix) is removed, and all is well again.

Garry

} ----------
} From: rtind-at-siu.edu[SMTP:rtind-at-siu.edu]
} Sent: 14 July, 1997 14:51
} To: Garry Burgess
} Subject: RE: Rush Lab
}
} } ------------------------------------------------------------------------
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America




From: bmh7-at-cornell.edu
Date: Mon, 14 Jul 1997 16:30:44 -0400 (EDT)
Subject: Re: Permawash

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from what i understand, Permawash is just soapy water. i simply use a
mild detergent solution instead and it works fine

} I've never heard about Permawash. I've heard of "hypoclear" from Kodak,
} but the instructions state that this should be discarded after 24 hours,
} so it would be expensive and time consuming to make this up all the
} time. Is this Permawash stuff stable enough that I could keep it in a
} stainless steel covered tank for a week or so, to use as required?

} } } We don't use the wash-after-wash technique; rather, we wash for
} } } ~1 min in H2O, 1 min in Permawash and ~1 min in H20. This removes the
} } } fixer and negs which have been processed this way keep for several
} } } years at least. I guess if a user were *really* in a hurry, we could
} } } save ~1 min, but our way the user can take the negs home. ;-)




From: David Knecht :      knecht-at-uconnvm.uconn.edu
Date: Mon, 14 Jul 1997 15:54:34 -0500
Subject: ConA-labeled

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Does anyone know any good sources besides Sigma for conA labeled with
Rhodamine, Texas Red or another non-FITC dye. Thanks- Dave

Dr. David Knecht
Department of Molecular and Cell Biology
University of Connecticut
U-125
Storrs, CT 06269
Knecht-at-uconnvm.uconn.edu






From: Buddy Steffens :      steffens-at-calc.vet.uga.edu
Date: Mon, 14 Jul 1997 17:20:32 EST
Subject: Re: L.M. "Evans Blue" stain?

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Evans blue is commonly used as a counterstain for flourescence work.
It stains the specimen for observation by white light, and since it
has no autoflourescence, it is invisible under the UV wavelengths.

For this application, a bottle should last an academic lifetime...I
can't imagine what a case would be used for (except maybe spending
surplus year-end funds)




-=W.L. Steffens=-
Department of Veterinary Pathology
College of Veterinary Medicine
University of Georgia
Athens, GA 30602 USA
http://www.vet.uga.edu/vpp/wls/steffens.html




From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Mon, 14 Jul 1997 16:26:03 -0500
Subject: RE: RUSH PRINTS! Rush Lab.

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Dana,

The photos are going to be used by Pathologists in a hospital for
diagnostic purposes.

Garry
}
} One thing that has not been addressed is what are the photos going to be
} used for. Previously a typical morning at the EM would produce 80 to 100
} films. We would print 8X10 images of these photos. These would later be
} used to keep track of the films and to keep track of the portions that
} would were later digitized. The printing to RC paper and processing with
} an Ilford developer went fairly quickly. I would hate to think of how
} much time it would take to send 100 several meg images to the Fugi
} Pixtograph printer. First there would be the storage problem, the output
} problem, lower resolution, and then the cost.
}
} Dana
} noji-at-snowman.med.umn.edu __~0_\___
} Minneapolis, Minnesota (_________)
} (612) 624-4687 O O
} http://www.tc.umn.edu/nlhome/g118/post-doc/dtn.htm
}
}
}




From: William Tivol :      tivol-at-wadsworth.org
Date: Mon, 14 Jul 1997 17:50:01 -0500 (EDT)
Subject: Re: Permawash

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} I've never heard about Permawash. I've heard of "hypoclear" from Kodak,
} but the instructions state that this should be discarded after 24 hours,
} so it would be expensive and time consuming to make this up all the
} time. Is this Permawash stuff stable enough that I could keep it in a
} stainless steel covered tank for a week or so, to use as required?
}
Dear Garry,
The bottle of Permawash says that working solution will oxidise
in an open tank/tray in 6-8 hrs. A tank with a floating lid will keep
for 30 days and a stoppered bottle for 90 days. The undiluted stock
solution will keep a lot longer.
We have our working solution in a SS covered tank like yours;
I'm confident that it keeps easily for the week or so you need.
Yours,
Bill Tivol




From: js_vetrano-at-ccmail.pnl.gov (John S Vetrano)
Date: Mon, 14 Jul 1997 14:26:33 -0700
Subject: Inexpensive LM revisited

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Greetings, all;

Sorry to do this to you, but there was a discussion a while back about
inexpensive light microscopes for children. In particular there was a
recommendation from someone in the Seattle area about a $99 model (Tasco
brand?). Of course, I deleted the messages but was recently asked by a friend
for a recommendation. I just about went blind looking (unsuccesfully) through
the June archive so it must have been further back then that! To save my eyes
from further abuse, could the person posting about that microscope please just
e-mail me the company name and address? I would greatly appreciate it.

Oh, also any recommendations for things to look at for kids ages 6-10 would be
appreciated.

Thanks in advance.

Cheers,

John Vetrano
Pacific Northwest National Laboratory
P.O. Box 999
Richland, WA 99352

js_vetrano-at-pnl.gov




From: Evex EDS :      evex-at-pluto.njcc.com
Date: Sun, 13 Jul 1997 14:25:08 -0400
Subject: www.evex.com update

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Our Home Page has been updated





From: ibrahim karaman :      karaman-at-students.uiuc.edu
Date: Mon, 14 Jul 1997 08:33:31 -0500 (CDT)
Subject: Suggestions on the twin-jet electropolisher

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Hi everyone,

I need your suggestions on the twin-jet electropolishers for
preparation of TEM samples. I need a reliable brand with reasonable
price. Right now, I have information from Struers,SouthBay Technology and
Fischione. Fischione has a reasonable price. Does anyone have any
experience with Fischione?

Thank you for your all suggestions. Please reply my own adress.

Ibrahim Karaman
Mechanical Engineering Department
University of Illinois
1206 W. Green Street
Urbana, IL 61801





From: ad408-at-detroit.freenet.org (Gilbert T. Groehn)
Date: Sun, 13 Jul 1997 11:20:21 -0400
Subject: >>L.M. "Evans Blue" stain?<<

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Our lab acquired a case of EVANS BLUE stain
from one of our clients and we can not locate
this in any of our chemistry manuals.

What are the normal uses for this stain (eg.
histology, hematology, etc.) ? We would be
using it only for light microscopy.

What is the standard mixing ratios and
formulas for various applications?

Any help on this item would be most appreciated.

TIA
Gil Groehn
ULTRAMED, INC.

--
=============================================================
ULTRAMED, INC. Research Div. ad408-at-detroit.freenet.org
Grosse Pointe Farms, MICH 48236 USA Biomedical Consultants
===============================================================




From: csedax-at-alpha.arcride.edu.ar
Date: Mon, 14 Jul 1997 11:29:53 -2359
Subject: SEM: sample preparation for particles dispersed in oil???

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,
Dear friends from the list,


We have just received an enquiry about SEM analysis
for: a) pigments particles dispersed in mineral oils
b) pigments particles in oil-water emulsions

Does any of you have any experience in that
subject? Unfortunately, we do not count with equipment for sample preparation
other than the sputter-coater and carbon evaporator.

Any help will be very welcome.

Thanks in advance,

Nora Pratta
Centro Regional de Investigacion y Desarrollo
Santa Fe - Argentina









From: Neelima Shah :      shahn-at-mail.med.upenn.edu
Date: Sat, 12 Jul 1997 10:41:21 -0400
Subject: Re: RUSH PRINTS! Rush Lab.

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}
} A *very* good argument for Digital Imaging!
}
} Larry

Yes! but at what cost! with dwindling funds and tighter budgets all
arguments are in vain.........
Neelima Shah
Regards...
:-) :-) :-) :-) :-) :-) :-) :-) :-) :-) ;-) ;-)
Visit us at http://www.med.upenn.edu/~path/core/EMCMAIN1.htm





From: John W Heckman :      heckman-at-pilot.msu.edu
Date: Mon, 14 Jul 1997 08:45:39 -0400 (EDT)
Subject: Re: Staining Plants for Phenolics

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----------------snip---------------}
} 3] Is there any fixative other than Osmium which would best retain phenolics
} in small root explants?
}
} I am not using a cryoprotectant like DMSO because I want to avoid soaking the

} roots in it, as this could allow phenolics to diffuse from the plant cells.
} However, small root explants are soaked in Os-KI for several hours under
} vacuum, to overnight at 4 C prior to microtomy. This allows for Os
} penetration. I am relying on Os to "fix" the phenolics. Perhaps there is
} something else which would penetrate more rapidly and fix or condense the
} phenolics.
}
} Mahalo = Hawaiian for Thanks !!!!!!!!
}

Dave,

have you tried RuO4? I have found it works well with a number of unsaturated
ring structure compounds as a fixative. I make it up from the chloride just
before use and it seems to act quite rapidly.

cheers,
John




From: AMCGroup2-at-aol.com
Date: Mon, 14 Jul 1997 03:09:14 -0400 (EDT)
Subject: TEM Specimen Prep Course

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Advanced TEM Specimen Prep Techniques Overview: A Short Course

* Precision Angle Lapping (PAL); the =93Wedge Technique=94
* Focused Ion-Beam (FIB) Milling w. and w/o. Mechanical Pre-thinning
* Materials Ultramicrotomy (MUM)
* Precision Angle Cleaving (PAC)

This new one-day course provides an intensive overview of all the above
techniques. The program mainly focuses on the practical aspects of the
methods, the criteria for their selection, the latest developments in the=
ir
applications and also reviews the related instrumentation, tools, and
accessories.

The next 1997 upcoming course will be offered on August 15th in Cleveland=
,
OH, following the MSA Annual Meeting. During the rest of this year, the
course will be offered eight more times in U.S., Europe, and Asia. Pleas=
e
note that our regular 3-day hands-on workshops covering each one of the a=
bove
techniques are still offered twice a year in May and November.

To receive a copy of the short course brochure including the Registration
Form and detailed information, please provide us with your complete maili=
ng
address, fax, and phone numbers. Thank you.

Rene E. Nicholas
AMC Group
(A Division of Promotech Associates, Inc.)
e-Mail amcgroup2-at-aol.com







From: Stephan Coetzee :      stephan-at-gecko.biol.wits.ac.za
Date: Mon, 14 Jul 1997 15:37:54 GMT+2
Subject: Re: Amoeba in Nature

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Dear Conrad
}
} } well I now know why I am having difficulty finding the Amoeba since
} } according to the book I have just bought, at least the classic Amoeba
} } Proteus is rare in the wild!!! I remember being told by a retired
} } microscopist that it was rare also.
}
I did send a previous reply to the list. Since I got no copy I have
to assume it is lost? Depending on what you see as "wild", "nature"
and "natural", Amoeba is easy to find in large concentrations. At
water refinery plants they are in high concentrations in the
activated sludge. There is normally a buildup of material on the
edge above water level, different species and even "waterbears" can
be found. (Remember to use gloves collecting and handling samples.)

##
[########]
##
##
##
##
##
Stephan H Coeztee
Electron Microscope Unit
Private Bag 3
Wits
2050
South Africa

Stephan-at-Gecko.biol.WITS.ac.za

Tell: +27 11 716 2419
Fax : +27 11 339 3407

}
##
[########]
##
##
##
##
##
Stephan H Coeztee
Electron Microscope Unit
Private Bag 3
Wits
2050
South Africa

Stephan-at-Gecko.biol.WITS.ac.za

Tell: +27 11 716 2419
Fax : +27 11 339 3407




From: Garry Burgess
Date: 12 July 1997 00:47
Subject: RUSH PRINTS! Rush Lab.

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Garry

If time is of the essence have you considered doing a final wash in ethanol
(70% or thereabouts)?
It must be clean of course but you should knock a few more drying minutes
off. Try it on something unimportant first though.

Malcolm Haswell
e.m. unit
University of Sunderland
UK
----------

Often in our lab we are in a big rush to get prints right after the user has
finished using the EM. I've sometimes just washed the negatives (3.25X4"
Kodak EM film) just 2 minutes instead of 20, and then after
I've printed them all, I go back and wash them for 30 minutes to remove any
trace fixative that might still be remaining in the emulsion. This way I
can shave 18 minutes off of the delivery time. I also print them
with just a bit of water at their edges so that they are not completely dry,
and shave another 5 minutes off of the drying time, so that they are 23
minutes out the door faster.

Does anyone else have any long term experience with this wash-after-wash
technique. I'd like to know more about the long term effect, i.e.: whether
I can indeed wash them later and be sure to remove all the
traces of fix from the film emulsion.

In a hurry,
Garry





From: David Webb :      davehawaiiedu-at-msn.com
Date: Sat, 12 Jul 97 15:08:36 UT
Subject: Staining Plants for Phenolics

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We have been using two methods (Fe Acetate & Osmium KI) to stain for phenolics
in thick (30-60u) CRYOsections of plant roots. We checked these methods on
coffee leaves which have a lot of phenolics, and while both worked, the Os-KI
method was superior and more specific in terms of the literature. In our
research specimens (roots of alfalfa) neither of these works very well by
itself. However, we get a strong staining reaction from Os-KI treated
specimens if we add Toluidine Blue just prior to observation. This reveals
cells with large dark deposits in their vacuoles. These could be phenolics but
they could be something else. We are planning to screen several more
protocols, as it would be encouraging to have several methods which yield
approximately the same results, because the specificity of histochemical
methods is often not understood. I have the following questions.

1] Has anyone used Toluidine Blue in consort with other treatments (especially
Os-KI) to localize phenolics? In other words is this staining reaction
indicative of the presence of phenolics?

2] Are there any other histochemical procedures which are known to be specific
for phenolics that may be appropriate for frozen plant sections?


3] Is there any fixative other than Osmium which would best retain phenolics
in small root explants?

I am not using a cryoprotectant like DMSO because I want to avoid soaking the
roots in it, as this could allow phenolics to diffuse from the plant cells.
However, small root explants are soaked in Os-KI for several hours under
vacuum, to overnight at 4 C prior to microtomy. This allows for Os
penetration. I am relying on Os to "fix" the phenolics. Perhaps there is
something else which would penetrate more rapidly and fix or condense the
phenolics.

Mahalo = Hawaiian for Thanks !!!!!!!!




From: David Webb :      davehawaiiedu-at-msn.com
Date: Sat, 12 Jul 97 15:08:36 UT
Subject: Staining Plants for Phenolics

Contents Retrieved from Microscopy Listserver Archives
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We have been using two methods (Fe Acetate & Osmium KI) to stain for phenolics
in thick (30-60u) CRYOsections of plant roots. We checked these methods on
coffee leaves which have a lot of phenolics, and while both worked, the Os-KI
method was superior and more specific in terms of the literature. In our
research specimens (roots of alfalfa) neither of these works very well by
itself. However, we get a strong staining reaction from Os-KI treated
specimens if we add Toluidine Blue just prior to observation. This reveals
cells with large dark deposits in their vacuoles. These could be phenolics but
they could be something else. We are planning to screen several more
protocols, as it would be encouraging to have several methods which yield
approximately the same results, because the specificity of histochemical
methods is often not understood. I have the following questions.

1] Has anyone used Toluidine Blue in consort with other treatments (especially
Os-KI) to localize phenolics? In other words is this staining reaction
indicative of the presence of phenolics?

2] Are there any other histochemical procedures which are known to be specific
for phenolics that may be appropriate for frozen plant sections?


3] Is there any fixative other than Osmium which would best retain phenolics
in small root explants?

I am not using a cryoprotectant like DMSO because I want to avoid soaking the
roots in it, as this could allow phenolics to diffuse from the plant cells.
However, small root explants are soaked in Os-KI for several hours under
vacuum, to overnight at 4 C prior to microtomy. This allows for Os
penetration. I am relying on Os to "fix" the phenolics. Perhaps there is
something else which would penetrate more rapidly and fix or condense the
phenolics.

Mahalo = Hawaiian for Thanks !!!!!!!!




From: rtind-at-siu.edu (Randy Tindall)
Date: Sun, 13 Jul 1997 16:28:00 -0600
Subject: Re: Rush Lab

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Garry,

It's probably not necessary to use the wash-after-wash technique. William
Tivol's reply probably provides the best compromise. Film, as opposed to a
print on paper, is not really very absorbent and can be washed pretty clean
pretty fast. Fix tends to rinse out readily. (I repeat, this is NOT
necessarily true for resin-coated prints, although they also wash pretty
fast, and is CERTAINLY not true for fiber-based prints.)

As an example, in their IlfoPro newsletter, Ilford recently recommended a
wash technique for their 35mm and roll films that involved filling the
developing tank with water and inverting it 5 times, refilling it and
inverting it 10 times, and filling it a third time and inverting it 20
times. They claim this is an archival wash technique.

After only a couple minutes wash, I'd bet that your negatives will show no
signs of change for many years. (I can already hear the screams of
outrage!) Hypo clearing agents, PermaWash, etc., are excellent for
assuring that the negatives will outlive the people taking them, and I
certainly recommend their use for safety's sake. I still have negatives,
however, that were taken when I was 12 years old and processed in a
makeshift darkroom (it required nighttime in the country to make it
light-tight, IF the moon wasn't too bright). They were washed pretty
sloppily and still show no signs of deterioration 30 years later.

Regarding digital imaging---in my opinion, silver-based photography is
still the most cost-effective, highest information content, easily stored
method for image CAPTURE. After capture, however, digital is more and more
the way to go for manipulation and publication. Nothing yet beats a
negative as an imaging starting point, especially considering the sizeable
investment in decent digital imaging equipment. Most labs already have
darkrooms. (I bet this will generate some comments!)

Hope this helps.

Randy Tindall
Center for Electron Microscopy
Southern Illinois University at Carbondale






From: A. Kent Christensen :      akc-at-umich.edu
Date: Mon, 14 Jul 1997 10:49:09 -0400 (EDT)
Subject: Re: >>L.M. "Evans Blue" stain?<<

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Gil,

Herbert McLean Evans was chair of Anatomy at Univ of Calif Berkeley, from
1915. His early research included use of vital dyes for studies on blood
volume, macrophages, ovarian estrous cycle, etc. In a 1920 paper (Dawson,
Evans, Whipple. "Blood volume studies. III. Behavior of large series of
dyes introduced into the circulating blood." Amer J Physiol 51:232-256)
it was shown that a blue azo dye (T-1824) was "slightly superior" for
blood volume measurement than the vital red series previously used. The
dye came to be know as "Evans blue." Criteria that were important were:
(1) non-toxic, (2) not stored in tissue, (3) color allows accurate
determination, (4) removed quite slowly from the blood stream.

An example of more recent use: Bergh, Damber, 1988, Int J Androl 11:449.

Kent
(A. Kent Christensen, Dept of Anatomy and Cell Biology, University of
Michigan Medical School), {akc-at-umich.edu} )

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

On Sun, 13 Jul 1997, Gilbert T. Groehn wrote:

} Our lab acquired a case of EVANS BLUE stain
} from one of our clients and we can not locate
} this in any of our chemistry manuals.
}
} What are the normal uses for this stain (eg.
} histology, hematology, etc.) ? We would be
} using it only for light microscopy.
}
} What is the standard mixing ratios and
} formulas for various applications?
}
} Any help on this item would be most appreciated.
}
} TIA
} Gil Groehn
} ULTRAMED, INC.
}
} =============================================================
} ULTRAMED, INC. Research Div. ad408-at-detroit.freenet.org
} Grosse Pointe Farms, MICH 48236 USA Biomedical Consultants
} ===============================================================
}





From: Bennett, Cynthia, HDG / FHF :      bennett-at-msmhdg.hoechst.com
Date: Mon, 14 Jul 1997 09:46:00 +0200
Subject: AW: EDX - Spot Mode

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Stephen,

We use the spot mode pretty routinely to identify inorganic particles in
our polyester films. The size of these particles is anywhere from about
0.5 =B5m to 10 =B5m. So far it has worked quite well for us. When we =
switch
back to full scan after using the spot mode, we invariably see a "burn"
mark which is exactly where we think it ought to be. We often look at a
2nd spot away from the structure that interests us to be sure that we're
not just picking up a background.

Cindy Bennett
Hoechst Diafoil GmbH
Wiesbaden, Germany
bennett-at-msmhdg.hoechst.com




} I travel the world teaching practical electron microscopy and it =
worries me
} the emphasis that people place on "Spot Mode". I do not teach the use =
of
} spot mode for the following reasons-

} 1. Just because the spot appears in a certain position on the =
screen
} is this its correct position. I have found machines many microns out =
of
} step in X and Y directions.

} 2. Specimens charge and switching out of spot mode does not give =
the
} operator an easy opportunity to see if the spot had moved during the
} analysis.

} 3. Spot mode gives a false sense of analytical volume, no matter =
how
} small that spot may be we are almost certainly evaluating microns of
} material.

} Do others worry about spot mode accuracy, do others test the spot mode
} accuracy? Is it not better to simply increase the magnification =
watching
} the area of interest all the way up before the analysis and all the way
} down after the analysis?

} What do you think?




From: Nelissen, B.J. :      Bart.B.J.Nelissen-at-Arnhem.ACR.akzo.nl
Date: Mon, 14 Jul 1997 12:09:58 +0200
Subject: Re: TEM vibration isolation

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Brad,



Youre vibration problem resembles a problem I had about 7 years ago when
I was working with scanning tunneling microscopy at the university of
Nijmegen (The Netherlands). Our STM in a UHV system was on the 4th floor,
so we had some trouble with vibrations, mainly stemming from air
ventilation compressors in the base of the building. In the same basement
there were three vibration free sites consisting of concrete bloks (2x2x2
m) placed in sand an having no connection to the foundation of the
building. We measured the vibrations on these bloks and found that they
were only a little bit better than on the 4th floor!! We found out that
the vibrations from the compressors propagates not only through the
building construction but also through the ground or sand. So the effect
of de-coupling the concrete blocks from the building foundation did not
eliminate the vibration-coupling, but it did reduce the (effective) mass
of the blocks. Hence the vibration amplitude growths! The foundation of
the building, although directly coupled to the floors of the compressors,
showed a quit clean vibration spectrum, because the mass is high
(effectively the whole building).



So, back to your question: cutting the floor around your microscope will
only help if this floor is the only vibration coupling to the microscope.
If also the ground (dirt) or air (acoustic) couples vibrations to your
microscope, the end result will be worse, since the mass of your system
is reduced.



Success, Bart





Bart Nelissen

Akzo Nobel Central Research

Dept of Applied Physics, Microscopy

Tel: +31 26 366 1371

Fax: +31 26 366 5272

eMail: Bart.B.J.Nelissen-at-Akzo.nl



Note: Do not use automatic reply or answer facilities in your e-mail
program

to respond to this message (the sender-address is distorted by our
'gate-keeper').




From: edelmare
Date: 11 July 1997 14:26
Subject: Sticky Grid Box Puzzler

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I remember many years ago trying to clean some grid boxes (the white LKB
type with a clear lid) with ethanol and it all went disastrously wrong. I
can't remember whether both types of plastic deteriorated but the boxes were
unusable. It may be that SPI boxes are made of similar material - try asking
them.
I now just give boxes a wash with soapy water (maybe a quick ultrasonic
clean) and rinse in distilled water then dry for a couple of days
I don't know if this helps.

Malcolm Haswell
e.m. unit
University of Sunderland
UK
----------

O.K. you professionals here's a real puzzler for you:

I have a user here who has about 24 SPI Slide-A Grid Boxes.
They were a little dusty (they may have been used before - heavens
not to say they didn't come absolutely pristine from SPI), and so he
throughly washed them and rinsed them with Ethanol, only now he can
not get them back open any longer! They are really stuck.

I don't know if he removed some type of lubication on the box,
maybe the "natural" lube from the manufacturing process, or if he
caused the plastic to swell with the EtOH, but I could certianly use
any suggestion. Maybe we should throw this up to the group - surely
someone has cleaned grib boxes before.

We can get the box lids to move upto 5mm or so but then they stop
and with work we can get them to move back, but we still have freed
any lids. We've tried prying up the lids a little from the back
edges (where they slide along rasied ridges) - didn't help much.
We've even tried soaking them again in water or EtOH hoping to
provide some "lubrication" (there are no grids in the boxes, but
didn't really want to soak them in some thing oily) - still with no
success.


I oringinally asked this of Charles Garber (of SPI) and neither of us
have come up with any ideas, but he did throw in the following:

Of the box parts:

} The plastic is an antistatic formulation however, alcohol being so
} polar is not going to swell the base piece at least. If the alcohol
} was hot or if it was in contact with the plastic for some extended
} period of time, then who knows, but I just can not believe that the
} alcohol would have done anything to it in the way you are
} suggesting.
}
} The sliding top might be another story. I forgot the plastic that is
} used for it, but it might be a clear PVC. But even then, room
} temperature alcohol should not have had time to do anything.
}

Any suggestions?

The things fools will do ....

Richard E. Edelmann, Ph.D.
Electron Microscopy Facility Supervisor
352 Pearson Hall
Miami University, Oxford, OH 45056
Ph: 513-529-5712 Fax: 513-529-4243
E-mail: edelmare-at-muohio.edu


"640K ought to be enough for anybody."
-- Bill Gates, 1981





From: chris gilpin :      cgilpin-at-fs1.sem.man.ac.uk
Date: Wed, 9 Jul 1997 16:27:58 BST
Subject: Re: PHOSPORWOLFRAM ACID

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}
} Can any of our German colleagues help with translation please? Came
} across the term PHOSPHORWOLFRAM acid in a technical paper. Suspect it is
} phosphoric acid but want to be sure before proceeding.
} Thanks in advance
} Ronnie Houston
I think you will find that it is phosphotungstic acid (hence the
atomic symbol W for tungsten)

Chris


Chris Gilpin
Biological Sciences Electron Microscope Unit
G452 Stopford Building
Oxford Road
Manchester
M13 9PT
phone +44 161 275 5170
fax +44 161 275 5171
http://www.biomed.man.ac.uk/biology/emunit/emhome.html




From: Karen L. Vaughn :      klv-at-biotech.ufl.edu
Date: Tue, 15 Jul 1997 09:31:34 -0400
Subject: neg stain of virus

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I have a client who will be purifying a retrovirus using a sucrose gradient.

The question is what medium should the sample be in to do the negative
stain. Sucrose? Buffer? If anyone has worked with retrovirus I would like
to hear your suggestion.

Thank you in advance

Karen Vaughn,EM Technician phone: (352)392-1184
University of Florida fax: (352)846-0251
Electron Microscopy Core Lab. e-mail: klv-at-biotech.ufl.edu
214 Bartram Hall web:
http://www.biotech.ufl.edu/~emcl/
Gainesville, Fl. 32611






From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Tue, 15 Jul 1997 09:51:37 -0500
Subject: RE: Rush Lab + Projection Slides

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Garry,

I'm not sure what is causing the brown staining, especially since you
indicate that it's temporary. My GUESS is that it's inadequate fixing
time
or exhausted fixer. What are your fixing times?

Randy,

Usually I fix for 2-3 minutes, but in this case, it might be more, since
the slides were actually just piled up in a staining beaker filled with
hypo. Since the slides were in contact with each other, I'm quite sure
that this explains the inadequate fixation. Actually usually I look at
them under the safelight to ensure that they are adequately fixed, but
sometimes, when I had a huge volume, then I must have missed a couple,
because I had so many. (about 200 or so)

I use some sort of Kodak fixer, but I can't remember the name right at
this second. I usually do not use hypochek, because as I've said,
usually it's obvious when the fixer is losing it's strength, because it
starts to take significantly longer than 2 minutes to fix our EM
negatives. Because we work under safelight, this is something that can
be checked each and every time that we develop film.


What fix do you use? Do
you check your fix for exhaustion with HypoChek or a similar product
that
gives you a white precipitate when the fixer is saturated with silver
compounds? Interesting.

I agree with you about the confusion about Permawash. It would have to
be something more chemically like Hypo Clear than soap, because I can't
imagine a soapy solution removing fix from the emulsion, though it's an
interesting thought. However, I also don't think that I'd like a soapy
solution as a substitute for Photoflo, because I think that a soapy
solution might leave a residue, which is probably the last think that
one would want in the last step of their film developing.

Incidentally, someone on the server said that they thought PermaWash was
a
soap solution. I think they're confusing it with PhotoFlo, which, as
you
know, is just a wetting agent to prevent water drops from marking the
film
when it dries. PermaWash is more like Hypo Clear in that it chemically
removes fixer or changes it into more soluble, harmless compounds (I
forget
which!). Another very good product is Orbit Bath, which is very cheap
and
effective, but unfortunately seems to be now sold under a different
name.
Ask in a good photo shop, if you're interested. There is also Hypo
Eliminator, another Kodak product, which the Kodak folks once told me is
different than Hypo Clear and, I believe, is meant strictly for films,
rather than both film and paper.

The history of these compounds seems to be kind of interesting, because
I've read somewhere that ordinary sea water works as a hypo clearing
agent
and that it forms the basis for the modern products. Take it for what
it's
worth...

Neat method of making slides, by the way. I'm going to remember it for
possible future use.

Yes, it works fine. But the biggest obstacle for someone doing this for
the first time is to make sure that they have a lens that is capable of
making such a small focused image. You also have to make sure that you
use glass slide holders, because negative film cannot stand up to the
heat of a slide projector without glass protection, because it's not as
tough as slide film.

Garry



} ----------
} From: rtind-at-siu.edu[SMTP:rtind-at-siu.edu]
} Sent: 15 July, 1997 09:08
} To: Garry Burgess
} Subject: RE: Rush Lab + Projection Slides
}
} Garry,
}
} I'm not sure what is causing the brown staining, especially since you
} indicate that it's temporary. My GUESS is that it's inadequate fixing time
} or exhausted fixer. What are your fixing times?
}
} Randy,
}
} Usually I fix for 2-3 minutes, but in this case, it might be more, since the
} slides were actually just piled up in a staining beaker filled with hypo.
} Since the slides were in contact with each other, I'm quite sure that this
} explains the inadequate fixation. Actually usually I look at them under the
} safelight to ensure that they are adequately fixed, but sometimes, when I had
} a huge volume, then I must have missed a couple, because I had so many.
} (about 200 or so)
}
} I use some sort of Kodak fixer, but I can't remember the name right at this
} second. I usually do not use hypochek, because as I've said, usually it's
} obvious when the fixer is losing it's strength, because it starts to take
} significantly longer than 2 minutes to fix our EM negatives. Because we work
} under safelight, this is something that can be checked each and every time
} that we develop film.
}
}
} What fix do you use? Do
} you check your fix for exhaustion with HypoChek or a similar product that
} gives you a white precipitate when the fixer is saturated with silver
} compounds? Interesting.
}
} I agree with you about the confusion about Permawash. It would have to be
} something more chemically like Hypo Clear than soap, because I can't imagine
} a soapy solution removing fix from the emulsion, though it's an interesting
} thought. However, I also don't think that I'd like a soapy solution as a
} substitute for Photoflo, because I think that a soapy solution might leave a
} residue, which is probably the last think that one would want in the last
} step of their film developing.
}
} Incidentally, someone on the server said that they thought PermaWash was a
} soap solution. I think they're confusing it with PhotoFlo, which, as you
} know, is just a wetting agent to prevent water drops from marking the film
} when it dries. PermaWash is more like Hypo Clear in that it chemically
} removes fixer or changes it into more soluble, harmless compounds (I forget
} which!). Another very good product is Orbit Bath, which is very cheap and
} effective, but unfortunately seems to be now sold under a different name.
} Ask in a good photo shop, if you're interested. There is also Hypo
} Eliminator, another Kodak product, which the Kodak folks once told me is
} different than Hypo Clear and, I believe, is meant strictly for films,
} rather than both film and paper.
}
} The history of these compounds seems to be kind of interesting, because
} I've read somewhere that ordinary sea water works as a hypo clearing agent
} and that it forms the basis for the modern products. Take it for what it's
} worth...
}
} Neat method of making slides, by the way. I'm going to remember it for
} possible future use.
}
} Yes, it works fine. But the biggest obstacle for someone doing this for the
} first time is to make sure that they have a lens that is capable of making
} such a small focused image. You also have to make sure that you use glass
} slide holders, because negative film cannot stand up to the heat of a slide
} projector without glass protection, because it's not as tough as slide film.
}
} Garry
}
}




From: Caroline Schooley :      schooley-at-mcn.org
Date: Tue, 15 Jul 1997 08:02:29 -0700
Subject: Re: Inexpensive LM revisited

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} Sorry to do this to you, but there was a discussion a while back about
} inexpensive light microscopes for children. In particular there was a
} recommendation from someone in the Seattle area about a $99 model (Tasco
} brand?). Of course, I deleted the messages but was recently asked by a friend
} for a recommendation.

Younger children do MUCH better with an erect image, so the first decision
is "dissecting" vs. compound. And another choice is monocular vs.
binocular. Young children have problems with both the eye spacing and the
convergance required by binocs, so the extra cost of two oculars may not be
justified.
}
} Oh, also any recommendations for things to look at for kids ages 6-10 would be
} appreciated.

Children want to DO things, rather than just look. My grandson was
fascinated with watching epsom salt crystals grow when he was 6. You'll
find a wealth of suggestions in the Project MICRO bibliography (see below);
look in section IIA, "The microscopic world".



Caroline Schooley
Educational Outreach Coordinator
Microscopy Society of America
Box 117, 45301 Caspar Point Road
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html
Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/






From: Sara Miller :      saram-at-acpub.duke.edu
Date: Tue, 15 Jul 1997 11:12:58 -0400 (EDT)
Subject: Re: ConA-labeled

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On Mon, 14 Jul 1997, David Knecht wrote:

} Date: Mon, 14 Jul 1997 15:54:34 -0500
} From: David Knecht {knecht-at-uconnvm.uconn.edu}
} To: microscopy-at-sparc5.microscopy.com
} Subject: ConA-labeled
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Does anyone know any good sources besides Sigma for conA labeled with
} Rhodamine, Texas Red or another non-FITC dye. Thanks- Dave
}
} Dr. David Knecht
} Department of Molecular and Cell Biology
} University of Connecticut
} U-125
} Storrs, CT 06269
} Knecht-at-uconnvm.uconn.edu
}
Linscott's Directory of Immunological and Biological Reagents (ISBN:
0-9604920-4-6, Phone: 415-544 9555, about $70) is a fantastic reference
that lists almost every antibody made by every company. It is a valuable
book I recommend for anyone doing immunostaining.}

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: ad408-at-detroit.freenet.org (Gilbert T. Groehn)
Date: Tue, 15 Jul 1997 11:24:12 -0400
Subject: Evans Blue Stain

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Thank you profusely to all of the members of the group who
responded to my request for info on Evans Blue Stain.

Your help is most appreciated.

Cordially,
Gil Groehn
ULTRAMED, INC.

--
=============================================================
ULTRAMED, INC. Research Div. ad408-at-detroit.freenet.org
Grosse Pointe Farms, MICH 48236 USA Biomedical Consultants
===============================================================




From: wesleysm-at-biology.und.ac.za (James Wesley-Smith)
Date: Tue, 15 Jul 1997 08:56:57 +-200
Subject: L.M. "Evans Blue" stain

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Hello Gilbert and cyber-folk
Evans blue can also be used to determine cellular integrity in plant cell
suspensions, squashes, etc. The dye will not penetrate intact plasmalemma,
while staining damaged cells.

Yet another use for your case of stain! Have you considered a "garage
sale"?


James Wesley-Smith
EM Unit
University of Natal
Durban, South Africa

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

On Sun, 13 Jul 1997, Gilbert T. Groehn wrote:

} Our lab acquired a case of EVANS BLUE stain
} from one of our clients and we can not locate
} this in any of our chemistry manuals.
}
} What are the normal uses for this stain (eg.
} histology, hematology, etc.) ? We would be
} using it only for light microscopy.
}
} What is the standard mixing ratios and
} formulas for various applications?
}
} Any help on this item would be most appreciated.
}
} TIA
} Gil Groehn
} ULTRAMED, INC.
}
} =============================================================
} ULTRAMED, INC. Research Div. ad408-at-detroit.freenet.org
} Grosse Pointe Farms, MICH 48236 USA Biomedical Consultants
} ===============================================================
}








From: Damian_Neuberger_at_RLT013-at-ccmailgw.mcgawpark.baxter.com
Date: 7/11/97 10:00 AM
Subject: Enlargers-LogEtronics EM-55

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Mike,

It would be interesting to know a situation where this enlarger would be the
preferred method of producing prints as opposed to taking the negatives that
need dodge/burn, scan them and then digitally "correct" them? Good luck on
finding this enlarger used etc,!

Damian Neuberger
neuberd-at-baxter.com

______________________________ Reply Separator _________________________________


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Hello Everyone:

Does anyone know where I can get a used LogEtronics 55-EM enlarger
(or something similar with auto-dodge & burn-in capabilities)in good
working condition?


Thanks,

Michael Coviello
Lab Manager
The University of Texas -at- Arlington
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From: bozzola-at-siu.edu (John J. Bozzola)
Date: Tue, 15 Jul 1997 13:36:14 -0600
Subject: Re: Permawash

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} from what i understand, Permawash is just soapy water. i simply use a
} mild detergent solution instead and it works fine
}
You may be thinking of Photoflo - which is essentially a wetting agent, a
detergent - used to break the hydrophobicity of film and permit sheeting
of the water. This is similar to the wetting agents used in dishwashing
machines (e.g., for spotless glassess). Permawash should contain some
chemical scavengers used to remove resisual fixers in the film/papers. Any
photo-chemical types listening to this?


####################################################################
John J. Bozzola, Ph.D., Director
Center for Electron Microscopy
Neckers Building, Room 146 - B Wing
Southern Illinois University
Carbondale, IL 62901-4402
U.S.A.
Phone: 618-453-3730
Fax: 618-453-2665
Email: bozzola-at-siu.edu
Web: http://www.siu.edu/departments/shops/cem.html
####################################################################






From: Steve Chapman :      PROTRAIN-at-CompuServe.COM
Date: Tue, 15 Jul 1997 15:35:14 -0400
Subject: EDX - Spot Mode and more on SEM

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I am interested in the size of particle that you talk about, what densit=
y
do you see in your material having carried out the analysis?

For materials similar to carbon the volume of material involved with an
electron beam of an energy suitable for most EDX analysis (say 15kV) wil=
l
be in excess of 2.7 microns? To carry out an analysis of 0.5 micron
structures with material the density of carbon would require a beam energ=
y
of less than 6kV or for iron less than 10kV.

The mark you call a "burn" is this not really contamination? There is
usually no damage to a specimen if you see a dark mark, contamination. T=
he
problem is that with your materials you may well drive off the volatile
components and then you will burn out an area of specimen, the depth
dependant upon the time of your observation or analysis.

However you seem to have conclusive proof that you are analysing the area=

that you have made your target. My point was not that a spot analysis is=

taboo but that we should not blindly use a spot analysis without knowing
more about its accuracy. Some replies have taken the attitude that becau=
se
they have used the technique for many many years there cannot be a proble=
m!
Unless you know exactly what is happening in a microscope I do not agree=

that this attitude is sufficient, I can show you instruments where this
attitude would be a disaster.

In the consultancy business the biggest problem that one confronts is the=

"scientist" who has been doing things his way for 20 years and will not
change. I understand that we may well compare driving a microscope to
driving a car. Who would by a new car and then go on a course to learn t=
o
drive it? The same approach in SEM however would be constructive; new
instruments open up new techniques and new areas of investigation. From
what I see the topic of scanning electron microscopy changes considerably=

within a two year cycle and operators who persist in using their 20 year
old techniques, even on an old instrument, are somewhat lost! The "I onl=
y
use 25kV" and " an SEM will not work above 5,000X" bunch really worry me
and they should also worry their supervisors! =


Is a true scientist someone who experiments, most SEM operators do not! =

This is not a one country thing it is the same world wide, just because w=
e
are microscopists we seem to forget we are also scientists. What do the
masses think?

Steve Chapman
Senior Consultant
Protrain




From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Tue, 15 Jul 1997 15:27:00 -0500
Subject: RE: Rush Lab + Projection Slides

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} Now, with negative, flatbed scanners one could scan the negative (10
} minutes) and print an image on transparency paper with an inkjet
} (Epson 1440 dpi or a dyesub printer). Haven't tried this with TEM
} negatives but I know people who do.
}
I have no doubt that this would work, but it requires yet more capital
equipment. And here in Manitoba Canada, the money is NOT AVAILABLE.
Hence, I have to dismiss all the talk about digital images and scanners,
and that sort of thing. It's something like arguing that a Mercedes is
a nice car. I have no doubts that it is, but I nevertheless won't be
driving one in the near future, unless I win a rather large lottery.
} Seems like about one hour would do it.
}
} What about a positive TEM film that you could project directly.
}
I have not heard about the existence of this sort of TEM film, but even
if we did have the film, we would still suffer from the fact that we
would have to crop a considerable amount out of the frame in order to
get it to fit our slide projector, even with the supersize size. (ie:
2X2")

Secondly, we usually don't know ahead of time that we will need the
transparencies. The urgent images are the 8X10" prints, of which I'm
rushed to produce as fast as possible. There is no time for a
Pathologist to scope the case twice, once with positive film. And in
any event, we would need a record on negative film in any event.

I'm sure that I could develop this normal TEM film in a reversal sort of
way, in the same manner that one might work with T-MAX film developed
with reversal chemistry, but this produces positives directly, and there
is no negative produced, so that I would have the above problems, plus
the cropped images.

It seems to be quite satisfactory to produce my positive slides the way
I do, and usually speed is not a big factor with these positive images.

Garry
}
}
}
}
} ______________________________ Reply Separator
} _________________________________
} Subject: RE: Rush Lab + Projection Slides
} Author: GBurgess (GBurgess-at-exchange.hsc.mb.ca) at unix,mime
} Date: 7/14/97 2:02 PM
}
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America




From: Bennett, Cynthia, HDG / FHF :      bennett-at-msmhdg.hoechst.com
Date: Wed, 16 Jul 1997 08:40:00 +0200
Subject: AW: EDX - Spot Mode and more on SEM

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Steve,

} the biggest problem that one confronts is the "scientist" who has been =
doing
} things his way for 20 years and will not change.

That's certainly not the case for me! I'm a chemist who was been
partially using myself and partially supervising a new SEM for less than
a year. I've subscribed to the list in the hopes that I'd learn
something.

} I am interested in the size of particle that you talk about, what =
density
} do you see in your material having carried out the analysis?

} For materials similar to carbon the volume of material involved with =
an
} electron beam of an energy suitable for most EDX analysis (say 15kV) =
will
} be in excess of 2.7 microns? To carry out an analysis of 0.5 micron
} structures with material the density of carbon would require a beam =
energy
} of less than 6kV or for iron less than 10kV.

I guess I didn't express myself very clearly. We're looking at low-level
amounts of inorganic particulate contaminants or additives in plastic
(C,H,O) and what we want is QUALITATIVE information, such as "Does this
particle contain calcium or does it contain silicon?" We wouldn't dare
to try say anything about amounts (other than perhaps "lots" or "hardly
any") because we're never sure how much of the polymer matrix we're
hitting along with the particle. (The particles are at varying depths.)

In my--admittedly still amaturish--opinion, spot analysis can be OK if
what you want is qualitative information. You have to make sure that the
customers don't think the results are in any way quantitative. (They
always want to and you have to keep on banging on 'em.)

} The mark you call a "burn" is this not really contamination? There is
} usually no damage to a specimen if you see a dark mark, contamination.

I take the placement of the "burn" mark as proof that the positioning of
the beam in spot mode is OK. As to what the dark "burn" mark actually
is, I'm open. (I put those quotes on the word for a reason!) However I
think it's thermal degradation of the polymer, which melts around 260=B0 =
C
and softens and shrinks before that. I think this because we don't get
these marks when we look at metals, etc.

Cindy Bennett
Hoechst Diafoil

Disclaimer: these are my opinions only and not those of my company.






From: Garry Burgess
Date: 15 July 1997 18:12
Subject: RE: Rush Lab + Projection Slides

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Garry

we are drifting well away from the main theme, but you can still make slides
with your enlarger even if you haven't got close focussing on your lens.
It's more fiddly but if you have a light box you simply put the e.m.
negative on the light box under the enlarger and the film to be exposed in
the negative carrier.
The tricky bit is getting the position and focus right but you do most of
that by enlarging something of the right format first onto the light box.
Then of course when you're exposing the film in the enlarger you must
remember to turn on the light box and not the enlarger.

I have used this a couple of times and it works fine in an emergency
although of course if you were doing a lot it would be easier to make the
slides with a close-up 35mm camera and a light box..

Malcolm Haswell
e.m. unit
University of Sunderland
UK
----------

{SNIP}
Neat method of making slides, by the way. I'm going to remember it for
possible future use.

Yes, it works fine. But the biggest obstacle for someone doing this for the
first time is to make sure that they have a lens that is capable of making
such a small focused image. You also have to make sure that you use glass
slide holders, because negative film cannot stand up to the heat of a slide
projector without glass protection, because it's not as tough as slide film.

Garry





From: SONEJA A K :      soneja-at-giasbma.vsnl.net.in
Date: Wed, 16 Jul 1997 16:58:45 -0500 (GMT)
Subject: unsubscribe

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unsubscribe

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327 Wadala Udyog Bhavan,Wadala,MUMBAI(BOMBAY )400 031.INDIA
Tel:91 22 4145057/4165650
Fax 91 22 4168757
Email:soneja-at-giasbma.vsnl.net.in
*************************************************************************





From: Diane Montpetit :      montpetitd-at-em.agr.ca
Date: Wed, 16 Jul 1997 07:46:09 -0400
Subject: pta stain/background/formvar thank you for your suggestions

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thank you all for your suggestions,

I will tried the glow discharge and the carbon film to change the
hydrophobicity...and also putting my grids (specimen and stain) in the
oven 60C for 24 hours before looking at them to get rid of possible water
trapping.
I really appreciate your help, especially during summer time when a lot of
people are absent...

thanks again,

Diane Montpetit
microscopist
Food research center
agro alimentaire canada
st-hyacinthe, quebec, canada
tel 514 773 1105
fax 514 773 8461
e mail montpetitd-at-em.agr.ca




From: jeharper-at-amoco.com
Date: 7/15/97 2:35 PM
Subject: EDX - Spot Mode and more on SEM

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A couple of thoughts.

First, someone working 20 years in the industry is, by definition,
successful. This means that he/she is providing information that is
useful to others in solving their problems.

