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From: zaluzec-at-aaem.amc.anl.gov
Date: Fri, 1 Jan 2010 09:23:03 -0600
Subject: [Microscopy] Administrivia: Happy New Year - 2010

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Happy New Year Colleagues;

Welcome to the 18th year of operation of the Microscopy ListServer
a free service to the world wide microscopy community, sponsored
jointly by your Friendly Neighborhood SysOp and the Microscopy Society
of America.

It was another productive year for all of us. During 2009, the
ListServer
delivered 2270 messages to ~ 3000 subscribers (} 325 Gb of Email)
around the world, with only minimal hassels for most of you (that I
know about).

The complete Microscopy ListServer Archives for 2009-1993 are on-line
at

http://www.microscopy.com.

I have also just resurrected the Surplus Equipment Listings. That
service ( which can be used to advertise equipment for sale vs
equipment to be given away) was previously located on the MSA WWW
site and was discontinued. You can now find the Surplus Equipment
listings
under the On-Line DataBases option along with
WWW site and Meeting/Conference registrations at the URL:

http://www.microscopy.com

As always if you have questions about suitability of postings or
are having problems, feel free to contact me at (zaluzec-at-microscopy.com)

Cheers,

Nestor
Your Friendly Neighborhood SysOp


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From: lcgould-at-med.cornell.edu
Date: Mon, 4 Jan 2010 15:35:04 -0600
Subject: [Microscopy] EMI cancellation systems

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Hi Listers and Happy New Year.

I have been lucky. We have obtained an FEI Quanta 600 SEM with an
environmental chamber from an affiliated institution. This being New
York City where space is always at a premium, we located a room, near
enough to the rest of my facility but which has some EMI field issues.
I've gotten information from a number of manufacturers and I am
trying to make a well-informed decision about what I get: type
(cage vs whole room) as well as vendor.
May I ask those of you with such systems to respond to me off-list
about what you have?
Please answer the following questions:
Which system?
What vendor?
How long have you had it?
Has it been reliable?
Is it easy to use?
Are you happy with your system?

thanks much!
Lee

--
Lee Cohen-Gould, M.S.
Sr. Staff Associate in Biochemistry and
Cell & Developmental Biology
Director, Electron Microscopy & Histology
and Optical Microscopy Core Facilities
Weill Cornell Medical College

voice (212)746-6146
fax (212)746-8175
http://www.med.cornell.edu/research/rea_sup/
http://www.cornellcelldevbiology.org
http://www.cornellbiochem.org

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From: news-at-emsdiasum.com
Date: Mon, 4 Jan 2010 17:14:58 -0600
Subject: [Microscopy] viaWWW: Aurion ImmunoGold Silver Staining Workshop

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Email: news-at-emsdiasum.com
Name: Sue Brandom

Organization: Electron Microscopy Sciences

Title-Subject: [Filtered] Aurion ImmunoGold Silver Staining Workshop

Question: Aurion ImmunoGold Silver Staining Workshop
University of Florida College of Medicine
Feb 1-3, 2010

Aurion and Electron Microscopy Sciences will hold their workshop on
Immuno Gold Silver Staining at the University of Florida College of
Medicine from February 1 to 3, 2010. The course objectives are to
provide researchers with the opportunity to learn the theory and
practice of immunogold labeling and to promote technology exchange
and research collaboration. Participants are encourage to bring their
own specimens to process under expert guidance.

For more information contact:
Stacie Kirsch
stacie-at-ems-secure.com
215-412-8402
www.emsdiasum.com/microscopy


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From: oshel1pe-at-cmich.edu
Date: Tue, 5 Jan 2010 11:51:04 -0600
Subject: [Microscopy] Re: SEM of cancer cells

Contents Retrieved from Microscopy Listserver Archives
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Erman,

I've done cancer cells for SEM, although not on carbon nanotubes.
Fixation & processing was:
1 hour in 1.25% glutaraldehyde in 0.1 M PO4 buffer + 1% monomeric
tannic acid, room temperature
[optional: wash in buffer, then post-fix in 1% OsO4 in either water
or buffer -- usually didn't need this step]
dehydrate through ethanol; started at 30%, but may need to start at
10 or 15% depending on your cells, then in 10% steps through 90%,
then 95%, then 3 X 100%. 5 minute steps, assuming non-confluent
monolayer of cells.
Then critical point dry, 3 soaks of 5 minutes each, 5 soaks may be needed.
Note: on CNTs ... critical point drying may affect how the cells sit
on the tube forest. I have no good idea, just a feeling CPD will be
OK.
But.
HMDS (hexamethyldisilizane) drying may work better here. After the
final 100% ethanol, go 1:1 absolute ethanol:HMDS then 3 X 100% HMDS,
10 minutes each, blot of excell fluid *but leave samples just
covered* and air dry.
NOTE: HMDS *must* be used in a fume hood.

Mount & sputter coat as usual.

Phil

} Dear All,
}
} I would like to find out if there is an easy and effective procedure which
} would allow me to prepare samples of cancer cells grown on CNT forests for
} observation under SEM. I will be using an CZ EVO 40 (LaB6 filament) and I
} have never worked on soft materials.
}
} Happy holidays and new years to you all
}
} Erman Bengu
}
} =================================
} Erman Bengu
}
} Assistant Professor of Chemistry
} Department of Chemistry
} Bilkent University
}
} Mailing Address:
} Bilkent University,
} Department of Chemistry,
} 06800, Bilkent, Ankara
} Turkey
}
} Office: SB #311
} E-mail: bengu_AT_fen.bilkent.edu.tr
} Phone (Office): +90 (312) 290-2153
} (Lab1): +90 (312) 290-2663
} (Lab2): +90 (312) 290-3332
} Fax: +90 (312) 266-4068
} Web: http://www.fen.bilkent.edu.tr/~bengu
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: bozzola-at-siu.edu
Date: Wed, 6 Jan 2010 15:01:13 -0600
Subject: [Microscopy] EM: LN2 cooled versus electronically cooled EDS detectors

Contents Retrieved from Microscopy Listserver Archives
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I am considering the advantages/disadvantages of the traditional,
LN2-cooled EDS detectors versus the Peltier or electronically cooled
detectors.

One huge advantage is obvious: no LN2 filling needed. But, I am
concerned about the relative longevity (durability) of the
electronically cooled system versus the tried-and-true LN2 systems.
Anyone care to comment on this?

Also, with the Peltier cooled units, can one cycle the cooling on when
needed, and then back off when finished?

Thanks.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

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5, 15 -- Subject: EM: LN2 cooled versus electronically cooled EDS detectors
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From: gary-at-gaugler.com
Date: Wed, 6 Jan 2010 15:59:08 -0600
Subject: [Microscopy] Re: EM: LN2 cooled versus electronically cooled

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The Peltier cooled SDD EDS units are totally awesome.

There are only a few specific applications that Si(Li)
is better than SDD. The current generation of SDDs
are terrific. Yes, you can turn them off when not
needed and on when needed. Most makers' models
cool down for use in minutes if you want to
work that way. You can also shut off the EDS PC
if desired. The total power drain for the Peltier
cooler controller and PC is quite low. I figure
that a typical Peltier controller uses about 20 Watts
when temperature is stable, but this depends on whether
the Peltier is a single, dual or triple stack. The small
area SDDs are usually single Peltier. My current 40mm^2
SDD typically runs at about 2.5V and 4A for the Peltier
stack.

About the PC: These are dual or quad core of at least
2.5GHz with 2GB-3GB DDR3 DRAM. In order to process
the higher counts, a faster PC is needed, otherwise,
the performance benefits of the SDD are lost.

The SDD units are very reliable, small and
able to produce high cps. The newer DPPs also
are able to process these higher cps values
if it is in a high performance PC. A real
decision is whether to get the new PC from the
EDS maker pre-loaded with WinXP Pro or Win7. As
you know, moving to Win7 from XP cannot be done
unless Vista is installed over XP first. Personally,
I would not get Win7 until users have debugged it
to a lower hassle factor. Some EDS softwhere (I like
that catchy term) will handle 64-bit OS while others
may stick with 32-bits for now. I don't know if there
is any significant difference in throughput.

The down side is that your legacy EDS system will
have to all be gone...nothing remains of the old
system (detector, DPP, PC or software). For EDAX
software, the Genesis program is still used but
requires a newer version which is included in the
new SDD package of items. This is probably the
same situation for other makers.

The last item of interest is the digital scan generator.
Some makers may be able to use the old one. EDAX
has an SG-II which captures one channel of video up to
6400x4000 pixels and a SG-III which is the same resolution
but can capture two channels at the same time. This is
handy for capturing SE and BSE at the same time and in
perfect sync and alignment.

gary g.


At 01:03 PM 1/6/2010, you wrote:

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From: krivanek-at-nion.com
Date: Thu, 7 Jan 2010 18:46:39 -0600
Subject: [Microscopy] viaWWW: position available at NION

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Email: krivanek-at-nion.com
Name: Ondrej Krivanek

Organization: Nion Co.

Title-Subject: [Filtered] position available

Message: Nion R&D (www.nion.com/about.html) has
an immediate opening for an instrumentation
specialist to work on developing a high-energy
resolution monochromated scanning transmission
electron microscope (STEM) system. The
instrument will use a monochromator/chromatic
aberration corrector of a radically new design
(Krivanek et al., Phil. Trans. R. Soc. A vol 367
(2009) 3683-3697), in a collaborative project
with Arizona State university. It promises to
revolutionize electron microscopy and electron
energy loss spectroscopy (EELS) by combining meV
level energy resolution with Angstrom-level spatial
resolution, as well as improved spatial
resolution at low primary energies.

The specialist will work within the Nion design
team on all aspects of the monochromator project,
from evaluating the electron-optical properties
of candidate systems, to mechanical and
electrical design, and the construction, testing,
and demonstration of the systemís performance on
key materials problems. The ideal candidate will
have an outstanding research record and be deeply
interested in electron-optical instrumentation
and its applications.

Nion offers an informal, friendly and efficient
working environment with a track record of
producing revolutionary instruments that is
second to none, and a competitive salary and
comprehensive benefits. Send your resume and an
electronic copy of your two principal
publications to Dr. O.L. Krivanek
(krivanek-at-nion.com).


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From: mjamison-at-caissonlabs.com
Date: Thu, 7 Jan 2010 18:47:11 -0600
Subject: [Microscopy] viaWWW: thin section then GUS staining

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Email: mjamison-at-caissonlabs.com
Name: Michelle Jamison

Organization: plant labratory

Title-Subject: [Filtered] thin section then GUS staining

Question: Hi all
Has anyone done GUS (B-glucuronidase) staning following embedding and
sectioning with paraffin? I have done whole tissue staining with
good luck, but need to get into maturing seeds of grain crops to see
expression. I am thinking that Those are thick and hard to penetrate.
Thank you for your help.
Michelle

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==============================Original Headers==============================
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From: brantnerc-at-nbacc.net
Date: Fri, 8 Jan 2010 08:17:50 -0600
Subject: [Microscopy] viaWWW: EM research assistant position open at BNBI/NBACC

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Email: brantnerc-at-nbacc.net
Name: Christine Brantner

Organization: Battelle National Biodefense Institute/NBACC

Title-Subject: [Filtered] EM research assistant position open at BNBI/NBACC

Question: There is an immediate opening for a research assistant in
electron microscopy in Frederick MD at Battelle National Biodefense
Institute. The applicant should have basic specimen prep skills for
TEM and SEM. The position works with investigators to identify,
characterize and analyze a variety of viral, bacterial, fungal and
animal tissue samples. Experience with electron microscopes and all
accessory EM equipment is a plus. The successful applicant will be
working in an exciting new lab where the mission is to help protect
the nation against bioterrorism. The National Biodefense Analysis and
Countermeasures Center is located at Fort Detrick, and is a federally
funded research and development center for Dept of Homeland Security.
See requirements for employment in the posting below.