Second, our employers could care less about kv's, resolution,
magnification, etc. Their interest is in solving problems. Doing the
same thing for 20 years (even with a 20 year old microscope) may be
what it takes. I have seen old-timers with a 20 year old scope work
circles around newbies with newbie microscopes.

Finally, I agree whole-heartedly with Steve--we must be open to new
ideas to solve difficult problems. I worked for a major computer
manufacturer to solve a circuit board plating problem (wavy circuit
traces). They ALWAYS USED 25 kv to examine the photo resist on the
boards that produced the circuit traces. At 25 kv they looked
straight and clean. I showed them what they looked like at 5kv and at
2kv. The edges were full of unremoved resist film whose pattern
matched the wavy lines identically.

Providing INFORMATION to solve problems is our goal.



Jim Harper
Amoco Fabrics and Fibers
______________________________ Reply Separator _________________________________


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I am interested in the size of particle that you talk about, what density
do you see in your material having carried out the analysis?

For materials similar to carbon the volume of material involved with an
electron beam of an energy suitable for most EDX analysis (say 15kV) will
be in excess of 2.7 microns? To carry out an analysis of 0.5 micron
structures with material the density of carbon would require a beam energy
of less than 6kV or for iron less than 10kV.

The mark you call a "burn" is this not really contamination? There is
usually no damage to a specimen if you see a dark mark, contamination. The
problem is that with your materials you may well drive off the volatile
components and then you will burn out an area of specimen, the depth
dependant upon the time of your observation or analysis.

However you seem to have conclusive proof that you are analysing the area
that you have made your target. My point was not that a spot analysis is
taboo but that we should not blindly use a spot analysis without knowing
more about its accuracy. Some replies have taken the attitude that because
they have used the technique for many many years there cannot be a problem!
Unless you know exactly what is happening in a microscope I do not agree
that this attitude is sufficient, I can show you instruments where this
attitude would be a disaster.

In the consultancy business the biggest problem that one confronts is the
"scientist" who has been doing things his way for 20 years and will not
change. I understand that we may well compare driving a microscope to
driving a car. Who would by a new car and then go on a course to learn to
drive it? The same approach in SEM however would be constructive; new
instruments open up new techniques and new areas of investigation. From
what I see the topic of scanning electron microscopy changes considerably
within a two year cycle and operators who persist in using their 20 year
old techniques, even on an old instrument, are somewhat lost! The "I only
use 25kV" and " an SEM will not work above 5,000X" bunch really worry me
and they should also worry their supervisors!

Is a true scientist someone who experiments, most SEM operators do not!
This is not a one country thing it is the same world wide, just because we
are microscopists we seem to forget we are also scientists. What do the
masses think?

Steve Chapman
Senior Consultant
Protrain





From: Richard Beanland +44 1327 356363 :      richard.beanland-at-gecm.com
Date: Wed, 16 Jul 1997 14:23:51 +0000 (GMT)
Subject: Removing a thin TEM sample from a Cu grid

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Disclose-Recipients: prohibited

Hello all.
last week I stupidly stuck a Cu grid on top of the area of interest
of a TEM cross section. Having tried to remove it for a few days, I thought
I'd turn to the microscopy community for some help!
The sample is Si, polished to less than ten microns thick (orangey colour),
with Al and SiO2 on the top surface. It's the only one I have. I stuck the
2x1mm slot grid onto the sample with 5-minute epoxy (devcon); I didn't realise
the grid was in the wrong place until a couple of hours later. Since then
I've soaked it in dmf (dimethylformamide) [3 days] acetone [a few hours] and
ashed it in nitrogen and oxygen [a couple of hours]. The sample is still in
one piece and stuck to the grid. I've tried pushing it about with a fine hair
but it's still well fixed.
So, while it is soaking for a little longer in dmf and being ashed
periodically, has anyone got any bright ideas how to rescue my sample?

Many thanks in advance,

Richard Beanland,
Gmmt Ltd.,
Caswell,
Towcester,
Northants NN12 8EQ

Tel +44 1327 356363
Fax +44 1327 356775
e-mail richard.beanland-at-gecm.com





From: Robert H. Olley :      R.H.Olley-at-reading.ac.uk
Date: Wed, 16 Jul 1997 15:05:44 +0100 (BST)
Subject: SEM and TEM: Burns in Polymers

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To Cynthia Bennett and all Microscopy readers:

For all who are interested in burns in polymers under SEM, one does not
only get them in Spot Mode, but when looking at high magnification in
raster mode. Strange things can happen when looking at banded spherulites
of polyethylene, for example: because of the periodic variations in
crystal orientation, one gets mass transport (not mass transit, which is
the underground railway in Hong Kong!) and the banded spherulitic
structure appears to "develop" as if by etching, but what appears is in
fact an artifact which mimics the real thing.

Similar developments occur when trying to look at these beasties directly
in sections under the TEM, which is one reason why staining
(chlorosulphonic acid, RuO4) and etching (permanganic) techniques were
developed.

There is a rather poor quality picture of a banded spherulite on my home
page: URL as in the signature, but to go straight there type:

http://www.reading.ac.uk/~spsolley/pexl.html#bandsph

This was produced by etching with permanganic reagent.

+------------------------------------------------------------------------+
| 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: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Wed, 16 Jul 1997 09:13:40 -0500
Subject: RE: Permawash

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I have looked through all of my EM catalogues, checking the section on
Photography for anything like Permawash, but the only product that I can
find that comes close to that idea of hypo remover is the Kodak product
Hypo Eliminator. My problem with the Hypo Eliminator is that the
instructions suggested that the working solution only lasts 24 hours,
but I wanted to keep it in a stainless steel tank for about a week, and
just use it conveniently, without having to make up a working solution
every day.

That said, providing there were no water marks on the film, I'm still
wondering if I can rewash it later for 20 minutes, after the negatives
have already dried, and effectively remove any fixer that might have
been left in the emulsion the first time for a very abbreviated wash.

Other people have suggested that Kodak was being very conservative with
their wash times, but I'm not sure if this is true. Kodak had no
difficulty shortening the wash times for RC paper when they felt that a
2 minute wash time was sufficient for this photographic paper. So, if
they really felt that 2 or 3 minutes wash time was sufficient for their
EM film, then I would suspect that that would be their recommendation.

Garry
}
} } from what i understand, Permawash is just soapy water. i simply use a
} } mild detergent solution instead and it works fine
} }
} You may be thinking of Photoflo - which is essentially a wetting agent, a
} detergent - used to break the hydrophobicity of film and permit sheeting
} of the water. This is similar to the wetting agents used in dishwashing
} machines (e.g., for spotless glassess). Permawash should contain some
} chemical scavengers used to remove resisual fixers in the film/papers. Any
} photo-chemical types listening to this?
}
}
} ####################################################################
} John J. Bozzola, Ph.D., Director
} Center for Electron Microscopy
} Neckers Building, Room 146 - B Wing
} Southern Illinois University
} Carbondale, IL 62901-4402
} U.S.A.
} Phone: 618-453-3730
} Fax: 618-453-2665
} Email: bozzola-at-siu.edu
} Web: http://www.siu.edu/departments/shops/cem.html
} ####################################################################
}
}
}




From: Donald Lovett :      lovett-at-tcnj.edu
Date: Wed, 16 Jul 1997 11:21:49 -0400 (EDT)
Subject: Projection Slides

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The thread of this conversation appears to be drifting toward making
projection slides from EM negatives.

The technique that I have used for years is to place the EM negative (SEM
or TEM) on a light table with a cardboard mask around the negative to
block peripheral light. Use Kodak Technical Pan film in a standard
35 mm camera with a macro lens. Bracket exposures 1/2 f-stop and develop
for maximum contrast. (From start to mounted slides, less than 1 hour).

One gets crisp slides with perfect contrast (assuming that the original
negatives were good). The advantage is that one can crop the EM negative
by adjusting distance of the camera from the negative. Disadvantage is
that one cannot label the slide unless one is willing to put rub-on
letters on the EM negatives (I don't).

______________________________________________________________________
Donald L. Lovett e-mail: lovett-at-tcnj.edu
Assoc. Professor, Dept. of Biology voice: (609) 771-2876
The College of New Jersey fax: (609) 771-2674
Trenton, NJ 08650-4700







From: Dr. Mark W. Lund :      lundm-at-physc2.byu.edu
Date: Wed, 16 Jul 1997 09:53:09 MST/MDT
Subject: RE: Removing a thin TEM sample from a Cu grid

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Taking epoxy off without damaging the silicon or aluminum is tough.
Acetic acid will work but may damage the silicon film. It does not
attack bulk aluminum (much) but at the thin film level it might.
Methyl cloride is also useful for dissolving epoxy--some paint
removers will also dissolve epoxy.

best regards
mark

Mark W. Lund, PhD
Director } } Soft X-ray Web page http://www.moxtek.com { {
MOXTEK, Inc.
452 West 1260 North
Orem UT 84057 801-225-0930 FAX 801-221-1121
lundm-at-xray.byu.edu

"The state is good at simple tasks, like killing people and seizing their
wealth. It has far more trouble reaching inside individuals and making them
good." Doug Bandow




From: RWILLSON-at-pearl.tufts.edu
Date: Wed, 16 Jul 1997 11:24:36 -0500 (EST)
Subject: unsubscribe

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unsubscribe




From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Wed, 16 Jul 1997 11:27:23 -0500
Subject: Projection Slides

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Malcolm,
This sounds bizarre. Are you talking about a contact print here on to
film? Or do you mean leave the negative carrier in the carrier holder
of the enlarger, and use it upside down? (sort of like using the
enlarger as an upside down camera.)

Garry


} Garry
}
} we are drifting well away from the main theme, but you can still make slides
} with your enlarger even if you haven't got close focussing on your lens.
} It's more fiddly but if you have a light box you simply put the e.m.
} negative on the light box under the enlarger and the film to be exposed in
} the negative carrier.
} The tricky bit is getting the position and focus right but you do most of
} that by enlarging something of the right format first onto the light box.
} Then of course when you're exposing the film in the enlarger you must
} remember to turn on the light box and not the enlarger.
}
} I have used this a couple of times and it works fine in an emergency
} although of course if you were doing a lot it would be easier to make the
} slides with a close-up 35mm camera and a light box..
}
} Malcolm Haswell
} e.m. unit
} University of Sunderland
} UK
} ----------
} From: Garry Burgess
} To: 'Microscopy Society of America
} Subject: RE: Rush Lab + Projection Slides
} Date: 15 July 1997 18:12
}
} {SNIP}
} Neat method of making slides, by the way. I'm going to remember it for
} possible future use.
}
} Yes, it works fine. But the biggest obstacle for someone doing this for the
} first time is to make sure that they have a lens that is capable of making
} such a small focused image. You also have to make sure that you use glass
} slide holders, because negative film cannot stand up to the heat of a slide
} projector without glass protection, because it's not as tough as slide film.
}
} Garry
}
}




From: azita ariapour :      azita.ariapour-at-utoronto.ca
Date: Wed, 16 Jul 1997 15:48:23 -0400
Subject: unsubscribe

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From: Ron Doole :      ron.doole-at-materials.oxford.ac.uk
Date: Wed, 16 Jul 1997 18:27:37 +0100 (BST)
Subject: Re: Removing a thin TEM sample from a Cu grid

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Hi Richard,
Acetone disolves araldite so it seems a good start. Try soaking
some of your cured epoxy (not your specimen) in acetone for a while and
see if it softens, if it does then soak your specimen in it for longer. It
is probably very difficult for the acetone to get right under the copper
grid so it may need quite a long time.
Plasma ashing seems to spread epoxy around the specimen but
does not appear to remove it.

Good luck.

Ron
===========================================================================
Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk
Department of Materials, phone +44 (0) 1865 273701
University of Oxford, fax +44 (0) 1865 283333
Parks Road.
Oxford. OX1 3PH. UK.
============================================================================





From: Lesley Weston :      lesley-at-unixg.ubc.ca
Date: Wed, 16 Jul 1997 10:45:44 -0700 (PDT)
Subject: Re: Projection Slides

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To get slides with labelling, I overlay a piece of acetate with the
letters, arrows etc. rubbed onto it, placed carefully on top of the
negative. Provided the acetate is clean, it works very well. Of course, my
negatives are 3in x 4in; it might be more difficult with 35mm film.

Lesley Weston.

On Wed, 16 Jul 1997, Donald Lovett wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
}
} The thread of this conversation appears to be drifting toward making
} projection slides from EM negatives.
}
} The technique that I have used for years is to place the EM negative (SEM
} or TEM) on a light table with a cardboard mask around the negative to
} block peripheral light. Use Kodak Technical Pan film in a standard
} 35 mm camera with a macro lens. Bracket exposures 1/2 f-stop and develop
} for maximum contrast. (From start to mounted slides, less than 1 hour).
}
} One gets crisp slides with perfect contrast (assuming that the original
} negatives were good). The advantage is that one can crop the EM negative
} by adjusting distance of the camera from the negative. Disadvantage is
} that one cannot label the slide unless one is willing to put rub-on
} letters on the EM negatives (I don't).
}
} ______________________________________________________________________
} Donald L. Lovett e-mail: lovett-at-tcnj.edu
} Assoc. Professor, Dept. of Biology voice: (609) 771-2876
} The College of New Jersey fax: (609) 771-2674
} Trenton, NJ 08650-4700
}
}
}
}





From: Damian_Neuberger_at_RLT013-at-ccmailgw.mcgawpark.baxter.com
Date: Wed, 16 Jul 1997 12:23:05 -0500
Subject: Storing in fixative.

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Hi everyone,

Quick question:

1. How long can I store rat tissue in glutaraldehyde fixative before
conpleting the tissue preparation process and not experience tissue
degradation? Can I go as long as three weeks? At this point, I don't
know if I will be processing for SEM (CPD) or TEM - depends on the LM
results.

2. Should I store in buffer rather than the fixative? Some other
solution?

3. Would I use a different formulation of fixative for this kind of
delayed processing of tissue?

So much for quick questions and thanks so much for your help! Now, I
have to go answer someone else's question.

Damian Neuberger
neuberd-at-baxter.com




From: Damian_Neuberger_at_RLT013-at-ccmailgw.mcgawpark.baxter.com
Date: Wed, 16 Jul 1997 12:23:05 -0500
Subject: Storing in fixative.

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

Hi everyone,

Quick question:

1. How long can I store rat tissue in glutaraldehyde fixative before
conpleting the tissue preparation process and not experience tissue
degradation? Can I go as long as three weeks? At this point, I don't
know if I will be processing for SEM (CPD) or TEM - depends on the LM
results.

2. Should I store in buffer rather than the fixative? Some other
solution?

3. Would I use a different formulation of fixative for this kind of
delayed processing of tissue?

So much for quick questions and thanks so much for your help! Now, I
have to go answer someone else's question.

Damian Neuberger
neuberd-at-baxter.com




From: South Bay Technology :      73531.1344-at-CompuServe.COM
Date: 16 Jul 97 13:59:07 EDT
Subject: Re: Removing a thin TEM sample from a Cu grid

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Micro Listserver {microscopy-at-Sparc5.Microscopy.Com}

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Dear Richard:

I was intrigued by your question and decided to contact Devcon directly. I was
told by their applications engineer that the 5 minute epoxy will withstand DMF,
acetone and most anything else. He said the only thing it doesn't stand up too
well against is water!

He suggested soaking the sample in warm water for a few hours and then prying it
apart with a razor blade. When I explained how fragile the sample was, he just
said to soak it longer and it will come apart. If you still have trouble
getting it apart, try contacting Devcon directly. They have a help section on
their web site - unfortunately, I forgot to copy down their web site address,
but I found it just by searching on "Devcon".

Please let me know if the water works.

Best regards-

David

+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++

David Henriks TEL: 800-728-2233 (toll-free in USA)
South Bay Technology, Inc. 714-492-2600
1120 Via Callejon FAX: 714-492-1499
San Clemente, CA 92673 USA e-mail: 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 Richard Beanland +44 1327 356363
}
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Hello all.
last week I stupidly stuck a Cu grid on top of the area of interest
of a TEM cross section. Having tried to remove it for a few days, I thought
I'd turn to the microscopy community for some help!
The sample is Si, polished to less than ten microns thick (orangey colour),
with Al and SiO2 on the top surface. It's the only one I have. I stuck the
2x1mm slot grid onto the sample with 5-minute epoxy (devcon); I didn't realise
the grid was in the wrong place until a couple of hours later. Since then
I've soaked it in dmf (dimethylformamide) [3 days] acetone [a few hours] and
ashed it in nitrogen and oxygen [a couple of hours]. The sample is still in
one piece and stuck to the grid. I've tried pushing it about with a fine hair
but it's still well fixed.
So, while it is soaking for a little longer in dmf and being ashed
periodically, has anyone got any bright ideas how to rescue my sample?

Many thanks in advance,

Richard Beanland,
Gmmt Ltd.,
Caswell,
Towcester,
Northants NN12 8EQ

Tel +44 1327 356363
Fax +44 1327 356775
e-mail richard.beanland-at-gecm.com {





From: James Martin :      James.S.Martin-at-williams.edu
Date: Wed, 16 Jul 1997 14:25:58 -0400 (EDT)
Subject: forensic/materials science, video-digital imaging

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I am planning to purchase a video or digital camera to document,
analyze, and print images of paint and fiber samples using polarized light
microscopy and fluorescence microscopy. Previous threads re video or
digital cameras have focused largely on black and white images obtained
using SEM or TEM and printed using high-DPI inkjets or dye-sub printers.

May I ask for recommendations or comments on purchasing a system (input
through output) to deal with colored images of paint cross-sections,
fibers, petrographic samples, etc.? This system would allow still images
to be captured, analyzed/manipulated using image analysis software
(including Photoshop), embedded in reports, and printed with near
photographic quality in color.

I'll be pleased to provide more details, as requested.

TIA.

James Martin





From: Bernard Kestel :      bernard_kestel-at-qmgate.anl.gov
Date: 16 Jul 1997 13:29:49 -0500
Subject: JET ELECTROPOLISHING

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Continuing the discussion of this topic. I used twin jet electropolishing
a bit about 1970, but was frustrated by not being able to determine when a
shiny surface was produced. (I had been able to see the sample in an old glass
system operated manually). About that time, one of the early South Bay 550
polishers was ordered by me here at Argonne because it permitted magnified, IN
SITU, viewing of the sample during polishing. This was needed to thin 1 or 2
new materials every week! The ease of use permitted accumulating
reproducible data published in a 65 page report, (ANL-80-120), used world
wide, a journal cover photo, and 12 other published articles. This work
brought me the MSA "Technologist of the Year" award in 1994. About six 550
polishers are used exclusively at Argonne-some as long as 25 years!
The unit can do jetting from one side, (for back-thinning to a special
surface-lacquer protected), or both sides by inverting the sample after jet
polishing about half way thru it. Microshield lacquer, (from South Bay),
works great to protect surfaces from etching, dissolves in acetone, and may be
thinned to reduce shrinkage when thinning soft, annealled copper for example.
Also, the entire 3 m.m. disc surface can be polished via a 3 m.m. jet; using
an external "timer/switch",and a D.C. power supply, planar "sectioning" of as
little as 100 nm. can be removed from a surface. The jet polishing
electrolyte and conditions will work. The large jet can be used to etch a
surface for optical photos by simply reducing the "polishing" voltage about
20% for a couple of seconds! The 300 volt, 150 mA. capacity power supply
exeeds other manufacturer's units and makes use of non-acid "BK-2" type
electrolytes possible-a must for many materials. The line-of-sight optical
shut off system can be fitted with a variety of color spectrum light sources
for special uses. The standard infra red LED and detector bias may be
independently adjusted to give the desired sensitivity setting. It will make
electron transparant regions in pure annealled metal such as aluminum-with no
hole! Of course the setting is normally set for a 20 micron hole with a very
thin edge (quite reproducible, of course). Alignment of the parts is easy and
stays set a long time. Even saphire light pipes are available for
hydrofluoric acid or bromine/alcohol solutions. PVC plastic parts are
available and may be substituted for metal ones for such strong chemical
baths. Low temperatures of -50 degrees C. are no problem. The sample is
accesible for rapid rinsig after swinging the detent-equipped jet support to
one side.
In 25 years of use, these instruments have saved one man per year in labor
cost, (roughly $100,000/yr.), or $2,500,000--due to the ease and speed with
which excellent samples can be made. About 90 to 95% of the samples attempted
are good once- conditions are established.
In my opinion, all the jet polishers have improved with time, but the South
Bay 550 C and 550 D units are unmatched when it comes to working with the
newer, difficult materials which should be viewd DURING thinning. They permit
me to thin about 300 TEM foils/year in my spare time.





From: rtind-at-siu.edu (Randy Tindall)
Date: Wed, 16 Jul 1997 13:51:51 -0600
Subject: Projection Slides

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Might as well throw in my two cents worth on this topic. We
generally make our projections slides from prints, rather than negatives.
This, of course, requires making prints first, but since this is often done
anyway, it's usually not a problem.

We use a seldom-mentioned film known as Kodak 5468, a direct
positive film used, I think, as a motion picture stock. It's bright red
and translucent---strange looking stuff. Our exposures on a 4-light copy
stand range from 6-10 seconds at a lens opening of f/3.5 on a Canon 55mm
macro lens, so it obviously requires a lot of light. Development is in
Dektol diluted 1:1 with water. Yes, Dektol, the paper developer. Stop
with water, fix with whatever you usually use and wash normally.

This film is cheap (still less than $25/100 ft., last I checked),
development is easy, and the slides are quite good. The problem is that
this film tends to undergo reduction-oxidation (redox) after a few years,
giving a solarized appearance. To prevent this, use Kodak Brown Toner
(TOXIC---use plenty of ventilation and gloves) diluted about 1:50 for a
30-second dip. This will often give your slides a slight brown tone. Some
folks like this, some don't.

Not a perfect solution, but very useful for many purposes.

Randy Tindall
Center for Electron Microscopy
Southern Illinois University at Carbondale






From: Winston Wiggins :      wwiggins-at-mail.carolinas.org
Date: 7/16/97
Subject: RE: Permawash

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Gerry et al.,
Perma Wash is advertised as an archival high speed wash for both film
and paper that reduces wash time by 90%. It's made by Heico (a Cambrex Co.)
along with other kinds of photographic chemicals.
I've used it for years both as an EM specialist and as a photographer
with no problems at all. You should check with local photographic darkroom
supply house instead of any EM suppliers. Two suppliers that I know who do
offer it are listed if you still can't find it in your area.
For film Heico states archival permanence with one minute first
water wash, one minute Perma Wash, one minute final water wash; for r.c. paper,
it's two minutes, two minutes, two minutes; finally, for double weight papers,
five minutes, five minutes, five minutes does the job.
I hope this washes well for you. :-) :-) :-}

Camera World Crimson Tech
PO Box 9426 325 Vassar Street
1809 Commonwealth Ave Cambridge, MA 02139
Charlotte, NC 28205
Pho: 800-868-3686 800-868-5150
704-375-8453 617-868-5150
Fax: 704-376-1826 617-499=4777

------------------------------------
Name: Winston Wiggins
E-mail: wwiggins-at-carolinas.org





From: tom moninger :      moninger-at-emiris.iaf.uiowa.edu
Date: Wed, 16 Jul 1997 14:07:30 -0500 (CDT)
Subject: RE: Permawash

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FYI,

PERMA WASH Archival High Speed Wash

HEICO Chemicals Inc.
Route 611
Delaware Water Gap, PA 18327

717-420-3900

Contains ammonium sulfite, sodium sulfite and water.

The container say it specifically removes silver sulfate.

Tom

Thomas Moninger moninger-at-emiris.iaf.uiowa.edu
University of Iowa Central Microscopy Research Facility
http://www.uiowa.edu/~cemrf
Views expressed are mine.





From: William Tivol :      tivol-at-wadsworth.org
Date: Wed, 16 Jul 1997 17:31:24 -0500 (EDT)
Subject: Re: Removing a thin TEM sample from a Cu grid

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} last week I stupidly stuck a Cu grid on top of the area of interest
} of a TEM cross section. Having tried to remove it for a few days, I thought
} I'd turn to the microscopy community for some help!
} The sample is Si, polished to less than ten microns thick (orangey colour),
} with Al and SiO2 on the top surface. It's the only one I have. I stuck the
} 2x1mm slot grid onto the sample with 5-minute epoxy (devcon); I didn't real-
} ise the grid was in the wrong place until a couple of hours later. Since
} then I've soaked it in dmf (dimethylformamide) [3 days] acetone [a few
} hours] and ashed it in nitrogen and oxygen [a couple of hours]. The sam-
} ple is still in one piece and stuck to the grid. I've tried pushing it
} about with a fine hair but it's still well fixed.
} So, while it is soaking for a little longer in dmf and being ashed
} periodically, has anyone got any bright ideas how to rescue my sample?
}
} Many thanks in advance,
}
} Richard Beanland,


} I was intrigued by your question and decided to contact Devcon directly.
} I was told by their applications engineer that the 5 minute epoxy will with-
} stand DMF, acetone and most anything else. He said the only thing it does-
} n't stand up too well against is water!
} He suggested soaking the sample in warm water for a few hours and then pry-
} ing it apart with a razor blade. When I explained how fragile the sample
} was, he just said to soak it longer and it will come apart.

} David
}
Dear Richard,
Here's a thought: If just soaking won't do, you might try micro-
waving the water-soaked specimen. That will heat up the water, but maybe
not the Si, Al or Cu. I know metal is a no-no in a microwave oven, but
there is so little that there shouldn't be a disaster here. I'd put ~50
ml of H2O in a beaker in the oven at the same time. Of course, I'd try
it first with something other than the specimen. Good luck.
Yours,
Bill Tivol







From: Bernard Kestel :      bernard_kestel-at-qmgate.anl.gov
Date: 16 Jul 1997 16:46:01 -0500
Subject: JET ELECTROPOLISHING

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Continuing the discussion of this topic. I used twin jet electropolishing
a bit about 1970, but was frustrated by not being able to determine when a
shiny surface was produced. (I had been able to see the sample in an old glass
system operated manually). About that time, one of the early South Bay 550
polishers was ordered by me here at Argonne because it permitted magnified, IN
SITU, viewing of the sample during polishing. This was needed to thin 1 or 2
new materials every week! The ease of use permitted accumulating
reproducible data published in a 65 page report, (ANL-80-120), used world
wide, a journal cover photo, and 12 other published articles. This work
brought me the MSA "Technologist of the Year" award in 1994. About six 550
polishers are used exclusively at Argonne-some as long as 25 years!
The unit can do jetting from one side, (for back-thinning to a special
surface-lacquer protected), or both sides by inverting the sample after jet
polishing about half way thru it. Microshield lacquer, (from South Bay),
works great to protect surfaces from etching, dissolves in acetone, and may be
thinned to reduce shrinkage when thinning soft, annealled copper for example.
Also, the entire 3 m.m. disc surface can be polished via a 3 m.m. jet; using
an external "timer/switch",and a D.C. power supply, planar "sectioning" of as
little as 100 nm. can be removed from a surface. The jet polishing
electrolyte and conditions will work. The large jet can be used to etch a
surface for optical photos by simply reducing the "polishing" voltage about
20% for a couple of seconds! The 300 volt, 150 mA. capacity power supply
exeeds other manufacturer's units and makes use of non-acid "BK-2" type
electrolytes possible-a must for many materials. The line-of-sight optical
shut off system can be fitted with a variety of color spectrum light sources
for special uses. The standard infra red LED and detector bias may be
independently adjusted to give the desired sensitivity setting. It will make
electron transparant regions in pure annealled metal such as aluminum-with no
hole! Of course the setting is normally set for a 20 micron hole with a very
thin edge (quite reproducible, of course). Alignment of the parts is easy and
stays set a long time. Even saphire light pipes are available for
hydrofluoric acid or bromine/alcohol solutions. PVC plastic parts are
available and may be substituted for metal ones for such strong chemical
baths. Low temperatures of -50 degrees C. are no problem. The sample is
accesible for rapid rinsig after swinging the detent-equipped jet support to
one side.
In 25 years of use, these instruments have saved one man per year in labor
cost, (roughly $100,000/yr.), or $2,500,000--due to the ease and speed with
which excellent samples can be made. About 90 to 95% of the samples attempted
are good once- conditions are established.
In my opinion, all the jet polishers have improved with time, but the South
Bay 550 C and 550 D units are unmatched when it comes to working with the
newer, difficult materials which should be viewd DURING thinning. They permit
me to thin about 300 TEM foils/year in my spare time.





From: Tom Phillips :      tphillips-at-biosci.mbp.missouri.edu
Date: Wed, 16 Jul 1997 17:20:48 -0500
Subject: Au labeling of membranes - can you judge sideness?

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We are having a mini-debate here over whether one can confidently say what
side of the membrane an epitope on a membrane faces (e.g., towards the
cytoplasm or lumen of the RER) based on the distribution of gold labeling.
If most the labeling is on one side, would you be confident the epitope is
solely on that side? One of my collaborators had a paper criticized by a
reviewer who said this couldn't be done reliably.


Thomas E. Phillips, Ph.D.
Associate Professor of Biological Sciences
Director, Molecular Cytology Core Facility
3 Tucker Hall
University of Missouri
Columbia, MO 65211
(573)-882-4712 (voice)
(573)-882-0123 (fax)






From: Dr. Mark W. Lund :      lundm-at-physc2.byu.edu
Date: Wed, 16 Jul 1997 16:33:16 MST/MDT
Subject: RE: Removing a thin TEM sample from a Cu grid

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OOPs--I intended to say that acetic acid may damage the
aluminum film. It doesn't attack silicon from my experience.

best regards
mark

Mark W. Lund, PhD
Director } } Soft X-ray Web page http://www.moxtek.com { {
MOXTEK, Inc.
452 West 1260 North
Orem UT 84057 801-225-0930 FAX 801-221-1121
lundm-at-xray.byu.edu

"The state is good at simple tasks, like killing people and seizing their
wealth. It has far more trouble reaching inside individuals and making them
good." Doug Bandow




From: David Griffiths :      DGriffiths-at-SAS.Samsung.com
Date: Wed, 16 Jul 1997 18:18:22 -0500
Subject: Position Available -- Entry Level Auger Technician

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} The Company:
}
} Samsung Austin wants to offer you more than a job. We want to offer you the
} chance to develop a career. Becoming involved in a fast-paced start-up
} operation is both exciting and challenging and we look for employees who are
} energetic, flexible, and team-oriented.
} Are you willing to go the extra mile? Are you comfortable with change? Are
} you interested in great benefits? Are you excited about working in a start-up
} environment? Are you interested in a career where you will be an integral
} part of a new company? Are you comfortable making decisions that will
} influence the development of our corporate culture?
} If the answer to these questions is "yes," then Samsung Austin is the company
} for you.
} For more information visit our web site at www.sas.samsung.com
}
}
} The Position
}
} We are currently looking to hire an Entry Level Auger Technician. The
} requirements for this position is simply an Associate Degree in a technical
} major. Experience is preferred but not necessary. The position will involve
} shift work.
}
} If someone know of a person who would consider this position or if you know
} of a 2 year college that offers a course which covers Auger Electron
} Spectroscopy please feel free to contact me.
}
} Resumes can be faxed or emailed to:
} David A. Griffiths
} Fax (512)491-1165
} Phone/VoiceMail(512)491-1403
} Pager (512)209-4132
} e-mail DGriffiths-at-sas.samsung.com






From: Garber, Charles A. :      cgarber-at-2spi.com
Date: Wed, 16 Jul 97 21:26:44 -0500
Subject: Bringing a TEM sample back to life

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

Richard Beanland described a problem involving a TEM grid stick to a sample
in the wrong place, glued with a 5-Minute (Devcon) epoxy.

One is his comments was:
================================================
I've soaked it in dmf (dimethylformamide) [3 days] acetone [a few hours] and
ashed it in nitrogen and oxygen [a couple of hours]. The sample is still in
one piece and stuck to the grid. I've tried pushing it about with a fine
hair but it's still well fixed.
================================================
Oxygen plasma etching, in an isotropic (I stress isotropic as opposed to
anisotropic) plasma etcher should remove the epoxy. If you were correct in
that you used "nitrogen and oxygen", then this is why it did not work. You
have to use pure oxygen, no nitrogen. Even a small leak in the system,
allowing just a small partial pressure of nitrogen will literally kill the
etching rate. In air, literally nothing will happen.

Now, if in fact you were using pure oxygen, there could still be a leak
problem, another reason why you did not get any etching. But the technique
should work. Make sure the power is not more than 100 watts or else the
sample might heat up to temperature that would not be acceptable.

Disclaimer: SPI manufactures an isotropic plasma etcher for doing this kind
of etching of organic materials. You can see further information on our
website as well as an explanation of the differences between isotropic vs.
anisotripic etching.

Chuck

==================================================
Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400
President 1-(800)-2424-SPI
SPI SUPPLIES FAX: 1-(610)-436-5755
PO BOX 656 e-mail: cgarber-at-2spi.com
West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com


Look for us!
############################
WWW: http://www.2spi.com
############################
==================================================




From: Craig Lending :      clending-at-acs.brockport.edu
Date: Wed, 16 Jul 1997 21:28:00 -0400
Subject: Tetrahymena fixation protocol -- Help, please

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Does anyone have a tried and true method for the fixation and embedment
of Tetrahymena?
I would like to do some preembedding staining for nuclear proteins, and
have been having trouble once the dehydration is started. The cells are
blasted apart by the time I examine them under the electron microscope.
The cells are intact after preembedding immunostaining (at least they
appear to be under phase contrast). After a post immunostaining
treatment with 1% glutaraldehyde, the cells are enrobed in agar (1:1 of
2% low-melting temperature agar) at 37C, dehydrated in a graded ethanol
series (10, 30, 50, 75, 95, 100, 100 -- 15 minutes each) and then
progressively infiltrated with LR White resin (all at room temp).
Polymerization is at 55C for 24 hours.

Any help/ideas would be greatly appreciated.

Craig Lending
Department of Biology
SUNY Brockport
Brockport, NY 14420
Phone: 716-395-5755
Fax: 716-395-2741
e-mail: clending-at-acs.brockport.edu





From: Jim Darley :      jim-at-proscitech.com.au
Date: Thu, 17 Jul 1997 09:02:27 +1000
Subject: re: storing in fixative

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Damian -
Sabatini (he made GA the EM fixative) did some experiments and
declared that postfixation could be left for at least six months. I guess
that is true for a few tissues. Lipids are not well fixed in GA and lipid
rich tissues in particular suffer when post fixation is delayed.
By how much - well how long is a piece of string? Postfix as soon as
possible. Store in buffer refrigerated.
What you do for SEM or LM matters very little, but for TEM this matters.
Never store tissues in GA for TEM. GA crosslinks materials, overfixing with
GA results in a coarser texture which obscures fine details at high
resolution.
Jim Darley

ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313
Great microscopy catalogue, 400+ Links, MSDS
************************ http://www.proscitech.com.au
}
} 1. How long can I store rat tissue in glutaraldehyde fixative
before
} conpleting the tissue preparation process and not experience tissue
} degradation? Can I go as long as three weeks? At this point, I
don't
} know if I will be processing for SEM (CPD) or TEM - depends on the
LM
} results.
}
} 2. Should I store in buffer rather than the fixative? Some other
} solution?
}
} 3. Would I use a different formulation of fixative for this kind of

} delayed processing of tissue?
}
} So much for quick questions and thanks so much for your help! Now,
I
} have to go answer someone else's question.
}
} Damian Neuberger
} neuberd-at-baxter.com





From: Mary Mager :      mager-at-unixg.ubc.ca
Date: Wed, 16 Jul 1997 20:31:04 -0700
Subject: Re: Removing a thin TEM sample from a Cu grid

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Dear Richard,
The only way I have removed 5-minute epoxy from something was by gently
heating. The epoxy softens at heat-gun temps (70 deg.C?). This was on a
leaky air-pressure valve, not a grid, so I just heated it until I could peel
it off.. The other suggestion would be acetone, at least overnight. If you
ask a embedding/biologist EM type, they may know a solvent to dissolve epoxy.
You wrote:

} Hello all.
} last week I stupidly stuck a Cu grid on top of the area of interest
} of a TEM cross section. Having tried to remove it for a few days, I thought
} I'd turn to the microscopy community for some help!
} The sample is Si, polished to less than ten microns thick (orangey colour),
} with Al and SiO2 on the top surface. It's the only one I have. I stuck the
} 2x1mm slot grid onto the sample with 5-minute epoxy (devcon); I didn't realise
} the grid was in the wrong place until a couple of hours later. Since then
} I've soaked it in dmf (dimethylformamide) [3 days] acetone [a few hours] and
} ashed it in nitrogen and oxygen [a couple of hours]. The sample is still in
} one piece and stuck to the grid. I've tried pushing it about with a fine hair
} but it's still well fixed.
} So, while it is soaking for a little longer in dmf and being ashed
} periodically, has anyone got any bright ideas how to rescue my sample?
}
} Many thanks in advance,
}
} Richard Beanland,

Regards,
Mary






From: wesleysm-at-biology.und.ac.za (James Wesley-Smith)
Date: Thu, 17 Jul 1997 08:08:38 +-200
Subject: LM coverslip mounting medium

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Good day everyone
I would appreciate your comments on what permanent mounting media are
available which meet the correct refractive index of glass and do not lead
to fading of toluidene blue-stained sections?

Thank you.


James Wesley-Smith
EM Unit
University of Natal
Durban, South Africa





From: ROBIN CROSS :      EURC-at-giraffe.ru.ac.za
Date: Thu, 17 Jul 1997 08:34:36 GMT+0200
Subject: Re: Projection Slides

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Kodak Rapid Process Copy film (if it is still available) or
Kodak Direct MP film (is available, e.g. from SPI or Ted
Pella, etc) are excellent for inexpensive single-process
preparation of B & W transparencies from EM prints.


Robin H Cross
Director : EM Unit, Rhodes University, Grahamstown, South Africa
eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377




From: Seppo J. Sivonen :      ssivonen-at-sun3.oulu.fi
Date: Thu, 17 Jul 1997 12:27:20 +0300
Subject: EDX - Spot Mode - Small Particles

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Here is one addition (hopefully valuable) to the EDX-Spot Mode discussion.
=20
If your SEM has a specimen current meter, you can use that successfully
for checking the beam position very accurately before and during the EDX
analysis of small particles.=20

The technique is very simple. You first position the spot mode beam
at high magnification, and then by moving the beam in X-Y direction
you either maximize or minimize the specimen current reading depending on
whether the average atomic number (=3D Z) of the particle is lighter or=20
heavier that that of the matrix. The great advantage in the use of the=20
specimen current (=3D absorbed electrons) for positioning the beam is that=
=20
you same time maximize the X-ray emission from the particle and minimize
the possible X-ray contribution from the matrix. This is due to the fact
that absorbed electrons "sense" the shape of the particle under the specimen
surface.

If the specimen current stays constant during the measurement of the EDX
spectrum, then you can be absolutely certain that the beam did not leave
the particle during the measurement. However, normally there is a small
increase ( { 1 %) in the specimen current due to the contamination build-up.






=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=
=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=
=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4
=A4 =A4
=A4 Seppo J. Sivonen e-mail: seppo.sivonen-at-oulu.fi =A4=20
=A4 University of Oulu =A4=20
=A4 Institute of Electron Optics tel: +358-8-553 3140 =A4
=A4 Box 400 fax: +358-8-553 3149 =A4=20
=A4 FIN-90571 Oulu =A4
=A4 FINLAND http://koivu.oulu.fi/~eolwww/welcome.html =A4
=A4 =A4
=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=
=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=
=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4




From: Stephen Griffiths :      s.griffiths-at-ucl.ac.uk
Date: Thu, 17 Jul 1997 10:37:32 +0100
Subject: Re: LM coverslip mounting medium

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Hi

I've had this problem of fading sections too.

I don't know about the refractive index, but I've used Epoxy resin as a
mountant quite successfully. It doesn't seem to fade Toluidine Blue
semi-thins.

DePeX is not too bad but I have had some fading over long periods.

Regards
Stephen

{} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}
Stephen Griffiths e-mail:- s.griffiths-at-ucl.ac.uk
Visual Science Department Phone:- 0171 608 6914
Institute of Ophthalmology Fax:- 0171 608 6850
Bath Street, London. EC1V 9EL
{} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}

----------
} From: James Wesley-Smith {wesleysm-at-biology.und.ac.za}
} To: 'Microscopy-at-sparc5.microscopy.com' {Microscopy-at-Sparc5.Microscopy.Com}
} Subject: LM coverslip mounting medium
} Date: 17 July 1997 09:08
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Good day everyone
} I would appreciate your comments on what permanent mounting media are
} available which meet the correct refractive index of glass and do not
lead
} to fading of toluidene blue-stained sections?
}
} Thank you.
}
}
} James Wesley-Smith
} EM Unit
} University of Natal
} Durban, South Africa
}




From: Lesley Suzanne Bechtold :      lsb-at-aretha.jax.org
Date: Thu, 17 Jul 1997 08:08:11 -0400
Subject: Decalcification

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Hi,

I'm looking for some information or references on decalcifying with
ascorbic acid versus EDTA for TEM samples.

Thanks in advance!

Lesley Bechtold





From: Owen P. Mills :      opmills-at-mtu.edu
Date: Thu, 17 Jul 1997 08:07:43 -0400
Subject: Polaron sputter target

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Hello everyone,

Can anyone recommend a vendor to supply a Cu target for a Polaron E5000
sputter coater?
Thanks.

Owen


Owen P. Mills
Michigan Technological University
Metallurgical & Materials Engineering
Rm 512 MME Building
Houghton, MI 49931
906-487-2002
906-487-2934 FAX
opmills-at-mtu.edu




From: Garry Burgess
Date: 16 July 1997 17:42
Subject: Projection Slides

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Perhaps I didn't make my original comments clear enough. But yes the
unexposed negative goes in the enlarger film carrier in the enlarger. This
has the big advantage that reduction is no problem. The disadvantage is that
if your enlarger doesn't do one to one printing in the first place then you
still can't do it - just the reductions. This is usually not a problem if
you are doing this with e.m cut film because it is larger and so needs to be
reduced. You could even do this with prints ie copying them from the
baseboard of the enlarger onto the negative in the film carrier but the
amounts of stray light needed for copying stand lights makes it more fiddly.
I think someone else has already mentioned it is important to mask out stray
light.

I stumbled upon this idea because in one lab, I worked, they had a large
Durst Laborator 1000 enlarger with all the accessories. This allowed you to
use it as a copy camera and gave you special large format film holders for
the purpose. Another lab I worked in had a DeVere large format enlarger with
full reduction facilities. It seemed reasonable therefore to use an ordinary
Durst enlarger as a copy camera so that I could do the 'DeVere thing, in
reverse. I just didn't have all of the clever light-tight film holders for
masking unexposed film in the carrier but with care it works, anyway.

If I remember rightly many of the big Durst enlargers have a reversible
mirror in the condensor system - our Durst 1200s look as if you can just
take out the fitting so that it faces towards the operator rather than the
lamp. You should then be able to view the image of your negative or whatever
else is on the baseboard through the little window in the front of your
enlarger and VOILA you have a simple camera which will do large format. I am
sure it should be in the manual somewhere.

Bizarre it may seem, to use an enlarger to reduce, but isn't there a certain
pleasing symmetry to it?
Sorry to go on but I thought this might be of general interest.

DISCLAIMERS:
I hasten to add you can only do this with some enlargers; it will disrupt
normal printing in a one enlarger darkroom; you may damage the enlarger so
be careful; if in doubt read the manual or ask the supplier/manufacturer;
and I will refuse to re-imburse anyone for self-inflicted damage.
I have no connections with Durst or De Vere other than as a satisfied user.

Malcolm Haswell
----------


Malcolm,
This sounds bizarre. Are you talking about a contact print here on to
film? Or do you mean leave the negative carrier in the carrier holder
of the enlarger, and use it upside down? (sort of like using the
enlarger as an upside down camera.)

Garry


} Garry
}
} we are drifting well away from the main theme, but you can still make
slides
} with your enlarger even if you haven't got close focussing on your lens.
} It's more fiddly but if you have a light box you simply put the e.m.
} negative on the light box under the enlarger and the film to be exposed in
} the negative carrier.
} The tricky bit is getting the position and focus right but you do most of
} that by enlarging something of the right format first onto the light box.
} Then of course when you're exposing the film in the enlarger you must
} remember to turn on the light box and not the enlarger.
}
} I have used this a couple of times and it works fine in an emergency
} although of course if you were doing a lot it would be easier to make the
} slides with a close-up 35mm camera and a light box..
}
} Malcolm Haswell
} e.m. unit
} University of Sunderland
} UK
} ----------
} From: Garry Burgess
} To: 'Microscopy Society of America
} Subject: RE: Rush Lab + Projection Slides
} Date: 15 July 1997 18:12
}
} {SNIP}
} Neat method of making slides, by the way. I'm going to remember it for
} possible future use.
}
} Yes, it works fine. But the biggest obstacle for someone doing this for
the
} first time is to make sure that they have a lens that is capable of making
} such a small focused image. You also have to make sure that you use glass
} slide holders, because negative film cannot stand up to the heat of a slide

} projector without glass protection, because it's not as tough as slide
film.
}
} Garry





From: Simon C. Watkins :      swatkins-at-pitt.edu
Date: Thu, 17 Jul 1997 08:11:51 -0400
Subject: Re: Au labeling of membranes - can you judge sideness?

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Message-ID: {33CE0C07.AFCB262F-at-pitt.edu}

Tom I tend to agree with the reviewer. I have spent several years
looking at the membrane cytroskeleton of muscle, and commonly the gold
particles appear "extracellular" when we know for sure they are on the
inside of the plasma membrane. The argument is that as an IgG is about
11nm long, tagged to a 5nm gold particle, and if an indirct tagging
system is system is used this adds an another 11nm. this leads of a
potential radius of 25nm from the label site. I am sure from
quantitative studies it is less than this, though I am equally sure that
the gold particle commonly does not reflect to epitope position exactly.