For more information, please see the following link.
http://www.bnbi.org/careers.html (Research Assistant, Electron
Microscopy #192)

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From: Michal.Jarnik-at-fccc.edu
Date: Fri, 8 Jan 2010 08:42:32 -0600
Subject: [Microscopy] Balzers parts free to good home

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

Our old Balzers freeze-etcher/evaporator is being retired. I am offering
the free-standing Balzers control units to any interested party willing
to arrange (and pay) for shipping. The instruments are located in
Philadelphia, PA, and were functional last time I used them (about 2
years ago).

Quartz Crystal Thin Film Thickness Monitor
Freeze Etching Device Control Unit GA-1
Control Unit EVM 052 (high voltage control)

If interested, please contact me.

Thanks,

--

Michal Jarnik, Ph.D.
Fox Chase Cancer Center
Philadelphia, PA



==============================Original Headers==============================
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From: TindallR-at-missouri.edu
Date: Fri, 8 Jan 2010 10:43:48 -0600
Subject: [Microscopy] EM imaging: Self-censorship?

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

The list has been a little slow lately, so here's a topic for potential discussion:

On occasion we look at things in our scopes that have no good basis for reference---no previous publications, no other EM images to compare with---you get the picture. My method has been to take representative images of what is there, even if the images have a wide variety of things in them that don't resemble each other or what the sample supposedly "should" look like.

It's that "should" that is the problem. It sometimes happens when we send these images to the clients that they grab onto whatever looks like what they want to see, pretty much ignore anything else, then starting making assertions about the images that go waaaay beyond what the image can support and want to plug all that into a publication. (I've had people get all Eureka! about the "champagning" artifact on a negatively-stained prep, for example.)


So the question is, if the EM operator has a reasonable suspicion, but not a certainty, that an image is showing artifact or something that is not really the part of the sample the researcher wants to see, how should that be handled? Should we send the images along with our caveats and risk having them having them published with interpretations that go beyond the data and may just be dead wrong? Or should we self-censor and not send these images?

Remember, I'm not talking about things that we KNOW are artifact or garbage. That's a clear call. I'm talking about imaging things that may not have been seen before, and nobody really knows what they look like (but they think they do), thereby making it difficult to separate artifact from real data. What we do now is send the images with our comments and hope that the client isn't so desperate for a publication that they ignore our cautions. We are virtually never listed as co-authors so that's not really an issue, but still.....I like clean science.

How do other members of the Collective handle these cases?

Cheers and stay warm!

Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




==============================Original Headers==============================
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From: baskin-at-bio.umass.edu
Date: Fri, 8 Jan 2010 10:47:06 -0600
Subject: [Microscopy] Re: thin section then GUS staining

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

Michelle,
The best images I have seen in the literature of Gus stained
sections are from material embedded in technovit. This is (I think)
glycol methacrylate, which is friendly to water soluble things. It is
also possible that PEG sections would work, but I have not seen
images. Organic solvents tend to remove the GUS reaction product. It
definitely won't work in butyl methyl methacrylate (we tried) and
unlikely to work with epoxies. In these scenarios, the staining is
done first and then everything is embeded and sectioned. I expect
that the GUS enzyme would loose activity after fixation and
embedding, even in paraffin. I don't know that for sure. And perhaps
it could be tissue specific. For a tried and true method, I would
look at Technovit. Ben Scheres lab have used this successfully as
have many others.
Good luck,
Tobias


} ---
}
} Email: mjamison-at-caissonlabs.com
} Name: Michelle Jamison
}
} Organization: plant labratory
}
} Title-Subject: [Filtered] thin section then GUS staining
}
} Question: Hi all
} Has anyone done GUS (B-glucuronidase) staning following embedding and
} sectioning with paraffin? I have done whole tissue staining with
} good luck, but need to get into maturing seeds of grain crops to see
} expression. I am thinking that Those are thick and hard to penetrate.
} Thank you for your help.
} Michelle
}
} Login Host: 129.123.46.133

--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Biology Department
/_ / __ /__ \ \ \__ 611 N. Pleasant St.
/ / / \ \ \ University of Massachusetts
/ / / \ \ \ Amherst, MA, 01003
/ / ___ / \ \__/ \ ____
www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

==============================Original Headers==============================
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4, 21 -- From: Tobias Baskin {baskin-at-bio.umass.edu}
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From: baskin-at-bio.umass.edu
Date: Fri, 8 Jan 2010 10:52:25 -0600
Subject: [Microscopy] Re: thin section then GUS staining

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

Michelle,
The best images I have seen in the literature of Gus stained
sections are from material embedded in technovit. This is (I think)
glycol methacrylate, which is friendly to water soluble things. It is
also possible that PEG sections would work, but I have not seen
images. Organic solvents tend to remove the GUS reaction product. It
definitely won't work in butyl methyl methacrylate (we tried) and
unlikely to work with epoxies. In these scenarios, the staining is
done first and then everything is embeded and sectioned. I expect
that the GUS enzyme would loose activity after fixation and
embedding, even in paraffin. I don't know that for sure. And perhaps
it could be tissue specific. For a tried and true method, I would
look at Technovit. Ben Scheres lab have used this successfully as
have many others.
Good luck,
Tobias


} ---
}
} Email: mjamison-at-caissonlabs.com
} Name: Michelle Jamison
}
} Organization: plant labratory
}
} Title-Subject: [Filtered] thin section then GUS staining
}
} Question: Hi all
} Has anyone done GUS (B-glucuronidase) staning following embedding and
} sectioning with paraffin? I have done whole tissue staining with
} good luck, but need to get into maturing seeds of grain crops to see
} expression. I am thinking that Those are thick and hard to penetrate.
} Thank you for your help.
} Michelle
}
} Login Host: 129.123.46.133

--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Biology Department
/_ / __ /__ \ \ \__ 611 N. Pleasant St.
/ / / \ \ \ University of Massachusetts
/ / / \ \ \ Amherst, MA, 01003
/ / ___ / \ \__/ \ ____
www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

==============================Original Headers==============================
4, 18 -- From baskin-at-bio.umass.edu Fri Jan 8 10:52:25 2010
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4, 18 -- Subject: Re: thin section then GUS staining
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From: Elliott-at-arizona.edu
Date: Fri, 8 Jan 2010 11:08:05 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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

Good question Randy

For my part, I have two hats. I am a faculty researcher and a person
who directs a service facility.

As a faculty member I am a bit pushy about things I know about. I
know exactly what you are saying about people latching on to something
they like and ignoring the rest. I push rather hard for a more
neutral reading of the data. When we have done enough work to know
what is going on, then we publish. I have been in situations where
others disagree with me so much that I just talk myself off of the
project. Not common, but it has happened. I consider these occasions
personal failures.

As the director of a facility, I regularly am helping people with
things about which I know nothing. I have a very different approach
here. I will only comment on what I know, the imaging system and know
artifacts there of. When I know the cell biology, I help with that
also. Once I have explained what I see in the sample, I let the
others do what they will. I am uncomfortable doing more that stating
my reservations. I have been known to repeat experiments to do the
controls that I thought a researcher should have done. If I can
replicate the result of interest in a negative control, then I get
more pushy about my thoughts.

Just my $.02
David


_____________________

David Elliott Ph.D.
Assistant Professor - Department of Cell Biology and Anatomy
Director, Research Microscopy Core Service
University of Arizona College of Medicine
PO Box 245044
Tucson, AZ 85724

Voice: 520-626-7870
Fax: 520-626-2097


On Jan 8, 2010, at 9:46 AM, TindallR-at-missouri.edu wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} The list has been a little slow lately, so here's a topic for
} potential discussion:
}
} On occasion we look at things in our scopes that have no good basis
} for reference---no previous publications, no other EM images to
} compare with---you get the picture. My method has been to take
} representative images of what is there, even if the images have a
} wide variety of things in them that don't resemble each other or
} what the sample supposedly "should" look like.
}
} It's that "should" that is the problem. It sometimes happens when
} we send these images to the clients that they grab onto whatever
} looks like what they want to see, pretty much ignore anything else,
} then starting making assertions about the images that go waaaay
} beyond what the image can support and want to plug all that into a
} publication. (I've had people get all Eureka! about the
} "champagning" artifact on a negatively-stained prep, for example.)
}
}
} So the question is, if the EM operator has a reasonable suspicion,
} but not a certainty, that an image is showing artifact or something
} that is not really the part of the sample the researcher wants to
} see, how should that be handled? Should we send the images along
} with our caveats and risk having them having them published with
} interpretations that go beyond the data and may just be dead wrong?
} Or should we self-censor and not send these images?
}
} Remember, I'm not talking about things that we KNOW are artifact or
} garbage. That's a clear call. I'm talking about imaging things
} that may not have been seen before, and nobody really knows what
} they look like (but they think they do), thereby making it difficult
} to separate artifact from real data. What we do now is send the
} images with our comments and hope that the client isn't so desperate
} for a publication that they ignore our cautions. We are virtually
} never listed as co-authors so that's not really an issue, but
} still.....I like clean science.
}
} How do other members of the Collective handle these cases?
}
} Cheers and stay warm!
}
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
}
}
}
} ==============================Original
} Headers==============================
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} by
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} Fri, 8 Jan 2010
} 13, 30 -- 10:43:46 -0600
} 13, 30 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu}
} 13, 30 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
} 13, 30 -- Date: Fri, 8 Jan 2010 10:43:45 -0600
} 13, 30 -- Subject: EM imaging: Self-censorship?
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From: bkang-at-ufl.edu
Date: Fri, 8 Jan 2010 12:01:07 -0600
Subject: [Microscopy] EM imaging: Self-censorship?

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

Hello,

I am in the same situation as Dr. Elliott. I work as a faculty at the microbiology and cell science department as well as a director of an EM lab at the core facility.

Basically, I do not stick only to electron microscopy for deriving conclusions. I reevaluate what we have found from electron microscopy by means of light microscopy, cell fractionation, or mutant characterization.

For service projects, I give my clients my comments and ask what other evidence they have to support their claim. For service projects in which we do not participate as a coauthor, it is their responsibility if clients go against opinions from an expert like you. I keep records of discussion (usually by e-mail) in case they blame me.

Hope this helps.

Thanks.

Byung-Ho Kang, Ph.D.
Assistant Professor, Microbiology and Cell Science
Director, Electron Microscopy and Bioimaging Lab, Interdisciplinary Center for Biotechnology Research
University of Florida Gainesville, FL 32611
Tel: 352-846-0952
Fax: 352-392-5922
http://microcell.ufl.edu/personnel/faculty/Kang.shtml

On Jan 8, 2010, at 12:14 PM, Elliott-at-arizona.edu wrote:

}
}
}
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}
} Good question Randy
}
} For my part, I have two hats. I am a faculty researcher and a person
} who directs a service facility.
}
} As a faculty member I am a bit pushy about things I know about. I
} know exactly what you are saying about people latching on to something
} they like and ignoring the rest. I push rather hard for a more
} neutral reading of the data. When we have done enough work to know
} what is going on, then we publish. I have been in situations where
} others disagree with me so much that I just talk myself off of the
} project. Not common, but it has happened. I consider these occasions
} personal failures.
}
} As the director of a facility, I regularly am helping people with
} things about which I know nothing. I have a very different approach
} here. I will only comment on what I know, the imaging system and know
} artifacts there of. When I know the cell biology, I help with that
} also. Once I have explained what I see in the sample, I let the
} others do what they will. I am uncomfortable doing more that stating
} my reservations. I have been known to repeat experiments to do the
} controls that I thought a researcher should have done. If I can
} replicate the result of interest in a negative control, then I get
} more pushy about my thoughts.
}
} Just my $.02
} David
}
}
} _____________________
}
} David Elliott Ph.D.
} Assistant Professor - Department of Cell Biology and Anatomy
} Director, Research Microscopy Core Service
} University of Arizona College of Medicine
} PO Box 245044
} Tucson, AZ 85724
}
} Voice: 520-626-7870
} Fax: 520-626-2097
}
}
} On Jan 8, 2010, at 9:46 AM, TindallR-at-missouri.edu wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} } America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } The list has been a little slow lately, so here's a topic for
} } potential discussion:
} }
} } On occasion we look at things in our scopes that have no good basis
} } for reference---no previous publications, no other EM images to
} } compare with---you get the picture. My method has been to take
} } representative images of what is there, even if the images have a
} } wide variety of things in them that don't resemble each other or
} } what the sample supposedly "should" look like.
} }
} } It's that "should" that is the problem. It sometimes happens when
} } we send these images to the clients that they grab onto whatever
} } looks like what they want to see, pretty much ignore anything else,
} } then starting making assertions about the images that go waaaay
} } beyond what the image can support and want to plug all that into a
} } publication. (I've had people get all Eureka! about the
} } "champagning" artifact on a negatively-stained prep, for example.)
} }
} }
} } So the question is, if the EM operator has a reasonable suspicion,
} } but not a certainty, that an image is showing artifact or something
} } that is not really the part of the sample the researcher wants to
} } see, how should that be handled? Should we send the images along
} } with our caveats and risk having them having them published with
} } interpretations that go beyond the data and may just be dead wrong?
} } Or should we self-censor and not send these images?
} }
} } Remember, I'm not talking about things that we KNOW are artifact or
} } garbage. That's a clear call. I'm talking about imaging things
} } that may not have been seen before, and nobody really knows what
} } they look like (but they think they do), thereby making it difficult
} } to separate artifact from real data. What we do now is send the
} } images with our comments and hope that the client isn't so desperate
} } for a publication that they ignore our cautions. We are virtually
} } never listed as co-authors so that's not really an issue, but
} } still.....I like clean science.
} }
} } How do other members of the Collective handle these cases?
} }
} } Cheers and stay warm!
} }
} } Randy
} }
} } Randy Tindall
} } Senior EM Specialist
} } Electron Microscopy Core Facility---We Do Small Well!
} } W125 Veterinary Medicine
} } University of Missouri
} } Columbia, MO 65211
} } Tel: (573) 882-8304
} } Fax: (573) 884-2227
} } Email: tindallr-at-missouri.edu
} } Web: http://www.emc.missouri.edu
} } On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} } Sons of Norway: http://www.sofn.com
} }
} }
} }
} }
} } ==============================Original
} } Headers==============================
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} } 13, 30 -- Date: Fri, 8 Jan 2010 10:43:45 -0600
} } 13, 30 -- Subject: EM imaging: Self-censorship?
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} } 13, 30 -- Thread-Index: AcqQgb56tpTajBGETxiqxqY+yQSBPQ==
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}









==============================Original Headers==============================
17, 25 -- From bkang-at-ufl.edu Fri Jan 8 12:01:07 2010
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From: marie.cantino-at-uconn.edu
Date: Fri, 8 Jan 2010 12:04:45 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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

I think we always need to discuss potential artifacts with our users
and suggest controls where these are feasible. This is particularly
true for assisted projects, where users are coming to us both for our
equipment and our expertise. Having done that, I am not interested in
getting into a protracted battle if the user still wants to proceed
with publication (and assuming I am not a coauthor).

Where we sometimes run into problems is when the contact is a student
who may not have the expertise or inclination to convey these
reservations to the advisor (who will be a coauthor and is usually
footing the bill). In such cases I think it's prudent to include any
comments and caveats in an e-mail copied to the advisor.

Marie

On Jan 8, 2010, at 11:50 AM, TindallR-at-missouri.edu wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} The list has been a little slow lately, so here's a topic for
} potential discussion:
}
} On occasion we look at things in our scopes that have no good basis
} for reference---no previous publications, no other EM images to
} compare with---you get the picture. My method has been to take
} representative images of what is there, even if the images have a
} wide variety of things in them that don't resemble each other or
} what the sample supposedly "should" look like.
}
} It's that "should" that is the problem. It sometimes happens when
} we send these images to the clients that they grab onto whatever
} looks like what they want to see, pretty much ignore anything else,
} then starting making assertions about the images that go waaaay
} beyond what the image can support and want to plug all that into a
} publication. (I've had people get all Eureka! about the
} "champagning" artifact on a negatively-stained prep, for example.)
}
}
} So the question is, if the EM operator has a reasonable suspicion,
} but not a certainty, that an image is showing artifact or something
} that is not really the part of the sample the researcher wants to
} see, how should that be handled? Should we send the images along
} with our caveats and risk having them having them published with
} interpretations that go beyond the data and may just be dead wrong?
} Or should we self-censor and not send these images?
}
} Remember, I'm not talking about things that we KNOW are artifact or
} garbage. That's a clear call. I'm talking about imaging things
} that may not have been seen before, and nobody really knows what
} they look like (but they think they do), thereby making it difficult
} to separate artifact from real data. What we do now is send the
} images with our comments and hope that the client isn't so desperate
} for a publication that they ignore our cautions. We are virtually
} never listed as co-authors so that's not really an issue, but
} still.....I like clean science.
}
} How do other members of the Collective handle these cases?
}
} Cheers and stay warm!
}
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
}
}
}
} ==============================Original
} Headers==============================
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} 13, 30 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu}
} 13, 30 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
} 13, 30 -- Date: Fri, 8 Jan 2010 10:43:45 -0600
} 13, 30 -- Subject: EM imaging: Self-censorship?
} 13, 30 -- Thread-Topic: EM imaging: Self-censorship?
} 13, 30 -- Thread-Index: AcqQgb56tpTajBGETxiqxqY+yQSBPQ==
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Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369


==============================Original Headers==============================
7, 21 -- From marie.cantino-at-uconn.edu Fri Jan 8 12:04:45 2010
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From: oshel1pe-at-cmich.edu
Date: Fri, 8 Jan 2010 12:16:08 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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

Randy,

First, the clients are going to over-interpret anyway, no matter how
well known or unknown the samples are, no matter what you say.
Sometimes one "gets it", but ...
And "clean science"? What's that? Especially in biology. It's all a
mess. Remember the main corollary to Heisenberg's Uncertainty
Principle: in any experiment, regardless of the results, you can
never know what really happened.
This goes for imaging things in the microscope, too. E.g., the size
of mammalian red blood cells depends on how they were prepared and
what microscopy was used to image them (the literature search is left
as an exercise for the reader). So how big is an RBC? It depends.
Before descending completely into gloom and despair, though, remember
we're *always* making judgements about how things should (or do)
look. So, if the specimens are new, we look more and use different
methods to look at them. If they're still biconcave discs in light
microscopy blood smears, DIC/phase, AFM, SEM, etc., RBCs probably
really are biconcave discs.
The problem isn't so much "how to interpet this new thing?" as "how
to interpet this new thing that I've only looked at one way?"

So, I'd send the images with the best interpetation and all the
caveats, foremost of which is "this needs more study and I suggest
these different ways of preparing and imaging." With the default
opinion that the whatzit is an artifact. ("Null hypothesis" if that
reads better.)

But I definitely would not self-censor. First, it's not really your
(our) data, it's the client's, and second, they may have literature
that refers to the whatzit or know someone you don't that has seen
the thing.

If the client is so desperate for publications that they ignore
cautions and caveats, then they're going to publish garbage even if
you only give them good, clean images. Just make *sure* you're not a
co-author, and maybe request that you're not in the acknowledgements.

Good luck. Keep the vacuum inside the column.

Phil

} The list has been a little slow lately, so here's a topic for
} potential discussion:
}
} On occasion we look at things in our scopes that have no good basis
} for reference---no previous publications, no other EM images to
} compare with---you get the picture. My method has been to take
} representative images of what is there, even if the images have a
} wide variety of things in them that don't resemble each other or
} what the sample supposedly "should" look like.
}
} It's that "should" that is the problem. It sometimes happens when
} we send these images to the clients that they grab onto whatever
} looks like what they want to see, pretty much ignore anything else,
} then starting making assertions about the images that go waaaay
} beyond what the image can support and want to plug all that into a
} publication. (I've had people get all Eureka! about the
} "champagning" artifact on a negatively-stained prep, for example.)
}
}
} So the question is, if the EM operator has a reasonable suspicion,
} but not a certainty, that an image is showing artifact or something
} that is not really the part of the sample the researcher wants to
} see, how should that be handled? Should we send the images along
} with our caveats and risk having them having them published with
} interpretations that go beyond the data and may just be dead wrong?
} Or should we self-censor and not send these images?
}
} Remember, I'm not talking about things that we KNOW are artifact or
} garbage. That's a clear call. I'm talking about imaging things
} that may not have been seen before, and nobody really knows what
} they look like (but they think they do), thereby making it difficult
} to separate artifact from real data. What we do now is send the
} images with our comments and hope that the client isn't so desperate
} for a publication that they ignore our cautions. We are virtually
} never listed as co-authors so that's not really an issue, but
} still.....I like clean science.
}
} How do other members of the Collective handle these cases?
}
} Cheers and stay warm!
}
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar:
} http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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From: thoward-at-unm.edu
Date: Fri, 8 Jan 2010 12:19:04 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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

One thing I've started to do is make notes directly on
certain images - the ones that I know will get someone all
excited over nothing (e.g., champagning in negative
stains, a recent cell culture I was given that had massive
Mycoplasma infection, etc.). I provide the original
digital images, then a copy of the suspect image with a
text layer pointing to the "problem", stating what it is &
why it isn't Nobel prize-worthy. Somehow seeing that info
right on the image gets the message across to some people
when a plain old text document accompanying the data disc
doesn't make a dent.

Great discussion topic, BTW.

Tamara

On Fri, 8 Jan 2010 10:44:58 -0600
TindallR-at-missouri.edu wrote:
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy
} Society of America
} To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
} On-Line Help
} http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} The list has been a little slow lately, so here's a
} topic for potential discussion:
}
} On occasion we look at things in our scopes that have no
} good basis for reference---no previous publications, no
} other EM images to compare with---you get the picture.
} My method has been to take representative images of what
} is there, even if the images have a wide variety of
} things in them that don't resemble each other or what the
} sample supposedly "should" look like.
}
} It's that "should" that is the problem. It sometimes
} happens when we send these images to the clients that
} they grab onto whatever looks like what they want to see,
} pretty much ignore anything else, then starting making
} assertions about the images that go waaaay beyond what
} the image can support and want to plug all that into a
} publication. (I've had people get all Eureka! about the
} "champagning" artifact on a negatively-stained prep, for
} example.)
}
}
} So the question is, if the EM operator has a reasonable
} suspicion, but not a certainty, that an image is showing
} artifact or something that is not really the part of the
} sample the researcher wants to see, how should that be
} handled? Should we send the images along with our
} caveats and risk having them having them published with
} interpretations that go beyond the data and may just be
} dead wrong? Or should we self-censor and not send these
} images?
}
} Remember, I'm not talking about things that we KNOW are
} artifact or garbage. That's a clear call. I'm talking
} about imaging things that may not have been seen before,
} and nobody really knows what they look like (but they
} think they do), thereby making it difficult to separate
} artifact from real data. What we do now is send the
} images with our comments and hope that the client isn't
} so desperate for a publication that they ignore our
} cautions. We are virtually never listed as co-authors so
} that's not really an issue, but still.....I like clean
} science.
}
} How do other members of the Collective handle these
} cases?
}
} Cheers and stay warm!
}
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar:
} http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
}
}
}
} ==============================Original
} Headers==============================
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} 2010
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} {TindallR-at-missouri.edu}
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} {microscopy-at-microscopy.com}
} 13, 30 -- Date: Fri, 8 Jan 2010 10:43:45 -0600
} 13, 30 -- Subject: EM imaging: Self-censorship?
} 13, 30 -- Thread-Topic: EM imaging: Self-censorship?
} 13, 30 -- Thread-Index: AcqQgb56tpTajBGETxiqxqY+yQSBPQ==
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***************************
Tamara Howard
Cell Biology & Physiology
UNM-HSC
Albuquerque, NM
***************************


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From: dcromey-at-email.arizona.edu
Date: Fri, 8 Jan 2010 12:39:56 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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

So what is our responsibility as scientists when someone (in our opinion)
crosses the line from an oddball interpretation of the data to an erroneous
and/or fraudulent interpretation? A previous facility manager here confided
in me that they no longer did service work for a particular faculty member
because of the way the faculty member had (in the manager's opinion) twisted
the data. Unfortunately the manager (who was not a faculty member) did not
feel as if there was an option for calling this behavior into question.
Whistleblowing can and/has historically left the whistleblower scarred or
unemployed. FWIW, our campus now has an anonymous phone number for
reporting financial and/or research fraud, but I have my doubts about how
well known it is on campus.