Simon

Tom Phillips wrote:

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} We are having a mini-debate here over whether one can confidently say
} what
} side of the membrane an epitope on a membrane faces (e.g., towards the
}
} cytoplasm or lumen of the RER) based on the distribution of gold
} labeling.
} If most the labeling is on one side, would you be confident the
} epitope is
} solely on that side? One of my collaborators had a paper criticized
} by a
} reviewer who said this couldn't be done reliably.
}
} Thomas E. Phillips, Ph.D.
} Associate Professor of Biological Sciences
} Director, Molecular Cytology Core Facility
} 3 Tucker Hall
} University of Missouri
} Columbia, MO 65211
} (573)-882-4712 (voice)
} (573)-882-0123 (fax)



--
Simon C. Watkins Ph.D.
Associate Professor
Director CBI
University of Pittsburgh
Pittsburgh PA 15261
tel:412-648-3051
Fax:412-648-2004
URL:http://sbic6.sbic.pitt.edu






From: Linda Iadarola :      Linda.Iadarola-at-quickmail.yale.edu
Date: 17 Jul 1997 08:45:45 -0400
Subject: Re: Au labeling of membranes

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"Tom Phillips" {tphillips-at-biosci.mbp.missouri.edu}
X-Mailer: Mail*Link SMTP-QM 3.0.3 b1 d5



From: RWILLSON-at-pearl.tufts.edu
Date: Thu, 17 Jul 1997 08:58:06 -0500 (EST)
Subject: unsubscribe

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Reply to: RE} Au labeling of membranes - can you judge sideness?

Dear Thomas,
Your discussion regarding which side of the membrane the labeling is on should
include how to determine the resolution of the labeling technique. The
resolution is determined by the size of the probe used for visualization as well
as the antibody used. For example, an IgG molecule has an extended length of
about 8-10nm and a 5nm gold particle would be expected to have a total diameter
of 7-8nm of covered with protein-A. The resolution comes from the circular
radius of this complex from the antigen outward and upwards. The antibody could
fall anywhere within this radius, thus making it difficult to say if it is on
the inside or outside of the membrane. You could confidently say it labels the
membrane but to go further may be stretching what you see visually. Section
thickness also plays a part in reducing the resolution. For further information
you may want to pick up a book by Garreth Griffiths called "Fine structure
immunocytochemistry" printed by Springer-Verlag ISBN 3-540-54805-X. It is a
comprehensive look at a number of variables pertaining to immunocytochemistry.

Linda Chicoine
Center for Cell Imaging
Dept. of Cell Biology
Yale University
203-785-3646 phone
203-785-7226 fax

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

We are having a mini-debate here over whether one can confidently say what
side of the membrane an epitope on a membrane faces (e.g., towards the
cytoplasm or lumen of the RER) based on the distribution of gold labeling.
If most the labeling is on one side, would you be confident the epitope is
solely on that side? One of my collaborators had a paper criticized by a
reviewer who said this couldn't be done reliably.


Thomas E. Phillips, Ph.D.
Associate Professor of Biological Sciences
Director, Molecular Cytology Core Facility
3 Tucker Hall
University of Missouri
Columbia, MO 65211
(573)-882-4712 (voice)
(573)-882-0123 (fax)



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Could someone please tell me how to unsubscribe to the Microscopy
List Server? I plan to be a way for the next couple of weeks and would like
to turn the flood of messages off.
Thanks
Rob Willson
Dept of Anatomy and Cell Biology
Tufts University




From: Louisa.Howard-at-Dartmouth.EDU (Louisa Howard)
Date: 17 Jul 97 09:14:00 EDT
Subject: Re: Au labeling of membranes - can you judge sideness?

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Tom Phillips wrote:
....."what side of the membrane an epitope on a membrane faces (e.g., towards
thecytoplasm or lumen of the RER) based on the distribution of gold
labeling."....

I would also agree that the normal gold label may be to large, because of the
size of the coupled IgG moelcule. What about the Nanogold labels: 1.4 nm gold
attched to FAB fragment? I don't know what the total size would be, but much
smalelr than if coupled to igG.Would this be small enough? the silver
enhancement, to enlarge the size for viewing on TEM, is done after label.
Louisa Howard
EM Facility, 6044 Gilman
Dartmouth College
Hanover, NH, 03755




From: C.John Runions :      cjr14-at-cornell.edu
Date: Thu, 17 Jul 1997 09:49:18 +0500
Subject: Re: LM coverslip mounting medium

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=================
C. John Runions, Ph.D
Section of Ecology and Systematics
Corson Hall
Cornell University
Ithaca, New York
USA 14853

email cjr14-at-cornell.edu [ie. cjr(fourteen)-at-...]
phone (607) 254-4282
Fax (607) 255-8088






From: marshall-at-uimrl7.mrl.uiuc.edu (mike marshall)
Date: Thu, 17 Jul 1997 08:50:08 -0500
Subject: Re: Removing a thin TEM sample from a Cu grid

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Well, i dont know if this is a dumb solution, but you could try to etch the
copper away with somehting like dilute nitric acid. i dont know the
dangers with silicon or the aluminum, but perhaps you could find an etchant
that would attack only the copper.

Michael T. Marshall
Research Engineer, Electron Microscopy
University of Illinois at Urbana-Champaign
Frederick Seitz Materials Research Laboratory
104 South Goodwin avenue
Urbana, IL 61801-2985
(217) 244-8193 fax: (217) 244-2278






From: Damian_Neuberger_at_RLT013-at-ccmailgw.mcgawpark.baxter.com
Date: Thu, 17 Jul 1997 08:45:38 -0500
Subject: Permawash

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For what it's worth regarding the use of Permawash: I have negatives
that I washed according to their (Permawash) instructions at least 25
years ago and they are still in perfectly good condition.

Damian Neuberger
neuberd-at-baxter.com

Usual disclaimer....




From: Damian_Neuberger_at_RLT013-at-ccmailgw.mcgawpark.baxter.com
Date: Thu, 17 Jul 1997 09:23:47 -0500
Subject: Summary-store in fix.

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Thanks to all who replied to my question about storing rat tissue in
fixative. Here is a cut and paste summary of replies from all over
the world! for anyone who is interested.

Sabatini (he made GA the EM fixative) did some experiments and
declared that postfixation could be left for at least six months. I
guess that is true for a few tissues. Lipids are not well fixed in GA
and lipid rich tissues in particular suffer when post fixation is
delayed. By how much - well how long is a piece of string? Postfix as
soon as possible. Store in buffer refrigerated.
What you do for SEM or LM matters very little, but for TEM this
matters. Never store tissues in GA for TEM. GA crosslinks materials,
overfixing with GA results in a coarser texture which obscures fine
details at high resolution.

I have stored samples in 2% glut. in 0.1M sodium cacodylate for 2
weeks because I forgot about them. These were cell cultures and they
turned out OK, I embedded these into epon/araldite. I have also
stored things in buffer after fixation for a few weeks and this turned
out OK too.

Dysktra's book on biological EM has micrographs of mouse kidney stored
in formaldehyde/glut fix for something like 2 years that looks pretty
good.


I often have kept things in GA for as long as 3 weeks, provided
that it is kept cold. (There may be some problem with microtubule
disassembly, according to some). Over time, the GA will break down
3weeks).

Store in buffer: No. You stand the risk of fixative being washed out
and the possibility of "de-fixing" the material. You also could
encourage bacterial growth eventually.

My recommendation would be to fix, wash, and post-fix in OsO4. If
you then dehydrate to 70% EtOH or Acetone,it will keep for YEARS!

Although we try not to store anything, we prefer to store our samples
in Trumps fix. A buffered glute/form combo. We have stored for weeks
at a time and had good results

There is some evidence to suggest that storage in Trumps or half
Karnovsky rather than pure glut (i.e. glut plut some paraformaldehyde)

I have stored tissue in glutaraldehyde or Trump's fix(glut/para fix)
for years before using it for TEM. I wouldn't store it in buffer.

I would probably do a normal fix and then switch to buffer and store
cold. I don't believe you will see any degradation as long as you stay
away from post-fixation with osmium before storage; I would leave
that go until you were ready to proceed with prep.

Damian Neuberger
neuberd-at-baxter.com




From: MIKE ROCK :      merock-at-du.edu
Date: Thu, 17 Jul 1997 09:19:57 -0600 (MDT)
Subject: Re: Storing in fixative.

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Damian-
I've always tried to avoid any long term storage in fix, however storing
samples after fixation in a 0.2M buffer has seemed to work well for up to
1-2 weeks. And after reading the thread on lipid preservation, I think
it wise to osmicate prior to the storage in buffer too.
_mike




From: Caroline Schooley :      schooley-at-mcn.org
Date: Thu, 17 Jul 1997 08:37:51 -0700
Subject: Re: LM coverslip mounting medium

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} } Good day everyone
} } I would appreciate your comments on what permanent mounting media are
} } available which meet the correct refractive index of glass and do not
} lead
} } to fading of toluidene blue-stained sections?
} }
I learned a trick from a DuPont-Sorvall rep (that tells you how long ago
that was!) that retards or eliminates ALL fading caused by oxidants in the
mounting medium. Add 1-2% BHT (the preservative used in bologna, hot dogs,
etc.) to any mounting medium. There should be a bottle of the stuff
sitting on the shelf in the EML at U.C. Berkeley that has enough to supply
every EM lab in the country...

Caroline Schooley
Educational Outreach Coordinator
Microscopy Society of America
Box 117, 45301 Caspar Point Road
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html
Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/






From: Caroline Schooley :      schooley-at-mcn.org
Date: Thu, 17 Jul 1997 08:37:51 -0700
Subject: Re: LM coverslip mounting medium

Contents Retrieved from Microscopy Listserver Archives
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} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

} } Good day everyone
} } I would appreciate your comments on what permanent mounting media are
} } available which meet the correct refractive index of glass and do not
} lead
} } to fading of toluidene blue-stained sections?
} }
I learned a trick from a DuPont-Sorvall rep (that tells you how long ago
that was!) that retards or eliminates ALL fading caused by oxidants in the
mounting medium. Add 1-2% BHT (the preservative used in bologna, hot dogs,
etc.) to any mounting medium. There should be a bottle of the stuff
sitting on the shelf in the EML at U.C. Berkeley that has enough to supply
every EM lab in the country...

Caroline Schooley
Educational Outreach Coordinator
Microscopy Society of America
Box 117, 45301 Caspar Point Road
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html
Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/






From: Gary Liechty :      garyliechty-at-worldnet.att.net
Date: Thu, 17 Jul 1997 08:58:01 -0700
Subject: Re: Removing a thin TEM sample from a Cu grid

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Richard Beanland +44 1327 356363 wrote:
}
} ------------------------------------------------------------------------
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} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Hello all.
} last week I stupidly stuck a Cu grid on top of the area of interest
} of a TEM cross section. Having tried to remove it for a few days, I thought
} I'd turn to the microscopy community for some help!
} The sample is Si, polished to less than ten microns thick (orangey colour),
} with Al and SiO2 on the top surface. It's the only one I have. I stuck the
} 2x1mm slot grid onto the sample with 5-minute epoxy (devcon); I didn't realise
} the grid was in the wrong place until a couple of hours later. Since then
} I've soaked it in dmf (dimethylformamide) [3 days] acetone [a few hours] and
} ashed it in nitrogen and oxygen [a couple of hours]. The sample is still in
} one piece and stuck to the grid. I've tried pushing it about with a fine hair
} but it's still well fixed.
} So, while it is soaking for a little longer in dmf and being ashed
} periodically, has anyone got any bright ideas how to rescue my sample?
}
} Many thanks in advance,
}
} Richard Beanland,
} Gmmt Ltd.,
} Caswell,
} Towcester,
} Northants NN12 8EQ
}
} Tel +44 1327 356363
} Fax +44 1327 356775
} e-mail richard.beanland-at-gecm.com
Dear Mr. Beanland,

We have available an Epoxy Dissolver that is supposed to work and all 2
component epoxies. As I have not tried this product on all epoxies
available, I do not know if it will work with this particular type.

If you can, visit your local pharmacy and ask them for some
DMSO(Dimethylsulfoxide). This is the primary ingredient and it will
need to be heated to operate effectively.

Please let me know if you have any other questions.

Sincerely,

Gary Liechty
Allied High Tech Products, Inc.
2376 E. Pacifica Pl.
Rancho Dominguez, Ca. 90220
310-635-2466
800-675-1118
310-762-6808 Fax

Products for Materialographic, SEM and TEM sample preparation




From: Bernard Kestel :      bernard_kestel-at-qmgate.anl.gov
Date: 17 Jul 1997 11:24:53 -0500
Subject: P.S. , SINGLE JET

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From: Bernie Kestel-Argonne National Lab

I forgot to mention an important feature on the South Bay 550 series jet
polishers in my previous message. The "open faced" specimen retainer has only
a 0.0015" thick polyethylene diaphram with a central hole in it to hold the
specimen down on a platinum tipped holder. This thin sheet offers almost NO
flow resistance or bubble trapping area near the specimen. The all important
electrolyte viscosity/polishing film can be thicker with this design,
producing smoother finished surfaces while "bridging" across grain boundaries,
precipitates and other features. That is why I do most polishing at -45 C. or
so and add butyl cellosolve to increase electrolyte viscosity to 10-12
centipoises-ideal. (Like half & half from a refrigerator, approx.). Of course
a PVC cap with a central hole positions the specimen laterally.
I pass this along to hopefully ease someones prep. burden - -I'm NOT
financially connected to South Bay! I feel this unit is like driving a modern
auto compared to a hand cranked, manual shifted, no air conditioning machine.
Take the easy route!





From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Thu, 17 Jul 1997 11:40:30 -0500
Subject: Expired Glutaraldehyde

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Message-Id: {c=CA%a=_%p=Health_Sciences_%l=POSTOFFICE-970717164030Z-4622-at-postoffice.hsc.mb.ca}

Our lab has been using Glutaraldehyde that expired in November of 1995,
but has been kept refrigerated since then. Lately we have been noticing
problems with our fixation, and I was wondering if perhaps it is because
of this old Glutaraldehyde. Does anyone know how important these expiry
dates really are with respect to its effect on fixation?????????

Not properly fixed,

Garry




From: Michael L. Mead :      michaelmead-at-sprintmail.com
Date: Thu, 17 Jul 1997 14:13:54 -0700
Subject: Re: unsubscribe

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To All:

I don't mean to be hypercritical but doesn't everybody else see how to
unsubscribe at the top of their message. I am a new subscriber (one
week) and there must have been at least 3 messages similar to this one.
Read the rules or does the old adage apply that:

OLD MICROSCOPISTS NEVER DIE
THEY JUST LOSE THEIR RESOLUTION.

Mike Mead







RWILLSON-at-pearl.tufts.edu wrote:

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} America
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} Could someone please tell me how to unsubscribe to the Microscopy
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} to turn the flood of messages off.
} Thanks
} Rob Willson
} Dept of Anatomy and Cell Biology
} Tufts University



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To All:

{P} I don't mean to be hypercritical but doesn't everybody else see how
to unsubscribe at the top of their message.  I am a new subscriber
(one week) and there must have been at least 3 messages similar to this
one.  Read the rules or does the old adage apply that:

{P} {FONT SIZE=+2} OLD MICROSCOPISTS NEVER DIE {/FONT}
{BR} {FONT SIZE=+2} THEY JUST LOSE THEIR RESOLUTION. {/FONT} {FONT SIZE=+2} {/FONT}

{P} {FONT SIZE=+2} Mike Mead {/FONT}
{BR}  
{BR}  
{BR}  
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{P} RWILLSON-at-pearl.tufts.edu wrote:
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{P}  Could someone please tell me how to unsubscribe to the Microscopy
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{BR}  Rob Willson
{BR} Dept of Anatomy and Cell Biology
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From: Geoff McAuliffe :      mcauliff-at-UMDNJ.EDU
Date: Thu, 17 Jul 1997 15:59:28 -0700
Subject: Re: more on storage in fixative

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Dear All:

After reading the summary Damian posted I wanted to chip in my 2
cents worth.

One respondent recommeded storage in fix since long-term buffer storage
might "unfix" the specimen. I find it hard to believe that buffer could
undo the powerful crosslinking glut causes. I have heard of this
possibility with formalin fixation but that is not nearly as powerful a
fix as glut. Anyone know of any experiments in this area?

Another responsent advocated storage in ethanol or acetone after
post-fixing in OsO4, stating the tissue would keep for years. It will
keep but a lot of cytoplasm will be extracted. Hayat's book, Prin. and
Tech. of EM (1981 edition) discusses this and gives references and an
illustration on pp 150-155.

I would store in buffer, changing it often if I could not complete
processing soon.

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 e-mail: mcauliff-at-umdnj.edu
***************************************************************




From: EUGENE GORDON :      MEDJET-at-worldnet.att.net
Date: Thu, 17 Jul 1997 17:15:50 -0500
Subject: Ultracentrifuge and oven

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Hello all,

I am in the market for a used clinical ultracentrifuge as well as an oven
to be used during TEM specimen embedding. If anyone knows of a lab that is
interested in selling these items, please contact me directly.

Thanks,

Dan Caruso c/o Eugene Gordon
Biological Technician
Medjet, Inc.
1090 King Georges Post Road
Edison, NJ 08837
Phone: (732) 738-3990
Fax: (732) 738-3984
MEDJET-at-WORLDNET.ATT.NET






From: Garber, Charles A. :      cgarber-at-2spi.com
Date: Thu, 17 Jul 97 19:01:01 -0500
Subject: Expired Glutaraldehyde

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

Garry Burgess wrote:
==================================================
Our lab has been using Glutaraldehyde that expired in November of 1995, but
has been kept refrigerated since then. Lately we have been noticing
problems with our fixation, and I was wondering if perhaps it is because of
this old Glutaraldehyde. Does anyone know how important these expiry dates
really are with respect to its effect on fixation??????
==================================================
You are quite correct in that there can be some degree of arbitrariness in
the statement of the expiration date. At least to us, the expiration date
should correlate with some future point in time, after which, one could
expect to see some deterioration of performance. Of course many of us know
that film and paper, if properly stored don't turn into pumpkins on their
expiration dates.

In the case of glutaraldehyde, the two most important factors influencing
what will be the actual expiration date (as opposed to that stamped on the
product), in the case of glut would be

a) starting purity of the ampouled product, since it is an autocatalytic
reaction, and once the dimers and trimers reach some critical level,
deterioration (e.g. polymerization) can proceed quite quickly. Hence a
starting purity of the least amounts of the dimers and trimers, etc.
relative to a glut with higher levels, would be expected to have longer
shelf life. But we are talking about variations in starting purities that,
when fresh, I would expect, would give any user good results.

b) thermal history during shipment. It is quite an education to follow the
progress of a shipment and to see to what levels of heat exposure a
particular shipment is exposed. Over the years I have myself "visited" UPS
and FedEx trucks and have been quite surprised at how hot the inside of a
truck can get on a hot summer day. Indeed some of the large warehouse type
sorting rooms of the courier services are not air conditioned. Travelling
on a highway for hours at a time with a hot summer sun beating down on the
trailer leads to sometimes very hot temperatures. I have also been in
institutional receiving departments that have been like a furnace in the
middle of the summer. And I have been in receiving departments during the
winter, and on the coldest of days, when supplemental heat is being provided
by portable space heaters and the temperature of nearby boxes seem almost
too hot to touch (well a slight exaggeration, because they were not breaking
out into flames, but you get my point).

You would not want to know what a receiving department is like at Cairo
University in June and July, where the temperatures are over 100 deg. in the
shade!

So the answer is, you don't know what has been that thermal history. Have
you been lucky or have you been unlucky? Most people would not want to
leave the outcome of their important experiments to the chance that it was
OK.

So at least in our case, and I suspect others too, supplier applied
expiration dates are a best estimate to predict when one should start
thinking about ordering fresh material and tossing the old material.

Chuck

PS: In the specific case at hand, if there is even the slightest trace of a
precipitate in the ampoule, just write it off and don't even try using it.
But in this case, even if there was not a precipitate, that fact we are
approaching two years beyond the expiration date (e.g. 11/95), it would be
good cause the do the same thing.

===================================================
Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400
President 1-(800)-2424-SPI
SPI SUPPLIES FAX: 1-(610)-436-5755
PO BOX 656 e-mail: cgarber-at-2spi.com
West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com


Look for us!
############################
WWW: http://www.2spi.com
############################
==================================================




From: Andrew Buechele :      andrew-at-rsrch.vsl.cua.edu
Date: Thu, 17 Jul 1997 19:39:38 EST
Subject: Re: Removing a thin TEM sample from a Cu grid

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I don't have any experience with using it on Devcon 5 minute epoxy,
but I have found that DMSO will dissove several epoxies quite well.
Observe the usual precautions about DMSO, i.e. keep it off your
hands, especially when it may contain any toxic substance.

Sincerely,
Andy
Andy Buechele
The Catholic University of America
409 Hannan Hall
Washington, D.C. 20064
(202) 319-4995 FAX: (202) 319-4469




From: Andrew Buechele :      andrew-at-rsrch.vsl.cua.edu
Date: Thu, 17 Jul 1997 20:42:12 EST
Subject: Re: Removing a thin TEM sample from a Cu grid

Contents Retrieved from Microscopy Listserver Archives
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I don't have any experience with using it on Devcon 5 minute epoxy,
but I have found that DMSO will dissove several epoxies quite well.
Observe the usual precautions about DMSO, i.e. keep it off your
hands, especially when it may contain any toxic substance.

Sincerely,
Andy


Andy Buechele
The Catholic University of America
409 Hannan Hall
Washington, D.C. 20064
(202) 319-4995 FAX: (202) 319-4469




From: Jim Darley :      jim-at-proscitech.com.au
Date: Fri, 18 Jul 1997 14:47:36 +1000
Subject: Re: storing in fixative

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George & www:
These things were published over twenty years ago. What concerns me about
our microscopy forum is that many of us are re-iterating believes or
quoting from memory.
Looking things up is considerable work and books do not include all
important knowledge previously published.
Here is a bit of my memory: I recall a discussion between Sjostrand and
Cosslett almost 30 years ago. Sjostrand had published biological sections
and claimed 10 A resolution and told the group that he was going to reduce
that substantially. Cosslett then got up and with a few figures proved (?)
that 10 A was as well as could be done with fixed tissues in sections.
Later there were publications showing that in monolayers, cells required
only three minutes of 1% (?) GA fixation. Beyond that, over fixation caused
artefact - meaningless granularity, which obscured the finest details.
Nothing to affect the average x30k micrograph, but at 300k its a big
factor.
Over fixing is not a good practise.
Jim Darley

ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313
Great microscopy catalogue, 400+ Links, MSDS
************************ http://www.proscitech.com.au


----------
} From: George C. Ruben {George.C.Ruben-at-Dartmouth.EDU}
} To: jim-at-proscitech.com.au
} Subject: re: storing in fixative
} Date: Thursday, 17 July 1997 21:39
}
} --- You wrote:
} Never store tissues in GA for TEM. GA crosslinks materials, overfixing
with
} GA results in a coarser texture which obscures fine details at high
} resolution.
} --- end of quote ---
}
} What embedding resolution do we we have if fixation is done correctly
and does
} overfixing effect resolution? Have you run standards or are you just
talking
} about the qualitative details in a picture after post fixing and
staining--
}
} --George C. Ruben
} Dept . Biological Sciences




From: chiba-at-mimj.co.jp (Chiba Atsushi)
Date: Fri, 18 Jul 1997 15:37:57 +0900
Subject: Pupil projection lens?

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Could anyone tell me what to call a lens that is located between a galvano-scanner and objective lens in the light path of a scanning laser microscope?

I'm translating a manual of a scanning laser microscope from Japanese into English. (Is it *scanning laser* microscope or *laser scanning* microscope anyway?) And I can't find a proper English term for this lens which is called "pupil projection lens" whe
n translated literally.

Any suggestion is welcomed.

Thanks in advance.


Chiba Atsushi [(Mr.) -- *Chiba* is my surname]
Voice: (+81) 010-045-9451






From: Milos Motejl :      motejl-at-paru.cas.cz
Date: Fri, 18 Jul 1997 08:44:34 +0200 (MET DST)
Subject: Low-dose for Philips420

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Dear Microscopists,
we plann to use TEM Philips 420 for Low-dose work. We will need Low-dose
system for this microscope. Unfortunately Philips said us,that they didn't
produce Low-dose for this kind of microscope,already.
Can you help us please, how or where we could get this Low-dose unit or
how to work in low-dose conditions /to prevent destroing samples during
focusing/ without this system on TEM Philips 420 ?
All your responces or opinions will wery useful for us.

Thank You very much
Milos

Milos Motejl
Lab. of Biomembranes
South Bohemian University
Ceske Budejovice
Czech Republic

motejl-at-paru.cas.cz
tel. 042-038/7775485
................................





From: awilson-at-sghms.ac.uk (A.Wilson)
Date: Fri, 18 Jul 97 10:27:32 BST
Subject: decalcification for EM

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Dear Lesley and other interested people,

I tested EDTA versus Chromium Potossium Sulphate and found the latter much
better for ultrastructural preservation (I was looking at
bone-lining-cells) as long as the pieces of bone were tiny and the decalc
short (few days) I have the original reference somewhere around if a
medline search doesn't do the trick. Dunno about ascorbic acid as I
didn't try that.

Amanda

Miss A.J.Wilson
Electron Microscope Unit
St George's Hospital Medical School
Cranmer Terrace
Tooting London SW17 ORE
Tel: 0181 725 5220
awilson-at-sghms.ac.uk
awilson-at-aw.u-net.com






From: Tony Bruton :      bruton-at-EMU.UNP.AC.ZA
Date: Fri, 18 Jul 1997 11:25:39 +0200
Subject: LKB knifebreaker, service -Reply

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Linda

We have two LKB 7800 series Knifemakers that still render outstanding
service. Please have a look at our publication "further modification of the
LKB 7800 series Knifemaker for improved reproducibility in breaking'cryo'
knives.(ref Jnl of Microscopy Vol. 168, Pt 1. Nov 1992 pp 111 - 114). The
simple modifications suggested in this paper transformed our
knifemaker's such that any thought of replacing them with newer 'better'
units vaporised. I remember that Jan Slot, on a visit to our lab, was very
impressed with the modification and performance of the knifemakers
some years back

Tony Bruton
University of Natal
Pietermaritzburg
South Africa

} } } Linda Fox {lfox1-at-wpo.it.luc.edu} 2/July/1997 06:57pm } } }
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Does anyone know where we can get our LKB 7800B knife breaker
serviced? If possible, by someone in the Chicago area? It is making
sporadically bad knives and we have adjusted all the knobs by the
instruction booklet.
If it needs to be replaced, can anyone recommend a good one? This
one has been with us for over 20 years.

Thanks, Linda Fox, Loyola Univ. Medical Center, Chicago
lfox1-at-wpo.it.luc.edu





From: wesleysm-at-biology.und.ac.za (James Wesley-Smith)
Date: Fri, 18 Jul 1997 12:04:27 +-200
Subject: Freeze frac. attachment Edwards 306

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Hi again

Has anyone out there had first hand experience using the freeze fracture
attachment for an Edwards 306 vacuum coating unit? Please reply directly to
me.

Thank you very much.

James Wesley-Smith
Electron Microscope Unit
University of Natal
Durban, South Africa






From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Fri, 18 Jul 1997 09:34:41 -0500
Subject: Expired Glut. - Another Question

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First of all, I'd like to thank all of those who cared to respond to my
question, especially Dr. Garber from SPI who gave a detailed and
thoughtful response. And OK, I get the message, it's just not worth the
risk.

But I have another question that perhaps you people also might have some
thoughts on. If I am understanding them properly, some of the
Pathologists here claim that they can tell the difference between a
specimen that was delayed before putting into Glut., resulting in
artifact such as swollen mitochondria, vs. poor fixation as a result of
outdated glutaraldehyde, such as damaged membranes in general. Is this
sort of reasoning valid?

Just curious,
Garry




From: kna101-at-utdallas.edu
Date: Fri, 18 Jul 1997 09:43:31 -0500 (CDT)
Subject: Re: Expired Glutaraldehyde

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To all,

I agree with Dr. Garber, there is a good possibility the fix has
gone off. If I were you, I would order a small replacement supply and
test this to make sure your problem is the fix- that way you assure
yourself of not wasting your supply. Order more and throw out the old
glut. once your certain it's the problem.

My two cents,

Karen Pawlowski
Lab. Tech.
UT Southwestern Medical Center,
PhD Student
UT Dallas, Dallas TX




From: WARRENJ1-at-cliffy.polaroid.com
Date: Fri, 18 Jul 1997 08:52 -0400 (EDT)
Subject: Re: forensic/materials science, video-digital imaging

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======================================================================
==========

James:

Are you by chance the Jim Martin I know with MA State Police?

I have been working this year to help bring the new Polaroid "DMC"
Digital Microscope Camera to market. If we can confirm your location,
I would recommend arranging a demo of this camera which should be
EXCELLENT for all but the very lowest-level fluorescence work. Also,
we will be featuring the DMC at the IAI meeting at Danvers, MA if
you're going.

You'll need a WIN-95/Pentium system (Mac version drivers on the way in
about 30-days). Recommend at least 32mb RAM and a hard drive large
enough to handle your library of images (1.3mb or 5.5mb uncompressed,
depending on resolution selected). The system plugs-in directly to
any software that is TWAIN compatible (that's almost anything!).

Printers? Anything you want. Dye-sub is generally the best quality,
but I have had outstanding success on the ink jet printers (like HP
870, et. al.) if the best media are used. Try the "premium glossy"
papers, or even the "premium" clay-coated (matte finish) papers.

Get back to me with questions: corlb-at-polaroid.com
Also there are specifications on the Polaroid Web Page:
www.polaroid.com, look under "Polaroid at work".



______________________________ Reply Separator
_________________________________ Subject: forensic/materials science,
video-digital imaging
Author: John D Warren at ~575ts2
Date: 7/16/97 6:35 PM


----------------------------------------------------------------------
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-.

I am planning to purchase a video or digital camera to document,
analyze, and print images of paint and fiber samples using polarized
light microscopy and fluorescence microscopy. Previous threads re
video or digital cameras have focused largely on black and white
images obtained using SEM or TEM and printed using high-DPI inkjets or
dye-sub printers.

May I ask for recommendations or comments on purchasing a system
(input through output) to deal with colored images of paint
cross-sections, fibers, petrographic samples, etc.? This system would
allow still images to be captured, analyzed/manipulated using image
analysis software (including Photoshop), embedded in reports, and
printed with near photographic quality in color.

I'll be pleased to provide more details, as requested.

TIA.

James Martin




From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Fri, 18 Jul 1997 12:07:06 -0500
Subject: Expired Glut. - 1 More thing

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First of all, I'd like to thank all of those who cared to respond to my
question, especially Dr. Garber from SPI who gave a detailed and
thoughtful response. And OK, I get the message, it's just not worth the
risk.

But I have another question that perhaps you people also might have some
thoughts on. If I am understanding them properly, some of the
Pathologists here claim that they can tell the difference between a
specimen that was delayed before putting into Glut., resulting in
artifact such as swollen mitochondria, vs. poor fixation as a result of
outdated glutaraldehyde, such as damaged membranes in general. Is this
sort of reasoning valid?

Just curious,
Garry






From: Jane M. Woodruff :      polysci-at-tigger.jvnc.net
Date: Fri, 18 Jul 1997 14:22:31 -0400
Subject: Re: Expired Glutaraldehyde

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We have two grades of glutaraldehyde - EM & BIO, and I believe you
are talking about the former.

We have done some study on storage and the polymer peak at
230nm, and found material in a sealed ampule staying alright for
up to two years. But since the customer will be breaking open
an ampule, take out to lab, withdraw a little, and then put it back
in the refrigerator, we figured these "in's and out's" will cut the
stability to some extent and put an expiry date of one year.

Assuming that your lot with an expiration of November, 1995, was
not taken out and then put back into the refrig too frequently,
it could be stable for at least six more months; i.e.,
June, 1996, if not until the end of 1996. But now it is
well beyond that period and so has possibly some polymer
which causes it to be less effective in fixation. You should
probably consider buying a new lot.

Good luck!

DR. PARASARAN
POLYSCIENCES, INC.

----------
} From: Garry Burgess {GBurgess-at-exchange.hsc.mb.ca}
} To: 'Microscopy Society of America - Mailing List'
{microscopy-at-sparc5.microscopy.com}
} Subject: Expired Glutaraldehyde
} Date: Thursday, July 17, 1997 12:40 PM
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Our lab has been using Glutaraldehyde that expired in November of 1995,
} but has been kept refrigerated since then. Lately we have been noticing
} problems with our fixation, and I was wondering if perhaps it is because
} of this old Glutaraldehyde. Does anyone know how important these expiry
} dates really are with respect to its effect on fixation?????????
}
} Not properly fixed,
}
} Garry




From: Warren Straszheim :      wes-at-ameslab.gov
Date: Fri, 18 Jul 1997 11:49:57 -0500
Subject: Re: EDX - Spot Mode - Small Particles

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Along the same lines, we use the brightness of the video signal to make sure
we are on the feature of interest. Our JEOL 840A has LED meters for contrast
and brightness that we can monitor even in spot mode. We even used to do
this on our JEOL U3, but it has been gone so long now I cannot remember
exactly how we watched for it.

At 12:27 PM 7/17/97 +0300, you wrote:
} Here is one addition (hopefully valuable) to the EDX-Spot Mode discussion.
}
} If your SEM has a specimen current meter, you can use that successfully
} for checking the beam position very accurately before and during the EDX
} analysis of small particles.
}
} The technique is very simple. You first position the spot mode beam
} at high magnification, and then by moving the beam in X-Y direction
} you either maximize or minimize the specimen current reading depending on
} whether the average atomic number (= Z) of the particle is lighter or
} heavier that that of the matrix. The great advantage in the use of the
} specimen current (= absorbed electrons) for positioning the beam is that
} you same time maximize the X-ray emission from the particle and minimize
} the possible X-ray contribution from the matrix. This is due to the fact
} that absorbed electrons "sense" the shape of the particle under the specimen
} surface.
}
} If the specimen current stays constant during the measurement of the EDX
} spectrum, then you can be absolutely certain that the beam did not leave
} the particle during the measurement. However, normally there is a small
} increase ( { 1 %) in the specimen current due to the contamination build-up.
----------------------------------------------------
Warren E. Straszheim
270 Metals Development, Ames Lab/ISU, Ames IA, 50011
Phone: 515-294-8187 FAX: 515-294-3091

E-Mail: wes-at-ameslab.gov (or: wesaia-at-iastate.edu)
http://www.public.iastate.edu/~iprt_info/cfce/ (re: coal)
http://www.public.iastate.edu/~wesaia/marl/ (re: SEM)

electron microscopy, x-ray analysis, image analysis
computer applications
coal characterization and processing





From: Dr. Mark W. Lund :      lundm-at-physc2.byu.edu
Date: Fri, 18 Jul 1997 14:49:25 MST/MDT
Subject: RE: Pupil projection lens?

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Nora,

We examine similar samples in our lab on occasion. I would need more
information on your samples to give a more specific procedure, but
here are a few tips:

a) pigments in mineral oil: try diluting sample about 1:20 in hexane,
heptane, or mineral spirits (i.e., a compatible solvent), let pigments
settle out by gravity overnight (if they will), otherwise spin down
gently in a centrifuge. Decant off the supernatant (i.e., the mineral
oil in the solvent) without losing the pigments. Add more solvent to
the container and try to redisperse the pigments. With some method
development, you should be able to obtain a dispersion of the pigment
in the solvent with only a little of the mineral oil left. Then put a
droplet of this dispersion on a suitable polished substrate (carbon?),
and wick off a little of the solvent with a kimwipe.

b) pigments in an oil/water emulsion. If you mean that the sample is
an oil in water emulsion like a latex, dilute the sample in water
1:20, put a droplet on the substrate, and wick off the water. In this
way you should be able to separate the 'oil' and the pigments enough
to pick out the pigments and image or analyze them.

Dave Klimovich
Sherwin-Williams Co.

Disclaimer: All opinions, etc., are my own.


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,
Dear friends from the list,


We have just received an enquiry about SEM analysis
for: a) pigments particles dispersed in mineral oils
b) pigments particles in oil-water emulsions

Does any of you have any experience in that
subject? Unfortunately, we do not count with equipment for sample preparation
other than the sputter-coater and carbon evaporator.

Any help will be very welcome.

Thanks in advance,

Nora Pratta
Centro Regional de Investigacion y Desarrollo
Santa Fe - Argentina





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Chiba Atsushi wrote:
:Could anyone tell me what to call a lens that is located between a galvano-scanner and objective lens in the light path of a scanning laser microscope?
:
:I'm translating a manual of a scanning laser microscope from Japanese into English. (Is it *scanning laser* microscope or *laser scanning* microscope anyway?) And I can't find a proper English term for this lens which is called "pupil projection lens" wh
e
:
:Any suggestion is welcomed.
:
:Thanks in advance.

There are several words that can be used, but "transfer lens" is very common.
Also used are "telecentric lens" and "relay lens."

best regards
mark

Mark W. Lund, PhD
Director } } Soft X-ray Web page http://www.moxtek.com { {
MOXTEK, Inc.
452 West 1260 North
Orem UT 84057 801-225-0930 FAX 801-221-1121
lundm-at-xray.byu.edu

"The state is good at simple tasks, like killing people and seizing their
wealth. It has far more trouble reaching inside individuals and making them
good." Doug Bandow






From: Michael L. Mead :      michaelmead-at-sprintmail.com
Date: Fri, 18 Jul 1997 17:40:36 -0700
Subject: Expired Glut.& Swollen Mitochondria

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In response to Garry Burgess's question regarding swollen mitochondria
sometimes referred to as "popcorn" mitochondria, the only time I have
seen them perfectly preserved was after whole body perfusion of fixative
through the heart.

It is likely that pathologists do not have tissue preserved in this way
from diagnosing human diseases. I guess for argument sake, Garry, if I
were in your shoes I would ask the pathologist (tactfully) weather the
swollen mitochondria could be the result of the pathology and not poor
fixation.

For perspective, though, I should tell you that I once worked in the
Anatomy Department at Loma Linda University and the professor there
didn't believe in purified glutaraldehye. He used the 25% stuff and
kept it under the lab bench. His belief was that the older it got the
better....something to do with the ratio of dimer to trimer. If you
want to check his publications, I believe his name was Robert Schultz.

Michael Mead

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{HTML}
{FONT SIZE=+1} In response to Garry Burgess's question regarding swollen
mitochondria sometimes referred to as "popcorn" mitochondria, the only
time I have seen them perfectly preserved was after whole body perfusion
of fixative through the heart. {/FONT} {FONT SIZE=+1} {/FONT}

{P} {FONT SIZE=+1} It is likely that pathologists do not have tissue preserved
in this way from diagnosing human diseases.  I guess for argument
sake, Garry, if I were in your shoes I would ask the pathologist (tactfully)
weather the swollen mitochondria could be the result of the pathology and
not poor fixation. {/FONT} {FONT SIZE=+1} {/FONT}

{P} {FONT SIZE=+1} For perspective, though, I should tell you that I once
worked in the Anatomy Department at Loma Linda University and the professor
there didn't believe in purified glutaraldehye.  He used the 25% stuff
and kept it under the lab bench.  His belief was that the older it
got the better....something to do with the ratio of dimer to trimer. 
If you want to check his publications, I believe his name was Robert Schultz. {/FONT} {FONT SIZE=+1} {/FONT}

{P} {FONT SIZE=+1} Michael Mead {/FONT} {/HTML}

--------------15FA38525522D4AC6C4A80FB--





From: Michael L. Mead :      michaelmead-at-sprintmail.com
Date: Fri, 18 Jul 1997 21:43:13 -0700
Subject: Re: Expired Glut. Another Question

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--------------CEF267DADC33931662890F41
Content-Type: text/plain; charset=us-ascii
Content-Transfer-Encoding: 7bit

-----------------------------------------------------------------------
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-----------------------------------------------------------------------.

In response to Garry Burgess's question regarding swollen mitochondria
sometimes referred to as "popcorn" mitochondria, the only time I have
seen
them perfectly preserved was after whole body perfusion of fixative
through
a living and pumping heart.

Obviously, it is not likely that pathologists have tissue preserved in
this way
from living humans. I guess for argument sake, I would ask the
pathologist
weather the swollen mitochondria and other membrane defects could be the

result of the pathology and not poor fixation.

On the other hand, dead cells such as cuticle and cortical cells in
hair have
well defined cell membrane complexes that look well preserved even if
you
don't fix the hair--although cell organelles like mitochondria are
missing.

For perspective, I should tell you that I once worked in the Anatomy
Department at Loma Linda University and the professor there didn't
believe
in purified glutaraldehye. He used the 25% stuff and kept it under the
lab bench.
His belief was that the older it got the better....something to do with
the ratio of
dimer to trimer. I'm not a chemist, so I don't know. I'm sure the
Ph.D.'s at the
companies who sell the "good" stuff have their point of view.


Michael Mead

__________________________________________________________________________________

Garry Burgess wrote:

} ------------------------------------------------------------------------
}
} The Microscopy ListServer -- Sponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- Send Email to
} ListServer-at-MSA.Microscopy.Com
} -----------------------------
} -----------------------------------------.
}
} First of all, I'd like to thank all of those who cared to respond to
} my
} question, especially Dr. Garber from SPI who gave a detailed and
} thoughtful response. And OK, I get the message, it's just not worth
} the
} risk.
}
} But I have another question that perhaps you people also might have
} some
} thoughts on. If I am understanding them properly, some of the
} Pathologists here claim that they can tell the difference between a
} specimen that was delayed before putting into Glut., resulting in
} artifact such as swollen mitochondria, vs. poor fixation as a result
} of
} outdated glutaraldehyde, such as damaged membranes in general. Is
} this
} sort of reasoning valid?
}
} Just curious,
} Garry



--------------CEF267DADC33931662890F41
Content-Type: text/html; charset=us-ascii
Content-Transfer-Encoding: 7bit

{HTML}
{FONT SIZE=+1} ----------------------------------------------------------------------- {/FONT}
{BR} {FONT SIZE=+1} The Microscopy ListServer -- Sponsor: The Microscopy
Society of America {/FONT}
{BR} {FONT SIZE=+1} To  Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {/FONT}
{BR} {FONT SIZE=+1} -----------------------------------------------------------------------. {/FONT} {FONT SIZE=+1} {/FONT}

{P} {FONT SIZE=+1} In response to Garry Burgess's question regarding swollen
mitochondria {/FONT}
{BR} {FONT SIZE=+1} sometimes referred to as "popcorn" mitochondria, the
only time I have seen {/FONT}
{BR} {FONT SIZE=+1} them perfectly preserved was after whole body perfusion
of fixative through {/FONT}
{BR} {FONT SIZE=+1} a living and pumping heart. {/FONT} {FONT SIZE=+1} {/FONT}

{P} {FONT SIZE=+1} Obviously, it is not likely that pathologists have tissue
preserved in this way {/FONT}
{BR} {FONT SIZE=+1} from living humans.  I guess for argument sake,
I would ask the pathologist {/FONT}
{BR} {FONT SIZE=+1} weather the swollen mitochondria and other membrane defects
could be the {/FONT}
{BR} {FONT SIZE=+1} result of the pathology and not poor fixation. {/FONT} {FONT SIZE=+1} {/FONT}

{P} {FONT SIZE=+1} On the other hand,  dead cells such as cuticle and
cortical cells in hair have {/FONT}
{BR} {FONT SIZE=+1} well defined cell membrane complexes that look well preserved
even if you {/FONT}
{BR} {FONT SIZE=+1} don't fix the hair--although cell organelles like mitochondria
are missing. {/FONT} {FONT SIZE=+1} {/FONT}

{P} {FONT SIZE=+1} For perspective,  I should tell you that I once worked
in the Anatomy {/FONT}
{BR} {FONT SIZE=+1} Department at Loma Linda University and the professor
there didn't believe {/FONT}
{BR} {FONT SIZE=+1} in purified glutaraldehye.  He used the 25% stuff
and kept it under the lab bench. {/FONT}
{BR} {FONT SIZE=+1} His belief was that the older it got the better....something
to do with the ratio of {/FONT}
{BR} {FONT SIZE=+1} dimer to trimer.  I'm not a chemist, so I don't
know.  I'm sure the Ph.D.'s at the {/FONT}
{BR} {FONT SIZE=+1} companies who sell the "good" stuff  have their
point of view. {/FONT}

{P}  
{BR} {FONT SIZE=+1} Michael Mead {/FONT}

{P} __________________________________________________________________________________
{BR} Garry Burgess wrote:
{BLOCKQUOTE TYPE=CITE} ------------------------------------------------------------------------
{BR} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

{P} First of all, I'd like to thank all of those who cared to respond to
my
{BR} question, especially Dr. Garber from SPI who gave a detailed and
{BR} thoughtful response.  And OK, I get the message, it's just not
worth the
{BR} risk.

{P} But I have another question that perhaps you people also might have
some
{BR} thoughts on.  If I am understanding them properly, some of the
{BR} Pathologists here claim that they can tell the difference between a
{BR} specimen that was delayed before putting into Glut., resulting in
{BR} artifact such as swollen mitochondria, vs. poor fixation as a result
of
{BR} outdated glutaraldehyde, such as damaged membranes in general. 
Is this
{BR} sort of reasoning valid?

{P} Just curious,
{BR} Garry {/BLOCKQUOTE}
   {/HTML}

--------------CEF267DADC33931662890F41--





From: Deepak Edward :      deepedwa-at-uic.edu
Date: Sat, 19 Jul 1997 13:53:22 -0500
Subject: Use of Pixera digital camera system

Contents Retrieved from Microscopy Listserver Archives
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Hello everbody,


Does anybody out there have experience with the {bold} Pixera digital
camera system {/bold} . Would appreciate your comments on the image
quality, the possibility of converting the images to publication quality
prints on a good printer and the level of technical support provided by
the company.


Thanks for your help


Deepak Edward






From: oshel-at-ux1.cso.uiuc.edu (Philip Oshel)
Date: Sat, 19 Jul 1997 19:44:40 -0500
Subject: apologies

Contents Retrieved from Microscopy Listserver Archives
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My apologies to the list. I sent a private email that seems to have gotten
posted to the MDS mail server. (And Eudora is configured not to do that ...
the net gremlins at work?)

Phil

****be famous! send in a tech tip or question***
Philip Oshel
Technical Editor, Microscopy Today
Station A
PO Box 5037
Champaign, IL 61825-5037
oshel-at-ux1.cso.uiuc.edu








From: Goldmarker-at-aol.com
Date: Sun, 20 Jul 1997 08:27:51 -0400 (EDT)
Subject: Used Knifemaker!

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Colleagues -

Would anyone be interested in a used LKB Knifemaker (7800) with cover + six
boxes of LKB glass? Fred says it is in excellent condition and that the
price would be in the $2500-$3000 range.

If interested, please contact Fred directly at fredl-at-awod.com (Fred G.
Lightfoot)
or call Fred at (803) 856-8613.

Thanks and regards, Don Cox, Goldmark Biologicals, goldmarker-at-aol.com







From: Goldmarker-at-aol.com
Date: Sun, 20 Jul 1997 08:36:23 -0400 (EDT)
Subject: Used Knifemaker!