I've written guidelines about digital image manipulation ethics, but as
others have pointed out that it's easy to be outside of your area of
technical expertise when doing service work. Supposedly peer-review of
publications should weed out wacky interpretations, but we all know of odd
research findings that have been superseded by better research, or have seen
the Journals have to retract a paper because questions were raised about the
data (and further review by an embarrassed senior author who was not able to
locate the original data). A recent paper I read studied citations of the
articles that were involved in Office of Research Integrity findings. These
were cases where falsification/fabrication/plagiarism in the articles was
established. Of the articles written by others citing these "bad" papers,
only 5% of the citations referenced the fraudulent articles in a negative
light, the rest were considered positive. The blame goes a lot of places,
but don't some of us have a responsibility to try to reduce the amount of
"chaff" in the research literature? I'm not expecting a definitive answer,
just tossing this out there as a rhetorical question.

I once spent a good deal of time explaining to a student that the
colocalization they were seeing in confocal images (red image staining +
green image staining = yellow pixels in the overlay image, for you TEM
folks) was an artifact. The student, who was not a microscopist, could
never really explain the technical aspects of the problem to the PI and
ultimately I had to write a short essay (with illustrations) to explain the
physics to the PI. The lab really wanted the two items they were
immuno-staining to colocalize, but the confocal (configured correctly)
didn't show that.

Thanks for your good ideas.
Doug

^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Douglas W. Cromey, M.S. - Assistant Scientific Investigator
Dept. of Cell Biology & Anatomy, University of Arizona
1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA

office: AHSC 4212 email: Cromey-at-Arizona.edu
voice: 520-626-2824 fax: 520-626-2097

http://swehsc.pharmacy.arizona.edu/exppath/
Home of: "Microscopy and Imaging Resources on the WWW"

-----Original Message-----
X-from: marie.cantino-at-uconn.edu [mailto:marie.cantino-at-uconn.edu]
Sent: Friday, January 08, 2010 11:05 AM
To: dcromey-at-email.arizona.edu

I think we always need to discuss potential artifacts with our users
and suggest controls where these are feasible. This is particularly
true for assisted projects, where users are coming to us both for our
equipment and our expertise. Having done that, I am not interested in
getting into a protracted battle if the user still wants to proceed
with publication (and assuming I am not a coauthor).

Where we sometimes run into problems is when the contact is a student
who may not have the expertise or inclination to convey these
reservations to the advisor (who will be a coauthor and is usually
footing the bill). In such cases I think it's prudent to include any
comments and caveats in an e-mail copied to the advisor.

Marie

On Jan 8, 2010, at 11:50 AM, TindallR-at-missouri.edu wrote:

}
}
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}
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} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
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}
----------------------------------------------------------------------------
}
} The list has been a little slow lately, so here's a topic for
} potential discussion:
}
} On occasion we look at things in our scopes that have no good basis
} for reference---no previous publications, no other EM images to
} compare with---you get the picture. My method has been to take
} representative images of what is there, even if the images have a
} wide variety of things in them that don't resemble each other or
} what the sample supposedly "should" look like.
}
} It's that "should" that is the problem. It sometimes happens when
} we send these images to the clients that they grab onto whatever
} looks like what they want to see, pretty much ignore anything else,
} then starting making assertions about the images that go waaaay
} beyond what the image can support and want to plug all that into a
} publication. (I've had people get all Eureka! about the
} "champagning" artifact on a negatively-stained prep, for example.)
}
}
} So the question is, if the EM operator has a reasonable suspicion,
} but not a certainty, that an image is showing artifact or something
} that is not really the part of the sample the researcher wants to
} see, how should that be handled? Should we send the images along
} with our caveats and risk having them having them published with
} interpretations that go beyond the data and may just be dead wrong?
} Or should we self-censor and not send these images?
}
} Remember, I'm not talking about things that we KNOW are artifact or
} garbage. That's a clear call. I'm talking about imaging things
} that may not have been seen before, and nobody really knows what
} they look like (but they think they do), thereby making it difficult
} to separate artifact from real data. What we do now is send the
} images with our comments and hope that the client isn't so desperate
} for a publication that they ignore our cautions. We are virtually
} never listed as co-authors so that's not really an issue, but
} still.....I like clean science.
}
} How do other members of the Collective handle these cases?
}
} Cheers and stay warm!
}
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar:
http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week
&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
}
}
}
} ==============================Original
} Headers==============================
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} 13, 30 -- Subject: EM imaging: Self-censorship?
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Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369


==============================Original Headers==============================
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From: mcauliff-at-umdnj.edu
Date: Fri, 8 Jan 2010 12:45:30 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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

I am opposed to self-censorship which is saying (IMO) "I know more about
the project than the investigator therefore" .......
By all means do include caveats about sampling error, artifacts,
etc.but if someone chooses to "over interpret" that is there problem.

Geoff


TindallR-at-missouri.edu wrote:
} ----------------------------------------------------------------------------
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} The list has been a little slow lately, so here's a topic for potential discussion:
}
} On occasion we look at things in our scopes that have no good basis for reference---no previous publications, no other EM images to compare with---you get the picture. My method has been to take representative images of what is there, even if the images have a wide variety of things in them that don't resemble each other or what the sample supposedly "should" look like.
}
} It's that "should" that is the problem. It sometimes happens when we send these images to the clients that they grab onto whatever looks like what they want to see, pretty much ignore anything else, then starting making assertions about the images that go waaaay beyond what the image can support and want to plug all that into a publication. (I've had people get all Eureka! about the "champagning" artifact on a negatively-stained prep, for example.)
}
}
} So the question is, if the EM operator has a reasonable suspicion, but not a certainty, that an image is showing artifact or something that is not really the part of the sample the researcher wants to see, how should that be handled? Should we send the images along with our caveats and risk having them having them published with interpretations that go beyond the data and may just be dead wrong? Or should we self-censor and not send these images?
}
} Remember, I'm not talking about things that we KNOW are artifact or garbage. That's a clear call. I'm talking about imaging things that may not have been seen before, and nobody really knows what they look like (but they think they do), thereby making it difficult to separate artifact from real data. What we do now is send the images with our comments and hope that the client isn't so desperate for a publication that they ignore our cautions. We are virtually never listed as co-authors so that's not really an issue, but still.....I like clean science.
}
} How do other members of the Collective handle these cases?
}
} Cheers and stay warm!
}
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
}
}
}
} ==============================Original Headers==============================
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
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From: Frank_Karl-at-lincolnelectric.com
Date: Fri, 8 Jan 2010 13:40:16 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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

It isn't just biology. I examine broken metal rectangles, (called CVNs)
first to find the failure origin and then the cause of the failure. Like
many others, I do this for clients and I lose control of the image very
quickly.

In those cases where there appears to be an initiation point/origin but no
cause, I routine include that information in the file title. The notation
looks like "sample12344-56 suspected-origin-xxxmag" as compared to
"sample12344-56-origin-xxxmag". Many times material which could cause a
failure is observed near, but not at the suspected origin. Those are
labeled "sample12344-56-slag near suspected origin-xxxmag".

So what am I trying to say? I suspect the solution to releasing images,
which are owned by the client but which you have a stake in (you took it
after all) is to add limiting verbiage to the filename. I suspect
something like "-poss-artifact-" "--unexpt structure-" added to the
filename will at least have the client calling for additional information.

As it was mentioned, if the client makes outrageous claims, well you have
the images with the concern expressed in the filename...

Thank Heaven, I'm not limited to 12 characters in a file name.........

Frank

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From: dsherman-at-purdue.edu
Date: Fri, 8 Jan 2010 13:54:39 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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


Randy,
I have had similar concerns through out my research career. They go
back to the early days of Photoshop and the investigator who wanted to
alter intensities of bands in gels because he knew those extra bands
were just "mistakes".
We provide written reports with all service projects that contain all
sample prep info, summary of results, our observations, and any
explanations, suggestions,or concerns we have.
These are given to the students but also sent to the PI. Our
responsibility ends there unless we are asked for further input. The
reports are for our internal records as well as to help the researchers.
I also refuse co-authorships unless I have the opportunity to edit the
manuscript and agree with the conclusions.

Debby
Debby Sherman, Director
Life Science Microscopy Facility
Purdue University

Sent from my iPhone

On Jan 8, 2010, at 8:45 AM, "TindallR-at-missouri.edu" {TindallR-at-missouri.edu
} wrote:

}
}
}
} ---
} ---
} ----------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ---
} ---
} ----------------------------------------------------------------------
}
} The list has been a little slow lately, so here's a topic for
} potential discussion:
}
} On occasion we look at things in our scopes that have no good basis
} for reference---no previous publications, no other EM images to
} compare with---you get the picture. My method has been to take
} representative images of what is there, even if the images have a
} wide variety of things in them that don't resemble each other or
} what the sample supposedly "should" look like.
}
} It's that "should" that is the problem. It sometimes happens when
} we send these images to the clients that they grab onto whatever
} looks like what they want to see, pretty much ignore anything else,
} then starting making assertions about the images that go waaaay
} beyond what the image can support and want to plug all that into a
} publication. (I've had people get all Eureka! about the
} "champagning" artifact on a negatively-stained prep, for example.)
}
}
} So the question is, if the EM operator has a reasonable suspicion,
} but not a certainty, that an image is showing artifact or something
} that is not really the part of the sample the researcher wants to
} see, how should that be handled? Should we send the images along
} with our caveats and risk having them having them published with
} interpretations that go beyond the data and may just be dead wrong?
} Or should we self-censor and not send these images?
}
} Remember, I'm not talking about things that we KNOW are artifact or
} garbage. That's a clear call. I'm talking about imaging things
} that may not have been seen before, and nobody really knows what
} they look like (but they think they do), thereby making it difficult
} to separate artifact from real data. What we do now is send the
} images with our comments and hope that the client isn't so desperate
} for a publication that they ignore our cautions. We are virtually
} never listed as co-authors so that's not really an issue, but
} still.....I like clean science.
}
} How do other members of the Collective handle these cases?
}
} Cheers and stay warm!
}
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
}
}
}
} ==============================Original
} Headers==============================
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} 13, 30 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
} 13, 30 -- Date: Fri, 8 Jan 2010 10:43:45 -0600
} 13, 30 -- Subject: EM imaging: Self-censorship?
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From: lcgould-at-med.cornell.edu
Date: Fri, 8 Jan 2010 14:54:42 -0600
Subject: [Microscopy] EMI systems-summary

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Hi All-
I'd like to thank everyone who responded to my earlier inquiry.
I've had responses on-list, off-list and by telephone. Some people
contacted me with suggestions for hunting down the source of our
spike, others simply to commiserate, and others to offer comments
about the systems they have in their facilities.
It seems that most of the people who have systems to cancel EMI are
happy with them, regardless of the manufacturer. They all seem to do
the job. That said, the decision might boil down to cost but the
estimates I got were all in the same ballpark, so a couple of
thousand one way or another won't be my deciding factor.
I did decide on a full room system rather than a cage around the
column or the microscope for both esthetics and ease of access to the
microscope.
The vendors may be saddened to hear this, but the systems just don't
seem to be that different from one another when you get down to the
user's perspective.
So, get your estimates, talk to the vendors' reps. It may boil down
to using the company that has engineers nearest you geographically,
or with whom you establish an easy rapport.
They did the rocket science, you don't have to.