Contents Retrieved from Microscopy Listserver Archives
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Dear Colleagues -

Would anyone have an interest in purchasing a used LKB Knifemaker (7800) with
six boxes of LKB glass? I think the price will be in the $2500 range. Fred
says it is in excellent condition.

If interested, please contact Fred Lightfoot at

fredl-at-awod.com (Fred G. Lightfoot)

or call Fred at (803) 856-8613

Regards, Don Cox
Goldmark Biologicals
goldmarker-at-aol.com




From: P.V.Hatton :      P.V.Hatton-at-sheffield.ac.uk
Date: Sun, 20 Jul 1997 18:33:48 +0100
Subject: Decalcification for TEM

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We section hard tissues containing synthetic biomaterials. While we
usually follow a conventional EDTA decal. route, we also sometimes
apply a little EDTA to the block face (having embedded in LR White)
to decal. "in situ" while sectioning. This alternative seems to
work, but as we are using diamond knives it may be purely
psychological! I have never seen it written up.

Best wishes, Paul


Dr Paul V. Hatton
Lecturer in Biomaterials
School of Clinical Dentistry
University of Sheffield
Claremont Crescent
SHEFFIELD S10 2TA

Tel. (0114) 271 7938
Fax. (0114) 2665326
or 2797050




From: Hong Yi :      hyi-at-emory.edu
Date: Sun, 20 Jul 1997 15:32:37 -0400 (EDT)
Subject: Microtome

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Dear microscopists:

We have a MT5000 Sorvall Ultra Microtome in storage. It is in
very good condition. Those who are interested, please email me at
hyi-at-emory.edu

Hong Yi
hyi-at-emory.edu





From: azita ariapour :      azita.ariapour-at-utoronto.ca
Date: Fri, 18 Jul 1997 14:19:00 -0400
Subject: unsubscribe

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From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Fri, 18 Jul 1997 10:24:16 -0500
Subject: Expired Glut. Another Question

Contents Retrieved from Microscopy Listserver Archives
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First of all, I'd like to thank all of those who cared to respond to my
question, especially Dr. Garber from SPI who gave a detailed and
thoughtful response. And OK, I get the message, it's just not worth the
risk.

But I have another question that perhaps you people also might have some
thoughts on. If I am understanding them properly, some of the
Pathologists here claim that they can tell the difference between a
specimen that was delayed before putting into Glut., resulting in
artifact such as swollen mitochondria, vs. poor fixation as a result of
outdated glutaraldehyde, such as damaged membranes in general. Is this
sort of reasoning valid?

Just curious,
Garry





From: Woody.N.White-at-mcdermott.com
Date: 7/17/97 7:12 AM
Subject: Polaron sputter target

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For an inexpensive target: Obtain a piece of OFHC Cu foil, cut
circle slightly larger than target, unscrew the existing target,
fold/crimp the foil over the existing target. Even if you have to get
the foil from Alfa/Aesar or the like, it should cost only about
$100, compared to the several hundred+ vendors want for a
"real" target.

Will be curious how well you are able to sputter Cu with the Polaron.

Woody White
Mcdermott Technology, Inc.

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_________________________________


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

Hello everyone,

Can anyone recommend a vendor to supply a Cu target for a Polaron E5000
sputter coater?
Thanks.

Owen


Owen P. Mills
Michigan Technological University
Metallurgical & Materials Engineering
Rm 512 MME Building
Houghton, MI 49931
906-487-2002
906-487-2934 FAX
opmills-at-mtu.edu




From: jeharper-at-amoco.com
Date: 7/14/97 2:02 PM
Subject: RE: Rush Lab + Projection Slides

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In my previous life doing TEM we would develop and wash negatives for
about 5 minutes, then rinse in methanol. Air dry or even a blow dryer
on low got us a dry negative in 15-20 minutes. I seem to remember
that there was a little clouding of the negative and you can't get too
anxious with the hair dryer.

Now, with negative, flatbed scanners one could scan the negative (10
minutes) and print an image on transparency paper with an inkjet
(Epson 1440 dpi or a dyesub printer). Haven't tried this with TEM
negatives but I know people who do.

Seems like about one hour would do it.

What about a positive TEM film that you could project directly.




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

Randy,

Sometimes when I want to make a transparency for seminars, or workshops
or whatever, I put the EM negative into the enlarger and project it to a
small 2X2" size on to yet another piece of EM film that I've cut in half
for this purpose. (you need a special enlarger lens to get this small!).
Then I cut it to size and mount it in a glass slide mount, and voila, I
have supersize black and white slides, with good resolution and
contrast. (at least better than 35mm projection slides) But sometimes
in the past, if I tried to rush things, I notice that the image turned
brown. To fix this problem though, I simply re-fix and re-wash this
image, and the brown discoloration (which is probably some residual
silver halide and fix) is removed, and all is well again.

Garry

} ----------
} From: rtind-at-siu.edu[SMTP:rtind-at-siu.edu]
} Sent: 14 July, 1997 14:51
} To: Garry Burgess
} Subject: RE: Rush Lab
}
} } ------------------------------------------------------------------------
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America





From: Mark Tashis :      mark_t-at-cc.huji.ac.il
Date: Mon, 21 Jul 1997 08:40:17 -0700
Subject: unsubscribe

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Message-ID: {33D382E1.13F9-at-cc.huji.ac.il}

unsubscribe




From: Bennett, Cynthia, HDG / FHF :      bennett-at-msmhdg.hoechst.com
Date: Mon, 21 Jul 1997 08:58:00 +0200
Subject: plasma etching units

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Hello folks,

Thanks ever so much for all those comments on "burn" marks. As a newbie,
I'm grateful for your help. Now I've got a question on sample
preparation.

We're currently doing many different types of sample preparation in an
"inherited" Edwards Auto 306, which works. But because we do so many
different things in it, we're always having to rearrange its "innards"
and this is turning out to be a bottleneck. So I was wondering about
doing some of the preparations in a separate unit.

In particular, I'm interested in purchasing equipment for plasma etching
and carbon coating. (IF--or WHEN-- we can afford it, that is!) So I'm
interested in hearing peoples' recommendations, good and bad experiences
and so forth. I also have no idea (yet) how much these things cost.

We do the etching to get a better look at those small inorganic
particles embedded in a polymer matrix that I mentioned in the EDX spot
mode conversation.

(And if vendors would like to contact me, you are welcome to do so!
Please do this directly and not via the listserver.)

Cindy Bennett

****************************************************
Dr. Cynthia Bennett
Hoechst Diafoil GmbH
Postfach 3365
D-55232 Wiesbaden
Germany
bennett-at-msmhdg.hoechst.com
tel.: +49-611-962-8123
fax: +49-611-962-9413

disclaimer: All opinions expressed are my own and not necessarily those
of my employer.




From: Hard, Robert :      rhard-at-ubmed.buffalo.edu
Date: Mon, 21 Jul 97 10:32:00 PDT
Subject: Course Announcement

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Course Announcement

Title: Optical Microscopy and Imaging in the Biomedical Sciences

When: October 8 - October 16, 1997

Where: Marine Biology Laboratory, Woods Hole, MA, USA

Tuition: $1950 (Includes room and board)

Application Deadline: August 5, 1997

Admission application and information:
Carol Harnel, Admissions Coordinator
Marine Biological Laboratory
7 MBL Street
Woods Hole, MA 02543-1015
(508) 289-7401
Internet: admissions-at-mbl.edu
WWW: http://www.mbl.edu

Course Director: Colin S. Izzard, State University of New York -at- Albany
Phone: [518] 442 - 4367
EMail: csizzard-at-csc.albany.edu

Course Description:

For Whom:
Designed primarily for research scientists, physicians, postdoctoral
trainees and advanced graduate students in animal, plant, medical and
material sciences. Non-biologists seeking a comprehensive introduction to
microscopy and video-imaging will benefit greatly from this course as well.
There are no specific prerequisites, but an understanding of the basic
principles of optics is desirable. Limited to 24 students.

The eight day course consists of lectures, laboratory demonstrations,
exercises and discussions that will enable the participant to obtain and
interpret microscope images of high quality, to perform quantitative optical
measurements, and to produce photographic and video records for documentation
and analysis.

Topics to be covered include:
principles of microscope design and image formation
bright field, dark field, phase contrast, differential
interference contrast, interference reflection, and
fluorescence microscopy
confocal scanning microscopy and image deconvolution
digital image restoration and 3-D reconstruction
video imaging, recording, enhancement, and intensification
analog and digital image processing and analysis
fluorescent probes and ratio-imaging
laser tweezers and laser scissors

Applications to live cells will be emphasized; other specimens will be
covered as well.

Students will have direct hands-on experience with state-of-the-art
microscopes, video cameras, recorders and image processing equipment provided
by major optical and electronics companies. Instruction will be provided by
experienced staff from universities and industry.

Students are encouraged to bring their own biological (primary
cultures, cell lines, etc.) and material specimens and to discuss individual
research problems with the faculty.





From: :      kna101-at-utdallas.edu
Date: Mon, 21 Jul 1997 08:38:59 -0500 (CDT)
Subject: Re: Decalcification for TEM

Contents Retrieved from Microscopy Listserver Archives
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Paul,

I've seen someone use this technique to decalcify and section
tissue in epon years ago. It worked fine. I also don't know where any
written info can be found about it.

By epon, I mean specifically medcast from Ted Pella Inc., I
understand this particular media has been discontinued, but I don't think
the EDTA decalcification is limited by the media. It only decalcified a
few mm-s of surface tissue.

Karen Pawlowski
Lab Tech
UT Southwestern Med. Ctr.
Student/ UT Dallas

On Sun, 20 Jul 1997, P.V.Hatton wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} We section hard tissues containing synthetic biomaterials. While we
} usually follow a conventional EDTA decal. route, we also sometimes
} apply a little EDTA to the block face (having embedded in LR White)
} to decal. "in situ" while sectioning. This alternative seems to
} work, but as we are using diamond knives it may be purely
} psychological! I have never seen it written up.
}
} Best wishes, Paul
}
}
} Dr Paul V. Hatton
} Lecturer in Biomaterials
} School of Clinical Dentistry
} University of Sheffield
} Claremont Crescent
} SHEFFIELD S10 2TA
}
} Tel. (0114) 271 7938
} Fax. (0114) 2665326
} or 2797050
}





From: Robert H. Olley :      R.H.Olley-at-reading.ac.uk
Date: Mon, 21 Jul 1997 14:19:29 +0100 (BST)
Subject: Etching with gas

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This is prompted by Cynthia Bennett's recent request in regard to plasma
etching units. A few years ago, we were briefly interested in such
things, particularly in how to remove the surface from a polymeric
material without damaging the underlying substructure. However, for our
purposes we found that plasma etching, ion mills, etc., would be far too
destructive - not only would there be heating problems, but also all
those ions running wild would tend to cause a lot of chemical damage.

We had just got round to trying out ATOMIC OXYGEN, generated in a radio
frequency discharge, and the first result or two seemed promising, and
then with a great bureaucratic reshuffle the owner of the equipmment
pulled up sticks and went elsewhere. Does anyone know where such
equipment might be obtainable at reasonable cost?

If anyone has experience with this sort of thing, I would be pleased to
hear from them.

Thanks in advance,

+------------------------------------------------------------------------+
| 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: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Mon, 21 Jul 1997 10:13:14 -0500
Subject: Optimal Glut. Concentration

Contents Retrieved from Microscopy Listserver Archives
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Now that this whole question of expired Glut. and fixation has come up,
I'm wondering if perhaps my 2.5% solution that we use here is perhaps
suboptimal, and whether or not we might be better off using a strong
glutaraldehyde concentration such as 4% for our routine fixation.

Is there anyone else out there fixing human tissue routinely? I would
be interested in what concentration of glutaraldehyde that you people
are using.

Garry




From: Garry Burgess :      GBurgess-at-exchange.hsc.mb.ca
Date: Mon, 21 Jul 1997 10:13:14 -0500
Subject: Optimal Glut. Concentration

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

Now that this whole question of expired Glut. and fixation has come up,
I'm wondering if perhaps my 2.5% solution that we use here is perhaps
suboptimal, and whether or not we might be better off using a strong
glutaraldehyde concentration such as 4% for our routine fixation.

Is there anyone else out there fixing human tissue routinely? I would
be interested in what concentration of glutaraldehyde that you people
are using.

Garry




From: Dr. David C. Bell :      dcb-at-MIT.EDU
Date: Mon, 21 Jul 1997 12:05:06 -0400
Subject: Canada Balsam

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Hello all,
I was just asked for answers regarding Canada Balsam, which
I hope some member of the group could assist me with, they are..

Questions regarding Canada Balsam,
1) When was it first used for microscopy?
2) How long does it last before degrading?
3) What if any are the aging effects?
4) Does it interfere or affect the sample in any way?

The question was actually asked for a different application
other than microscopy, but I thought it would be an interesting
discussion none the less.

Thanks

David
Dr. David C. Bell
Room 13-1018 E-Mail: dcb-at-MIT.EDU
Center for Mat. Sci. and Eng. PH: (617) 253-3317
Massachusetts Institute of Technology FAX: (617) 258-6478
77 Massachusetts Ave, Cambridge, MA 02139-4307




From: PESTO 224 STOLZENBERG :      Pesto-at-erols.com
Date: Mon, 21 Jul 1997 12:29:49 +0000
Subject: (no subject)

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We are looking for a used High-Voltage Tank for a JEOL l00.
Can anyone help us?
Please answer us direct. Thank you. Peter Stolzenberg
PESTO INC. pesto-at-erols.com




From: PESTO 224 STOLZENBERG :      Pesto-at-erols.com
Date: Mon, 21 Jul 1997 12:36:25 +0000
Subject: High Voltage Tank needed for JEOL

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To all:
Can anyone help us getting a used Jeol l00 tank. We would appreciate
any leads. Please E-Mail us direct. Thank you!
Peter Stolzenberg,Pesto Inc. P.O. Box 648, GWYNEDD VALLEY ,PA 19437
215-699-6160 FAX215-699-5275 E-Mail: pesto-at-erols.com




From: Michael Shaffer :      mshaf-at-OREGON.UOREGON.EDU
Date: Mon, 21 Jul 1997 09:35:23 -0700
Subject: EPMA: x-ray wavelength database

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I remember there exsisting a database of x-ray wavelengths ... does
anyone know from where it can be downloaded???

TIA & cheerios, shAf
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/}
Michael Shaffer, R.A. - University of Oregon Electron Probe Facility
mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu
http://darkwing.uoregon.edu/~mshaf/epmahome/




From: Vickie Frohlich :      vickie-at-vms2.macc.wisc.edu
Date: Mon, 21 Jul 1997 11:56:39 -0600
Subject: An Affordable Symposium

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To anyone wishing to attend the pre-MSA symposium on "Applications of
Multiple Photon Excitation Imaging"


Limited space is currently available for the Symposium. Please
register in advance. We may not be able to take registrations at the
door.


Registration for the pre-MSA shortcourse is closed. Thank you to all
who enquired about attending.

{fontfamily} {param} Courier {/param} {bigger}

"APPLICATIONS OF MULTI-PHOTON EXCITATION IMAGING"


at the=20


SHERATON CITY CENTER, Cleveland, Ohio


August 9 and 10, 1997


Registration Fee: $ 30.00 (Symposium only)



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

SYMPOSIUM REGISTRATION FORM


"APPLICATIONS OF MULTI-PHOTON EXCITATION IMAGING"


NAME: SS#:

ADDRESS:

CITY: STATE: ZIP:

PHONE: FAX:

EMAIL:


REGISTRATION FEE: $ 30.00


Check to MSA enclosed:=20
{/bigger} {/fontfamily} {bigger} {fontfamily} {param} New_York {/param} =03 {/fontfa=
mily} {fontfamily} {param} Courier {/param}


Credit Card Number (Visa or Master Card only): =20


_ _ _ _ - _ _ _ _ - _ _ _ _ - _ _ _ _


Expires: Month _ _ Year _ _


Signature:______________________________________


Print Name on Card:_____________________________


MAIL TO: Dawn Volkman

Integrated Microscopy Resource

University of Wisconsin

Madison, WI 53706=20

Phone: (608) 265-3083

Fax: (608) 265-4076

dvolkman-at-students.wisc.edu

www.bocklabs.wisc.edu/imr/home.htm

(Note New Web Address)


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

{/fontfamily} {/bigger} {fontfamily} {param} Courier {/param} Sponsored by:

Integrated Microscopy Resource

University Wisconsin-Madison


Center for Light Microscope

Imaging and Biotechnology=20

Carnegie Mellon University


and=20


Microscopy Society of America



SYMPOSIUM PROGRAM:

SATURDAY 9 AUG 97 - Sheraton Hotel Conference room

8:00- 8:50 Winfried Denk - Lucent Technology/Bell Labs

"Multi-Photon Excitation: From the Beginnings to

Applications in Neuroscience"

8:50- 9:30 Warren Zipfel - Cornell University

"Multi-Photon Excitation of Intrinsic Fluorescence

in Cells and Intact Tissue"

9:30-10:10 David Piston - Vanderbilt University

"Metabolism, Development, and Two-Photon=20

Excitation Microscopy"

10:10-10:30 Coffee Break

10:30-11:10 Stefan Hell - University of Turku, Finland

"Light Microscopy with Sub-100 nm Resolution=20

in Three Dimensions"

11:10-11:50 Enrico Gratton - University of Illinois

"Two-Photon Fluctuation Correlation Spectroscopy"

11:50-12:30 John White - University of Wisconsin

"Multi-Photon Excitation Imaging Applied

to the Study of Developing Embryos"

12:30- 1:30 Lunch

1:30- 2:10 Ursula Keller - ETH, Switzerland

"SESAM Devices for Passive Pulse Generation

from 6.5 fs to ns in Solid-State Lasers"=20

2:10- 2:50 Frank Wise - Cornell University

"Femtosecond-Pulse Sources for Nonlinear

Laser Microscopy"

2:50- 3:10 Coffee Break

3:10- 3:50 Allister Ferguson - University of Glasgow, UK

"User-Friendly Femtosecond Sources for Multi-Photon

Fluorescence Imaging"

3:50- 5:30 Question/answer session with Saturday speakers

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

SUNDAY 10 AUG 97 - Sheraton Hotel Conference room

8:00- 8:30 Brad Amos - Medical Research Council, UK

"MPEFM: a YLF Laser Applied to Bioimaging"

8:30- 9:00 Hans Gerritsen - Univ. Utrecht, Netherlands

"Fluorescence Lifetime Contrast in Two-photon=20

Excitation Microscopy"

9:00- 9:30 Rafael Yuste - Columbia University

"Two-photon Imaging of Dendritic Spines"

9:30-10:00 Rebecca Williams - Cornell University

"Three-photon Excited Fluorescence Microscopy=20

of Serotonin Release"

10:00-10:15 Coffee Break

10:15-10:45 Steve Potter - California Institute of Technology

"3D Time-lapse Imaging of Hippocampal Slices"

10:45-11:15 Jackie Schiller - Mayo Clinic

"Multi-photon Excitation Uncaging in Rat Brain

Slices"

11:15-11:45 Phil Hockberber - Northwestern University

"Phototoxicity: Avoidance Using 2PEFM" =20

11:45-12:15 Mark Cannell - University of London, UK

"2PEFM of Calcium in Muscle Cells"

12:15-12:45 Martin Kohler - Karolinska Hospital, Sweden

"2PEFM of Calcium in Pancreatic Beta Cells"

"Islet of Langerhans, Calcium, and Two-photon

Excitation microscopy"

12:45- 1:30 LUNCH

1:30- 4:30 Talks by reps from commercial exhibitors:

Biorad, Nikon, Spectra-Physics, Coherent etc.

=20

4:30- 5:30 Question/answer session with Saturday speakers

{/fontfamily}






From: Larry Stoter :      LPS-at-teknesis.demon.co.uk
Date: Sat, 19 Jul 1997 09:28:22 +0100
Subject: Re: Low-dose for Philips420

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} Dear Microscopists,
} we plann to use TEM Philips 420 for Low-dose work. We will need Low-dose
} system for this microscope. Unfortunately Philips said us,that they didn't
} produce Low-dose for this kind of microscope,already.
} Can you help us please, how or where we could get this Low-dose unit or
} how to work in low-dose conditions /to prevent destroing samples during
} focusing/ without this system on TEM Philips 420 ?
} All your responces or opinions will wery useful for us.
}
} Thank You very much
} Milos
}
} Milos Motejl
} Lab. of Biomembranes
} South Bohemian University
} Ceske Budejovice
} Czech Republic
}
} motejl-at-paru.cas.cz
} tel. 042-038/7775485
} ................................

Philips used to produce free-standing Low Dose modules for their 400 series
TEMs - off hand, I don't recall the part number, but if you check around
you might find a used one available.

Actually, the easiest approach to low dose is the simplest - work at low
magnification. I used to be an applications specialist for Philips, and had
the time to experiment with different ways of using a TEM. With a little
pre-calibration and experiment, I could repeatedly, with a single
micrograph, record the 0.9 nm lattice spacing of crocidolite with the TEM
at 10,000 x magnification - the point being that you can record quite high
resolution images at low magnification, which gives you sufficient
intensity at the film to record the image but considerably reduces the dose
to specimen, compared with operating at high mag. Note, that this approach
fails if you take the mag into the LM mag range, as the whole optics of the
microscope changes at this point, one consequence being a drastic drop in
resolution.

You can further reduce the dose to the specimen if you have a STEM unit.
Basically the idea is to operate the TEM column in a high mag/high res mode
and very low beam intensity but use the STEM unit to image the specimen at
low mag/low res, so as to locate a the region of interest.

With a standard STEM set up, you can only image a small part of the field
of view. However, if you have a computer connection to allow you to control
beam position, it is straightforward to produce a short routine that will
step the illuminated area across the whole area covered by the film. With
this approach, I have 'easily' recorded diffraction patterns and images
from parafin wax.

With the same sort of computer control set up, it is not much more
difficult to duplicate most of the functions of the low dose unit, except
that you can't change the illumination focus - but that can be done
manually at the appropriate step in the sequence.

Regards,
Larry Stoter






From: Tom Christensen :      tgc-at-bu.edu
Date: Mon, 21 Jul 1997 13:44:05 -0700
Subject: Re: Optimal Glut. Concentration

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Garry Burgess wrote:
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Now that this whole question of expired Glut. and fixation has come up,
} I'm wondering if perhaps my 2.5% solution that we use here is perhaps
} suboptimal, and whether or not we might be better off using a strong
} glutaraldehyde concentration such as 4% for our routine fixation.
}
} Is there anyone else out there fixing human tissue routinely? I would
} be interested in what concentration of glutaraldehyde that you people
} are using.
}
} Garry


Here in the EM Facility at Boston Medical Center, we use 2.5% glut in
0.2M cacodylate buffer for all human specimens; the fix lasts for
several months in the fridge and yields very high quality results if the
tissue is fixed promptly at the biopsy site. If a surgical specimen has
been held overnight in the cold and is then sampled for EM in the
morning, the preservation suffers but is still suitable for diagnosis.

Dr. Tom Christensen
Director, EM Facility
Boston Medical Center
Boston, Mass




From: Darrell Miles :      milesd-at-US.ibm.com
Date: Mon, 21 Jul 1997 15:35:48 -0400
Subject: Re: Questions on device failure analysis

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Hello,

I do not know if you have gotten any other replies on this. We do failure
analysis on integrated
circuits. There are two types of "short circuits". One is a resistive short,
and the other is a
leakage path where hole-electron recombination occurs. Resistive shorts can be
located
with liquid crystal methods that sense the thermal dissipation. (There are
other, more complex,
methods, but we use liquid crystal extensively) Leakage paths can be located
with systems
based on "night vision" technology that was developed for the military.
Hole-electron recom-
bination releases the excess energy as photons, and the light amplification
allows the imaging
of the emission site.

Darrell Miles
IBM Microelectronics
Test and Analytical Services
http://www.chips.ibm.com/services/asg




From: John Arnott :      ladres-at-worldnet.att.net
Date: Mon, 21 Jul 1997 15:34:00 -0400
Subject: Casting Sesssion at MSA '98

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To all who are interested,

A tentative okay has been given for a session on corrosion casting for
the 1998 meeting in Atlanta.
We would urge any of our Mercox users who are interested in presenting a
paper to contact us at Ladd Research or Dr. Fred Hossler at:

Dr. Fred Hossler
Professor of Anatomy
East Tennesse State University
Johnson City, TN 37614
e-mail SEMTEMman-at-aol.com

Methadology is of particular interest.

We would also like to have some opinions on the advantages/disadvantages
of using clear, blue or red mercox for casting.

Thank you for help,

John Arnott




From: Dr. David Hall :      hall-at-aecom.yu.edu
Date: Mon, 21 Jul 1997 16:08:52 -0400
Subject: histochemical stains for plastic sections

Contents Retrieved from Microscopy Listserver Archives
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In March 1997 we requested protocols and references for staining semi-thin
sections of vertebrate tissues embedded in plastic; our original standard
stain was toluidine blue.

We received 14 responses to this request, which we have edited to a ~7 page
file. Responses included stains such as hematoxalin, alcian blue, eosin,
Mayer's mucicarmine, methyl green, methylene blue/azure II, basic fuschin,
and Stevenel's blue. A copy of the file is available upon request by
e-mail. We have begun using a polychrome stain (see Van Reempts and
Borgers, 1975, Stain Tech. 50:19-23) with pleasing results.

Christine Roy and David Hall
Albert Einstein College of Medicine
Bronx, NY 10461




From: Steve Barlow :      sbarlow-at-sunstroke.sdsu.edu
Date: Mon, 21 Jul 1997 13:09:02 -0700
Subject: video board

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I need a Nubus video board for a Mac PPC7100 for video conferencing.
Anyone have a suggestion where I can get one?

thanks

steve

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

Dr. Steven Barlow
EM Facility/Biology Department
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/EM_Facility






From: Lifan Chen :      lfchen-at-zen.sb.fsu.edu
Date: Mon, 21 Jul 1997 16:21:20 -0400
Subject: E-mail address

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Hello all,

Does anybody know the e-mail address of Prof. P. Kruit who is the chief editor
of Ultramicroscopy? Somebody needs his email address.

Thanks a lot,

Lifan Chen




From: wise-at-vaxa.cis.uwosh.edu
Date: Mon, 21 Jul 1997 16:23:22 +0000
Subject: Film for SEM

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I am interested in finding out what people are using for 4"x5" SEM film. I
have used Polaroid Type 55 P/N and Kodak type 4427 Commercial Film. At
$1.50 to $2.00 per exposure, both are a little too expensive for student
use (i.e. low good photo to bad photo ratio). Can anyone recommend a less
expensive alternative? My predecessor used to buy 200 foot rolls of
surplus aerial photography film (4 or 5 inches wide) which he would cut to
the proper length. Final cost was less than $0.05 per sheet. Any similar
suggestions?

Bob


Dr. Robert R. Wise
Department of Biology
University of Wisconsin-Oshkosh
Oshkosh, WI 54901

(414) 424-3404 tel
(414) 424-1101 fax
wise-at-uwosh.edu

Note: area code will change to 920 on July 26th






From: Steven W. Miller :      Steven_W_Miller-at-CompuServe.COM
Date: Mon, 21 Jul 1997 18:32:58 -0400
Subject: SEM: sample preparation for particles dispersed in oil???

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Nora Pratta
Centro Regional de Investigacion y Desarrollo
Santa Fe - Argentina
Inquired about doing SEM/BSE/EDX on particulates in oil and oil/water
mixture.

If the oil or oil water mixture can be 'frozen' at liquid nitrogen
temperatures by rapid freezing in a metal block or propane jet freezer th=
en
transferred to a cryo ultramicrotome where you could prepare frozen
sections for examination on an SEM cold stage. =

This is a very expensive but technically superb method. An alternate woul=
d
be to freeze dry the particulates out of the section onto a carbon
substrate or other suitable substrate. You will lose some of the
distribution information in the X-Y plane but should resolve the Z
distribution represented by your sections themselves.

There are some commercial labs with this equipment and some private
companies that may collaborate if the subject interests them.

You may contact me off line or visit the Web Site: =


http://www.RMC-Scientific.com/microtomes/ =


We are a commercial manufacturer of all of the instruments listed above.=


Steve Miller
Director of Sales
RMC
3450 S. Broadmont, Suite 100
Tucson, AZ 85713
Tel 520-903-9366
Fax 520-903-0132



=

=





From: Stephen A. Shaffer :      sshaffer-at-microdataware.com
Date: Mon, 21 Jul 1997 16:45:54 -0700
Subject: Inter/Micro 97 Day One

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Inter/Micro 97 - Day One

"Inter/Micro" is a meeting, now in its 49th year, held annually in
Chicago and hosted by the McCrone Research Institute (McRI). I am
attending the meeting as I have almost every year for the past 25 or
so. However it is a meeting little known to those without regular
contact with McRI. As a way of introducing this under-attended meeting
to a wider audience I thought the readers of this list might find it
informative to have a summary of the technical highlights and other
goings-on at Inter/Micro. If I can, I'll write a daily summary every
evening and post it to this list. Otherwise I'll just summarize and
post as time permits. I'll mention two or three of those papers I found
most interesting each day.

This year approximately 60 technical presentations are scheduled. A
single meeting room is used and there are no parallel sessions. One has
an opportunity to hear all of the papers and I find it quite valuable to
sit in on papers which might not be in an area of my own personal
focus. This "cross-fertilization" of ideas and techniques is often the
most enlightening thing that I experience at technical meetings and I am
glad they've continued the tradition at Inter/Micro. There will also be
an exhibition of products and equipment, as is usual at such meetings.
A full program is available at McCrone's web page, http://www.mcri.org.
I won't duplicate that here but I'll just mention that the themes for
the various sessions for the week are as follows:
Monday: General Microscopy
Tuesday am: Instrumentation
Tuesday pm: Techniques
Wednesday: History and Art
Wednesday Eve: Dinner in Conjunction with the State
Microscopical Society of Illinois (SMSI)
Thursday: Forensic Microscopy
Friday: A Tutorial on Dispersion Staining

The highlights (for me) of Monday's program included the following.

The program was opened with a talk by Brian Ford on "Crytosporidium, a
New Threat from Water Supplies." Brian discussed how Cryptosporidium is
representative of a "new" class of microorganisms threatening our modern
society. Of course, it is not new at all however a combination of
factors is working together to make many existing organisms newly
hazardous. These factors include new practices and factors coming with
technological advances such as the wearing of contact lenses which
provide a previously non-existing environment in which certain organisms
can thrive. Another factor is the emerging resistance of some organisms
to existing treatments. Previously "eradicated" threats are reemerging
as "new" threats again. In the case of Cryptosporidium, we have an
organism causing severe but usually non-fatal intestinal distress that
we are immune from after the initial exposure and bout of sickness. But
it has become so "rare" as a contaminant that few of us received the
immunizing infection early in our lives, leaving large segments of the
population subject to infection when water treatments fail or other
sources of the protozoa present to the population.

Another highlight today was a paper by John Wuepper of Whirlpool
entitled "Problem Solving via Analytical Microscopy: What is it Really
Worth?" At Inter/Micro we have on many occations over the years
lamented the under-appreciated and under-valued status of the work we do
on behalf of industry and society at large. John suggested an excellent
technique that we can use if we revise our thinking and the presentation
of our work to the corporations or agencies we serve. He demonstrated
with several examples that the "leverage" obtained from an investment in
microscopical problem solving is often enormous, in the range of 25 to
5,000 in the examples he gave. By "leverage" he meant the ratio of the
cost of the process to the savings enjoyed by the company as a result of
the work done. The "investments" may range from a few hundred dollars
to a hundred thousand or more, but the return and leverage is enormous.
In one instance he cited microscopy saved a company more than
$500,000,000 dollars. (Those figures will catch the eye of even the
most myopic bean-counter - my editorial, not John's!) A lively
discussion followed John's presentation and it is clear that his
approach of communicating with our host-employers in terms they
understand and respond to is critical to the success and growth,
sometimes just the survival, of a microscopy laboratory.

Jan Hinsch or Leica gave one of the most beautifully illustrated as well
as educational talks of the day when he spoke on "What Pleurosigma can
Tell the Microscopist." Pleurosigma Angulatum, a species of diatom, has
long been used as a microscope test object for evaluating the quality of
higher numerical aperture objectives. Jan's talk was one of those
wonderful half-hours where an audience gets to enjoy not only an
aestheticly beautiful talk but one that, through the instructional
insight of the author, makes crystal clear some technically difficult
fundamental principles. In the case of today's talk, those principles
dealt with the theoretical resolution limits of the light microscope and
I, along with many others in the audience I think, came away with with a
vastly better understanding of what had heretofor been a baffling
subject.

Another fascinating talk today illustrated the diversity of topics we
regularly enjoy at Inter/Micro. Ryan D. Tweney of the Department of
Psychology at Bowling Green State University spoke on "Cognition and the
Microscope." Ryan discussed and illustrated many of those murky
processes that stand between knowledge or observation on the one hand
and understanding on the other. It was apparent that he was only able
to scratch the surface of a subject which we would all benefit from
knowing much more about and I, for one, hope we'll see him back
regularly in the future.

I'll try to come back tomorrow with another update! Right now I've got
friendships to rekindle and think I hear the calling of a lonely brew
with my name on it.

Steve Shaffer
MicroDataware




From: wesleysm-at-biology.und.ac.za (James Wesley-Smith)
Date: Tue, 22 Jul 1997 08:02:38 +-200
Subject: LM mountant. Summary

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Dear listeservers

Thank you to those of you who replied regarding LM mounting medium. It
appears as if oxidants in the medium are the main culprits. I am listing
these replies below.

One point worth mentioning about the common use of epoxy resins as a
coverslip mountant is that its refractive index is not 1.5 (I'm not sure of
its exact value). One may be able to get away with it working at low
numerical apertures, but it will take its toll at 0.65 and above.

Thanks again!

James Wesley-Smith
EM Unit
University of Natal
Durban, South Africa


I learned a trick from a DuPont-Sorvall rep (that tells you how long ago
that was!) that retards or eliminates ALL fading caused by oxidants in the
mounting medium. Add 1-2% BHT (the preservative used in bologna, hot dogs,
etc.) to any mounting medium. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
I have been using ENTELLAN which has ND20 1.49-1.5 and gives
excellent preservation of toluidine blue stained plant tissue for at least
4 years. I get it from Electron Microscopy Sciences who have a good list
of mounting media with refractive indicies in their catalogue.
-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
Hello
fading is the problem. We use these toluidine blue sections for histology
courses and we haven't find yet the no-fade mounting medium. Our best
choice is DEPEX, manufactured by GURR. Avoid EUKITT, fading occurrs in a
matter of hours. Some collegues have used cured epon, but it is a time
consuming process and fading does occur.
-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
I think the fading is caused by oxygen (which very
easily diffuses through hydrophobic media like resins).
We routinely leave such preparations uncovered, and add a drop of immersion
oil and coverslip to photograph. This prep is easily soaked off in xylene
to restain if necessary.
-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
I don't know about the refractive index, but I've used Epoxy resin as a
mountant quite successfully. It doesn't seem to fade Toluidine Blue
semi-thins.

DePeX is not too bad but I have had some fading over long periods.
-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-

I have been working with plant material embedded in Spurr resin for many,
many years. One micron sections were stained with toulidin blue or other
specific stains and permanent mounted with Permount, Fisher Scientific, and
no fading for decades. I have not seen any fading using DePeX, but Spurr
resin sections usually get very wrinkeled.

-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
We use the following protocol for toluidine blue (TB) staining
and permanent mounting:
Frozen or dewaxed abd hydrated paraffin sections
0.1% TB in acetate or phosphate buffer (generally at pH 2-3 for
sulfated glycosaminoglycans) 5 min
Rinsing in buffer
Precipitation with a 6:1 mixture of 2% aqueous KI and Kferricyanide
2-3 min
Mount with a drop of 25% aqueous gum arabic containing 2% fructose
without coverslip! Form a layer of te mounting medium using a
glass rod. Let the gum arabic layer dry at room temperature in
horizontal position (it takes generally one night). The
refractive index of the dried gum arabic is practically
identical to that of the glass.
Mount in DPX or Canada using a coverslip.
This procedure is good to produce permanent metachromatic staining.
Dimethylmethylene blue (DMMB) is a better metachromatic dye (Aldrich
Co, or SERVA)
The protocol is similar, except the poststaining stabilization. For
this purpose, 2% aqueous ammoniummolybdenate is used.
DMMB is a very strong metachromatic staining. It is very useful for
mast cells, cartilage, sulfomucins. In many cases, we use it
successfully in 0.05 or 0.01% aqueous solution for 5-10 min.

For more inforations, see Modis, L.: Organization os the
Extracellular Matrix: A Polarization Microscopic Approach. CRC Press,
Boca Raton, 1991. Chapter 12.






From: wesleysm-at-biology.und.ac.za (James Wesley-Smith)
Date: Tue, 22 Jul 1997 08:44:02 +-200
Subject: Specimen processing queries

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Kindly forgive me if these references have already been posted, but the
answers to many of these processing protocol queries can also be found in a
series of articles by Coetzee and van der Merwe, viz.

J Coetzee and CFvan der Merwe (1984) Extraction of substances during
glutaraldehyde fixation of plant cells. Journal of Microscopy 135, Pt2, pp
147-158.

J Coetzee and CFvan der Merwe (1985) Penetration rate of glutaraldehyde in
various buffers into plant tissue and gelatin gels. Journal of Microscopy
137, Pt2, pp 129-136.

J Coetzee and CFvan der Merwe (1986) The influence of processing protocol
on the ultrastructure of bean leaf cells. South African Journal of Botany,
52, pp 95-99.


These articles are compulsory reading for our trainee microscopists, since
they dispell many processing 'myths'.


James Wesley-Smith
EM Unit
University of Natal
Durban, South Africa







From: kothe-at-mwald5.chemie.uni-mainz.de
Date: Tue, 22 Jul 1997 13:13:37 +0200
Subject: Programs for analyzation of ED-patterns

Contents Retrieved from Microscopy Listserver Archives
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We want to analyse the intensity of ED-patterns of organic specimen. For
this we used ELD, a program which is included in the CRISP packages from
Calidris. Now we want to compare the intensity data from ELD with the
intensity getting from other programms. Does anybody knows some programs,
which we can used for intensity estimation? Can you tell something about
prices and the possibility to get such programms!

Hans Kothe
Working group Dr. Voigt-Martin
Universit=E4t Mainz
=20





From: Kevin Mackenzie :      k.s.mackenzie-at-abdn.ac.uk
Date: Tue, 22 Jul 1997 13:02:21 +0100 (BST)
Subject: 25th Scottish Microscopy Symposium

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25th Scottish Microscopy Group Symposium (First Circular)

Stakis Dunblane Hotel, Dunblane.
Wednesday 12 November 1997.


This the SILVER Scottish Microscopy Symposium will take place at the=20
above venue and the Organising Committee have arranged a Scientific=20
Programme which we hope will appeal to as many microscopists as=20
possible. Also a celebratory meal will mark this anniversary.

The following topics have been selected:

Environmental Scanning Electron Microscopy - Dirk van der Vall,=20
Eindhoven, Holland.

Advances in Confocal Microscopy - Tony Wilson, Oxford, England

Stereology - Vyvyan Howard, Liverpool, England

Cryo/Immunocytochemistry - Jeremy Skepper, Cambridge, England

These invited talks will be interspersed with short presentations. We=20
would welcome offers of short (10-15 minute) talks which deal with any=20
aspect of microscopy and in particular electron microscopy. There is an=20
abundance of useful techniques and protocols in daily use; if you think=20
that you have something which others could adopt or benefit from,=20
please send us your name and a brief title for your presentation to Ian=20
Roberts at {irober-at-scri.sari.ac.uk}

These meetings are enjoyable, interesting and useful and an=20
opportunity to meet and share ideas with fellow microscopists. The cost=20
will be =A320 (pounds).

Celebratory Dinner (evening)

To mark the 25th Anniversary of these meetings, we hope to arrange an=20
evening dinner to which all delegates and partners are invited to attend.=
=20
The separate cost of this will be =A325.00/head, and a favourable rate of=
=20
=A3100 per night/ per couple for dinner, bed and breakfast at the hotel has=
=20
been negotiated. If you wish to attend this evening function, please=20
contact Martin Maxwell at {MARTIN.MAXWELL-at-BBSRC.AC.UK}

A second circular will be sent in the future given details of all talks. Al=
so=20
there is a web page at http://www.abdn.ac.uk/~nhi691/smg97.htm that=20
gives more information.



Kevin Mackenzie
Tillydrone E.M. Unit
University of Aberdeen
Tillydrone Avenue
Aberdeen
AB9 2NT

Tel 01224-272847
Fax 01224-272396
Web site- http://www.abdn.ac.uk/~nhi691/


----------------------
Kevin Mackenzie
k.s.mackenzie-at-abdn.ac.uk







From: Dan Hill :      dh2-at-mole.bio.cam.ac.uk
Date: Tue, 22 Jul 1997 14:43:20 +0100 (BST)
Subject: TEM AEI EM801 to good home

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We have a working AEI EM801 TEM (1969) that is free to a good home,
otherwise its for the skip (valve technology but it does take 6 grids at a
time)

Contact

Dan Hill
Biochemistry Department
Cambridge University
Tennis Court Road
Cambridge
Cambs
CB2 1QW
UK

Tel 01223 333685
e-mail dh2-at-mole.bio.cam.ac.uk






From: rtind-at-siu.edu (Randy Tindall)
Date: Tue, 22 Jul 1997 09:36:44 -0600
Subject: Film for SEM

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We used to commonly use a Kodak film called Ektapan in 4x5 size. It is
developed in D-76. usually, although other standard film developers could
also be used. The advantage is that it's much cheaper than Polaroid PN 55,
at about $20 per 25-sheet box (last time we checked the price). The
disadvantages, of course, are that a developing set-up and darkroom are
required and you don't get an automatic study print.

Actually, it would seem that any common 4x5-inch film could be adapted to
SEM use, depending upon contrast requirements. T-Max 100 or 400, Plus-X
Pan, Ilford, or Agfa films, etc., all should work. All of these films
should be readily available and all use a variety of common developers.

Hope this helps.

Randy Tindall
Center for Electron Microscopy
Southern Illinois University at Carbondale






From: Michael Shaffer :      mshaf-at-OREGON.UOREGON.EDU
Date: Tue, 22 Jul 1997 08:27:25 -0700
Subject: EPMA: x-ray wavelength database

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Since my initial request for the x-ray wavelengths file I have since
found a Microsoft Access database file which was originally provided by
John Donovan (UC Berkeley). I have also received request for posting the
file for FTP (... altho one reply indicated the original "Fiori" file was
available at ftp://www.anc.anl.gov ...), and the John has since indicated
his database which also includes higher order lines is generally available.
I can make John's MDB file and my Excel (XLS v2.1) file available via
"anonymous" FTP. My XLS file is only slightly different, i.e., it has been
sorted by "Z" and "N" ...
Lastly, a word about the FTP site. It is actually a magneto-optical
drive and used for archiving image files. As a MO drive it will offer these
specific files only until this cartridge fills and I have to put in another
(approximately 1 week). I can make these files available in the future upon
request.

You can point your browser at ftp://whitewater.uoregon.edu/share/cameca/

or FTP anonymous to
whitewater.uoregon.edu

and look into the "/share/cameca/" directory

and find John's original Access file (xray.mdb, 704kb) ... you can also
find the Excel file I created with it, but it is 2Mb. I couldn't get Access
to save the file as XLS ... it created the file but the cells were empty
(?) ... I copied and pasted instead ... you may not be able to C&P if you
don't have enuf memory.
Let me know if you have any problems ...


TIA & cheerios, shAf
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/}
Michael Shaffer, R.A. - University of Oregon Electron Probe Facility
mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu
http://darkwing.uoregon.edu/~mshaf/epmahome/




From: HILDEGARD CROWLEY :      hcrowley-at-du.edu
Date: Tue, 22 Jul 1997 10:13:45 -0600 (MDT)
Subject: Re: Fix,wash,fix,store 5yrs

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Hi,

Long storage in glut is not good - the membranes are still permeable
since glut does not fix lipid. There will be an exodous of material and
changes. But - prefix, wash, postfix with osmium stabilizing the
membranes and the lipids, and store in buffer (not in alcohol - alcohol
removes osmium) for as long as needed. There is some movement of osmium,
but unless the storage is in a solvent, it is neglible.
Bye,
Hildy




From: Henry Bart :      bart-at-lasalle.edu
Date: Tue, 22 Jul 1997 13:00:49 -0400
Subject: SEM and Clay Mineral ID

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I need some help identifying clays using SEM. Do you
know of references, textbooks, etc. with pictures that would
me identify common clays i.e. illite, kaolinite, smectite,
chlorite, etc.

Hank Bart




From: sthomas-at-lanl.gov (Sharon C. Thomas)
Date: Tue, 22 Jul 1997 12:44:22 -0500
Subject: Help requested for maladjusted glass knifemaker

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The knifemaker available to me is a LKB Type 7801A from LKB Instruments Inc.
out of Rockville Maryland, but the company has since been taken over by
Leica/Leo. The knifemaker has two wheel type gauges that adjust the tension
from opposing sides on the glass piece. I can make glass squares, but when
I try to cut the square into the two knives, I don't get the correct
shapes, even
though I have tried systematically changing the settings on the two gauges
many different ways. Typically, the "sharp" edge may be chipped, the
reflection line in the glass is going in the opposite direction to what it
should
and the opposite edge to the sharp edge may be either sharp as well or too
thick a blunt edge. That's just one of the two knives; its' counterpart
often has
two "sharp" edges. Can anyone with any direct experience with this model of
knifemaker provide me with any advice on how to adjust the settings to produce
two glass knives?

Thanks in advance for any assistance that can be provided.






From: sthomas-at-lanl.gov (Sharon C. Thomas)
Date: Tue, 22 Jul 1997 12:44:22 -0500
Subject: Help requested for maladjusted glass knifemaker

Contents Retrieved from Microscopy Listserver Archives
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The knifemaker available to me is a LKB Type 7801A from LKB Instruments Inc.
out of Rockville Maryland, but the company has since been taken over by
Leica/Leo. The knifemaker has two wheel type gauges that adjust the tension
from opposing sides on the glass piece. I can make glass squares, but when
I try to cut the square into the two knives, I don't get the correct
shapes, even
though I have tried systematically changing the settings on the two gauges
many different ways. Typically, the "sharp" edge may be chipped, the
reflection line in the glass is going in the opposite direction to what it
should
and the opposite edge to the sharp edge may be either sharp as well or too
thick a blunt edge. That's just one of the two knives; its' counterpart
often has
two "sharp" edges. Can anyone with any direct experience with this model of
knifemaker provide me with any advice on how to adjust the settings to produce
two glass knives?