Lee
--
Lee Cohen-Gould, M.S.
Sr. Staff Associate in Biochemistry and
Cell & Developmental Biology
Director, Electron Microscopy & Histology
and Optical Microscopy Core Facilities
Weill Cornell Medical College

voice (212)746-6146
fax (212)746-8175
http://www.med.cornell.edu/research/rea_sup/
http://www.cornellcelldevbiology.org
http://www.cornellbiochem.org

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From: naomi_mccallum-at-health.qld.gov.au
Date: Fri, 8 Jan 2010 17:44:34 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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Great discussion, thank you.

Please spare a thought for those working in the field of Diagnostic Pathology!

regards
Naomi


Naomi McCallum
Supervising Scientist, EM Unit
Pathology Queensland



} } } {TindallR-at-missouri.edu} 9/01/2010 2:53 am } } }



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The list has been a little slow lately, so here's a topic for potential discussion:

On occasion we look at things in our scopes that have no good basis for reference---no previous publications, no other EM images to compare with---you get the picture. My method has been to take representative images of what is there, even if the images have a wide variety of things in them that don't resemble each other or what the sample supposedly "should" look like.

It's that "should" that is the problem. It sometimes happens when we send these images to the clients that they grab onto whatever looks like what they want to see, pretty much ignore anything else, then starting making assertions about the images that go waaaay beyond what the image can support and want to plug all that into a publication. (I've had people get all Eureka! about the "champagning" artifact on a negatively-stained prep, for example.)


So the question is, if the EM operator has a reasonable suspicion, but not a certainty, that an image is showing artifact or something that is not really the part of the sample the researcher wants to see, how should that be handled? Should we send the images along with our caveats and risk having them having them published with interpretations that go beyond the data and may just be dead wrong? Or should we self-censor and not send these images?

Remember, I'm not talking about things that we KNOW are artifact or garbage. That's a clear call. I'm talking about imaging things that may not have been seen before, and nobody really knows what they look like (but they think they do), thereby making it difficult to separate artifact from real data. What we do now is send the images with our comments and hope that the client isn't so desperate for a publication that they ignore our cautions. We are virtually never listed as co-authors so that's not really an issue, but still.....I like clean science.

How do other members of the Collective handle these cases?

Cheers and stay warm!

Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




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From: arnec-at-bio.umass.edu
Date: Sat, 9 Jan 2010 09:10:17 -0600
Subject: [Microscopy] viaWWW: Histology stains for the retina

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Email: arnec-at-bio.umass.edu
Name: Arne

Title-Subject: [Filtered] Histology stains for the retina

Question: Dear Listeners,

I'm putting together a lab-based presentation for an undergraduate
biology class. The presentation will focus on the retina. I have some
PFA fixed retinal tissue sections, and I'm seeking advice for
vibrant, informative stains. I'm not familiar with many of the
standard histological staining protocols, as I deal more with IF, but
the lab is not set up for fluorescent microscopy. We have these on
hand:

Acriflavin
Basic Fuchsin
Carmine Aluma Lake
Crystal Violet
Eosin
Fast Green FCF
Giesma
Malachite Green
Meyer's Hematoxylin
Orcein
Sudan Black B
Toluidine Blue O
Trypan Blue

Does anybody have any favorite stains/protocols for retinal tissue?

Thank you,
Arne

Login Host: 159.189.36.78
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From: gary-at-gaugler.com
Date: Sat, 9 Jan 2010 10:23:00 -0600
Subject: [Microscopy] EM imaging: Self-censorship?

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

At 11:41 AM 1/8/2010, you wrote:



[snip]


} So what am I trying to say? I suspect the solution to releasing images,
} which are owned by the client but which you have a stake in (you took it
} after all) is to add limiting verbiage to the filename. I suspect
} something like "-poss-artifact-" "--unexpt structure-" added to the
} filename will at least have the client calling for additional information.

[snip]

How about a different question relative to this topic, among others?

If a microscopist takes a pix for a client, how is the copyright
handled, if at all? By law, the microscopist owns the copyright.
Charging for the pix does not transfer the copyright.

I'm sure that the pix are never registered. How does this work
in practice? Is there a contract or Ts&Cs that say that the
copyright passes to the client?

Just wondering.

gary g.


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From: PhillipsT-at-missouri.edu
Date: Sat, 9 Jan 2010 12:05:12 -0600
Subject: [Microscopy] EM imaging: Self-censorship?

Contents Retrieved from Microscopy Listserver Archives
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Any image published in a journal is copyrighted and in almost all cases that copyright belongs to the journal. One generally has to sign away the copyright when submitting a paper for publication.

But one doesn't have to publish or register a work for it to be copyrighted. Creation of the work is enough.

I am not sure I buy your argument that the copyright belongs to the microscopist who took the image. I am not a lawyer but currently sitting on a committee re-writing our University's intellectual property rules. The lawyer on the committee recently stated that all who contribute to the creation of a work have equal rights. So if anyone in a lab prepared the tissue, fixed, embedded, sectioned, etc. they might have some claim. On the other hand, standard practice is the Principal Investigator "owns" the data in the sense he or she is responsible for maintaining it - others have additional rights and responsibilities but typical lab policy is that original data, whether it is photographic, gels, Western/Northern/Southern blots, or lab notebooks would remain in the laboratory where they were created. My students and technicians are not allowed to take the original copies of notebooks out of the lab but have the opportunity to take copies of all their work. My guess is that is a core technician claimed "ownership" rights to an image done on a strict for fee basis, they might have some rights but would be looking for a new job. The International Committee of Medical Journal Editorships (http://www.icmje.org/ethical_1author.html) states that "Authorship credit should be based on 1) substantial contributions to conception and design, acquisition of data, or analysis and interpretation of data; 2) drafting the article or revising it critically for important intellectual content; and 3) final approval of the version to be published. Authors should meet conditions 1, 2, and 3." Simply photographing an image isn't sufficient to claim a right to authorship and it shouldn't be enough to claim "ownership" of the interpretation.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com]
Sent: Saturday, January 09, 2010 10:24 AM
To: Phillips, Thomas E.

At 11:41 AM 1/8/2010, you wrote:



[snip]


} So what am I trying to say? I suspect the solution to releasing images,
} which are owned by the client but which you have a stake in (you took it
} after all) is to add limiting verbiage to the filename. I suspect
} something like "-poss-artifact-" "--unexpt structure-" added to the
} filename will at least have the client calling for additional information.

[snip]

How about a different question relative to this topic, among others?

If a microscopist takes a pix for a client, how is the copyright
handled, if at all? By law, the microscopist owns the copyright.
Charging for the pix does not transfer the copyright.

I'm sure that the pix are never registered. How does this work
in practice? Is there a contract or Ts&Cs that say that the
copyright passes to the client?

Just wondering.

gary g.


==============================Original Headers==============================
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From: FMonson-at-wcupa.edu
Date: Sun, 10 Jan 2010 18:40:54 -0600
Subject: [Microscopy] EM imaging: Self-censorship?

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


________________________________________
X-from: PhillipsT-at-missouri.edu [PhillipsT-at-missouri.edu]
Sent: Saturday, January 09, 2010 1:14 PM
To: Monson, Frederick

Any image published in a journal is copyrighted and in almost all cases that copyright belongs to the journal. One generally has to sign away the copyright when submitting a paper for publication.

But one doesn't have to publish or register a work for it to be copyrighted. Creation of the work is enough.

I am not sure I buy your argument that the copyright belongs to the microscopist who took the image. I am not a lawyer but currently sitting on a committee re-writing our University's intellectual property rules. The lawyer on the committee recently stated that all who contribute to the creation of a work have equal rights. So if anyone in a lab prepared the tissue, fixed, embedded, sectioned, etc. they might have some claim. On the other hand, standard practice is the Principal Investigator "owns" the data in the sense he or she is responsible for maintaining it - others have additional rights and responsibilities but typical lab policy is that original data, whether it is photographic, gels, Western/Northern/Southern blots, or lab notebooks would remain in the laboratory where they were created. My students and technicians are not allowed to take the original copies of notebooks out of the lab but have the opportunity to take copies of all their work. My guess is that is a!
core technician claimed "ownership" rights to an image done on a strict for fee basis, they might have some rights but would be looking for a new job. The International Committee of Medical Journal Editorships (http://www.icmje.org/ethical_1author.html) states that "Authorship credit should be based on 1) substantial contributions to conception and design, acquisition of data, or analysis and interpretation of data; 2) drafting the article or revising it critically for important intellectual content; and 3) final approval of the version to be published. Authors should meet conditions 1, 2, and 3." Simply photographing an image isn't sufficient to claim a right to authorship and it shouldn't be enough to claim "ownership" of the interpretation.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com]
Sent: Saturday, January 09, 2010 10:24 AM
To: Phillips, Thomas E.

At 11:41 AM 1/8/2010, you wrote:



[snip]


} So what am I trying to say? I suspect the solution to releasing images,
} which are owned by the client but which you have a stake in (you took it
} after all) is to add limiting verbiage to the filename. I suspect
} something like "-poss-artifact-" "--unexpt structure-" added to the
} filename will at least have the client calling for additional information.

[snip]

How about a different question relative to this topic, among others?

If a microscopist takes a pix for a client, how is the copyright
handled, if at all? By law, the microscopist owns the copyright.
Charging for the pix does not transfer the copyright.

I'm sure that the pix are never registered. How does this work
in practice? Is there a contract or Ts&Cs that say that the
copyright passes to the client?

Just wondering.

gary g.


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26, 34 -- To: "gary-at-gaugler.com" {gary-at-gaugler.com} ,
26, 34 -- "microscopy-at-microscopy.com"
26, 34 -- {microscopy-at-microscopy.com}
26, 34 -- Date: Sat, 9 Jan 2010 12:05:09 -0600
26, 34 -- Subject: RE: [Microscopy] EM imaging: Self-censorship?
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From: gary-at-gaugler.com
Date: Sun, 10 Jan 2010 19:53:41 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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

You are mixing a bunch of variables here. Specimen prep,
documentation, notebooks, etc. are irrelevant with regards
to a photographic image. Copyright law specifies that the
person taking the image owns the copyright to that image.

Regarding publishing, yes, the venue requires transfer of
copyright. Hence, I do not publish with pix worth money.
For academic instances, I doubt that there is any argument
about the image ownership and corresponding transfer of
copyright.

} The International Committee of Medical Journal Editorships
} (http://www.icmje.org/ethical_1author.html) states that "Authorship
} credit should be based on 1) substantial contributions to conception
} and design, acquisition of data, or analysis and interpretation of
} data; 2) drafting the article or revising it critically for
} important intellectual content; and 3) final approval of the version
} to be published. Authors should meet conditions 1, 2, and 3." Simply
} photographing an image isn't sufficient to claim a right to
} authorship and it shouldn't be enough to claim "ownership" of the
} interpretation.

Again, this topic has no connection to the image. These are separate issues.
Authorship of the article is not tied to who took any images. The only
tertiary issue is that if the image is submitted as a "Publication as
a Contribution."
This pertains to a periodical, serial or collection.

The transfer of the author's inherent/indigenous copyright either freely or
for a fee dismisses the original author's copyright and transfers
it to the second party. This seems simple enough. But the story does
not stop here. The issue of copyright registration remains a big deal.
I refer you to US Copyright Office Form VA. There is no provision for
specimen prep, et. al.

This form, plus deposit of representation of the works plus the deposit fee
REGISTERS the copyright to the copyright owner. This is a big deal. Huge.
If someone infringes on the original inherent copyright, the extent
of collection of damages is limited to actual loss. In normal practice,
that is trivial and difficult to prove. However, if a registered image
is infringed, that allows punitive damages to be added to any actual
loss--and the actual
loss is examined more closely.

Form VA allows for multiple authors. But the nature of Authorship for
what we are talking about is focused on "Photograph." Specimen prep, et. al.
are not at all at issue or relevant for Form VA and thus, formal registration.

Infringing on a registered image is a serious issue and can cost the
infringer big bucks...depending on the specifics of the infringement.
It could be $1K to $100K...or more. As long as a potential user
knows that the image is registered, they are much more careful
to a frontal charge against a registered image.