Thanks in advance for any assistance that can be provided.






From: MICHAEL DELANNOY :      delannoy-at-welchlink.welch.jhu.edu
Date: Tue, 22 Jul 1997 14:21:42 -0400 (EDT)
Subject: Uranyl formate

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People,
Does anyone know where to get uranyl formate? My previous
suppliers no longer make it-why? I do have the acetate, but the
formate works better with microfilaments. Any leads are welcome.

Searching in Baltimore,
Mike D.




From: Pat Hales :      hales-at-medcor.mcgill.ca
Date: Tue, 22 Jul 1997 14:47:11 -0700
Subject: EDTA Decalcification

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Last winter when I was making some changes to our tissue
processing/embedding protocol (which has been around here forever it seems),
I began asking questions about EDTA decalcification. One of our professors
told me that the EDTA decalcification was found to produce the fewest if any
artefacts IF (1) you use the disodium salt (not the tetrasodium), (2) it is
done at 4 C (not room temp), AND (3) decalcification is complete in 7-8
days. He claims that tissue pieces which take longer than that begin to
loose their membrane integrity. I have no references for this but he has
always been a reliable source of information in the past.

Pat Hales
McGill University
Dept. of Anatomy & Cell Biology
hales-at-hippo.medcor.mcgill.ca





From: wcornell-at-centum.utulsa.edu (Winton Cornell)
Date: Tue, 22 Jul 1997 13:40:25 -0500
Subject: EDS of zinc arsenides

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Micro-colleagues:

If some of you EDS practitioners (yes, I'm a WDS practitioner) could help me
with this I would be most appreciative:

1. presently I am involved in a project in which I have to examine zinc
arsenides - the process by which these are made can, at times, also produce
elemental Zn, ZnO and elemental As (yuk!, and then some)

2. I mainly characterize these beasties by x-ray diffraction, but in this
particular inspection I am doing some SEM work too

3. the samples I am looking at are suspected to be mainly the zinc arsenide
phase(s), however, I get EDS spectra with highly variable Zn/As - in fact,
more wide-ranging than expected for various of the possible zinc arsenides

4. I suspect that the Zn/As variation has as much to do with loss
(diminishment) of the As signal (due to absorption?) as it does with
variation in the ZnAs

5. to that end (#4), I performed a test yesterday in which I examined just
one crystal type (based on morphology) in areas of the sample where these
crystals were at the "surface" of the sample and at various "depths" within
holes and/or depressions.....basically, the deeper in the "hole" the lower
the As signal

6. concurrent with the dimished As, I also observed diminishment of Zn
L-alpha line (also a low-energy x-ray) such that Zn-K/Zn-L was also highly
variable - would you consider the latter a further indication of
absorption?

In the interim, as I await your response, I intend to run x-ray diffraction
on those samples with "apparently" low As (some are even As "absent") to
"confirm" whether or not the Zn or ZnO phases could be present. Together
(my XRD and preliminary SEM, plus your sage advice) I hope we can resolve this.

Thanks, in advance!!

Winton


P.S. if this message "surfaces" twice, please forgive me...the first time I
sent it I directed it to the listserver.....once discovered, some kind soul
might take it on himself/herself to forward it back to this list


Dr. Winton Cornell
Senior Research Associate & Supervisor, Microanalysis Laboratory
Department of Geosciences
The University of Tulsa
600 South College
Tulsa, OK 74104-3189

phone: 918-631-3248
fax: 918-631-2091
e-mail: wcornell-at-centum.utulsa.edu






From: Mike Coviello :      Coviello-at-mae.uta.edu
Date: Tue, 22 Jul 1997 15:24:11 -0500
Subject: Digital Darkrooms

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Hi All:
I recently put a message on the listserver requesting information on a
LogEtronics enlarger. Some responses I received suggested that I should
think about using a digital camera instead to capture images from negatives
and then process the images and then send them to a printer. I would like
some feedback from users. Is the quality close to that of film? Does anyone
have a system that they are extremely happy with? Are there any commercial
systems available? Please let me hear your experiences.

Thanks,

Michael Coviello
EM Lab Manager
The University of Texas -at- Arlington
Arlington, TX
E-mail coviello-at-mae.uta.edu
817-272-5496






From: Michael Shaffer :      mshaf-at-OREGON.UOREGON.EDU
Date: Tue, 22 Jul 1997 15:29:13 -0700
Subject: EPMA: new x-ray wavelength database

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Since my last post I have been working with the XLS spreadsheet file and
have added absorption edges, font highlighting, and Chuck's and John's
references. The new file name is xray_MS.XLS and is zipped as xray_MS.ZIP
...
Like I said before this will be a temporary FTP location for this file
... if Nestor is watching this thread, maybe he'll put the zipped file on
the MAS FTP site.
For those interested, I'm in the process of adding Cameca spectrometer
sine-theta values and creating a PDF file for the purpose of having this
data immediately available for on-line browsing ...

You can point your browser at ftp://whitewater.uoregon.edu/share/cameca/

or FTP anonymous to
whitewater.uoregon.edu

and look into the "/share/cameca/" directory

Let me know if you have any problems ... and PLEASE let me know if you
find any errors ...


TIA & cheerios, shAf
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/}
Michael Shaffer, R.A. - University of Oregon Electron Probe Facility
mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu
http://darkwing.uoregon.edu/~mshaf/epmahome/




From: ejb11-at-psu.edu (Edward J. Basgall)
Date: Tue, 22 Jul 1997 18:55:10 -0500
Subject: Re: Help requested for maladjusted glass knifemaker

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Sharon,
I would replace the cutting wheel first and then see what happens. The
systematic approach should work.

cheers

Edward J. Basgall, PhD
The Pennsylvania State University
Surface Chemistry Group ejb11-at-psu.edu
Materials Research Institute Building Ph: 814-865-0493
University Park, PA 16802-7003 FAX: 814-863-0618

{The knifemaker available to me is a LKB Type 7801A from LKB Instruments Inc.
{out of Rockville Maryland, but the company has since been taken over by
{Leica/Leo. The knifemaker has two wheel type gauges that adjust the tension
{from opposing sides on the glass piece. I can make glass squares, but when
{I try to cut the square into the two knives, I don't get the correct
{shapes, even
{though I have tried systematically changing the settings on the two gauges
{many different ways. Typically, the "sharp" edge may be chipped, the
{reflection line in the glass is going in the opposite direction to what it
{should
{and the opposite edge to the sharp edge may be either sharp as well or too
{thick a blunt edge. That's just one of the two knives; its' counterpart
{often has
{two "sharp" edges. Can anyone with any direct experience with this model of
{knifemaker provide me with any advice on how to adjust the settings to produce
{two glass knives?

{Thanks in advance for any assistance that can be provided.






From: A. Kent Christensen :      akc-at-umich.edu
Date: Tue, 22 Jul 1997 20:26:46 -0400 (EDT)
Subject: Re: Digital Darkrooms

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Mike

You will get lots of opinions on this. Here's mine. After spending
countless hours in the darkroom over the years, I've converted
enthusiastically to digital imaging, and now would prepare any serious
final labeled images with Photoshop, making the final prints on a
photographic quality printer. Does that mean that all original EM
pictures must be taken digitally? I don't think so. If you think about
it, only a small percentage of the EM images you take are apt to end up in
publications, projection slides or other serious uses. I find it much
easier to store numerous EM negatives as 3-1/4x4" sheets of film in
glassine envelopes than to fiddle with the multiple zip disks that are
necessary to store all the EM digital images in files large enough to
allow the resolution that may be necessary later on (the resolution level
that we take for granted in film negatives). So I would take them
initially on film (even though our department has a Philips EM100 fitted
with a CCD camera for digital imaging), and would observe the negatives on
a viewer (or on study prints) to decide what will be used for the final
product (publication figures, projection slides, etc.). For the final
pictures, I would generate digital images by scanning the chosen negatives
(with Leafscan 45 scanner) or scanning carefully-prepared prints (with a
Hewlett-Packard ScanJet II CX scanner), then crop, arrange, label and
otherwise fine tune the pictures with Photoshop 4 on a Power Mac 7600.
For photographic quality printing, we use a Kodak XLS 8600 PS printer.

For example, a very complex figure was prepared from multiple darkroom
prints, from which numerous small rectangles were cut out and mounted,
each showing polysomes at 100 kX mag. To convert to digital image, I
scanned the complex figure with the HP ScanJet, enlarging (exaggerating)
the image size so the file was 10-15 MB (to provide good resolution). The
scan was done without scanner sharpening (sharpening would be done in
Photoshop). The file was saved as a TIFF file, and taken up in Photoshop
4. Crop properly (rotate slightly, if necessary). The
brightness/contrast of the rectangles varied somewhat, so each was
selected (marquee), and Image} Adjust} AutoLevel was used to normalize (be
sure white=~5-7% and black=95% in Image} Adjust} Levels [or Curves]
eyedroppers). Sharpen whole figure with Filter} Sharpen} Unsharp Mask with
amount ~150-200%, radius 1, threshold 0. Dust or other small blemishes
(that are clearly of no scientific interest) can be removed with
Filter} Noise} Dust & Scratches (after minimal selection with marquee),
using radius 1-5 (no more than necessary to remove). Size (Image} Image
Size) the figure to width (or height) required by the journal, and set
resolution to 400-600 ppi. Put in figure number, size bar, and labels,
using layers (not directly on EM image). Print on Kodak XLS printer. The
image quality and detail compares well with work done in the darkroom.
More and more journals now allow you to submit such digital image files
for the final reproductions after a paper has been accepted.

Kent
(A. Kent Christensen, University of Michigan, akc-at-umich.edu)

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

On Tue, 22 Jul 1997, Mike Coviello wrote:

} I recently put a message on the listserver requesting information on a
} LogEtronics enlarger. Some responses I received suggested that I should
} think about using a digital camera instead to capture images from negatives
} and then process the images and then send them to a printer. I would like
} some feedback from users. Is the quality close to that of film? Does anyone
} have a system that they are extremely happy with? Are there any commercial
} systems available? Please let me hear your experiences.
}
} Michael Coviello
} EM Lab Manager
} The University of Texas -at- Arlington
} Arlington, TX
} E-mail coviello-at-mae.uta.edu
} 817-272-5496





From: Hall, Ernest L (CRD) :      hallel-at-exc01crdge.crd.ge.com (by way of
Date: Tue, 22 Jul 1997 19:49:54 -0500
Subject: Polymer Microscopy Position Avaialble

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Polymer Microscopy Position Available


The Microstructure and Microanalysis Program in the Characterization and
Environmental Laboratory at GE Corporate Research and Development,
Schenectady, NY, has an opening for a Polymer Microscopist at the Lead
Professional level. The primary duties associated with this position
involve the execution of research projects using Transmission Electron
Microscopy (TEM) to determine the structure of polymer blends and
coatings. Additional duties may involve research conducted using TEM
to study other materials, including metals, ceramics, or composites, or
using other microscopy techniques, including Acoustic Microscopy and
Atomic Force Microscopy.

The Characterization and Environmental Technology Laboratory is involved
in research into the structure and composition of materials in support
of development programs both at GE CRD and at GE businesses. Staff
members are expected to work independently with a high level of
expertise, and to become involved with a number of major project teams.
Good communication skills, both written and oral, are extremely
important.

The minimum requirements for this position are a MS in Materials Science
or a closely-related field and some prior experience with Transmission
Electron Microscopy. Demonstrated experience in polymer materials
science and characterization is also highly desirable.

Resumes and other information can be sent to:

Ernest L. Hall
Manager, Microstructure and Microanalysis Program
Room K1-2C12
GE Corporate Research and Development
PO Box 8
Schenectady, NY 12301
Fax: 518-387-6972
E-mail: hallel-at-crd.ge.com

g






From: Scott D. Walck WL/MLBT :      walcksd-at-ml.wpafb.af.mil
Date: Tue, 22 Jul 97 21:25:42 -0400
Subject: Re: Digital Darkrooms

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I'm going to a new position (PPG Industries) and I'm planning to implement a
digital darkroom instead of a conventional darkroom. The advantages are
tremendous.

At the Materials Directorate at Wright Lab, WPAFB, we have been using a Leaf
45 negative scanner. They are hard to find, but there is a company that has
updated it and is now selling it:

http://www.hyperzine.com/photokina/brem.html
http://www.hyperzine.com/photokina/brem2.html

I sent out a message several weeks ago on a web site that has information on a
collection of negative scanners, flatbed scanners, and some drum scanners:

http://www.foto.unibas.ch/scanners.html

Scitex used to make the Leaf, but now they have their flatbed smartscanners:

http://www.scitex.com/

Here's a site for information on doing it digitally:

http://www.presentingsolutions.com/adviceandinfodptoc.html

A site for drum scanners:
http://www.budde.com/products.htm
http://www.budde.com/print/magic.htm
http://www.budde.com/print/scanview.htm
http://www.budde.com/print/sm11000.htm

Our Leaf system is hooked into a MAC system with a lot of RAM, disk space, MO
drive, ZIP drive, and access to our LAN. This gives a lot of flexibilty with
different users. We bring in the images with a 16 bit format into
Photoshop, adjust the levels, convert the image to 8 bit grayscale with
appropriate gamma processing, invert the image into a positive print and save.
If the intermediate 4 x 5 setting of the Leaf 45 is used, a TEM negative can
be digitized with about 1200 dpi. If the image is printed to a 300 dpi
grayscale printer (i.e. sub-dye), then that gives an enlargement of 4x at the
printer. The Leaf system is capable of much higher pixel resolution and can
be used to digitize a very small area on the negative which can be printed
with the printer's 300 dpi setting providing a very high enlargement factor.
Photoshop has all the tools that could be done in the darkroom but are rather
tedious to do such as unsharp masking. Fixing scratches on negatives (mine
never have them), dodging, burning an other things that are required for
printing a good TEM negative can all be done in a few minutes. In addition,
it can be used on both MAC and PC platforms. John Russ's Image Processing
Toolkit provides plugins for Photoshop that can provide image processing and
stereology:
http://members.aol.com/imagproctk/index.htm

I put the scale markers into the print in layers, thus preserving the original
scanned image and saving in a TIF format provides the image in a compatible
format for other programs. I particularly like printing from Powerpoint onto
our Kodak 8650 printer. With Powerpoint, I copy several images of the Tif
file on one page (or several differnt files) and print. Output quality if
very good.

You need a fast computer with large RAM and large hard drives, a good size
monitor (minimum 17"), the scanner, pub-quality printer, and a portable-disk
large format disk drive. A 600 dpi laser printer is quite useful also. I
also use a flatbed scanner to digitize 6 TEM negatives in a Neg-a-file sheet
and digitize the page at 150 dpi. I also digitize my datasheet notes in
lineart format. These images are then put into my ThumbsPlus Image database
(shareware Program ~$50 for PC and Macs) I then have instant access to my
images. It sure beats making proof sheets and processing them.

All of this would probably be cheaper than going with a LogEtronics.

-Hopes this helps.

-Scott



} ------------------------------------------------------------------------ The
Microscopy ListServer -- Sponsor: The Microscopy Society of America To
Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
-----------------------------------------------------------------------.
}
} Hi All:
} I recently put a message on the listserver requesting information on a
LogEtronics enlarger. Some responses I received suggested that I should think
about using a digital camera instead to capture images from negatives and then
process the images and then send them to a printer. I would like some
feedback from users. Is the quality close to that of film? Does anyone have a
system that they are extremely happy with? Are there any commercial systems
available? Please let me hear your experiences.
}
} Thanks,
}
} Michael Coviello EM Lab Manager The University of Texas -at- Arlington
Arlington, TX E-mail coviello-at-mae.uta.edu 817-272-5496
}




From: Sarka Lhotak :      lhotaks-at-fhs.csu.mcmaster.ca
Date: Tue, 22 Jul 1997 22:24:06 -0400 (EDT)
Subject: immuno EM for actin; anti- BODIPY FL antibody

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Dear immuno-people,

I would like to summarize some of the answers I received concerning
the labelling of actin at EM level in non-muscle cells, namely chondrocytes
in the rabbit growth plate.

Rosemary White labelled actin in plants using monoclonal C4
anti-actin from ICN on LR White sections (Protoplasma 131:153-165 and
150:72-74). This antibody is mouse IgG against human actin and recognizes a
common actin epitope in many species.
Kirk Czymmek was successful using the antibody N.350 from Amersham
to label actin at the EM level in fungi (J Microscopy Vol 181, Feb 1996 pp
153-161; Protoplasma 163, pp. 199-202). This is a mouse IgM against chicken
gizzard actin and has been shown to label human, monkey, chicken, rat, etc
actin, therefore binding to a highly conserved region of actin.
BTW, we ordered this antibody today and hope to have some results
shortly!


Since we got nice labelling with phalloidin-BODIPY FL in the
confocal, we wanted to use rabbit anti-BODIPY FL antibody (Molecular
Probes) followed by anti-rabbit-gold to reveal the actin at EM level. This
was not successful. Molecular Probes could not give us any references of
anybody using this antibody for immunohistochemistry. Tamara Howard
commented, that though she has not used the anti-BODIPY, she had a dismal
luck with antibodies to FITC and Texas Red from various sources at the EM level.

Thanks to everybody who took the time and responded to my questions.
This list is such a wonderful way of sharing experience! I would never think
of looking in Protoplasma, since Medline does not bring it up!

Sarka Lhotak
EM Facility, McMaster University
Hamilton, Ontario, Canada






From: Mary Mager :      mager-at-unixg.ubc.ca
Date: Tue, 22 Jul 1997 22:38:19 -0700
Subject: Re: EDS of zinc arsenides

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Dear Winton,
If you look at particles in holes or depressions, the edges of the hole will
preferentially absorb the lower energy x-rays while the higher energy ones
can penetrate the edges of the hole. If you look at the As Ka line with the
SEM at 25 or 30 kV it will be less affected than the As L line. You will
also get better quantification. As with WDS, a flat, smooth sample would
also help.
You wrote:
} Micro-colleagues:
}
} If some of you EDS practitioners (yes, I'm a WDS practitioner) could help me
} with this I would be most appreciative:
}
} 1. presently I am involved in a project in which I have to examine zinc
} arsenides - the process by which these are made can, at times, also produce
} elemental Zn, ZnO and elemental As (yuk!, and then some)
}
} 2. I mainly characterize these beasties by x-ray diffraction, but in this
} particular inspection I am doing some SEM work too
}
} 3. the samples I am looking at are suspected to be mainly the zinc arsenide
} phase(s), however, I get EDS spectra with highly variable Zn/As - in fact,
} more wide-ranging than expected for various of the possible zinc arsenides
}
} 4. I suspect that the Zn/As variation has as much to do with loss
} (diminishment) of the As signal (due to absorption?) as it does with
} variation in the ZnAs
}
} 5. to that end (#4), I performed a test yesterday in which I examined just
} one crystal type (based on morphology) in areas of the sample where these
} crystals were at the "surface" of the sample and at various "depths" within
} holes and/or depressions.....basically, the deeper in the "hole" the lower
} the As signal
}
} 6. concurrent with the dimished As, I also observed diminishment of Zn
} L-alpha line (also a low-energy x-ray) such that Zn-K/Zn-L was also highly
} variable - would you consider the latter a further indication of
} absorption?
}
} In the interim, as I await your response, I intend to run x-ray diffraction
} on those samples with "apparently" low As (some are even As "absent") to
} "confirm" whether or not the Zn or ZnO phases could be present. Together
} (my XRD and preliminary SEM, plus your sage advice) I hope we can resolve this.
}
} Thanks, in advance!!
}
} Winton
}
}
} P.S. if this message "surfaces" twice, please forgive me...the first time I
} sent it I directed it to the listserver.....once discovered, some kind soul
} might take it on himself/herself to forward it back to this list
}
}
} Dr. Winton Cornell
} Senior Research Associate & Supervisor, Microanalysis Laboratory
} Department of Geosciences
} The University of Tulsa
} 600 South College
} Tulsa, OK 74104-3189
}
} phone: 918-631-3248
} fax: 918-631-2091
} e-mail: wcornell-at-centum.utulsa.edu
}
}
}
}
Mary Mager
Electron Microscopist
Metals and Materials Eng., UBC
6350 Stores Rd.
Vancouver, B.C. V6T 1Z4
CANADA
tel:604-822-5648, fax:604-822-3619
e-mail: mager-at-unixg.ubc.ca





From: Stephen A. Shaffer :      sshaffer-at-microdataware.com
Date: Tue, 22 Jul 1997 23:08:45 -0700
Subject: Inter/Micro 97 Day Two

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Continuing my reports on Inter/Micro 97: Today's sessions focused on
Instrumentation (AM) and Techniques (PM).

In the morning session, James M. Landrigan III of Polaroid discussed
their new Digital Microscope Camera ("DMC"). I have been anxious to see
what Polaroid has been up to and I am not surprised to report that their
new system will mark a significant addition to the development of
digital imaging. I won't risk mis-quoting the specs here since I didn't
yet get their literature, but I'll just mention that the system includes
a dedicated digital camera with a standard C-mount lens attachment which
should make it readily attachable to a range of microscopes (and other
equipment). The system also incorporates a larger (12.5mm?) pixel array
than competing systems which they claim results in an improved
signal-to-noise ratio. The camera includes more than 1,000,000 pixels
and produces images of either 800X600 or 1600X1200 pixels (other formats
too?) Part of the available resolution is from software interpolation,
which I'm not wild about, but I couldn't get clear just what the actual
pixel array was so you'll want to get more details from them. Also, one
factor which concerns me at first hearing is that they use a rectangular
pixel, not a square one. I'll want to ask them why that choice was
made. The price is about $6,000 for the camera and software system
which, while not cheap, certainly seems reasonable. All-in-all, I'm
happy to see Polaroid step into this field. Competition among the
giants can only help us users of the technology and this appears to be a
good entry into the fray.

Wayne Niemeyer of the McCrone Group showed some applications and results
of "Low Voltage Scanning Electron Microscopy." Those of you who know
the capabilities of these systems won't need "preaching to the choir"
but I must say that it really is impressive what can be done with these
systems. Wayne showed some beautiful images of exquisitely fine
structural features, some in complex, deeply three-dimensional surfaces,
all without any trace of charging, all in sharp, clear detail. I've
seen 'em before but I was impressed again.

John Reffner of Nicolet/Spectra-Tech reminded us that "There is
Microscopy in Infrared Microspectroscopy." He pointed out the
complimentary nature of microscopy and microspectroscopy, that a
scientist should be not a microscopist _or_ a spectroscopist, but both
if he or she wants to fully exploit the capabilities of these
instruments. It is appropriate that one from the company which actually
emphasizes the importance of good microscopy (and good microscopes) as a
part of microspectroscopy should make this point. Spectra-Tech is, in
my opinion, to microspectroscopy what Zeiss used to be to microscopy.
Yes, they may be the most expensive, but "you get what you pay for" here
as elsewhere. I think that we should advocate retaining the capability
to do good work with our equipment, even if we need to learn a bit more
to take advantage of that capability, rather than accept instruments
which have been "dumbed-down" to the least common denominator of the
people who use them. So far, Spectra-Tech has not succumbed to any
pressure they might feel to adopt that trend and I hope you'll join me
in encouraging them to continue to hold the high ground. Someone needs
to!

(Ok, ok, so I jamb in a bit of editorializing too. I'm not related to
any of the companies I'm mentioning, so I think I can use a bit of
license here.)

In the afternoon, Theodore M. Clarke of J.I. Case Corporation discussed
"High Magnification Photomacrography Using the Kodak 1.6i/AB MegaPlus
(TM) Camera." This provided a nice complement to the Polaroid talk, but
this by an independant technical evaluator who put the Kodak product
through some serious resolution tests, finding that it performed well if
you accepted Ted's recommended 500 X NA rule of thumb for maximum useful
magnification. This is 1/2 of the conventional wisdom but Ted made (and
illustrated) a good argument for the more conservative standard. The
Kodak product performed quite well in Ted's tests and I suggest all
interested parties watch for the publication of his work in The
Microscope in coming months. The technical detail of his work was too
much to repeat here, but watch for the publication as it will be well
worth using not only for its evaluation of the Kodak product but as a
model for how digital cameras can be well and truely tested.

Also in the afternoon, Allen Whiteside of the McCrone Group presented
two back-to-back papers on "Preparing Holey Carbon Films" and "AEM
Preparations Using Holey Carbon Film." (Note that "The McCrone Group"
is a different organization than the meeting hosts, the McCrone Research
Institute. They separated years ago.) Once again, the folks at the
McCrone Group demonstrate that they are masters of specimen manipulation
(i.e., particle handling) and specimen preparation as well as analytical
microscopy. I think they have probably originated more useful
techniques than any other single organization. Allen showed their
in-house technique for the preparation of holey carbon films which can
and should be tailored to the specific needs of the analysis at hand. A
film of one thickness, pore density, and web structure will be
appropriate for one analysis, something different for another. They
drop a solution of formvar in ethylene dichloride onto a glass slide
which is rotated on a turntable to spin out the applied fluid while the
solvent evaporates. The film is then lifted and applied to grids in a
more-or-less conventional manner but the secrets of the technique lie in
the speed of rotation and the formulation of the solvent/formvar
solution. Again, the idea is to experiment and determine the best film
preparation system for a particular kind of analysis, not adopt a single
procedure. There is more, or course, and this was the subject of
Allen's second talk. There are numerous techniques for applying the
particles of interest to the holey film, again chosen to best suit the
needs of the analysis.

I should close, but I just want to mention one more thing. There are
many more papers being presented than I'm mentioning here. I'm only
hitting on a few that seem particulary interesting to me for one odd
reason or another. One should not infer anything from my failing to
mention some of the other fine papers!

More to come, stay tuned!

Steve Shaffer
MicroDataware

p.s. Yes, we are till publishing the Particle Atlas Electronic Edition
(PAEE). Please contact me via EMail if you would like further
information.

p.p.s. I know I'm a terrible speller and I'm preparing these things
quickly while on the road and without access to my regular array of
make-me-look-good software, so please forgive the rough nature of these
communications. ;-)




From: Benchaib Mehdi :      benchaib-at-rockefeller.univ-lyon1.fr
Date: Wed, 23 Jul 1997 10:02:13 +0200 (MET DST)
Subject: unsubscribe

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Dr Mehdi Benchaib
Laboratoire d' Histologie, Embryologie et Biologie de la Reproduction
8 avenue Rockefeller F-69373 Lyon CEDEX 08






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Dr Mehdi Benchaib
Laboratoire d' Histologie, Embryologie et Biologie de la Reproduction
8 avenue Rockefeller F-69373 Lyon CEDEX 08






From: GREG VAUGHN MARTIN :      gmartin-at-welchlink.welch.jhu.edu
Date: Wed, 23 Jul 1997 08:29:05 -0400 (EDT)
Subject: Re: Digital Darkrooms

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Hey Mike and everyone --

I've dealt with both 35mm (immunofluorescence) and 3 1/4 X 4 1/4 inch
(EM) negatives in both ways. Using a high quality light box and a very
high quality macro lens on a fairly standard sort of CCD camera (Dage 72,
640X480, 8bits). I've gotten good digital images, printed on a Codonics
dye-sub. But (my humble opinion) the quality doesn't approach what you can do
in the darkroom. It is, however, WAY faster and easier. A fancier
camera may help, but it all depends on what you want to do with the
images. If the images are already on film, then I would tend to stick
with silver grains over pixels.
If you really need to go digital and have some money, scanners are
probably the way to go for digitizing negatives -- check the list's archives,
there has been much discussion of scanners.

Greg Martin
Dept. of Cell Biology and Anatomy
Johns Hopkins School of Medicine

On Tue, 22 Jul 1997, Mike Coviello wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Hi All:
} I recently put a message on the listserver requesting information on a
} LogEtronics enlarger. Some responses I received suggested that I should
} think about using a digital camera instead to capture images from negatives
} and then process the images and then send them to a printer. I would like
} some feedback from users. Is the quality close to that of film? Does anyone
} have a system that they are extremely happy with? Are there any commercial
} systems available? Please let me hear your experiences.
}
} Thanks,
}
} Michael Coviello
} EM Lab Manager
} The University of Texas -at- Arlington
} Arlington, TX
} E-mail coviello-at-mae.uta.edu
} 817-272-5496
}
}
}




From: George.C.Ruben-at-Dartmouth.EDU (George C. Ruben)
Date: 23 Jul 97 09:01:32 EDT
Subject: normal versus digital darkroom

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

Mike,

I would buy a Durst 1200 point source
enlarger if you are doing high resolution electron microscopy ---- the
logEtronics enlarger uses a scanning spot to equalize large contrast variations
within an image and this enlarger prints like a diffuse source enlarger not a
point source enlarger. We have found that lines are sharper and narrower with
the point source enlarger.
Our work requires darkroom printing of reversal negatives made
from thin vertically Pt-C replicated specimens in which we want to visualize
molecular details on the structural highlights. Usually these features only
become visible after printing the reversal negative on fiber based paper and
drying the print on glossy paper----
For TEM publications that do not need this detail as part of
their story, it is easy to scan in images at a hardware resolution of 600 dpi
and label the image with Adobe photoshop---- I find that I need access to a
good darkroom as well as the appropriate digital darkroom equipment.

George Ruben
Dept. Biological Sciences
Dartmouth College
Hanover, NH 03755




From: C. John Runions :      cjr14-at-cornell.edu
Date: Wed, 23 Jul 1997 10:21:47 -0400
Subject: GMA polymerization

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X-Sender: cjr14-at-postoffice.mail.cornell.edu
Message-Id: {v03020900affbc2e66cb8-at-[132.236.111.203]}
Mime-Version: 1.0
Content-Type: text/plain; charset="us-ascii"

Hi everyone, for many years I have been embedding in Spurr's resin but
have recently decided to give glycol methacrylate a whirl for thicker LM
sections of plant material. Many years have passed since my last work with
GMA. I remember that we made a plexigalss container which we could
evacuate the oxygen from by flushing with nitrogen and that polymerization
was then carried out below a longwave UV light placed in the container.
Does this sound correct? If so does anyone have any plans or tricks that
might assist me in constructing such a device? Cheers, John

=================
C. John Runions, Ph.D
Section of Ecology and Systematics
Corson Hall
Cornell University
Ithaca, New York
USA 14853

email cjr14-at-cornell.edu [ie. cjr(fourteen)-at-...]
phone (607) 254-4282
Fax (607) 255-8088






From: Walt Bobrowski (313) 996-7814 :      bobroww-at-aa.wl.com
Date: Wed, 23 Jul 1997 10:56:01 -0400 (EDT)
Subject: RE: Digital Darkroom

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We've gone digital TEM and everyone loves it for ease of capture and speediness
to the investigator. Our 2Kx2K images look great on a large monitor and
printed with on a Tektronix dye-sub printer. Heck, even an Epson Stylus
printer with 1440 dpi makes darn good prints (on photo-quality paper)! In a
side-by-side comparison, I'll take a high contrast print from our Durst EM1200
over the dye-sub. However, dollar for dollar, one alternative to consider is to
transfer TEM negatives to Kodak PhotoCD and print only select images, when you
need publication quality, to the Fujix Pictrograph 3000 digital printer (which
costs less than the Durst with all the electronic bells and whistles.) Thereby
you have your negative, a high-resolution digital image, a digital print, and
you can still make a print if you really have to.

There's my humble opinion. Thank you.

Walter F. Bobrowski
Subcellular Pathology
Parke-Davis Pharmaceutical Research
Ann Arbor, MI 48105

TEL: 313-996-7814
FAX: 313-996-5001
E-Mail: BOBROWW-at-AA.WL.COM





From: Doug Cromey :      doug-cromey-at-ns.arizona.edu
Date: Wed, 23 Jul 1997 08:02:14 -0700
Subject: Re: normal versus digital darkroom

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To all who would advocate a totally digital darkroom,

While on the whole I have to agree that the direction that
science/microscopy is taking is a digital one, I must point out that there
are some serious shortcomings to digital imaging. The most serious, in my
mind, is the issue of archivability. While I could launch into this
myself, I'd prefer to bring to everyone's attention the following article
(which addresses this issue far better than I can):

"Ensuring the Longevity of Digital Documents"
Jeff Rothenberg, Scientific American, January 1995

If we can't honestly answer the question of how we, or anyone else, will be
able to use (or even access) our digital images 30 years from now, we need
to re-evaluate the speed with which we are going digital. If properly
cared for, film can last for decades. The uncertain aging of digital
storage media, the often rapid obsolescence of drive mechanisms & media
types and the ongoing changes in image file formats are cause for concern.

Yours,
Doug Cromey
.....................................................................
: Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy :
: Sr. Research Specialist 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




From: Tom Phillips :      tphillips-at-biosci.mbp.missouri.edu
Date: Wed, 23 Jul 1997 10:32:48 -0500
Subject: CD-ROM's for archiving - any experience?

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Our multi-user facility is currently archiving our confocal and LM digital
images on Panasonic optical disks (re-writable, very stable -at- about $125
for 1 GB). The disadvantage is that few of our users have their own
Panasonic drives so most people simply archive the images at our core and
then move the ones they want by FTP as needed. I would like to switch to a
more universal medium - namely CD ROM's. My understanding is that CD's can
now be written to in multiple sessions so you don't need to fill an entire
disk at once. Furthermore, it is my understanding that a disk of TIFF
images should be readable by both IBM/WINTEL and Mac/PowerPC types
computers. Is anybody actually doing this? Comments on how reliable are
the recorders, which ones are best, pitfalls, etc would be appreciated.
Before I get a dozen advocates of ZIP/Jazz drives, I don't want to go that
route since that they are not as ubiquitous as CD drives. Thanks in
advance.


Thomas E. Phillips, Ph.D.
Associate Professor of Biological Sciences
Director, Molecular Cytology Core Facility
3 Tucker Hall
University of Missouri
Columbia, MO 65211
(573)-882-4712 (voice)
(573)-882-0123 (fax)






From: joyce craig :      bafpjec-at-csu.edu
Date: Wed, 23 Jul 1997 23:40:29 -0700
Subject: maladjusted glass knifemaker

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The only time I have had problems similar to what you describe it was
when I had a bad cutting wheel. Have you changed it recently?




From: Barr, Dennis :      dennbarr-at-eastman.com
Date: Wed, 23 Jul 1997 13:33:48 -0400
Subject: RE: Digital Darkrooms

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Mike, your needs for image quality may exceed ours (industrial research
laboratory, not as interested in publishing outside as academic), but we
recently removed all of our darkrooms because we had not processed a
negative or print in two years.

All of our microscopy (AFM, TEM, SEM, and Optical) is done with digital
image capture. The speed to get a hardcopy, the flexibility of sharing
images worldwide via electronic transfer, and the cost per print/image
all strongly favor total digital imaging. Although the quality of a
typical digital image does not equal that which can be obtained with
photographic film they meet our needs completely. For the highest
quality digital imaging in optical microscopy we use a Kodak 460 digital
camera or a Leaf MicroLumina (3500 X 2400 pixels). Storage of images is
done on CD-ROM for archival capability and erasable Magneto-Optical CD
(40GB jukebox on a network server) is used for day-to-day image storage.
Photoshop is used for image "retouching". Reports are composed in
MicroSoft Word and hardcopy of images are obtained with networked Kodak
8650 printers (one dedicated to B/W and one for color).

As a final note, we've been using video and digital images for 9 years
and would never think of going back to a photographic imaging process.

*********************************************************
Dr. Dennis B. Barr
Research Laboratories
Eastman Chemical Company
Kingsport, TN 37664-5150
Phone:423-229-2188
Fax:423-229-4558
Email: dennbarr-at-eastman.com
********************************************************

} -----Original Message-----
} From: Mike Coviello [SMTP:Coviello-at-mae.uta.edu]
} Sent: Tuesday, July 22, 1997 4:24 PM
} To: Microscopy-at-Sparc5.Microscopy.Com
} Subject: Digital Darkrooms
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America




From: Larry Ackerman :      mishot-at-itsa.ucsf.edu
Date: Wed, 23 Jul 1997 11:02:53 -0700
Subject: Digital Darkrooms

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If you spend $30,000--$50,000 for hardware and software (which will be
obsolete in less than five years) you can produce very good hardcopy images
comparable to conventional photography. The digital process requires a
highly skilled system caretaker and the learning period for neophytes is
much longer. For every year that you can delay the transition to digital you
will reduce the problems significantly. These statements are based on my
experiences with photography for 32 years and digital graphics for 10 years.
Larry D. Ackerman (415) 476-8751
Howard Hughes Medical Institute FAX (415) 476-5774
UCSF, Box 0724, Rm U426
533 Parnassus Ave. mishot-at-itsa.ucsf.edu
San Francisco, CA 94143





From: Crossman, Harold :      crossman-at-OSI.SYLVANIA.com
Date: Wed, 23 Jul 1997 14:46:32 -0400
Subject: RE: Digital Darkrooms

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On the other hand...
I can send digital images to any of my JIT-based manufacturing customers
in a matter of moments via e-mail. Do that with film!
I can store images with and without annotation in an inexpensive
database that is continually being upgraded by its developers... and is
forward and backward compatible with our standard desktop applications.
I have reduced my Polaroid film expenditures by at least five thousand
dollars per year (with the attendant costs to order, deliver, bill,
etc.)
My image acquisition system is never out of stock.
Images don't get "lost in the mail"
I can extract a variety of numbers from a digital image easier than
using primitive tools like an ASTM grain size overlay.

It should be noted that the medical industry is a driving force for
improvements in digital imaging technologies and electronic
collaboration precisely because of the failure of film to fulfill the
needs of today's customers.



} ------------------------------------------------
} Opinions or statements expressed herein, rational or otherwise, do not
} necessarily reflect those of my employer.
}
} Harold J. Crossman
} OSRAM SYLVANIA INC.
} Lighting Research Center
} 71 Cherry Hill Dr.
} Beverly, MA 01915
} Phone: (508) 750-1717
} E-mail: crossman-at-osi.sylvania.com
}
} Our web sites: www.sylvania.com
} www.siemens.com
} --
}
} "Crossman, Harold" {crossman-at-osi.SYLVANIA.com}




From: John Turek :      jjt-at-vet.purdue.edu
Date: Wed, 23 Jul 1997 14:36:55 -0500
Subject: CD-ROM's for archiving - any experience?

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Tom:

We use CD-ROMs for archiving images, data, etc. If you make the CD-ROM in
ISO-9660 format it can be read by PC's, Macs, and Unix machines. ISO-9660
does not allow long file names. There are other formats - Joliet system -
that allow long file names but these can only be read in Win95 machines. I
recommend that you have a dedicated machine for making CD-ROMS. Partition
the hard drive so that your system files are on C-drive and leave the
D-drive for files to be archived. Two of the major manufacturs are Yamaha
and Pinnacle Micro. If you look up their web-sites and read the FAQ's
related to installation and troubleshooting, you will get some idea of the
important issues in setting up a system (There are certain hard drive
specifications, etc.) Multisession is possible, but you need software that
will read a multisession disk (usually the software package that was used
to make the CD). If you make a multisession disk, and place it in a
computer without the proper software - the computer will only see the last
session. This drawback may be changing (changed?) with the next generation
of machines and software. Overall, I think CD-ROM is the current best
method for archiving data.

Regards,

John J. Turek, Ph.D.
Associate Professor
Director, Electron Microscopy Laboratory and Core
Laboratory for Image Analysis and Multidimensional
Applications (CRISTAL)
Department of Basic Medical Sciences
1246 Lynn Hall, G193C
Purdue University
W. Lafayette, IN 47907-1246
Phone: 765-494-5854
Fax: 765-494-0781
Email: jjt-at-vet.purdue.edu







From: Post Doc :      sinkler-at-apollo.numis.nwu.edu
Date: Wed, 23 Jul 1997 15:19:13 -0500 (CDT )
Subject: Re: CD-ROM's for archiving - any experience?

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


Tom:

We have installed writable CD-ROM on a UNIX system using software
manufactured by Microson, called GEAR 32.

There have been serious problems arising from incompatibility of UNIX
with CD-ROM technology (asynchronous versus synchronous - The Unix
machines deliver information when they are ready but the CD writing
process requires information at a constant rate which is determined by
the disc speed). I don't believe this is a problem with PC's, but watch
out if you want to use CD-R on a UNIX platform. We had to buy a new
external hard drive, directly connected to the CD-R device, in order to
get everything to work reliably.

Also, and this may apply to you as well, the claims of the software
manual concerning various options like multisession backing up were not
actually implementable. We must write an entire CD at once. This is is
not so bad actually as we deal with large quantities of image data, and
the disks themselves are now only around $4-$5, which you really can't
beat for 650MB of space. The software packages available when working
in the PC world may be better, but beware, and make sure you get what
is advertised. I think this technology has a future and that the
microscopy world can benifit. However, the software for running it, at
least on a UNIX platform, has some progress still ahead of it.

Wharton

++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Wharton Sinkler PhD
Department of Materials Science and Engineering
Northwestern University
2225 North Campus Drive
Evanston, IL 60208-3108
tel: (847) 491-7809
fax: (847) 491-7820
email: sinkler-at-apollo.numis.nwu.edu

On Wed, 23 Jul 1997, Tom Phillips wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Our multi-user facility is currently archiving our confocal and LM digital
} images on Panasonic optical disks (re-writable, very stable -at- about $125
} for 1 GB). The disadvantage is that few of our users have their own
} Panasonic drives so most people simply archive the images at our core and
} then move the ones they want by FTP as needed. I would like to switch to a
} more universal medium - namely CD ROM's. My understanding is that CD's can
} now be written to in multiple sessions so you don't need to fill an entire
} disk at once. Furthermore, it is my understanding that a disk of TIFF
} images should be readable by both IBM/WINTEL and Mac/PowerPC types
} computers. Is anybody actually doing this? Comments on how reliable are
} the recorders, which ones are best, pitfalls, etc would be appreciated.
} Before I get a dozen advocates of ZIP/Jazz drives, I don't want to go that
} route since that they are not as ubiquitous as CD drives. Thanks in
} advance.
}
}
} Thomas E. Phillips, Ph.D.
} Associate Professor of Biological Sciences
} Director, Molecular Cytology Core Facility
} 3 Tucker Hall
} University of Missouri
} Columbia, MO 65211
} (573)-882-4712 (voice)
} (573)-882-0123 (fax)
}
}
}






From: Post Doc :      sinkler-at-apollo.numis.nwu.edu
Date: Wed, 23 Jul 1997 15:19:13 -0500 (CDT )
Subject: Re: CD-ROM's for archiving - any experience?

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


Tom:

We have installed writable CD-ROM on a UNIX system using software
manufactured by Microson, called GEAR 32.

There have been serious problems arising from incompatibility of UNIX
with CD-ROM technology (asynchronous versus synchronous - The Unix
machines deliver information when they are ready but the CD writing
process requires information at a constant rate which is determined by
the disc speed). I don't believe this is a problem with PC's, but watch
out if you want to use CD-R on a UNIX platform. We had to buy a new
external hard drive, directly connected to the CD-R device, in order to
get everything to work reliably.

Also, and this may apply to you as well, the claims of the software
manual concerning various options like multisession backing up were not
actually implementable. We must write an entire CD at once. This is is
not so bad actually as we deal with large quantities of image data, and
the disks themselves are now only around $4-$5, which you really can't
beat for 650MB of space. The software packages available when working
in the PC world may be better, but beware, and make sure you get what
is advertised. I think this technology has a future and that the
microscopy world can benifit. However, the software for running it, at
least on a UNIX platform, has some progress still ahead of it.

Wharton

++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Wharton Sinkler PhD
Department of Materials Science and Engineering
Northwestern University
2225 North Campus Drive
Evanston, IL 60208-3108
tel: (847) 491-7809
fax: (847) 491-7820
email: sinkler-at-apollo.numis.nwu.edu

On Wed, 23 Jul 1997, Tom Phillips wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Our multi-user facility is currently archiving our confocal and LM digital
} images on Panasonic optical disks (re-writable, very stable -at- about $125
} for 1 GB). The disadvantage is that few of our users have their own
} Panasonic drives so most people simply archive the images at our core and
} then move the ones they want by FTP as needed. I would like to switch to a
} more universal medium - namely CD ROM's. My understanding is that CD's can
} now be written to in multiple sessions so you don't need to fill an entire
} disk at once. Furthermore, it is my understanding that a disk of TIFF
} images should be readable by both IBM/WINTEL and Mac/PowerPC types
} computers. Is anybody actually doing this? Comments on how reliable are
} the recorders, which ones are best, pitfalls, etc would be appreciated.
} Before I get a dozen advocates of ZIP/Jazz drives, I don't want to go that
} route since that they are not as ubiquitous as CD drives. Thanks in
} advance.
}
}
} Thomas E. Phillips, Ph.D.
} Associate Professor of Biological Sciences
} Director, Molecular Cytology Core Facility
} 3 Tucker Hall
} University of Missouri
} Columbia, MO 65211
} (573)-882-4712 (voice)
} (573)-882-0123 (fax)
}
}
}






From: shAf :      mshaf-at-darkwing.uoregon.edu
Date: Wed, 23 Jul 1997 13:11:46 -0700
Subject: Re: archiving digital

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

Doug Cromey wrote:

} ...
}
} . . . The most serious, in my mind, is the issue of archivability.
} ...
}
} "Ensuring the Longevity of Digital Documents"
} Jeff Rothenberg, Scientific American, January 1995
}
} If we can't honestly answer the question of how we, or anyone else,
} will be
} able to use (or even access) our digital images 30 years from now, we
} need
} to re-evaluate the speed with which we are going digital. If properly
}
} cared for, film can last for decades. The uncertain aging of digital
} storage media, the often rapid obsolescence of drive mechanisms &
} media
} types and the ongoing changes in image file formats are cause for
} concern.
}
} ...

I agree ... however, while there exists good reasoning to avoid
magnetic media for long-term archiving, there seems to be little
concern in this regard for magneto-optical or CD ROM media. Also
regarding archival quality ... digital hardcopy will not hold up to
long-term storage of preperly washed photographic papers (... I think it
has already been mentioned that the resolution and the quality of the
grayscale for even the best digital printers doesn't even come close to
chemica darkroom printing ...).
Still ... even as an ex-photographic art student ... I can appreciate
the new capabilities (in general) that a digital darkroom has over the
traditional for producing *lots of work* and its being "publication
quality" ... if not "art for the purist" quality ...


cheerios, shAf
--
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/}
Michael Shaffer, R.A. - University of Oregon Electron Probe Facility
mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu
http://darkwing.uoregon.edu/~mshaf/






From: Scott Whittaker :      sdw-at-biotech.ufl.edu
Date: Wed, 23 Jul 1997 16:39:37 -0400
Subject: Re: CD-ROM's for archiving - any experience?

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

Hi Tom. I have been burning CD's for our SEM users for about a year and a
half now. We have an HP 4020i and I am using the Easy CD software package
(supposedly the best).
Of the 50 or so users who have archived onto CD only 2 have not been able
to get images I wrote and another says he can't but I think in his case it
is the operator. Both of the users who are having trouble are able to read
the data from the first write, but not from successive write sessions.
Thay have brought their disks back into the lab and I am able to read ALL
the data from ALL write sessions on each of our four cd readers. Our
computer guy here asked how old the drives they were using to read the
disks were cause if they are too old they may not support the multi-session
standards. Only one user seems to have such an aged cd rom. The other is
baffeling. Heck, my cd-rom at the house is old as the hills (and the
cheapest I could find) and it does fine.
HP was no help in this matter either so if anyone has any ideas I would
sure appreciate hearing from you. I can still say I reccommend the writer
and feel it is probably the best deal. I don't feel bad making the users
buy an $8-9 disk, even the guys who can only read the first session. You
are correct that both MAC & PC's will read the files as long as the MAC is
running system 7 or better. SGI will not nor will OS2. Good luck





At 10:32 AM 7/23/97 -0500, you wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America




} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {
GO GATORS
Scott D. Whittaker 218 Carr Hall
Research Assistant Gainesville, FL 32610
University Of Florida ph 352-392-1295
ICBR EM Core Lab fax 352-846-0251
sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/
The home of " Tips & Tricks "









From: Robert Underwood :      underwoo-at-u.washington.edu
Date: Wed, 23 Jul 1997 14:24:22 -0700 (PDT)
Subject: Re: CD-ROM's for archiving - any experience?

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

Hi Tom,

We have been archiving to CDs with a Philips 2000 for almost two years
with no problems. Much cheaper than opticals. $9.00 for 600 MB.

Bob
Morphology Core
Seattle

On Wed, 23 Jul 1997, Tom Phillips wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Our multi-user facility is currently archiving our confocal and LM digital
} images on Panasonic optical disks (re-writable, very stable -at- about $125
} for 1 GB). The disadvantage is that few of our users have their own
} Panasonic drives so most people simply archive the images at our core and
} then move the ones they want by FTP as needed. I would like to switch to a
} more universal medium - namely CD ROM's. My understanding is that CD's can
} now be written to in multiple sessions so you don't need to fill an entire
} disk at once. Furthermore, it is my understanding that a disk of TIFF
} images should be readable by both IBM/WINTEL and Mac/PowerPC types
} computers. Is anybody actually doing this? Comments on how reliable are
} the recorders, which ones are best, pitfalls, etc would be appreciated.
} Before I get a dozen advocates of ZIP/Jazz drives, I don't want to go that
} route since that they are not as ubiquitous as CD drives. Thanks in
} advance.
}
}
} Thomas E. Phillips, Ph.D.
} Associate Professor of Biological Sciences
} Director, Molecular Cytology Core Facility
} 3 Tucker Hall
} University of Missouri
} Columbia, MO 65211
} (573)-882-4712 (voice)
} (573)-882-0123 (fax)
}
}
}





From: shAf :      mshaf-at-darkwing.uoregon.edu
Date: Wed, 23 Jul 1997 16:18:50 -0700
Subject: Re: CD-ROM's for archiving - any experience?

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









Pat Sileo
07/11/97 10:47 AM

To: Bob Holthausen/SLSNY/Pall/US-at-Pall
cc:

} ...
} }
} } Our multi-user facility is currently archiving our confocal and LM
} digital
} } images on Panasonic optical disks (re-writable, very stable -at- about
} $125
} } for 1 GB). The disadvantage is that few of our users have their own
}
} } Panasonic drives so most people simply archive the images at our
} core and
} } then move the ones they want by FTP as needed. I would like to
} switch to a
} } more universal medium - namely CD ROM's. My understanding is that
} CD's can
} } now be written to in multiple sessions so you don't need to fill an
} entire
} } disk at once. Furthermore, it is my understanding that a disk of
} TIFF
} } images should be readable by both IBM/WINTEL and Mac/PowerPC types
} } computers. Is anybody actually doing this? Comments on how
} reliable are
} } the recorders, which ones are best, pitfalls, etc would be
} appreciated.
} } Before I get a dozen advocates of ZIP/Jazz drives, I don't want to
} go that
} } route since that they are not as ubiquitous as CD drives. Thanks
} in
} } advance. ...

Whereas I would have thought CDROM should be the best method (...
considering the cost of the media ...), judging from the responses we've
seen I'm glad I went with a Fujitsu 640 magneto-optical. It has a solid
SCSI interface (... I use Adaptec ...) and it is re-writable. I had
some early issues when it first came out (... we had been using 230Mb
for 1.5 years ...) but it works flawlessly now. The cost per 640Mb
cartridge is (however) $35 ... and (... of course ...) the drives aren't
as ubiquitous. We use it in a intranet configuration, and all image
files and data from the SEM and Cameca archive directly to it.
Non-WIntel users download from it via "anonymous" FTP ...

... another point of view ...


cheerios, shAf
--
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/}
Michael Shaffer, R.A. - University of Oregon Electron Probe Facility
mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu
http://darkwing.uoregon.edu/~mshaf/






From: wcornell-at-centum.utulsa.edu (Winton Cornell) (by way of Nestor J.
Date: Wed, 23 Jul 1997 18:52:19 -0500
Subject: EDS of zinc arsenides

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

Micro-colleagues:

If some of you EDS practioners (yes, I'm a WDS practioner) could help me
with this I would be most appreciative:

1. presently I am involved in a project for which I have to examine zinc
arsenides - the process by which these are made can, at times, also produce
elemental Zn, ZnO and elemental As (yuk!, and then some)

2. I mainly characterize these beasties by x-ray diffraction, but in this
particular inspection I am doing some SEM work too

3. the samples I am looking at are suspected to be mainly the zinc arsenide
phase(s), however, I get EDS spectra with highly variable Zn/As - in fact,
more wide-ranging than expected for various of the possible zinc arsenides

4. I suspect that the Zn/As variation has as much to do with loss
(diminishment) of the As signal (due to absorption?) as it does with
variation in the ZnAs

5. to that end (#4), I performed a test yesterday in which I examined just
one crystal type (based on morphology) in areas of the sample where these
crystals were at the "surface" of the sample and at various "depths" within
holes and/or depressions.....basically, the deeper in the "hole" the lower
the As signal

6. concurrent with the dimished As, I also observed diminishment of Zn
L-alpha line (also a low-energy x-ray) such that Zn-K/Zn-L was also highly
variable - would you consider the latter a further indication of
absorption?

In the interim as, I await your response, I intend to run x-ray diffraction
on the samples with "apparently" low As (some are even As "absent") to
"confirm" whether or not the Zn or ZnO phases could be present. Together
(my XRD and preliminary SEM, plus your sage advice) I hope to resolve this.

Thanks, in advance!!

Winton


Dr. Winton Cornell
Senior Research Associate & Supervisor, Microanalysis Laboratory
Department of Geosciences
The University of Tulsa
600 South College
Tulsa, OK 74104-3189

phone: 918-631-3248
fax: 918-631-2091
e-mail: wcornell-at-centum.utulsa.edu






From: Jim Darley :      jim-at-proscitech.com.au
Date: Thu, 24 Jul 1997 10:22:24 +1000
Subject: Re: Help requested for maladjusted glass knife maker

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

Sharon et.al.
That knife maker is an everlasting instrument but the adjustments must be
right, otherwise its a big waste of glass and time.
1 Adjust the back holder of the glass, so that the cutting stroke across
the diagonal of the glass square stops one or two mm before running off the
glass.
2 Loosen the front holder and press the moving part to just touch the
corner of the glass. Increase the push against the spring tension by two
scale divisions and then tighten the knurled knob.
3 The front and back lateral adjustments must now be set. Centre both of
these, check with a glass square inserted and a piece of paper as a
straight edge that the centre line would be corner to corner on the glass.
4 Now adjust the back lateral adjustment so that the centre line would be
1 to 2mm to the left of the back corner. Tighten the lock-nut.
5 Repeat for the front adjustment but to the right of the front corner.
6 Break a couple of test squares, without the front damper, which pushes a
bit of rubber against the glass, applied. Those knives should have their
edges close, but not across the corners. If required adjust the lateral
controls.
7 Once well set, the laterals never need adjustment. Draw a marker pen
line across the dials to indicate settings or tighten them just beyond
finger strengths.

Never touch the clamping head when it is lowered; it determines the
clamping pressure and over-tightening its locking device is pointless.

The rubber damping device slows down the moment of fracture of the front
knife only. This result in a wider stress free area for that knife.
Apply the damper after clamping the head.
Mover rubber front to just touch glass plus two scale divisions. The glass
should not move back. Score, break and then, always before lifting the
clamping head, release the damper.

I am writing this from memory but I've taught that operation a few hundred
times.
Ask me if you still have problems with the knife maker.
Disclaimer: ProSciTech and most other EM suppliers supply microtomy glass.
We should have an interest in maladjusted knife-makers.
Cheers
Jim Darley

ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313
Great microscopy catalogue, 400+ Links, MSDS
************************ http://www.proscitech.com.au

----------
} From: Sharon C. Thomas {sthomas-at-lanl.gov}
} To: Microscopy-at-sparc5.microscopy.com
} Subject: Help requested for maladjusted glass knifemaker
} Date: Wednesday, 23 July 1997 3:44
} The knifemaker available to me is a LKB Type 7801A from LKB Instruments
Inc.
} out of Rockville Maryland, but the company has since been taken over by
} Leica/Leo. The knifemaker has two wheel type gauges that adjust the
tension
} from opposing sides on the glass piece. I can make glass squares, but
when
} I try to cut the square into the two knives, I don't get the correct
} shapes, even
} though I have tried systematically changing the settings on the two
gauges
} many different ways. Typically, the "sharp" edge may be chipped, the
} reflection line in the glass is going in the opposite direction to what
it
} should
} and the opposite edge to the sharp edge may be either sharp as well or
too
} thick a blunt edge. That's just one of the two knives; its' counterpart
} often has
} two "sharp" edges. Can anyone with any direct experience with this model
of
} knifemaker provide me with any advice on how to adjust the settings to
produce
} two glass knives?
}
} Thanks in advance for any assistance that can be provided.
}
}




From: Larry Allard :      allardlfjr-at-ornl.gov
Date: Wed, 23 Jul 1997 22:59:56 -0400
Subject: Re: CD-ROM's for archiving - any experience?

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


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

Tom:

While we have do many Zip drives in use in our multi-user facility, they
are used for purposes other than archive storage. We archive all of our
digital images (from 6 major instruments) onto CD-ROMS. We have more than
60 Gbytes of active, primary storage on on-line servers, which is good
enough for 3-4 months of image storage, after which the oldest images are
downloaded onto CDs. This has worked nicely for us for the last 3-4 years.
We store all our images in their original formats (DigitalMicrograph, Adobe
PhotoShop etc.) so they can be handled ultimately just as if they had just
been recorded.

Hope this helps.

Larry



}
} Our multi-user facility is currently archiving our confocal and LM digital
} images on Panasonic optical disks (re-writable, very stable -at- about $125
} for 1 GB). The disadvantage is that few of our users have their own
} Panasonic drives so most people simply archive the images at our core and
} then move the ones they want by FTP as needed. I would like to switch to a
} more universal medium - namely CD ROM's. My understanding is that CD's can
} now be written to in multiple sessions so you don't need to fill an entire
} disk at once. Furthermore, it is my understanding that a disk of TIFF
} images should be readable by both IBM/WINTEL and Mac/PowerPC types
} computers. Is anybody actually doing this? Comments on how reliable are
} the recorders, which ones are best, pitfalls, etc would be appreciated.
} Before I get a dozen advocates of ZIP/Jazz drives, I don't want to go that
} route since that they are not as ubiquitous as CD drives. Thanks in
} advance.
}
}
} Thomas E. Phillips, Ph.D.
} Associate Professor of Biological Sciences
} Director, Molecular Cytology Core Facility
} 3 Tucker Hall
} University of Missouri
} Columbia, MO 65211
} (573)-882-4712 (voice)
} (573)-882-0123 (fax)


Dr. Lawrence F. Allard
Senior Research Staff Member
High Temperature Materials Laboratory
Oak Ridge National Laboratory
1 Bethel Valley Road
Bldg. 4515, MS 6064
PO Box 2008
Oak Ridge, TN 37831-6064

423-574-4981
423-574-4913 Fax
l2a-at-ornl.gov






From: Jinghua Tang :      jinghua-at-his.sb.fsu.edu
Date: Wed, 23 Jul 1997 22:56:39 -0400
Subject: help on CTF

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

I am a biophysics graduate student. One of my exam question
is how to correct CTF in image reconstruction. After finishing
the exam, I have more questions than answers!

What kinds of contrast exists in electron microscopic image
for weak-phase object? Is amplitude contrast the same as
aperture contrast?

What is the contribution of inelastic scattering on the image
contrast? How to account for it?

How does partial spatial coherence and temporal coherence
influence the image and electron diffraction?

Is it always sufficient to use just first order approximation
( the linear theory of phase and amplitude contrast image
formation ) for CTF consideration?

Is it true for low spatial frequencies ignoring the CTF was
better than compensating for phase contrast alone?

I guess compensation for the CTF was necessary and sufficient
to accurately reconstruct molecular densities. Could anyone tell
me the current ways for accurate determination of CTF?

By improving the different components in CTF, what is the optimal
contrast could be achieved in image?

Is it possible to put EM reconstruction on the same scale with
the structures derived from X-ray crystallography and NMR after
deliberate correction of CTF, solvent effects and differences between
atomic scattering factors for electron and X-ray?

To what extend we could use the insights obtained by building
models and comparison of the models with EM reconstruction?

Answers to any parts or aspects of these confusions I have will
be greatly appreciated!

Have a nice day!

Sincerely,

Jinghua Tang


--
{ {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} }

Mr. Jinghua Tang Phone: 850-644-4104(o)
Institute of Molecular Biophysics 850-574-9227(h)
Florida State University E-mail: jinghua-at-sb.fsu.edu
Tallahassee, FL 32306-4380 http://www.sb.fsu.edu/~jinghua
.....................................................................
The best way to have a good idea is to have lots of ideas.
-Linus Pauling
{ {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } {





From: jinghua-at-zen.sb.fsu.edu (Jinghua Tang)
Date: Wed, 23 Jul 1997 23:40:37 -0400
Subject: help on CTF

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


I am a biophysics graduate student. One of my exam question
is how to correct CTF in image reconstruction. After finishing
the exam, I have more questions than answers!

What kinds of contrast exists in electron microscopic image
for weak-phase object? Is amplitude contrast the same as
aperture contrast?

What is the contribution of inelastic scattering on the image
contrast? How to account for it?

How does partial spatial coherence and temporal coherence
influence the image and electron diffraction?

Is it always sufficient to use just first order approximation
( the linear theory of phase and amplitude contrast image
formation ) for CTF consideration?

Is it true for low spatial frequencies ignoring the CTF was
better than compensating for phase contrast alone?

I guess compensation for the CTF was necessary and sufficient
to accurately reconstruct molecular densities. Could anyone tell
me the current ways for accurate determination of CTF?

By improving the different components in CTF, what is the optimal
contrast could be achieved in image?

Is it possible to put EM reconstruction on the same scale with
the structures derived from X-ray crystallography and NMR after
deliberate correction of CTF, solvent effects and differences between
atomic scattering factors for electron and X-ray?

To what extend we could use the insights obtained by building
models and comparison of the models with EM reconstruction?

Answers to any parts or aspects of these confusions I have will
be greatly appreciated!

Have a nice day!

Sincerely,

Jinghua Tang

{ {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} }

Mr. Jinghua Tang Phone: 850-644-4104(o)
Institute of Molecular Biophysics 850-574-9227(h)
Florida State University E-mail: jinghua-at-sb.fsu.edu
Tallahassee, FL 32306-4380 http://www.sb.fsu.edu/~jinghua
.....................................................................
The best way to have a good idea is to have lots of ideas.
-Linus Pauling
{ {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } {




From: Stephen A. Shaffer :      sshaffer-at-microdataware.com
Date: Wed, 23 Jul 1997 23:26:21 -0700
Subject: Inter/Micro Day 3

Contents Retrieved from Microscopy Listserver Archives
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Arghhh!!! Sorry folks. Long day, banquet, yackin' with the gang. I'm
whupped. I'll have to include today with tomorrow's summary.

Steve Shaffer

p.s. Many have written to say thanks - You're welcome to all and thank
you back for the encouragement.





From: Benchaib Mehdi :      benchaib-at-rockefeller.univ-lyon1.fr
Date: Thu, 24 Jul 1997 13:08:10 +0200 (MET DST)
Subject: unsubscribe

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Dr Mehdi Benchaib
Laboratoire d' Histologie, Embryologie et Biologie de la Reproduction
8 avenue Rockefeller F-69373 Lyon CEDEX 08






From: John Minter :      minter-at-kodak.com
Date: Thu, 24 Jul 1997 07:35:06 -0400
Subject: Re: help on CTF

Contents Retrieved from Microscopy Listserver Archives
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Jinghua Tang wrote:

} I am a biophysics graduate student. One of my exam question
} is how to correct CTF in image reconstruction. After finishing
} the exam, I have more questions than answers!
} (snip - long list of questions deleted...)

You ask several good questions that space (and time) does not permit me to
answer in detail. May I suggest that you check out the Ph. D. Thesis of Xiadong
Zou (Electron Crystallography of Inorganic Structures - Theory and Practice.)
This was published in the Chemical Communications of Stockholm University (1995
No. 5.) You may be able to get a copy from your library. If not, write the
Department of Structural Chemistry, Arrhenius Laboratory, Stockholm University,
S-106 91 Stockholm, Sweden. Xiadong's thesis advisor was Professor Sven
Hovmoller.

In the first part of her thesis, Xiadong does a great job of summarizing the
theory of electron imaging and diffraction in a unified fashion. One of the
problems that we all face is that the theory was developed by several
communities and the terminology and notation is often conflicting and confusing.
For example, one needs to be careful to decide whether the author chooses
underfocus to be negative or positive. Xiadong shows several examples of the
effect of correction for CTF on image reconstruction.


--
Best Regards,
John Minter

Eastman Kodak Company Phone: (716) 722-3407
Analytical Technology Division FAX: (716) 477-3029
Room 2112 Bldg 49 Kodak Park Site email: minter-at-kodak.com
Rochester, NY 14562-3712 calendar: via PROFS






From: Woody.N.White-at-mcdermott.com
Date: Thu, 24 Jul 1997 8:26:00 -0500
Subject: Re: CD-ROM's for archiving-compatibility

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Haven't yet convinced the guys at work to spend the whopping {g}
$400-600 to implement a CD-R, but do have one on the home system
which works great for back-up/archiving. Is SCSI2 (aren't they
all?) running EzCD Pro software.

My HP 6020 does not support "packet mode" so it can't be treated
like a floppy, but does support "multi-session". It is not
necessary to write "disk at once". The caveat is that for each
writing "session" something like 13 to 20 Mb of CD space is
consumed as "overhead". ...But at $6 for 650Mb, do we care?

As to compatibility... some old CD players (under W3.1?) read the
first session and quit. When those systems were made, multisession
was not commonly available.... I haven't yet found a system old
enough that it would not read the CDs I burned.

Who really knows how long the CD-Rs will last (and will the
hardware be around to read them?). TDK rates their disks for 100
years....

For more CD-R info check:

http://www.cd-info.com/CDIC/Technology/CD-R/FAQ.html


Regards,
Woody White
Mcdermott Technology Inc.

Alt:
woody.white-at-worldnet.att.net
http://www.geocities.com/capecanaveral/3722
(Where I don't have enough space!)




From: :      kna101-at-utdallas.edu
Date: Thu, 24 Jul 1997 09:23:44 -0500 (CDT)
Subject: Re: GMA polymerization

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John,

If your only goal is to get thicker, larger sections with GMA, I
have a proceedure that doesn't require a special chamber or UV light. I
have used this procedure for 15 years with very good results. The blocks
stain well with toluidine blue, but little else, though. I use a modified
embedding procedure for JB-4, developed by Ann Klinker and Marge Hukee 15
years ago. It does require the blocks to be set without air, we use
parafilm sealed to the top of the mold. E-mail me if you are interested
in the details. kna101 utdallas.edu

Karen Pawlowski

On Wed, 23 Jul 1997, C. John Runions
wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Hi everyone, for many years I have been embedding in Spurr's resin but
} have recently decided to give glycol methacrylate a whirl for thicker LM
} sections of plant material. Many years have passed since my last work with
} GMA. I remember that we made a plexigalss container which we could
} evacuate the oxygen from by flushing with nitrogen and that polymerization
} was then carried out below a longwave UV light placed in the container.
} Does this sound correct? If so does anyone have any plans or tricks that
} might assist me in constructing such a device? Cheers, John
}
} =================
} C. John Runions, Ph.D
} Section of Ecology and Systematics
} Corson Hall
} Cornell University
} Ithaca, New York
} USA 14853
}
} email cjr14-at-cornell.edu [ie. cjr(fourteen)-at-...]
} phone (607) 254-4282
} Fax (607) 255-8088
}
}
}





From: Rick Cochran :      rick-at-msc.cornell.edu
Date: Thu, 24 Jul 1997 10:20:50 -0400 (EDT)
Subject: Re: CD-ROM's for archiving - any experience?

Contents Retrieved from Microscopy Listserver Archives
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John Hunt writes:
} From: John Hunt {hunt-at-msc.cornell.edu}
} Subject: Re: CD-ROM's for archiving - any experience? (fwd)
} To: rick-at-warf.msc.cornell.edu (Rick Cochran)
} Date: Thu, 24 Jul 1997 09:29:12 -0400 (EDT)
}
} I am forwarding this exchange from the microscopy listserver for
} your interest.

Which OS is best to run your CDR under depends on what environment your
date is being acquired in. The three environments which pop to mind are
Unix, Windows, and Mac. CDR solutions are available for all three
environments.

For our Unix environment, we have recently succeeded in setting up CDR
under Linux using freeware utilities. You need a SCSI system with a
suitably large disk, 'mkisofs' for mastering the CD images, and
'cdwrite' or 'cdrecord' for actually writing the CD. The only problem
we ran into was that most CDR drives require the 'SCSI disconnect'
feature which was disabled by default in the Linux SCSI driver for our
SCSI adapter.

mkisofs is available at tsx-11.mit.edu:/pub/linux/packages/mkisofs

cdwrite and cdrecord are available at sunsite.unc.edu:/pub/Linux/utils/disk-management

Since there is no industry standard for the SCSI commands for CDR
drives, support for each drive must be explicitly written into the
burning utility. You should determine which drive cdwrite and
cdrecord support before purchasing a drive.

Further information on CDR can be obtained from:

comp.publish.cdrom.hardware (most useful)
comp.publish.cdrom.software

http://www.cd-info.com/CDIC/Technology/CD-R/FAQ.html

Please don't ask me any questions.

-Rick

--
|Rick Cochran phone: 607-255-7223|
|Cornell Materials Science Center FAX: 607-255-3957|
|E20 Clark Hall, Ithaca, N.Y. 14853 email: rick-at-msc.cornell.edu|
| "The Founding Fathers did not establish the United States as a |
| democratic republic so that elected officials would decide trivia, |
| while all great questions would be decided by the judiciary." |
| Judge Andrew Kleinfeld |




From: Doug Cromey :      doug-cromey-at-ns.arizona.edu
Date: Thu, 24 Jul 1997 08:31:35 -0700
Subject: Re: normal versus digital darkroom

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Larry,

At 11:24 PM 7/23/97 -0400, you wrote:
} My question is: Has anyone *ever* gone back to access and use original
} image data that is 30 years old? I personally do not remember ever using
} original images that were even 5 years old....

Actually, I used to work in a Pathology diagnostic EM lab and we certainly
went back 20 years sometimes (rare diseases often require an entire career
to accumulate enough examples to publish about). I imaging the need for
archiving images is different for each field, but I still think its a bit
short sighted to not consider the future while living in the present.
Although this is off the science track, I wonder how our grandkids will
manage to view our digital snapshots when we're pretty sure that dye-sub
prints don't last that long and they need to find a viewer/reader for our
family album CD-ROM disk.

} Perhaps we should realistically not place too great an emphasis on the
} usefulness of saving all images for 30 years. CD-ROMs supposedly have that
} capability, but who believes that today's CDs will be readable by any
} technology available in 30 years? Anybody keep their old 8-track tape
} players in good service? I think most digital image formats will have to
} periodically be upgraded to the latest storage technologies, just as people
} take 16 mm home movies and convert them over to video tapes. Of course,
} this does not have to be done with film, and negatives probably will always
} be able to be converted easily to hard copy many years in the future....but
} I still never make images on film any more...

I may not have an 8-track (anymore), but how could we have the current crop
of re-releases of old musical works (originally recorded as analog) if the
music industry hadn't had a long lasting standard that they could still
work with today? Remember too that movie to video transfer results in a
lower resolution image (loss of information) stored on a tape format (I
hope you meant VHS, not the now obsolete BetaMax) that probably has a 5
year lifetime.

Actually, I'm of the mindset that of the mass storage technology currently
available today, CD-ROMs are the only ones likely to be readable in 30
years. That "Amazing Kreskin" like prediction (opinion) is based primarily
on the large market presence of CD readers (one industry analyst believes
that the installed base of CD devices will reach 150 million by the end of
1997, source: Advanced Imaging Magazine), compared with almost everything
else.

Don't get me wrong, I like digital imaging for most of the same reasons
everyone else likes it. I use it here at work and I try to teach students
and staff about it because I think its the way science is going. I'm just
concerned that we, as microscopists, are headed into this without realising
all the issues related to archiving of and longevity of images.

Yours,
Doug
.....................................................................
: Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy :
: Sr. Research Specialist 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




From: Stanley L. Flegler :      flegler-at-pilot.msu.edu
Date: Thu, 24 Jul 1997 12:22:21 -0400
Subject: RE: CD-ROM's for archiving/ZIP Drives

Contents Retrieved from Microscopy Listserver Archives
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Damian Neuberger mentioned that he had been told that ZIP drives were being
discontinued. I think his source of information must have been wrong.
Iomega recently introduced an internal SCSI version in addition to already
existing external parallel port and external SCSI. A number of computer
companies are now offering ZIP drives as standard equipment. We have used
ZIP drives for several years on this campus for storing images from TEMs,
SEMs, AFM/STMs, and Laser Confocal microscopes and we have been extremely
pleased with them. The disks are sold by our local Best Buy Store as well
as at most computer stores and Iomega has licensed other manufactures to
produce the disks. We also have the option of storing images on a
Panasonic Read/Write Magneto-Optical drive (1 GigaByte disks)on our SEM,
but most users prefer the ZIP. I for one don't like the idea of having a
thousand or so images on one disk. I much prefer to have several ZIP disks
with a hundred or so images on each.
Stanley L. Flegler, Assistant Director
Center for Electron Optics
Michigan State University





From: Jim Darley
Date: 24 July 1997 03:41
Subject: Re: Help requested for maladjusted glas

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Sharon

I can add little to what Jim Darley has said about setting up the knifemaker
but I was wondering whether you used to get good knives and now get bad or
have just started using the LKB.

Do you have the instruction charts for the knifemaker? Our LKB 7801A has a
glossy card 4 page manual and a condensed instruction laminated chart which
are very useful, when you understand them. They are normally hidden under
the machine on a little purpose built tray.

When you say that the reflection line is the wrong way do you mean that the
'meniscus' in the glass curves the wrong way? This might depend on which way
up you score your rhomboid/squares to make the triangles. We make 50 degree
knives from 100/80 degree rhomboids by first producing the rhomboids by
scoring on the roughened edge side of the glass strips. Then to bisect the
rhomboid we turn it over so all the rough edges are at the bottom. (I hope
you follow my meaning).

Finally what's your glass like - it hasn't been dropped or damaged. Is it
the right stuff for e.m. - we use 6mm thick glass from a reputable e.m.
supplier.

I hope this is of help but I'm sure that if you combine all of the replies
you will solve your problem.

Malcolm Haswell
e.m. unit
University of Sunderland
U.K.
----------

Sharon et.al.
That knife maker is an everlasting instrument but the adjustments must be
right, otherwise its a big waste of glass and time.
{SNIP}
----------------------------------------------------------------------------
-------------------------
} From: Sharon C. Thomas {sthomas-at-lanl.gov}
} To: Microscopy-at-sparc5.microscopy.com
} Subject: Help requested for maladjusted glass knifemaker
} Date: Wednesday, 23 July 1997 3:44
} The knifemaker available to me is a LKB Type 7801A from LKB Instruments
Inc.
} out of Rockville Maryland, but the company has since been taken over by
} Leica/Leo. The knifemaker has two wheel type gauges that adjust the
tension
} from opposing sides on the glass piece. I can make glass squares, but
when
} I try to cut the square into the two knives, I don't get the correct
shapes, even
} though I have tried systematically changing the settings on the two gauges
} many different ways. Typically, the "sharp" edge may be chipped, the
} reflection line in the glass is going in the opposite direction to what it
should
} and the opposite edge to the sharp edge may be either sharp as well or too
} thick a blunt edge. That's just one of the two knives; its' counterpart
often has
} two "sharp" edges. Can anyone with any direct experience with this model
of
} knifemaker provide me with any advice on how to adjust the settings to
produce
} two glass knives?
}
} Thanks in advance for any assistance that can be provided.





From: HILDEGARD CROWLEY :      hcrowley-at-du.edu
Date: Thu, 24 Jul 1997 12:04:06 -0600 (MDT)
Subject: G.Arentieri-no address-can't help

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I got a message from G. Arentieri asking for help. There was no return
address, so I cannot address him directly.
I can do a discussion of long term storage of tissue, but it is not of
general
interest to the group, so I would like to send it to G.Arentieri directly.
Greg, Could you try and contact me again? This time type your e-mail
address just in case of another snafu.
Bye,
Hildy




From: kjd136-at-email.psu.edu (kelly dowhower)
Date: Thu, 24 Jul 1997 14:38:26 -0400
Subject: Difficulty detecting antibody staining in transfected cells

Contents Retrieved from Microscopy Listserver Archives
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I have been transfecting mammalian cells with a dopamine receptor and
trying to detect, via fluorecence microscopy, its expression with a
polycolonal antibody that I made to one of this dopamine receptor's
intracellular loops. (The antibody detects the dopamine receptor epitope,
to which the antibody was made, on western blots, when expressed by E. coli
in the context of a fusion protein, so I know the antibody does recognize
the epitope against which it was made.)

When I express the receptor in transfected HEK293 cells, fix the cells
by methanol:acetone, and do fluorescence microscopy, I get no detection of
receptor expression. I engineered a " FLAG tag" into the receptor and
stained for FLAG labeling;the FLAG epiope gave intense staining, so I know
the dopamine receptor is being expressed.

Does anyone have any thoughts on what factors I might to alter in my
immunofluorescence assay in order to make the dopamine receptor antibody
recognize the dopamine receptor in the transfected cells? It seems as if
the receptor is folded in a conformation such that the epitope that the
antibody recognizes is buried / not exposed. Are there better/more
appropriate fixation methods? Does anyone have information on live
staining or microwave heating/steaming of the cells?

Thanks for your input.

Kelly Karpa
kjd136-at-psu.edu






From: John F. Mansfield :      jfmjfm-at-umich.edu
Date: Thu, 24 Jul 1997 15:53:01 -0400
Subject: Summer Issue of MicroNews now on Web

Contents Retrieved from Microscopy Listserver Archives
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Hello there,
MAS Members and other interested parties,
The latest issue of The Microbeam Analysis Society Newsletter, MicroNews
(Summer 97), is now available on the Web at:

http://www.microanalysis.org/mas/masmn/micronews97/mnsummer97.html

Take a look.

Regards,

John Mansfield





John Mansfield
North Campus Electron Microbeam Analysis Laboratory
417 SRB, University of Michigan
2455 Hayward, Ann Arbor MI 48109-2143
Phone: (313) 936-3352 FAX (313) 936-3352
Cellular Phone: (313) 715-2510
(Leaving a phone message at 936-3352 is preferable to 715-2510)
Email: jfmjfm-at-engin.umich.edu
URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html






From: Greg :      greg-at-umic.sunysb.edu
Date: Thu, 24 Jul 1997 16:04:34 -0400
Subject: Hummer VI-a sputter coater

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Dear all,
I need some information. I have a Hummer VI-a sputter coater and I am
having trouble finding where to get targets. EMCORP which sold the
instrument seems to be out of business. Their phone numbers do not work.
Any help will be appreciated.

TIA
Greg Rudomen
University microscopy Imaging Center
S.U.N.Y. -at- Stony Brook




From: Peter Michiels :      pmi-at-mail2.tornado.be
Date: Thu, 24 Jul 1997 23:02:57 +0100
Subject: unsubscribe

Contents Retrieved from Microscopy Listserver Archives
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Please unsubscribe me.
___________________________________

Ing. Peter Michiels

Philips Analytical - Electron Optics
Tel.: +32-2-5256249
Fax: +32-2-5256483
e-mail: pmi-at-tornado.be
___________________________________





From: HILDEGARD CROWLEY[SMTP:hcrowley-at-du.edu]
Date: Thu, 24 Jul 1997 17:16:08 -0400
Subject: G.Arentieri-no address-can't help

Contents Retrieved from Microscopy Listserver Archives
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Gregory.Argentieri-at-sandoz.com

This is the address i have on file for Greg


----------


I got a message from G. Arentieri asking for help. There was no return
address, so I cannot address him directly.
I can do a discussion of long term storage of tissue, but it is not of
general
interest to the group, so I would like to send it to G.Arentieri directly.
Greg, Could you try and contact me again? This time type your e-mail
address just in case of another snafu.
Bye,
Hildy







From: Geoff McAuliffe :      mcauliff-at-UMDNJ.EDU
Date: Thu, 24 Jul 1997 16:21:06 -0700
Subject: Tissue storage by Hildy

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Dear List:

I for one would be very interested in Hildy Crowley's opinions on long
term tissue storage. Others ...?

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 e-mail: mcauliff-at-umdnj.edu
***************************************************************




From: Robert Underwood :      underwoo-at-u.washington.edu
Date: Thu, 24 Jul 1997 15:29:41 -0700 (PDT)
Subject: Re: Difficulty detecting antibody staining in transfected cells

Contents Retrieved from Microscopy Listserver Archives
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Howdy,

The first thing I was wondering: Is the epitope extra or intra cellular?
I guess the most success I've had is by manipulating the fixation. Trying
the solvents vs. 2-4% paraformaldehyde or something like Zambonies or
Bouins so the picric acid can go in fast and stablize the protien then the
paraformaldehyde lightly crosslinks. Then if you need to open up the cell
I use .1% Tween or to open epitopes a very short 1- 3min .01% Trypsin.

Bob
Morphology Core


On Thu, 24 Jul 1997, kelly dowhower wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} I have been transfecting mammalian cells with a dopamine receptor and
} trying to detect, via fluorecence microscopy, its expression with a
} polycolonal antibody that I made to one of this dopamine receptor's
} intracellular loops. (The antibody detects the dopamine receptor epitope,
} to which the antibody was made, on western blots, when expressed by E. coli
} in the context of a fusion protein, so I know the antibody does recognize
} the epitope against which it was made.)
}
} When I express the receptor in transfected HEK293 cells, fix the cells
} by methanol:acetone, and do fluorescence microscopy, I get no detection of
} receptor expression. I engineered a " FLAG tag" into the receptor and
} stained for FLAG labeling;the FLAG epiope gave intense staining, so I know
} the dopamine receptor is being expressed.
}
} Does anyone have any thoughts on what factors I might to alter in my
} immunofluorescence assay in order to make the dopamine receptor antibody
} recognize the dopamine receptor in the transfected cells? It seems as if
} the receptor is folded in a conformation such that the epitope that the
} antibody recognizes is buried / not exposed. Are there better/more
} appropriate fixation methods? Does anyone have information on live
} staining or microwave heating/steaming of the cells?
}
} Thanks for your input.
}
} Kelly Karpa
} kjd136-at-psu.edu
}
}
}





From: Annette Bakker Inserm U153 :      abakker-at-myologie.infobiogen.fr
Date: Thu, 24 Jul 1997 18:12:17 -0500
Subject: which inverted microscope?

Contents Retrieved from Microscopy Listserver Archives
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Hi,
we would like to buy an inverted and fluorescent microscope in our
institute in Paris






From: Lucille A. Giannuzzi :      lag-at-pegasus.cc.ucf.edu
Date: Thu, 24 Jul 1997 19:17:02 -0400
Subject: SEM backscattered kikuchi patterns

Contents Retrieved from Microscopy Listserver Archives
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Hello all:

I have an undergraduate student working on the design of a homemade
backscatter diffraction pattern for the SEM. We are trying to get patterns
using a JEOL T300. We tried using an accelerating voltage of up to 30keV,
turned up the spot size, turned down the gun bias, and we get a "bright"
screen but no kikuchi lines. We tried different specimen tilts, specimen
working distances, and screen to specimen distances, and the specimen is a
good quality Si wafer, but nothing seems to produce a bright enough signal
to generate the Kikuchi lines. Are we limited by the SEM we are using, or
are we just missing something?

Thanks.
Lucille Giannuzzi


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

Dept. of Mechanical, Materials, and Aerospace Eng.

University of Central Florida phone (407) 823-5770
PO Box 162450 fax (407) 823-0208
4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu
Orlando, FL 32816-2450 USA
*************************************************************************







From: :      yoyodine-at-UNM.EDU
Date: Thu, 24 Jul 1997 18:01:53 -0600 (MDT)
Subject: Re: CD-ROM's for archiving-compatibility

Contents Retrieved from Microscopy Listserver Archives
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On the subject of archiving:

Zip drives are not yet discontinued...but they probably have a shorter
market life-span than CD-R's (hard to predict the future though).

Optical drives are neat for archiving...but who has an optical drive on
thier home/office system?? They are rather pricy.

CD-R's are the way to go I believe. Almost all computers these days have
CD devices. The Disk and the drives are rather inexpensive. The storage
life of a CD (not in vacuum) is over 60 years (and far longer in Vacuum
storage). We store Tiff files and use HTML frontends so that almost
ANYONE with ANY SYSTEM with ANY SOFTWARE can use them (OK...maybe
not...but I have heard of no problems so far). They just make sence. We
do not use a dedicated machine...But I would suggest to anyone that they
at least use a dedicated h-drive for writing to (helps alot). I guess if
a lab does not allow alot of access to archives then maybe an Optical
drive would be better (less need for compatability).

On the subject of archiving medium going out of date:

At first thought one might think this is a horrible problem. We have
about 200 old 8 inch floppies and no drive to read them. Further...I am
not so sure how many of them are any good. They contain alot of years of
data.

Yet, and I know this may not apply to everyone, what is the data worth??
I have found that the archiving Medium and devices seem to out last the
Data. Any Data around here that are more than about 10 years old are
almost worth-less. The machines used to collect them are long since
replaced with better machines...we would have to recollect any archived
data that old. Further, almost all older data worth note has been
published and exists in hard copy some place.

Its neat to think that 100 years from now, someone some place will want to
see some of my data, or a perfect BSE image I took. When that thought
comes to mind I get a bit excited...then suddenly a new thought appears,
"Yeah Right Christopher!! Keep Dreaming!". Think about the machines that
will be in use 100 years from now.

Christopher






From: Stephen A. Shaffer :      sshaffer-at-microdataware.com
Date: Thu, 24 Jul 1997 20:22:43 -0700
Subject: Inter/Micro Day 4

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Inter/Micro 97 Day 4

I'll start with some notes on Wednesday's sessions since I neglected to
summarize them last night. I was, er, a bit slow by the time I got
back. :-P The session focus on Wednesday was "History and Art,"
although that was only loosely held to. Several of the papers dealt
with artistic subjects but not art conservation per se.

Gary Laughlin spoke about "A Unique Metallurgical Process From the Early
Bronze Age" in which he described the findings at, and significance of,
a site excavated at the Kestel Mine in the Taurus Mountains of Turkey.
The site dates from the third millenium BC. The examinations indicate
that cassiterite ore was mined and refined at the site to yield a black
magnetic oxide. This would have been more readily separated, due to its
magnetism, than could be accomplished by other means.

Dr. McCrone gave an "Update on the Turin Shroud" in which he reviewed
several letters he received from Father Rinaldi prior to the Father's
death in 1993. In the letters, Father Rinaldi effectively acknowledged
the proof that the Shroud was painted and actually dated from much later
than the time of Christ's crucifiction, thus was not the true article.

(For those of you who may not know, Dr. McCrone concluded early on in
the Shroud investigations, and from microscopic observations alone, that
the shroud was a painting. He stood nearly alone in this view and was
vilified for nearly two decades before Carbon dating ultimately
confirmed his conclusions.)

John Delly gave one of his typical, amazing presentations, this one on
"Hand-colored Microscopical Illustrations." In it John took his
audience on a delightful stroll through 19th century microscopy
publications illuminated with hand-colored illustrations. He
demonstrated the evolution and later de-evolution of the quality of such
illustrations, the variation that one can see from different
illustrators of the same work, and the differences that are seen
edition-to-edition of the same work. Of course, when he became
interested in the subject, John felt compelled to master the art
himself. Through his own study and practice he gained the insignt
necessary to understand and explain the techniques and variations seen
in these historical works. The illustrations are, indeed, beautiful and
many of us are fortunate to own examples of these illustrations in early
works on microscopy. Because of the vast number of color illustrations
necessary to address his subject, it is unlikely that this presentation
will ever be recorded fully in print. (How about a book, John?) Those
of us fortunate enough to be in the room may be the only ones ever to
enjoy this particular product of John's efforts. Thank you, John, once
again.

Have you ever stood befor an audience wondering why on earth you found
yourself presenting in the particular session where you were? Wayne
Moorehead must have when he spoke on "A Tale of Two Danas; Influences in
Mineralogy" but he soldiered on and did a fine job chronicaling the
lives of two remarkable men. The mineralogists in the audience will
need no introduction to the Danas but I'll just mention for the others
that the elder Dana published his first edition of the System of
Mineralogy in 1835 at the tender age of 24. It was the first such major
scientific work of classification written in English and he and his son
went on to publish or edit a vast array of classic works in Mineralogy.
Together or individually, they edited the prestigious American Journal
of Science continuously for an astounding 95 years, from 1840 to 1935.
One of the most noteable achievements that Wayne mentioned, in my mind
anyway, was when James Dana, in the introduction to a revised edition of
his classic System, abruptly abandoned his entire earlier classification
system as outdated!. Believing that system no longer consistant with
emerging thought, he just as abruptly adopted and described a newer
system which largely stays with us today. I find such honest
self-appraisal and the ability to continue to move forward without
missing stride quite refreshing.

Wednesday afternoon was occupied by two sessions which would be unusual
at other meetings. Using video microscopy, Anna Teetsov of McCrone
Associates demonstrated some micromanipulation techniques within the
context of creating artistic works on microscope slides by arranging
butterfly scales of various colors into micro-images. Anna and a few
others continue to develop this art form which is particulary unique to
the community of microscopists. One has to have a microscope and
micro-related knowledge and skills in order to produce these beautiful
little creations, then one has to have a microscope to view them as
well. Kind of nice, don't you think? Something we can hold purely for
the aesthetic pleasure and uniquely our own.

The afternoon was closed with a demonstration by Alan Shin on how one
can construct a working replica of Leeuwenhoek's single lens
microscopes. I did not attend this demonstration as I have on a
previous occasion taken a longer version from Alan in which we got to
actually construct our own microscopes. Comments from those who did
attend and look through the instrument Alan made reflected surprise at
how much one could see and pleasure at the experience of seeing an
insturment of such historical significance actually fabricated.

Today's sessions were on Forensic Microscopy. Jose Almirall told us
about "Developments in Glass Examination: Automated Microscopy
Techniques and Composition Analysis." Jose's talk was very interesting
and perhaps somewhat troubling to practicing forensic scientists as he
told us of (among other things) a remarkable consistancy in the optical
properties of some glasses, especially window glass manufactured by the
"float" process. The new information for me was the time over which the
products of these plants will remain indistinguishable under
conventional forensic examination techniques. I am not aware of other
time-dependant studies of glass properties but Jose showed data
collected over at least 18 months, during which the product of a float
glass plant was entirely uniform in refractive index. He did however,
offer a remedy for this disturbing finding. He showed that glass
samples which are indistinguishable by refractive index can often be
distinguished by elemental analysis of Fe, Mg, Al, and Zr using
ICP/AES. Now all the forensic people have to do is get themselves one
of these and... ;-)

Wayne Moorehead gave another excellent paper on Thursday, this one on
"An Introduction to Microscopical Feather Identification." Wayne told
us that the flight and tail feathers of birds are not always useful for
identification but that the down or contour (breast) feathers can be
distinctive, at least down to the order of birds, occasionally to the
family, but virtually never to the genus or species. Still,
identification at this level may prove very useful in a forensic case.
Wayne illustrated how appropriate preparations can be made, what
features of the feather barbule to examine and how they can vary. He
also showed and described the identifying characteristic of numerous
feather types.

Thom Hopen gave an interesting talk on "Teaching Forensic Microscopy in
Countries Formerly Known as the Soviet Union." Thom responded to a
State Department request that he make numerous trips to various
countries of the former Soviet Union. He has taught courses of fiber
and paint comparison, explosives residue analysis, and basic
microscopy. He found his students to be highly motivated and dedicated
people, anxious for quality instruction in basic forensic microscopy
techniques. Often they are at least adequately equiped though sometimes
have little or no idea how to fully exploit the equipment they have.
(Unfortunately, when it comes to microscopy, this is too often true here
also! My comment, not Thom's.) One can only immagine the difficulty of
teaching in a completely and literally foreign environment, working
through a translator, and using instrumentation previously never seen.
Often, Thom had to set the equipment in proper working order prior to
beginning instruction. But apparently all has worked out for him and
his students and several more trips are planned to continue the
education.

Well folks, I think I'll stop there. Of course, there were many more
fine presentations and, once again, I'll mention that my choice of
topics covered here in no way reflects badly on the other papers. All
of the presentations were excellent.