I am not a lawyer, but my registered images have been infringed.
All instances were settled amicably. This issue is infrequent.

I would encourage more discussion about this topic to help
those who have to deal with this issue either for publication,
ownership, or whatever.

Should I change the Subject entry from Self-censorship to copyrights?
I recall discussing this copyright topic on the list perhaps five
years ago. AFAK, the law is the same today as then...but there
are efforts underway to torpedo orphan works.

gary g.




At 04:43 PM 1/10/2010, you wrote:



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


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20, 20 -- Subject: Re: [Microscopy] EM imaging: Self-censorship?
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From: PhillipsT-at-missouri.edu
Date: Sun, 10 Jan 2010 20:05:35 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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

To say the individual who takes an EM or LM photo has more rights than the one who fixed, embedded and sectioned it or the one who designed the experiment or the one who wrote the grant that got it suggests we have different backgrounds and understanding of how science is done but I don't have interest, expertise or time to get into a discussion of arcane details of copyright. Two non-lawyers discussing it is not especially authoritative nor useful. I don't see this as an issue that will ever impact my career as a research scientist. But I don't accept your interpretation since it contains circular logic. I can't transfer copyright unless I own it.

Thomas E. Phillips, Ph.D.
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
Biological Sciences
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (voice)
573-882-0123 (fax)


-----Original Message-----
X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com]
Sent: Sunday, January 10, 2010 7:54 PM
To: Phillips, Thomas E.

You are mixing a bunch of variables here. Specimen prep,
documentation, notebooks, etc. are irrelevant with regards
to a photographic image. Copyright law specifies that the
person taking the image owns the copyright to that image.

Regarding publishing, yes, the venue requires transfer of
copyright. Hence, I do not publish with pix worth money.
For academic instances, I doubt that there is any argument
about the image ownership and corresponding transfer of
copyright.

} The International Committee of Medical Journal Editorships
} (http://www.icmje.org/ethical_1author.html) states that "Authorship
} credit should be based on 1) substantial contributions to conception
} and design, acquisition of data, or analysis and interpretation of
} data; 2) drafting the article or revising it critically for
} important intellectual content; and 3) final approval of the version
} to be published. Authors should meet conditions 1, 2, and 3." Simply
} photographing an image isn't sufficient to claim a right to
} authorship and it shouldn't be enough to claim "ownership" of the
} interpretation.

Again, this topic has no connection to the image. These are separate issues.
Authorship of the article is not tied to who took any images. The only
tertiary issue is that if the image is submitted as a "Publication as
a Contribution."
This pertains to a periodical, serial or collection.

The transfer of the author's inherent/indigenous copyright either freely or
for a fee dismisses the original author's copyright and transfers
it to the second party. This seems simple enough. But the story does
not stop here. The issue of copyright registration remains a big deal.
I refer you to US Copyright Office Form VA. There is no provision for
specimen prep, et. al.

This form, plus deposit of representation of the works plus the deposit fee
REGISTERS the copyright to the copyright owner. This is a big deal. Huge.
If someone infringes on the original inherent copyright, the extent
of collection of damages is limited to actual loss. In normal practice,
that is trivial and difficult to prove. However, if a registered image
is infringed, that allows punitive damages to be added to any actual
loss--and the actual
loss is examined more closely.

Form VA allows for multiple authors. But the nature of Authorship for
what we are talking about is focused on "Photograph." Specimen prep, et. al.
are not at all at issue or relevant for Form VA and thus, formal registration.

Infringing on a registered image is a serious issue and can cost the
infringer big bucks...depending on the specifics of the infringement.
It could be $1K to $100K...or more. As long as a potential user
knows that the image is registered, they are much more careful
to a frontal charge against a registered image.

I am not a lawyer, but my registered images have been infringed.
All instances were settled amicably. This issue is infrequent.

I would encourage more discussion about this topic to help
those who have to deal with this issue either for publication,
ownership, or whatever.

Should I change the Subject entry from Self-censorship to copyrights?
I recall discussing this copyright topic on the list perhaps five
years ago. AFAK, the law is the same today as then...but there
are efforts underway to torpedo orphan works.

gary g.




At 04:43 PM 1/10/2010, you wrote:



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


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20, 20 -- From: Gary Gaugler {gary-at-gaugler.com}
20, 20 -- Subject: Re: [Microscopy] EM imaging: Self-censorship?
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31, 40 -- Date: Sun, 10 Jan 2010 20:05:27 -0600
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From: gary-at-gaugler.com
Date: Sun, 10 Jan 2010 20:22:38 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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

Please do tell what is the circular logic about this.
I would like to understand your view(s) on this.

I agree that you nor anyone can transfer a copyright that they
do not own.

As far as copyrights are concerned, science is not in the "picture."

If you care to, read the law. Here is a good place to start:

http://www.copyright.gov/

In your situation, the issue is perhaps arcane if not moot. For others, it
is a big financial issue. It is not an all-consuming issue for me
but more of a CYA. Fortunately, if I can say so that being a pain to deal with
this, on just a few disturbing occasions it has paid off. My goal is not
to make money from infringements but rather to protect my IP for legal
licensing.

gary g.


At 06:06 PM 1/10/2010, you wrote:



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From: gary-at-gaugler.com
Date: Sun, 10 Jan 2010 20:29:19 -0600
Subject: [Microscopy] Re: EM imaging: Self-censorship?

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

On thinking about this, if the university, government or company
was astute, they would have required the employee being
highered to automatically assign their native copyright(s)
to the higher authority. In this case, the personal ownership of
the copyright is moot.

gary g.


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

Please do tell what is the circular logic about this.
I would like to understand your view(s) on this.

I agree that you nor anyone can transfer a copyright that they
do not own.

As far as copyrights are concerned, science is not in the "picture."

If you care to, read the law. Here is a good place to start:

http://www.copyright.gov/

In your situation, the issue is perhaps arcane if not moot. For others, it
is a big financial issue. It is not an all-consuming issue for me
but more of a CYA. Fortunately, if I can say so that being a pain to deal with
this, on just a few disturbing occasions it has paid off. My goal is not
to make money from infringements but rather to protect my IP for legal
licensing.

gary g.


At 06:06 PM 1/10/2010, you wrote:



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From: Frank_Karl-at-lincolnelectric.com
Date: Mon, 11 Jan 2010 06:09:49 -0600
Subject: [Microscopy] Re: FW: EM imaging: Self-censorship?

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


-----Original Message-----
X-from: Vince Crist [mailto:bvcrist-at-xpsdata.com]
Sent: Sunday, January 10, 2010 8:33 PM
To: Phillips, Thomas E.

Interesting conversation. With the disclaimer, I'm not a lawyer nor do I
play one on television, I wanted to mention that every company I worked for
required me to sign a document that any image or idea, even if developed at
home on my time, was company property. There were mechanism in place to
obtain ownership of ideas and images they did not want.

This discussion reminds me of the ones I use to read in the photo magazines
back in the darkroom days. Both professional and semi-professional were
worried about image theft. Anyone could copyright an image. It just took
money and a little time, both were in short supply. One solution was to
copyright a contact proof sheet which contained 36 small images.

In industry, I'm not interested in the financial aspect of images or who
gets credit. I'm more interested in misuse of "my" image and connection to
me and the company I work for. I worked with a person who presented images
in support of a theory on the incorporation of carbon black in stock. The
images were from another microscopist in the group. I couldn't see it in
the images and neither could anyone else except the presenter. The later
discredited work reflected badly on the microscopy group and of course on
the microscopist who had no control over the use of the images.


Frank

--
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replying to the message and deleting it from your computer.
Thank you,
The Lincoln Electric Company
**************************************************************


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From: TindallR-at-missouri.edu
Date: Mon, 11 Jan 2010 08:45:43 -0600
Subject: [Microscopy] Re: FW: EM imaging: Self-censorship?

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

Which pretty much brings us back to my original question, methinks.

Randy

-----Original Message-----
X-from: Frank_Karl-at-lincolnelectric.com [mailto:Frank_Karl-at-lincolnelectric.com]
Sent: Monday, January 11, 2010 6:11 AM
To: Tindall, Randy D.

Interesting conversation. With the disclaimer, I'm not a lawyer nor do I
play one on television, I wanted to mention that every company I worked for
required me to sign a document that any image or idea, even if developed at
home on my time, was company property. There were mechanism in place to
obtain ownership of ideas and images they did not want.

This discussion reminds me of the ones I use to read in the photo magazines
back in the darkroom days. Both professional and semi-professional were
worried about image theft. Anyone could copyright an image. It just took
money and a little time, both were in short supply. One solution was to
copyright a contact proof sheet which contained 36 small images.

In industry, I'm not interested in the financial aspect of images or who
gets credit. I'm more interested in misuse of "my" image and connection to
me and the company I work for. I worked with a person who presented images
in support of a theory on the incorporation of carbon black in stock. The
images were from another microscopist in the group. I couldn't see it in
the images and neither could anyone else except the presenter. The later
discredited work reflected badly on the microscopy group and of course on
the microscopist who had no control over the use of the images.


Frank

--
*************************************************************
Note:
The information contained in this message may be
privileged and confidential and protected from disclosure. If
the reader of this message is not the intended recipient, or
an employee or agent responsible for delivering this message
to the intended recipient, you are hereby notified that any
dissemination, distribution or copying of this communication
is strictly prohibited. If you have received this
communication in error, please notify us immediately by
replying to the message and deleting it from your computer.
Thank you,
The Lincoln Electric Company
**************************************************************


==============================Original Headers==============================
7, 22 -- From frank_karl-at-lincolnelectric.com Mon Jan 11 06:09:49 2010
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15, 33 -- From TindallR-at-missouri.edu Mon Jan 11 08:45:42 2010
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15, 33 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu}
15, 33 -- To: "Frank_Karl-at-lincolnelectric.com" {Frank_Karl-at-lincolnelectric.com}
15, 33 -- CC: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
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From: syrahman-at-uab.edu
Date: Mon, 11 Jan 2010 14:43:34 -0600
Subject: [Microscopy] EM imaging: Self-censorship?

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

I agree Dr Phillip's point that photographing an image isn't sufficient to claim a right to authorship and it shouldn't be enough to claim "ownership" of the interpretation. ICMJE criteria you mentioned are basic rules for those who involved in the project. Post-doctoral fellow of the concern project ( should be first Author) and PI ( Co author) are the main authority in making decision because post-doc is the one who starts project from the scratch, does literature search, make changes and presents every week under supervision of principle investigator. Any addition or subtraction is part of ongoing scientific process. Acknowledgment is another way to put the name for someone who can provide support but cannot claim it. According to ICMJE, criteria for acknowledgement is,



"All contributors who do not meet the criteria for authorship should be listed in an acknowledgments section. Examples of those who might be acknowledged include a person who provided purely technical help, writing assistance, or a department chair who provided only general support. Editors should ask corresponding authors to declare whether they had assistance with study design, data collection, data analysis, or manuscript preparation. If such assistance was available, the authors should disclose the identity of the individuals who provided this assistance and the entity that supported it in the published article. Financial and material support should also be acknowledged.

Groups of persons who have contributed materially to the paper but whose contributions do not justify authorship may be listed under such headings as "clinical investigators" or "participating investigators," and their function or contribution should be described-for example, "served as scientific advisors," "critically reviewed the study proposal," "collected data," or "provided and cared for study patients." Because readers may infer their endorsement of the data and conclusions, these persons must give written permission to be acknowledged".


I think ownership of picture or material is purely Post-doc and Principle investigator property. Any additional change during study comes under acknowledgement who can not claim for ownership for any picture or any material used for study.


Regards,



Syed Rahmanuddin, MD (MBBS)

Imaging Research Fellow

Department of Medicine

611A Tinsley Harrison Towers

1900 University Boulevard

Birmingham, AL 35294-0006

(205) 996-9003

syrahman-at-uab.edu











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Any image published in a journal is copyrighted and in almost all cases that copyright belongs to the journal. One generally has to sign away the copyright when submitting a paper for publication.