Tomorrow is given over to a tutorial workshop on the Dispersion Staining
technique. It will be given at McCrone Research Institute by Dr.
McCrone and will be attended by twenty-odd students, all that can
reasonably be accomodated in such a hands-on session. For the rest of
us, this afternoon marked the end of another educational, enlightening,
and just plain fun Inter/Micro.

Special thanks, as usual, to Nancy Daerr who coordinates all
arrangements for these meetings and who, as usual, did an exceptional
job of taking care of us and making our stay wholly enjoyable.

To all of those interested in these meetings, please note: Next year
marks the Golden Anniversary of Inter/Micro, the fiftieth anniversary.
(Wow!) Plan on attending what promises to be an excellent meeting.
Contact Nancy Daerr for further information, to be put on a mailing
list, etc. She can be reached at McCrone Research Institute, 2820 S.
Michigan Avenue, Chicago, IL, 60616 or simply as ndaerr-at-mcri.org. The
phone numbers at McRI are 312-842-7100 (voice) and 312-842-1078 (fax).

It's been a pleasure being your ears at Inter/Micro 97. But
tomorrow... Ahhh, Chicago! The architecture, the museums, the Art
Institute! I feel like a nice walk! Happy trails to all, and to all,
Good Night.

Steve Shaffer
MicroDataware
sshaffer-at-microdataware.com





From: js_vetrano-at-ccmail.pnl.gov (John S Vetrano)
Date: Thu, 24 Jul 1997 15:16:42 -0700
Subject: TEM Position Announcement

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Greetings all.

Please post or pass this on as appropriate. Thank you.

Regards,

John Vetrano

----------

Electron Microscopy Positions
in Materials Characterization

Pacific Northwest National Laboratory

The Structural Materials Research Group at Pacific Northwest National Laboratory
is seeking applicants for a staff position and a post-doctoral appointment, both
specializing in the characterization of materials using transmission electron
microscopy (TEM). The Electron Microscopist staff position requires a minimum
of a 2-year specialized degree with expertise and training in the operation of
analytical TEM instruments as well as in the preparation of electron transparent
materials for examination. This individual will conduct research on a wide
variety of metallic, ceramic and composite materials in support of scientists
and engineers in the Structural Materials Research Group. Current activities
include both basic and applied research in the areas of deformation mechanisms
in metals, interfacial segregation and precipitation, radiation effects in
metals, ceramics and composites, and enviromental degradation. In addition, the
position will help oversee the maintainance of the Group microscopy facilities
which include three conventional TEMs, a field-emission-gun (FEG) TEM and a
scanning TEM, SEMs and a scanning Auger microprobe.

A post-doctoral position is also available focussed on the high-resolution
characterization of grain boundaries in metallic alloys by analytical TEM.
Energy dispersive x-ray and electron energy loss spectroscopies will be used
with the FEG-TEM to elucidate segregation and precipitation mechanisms in
aluminum and stainless steel alloys. The initial post-doctoral position would
be for one-year and a second-year renewal is possible. The position requires a
PhD degree in material science, physics or a related discipline and experience
in analytical TEM for quantitative compositional analysis.

Pacific Northwest National Laboratory (PNNL) is operated by Battelle Memorial
Institute for the U.S. Department of Energy. PNNL is a multiprogram laboratory
located at the Hanford site near the junction of the Columbia, Snake and Yakima
rivers in SE Washington state.

For consideration, please submit a resume (including references) and publication
list by Oct. 1, 1997 to: Dr. Stephen M. Bruemmer, Pacific Northwest National
Laboratory, Battelle Boulevard, P.O. Box 999, Richland, WA 99352. Phone:
(509) 376-0636; Fax: (509) 376-6308; e-mail: sm_bruemmer-at-pnl.gov.




From: Wolf in Melbourne :      wschweitzer-at-access.ch
Date: Fri, 25 Jul 1997 15:14:58 +1000
Subject: CD-ROM's for archiving - any experience?

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Using different Apple Macintosh Workstations and one mobile Apple
Powerbook 1400cs (which has an 8xCD-ROM drive), the CD-R Backup solution
proved perfect for Software and Data Backup, and taking complete backup
with the Powerbook still leaves me very mobile.

I use the Philips 2660 CD-Burner with Astarte Toast CD Software. This
not only allows for different formats, but also for creation of CD-ROM
image files on harddisk for easy preparation of a 650M partition, or
smaller. If the Macintosh is setup well, and the driver software is
properly installed (I use the FWB-CD-driver although not recommended by
Astarte), no problems are expected to occur.

To deal with the fact, that some older machines do not read multi
session CD-ROMs, I installed a 1G-Harddisk to collect the data to be
burned on the CD on a 650M image file. As soon as it is full, I burn 2
CDs (one for regular use, one for backup), and throw the image file
away. This proved itself useful, because at the time I burn the CD, some
files are no longer needed and can be thrown away before burning.

A known fact with Floppy Disks is the time they keep the data until they
begin to loose it (1-2 years? depending on brand?). This issue might be
important and also brand-dependent in CDs as well, but the extent, the
expected timerange and hence significane is unknown to me.

Wolf Schweitzer, MD
Research Fellow, VIFM, Melbourne, Australia
wschweitzer-at-access.ch




From: roadwalk-at-sprynet.com
Date: Fri, 25 Jul 1997 04:44:12 -0700
Subject: Re: CD-ROM's for archiving - any experience?

Contents Retrieved from Microscopy Listserver Archives
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This listserver is a great help to me as an amature microsopist. I enjoy the
information obtained by reading the daily "chat". I work 8 hours a day in the
computer business. My suggestion is never trust any technology of today to last
very long. The abiliity to store files for a long time is worthless if the
technology on which it is stored is antiquated by as much as 3 - 5 years.

Zip drives are hot technology right now, but becauase of their limitations of
size, at this point, they are being passed by with upcoming technology. File
storage is good for the time being, but in less than a year look for the most
modern and up to date file storage technology to change. Computers and their
technology are always changing and becoming less expensive as the R&D is paid
off and manufacture of newer ideas comes into being.

Do not hitch your wagon to anything as a forever thing. If silver grain
photographs are s "thing of the past", whatever we use today will be also, very
soon. CDRom drives and platters are fantastic, but technology to handle 1000
times the information storage in a smaller package via opti-digital technology
is somewhere close around the corner.

I am not a proponent of either silver grain or digital file storage. Know that
your will have to provide the latest thing for your business client there is and
that will always change. Two years ago 4X CDRom drives were all the rage.
Today you have to invest in a 16X or 20X drive that is less expensive than the
4X was. 4 gigbit hard drives retail for less than half the cost of a 540
megabit drive 5 years ago.

TAKE A LESSON!!!!!

TIME MARCHES ON [SO DOES TECHNOLOGY]




From: Woody.N.White-at-mcdermott.com
Date: Fri, 25 Jul 1997 8:11:00 -0500
Subject: CD-R archiving time...

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FWIW...

Sometimes it dosen't matter if the data will actually ever be
accessed. It must just be available. Certain regulations (QA type
stuff) can require data archiving for 30 years or more.

Woody White
Mcdermott Technology Inc.




From: Woody.N.White-at-mcdermott.com
Date: 7/24/97 2:59 PM
Subject: Hummer VI-a sputter coater

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Any target the correct diameter will work. What I have done is clean-up
the
spent target assembly, then silver-epoxy a new (generic) disk to the target
assembly face. Most EM suppliers sell disks. Material can also be
purchased
from suppliers like Alfa Aesar (Johnson Matthey) or Goodfellow. There is a
supplier in Nevada (I think) who is reputed to be less expensive, but I
don't
have any more info available...Maybe someone else knows that name/address.

Woody White
Mcdermott Technology Inc.

Alt:
woody.white-at-worldnet.att.net
http://www.geocities.com/capecanaveral/3722
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Dear all,
I need some information. I have a Hummer VI-a sputter coater and I am
having trouble finding where to get targets. EMCORP which sold the
instrument seems to be out of business. Their phone numbers do not work.
Any help will be appreciated.

TIA
Greg Rudomen
University microscopy Imaging Center
S.U.N.Y. -at- Stony Brook




From: leibest-at-acpub.duke.edu (Leslie Eibest)
Date: Fri, 25 Jul 1997 08:17:15 -0500
Subject: Re: Hummer VI

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X-Sender: leibest-at-mail-le.acpub.duke.edu
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Greg;
I get targets for my Hummer V sputter coater from:
Anatech, Ltd.
5510 Vine St.
Alexandria, VA 22310
(703) 971-9200

Apparently, this company is owned by former Hummer management and
employees, so they can probably help you.

Leslie Eibest
Zoology Dept., Box 90325
Duke University
Durham, NC 27708 USA
(919) 684-2547
leibest-at-duke.edu






From: John Arnott :      ladres-at-worldnet.att.net
Date: Fri, 25 Jul 1997 08:25:02 -0400
Subject: Re: Hummer VI-a sputter coater

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Greg wrote:
}
} ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.}
} Dear all,
} I need some information. I have a Hummer VI-a sputter coater and I am
} having trouble finding where to get targets. EMCORP which sold the
} instrument seems to be out of business. Their phone numbers do not work.
} Any help will be appreciated.
}
} TIA
} Greg Rudomen
} University microscopy Imaging Center
} S.U.N.Y. -at- Stony Brook

Greg,
We supply targets for all models of Hummer sputter coaters and almost
any other type you might need. Please let me know what material and I
will send you a quote.
Thanks,
JD Arnott
Ladd Research




From: John Arnott :      ladres-at-worldnet.att.net
Date: Fri, 25 Jul 1997 08:24:02 -0400
Subject: Re: Hummer VI-a sputter coater

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Greg wrote:
}
} ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.}
} Dear all,
} I need some information. I have a Hummer VI-a sputter coater and I am
} having trouble finding where to get targets. EMCORP which sold the
} instrument seems to be out of business. Their phone numbers do not work.
} Any help will be appreciated.
}
} TIA
} Greg Rudomen
} University microscopy Imaging Center
} S.U.N.Y. -at- Stony Brook

Greg,
We supply targets for all models of Hummer sputter coaters and almost
any other type you might need. Please let me know what material and I
will send you a quote.
Thanks,
JD Arnott




From: Bob_Citron-at-cc.chiron.com (Bob Citron)
Date: 7/24/97 4:04 PM
Subject: Hummer VI-a sputter coater

Contents Retrieved from Microscopy Listserver Archives
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Greg:

Have you already tried Anatech?

Anatech Ltd.
6621-F Electronic Dr.
Springfield, VA 22151
Ph: 800-Plasma-9

Regards,

Bob Citron
Chiron Vision
Claremont, CA
(909) 399-1311
Bob_Citron-at-cc.chiron.com


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Dear all,
I need some information. I have a Hummer VI-a sputter coater and I am
having trouble finding where to get targets. EMCORP which sold the
instrument seems to be out of business. Their phone numbers do not work.
Any help will be appreciated.

TIA
Greg Rudomen
University microscopy Imaging Center
S.U.N.Y. -at- Stony Brook




From: Owen P. Mills :      opmills-at-mtu.edu
Date: Fri, 25 Jul 1997 10:45:40 -0400
Subject: Contrast in silicon dendrites

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Message-Id: {3.0.1.32.19970725104540.007717d4-at-mmserver.mm}
X-Sender: opmills-at-mmserver.mm
X-Mailer: Windows Eudora Pro Version 3.0.1 (32)

Hello,

I'm posting this query for a colleague here at MTU. Please contact him
directly at qchorn-at-mtu.edu. Thanks.

Owen

++++++++++++++++++++++++

While characterizing the microstructure of a commercial ferrosilicon alloy,
an interesting contrast event was observed within the silicon phase. The
center of the silicon dendrites appears brighter than the edges when
secondary electron imaging is used. This contrast is not observed using
backscattered electron imaging. The following web site has images showing
this contrast as well as other information about the alloy being examined:

http://www.mm.mtu.edu/~qchorn/siliconquestion.htm

Any insight as to what could be causing this contrast would be greatly
appreciated.

Thanks,

Quinn C. Horn
++++++++++++++++++++++++++
Owen P. Mills
Michigan Technological University
Metallurgical & Materials Engineering
Rm 512 MME Building
Houghton, MI 49931
906-487-2002
906-487-2934 FAX
opmills-at-mtu.edu




From: Bob_Citron-at-cc.chiron.com (Bob Citron)
Date: 7/24/97 6:01 PM
Subject: Re: CD-ROM's for archiving-compatibility

Contents Retrieved from Microscopy Listserver Archives
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Hi Christopher;

I agree with your comments that we sometimes try to save "all" of the data
we generate, and much of it is of little future value. But having worked
in the medical device industry for 20 years now, I would like to inject my
two cents, just to provide another viewpoint.

In industry, some data (regardless of its age) is absolutely invaluable.
An example would be data that is generated to support studies of the
safety/efficacy of an implantable device. (Silicone breast implants are a
good example). If 20 years from now, litigation arises with regard to one
of our products, raw data may be required to lend credence to our studies.
In fact, the FDA requires that we maintain certain data "forever", and some
of this data is electronic. With this in mind, I have wrestled with this
topic of data archiving and integrity for several years now. I try to
ensure future compatibility of whatever software and hardware upgrades I
make in our data collection equipment, but it's not always possible.
Consequently, I have made it a point to also archive some of the older
hardware and software, just in case.

Best Regards,

Bob
**********************************
Bob Citron
Chiron Vision
Claremont, CA
USA
(909)399-1311
Bob_Citron-at-cc.chiron.com
**********************************


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On the subject of archiving:

Zip drives are not yet discontinued...but they probably have a shorter
market life-span than CD-R's (hard to predict the future though).

Optical drives are neat for archiving...but who has an optical drive on
thier home/office system?? They are rather pricy.

CD-R's are the way to go I believe. Almost all computers these days have
CD devices. The Disk and the drives are rather inexpensive. The storage
life of a CD (not in vacuum) is over 60 years (and far longer in Vacuum
storage). We store Tiff files and use HTML frontends so that almost
ANYONE with ANY SYSTEM with ANY SOFTWARE can use them (OK...maybe
not...but I have heard of no problems so far). They just make sence. We
do not use a dedicated machine...But I would suggest to anyone that they
at least use a dedicated h-drive for writing to (helps alot). I guess if
a lab does not allow alot of access to archives then maybe an Optical
drive would be better (less need for compatability).

On the subject of archiving medium going out of date:

At first thought one might think this is a horrible problem. We have
about 200 old 8 inch floppies and no drive to read them. Further...I am
not so sure how many of them are any good. They contain alot of years of
data.

Yet, and I know this may not apply to everyone, what is the data worth??
I have found that the archiving Medium and devices seem to out last the
Data. Any Data around here that are more than about 10 years old are
almost worth-less. The machines used to collect them are long since
replaced with better machines...we would have to recollect any archived
data that old. Further, almost all older data worth note has been
published and exists in hard copy some place.

Its neat to think that 100 years from now, someone some place will want to
see some of my data, or a perfect BSE image I took. When that thought
comes to mind I get a bit excited...then suddenly a new thought appears,
"Yeah Right Christopher!! Keep Dreaming!". Think about the machines that
will be in use 100 years from now.

Christopher






From: Bob_Citron-at-cc.chiron.com (Bob Citron)
Date: Fri, 25 Jul 1997 09:04:10 -0700
Subject: Inter/Micro Summaries

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Hi All;

I just want to publicly thank Steve Shaffer for the interesting summaries
of the talks at Inter/Micro. I hope I speak for all when I say that those
of us who (for whatever reason, be it time, money, other committments)
could not attend, really appreciated it. Hope you enjoyed the "off" time
as well!

Best Regards,

Bob
**************************
Bob Citron
Chiron Vision
Claremont, CA
USA
Bob_Citron-at-cc.chiron.com
**************************




From: jmkrupp-at-cats.ucsc.edu (Jon Krupp)
Date: Fri, 25 Jul 1997 09:53:12 -0700
Subject: Dye sub printer leftovers

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Hi:

Has anyone thought of something useful to do with all the leftovers from
dye sub printers? I have boxes of used ribbons and other things that seem
too good just to toss out. Could they be used for something, or maybe
recycled somehow?

Jonathan Krupp
Microscopy and Imaging Lab
University of California
Santa Cruz, CA 95064
(408) 459-2477
FAX (408) 429-0146
jmkrupp-at-cats.ucsc.edu






From: bauer%wp94.ferro%ferro1ge#-at-ferro.geis.com
Date: Fri, 25 Jul 97 19:01:00 GMT
Subject: M&M '97 - Indians tickets

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Hi everyone,
I have a couple of announcements about tickets to the Cleveland Indians
game on Tues evening.
One - If you have reseved tickets, I need to see the money!!
Please send a check made out to 'MSA' for $20/ticket (cheap seats, but
includes a picnic) to me here at:

Ferro Corp.
7500 E. Pleasant Valley Rd.
Independence, OH 44131

Two - If you plan on canceling your reservations, I've had a number of
requests for more tickets...So - I'm starting a waiting list. First come, first
on the list. I have had a few cancelations, so far - so please give me a
call at (216)641-8585 x6613. E-mail address is: vbauer-at-ferro.geis.com.

I know this isn't "Microscopy" but I don't have current e-mail addresses
for everyone, so please bear with me.

Thanks a lot!!
Vicky (now Bryg)




From: tom moninger :      moninger-at-emiris.iaf.uiowa.edu
Date: Fri, 25 Jul 1997 16:32:41 -0500 (CDT)
Subject: Re: Dye sub printer leftovers

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One woman that used to work in our lab thought the ribbons would be good
for Halloween--go to a party as the "Primary Colors"

Thomas Moninger moninger-at-emiris.iaf.uiowa.edu
University of Iowa Central Microscopy Research Facility
http://www.uiowa.edu/~cemrf
Views expressed are mine.
On Fri, 25 Jul 1997, Jon Krupp wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Hi:
}
} Has anyone thought of something useful to do with all the leftovers from
} dye sub printers? I have boxes of used ribbons and other things that seem
} too good just to toss out. Could they be used for something, or maybe
} recycled somehow?
}
} Jonathan Krupp
} Microscopy and Imaging Lab
} University of California
} Santa Cruz, CA 95064
} (408) 459-2477
} FAX (408) 429-0146
} jmkrupp-at-cats.ucsc.edu
}
}





From: Ann Rushing :      Ann_Rushing-at-BAYLOR.EDU
Date: Fri, 25 Jul 1997 12:47:16 -0500
Subject: Philips 201 TEM

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Subject: Time:12:45 PM
OFFICE MEMO Philips 201 TEM Date:7/25/97

If anyone is interested in obtaining a Philips 201 (24 years old, on service
contact continually) for a nominal cost, please contact me immediately.
Ann E. Rushing
Department of Biology
Baylor University
Waco, TX 76798
Ann_Rushing-at-baylor.edu
(254) 755-2911





From: Garber, Charles A. :      cgarber-at-2spi.com
Date: Sat, 26 Jul 97 00:48:53 -0500
Subject: Data storage

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

Bob Citron wrote:
==================================================
I agree with your comments that we sometimes try to save "all" of the data
we generate, and much of it is of little future value. But having worked
in the medical device industry for 20 years now, I would like to inject my
two cents, just to provide another viewpoint.
==================================================
Actually, it is not just the medical device industry that has these concerns
. The entire industry of analytical and testing laboratories have these
concerns as well. You see, in our litigious society in America, there is no
such thing as a "statute of limitations" for professional liability exposure
(negligence as some would say). The "clock" starts ticking, not when the
alleged error occurred, but when the alleged error is discovered. This
means, in our case, we have to maintain complete records of all work done
going back to the inception of our business in 1970 with all of the
associated costs.

When these records are not maintained or are not kept available in
retrievable form, then someone who is trying to make it seem like you have
made some error years ago, really is in the driver's seat, that is, they can
say whatever they want to say and you have nothing in your files that says
otherwise. Worse yet, the people who did the work might not remember what
really did go on, say twenty years ago, or can be either retired, no longer
alive or just not anywhere to be found.

And what has to be kept is not only the original data from specific samples,
but also the ancient records showing service history, instrument
calibrations, and other test and measurement procedures that document that
the instrumentation was operating the way it was claimed to have been
operating.

Are there really instances where old archival information of this type is
actually needed? You bet. Do laboratories or individual consultants get
sued? You bet. And how do they fare without such supporting data and
information? Not very well.

Disclaimer: From 9/76-3/97 served as shareholder and director of ILAC Ltd.,
Hamilton, Bermuda, an insurance company that insures analytical and testing
laboratories for professional liability risk.

Chuck


===================================================
Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400
President 1-(800)-2424-SPI
Structure Probe, Inc. FAX: 1-(610)-436-5755
PO BOX 656 e-mail: cgarber-at-2spi.com
West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com


Look for us!
############################
WWW: http://www.2spi.com
############################
==================================================




From: Mary Mager :      mager-at-unixg.ubc.ca
Date: Fri, 25 Jul 1997 21:47:02 -0700
Subject: Re: Hummer VI-a sputter coater

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Dear All,
The supplier of sputter coater targets in Nevada is Abe Dayani of
Refining Systems Inc.
P.O. Box 72466
Las Vegas, NV 89170
phone: 702-368-057
fax: 702-368-0933
He is a buyer of estate jewelry and will make discs of any precious metal or
alloy for much closer to the list price of the metal than most suppliers. I
believe that if you send him a target he will stick the new disc on it.
Woody wrote:
} Any target the correct diameter will work. What I have done is clean-up
} the
} spent target assembly, then silver-epoxy a new (generic) disk to the target
} assembly face. Most EM suppliers sell disks. Material can also be
} purchased
} from suppliers like Alfa Aesar (Johnson Matthey) or Goodfellow. There is a
} supplier in Nevada (I think) who is reputed to be less expensive, but I
} don't
} have any more info available...Maybe someone else knows that name/address.
}
} Woody White
} Mcdermott Technology Inc.
}
} Alt:
} woody.white-at-worldnet.att.net
} http://www.geocities.com/capecanaveral/3722
} ______________________________ Reply Separator
} _________________________________
} Subject: Hummer VI-a sputter coater
} Author: greg-at-umic.sunysb.edu_at_internet at X400post
} Date: 7/24/97 2:59 PM
}
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America





From: dmrelion-at-world.std.com (donald j marshall)
Date: Sat, 26 Jul 1997 08:01:05 -0400
Subject: SLMS

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Fellow microscopy list members

For those of you who are not already familiar with it, I would like
to introduce the Society for Luminescence Microscopy and Spectroscopy
(SLMS). Members of this society are involved in cathodoluminescence (CL) and
UV-excited fluorescence microscopy. Most of the members are involved with
the earth sciences or the material sciences and ceramics. There are also
some applications in forensics, archaeology, and other fields. The society
has been in existence for about 10 years. There is a semi-annual newsletter,
edited by Professor Kopp of the U. Tennessee. Dues are only $10.00 for full
members and $5.00 for students.
The instrumentation used includes the familiar SEMs and EMPAs and a
significant number of the members are using relatively simple and
inexpensive cold cathode based electron beam systems which are small enough
in size that they can be mounted directly on the stage of a conventional
monocular or binocular transmitted light microscope. The CL of the mineral
samples can then be seen directly, in real time, with a minimum of sample
preparation and the information revealed on impurity distributions, etc, is
otherwise very difficult or impossible to obtain with other existing
techniques.

The SLMS has a standards program which has dealt, so far, with the
comparison of photographic results on a complexly zoned carbonate and with
the comparison of the CL emission spectra from a Dy-doped zircon.

For more information on the SLMS, please visit our web site at
http://zephyr.rice.edu/SLMS/SLMS.html This web site is maintained by
Jinny Sisson at Rice University.

If you have any questions regarding the SLMS or the applications of
CL, please feel free to contact me or one of the addresses on the web page.

Disclaimer: I am one of the many charter members of the SLMS and
presently chairperson of the Standards Committee. I have a commercial
interest in furnishing cold cathode based ebeam systems for CL
investigations.

Donald J. Marshall

Donald J. Marshall
Relion Industries
P.O. Box 12
Bedford, MA 01730
Ph: 617-275-4695
FAX: 617-271-0252


email dmrelion-at-world.std.com





From: paul.fischione-at-internetmci.com
Date: Sun, 27 Jul 1997 12:03:32 -0500
Subject: Fwd: Etching with gas

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-- [ From: Paul E. Fischione * EMC.Ver #2.3 ] --

Hello All,

To further comments from Robert H. Olley and Cynthia Bennett regarding
plasma etching, I would like to share a little insight. Ion damage and
"burn marks" are related to the type of plasma generation system. In the
past several years there has been a significant amount of effort expended
to reduce the ion energies present in the plasma. The greatest drive has
been in the semiconductor industry which requires absolute cleaning or
etching without any uncontrolled altering of the material being processed.

Two types of low energy plasma creation have been developed as a result,
namely, inductively coupled and electron cyclotron resonance (ECR) which
create ion energies of 10-15eV and 3-5eV, respectively. At these ion
potentials, insufficient energy exists to either heat or alter the
specimen. Organics are removed from the surface, or material is
selectively removed from the specimen/substrate through the application of
reactive gas species which are generated by the plasma. This process is
purely chemical and does not rely on the forces of ion impingement to
remove material.

To remove organics, oxygen is the ideal process gas which chemically
converts hydrocarbons to CO, CO2 and H2O. To selectively etch materials,
various process gases can be applied. This technology can be readily
utilized without any concern over damage to the specimen. Robert Olley's
application of the atomic oxygen generated in a radio frequency discharge
began to touch on an appropriate application of plasma.

I hope that this clarifies some of the concerns over the use of plasmas.

Regards,

Paul

Paul E. Fischione
E.A. Fischione Instruments, Inc.
9003 Corporate Circle
Export, PA 15632 USA
Phone (412)325-5444
FAX (412)325-5443
Web-site: www.fischione.com




From: SONEJA A K :      soneja-at-giasbma.vsnl.net.in
Date: Mon, 28 Jul 1997 13:36:03 -0500 (GMT)
Subject: uni: Need help on obtaining CITZAF program

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*************************************************************************
Anish Soneja
Director
MENZEL LABORTECHNIK
-Your Imaging Solution Provider
327 Wadala Udyog Bhavan,Wadala,MUMBAI(BOMBAY )400 031.INDIA
Tel:91 22 4145057/4165650
Fax 91 22 4168757
Email:soneja-at-giasbma.vsnl.net.in
*************************************************************************





From: SONEJA A K :      soneja-at-giasbma.vsnl.net.in
Date: Mon, 28 Jul 1997 13:45:50 -0500 (GMT)
Subject: unsubscibe

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unsubscribe

*************************************************************************
Anish Soneja
Director
MENZEL LABORTECHNIK
-Your Imaging Solution Provider
327 Wadala Udyog Bhavan,Wadala,MUMBAI(BOMBAY )400 031.INDIA
Tel:91 22 4145057/4165650
Fax 91 22 4168757
Email:soneja-at-giasbma.vsnl.net.in
*************************************************************************





From: SONEJA A K :      soneja-at-giasbma.vsnl.net.in
Date: Mon, 28 Jul 1997 17:42:25 -0500 (GMT)
Subject: Re: unsubscribe

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unsubscribe

*************************************************************************
Anish Soneja
Director
MENZEL LABORTECHNIK
-Your Imaging Solution Provider
327 Wadala Udyog Bhavan,Wadala,MUMBAI(BOMBAY )400 031.INDIA
Tel:91 22 4145057/4165650
Fax 91 22 4168757
Email:soneja-at-giasbma.vsnl.net.in
*************************************************************************

On Wed, 16 Jul 1997, SONEJA A K wrote:

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} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} unsubscribe
}
} *************************************************************************
} Anish Soneja
} Director
} MENZEL LABORTECHNIK
} -Your Imaging Solution Provider
} 327 Wadala Udyog Bhavan,Wadala,MUMBAI(BOMBAY )400 031.INDIA
} Tel:91 22 4145057/4165650
} Fax 91 22 4168757
} Email:soneja-at-giasbma.vsnl.net.in
} *************************************************************************
}
}





From: Tamara Howard :      howard-at-cshl.org
Date: Mon, 28 Jul 1997 10:50:25 -0400 (EDT)
Subject: non-fluorescing plastic?

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Does anyone have a source for plastic slides/coverslips/dishes that can be
used for tissue culture and won't autofluoresce? Some of the catalogues
list products, but I haven't tried any of them and would like some
feedback from the experts :)

Thanks!

Tamara
CSHL






From: Greg :      greg-at-umic.sunysb.edu
Date: Mon, 28 Jul 1997 11:09:47 -0400
Subject: Anatech LTD new address

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Thanks to all who responded to my Hummer Sputter target question. Many
of you have an adress for Anatech that is old.
The new address is:
Anatech LTD.
6621-F Electronic Dr.
Springfield, Va 22151-4303
1-800-752-7629, Fax 703-941-8077

Greg Rudomen
S.U.N.Y. at Stony Brook
University Microscopy Imaging Center
greg-at-umic.sunysb.edu




From: Chism, Sharron :      SharronChism-at-hmhs.com
Date: Mon, 28 Jul 1997 10:10:00 -0500
Subject: Beam Alignment

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Greetings All,
I'm curious about how often everyone aligns the beam on their 'scopes.
I use a JEOL - JEM 100CX II. In the past, the alignment procedure
was done every morning. Now, we only do it once a week. We don't seem
to have any problems, and the photos are crisp. What are others doing?

Sharron G. Chism HT (ASCP)
Electron Microscopy Lab
Harris Methodist Hospital
Fort Worth, Texas




From: Greg :      greg-at-umic.sunysb.edu
Date: Mon, 28 Jul 1997 12:46:40 -0400
Subject: Re: Beam Alignment

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Chism, Sharron wrote:

} I'm curious about how often everyone aligns the beam on their 'scopes.
} I use a JEOL - JEM 100CX II. In the past, the alignment procedure
} was done every morning. Now, we only do it once a week. We don't seem
} to have any problems, and the photos are crisp. What are others doing?
}

I check alignment every morning. Hopefully any other problems will
also show up.

Greg-at-unic.sunysb.edu
University Microscopy Imkaging Center
S.U.N.Y. Stony Brook




From: Scott D. Walck WL/MLBT :      walcksd-at-ml.wpafb.af.mil
Date: Mon, 28 Jul 97 13:12:14 -0400
Subject: Re: Beam Alignment

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} ------------------------------------------------------------------------ The
Microscopy ListServer -- Sponsor: The Microscopy Society of America To
Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
-----------------------------------------------------------------------.
}
} Greetings All,
} I'm curious about how often everyone aligns the beam on their 'scopes.
I use a JEOL - JEM 100CX II. In the past, the alignment procedure was
done every morning. Now, we only do it once a week. We don't seem to have
any problems, and the photos are crisp. What are others doing?
}
} Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist
Hospital Fort Worth, Texas

If you have a multi-user environment with users of different experience
levels, every user should perform the quick alignment procedures. When there
is trouble and the machine can't be aligned with the simple procedures, then
the manager or service engineer of the TEM should perform a complete one
(which includes the mechanical alignment). In addition, the microscope should
be left in a standard startup condition for the next user. (Aperture out, low
mag, beam spread to uniformly light the screen, text on negative info screen
erased, and the plate number and info changed to some default, e.g. 0001.)




From: ldb94001-at-uconnvm.uconn.edu (Lisa Brown)
Date: Mon, 28 Jul 1997 13:48:04 +0100
Subject: LM- immunolabelling of frozen muscle sections.....

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I have recently used 10 micron frozen sections of muscle fibers for
immunolabelling using an antibody produced in our lab that is specific for
a protein on the myosin filaments. Visualization was achieved by a
flourescent secondary antibody. The problem is that we are not able to
obtain the resolution necessary with the flourescent antibody in order to
access fiber to fiber variation in labelling within the sarcomere or
between sarcomeres.

I am thinking of using a biotinylated secondary antibody such that a
biotin/avidin horseradish peroxidase visualization system can be used. This
would allow us to continue this study at the light microscope level.
Eventually EM wil be utilized, but light microscopy will allow for broader
asessment of this large muscle. Does anyone know of a dehydration and
counterstaining protocol that can be used for final viewing of the muscle
tissue?






From: Ron Bartley :      ron-at-imii.com
Date: Mon, 28 Jul 1997 14:03:38 -0400
Subject: Flourescence Microscopy/Medical

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I am looking for a list of applications using Flourescence Microscopy.
These can be medical and/or non medical.

If you have such a list or know where I may acquire this list I would
appreciate this information.




From: Escuela Normal :      enrosn-at-satlink.com
Date: Mon, 28 Jul 1997 15:52:25 -0300
Subject: Flourescence Microscopy/Medical

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subscribe microscopy enrosn-at-satlink.com





From: H. Birnbaum :      birnbaum-at-uimrl7.mrl.uiuc.edu
Date: Mon, 28 Jul 1997 14:57:19 -0600
Subject: Replacement for Alwyn Eades

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Dear Colleagues:

As some of you may know, Alwyn Eades is leaving the position as Director of
the Center for Microanalysis of Materials in the MRL for a faculty position
at Lehigh University. We are very sorry to lose Alwyn, but he would like
to affect a career change at this time and we wish him the best of luck.

We will be instituting a search for a new Director of the CMM. At this
time, I am writing to you to inform you of this search and to ask that you
notify any colleagues you feel might be qualified to apply. A description
of the position is attached below. Please feel free to distribute it.

Alwyn has been an excellent Director of the CMM and to replace him we will
need all the help we can get. I do hope you can help us.

Thank you in advance.

Howard Birnbaum

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

PRINCIPAL RESEARCH SCIENTIST
(DIRECTOR OF THE CENTER FOR MICROANALYSIS OF MATERIALS)
FREDERICK SEITZ MATERIALS RESEARCH LABORATORY
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN

The Frederick Seitz Materials Research Laboratory (MRL) at the University
of Illinois at Urbana-Champaign is seeking a Director for its Center for
Microanalysis of Materials (CMM). The CMM is a major analytic
instrumentation center encompassing the techniques of electron microscopy
(TEM, STEM, LEEM and SEM), microchemical and surface analysis (SIMS,
Scanning Auger, XPS, UPS), ion beam methods (RBS, channeling, PIXE), probe
microscopies (STM and AFM), and x-ray methods. It functions to support the
research activities of faculty, Research Associates and students at the
University of Illinois and at other universities, and for researchers at
Federal Laboratories and in industry. The present staff of the CMM
consists of thirteen professional analysts who are expert in the various
analytic methods.

The Director of the CMM reports to the Director of the MRL and the CMM is
supported by the MRL as one of a number of instrumentation centers. The
CMM Director works with the Director of the MRL in planning the development
of the CMM, in the development of new analytic methods, in the purchase of
new and replacement instruments, and in making the CMM increasingly
important in the research endeavor of the MRL. He/she is responsible for
the management of the CMM staff, for providing scientific and technical
expertise to the CMM, and with the CMM staff members, to the users of the
CMM. The Director coordinates staff development and outreach activities to
researchers at the University of Illinois and nationwide. She/he
represents the MRL at appropriate national activities involving
instrumentation facilities. The person selected for the position of
Director is expected to carry out an appropriate research program (for
which support is provided) and to encourage appropriate research activities
on the part of the CMM staff.

Candidates should have a Ph.D. in Physics, Chemistry or Materials Science
and have an established research record in an appropriate field involving
the use of modern analytical methods. The candidates should have strong
technical expertise in at least one of the areas of analysis covered by the
CMM and should be capable of providing technical leadership in all of the
analytic areas. He/she should have an established record of management of
scientific personnel and the ability to develop and manage budgets.

The salary for this position will be commensurate with the experience of
the candidate chosen and full benefits of the University of Illinois will
apply. Please submit applications to: Howard Birnbaum, Director; c/o Ms
Donna Jacobs; Frederick Seitz Materials Research Laboratory; University of
Illinois at Urbana-Champaign; 104 South Goodwin Avenue; Urbana, IL 61801.
The application material should include a resume', a statement of your
views on the operation of a microanalysis facility within a large
university research establishment such as the MRL, the names and contact
information for five references who are familiar with aspects of your
career important to your qualifications for the position. Applications
received by November 1, 1997 will be fully considered but acceptance of
applications and screening of applicants will continue until the position
is filled.

The University of Illinois is an equal opportunity / affirmative action
employer,

Howard K. Birnbaum (217)
333-1370
Materials Research Laboratory FAX (217) 244-2278
University of Illinois e mail: birnbaum-at-uimrl7.mrl.uiuc.edu
Urbana, IL 61801






From: William Tivol :      tivol-at-wadsworth.org
Date: Mon, 28 Jul 1997 16:07:45 -0500 (EDT)
Subject: Re: Beam Alignment

Contents Retrieved from Microscopy Listserver Archives
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} I'm curious about how often everyone aligns the beam on their 'scopes.
} I use a JEOL - JEM 100CX II. In the past, the alignment procedure
} was done every morning. Now, we only do it once a week. We don't seem
} to have any problems, and the photos are crisp. What are others doing?
}
Dear Sharron,
We go through an extensive procedure daily. In addition for dif-
fraction we also check the condenser aperture position and do further align-
ment in selected area and diffraction modes.
About once a month we check the mechanical alignment of the objective
upper pole piece, and once a year we disassemble and clean the entire lens
column and do a complete mechanical alignment.
I don't think the normal user--doing imaging of thick biological
specimens at ~10k mag--would notice the difference in pictures if we didn't
do the alignment as often, but there are uses for which it is critical, and
it tells us a lot about the machine. Since we do all the maintenance--no
service contract for the HVEM--the information we get from doing frequent
alignments is quite useful.
Yours,
Bill Tivol




From: William Tivol :      tivol-at-wadsworth.org
Date: Mon, 28 Jul 1997 16:07:45 -0500 (EDT)
Subject: Re: Beam Alignment

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

} I'm curious about how often everyone aligns the beam on their 'scopes.
} I use a JEOL - JEM 100CX II. In the past, the alignment procedure
} was done every morning. Now, we only do it once a week. We don't seem
} to have any problems, and the photos are crisp. What are others doing?
}
Dear Sharron,
We go through an extensive procedure daily. In addition for dif-
fraction we also check the condenser aperture position and do further align-
ment in selected area and diffraction modes.
About once a month we check the mechanical alignment of the objective
upper pole piece, and once a year we disassemble and clean the entire lens
column and do a complete mechanical alignment.
I don't think the normal user--doing imaging of thick biological
specimens at ~10k mag--would notice the difference in pictures if we didn't
do the alignment as often, but there are uses for which it is critical, and
it tells us a lot about the machine. Since we do all the maintenance--no
service contract for the HVEM--the information we get from doing frequent
alignments is quite useful.
Yours,
Bill Tivol




From: Greg :      greg-at-umic.sunysb.edu
Date: Mon, 28 Jul 1997 16:47:01 -0400
Subject: Re: Anatech LTD new address

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Message-ID: {33DCB63B.1CE825AB-at-umic.sunysb.edu}

Greg wrote:
}
} Thanks to all who responded to my Hummer Sputter target question. Many
} of you have an adress for Anatech that is old.
} The new address is:
} Anatech LTD.
} 6621-F Electronic Dr.
} Springfield, Va 22151-4303
} 1-800-752-7629, Fax 703-941-8077
}
} Greg Rudomen
} S.U.N.Y. at Stony Brook
} University Microscopy Imaging Center
} greg-at-umic.sunysb.edu




From: Chism; Sharron :      SharronChism-at-hmhs.com at -SMTPLink
Date: 7/28/97 10:10 AM
Subject: Beam Alignment

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Sharron,

I generally do at least some cursory alignment procedures each time I sit at the
microscope. Most of the time all is well, but sometimes the last person had it
in a different mode and/or did not bring it back to a "standard state".
However, like stretching before excercising, I also find that running through
the alignment procedures gets me in the proper state of mind for doing
microscopy (as well as allowing my eyes to get dark-adjusted). Therefore, I
would run through it just for that purpose. Is that kind of a Zen thing? :-)

Cheers,

John Vetrano
_______________________________________________________________________________

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

Greetings All,
I'm curious about how often everyone aligns the beam on their 'scopes.
I use a JEOL - JEM 100CX II. In the past, the alignment procedure
was done every morning. Now, we only do it once a week. We don't seem
to have any problems, and the photos are crisp. What are others doing?

Sharron G. Chism HT (ASCP)
Electron Microscopy Lab
Harris Methodist Hospital
Fort Worth, Texas




From: Steve Chapman :      PROTRAIN-at-CompuServe.COM
Date: Mon, 28 Jul 1997 18:05:51 -0400
Subject: Beam Alignment

Contents Retrieved from Microscopy Listserver Archives
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I started life as an EM service engineer in 1966, since then I think I ha=
ve
handled almost every type of commercial TEM and SEM, therefore I have bee=
n
able to take a good look at the subject of alignment.

How often you need to align a TEM depends very much on its stability, =

thermal and mechanical. If you leave the lenses switched on at a
controlled temperature, about 2 deg centigrade below room temperature is
ideal, there is no reason for constantly aligning the lenses. =


If the machine has mechanical lens alignment it is my experience that the=

more you move them the more they move, compromise! However you must do a=

quick check prior to use of 1. gun alignment - viewing the spot and halo=
:
2. illumination - in relation to the screen centre: 3. condenser apertur=
e
- in relation to the screen when C2 is spread overfocus: 4. objective
aperture - in relation to the diffraction spot.

Of course the alignments that you need to perform should relate to the
levels that you expect to reach when using the instrument. Less than
10,000X then you can get away with a very quick check of saturation. =

Should your target be 500,000X then the most important alignment will be
voltage alignment, as clearly resolution will be your goal. In this case=

forget using the instrument within 2 hours of switching on the high
voltage; that is unless you have a gas filled HT tank. It takes this tim=
e
for the high voltage to stabilise due to heat gained in the tank being
required to reach the same level as heat lost. Gas filled tanks seem to
take about 45 minutes at 120kV!

Most people over align their instruments as if the feat of completing the=

alignment is part of their religion! =

The time when an alignment becomes critical is when the lens in question =
or
the high voltage become unstable. To mis align the lenses is a standard
engineer trick to isolate a fault to a particular part of the instrument.=
=

See my book "Maintaining & Monitoring the Transmission electron Microscop=
e"
published by the Royal Microscopical Society ISBN 0-19-856407-4. This bo=
ok
also outlines all the TEM alignment procedures, including deflection coil=
s
and stigmators as well as covering typical image defects due to alignment=

and instability problems.

In spite of what some manufacturers may claim the alignment of a TEM
follows basic procedures which have not changed since the 1960s, how can
they, they are just electron optics!

Steve Chapman
Senior Consultant
Protrain




From: Henrik Kaker :      Henrik.Kaker-at-guest.arnes.si
Date: Thu, 28 Aug 1997 00:16:53 +0200
Subject: Re: Anatech LTD new address

Contents Retrieved from Microscopy Listserver Archives
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Greg wrote:
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} Thanks to all who responded to my Hummer Sputter target question. Many
} of you have an adress for Anatech that is old.
} The new address is:
} Anatech LTD.
} 6621-F Electronic Dr.
} Springfield, Va 22151-4303
} 1-800-752-7629, Fax 703-941-8077
}
} Greg Rudomen
} S.U.N.Y. at Stony Brook
} University Microscopy Imaging Center
} greg-at-umic.sunysb.edu

Web address of Anatech LTD is:

http://www.anatechltd.com

--
Henrik Kaker
SEM-EDS Laboratory, Metal Ravne d.o.o.
Koroska c.14, 2390 Ravne, Slovenia
Tel: +386-602-21-131, Fax: +386-602-20-436
SEM-EDS Laboratory Web Site
http://www2.arnes.si/guest/sgszmera1/index.html
Microscopy Vendors Database
http://www.kaker.com/mvd/vendors.html
Kaker.Com
http://www.kaker.com




From: Steve Collins :      stevesbt-at-erols.com
Date: Mon, 28 Jul 1997 18:29:26 -0400
Subject: Etching with Gas

Contents Retrieved from Microscopy Listserver Archives
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Hello All:

In reference to Cynthia Bennett's and Robert H. Olley's information request,
there are in fact commercially available multi-process radio frequency etching
systems at reasonable costs. For a full range of etch capability, it is
necessary to have multi-gas inputs for user determined ratios of more than
one species and the versatility to select more than one etching gas. Various
gases can be used for selective etch of specific materials as has been done in
the semiconductor industry since the 1960's. There are a couple reference
texts available with specific designs of etching systems including parallel
plate, ECR, inductivly coupled plasma and microwave technology.
"Thin Film Processes" by Vossen, version I (1978) and II is an excellent
reference to these processes. Also "Glow Discharge Processes" by Chapman
(1980) covers these techniques in detail. As indicated in other responses,
oxygen is most common for etching or removing hydrocarbons, organics and
polymers such as photoresist. The O2 gas provides a chemical etch and in
some cases it is desired to speed up the process by adding a heavier,
non-reactive atom such as argon. This will cause a physical etch as well
as a reactive chemical etch process.

To avoid "advertising" on this network, please refer to the South Bay
Technology home page for additional information on one commercial
versatile RF plasma etching system or contact me direct for details
on the unit.

Regards,
Steve

Steve Collins
scollins-at-southbaytech.com
Ph: 703-486-7999 (east coast USA)
714-492-2600 (west coast USA)
800-728-2233 (Toll Free)
Fax: 714-492-1499
Web Page: http://www.southbaytech.com




From: Melvyn Dickson :      M.Dickson-at-unsw.edu.au
Date: Tue, 29 Jul 1997 10:12:16 +1000
Subject: Re: Beam Alignment

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At 10:10 AM 7/28/97 -0500, you wrote:
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America

Column alignment of TEMs is mostly for convenience and does not much affect
resolution.

Filament traverse and tilt are so a maximal amount of beam enters the
condenser system

Condenser alignment is so the illumination stays on the screen when
condenser lens focal length is changed. The condenser/gun system is
aligned with the axis of the objective so the illumination stays on the
screen as objective focal length is changed.

Imaging lenses have their axes put on the line joining the centre of the
objective with the centre of the screen so the image stays central and in
view when magnification is altered.

The objective aperture MUST be centred on the axis of the objective. We
use the zero order spot in the diffration pattern to define the axis.

For best resolution the entire illumination system should be tilted so its
axis coincides with either the current or voltage centre of the objective.
Voltage centre has an advantage as it aligns with the mean axes of all the
imaging lenses, not just the objective. But there is not much in it. So
when you are shooting for really good resolution, centre the objective
aperture, correct astigmatism using background phase speckle on your
specimen, use high voltage modulation to test illumination tilt and adjust
for minimal image wobble, and shoot.