But one doesn't have to publish or register a work for it to be copyrighted. Creation of the work is enough.



I am not sure I buy your argument that the copyright belongs to the microscopist who took the image. I am not a lawyer but currently sitting on a committee re-writing our University's intellectual property rules. The lawyer on the committee recently stated that all who contribute to the creation of a work have equal rights. So if anyone in a lab prepared the tissue, fixed, embedded, sectioned, etc. they might have some claim. On the other hand, standard practice is the Principal Investigator "owns" the data in the sense he or she is responsible for maintaining it - others have additional rights and responsibilities but typical lab policy is that original data, whether it is photographic, gels, Western/Northern/Southern blots, or lab notebooks would remain in the laboratory where they were created. My students and technicians are not allowed to take the original copies of notebooks out of the lab but have the opportunity to take copies of all their work. My guess is that is a!

core technician claimed "ownership" rights to an image done on a strict for fee basis, they might have some rights but would be looking for a new job. The International Committee of Medical Journal Editorships (http://www.icmje.org/ethical_1author.html) states that "Authorship credit should be based on 1) substantial contributions to conception and design, acquisition of data, or analysis and interpretation of data; 2) drafting the article or revising it critically for important intellectual content; and 3) final approval of the version to be published. Authors should meet conditions 1, 2, and 3." Simply photographing an image isn't sufficient to claim a right to authorship and it shouldn't be enough to claim "ownership" of the interpretation.





Thomas E. Phillips, Ph.D

Professor of Biological Sciences

Director, Molecular Cytology Core

2 Tucker Hall

University of Missouri

Columbia, MO 65211-7400

573-882-4712 (office)

573-882-0123 (fax)

phillipst-at-missouri.edu



http://www.biology.missouri.edu/faculty/phillips.html

http://www.biotech.missouri.edu/mcc/





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

X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com]

Sent: Saturday, January 09, 2010 10:24 AM

To: Phillips, Thomas E.



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



At 11:41 AM 1/8/2010, you wrote:







[snip]





} So what am I trying to say? I suspect the solution to releasing images,

} which are owned by the client but which you have a stake in (you took it

} after all) is to add limiting verbiage to the filename. I suspect

} something like "-poss-artifact-" "--unexpt structure-" added to the

} filename will at least have the client calling for additional information.



[snip]



How about a different question relative to this topic, among others?



If a microscopist takes a pix for a client, how is the copyright

handled, if at all? By law, the microscopist owns the copyright.

Charging for the pix does not transfer the copyright.



I'm sure that the pix are never registered. How does this work

in practice? Is there a contract or Ts&Cs that say that the

copyright passes to the client?



Just wondering.



gary g.



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From: Michal.Jarnik-at-fccc.edu
Date: Mon, 11 Jan 2010 14:54:02 -0600
Subject: [Microscopy] Third party service contracts

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

Hi List,

I am looking for a little more economic way of taking care of our FEI
Tecnai 12. While I would be very hesitant to take our instrument off
the service contract completely, maybe I do not have pay k$20 a year
either. I am in Philadelphia area, any tips (on- or off-list) appreciated.

Thanks, Michal

--

Michal Jarnik, Ph.D.
Fox Chase Cancer Center
Philadelphia, PA


==============================Original Headers==============================
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From: TindallR-at-missouri.edu
Date: Mon, 11 Jan 2010 16:06:02 -0600
Subject: [Microscopy] Third party service contracts

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

Hi Michal,

I have no experience with third party service providers, although I know some people have been very happy with them. My purpose in replying is to caution you against going with "insurance company" service contracts. Bad news, IMHO. These involve a company that manages your service contracts and removes you a step from your service provider. They may cost less, but how much is not getting a permanent migraine worth?

I strongly suggest that you deal directly with whomever does your actual service----OEM or 3rd party.

Good luck,
Randy


Electron Microscopy Core Staff
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304 / 4777
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
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-----Original Message-----
X-from: Michal.Jarnik-at-fccc.edu [mailto:Michal.Jarnik-at-fccc.edu]
Sent: Monday, January 11, 2010 2:55 PM
To: Tindall, Randy D.

Hi List,

I am looking for a little more economic way of taking care of our FEI
Tecnai 12. While I would be very hesitant to take our instrument off
the service contract completely, maybe I do not have pay k$20 a year
either. I am in Philadelphia area, any tips (on- or off-list) appreciated.

Thanks, Michal

--

Michal Jarnik, Ph.D.
Fox Chase Cancer Center
Philadelphia, PA


==============================Original Headers==============================
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From: jsilvey-at-tegal.com
Date: Mon, 11 Jan 2010 17:46:07 -0600
Subject: [Microscopy] viaWWW: Epoxy fill material alternatives sought

Contents Retrieved from Microscopy Listserver Archives
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Email: jsilvey-at-tegal.com
Name: Judy Silvey

Organization: Tegal Corp.

Title-Subject: [Filtered] Epoxy

Message: Epoxy fill material alternatives sought



I am trying to characterize the deposition of polymer layers on the
sidewalls of deep trenches in silicon. The polymer layers are soft
and require polishing to capture the thickness and coverage of the
deep trenches. I have been using acrylic-based epoxies to fill the
trenches prior to polishing but these acrylic materials tend to
deform during the polishing process and subsequent exposure to the
beam of electrons in the SEM.



I am looking for an epoxy or other fill material that will not deform
when exposed to the polishing process or the SEM, and that does not
require a high temperature anneal to cure. A room
temperature-curable material is preferred if the cure time is less
than ~24hrs. I suspect that a material that has a hardness similar
to that of the surrounding silicon such a silicon dioxide or
alumina-based epoxy is best but this has not been proven (in contrast
to the acrylic epoxies that appear to be much softer.) I have
considered using a spin-on-glass but the reported cure temperatures
are too high, although a low temperature cure for an extended period
might in theory work. (has anyone tried this?)



Any ideas on good candidates for alternatives to the acrylic epoxies
for filling deep trenches (~50um) in silicon that will tolerate the
polishing process and the SEM exposure?




Login Host: 72.245.193.146
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From: r-holdford-at-ti.com
Date: Mon, 11 Jan 2010 20:15:41 -0600
Subject: [Microscopy] Re: viaWWW: Epoxy fill material alternatives sought

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

Judy: I like EpoxyBond 110 from Allied High Tech. It's two-part,
low-viscosity, heat-cure epoxy that I use routinely in semiconductor
work. I use when I prepare samples using the glass cover slip
technique, although I suppose one could spin it on if needed. I usually
cure for 10 minutes at 120ºC but you can cure as low as 100ºC. M-Bond
600 can also be used but it's not my favorite.
There are also the Buehler/Struers/Allied High Tech cold-setting
encapsulating epoxies. (I'm sure there are other vendors; these happen
to be the ones I use.) These epoxies are much harder than acrylics.
There only drawback for small sample sizes is their cure rate is
dependent on the mass of the epoxy; i.e., the less the epoxy mass, the
longer the cure time.

On 1/11/10 5:46 PM, jsilvey-at-tegal.com wrote:
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} Email: jsilvey-at-tegal.com
} Name: Judy Silvey
}
} Organization: Tegal Corp.
}
} Title-Subject: [Filtered] Epoxy
}
} Message: Epoxy fill material alternatives sought
}
}
}
} I am trying to characterize the deposition of polymer layers on the
} sidewalls of deep trenches in silicon. The polymer layers are soft
} and require polishing to capture the thickness and coverage of the
} deep trenches. I have been using acrylic-based epoxies to fill the
} trenches prior to polishing but these acrylic materials tend to
} deform during the polishing process and subsequent exposure to the
} beam of electrons in the SEM.
}
}
}
} I am looking for an epoxy or other fill material that will not deform
} when exposed to the polishing process or the SEM, and that does not
} require a high temperature anneal to cure. A room
} temperature-curable material is preferred if the cure time is less
} than ~24hrs. I suspect that a material that has a hardness similar
} to that of the surrounding silicon such a silicon dioxide or
} alumina-based epoxy is best but this has not been proven (in contrast
} to the acrylic epoxies that appear to be much softer.) I have
} considered using a spin-on-glass but the reported cure temperatures
} are too high, although a low temperature cure for an extended period
} might in theory work. (has anyone tried this?)
}
}
}
} Any ideas on good candidates for alternatives to the acrylic epoxies
} for filling deep trenches (~50um) in silicon that will tolerate the
} polishing process and the SEM exposure?
}
}
}
}
} Login Host: 72.245.193.146
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--
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Becky Holdford (r-holdford-at-ti.com)
972-995-2360
972-648-8743 (pager)
SC Packaging FA
Texas Instruments, Inc.
13536 N. Central Expressway MS 940
Dallas, TX 75243
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~


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From: colijn.1-at-osu.edu
Date: Mon, 11 Jan 2010 20:44:57 -0600
Subject: [Microscopy] Re: viaWWW: Epoxy fill material alternatives sought

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

Judy,

Can you mix either a fine metal or ceramic powder in with your epoxy to
help it hold up better? A metal filling may also help with charge
buildup. Sub-micron powder should be able to flow into the trenches.

Henk

jsilvey-at-tegal.com wrote:
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} Email: jsilvey-at-tegal.com
} Name: Judy Silvey
}
} Organization: Tegal Corp.
}
} Title-Subject: [Filtered] Epoxy
}
} Message: Epoxy fill material alternatives sought
}
}
}
} I am trying to characterize the deposition of polymer layers on the
} sidewalls of deep trenches in silicon. The polymer layers are soft
} and require polishing to capture the thickness and coverage of the
} deep trenches. I have been using acrylic-based epoxies to fill the
} trenches prior to polishing but these acrylic materials tend to
} deform during the polishing process and subsequent exposure to the
} beam of electrons in the SEM.
}
}
}
} I am looking for an epoxy or other fill material that will not deform
} when exposed to the polishing process or the SEM, and that does not
} require a high temperature anneal to cure. A room
} temperature-curable material is preferred if the cure time is less
} than ~24hrs. I suspect that a material that has a hardness similar
} to that of the surrounding silicon such a silicon dioxide or
} alumina-based epoxy is best but this has not been proven (in contrast
} to the acrylic epoxies that appear to be much softer.) I have
} considered using a spin-on-glass but the reported cure temperatures
} are too high, although a low temperature cure for an extended period
} might in theory work. (has anyone tried this?)
}
}
}
} Any ideas on good candidates for alternatives to the acrylic epoxies
} for filling deep trenches (~50um) in silicon that will tolerate the
} polishing process and the SEM exposure?
}
}
}
}
} Login Host: 72.245.193.146
} ---------------------------------------------------------------------------
}
} ==============================Original Headers==============================
} 18, 11 -- From zaluzec-at-microscopy.com Mon Jan 11 17:46:07 2010
} 18, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22])
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} 18, 11 -- From: jsilvey-at-tegal.com (by way of MicroscopyListserver)
} 18, 11 -- Subject: viaWWW: Epoxy fill material alternatives sought
} 18, 11 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed"
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}

--
Hendrik O. Colijn
www.ceof.ohio-state.edu
OSU Campus Electron Optics Facility colijn.1-at-osu.edu
040 Fontana Labs (614) 292-0674 (V)
116 W. 19th Ave. (614) 292-7523 (F)
Columbus, OH 43210

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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From: gary-at-gaugler.com
Date: Mon, 11 Jan 2010 22:59:35 -0600
Subject: [Microscopy] Photo copyrights

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

Form VA has been replaced by on-line CO for photographs.

Rules are the same, submission method has changed.

gary g.


==============================Original Headers==============================
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From: bengu-at-fen.bilkent.edu.tr
Date: Tue, 12 Jan 2010 01:47:07 -0600
Subject: [Microscopy] VP SEM for imaging biological samples

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


Dear All,

Couple of weeks ago I have sent an email regarding imaging of cancer cells
using SEM (thanx to Philip Oshel for his detailed procedure). I must admit
that I have now great respect for those of you going through such arduous
procedures for biological sample prep. I think I will cut corners and try
VP-SEM as there seems to be less sample prep involved.