Maybe once a week I check condenser aperture alignment, every operator must
check objective aperture alignment, gun alignment is checked after filament
change of voltage change. The rest doesn't matter. We have not done a
full alignment on our Hitachi H-7000 in 7 years.
Mel Dickson
President, Australian Society for Electron Microscopy
Director, Electron Microscope Unit,
University of New South Wales.
Sydney NSW 2052 Australia

Phone (+612) 9385-2945
Fax (+612) 9385-1067

Website {http://www.unsw.edu.au/clients/emu_top.htm}




From: :      MAILER-DAEMON-at-med.vu.nl
Date: Tue, 29 Jul 97 3:52:58 +0200
Subject: Etching with Gas

Contents Retrieved from Microscopy Listserver Archives
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Received: from mx05.erols.com (mx05.erols.com [205.252.116.249]) by mail0.erols.com (8.8.5/8.7.3/970701.001epv) with ESMTP id WAA20834 for {stevesbt-at-mail0.erols.com} ; Mon, 28 Jul 1997 22:01:06 -0400 (EDT)
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for {stevesbt-at-erols.com} ; Mon, 28 Jul 1997 21:55:21 -0400
Received: by asklepios.med.vu.nl with VINES-ISMTP; Tue, 29 Jul 97 3:52:51 +0200

Hello All:

In reference to Cynthia Bennett's and Robert H. Olley's information request,
there are in fact commercially available multi-process radio frequency etching
systems at reasonable costs. For a full range of etch capability, it is
necessary to have multi-gas inputs for user determined ratios of more than
one species and the versatility to select more than one etching gas. Various
gases can be used for selective etch of specific materials as has been done in
the semiconductor industry since the 1960's. There are a couple reference
texts available with specific designs of etching systems including parallel
plate, ECR, inductivly coupled plasma and microwave technology.
"Thin Film Processes" by Vossen, version I (1978) and II is an excellent
reference to these processes. Also "Glow Discharge Processes" by Chapman
(1980) covers these techniques in detail. As indicated in other responses,
oxygen is most common for etching or removing hydrocarbons, organics and
polymers such as photoresist. The O2 gas provides a chemical etch and in
some cases it is desired to speed up the process by adding a heavier,
non-reactive atom such as argon. This will cause a physical etch as well
as a reactive chemical etch process.

To avoid "advertising" on this network, please refer to the South Bay
Technology home page for additional information on one commercial
versatile RF plasma etching system or contact me direct for details
on the unit.

Regards,
Steve

Steve Collins
scollins-at-southbaytech.com
Ph: 703-486-7999 (east coast USA)
714-492-2600 (west coast USA)
800-728-2233 (Toll Free)
Fax: 714-492-1499
Web Page: http://www.southbaytech.com







From: rgarcia-at-nova.wright.edu
Date: Tue, 29 Jul 1997 08:35:19 -0500 (EST)
Subject: Re: Beam Alignment

Contents Retrieved from Microscopy Listserver Archives
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Sharron,

I also have a 100CX. I align the microscope everytime I sit down
to it. If it is well aligned already the whole time takes just a few
minutes out of my day. It's just a good habit to get into particularly if
you have a multiuser environment.




From: John G. Aghajanian, Ph.D. :      johna-at-scientist.com
Date: Tue, 29 Jul 1997 09:18:36 -0400
Subject: Epon 812

Contents Retrieved from Microscopy Listserver Archives
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G'Day TEM-types,

And please pardon the bandwidth if anybody feels it's inappropriate. =
This is the easiest way to widely disperse the message.

I have a substantial supply of Epon 812 (the real thing) that I find I =
must part with. If anyone is interested in retro-embedding their =
specimens, please contact me privately.

johna-at-scientist.com

John G. Aghajanian, Ph.D.





From: Robert Underwood :      underwoo-at-u.washington.edu
Date: Tue, 29 Jul 1997 07:30:37 -0700 (PDT)
Subject: Re: LM- immunolabelling of frozen muscle sections.....

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Hi Lisa,

If you have access to a microscope with DIC (differential interference
contrast) You can visualize the muscle very nicely and see the peroxidase
product. You can also counterstain with hematoxylin in get more reference
points.

Bob
Morphology Core

On Mon, 28 Jul 1997, Lisa Brown wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
} I have recently used 10 micron frozen sections of muscle fibers for
} immunolabelling using an antibody produced in our lab that is specific for
} a protein on the myosin filaments. Visualization was achieved by a
} flourescent secondary antibody. The problem is that we are not able to
} obtain the resolution necessary with the flourescent antibody in order to
} access fiber to fiber variation in labelling within the sarcomere or
} between sarcomeres.
}
} I am thinking of using a biotinylated secondary antibody such that a
} biotin/avidin horseradish peroxidase visualization system can be used. This
} would allow us to continue this study at the light microscope level.
} Eventually EM wil be utilized, but light microscopy will allow for broader
} asessment of this large muscle. Does anyone know of a dehydration and
} counterstaining protocol that can be used for final viewing of the muscle
} tissue?
}
}
}





From: m-moody-at-nwu.edu (Maya Moody)
Date: Tue, 29 Jul 1997 09:44:58 -0500
Subject: Fujix Pictography 3000

Contents Retrieved from Microscopy Listserver Archives
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Hi,

Just wondering where people are getting supplies
for their Fujix Pictography 3000 printers.

Thanks,


Maya Moody
Northwestern University
Cell Imaging Facility
312-503-4445
m-moody-at-nwu.edu





From: Simon C. Watkins :      swatkins-at-pitt.edu
Date: Tue, 29 Jul 1997 10:48:25 -0400
Subject: cost of supplies for Pictrography

Contents Retrieved from Microscopy Listserver Archives
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Hi folks
We are trying to find the most economic source for FUji Pictrography
supplies, I would welcome info on where current users buy their
materials and how much they pay for the various components. I will
compile the results and send back to anyone interested.
If vendors wish to add to this cost survey please contact me directly
Tx


--
Simon C. Watkins Ph.D.
Associate Professor
Director CBI
University of Pittsburgh
Pittsburgh PA 15261
tel:412-648-3051
Fax:412-648-2004
URL:http://sbic6.sbic.pitt.edu






From: HILDEGARD CROWLEY :      hcrowley-at-du.edu
Date: Tue, 29 Jul 1997 11:02:50 -0600 (MDT)
Subject: LM:Mess-Polyclonal-mono-control

Contents Retrieved from Microscopy Listserver Archives
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Dear Immuno LM Friends,

We are in a tight spot. We have collected data with a monoclonal
antibody (Snap-25, reactive with fusion proteins from COS cells - the
immunogen was crude synaptic immunoprecipitate from humans) which was
raised in mouse. We used rats for data
collection. Our secondary was FITC-conjugated AFfiniPure Goat Anti-mouse
IgG. Now we find that we cannot obtain a peptide for control purposes.
And, if we omit the primary, and use only the above secondary, we get a
definite immuno response: It looks exactly as though the primary had been
applied. The manufacturer of the FITC
states that the anti-mouse antibody may cross-react with other species.
Now What?
We cannot purchase polyclonal Snap-25. Does anyone have any ideas? Did
we make a huge mistake in not testing for control situations at the outset?
We are relatively new at this game - do others get down these blind
alleys, singing and dancing all the way until the lights go out?
Bye,
Hildy




From: Chism, Sharron :      SharronChism-at-hmhs.com
Date: Tue, 29 Jul 1997 12:33:00 -0500
Subject: Re: Beam Alignment ... Thank You!

Contents Retrieved from Microscopy Listserver Archives
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Thanks to all who responded to my question on the frequency of beam
alignment. It was interesting to hear from all of you. My favorite
response included the advice of "If it ain't broke, don't fix it!". The
majority of those that answered do a quick alignment every day ...
mostly because of so many pairs of hands that fiddle with the 'scope
during the day. Since I am the only tech that works with this 'scope,
and always return it to "square one", and only have two pathologists
that operate it ... once a week alignment seems to be all it needs.
Our normal operation takes us up to 14k or 20k mag and seldom higher
than 60k - 80k. (The specimens are about 70nm in thickness.) Still
others have said that they, too, have a CX100 and only align the beam
once a week. The information has been very helpful. Thanks, again for
your info!

Sharron G. Chism HT (ASCP)
Electron Microscopy Lab
Harris Methodist Hospital
Fort Worth, Texas




From: Robert Kayton :      kayton-at-ohsu.edu
Date: Tue, 29 Jul 1997 11:28:52 -0700
Subject: Microscopy & Microanalysis

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Hello from Portland site for M & M 1999,

Our Local Arrangements Committee is chewing over a question that on which we
have differing opinions. The question is " What is the true function of the
Sunday social and how does this function relate to the venue?" We have come
up with a couple of options.

Option 1: Is this function serving to rekindle acquaintances between
colleagues.

Option 2: Is this function serving to not only rekindle acquaintances
between colleagues but also to set the tone for the coming meeting, and show
case the hosting city.


For those list members that attend the meetings, and any others that would like
to comment all your input will be welcome. If you want to reply directly to
me, I will be delighted to post a summary after comments have been received.



Bob Kayton, Ph.D.
Histology/E.M. Core Director
C.R.O.E.T.
Oregon Health Sciences University
Portland, OR 97201
W-503-494-2504
Fax-503-494-6831
H-503-590-7801




From: Linda Barthel :      barthel-at-umich.edu
Date: Tue, 29 Jul 1997 16:09:37 -0400 (EDT)
Subject: Re: LM:Mess-Polyclonal-mono-control

Contents Retrieved from Microscopy Listserver Archives
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Jackson Immunoresearch carries secondary antibodies that are pre-absorbed
against several different animal seras including rat. We routinely do
double label ICC with mouse and rat monoclonals and purchase our secodary
antibodies from them. See Braisted et al. Development 120:2409-2419
(1994). There is no cross reactivity seen with the mouse secondary to the
rat immunoglobins. These preabsorbed antibodies should work well on your
mouse tissue. Other companies carry pre-absorbed secondaries also. We
have been using Jackson for years and have been very satisfied with them.
Linda Barthel
Research Associate II
Department of Anatomy and Cell Biology
University of Michigan
lab (313) 764-7476
fax (313) 763-1166
barthel-at-umich.edu



On Tue, 29 Jul 1997, HILDEGARD CROWLEY wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
}
} Dear Immuno LM Friends,
}
} We are in a tight spot. We have collected data with a monoclonal
} antibody (Snap-25, reactive with fusion proteins from COS cells - the
} immunogen was crude synaptic immunoprecipitate from humans) which was
} raised in mouse. We used rats for data
} collection. Our secondary was FITC-conjugated AFfiniPure Goat Anti-mouse
} IgG. Now we find that we cannot obtain a peptide for control purposes.
} And, if we omit the primary, and use only the above secondary, we get a
} definite immuno response: It looks exactly as though the primary had been
} applied. The manufacturer of the FITC
} states that the anti-mouse antibody may cross-react with other species.
} Now What?
} We cannot purchase polyclonal Snap-25. Does anyone have any ideas? Did
} we make a huge mistake in not testing for control situations at the outset?
} We are relatively new at this game - do others get down these blind
} alleys, singing and dancing all the way until the lights go out?
} Bye,
} Hildy
}





From: Jacky Larnould :      larnould-at-mnet.fr
Date: Tue, 29 Jul 1997 22:48:21 +0200
Subject: Beam alignment

Contents Retrieved from Microscopy Listserver Archives
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Hi all

In my opinion concerning 100C(X) alignment, don't make knots to your neurons!
100 cx is a very complete but simple microscope to use.Only 6 lenses and 4
alignments coils
including condenser and objective stigmators.
All I'm describing is for routine work, for High resolution job it's an
other problem.
The main mechanical alignment is intermediate and projector lens and must
be perform once or two times a
year. alignment of condenser aperture is OK until people change the
aperture. Gun tilt and shift are generally OK
until you change the filament.
The only thing to check is Spot size 1 and center with Gun shift then spot
size 3 and center with Trans and repeat
(once or two time) until the position of beam is the same between spot 1
and 3. If you have turn a lot the gun shift, you can check the maximunm of
brightness or the image filament with gun tilt.
If the precedent user has made dark field you can check the voltage center
with HV wobbler and tilt knob.
That's all, no more than a couple of minutes. Of course centering of
objective aperture and setting of objective stig must be perform by user
but it's not alignment it's just like to focus the image to obtain the best
you can have.
Hope that helps.
==========================================================
Jacky Larnould
mailto:larnould-at-mnet.fr
voice:33 (0)4 67 72 28 26
fax :33 (0)4 67 79 54 90




From: James Martin :      James.S.Martin-at-williams.edu
Date: Tue, 29 Jul 1997 16:50:25 -0400 (EDT)
Subject: ISO 9001

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I am looking for information about the ISO 9001 standard with regard to
the operation of FT-IR microscopy and SEM systems for materials analysis,
and would very much appreciate communicating with list members who have
practical experience in this area.

Many thanks in advance.

James Martin
Williamstown Art Conservation Center







From: Julian Smith III :      smithj-at-Winthrop.edu
Date: Tue, 29 Jul 1997 17:45:26 -0400
Subject: EM400 tem/stem: free to non-profit

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Hi all:
EMSL has a Philips 400 TEM/STEM to give away to any qualified non-profit.
The instrument is in good condition and is located in Greensboro, NC.
Recipient is responsible for packing and moving the instrument.
Interested parties should contact Ron Mahoney at EMSL 910-297-1487 for more
information.
Cheers,
Julian

Julian P.S. Smith III
Biology
Winthrop University
Rock Hill, SC 29733
803-323-2111 x227 (vox)
803-323-2246 (fax)






From: Sara Miller :      saram-at-acpub.duke.edu
Date: Tue, 29 Jul 1997 18:19:13 -0400 (EDT)
Subject: Re: LM:Mess-Polyclonal-mono-control

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On Tue, 29 Jul 1997, HILDEGARD CROWLEY wrote:

} Date: Tue, 29 Jul 1997 11:02:50 -0600 (MDT)
} From: HILDEGARD CROWLEY {hcrowley-at-du.edu}
} To: postmessage {Microscopy-at-sparc5.microscopy.com}
} Subject: LM:Mess-Polyclonal-mono-control
}
} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
}
} Dear Immuno LM Friends,
}
} We are in a tight spot. We have collected data with a monoclonal
} antibody (Snap-25, reactive with fusion proteins from COS cells - the
} immunogen was crude synaptic immunoprecipitate from humans) which was
} raised in mouse. We used rats for data
} collection. Our secondary was FITC-conjugated AFfiniPure Goat Anti-mouse
} IgG. Now we find that we cannot obtain a peptide for control purposes.
} And, if we omit the primary, and use only the above secondary, we get a
} definite immuno response: It looks exactly as though the primary had been
} applied. The manufacturer of the FITC
} states that the anti-mouse antibody may cross-react with other species.
} Now What?
} We cannot purchase polyclonal Snap-25. Does anyone have any ideas? Did
} we make a huge mistake in not testing for control situations at the outset?
}
YES

We are relatively new at this game - do others get down these blind
} alleys, singing and dancing all the way until the lights go out?

SUGGEST YOU TALK TO AN IMMUNOLOGIST TO HELP DESIGN YOUR IMMUNOEXPERIMENTS.
THERE ARE LOTS OF REACTIVITIES THAT EVEN EXEPRIENCED MICROSCOPISTS DON'T
KNOW ABOUT WHICH
ARE COMMON KNOWLEDGE AMONG IMMUNOLOGY FOLKS. FORTUNATELY, I MARRIED
AN IMMUNOPATHOLOGIST WITH A PH D IN IMMUNOLOGY--HE SOLVES MY REACTIVITY
PROBLEMS.

} Bye,
} Hildy
}
Rats and mice are very similar. I'm not surprised you got cross
reactivity. You should always do negative controls-- in every
experiment--one of which is your secondary without the primary. But this
is not enough; you should have a primary control (one that is the same
type as your experimental), either a preimmune serum if polyclonal or a
non-reactive mono (proven non-reactive) that is the same species and same
isotype.

You can buy a rat antimouse-FITC which probably won't react with rat tissue
(ours didn't). Also, you can try absorbing your secondary with normal mouse
serum, but you will probably lose a lot of your specific reactivity too.

Good luck,
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: Nestor J. Zaluzec :      zaluzec-at-Sparc5.Microscopy.Com
Date: Tue, 29 Jul 1997 18:14:10 -0500
Subject: Ask-A-Microscopist

Contents Retrieved from Microscopy Listserver Archives
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Anyone have any suggestions for the question below. This is out
of my area...

Nestor



} Date: Tue, 29 Jul 1997 10:48:01 -0500
} To: Zaluzec-at-sparc5.microscopy.com
} From: lmtuhela-at-cc.owu.edu ()
} Subject: Ask-A-Microscopist
}
} Below is the result of your feedback form. It was submitted by
} (lmtuhela-at-cc.owu.edu) on Tuesday, July 29, 1997 at 10:48:00
} ---------------------------------------------------------------------------
}
} Email: lmtuhela-at-cc.owu.edu
} Name: Laura Tuhela-Reuning
}
} School: Ohio Wesleyan University
}
} State: OH
}
} Zip: 43015
}
} Question: A faculty member is interested in using our SEM to observe the
} giant chromosome in Drosophila for an upcoming genetics class. Are there
} any suggestions for preparing the samples? We have a cryo system
} available as well as variable pressure but do not have a critical point
} dryer. Thank you!
}
} ---------------------------------------------------------------------------
}






From: Henry Lippard :      hlippard-at-charlotte.infi.net
Date: Tue, 29 Jul 1997 20:34:09 -0400
Subject: suscribe digest

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-----------------------
Henry Lippard
hlippard-at-charlotte.infi.net






From: NAGY, Peter :      p.nagy-at-r1.atki.kfki.hu
Date: Wed, 30 Jul 1997 09:06:45 -0700
Subject: subscribe.digest

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From: GREG VAUGHN MARTIN :      gmartin-at-welchlink.welch.jhu.edu
Date: Wed, 30 Jul 1997 08:20:15 -0400 (EDT)
Subject: Re: LM:Mess-Polyclonal-mono-control

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Hey Hildy and other folks --

We do a lot of immunocytochem on Rat tissue and Rat -derived
cultured cells and have (almost) always had good reults using anti-mouse
secondaries from Jackson Immunoresearch (I've no stake in this company).
They have antibodies raised in Donkey that are already preadsorbed
against a number of other species, including Rat. Generally they give
very low non-specific binding on our specimens.
Negative controls are very important. We use the "no
primary" and "pre-immune (non-immune)" controls often, but prefer the
"irrelevent primary" control where possible.

Greg Martin
Dept. of Cell Biology and Anatomy
Johns Hopkins School of Medicine


On Tue, 29 Jul 1997, HILDEGARD CROWLEY wrote:

} ------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
} -----------------------------------------------------------------------.
}
}
} Dear Immuno LM Friends,
}
} We are in a tight spot. We have collected data with a monoclonal
} antibody (Snap-25, reactive with fusion proteins from COS cells - the
} immunogen was crude synaptic immunoprecipitate from humans) which was
} raised in mouse. We used rats for data
} collection. Our secondary was FITC-conjugated AFfiniPure Goat Anti-mouse
} IgG. Now we find that we cannot obtain a peptide for control purposes.
} And, if we omit the primary, and use only the above secondary, we get a
} definite immuno response: It looks exactly as though the primary had been
} applied. The manufacturer of the FITC
} states that the anti-mouse antibody may cross-react with other species.
} Now What?
} We cannot purchase polyclonal Snap-25. Does anyone have any ideas? Did
} we make a huge mistake in not testing for control situations at the outset?
} We are relatively new at this game - do others get down these blind
} alleys, singing and dancing all the way until the lights go out?
} Bye,
} Hildy
}




From: Nancy Smythe :      SMYTHEN-at-smtpgw2.musc.edu
Date: Wed, 30 Jul 1997 08:31:48 -0400
Subject: subscribe

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subscribe
smythen-at-musc.edu

Nancy Smythe





From: Roger Cargill :      Roger.Cargill-at-USDWP.MSU.EDU
Date: Wed, 30 Jul 1997 10:06:18 -0500
Subject: electron microscope available

Contents Retrieved from Microscopy Listserver Archives
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Michigan State University has an electron microscope available it is a Philips model
EM201, purchased in 1972. Anyone interested contact Roger Cargill (517)355-0364
or cargill-at-pilot.msu.edu.
thank you




From: kelloes-at-emlab.cb.uga.edu
Date: Tue, 29 Jul 1997 21:35:44 +0000
Subject: Used microscope,photomicroscope, ultramicrotome

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Hello to all:

We have the following equipment, if anyone is interested:

1. A Vickers Patholux microscope (10x, 40x,75x lenses and condensors
included).

2. B & L Grating monochromator and UV Photomicroscope.

3. OM-U2 ultramicrotome

If anyone is interested, just e-mail me with an offer.
Cathy Kelloes




From: Michael K. Cinibulk :      cinibumk-at-ml.wpafb.af.mil
Date: Wed, 30 Jul 97 10:59:57 -0400
Subject: Zeiss 25 TEM

Contents Retrieved from Microscopy Listserver Archives
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A collegue has been offered a free Zeiss 25. It would be shared between the
biology group and the materials group and the cost to them would only be
shared maintenance. He is interested in its use as a materials science TEM.

I'm looking for any information about this microscope and comments from those
familiar with it. There was no information on any TEMs at Zeiss' web sites.
Are they still producing them? What about the Omega filter?

Michael Cinibulk
UES Inc. at
Wright Laboratory
Wright-Patterson Air Force Base, Ohio
cinibumk-at-ml.wpafb.af.mil




From: Peggy Bisher :      peggy-at-research.nj.nec.com
Date: Wed, 30 Jul 1997 11:24:41 -0400
Subject: 2.3mm Grids

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Dear Newsgroup:

I need to purchase 300 mesh copper and 1000micron-slotted or hole copper
grids in the 2.3 mm size.
Does any vendor in the US supply these? I know that Agar does in the UK but
I thought it might be faster to purchase them on this side of the pond. Can
anybody out there help me? Thanks so much.

Cheers, Peggy Bisher.


NEC Research Institute
4 Independence Way
Princeton, NJ 08540.
Tel.: (609) 951-2629
Fax: (609) 951-2496
e-mail: peggy-at-research.nj.nec.com







From: Robert H. Olley :      R.H.Olley-at-reading.ac.uk
Date: Wed, 30 Jul 1997 16:33:37 +0100 (BST)
Subject: TEM and OM film

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Hello everybody,

The result of all my enquiries so far is that:

(1) there is no current manufacturer of 35mm unperforated orthochromatic
film;

(2) we have been offered a supply of some old stock at reasonable price,
but this will not last for all that long;

(3) it would be nice if we could pressurize some manufacturer into
re-doing the stuff. The Agfa Scientia appears to be the best;

(4) a gentleman from Kodak in New York sent me a roll of TX100 to try in
our optical microscope / SEM. This material allows a large range of
greyscales, and it works! It did particularly well for micrographs
between crossed polars, where objects type A are only just off extinction
and in the same field, objects type B were displaying full birefringence.
And the quality was good - "brilliant grey", if such a colour exists.

+------------------------------------------------------------------------+
| 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: Hola Chaval :      yves-at-giga.sct.ub.es
Date: Wed, 30 Jul 1997 18:57:25 +0200 (MET DST)
Subject: Re: Zeiss 25 TEM

Contents Retrieved from Microscopy Listserver Archives
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On Wed, 30 Jul 1997, Michael K. Cinibulk wrote:

} A collegue has been offered a free Zeiss 25. It would be shared between the
} biology group and the materials group and the cost to them would only be
} shared maintenance. He is interested in its use as a materials science TEM.
}
} I'm looking for any information about this microscope and comments from those
} familiar with it. There was no information on any TEMs at Zeiss' web sites.
} Are they still producing them? What about the Omega filter?

For Zeiss, see now:
http://www.leo-em.co.uk/ or http://www.mwrn.com/leo/leocont.htm

Yves MANIETTE
Universitat de Barcelona
Serveis Cientifico Tecnics
Unitat ESCA TEM
Carrer Lluis Sole i Sabaris, 1-3
E-08028 BARCELONA ESPANYA

Tel +34 3 402 16 95
Fax +34 3 402 13 98





From: Woody.N.White-at-mcdermott.com
Date: 7/29/97 3:50 PM
Subject: ISO 9001

Contents Retrieved from Microscopy Listserver Archives
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Hello James,

Our Lab is ISO certified and I run the (materials) SEM Lab.
Since ISO seems to be oriented toward a manufacturing environment
where repetitive steps are involved, I found it a "pain in the
butt" for a research/failure analysis oriented SEM facility.
Procedures is the "magic word" (translate that to lots of paper work).
I have procedures for operating the SEM, EDS, and WDS and also for
periodic calibration checks. These procedures should reflect what is
necessary to operate the equipment and generate "good" data, but at the
same time, be as general as possible. This is so that under unusual
circumstances your analysis strategies and conditions are not too
limited. Good records keeping and adherence to the procedures are
closely scrutinized. Use of ASTM methods and NIST certified standards
will "lubricate" the ISO approval procedure.

Good Luck!
Woody White
Mcdermott Technologies, Inc

______________________________ Reply Separator
_________________________________


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

I am looking for information about the ISO 9001 standard with regard to
the operation of FT-IR microscopy and SEM systems for materials analysis,
and would very much appreciate communicating with list members who have
practical experience in this area.

Many thanks in advance.

James Martin
Williamstown Art Conservation Center




From: EUGENE GORDON :      MEDJET-at-worldnet.att.net
Date: Wed, 30 Jul 1997 12:36:50 -0400
Subject: Staining for Spurr's

Contents Retrieved from Microscopy Listserver Archives
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Hello all,

Is anyone familiar with stains and protocols for tissues embedded in
Spurr's low viscosity resin? I have been using toluidine blue on one
microns and need to achieve a greater differentiation among collagen fibers
and keratocytes within the stroma of the human cornea.

Thanks for any suggestions,

Dan Caruso c/o Eugene Gordon





From: Dan Kaszubski :      danzk-at-cysource.com
Date: Wed, 30 Jul 1997 14:13:52 -0500
Subject: Re: Welcome, Rules, FAQ Docs - Microscopy ListServer

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} -------------------------------------------------------
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} ------------------------------------------------------
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} Not any longer. For a time this Listserver automatically forwarded all
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} ---------------------------------------------------------------
} ------------
} I think I need to test the Email, what do I do?
} -----------------------------------------------
} ----------------------------
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} instructions, then
} your Email program must be working. So no further testing is really
} necessary.
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} copies
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}
} CompuServe, AOL, GENIE, etc....) We have a lot of them, and this
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} If you really feel you must test your Email Package or think you need
} help
} then send a message to:
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} He's sometimes a patient soul, who at times has been known to read his
} Email
} and occasionally answers questions. Especially, late at night.
}
} --------------------------------------------------------------
} -------------
} Is there a Digest Mode?
} -----------------------
} ----------------------------------------------------
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} Digest Mode is not available on the present server. It is an option
} which
} will be added in the future.
}
} Digest Mode compresses all message from a single day into one long
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} which is then sent to all subscribers. It is a nice option, and I
} intend to
} get it running eventually.
}
} --------------------------
} -------------------------------------------------
} When I send a message I get (sometime many) Bad/Bounced Messages back
} why?
} ----
} -----------------------------------------------------------------------
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} ------------------------------------
} ---------------------------------------
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} subscribers from more than 40 countries to this list.
}
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} --------------------
} -------------------------------------------------------
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} http://www.msa.microscopy.com/MicroscopyListserver/Micro
} copyArchives.html
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}
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} -----------------------------
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} can
} always buy him a beer or two. He's usually easy to recognize, and his
} arm
} twists easily.
}
} --------------
} -------------------------------------------------------------
} Who do I contact with problems?
} -------------------------------
} --------------------------------------------
}
} God-at-Garden.Eden.Earth.Com
}
} -------------------------
} --------------------------------------------------
} Who Runs this?
} --------------
} -------------------------------------------------------------
}
} This listserver is run by Nestor J. Zaluzec
} (Zaluzec-at-MSA.Microscopy.Com) It
} has been & continues to be run/administered/babied mostly in his spare
} time
} and usually late in the evening or earlyhours of the morning.
}
} -------------------------------------------------------------
} --------------
} Special Announcement/News
} -------------------------
} --------------------------------------------------
}
} At the 1995 Winter Council Meeting, the Microscopy Society of America
} (MSA)
} approved a proposal to support this server as well as other
} telecommunications options as a FREE Service to ALL
} persons/organizations
} worldwide who are involved or interested in Microscopy and/or
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}
} Thanks to this support, the listserver will be upgrading the hardware
} software and services for use by Microscopist's/Microanalysts
} Worldwide.
}
} ----------
} -----------------------------------------------------------------
} Is there an FTP and/or WWW site?
} --------------------------------
} -------------------------------------------
}
} Yes, both an FTP and WWW sites exist here are the addresses:
}
} WWW http://www.msa.microscopy.com
}
} FTP ftp.msa.microscopy.com (Anonymous Login Enabled)
}
} *********************************
} Last Updated April 15, 1997
} Nestor J. Zaluzec
} Your Friendly Neighborhood SysOp.
} *********************************
} End of File
} *********************************








From: Lee_ccmail_Wagstaff_at_IRV006-at-ccmailgw.mcgawpark.baxter.com
Date: Wed, 30 Jul 1997 13:06:43 -0500
Subject: Looking for used sputter coater

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

A friend who runs a small lab in town needs a fully operational
sputter coater (for cheap of course). Anyone out there have any leads
I can pass on?

Contact me directly at wagstale-at-baxter.com

Thanks All.




From: Maurice Smith :      micscape-at-netlink.co.uk
Date: Wed, 30 Jul 1997 23:42:43 +0000
Subject: help us!

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

I'll make it brief: please drop by and see what we have bean up to for
the last 2 years to promote study through Amateur microscopy.

You just might find it refreshing to know we are out here and going for
bust!


http://wwww.microscopy-uk.org.uk


People who care. Non-commercial. Non-profit-making.

20,000 hits a week means others have found us!

Thanks for your time. I won't bother you again thru this means.
Check us out once just once only to see if this intrusion was warranted.


my regards,

Maurice




From: David Griffiths :      DGriffiths-at-SAS.Samsung.com (by way of Nestor J.
Date: Wed, 30 Jul 1997 19:47:22 -0500
Subject: Position Available -- Entry Level TEM Technician

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

}
} The Company:
}
} Samsung Austin in Austin, Texas wants to offer you more than a job. We want
} to offer you the chance to develop a career. Becoming involved in a
} fast-paced start-up operation is both exciting and challenging and we look
} for employees who are energetic, flexible, and team-oriented.
} Are you willing to go the extra mile? Are you comfortable with change? Are
} you interested in great benefits? Are you excited about working in a start-up
} environment? Are you interested in a career where you will be an integral
} part of a new company? Are you comfortable making decisions that will
} influence the development of our corporate culture?
} If the answer to these questions is "yes," then Samsung Austin is the company
} for you.
} For more information visit our web site at www.sas.samsung.com
}
}
} The Position
}
} We are currently looking to hire an Entry Level TEM Technician. The
} requirements for this position is simply an Associate Degree in a technical
} major. Experience is preferred but not necessary. The position will involve
} shift work.
}
} If someone know of a person who would consider this position or if you know
} of a 2 year college that offers courses on TEM Sample Prep please feel free
to contact me. We are going to start interviewing next week so please
} get your resume to me quickly.
}
} Resumes can be faxed or emailed to:
} David A. Griffiths
} Fax (512)491-1165
} Pager (512)209-4132
} e-mail DGriffiths-at-sas.samsung.com
}






From: lag-at-pegasus.cc.ucf.edu (Lucille A. Giannuzzi)
Date: Wed, 30 Jul 1997 21:53:23 -0400
Subject: SEM backscattered patterns

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

Thanks to all for your help! We were able to get a pattern after tweaking
the specimen quality.

**************************************************************************
Lucille A. Giannuzzi, Ph.D. phone: 407 823-5770
University of Central Florida fax: 407 823-0208
Dept. of Mechanical, Materials, and Aerospace Engineering
PO Box 162450
4000 Central Florida Blvd.
Orlando, FL 32816-2450 email: lag-at-pegasus.cc.ucf.edu
**************************************************************************






From: Garber, Charles A. :      cgarber-at-2spi.com
Date: Wed, 30 Jul 97 23:26:34 -0500
Subject: 2.3 mm grids availability

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

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

Peggy Bisher wrote:
=================================================
I need to purchase 300 mesh copper and 1000micron-slotted or hole copper
grids in the 2.3 mm size. Does any vendor in the US supply these? I know
that Agar does in the UK but I thought it might be faster to purchase them
on this side of the pond. Can anybody out there help me? ==================
================================
Because of such little demand for the 2.3 mm size, in North America, they
not found as readily here as they are in Europe. However, you can find them
in a few mesh sizes on our website under "regular" grids, "micron" style.
Other mesh sizes and types are available also but are not yet up on the
website. I think that some of the other EM suppliers of consumables in the
USA offer 2.3 mm grids as well, or at least they did until recently.

Chuck

===================================================
Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400
President 1-(800)-2424-SPI
SPI SUPPLIES FAX: 1-(610)-436-5755
PO BOX 656 e-mail: cgarber-at-2spi.com
West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com


Look for us!
############################
WWW: http://www.2spi.com
############################
==================================================







From: Mark E. Darus (216) 266-2895 General Electric Co. :      darus-at-cle.dnet.ge.com
Date: Wed, 30 Jul 97 14:17:43 EDT
Subject: Beam Temperature

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

I'd like to examine a sample in my SEM. My concern is that if the beam
heats the sample, it may volatile and contaminate my column. I don't have
a cold finger. Does the beam heat the sample? What is the temperature of
the beam? What sort of elevation in temperature does a sample go through
during an examination? What about different accelerating voltages, do they
produce beams with different temperatures?
Thanks
Mark E. Darus




From: Ten Brink G. IEG CP :      BRINK-at-skn.sc.philips.com
Date: Thu, 31 Jul 1997 08:53:19 CET-0100
Subject: request

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

Request

Dear colleagues and calibration standard manufacturers.

A committee within the Dutch Microscopy society is working on the
general issue of calibration. In June we made a request on the
microscopy listserver. The only helpful item that came up was a
reference for the NIST calibration standard SRM484 G (which I already
own).

On the Internet there is however a large variety in calibration
standard available. It's a pity that so little people have responded
therefore one can only assume that they are:
1. not using the standards
2. not interested in obtaining accurate results to satisfy the ever
demanding customer
3. not on the Internet
4. don't feel the need to talk to us (were told not to by there boss
or were shy)
5. under the impression we're not nice persons 8(

Therefore I see no alternative then to ask you again a few questions.
The questions are intended for users of a SEM and the manufacturers
of calibration standards but anyone who has some interesting things
to say is welcome.

Are you all aware of the writing of Herr Clive Walker on "The Report
on the 4th Plenary Meeting of ISO/TC202 Londen 6-9 th May 1997" ??.
In this writing there are still some issues that must be dealt with.
One issue is calibration but unfortunately a consensus was not
reached and the issue remains open. So, let's say I want to apply
for a STER-lab certificate; what issues are to be dealt with other
than calibration (other than the references given in the NIST
calibration routine).

Some questions that come to my mind are:
* Standard defining terms used in Micro Beam analysis.
* General machine settings ?
* Info about the mounting size.
* Is the sample 1D or 2D (the NIST standard is a 1Dimension standard,
IBSEN has a 2D standard) are they the same in the sense that they can
both be used for the same calibration routine (for instance the ASTM
E 766-93).
* Is it important to know about the difference in Z (atom number).
The NIST standard has thin gold lines in a nickel base. Others use a
silicon grid. Does the difference in Z also mean that there might be
a difference in signal to noise ratio when the same machine offsets
are used.
* Can you give us some photograph's of the calibration sample at
let's say 3 different magnifications, 3 different accelerating
voltages and corresponding spotsizes.
* Can you give us detailed information about the accuracy of the
calibration standard and a normal achievable level of accuracy with
an average SEM (knowing it's hard to define a normal SEM) including
the methods used for statistical proceedings eg. .
* Do you have references from laboratory that have a STERlab
certificate(not ISO 9000) or equal and I refer to the new ISO/IEC
guide 25 using your calibration standards.

I know of the following calibration standard manufactures:
Ernest F. Fullam, Inc.
SPI.
Energy Beam sciences Inc.
NIST.
MOXTEK, order at Ted Pella, Inc.
TCL (I am not sure if they make standards but they do SEM calibration)
IBSEN
MAG*I*CAL (National Research Council of Canada)

All info is derived from the Internet using common search engines.
The list is probably not complete and I would appreciate it if the
list can be made up to date with more info.

It is not my intention to give a rating for the calibration standards
that are provided with a large variety in spec's and cost. What I
(we) want is to give people who are on the road for ISO/IEC
certification some info about the calibration standards and the
issues that come along in the process.
Therefore I give everybody a equal opportunity to give info about
his or her calibration sample making it more easy for us people to
make a choice in what calibration standard the most suited is for his
or her specific use.

As stated above this request goes to all the known manufacturers on
the list and this request will also be posted on the Microscopy list
server.

Many thanks in advance,

Gert ten Brink
Philips Semiconductor BV.
Fysisch analyst
postbus 10
9500AA Stadskanaal
tel 0599632380
fax 0599632505
e-mail brink-at-skn.sc.philips.com
privat asslab-at-xs4all.nl

Ps. all cost made and found reasonable (photo material, stamps etc.)
will be refunded.
On the other hand: if this leads to a large rise in sales in
calibration standards can I give you the number of my bankaccount ?

Adresses where the questions will be asked:

Microscopy Listserver
Microscopy-at-MSA.Microscoppy.Com

MAG*I*CAL
South Bay Technology, Inc.
t.a.v David Hendriks
1120 Via Callejon
San Clemente, CA

Micro-World
mwrn-at-mwrn.com

Microscopy Standards from Ernest F. Fullam, Inc.
900 Albany Shaker Rd. Latham, New York 12110
Sales-at-fullam.com

Probing & Structure
p&s-at-ultra.net.au


Energy Beam Sciences, Inc.
P.O. Box 468
11 Bowles Road
Agawam, Massachusetts 01001
ebs-at-ebsciences.com

MOXTEK, Inc.
452 West 1260 North Orem UT 84057
order at Ted Pella, Inc.
gstewart-at-moxtek.com

TCL
nog opzoeken

IBSEN Micro Structures A/S
Calibration standards
ibsen-at-risoe.dk

SPI Structure Probe, Inc.
e-mail opzoeken

NIST
t.a.v. Thomas E. Gills
Gaithersburg, Maryland, MD 20899

Allen R. Sampson
Advanced Research Systems
St. Charles, Illinois
ISO/IEC Guide 25-draft four
Recommendations for implementation of ISO Quality Standards in an SEM
laboratory
Both can be found on the Internet

Other interesting items,
SCANNING
Vol. 18, 533-538 (1996)
Vol. 18, 539-555 (1996)
Vol. 17, 287-295 (1995)
Vol. 18, 1-7 (1996)
Vol. 18, 50-54 (1996)

Met vriendelijke groet,

Gert ten Brink
Fysisch Analist
Philips Semiconductor B.V.
Stadskanaal IEG-lab geb. Ao-p
tel. 0599632380
fax. 0599632505
e-mail: brink-at-skn.sc.philips.com
privat: asslab-at-xs4all.nl






From: Krzysztof Jan Huebner :      hubner-at-czapla.IOd.krakow.pl
Date: Thu, 31 Jul 1997 09:11:16 +0200 (MET DST)
Subject: e-mail to swedish institue for metals research

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



Good morning to all,

I need e-mail to Swedish Institute for Metals Research - Stockholm


best regards for all



Krzysztof Jan Huebner

{hubner-at-IOd.krakow.pl} :-)

FOUNDRY RESEARCH INSTITUTE
Research Materials Department
Structural and Physical Research Laboratory

Zakopianska 73 Call +48 12 605022 ext. 356
30-418 KRAKOW - POLAND Fax +48 12 665478, + 48 12 660870







From: jrosato-at-ascella.net
Date: Thu, 31 Jul 97 00:19:46 EST
Subject: Software boosts profits by 50% !!

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

ATTN: Business Owner/Internet Marketer:

Your profits are about to increase by 50%, practically overnight! If you are selling any type of product or service, on-line or off-line, this software is an absolute MUST for your business. Please read on...

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(You can also enter our online sweepstakes to WIN a Pentium laptop PC!)
(Note: Our server is in the process of an upgrade, so if you experience any difficulties accessing our web site, please try again later.)

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- Simple. INCREASED REVENUES - by up to 50% or more! More and more businesses are adopting this new system of accepting payment from their customers. They have all experienced INCREASED PROFITS by providing a new medium for their non-credit card paying c
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- As you are probably aware, credit cards invoke IMPULSE SHOPPING - you get the customer when they're "HOT". As you are also probably aware, obtaining a credit card merchant account for a small or home-based business can be very difficult and very expensi
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- Then go to your bank and deposit the check(s), just as you would deposit any other checks. That's it - instant money!

* Note to non-U.S. residents: Checks Xpress cannot be used to debit non-U.S. bank accounts; however, if you're doing business internationally, you can still use Checks Xpress to accept payment from all U.S. based consumers and businesses, provided that yo
ur financial institution accepts U.S. checks.

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- Absolutely! The following federal regulations make this all possible and 100% legal: "Uniform Commercial Code, Title 1, Section1-201(39) and Title 3, Sections 3-104, 3-401, and 3-403; Code of Federal Regulations, Title 12, Chapter II, Part 210 and Reg
ulation J, Federal Reserve Bank, Part 2, Sections 4a-201 to 4a-212. Only verbal agreement is required for authorization. See Romani versus Harris 255 Md. 389."

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- No special bank account or authorization is required - your current bank account is already able to accept bank drafts! In addition, no signature is required to deposit and process the check (refer to the federal regulations listed above). As long as
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- Also, no special magnetic ink toner is required for your printer. Since most banks are using the optical (laser beam) check reading system, ink toner is not an issue. For those banks that might still be using the older, magnetic check reading system, yo
ur check will still be processed manually. Either way, your Checks Xpress checks are as good as gold!

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- Simple, friendly user interface. No special computer skills required. Checks Xpress was created with the most novice of computer users in mind. In fact, Checks Xpress has been rated as the EASIEST to use check printing program, ever. Installation is a b
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- Store an unlimited number of your customers' checks on your computer, for archival, future retrieval, and monthly account debiting.

- Automatic check number incrementing feature allows you to automatically maintain sequential check numbers for better record keeping and less data entry.

- Custom signature line feature allows you to customize the information that appears on the check's signature line, or use the built-in authorization information along with a date and time stamping feature.

- Custom printer alignment. Easy alignment feature ensures that Checks Xpress will print perfect checks every time with your laser or inkjet printer.

- Free tech support should you ever run into any difficulties with Checks Xpress.

- Checks Xpress is not to be confused with the accounting or financial software programs available in stores (Quicken, Peachtree Accounting, etc.). Unlike those programs, Checks Xpress prints the special bank codes found along the bottom of every check. T
he store-bought programs do not print the special bank codes, and they are designed to only print checks from the same checking account.

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- Any computer system running Windows95 or Windows NT 3.51/4.0
- A laser or inkjet printer
- 3.5" floppy disk drive
- 5 MB free hard drive space
- A standard checking account

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*** SPECIAL OFFER *** If you order within 10 days of receiving this offer, you will be eligible for a $40 INSTANT ONLINE REBATE! Your total cost to own Checks Xpress WILL BE ONLY $99.00! We GUARANTEE you will not find a better check printing system for le
ss! To qualify for your $40 discount, you will need to provide a valid Instant Online Rebate (IOR) number when you place your order.
* IMPORTANT * DO NOT LOSE THIS NUMBER - your valid IOR number is: CXP6410

-----------------------------------------------------------------------------
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- Absolutely! You can try Checks Xpress AT NO RISK! After you have tested Checks Xpress in demo mode to your complete satisfaction, simply contact out technical support department to obtain your valid "unlock code", which will permanentely remove all limi
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US REFUND, NO QUESTIONS ASKED.

-----------------------------------------------------------------------------
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-------------------------------- cut here -----------------------------------
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ZR ONE Technologies
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#2. FAX: Print this order form and either (a.) tape your voided personal/business check below this line, or (b.) fill in your credit card information in the spaces above, and fax your order to: 408-260-7811

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For information about our Authorized Reseller (AR) program, visit: http://205.199.2.39/ZR1/reseller.htm

Thank You for your order! We appreciate your business and we look forward to serving you again in the future.

Copyright (c) 1997 ZR ONE Technologies. All rights reserved.





From: Ian MacLaren :      I.MacLaren-at-BHAM.AC.UK
Date: Thu, 31 Jul 1997 10:37:32 +0100
Subject: Re: Beam Temperature

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

Dear all,
I am interested in this question, I sometimes see evidence of local heating
of samples during TEM observation but I have never had much idea about
quite how much heat is generated by the electron beam. Does anyone know or
know how to find out?

Also, does anyone have any idea why oily blobs (obviously from oil in the
vacuum system) sometimes appear on the sample in the very place that you
are observing (as opposed to any other place on the sample)?

Thanks


++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Ian MacLaren, Tel: (44) (0) 121 414 3447
IRC in Materials for FAX: (44) (0) 121 414 3441
High Performance Applications, email: I.MacLaren-at-bham.ac.uk
The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/
Birmingham B15 2TT,
England.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++






From: James Martin :      James.S.Martin-at-williams.edu
Date: Thu, 31 Jul 1997 07:28:06 -0400 (EDT)
Subject: Re: ISO 9001

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

Many thanks to all who responded to my inquiry re ISO 9001. Your
assistance has been helpful.

James Martin







From: Larry Stoter :      LPS-at-teknesis.demon.co.uk
Date: Thu, 31 Jul 1997 08:44:44 +0100
Subject: Re: Beam Temperature

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

} I'd like to examine a sample in my SEM. My concern is that if the beam
} heats the sample, it may volatile and contaminate my column. I don't have
} a cold finger. Does the beam heat the sample? What is the temperature of
} the beam? What sort of elevation in temperature does a sample go through
} during an examination? What about different accelerating voltages, do they
} produce beams with different temperatures?
} Thanks
} Mark E. Darus

The beam certainly can heat some specimens to a sufficiently high
temperature to cause evaporation. The temperature reached is going to
depend on all sorts of things - specimen thermal conductivity, how well
specimen is connected to mount, how well mount is connected