I would like to find some general "how-to" and "dos and donts" on imaging
biological samples (human cells, tissue, etc) using VP mode (H2O or Dry)
on a CZ Evo40 SEM. I will appreciate if some who had experience using
VP-SEM can direct me the right way.

Best

Erman Bengu




=================================
Erman Bengu

Assistant Professor of Chemistry
Department of Chemistry
Bilkent University

Mailing Address:
Bilkent University,
Department of Chemistry,
06800, Bilkent, Ankara
Turkey

Office: SB #311
E-mail: bengu_AT_fen.bilkent.edu.tr
Phone (Office): +90 (312) 290-2153
(Lab1): +90 (312) 290-2663
(Lab2): +90 (312) 290-3332
Fax: +90 (312) 266-4068
Web: http://www.fen.bilkent.edu.tr/~bengu
==================================



=================================
Erman Bengu

Assistant Professor of Chemistry
Department of Chemistry
Bilkent University

Mailing Address:
Bilkent University,
Department of Chemistry,
06800, Bilkent, Ankara
Turkey

Office: SB #311
E-mail: bengu_AT_fen.bilkent.edu.tr
Phone (Office): +90 (312) 290-2153
(Lab1): +90 (312) 290-2663
(Lab2): +90 (312) 290-3332
Fax: +90 (312) 266-4068
Web: http://www.fen.bilkent.edu.tr/~bengu
==================================





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23, 22 -- Subject: VP SEM for imaging biological samples
23, 22 -- From: "Erman Bengu" {bengu-at-fen.bilkent.edu.tr}
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From: David.Patton-at-uwe.ac.uk
Date: Tue, 12 Jan 2010 06:25:35 -0600
Subject: [Microscopy] VP SEM for imaging biological samples

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

Standard preparation of the cells including metal coating will give you images that are easier to interpret and at higher magnifications, if required.

Wet cells are difficult to image beyond very low magnification unless you have a field emission ESEM or VP SEM. The surrounding water film has to be carefully removed without dehydrating the cells. They are beam sensitive and easily damaged.

A good paper on wet cell imaging is:

Kirk SE, Skepper JN Donald AM. 2009 Application of environmental scanning electron microscopy to determine surface structure. J Microsc 233:205-244.


Dave

-----Original Message-----
X-from: bengu-at-fen.bilkent.edu.tr [mailto:bengu-at-fen.bilkent.edu.tr]
Sent: 12 January 2010 07:51
To: David Patton


Dear All,

Couple of weeks ago I have sent an email regarding imaging of cancer cells
using SEM (thanx to Philip Oshel for his detailed procedure). I must admit
that I have now great respect for those of you going through such arduous
procedures for biological sample prep. I think I will cut corners and try
VP-SEM as there seems to be less sample prep involved.

I would like to find some general "how-to" and "dos and donts" on imaging
biological samples (human cells, tissue, etc) using VP mode (H2O or Dry)
on a CZ Evo40 SEM. I will appreciate if some who had experience using
VP-SEM can direct me the right way.

Best

Erman Bengu




=================================
Erman Bengu

Assistant Professor of Chemistry
Department of Chemistry
Bilkent University

Mailing Address:
Bilkent University,
Department of Chemistry,
06800, Bilkent, Ankara
Turkey

Office: SB #311
E-mail: bengu_AT_fen.bilkent.edu.tr
Phone (Office): +90 (312) 290-2153
(Lab1): +90 (312) 290-2663
(Lab2): +90 (312) 290-3332
Fax: +90 (312) 266-4068
Web: http://www.fen.bilkent.edu.tr/~bengu
==================================



=================================
Erman Bengu

Assistant Professor of Chemistry
Department of Chemistry
Bilkent University

Mailing Address:
Bilkent University,
Department of Chemistry,
06800, Bilkent, Ankara
Turkey

Office: SB #311
E-mail: bengu_AT_fen.bilkent.edu.tr
Phone (Office): +90 (312) 290-2153
(Lab1): +90 (312) 290-2663
(Lab2): +90 (312) 290-3332
Fax: +90 (312) 266-4068
Web: http://www.fen.bilkent.edu.tr/~bengu
==================================





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From: Nicola.Weston-at-nottingham.ac.uk
Date: Tue, 12 Jan 2010 06:57:43 -0600
Subject: [Microscopy] RE: VP SEM for imaging biological samples

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

Other interesting papers:

Stokes DJ, Rea SM, Best SM, Bonfield W. 2003. Electron microscopy of
mammalian
cells in the absence of fixing, freezing, dehydration, or specimen
coating. Scanning 25:181-184.

McKinlay K, Allison FJ, Scotchford C, Grant DM, Oliver JM, King JR, Wood
JV,
Brown PD. 2004. Comparison of environmental scanning electron microscopy
with high vacuum scanning electronmicroscopy as applied to the
assessment of
cell morphology. J Biomed Mater Res A 69(2):359-366.

I have imaged a fair few cell cultures in a FEG ESEM but always on fixed
cells. Buffer salts can cause problems so samples need to be thoroughly
washed.
It is possible to use lower voltages with the FEG, allowing more cell
surface detail to be seen and reducing beam damage. Sample needs to be
cooled and kept wet during pumpdown so I always put excess water
droplets around the sample. The film of water covering cells needs to be
carefully removed and you do not always need to be at 100% RH due to the
salts within the cells, but dehydration will occur rapidly if pressure
is taken too low. It's a fine balance and you have limited observation
time before you are looking at dehydrated cells, however you may well
get images of features damaged by conventional prep techniques.

Best of luck
Nicola
This message has been checked for viruses but the contents of an attachment
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From: Rob.Bowen-at-caddock.com
Date: Tue, 12 Jan 2010 09:17:05 -0600
Subject: [Microscopy] Re: viaWWW: Epoxy fill material alternatives sought

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

I'm not quite clear on your sample. It sounds like you have trenches in silicon that are covered with a polymer that you want to then cover/fill with epoxy. Is that right?

I agree with the others that a two-part epoxy is probably more robust than the acrylics. Without knowing your polishing process, I think you would find many epoxies will stand up to polishing. Edge retention will be an issue, but the epoxy will probably be harder than your polymer.

I don't know how well fillers would work with the epoxy. I would be a bit afraid that the filler would increase the viscosity and prevent the trench from being well filled. On the other hand, the filler could provide contrast so that you can differentiate the epoxy from the polymer.

Contrast between the epoxy and polymer will be an issue. The different hardness should give polishing relief (sort of an etching) so that you can distinguish the two phases. Otherwise, the polymer and epoxy may appear to be one, continuous phase. If your polymer does not have an additional element, you may need to dope the epoxy so that you have some contrast between the phases.

Warren S.

-----Original Message-----
X-from: colijn.1-at-osu.edu [mailto:colijn.1-at-osu.edu]
Sent: Monday, January 11, 2010 8:45 PM
To: wesaia-at-iastate.edu


--
Judy,
A UV cure material might be worth considering. Dymax and I sort of
recall Lord Chemical have those, among others. UV cure is even used for wood
floor finishes, so there ought to be something hard enough.

Rob Bowen

Robert C. Bowen
Research Scientist
Caddock Electronics, Inc
rob.bowen-at-caddock.com
http://www.caddock.com


} From: {jsilvey-at-tegal.com}
} Reply-To: {jsilvey-at-tegal.com}
} Date: Mon, 11 Jan 2010 17:48:52 -0600
} To: {rob.bowen-at-caddock.com}
} Subject: [Microscopy] viaWWW: Epoxy fill material alternatives sought
}
}
}
}
} ----------------------------------------------------------------------------
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} Email: jsilvey-at-tegal.com
} Name: Judy Silvey
}
} Organization: Tegal Corp.
}
} Title-Subject: [Filtered] Epoxy
}
} Message: Epoxy fill material alternatives sought
}
}
}
} I am trying to characterize the deposition of polymer layers on the
} sidewalls of deep trenches in silicon. The polymer layers are soft
} and require polishing to capture the thickness and coverage of the
} deep trenches. I have been using acrylic-based epoxies to fill the
} trenches prior to polishing but these acrylic materials tend to
} deform during the polishing process and subsequent exposure to the
} beam of electrons in the SEM.
}
}
}
} I am looking for an epoxy or other fill material that will not deform
} when exposed to the polishing process or the SEM, and that does not
} require a high temperature anneal to cure. A room
} temperature-curable material is preferred if the cure time is less
} than ~24hrs. I suspect that a material that has a hardness similar
} to that of the surrounding silicon such a silicon dioxide or
} alumina-based epoxy is best but this has not been proven (in contrast
} to the acrylic epoxies that appear to be much softer.) I have
} considered using a spin-on-glass but the reported cure temperatures
} are too high, although a low temperature cure for an extended period
} might in theory work. (has anyone tried this?)
}
}
}
} Any ideas on good candidates for alternatives to the acrylic epoxies
} for filling deep trenches (~50um) in silicon that will tolerate the
} polishing process and the SEM exposure?
}
}
}
}
} Login Host: 72.245.193.146
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From: bigelow-at-umich.edu
Date: Tue, 12 Jan 2010 11:49:04 -0600
Subject: [Microscopy] [Microscopy}RE; Hard epoxies

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Judy:

The PC-11 and PC-7 epoxies that are available in most hardware stores
in the U.S. (I don't know where you are located), are both hard
enough to drill, file and machine when fully hardened (less than one
day). They are also very strong and therefore very useful around the
lab. Also, Torr Seal ultra high vacuum epoxy from Gatan (and
similar products available from EM supply companies such as SPI) are
also quite hard when cured. All cure at room temperature.

--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From: DusevichV-at-umkc.edu
Date: Tue, 12 Jan 2010 13:39:32 -0600
Subject: [Microscopy] RE: VP SEM for imaging biological samples

Contents Retrieved from Microscopy Listserver Archives
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Using dry VP mode for wet cells is useless. The same results (with
better quality of pictures) you can get by washing cells, air drying
them and coating with sputter coater. But you'll have a lot of
artifacts.

You have to fix cells. Even for "wet VP" mode it is better to fix, and
in most cases it is the only possible way to observe wet cells. Anyway,
wet cells observation is the very tough task. Much (much!) easier is to
go with standard procedure, recommended by Philip Oshel.

You can try to simplify the protocol a little:
Fix with 2% gluteraldehyde in buffer. Twice wash with water. Dehydrate
for 15 min in each of ethanol solutions: 33%, 67%, 95% and twice in
100%. If you have access to Critical point drier, use it. Otherwise
replace 100% ethanol with HMDS, and after 10 min pipette HMDS out,
leaving just enough to cover your specimen. Let it air dry in the hood.
Coat with sputter coater.

May be you can even skip HMDS, and just air dry specimen after ethanol.
You will have some artifacts, like cracks in cells and additional
shrinkage, but results could be acceptable.

Good luck,

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

} -----Original Message-----
} From: bengu-at-fen.bilkent.edu.tr [mailto:bengu-at-fen.bilkent.edu.tr]
} Sent: Tuesday, January 12, 2010 1:48 AM
} To: Dusevich, Vladimir
} Subject: [Microscopy] VP SEM for imaging biological samples
}
}
}
}
}
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}
}
} Dear All,
}
} Couple of weeks ago I have sent an email regarding imaging of cancer
} cells
} using SEM (thanx to Philip Oshel for his detailed procedure). I must
} admit
} that I have now great respect for those of you going through such
} arduous
} procedures for biological sample prep. I think I will cut corners and
} try
} VP-SEM as there seems to be less sample prep involved.
}
} I would like to find some general "how-to" and "dos and donts" on
} imaging
} biological samples (human cells, tissue, etc) using VP mode (H2O or
} Dry)
} on a CZ Evo40 SEM. I will appreciate if some who had experience using
} VP-SEM can direct me the right way.
}
} Best
}
} Erman Bengu
}
}
}
}
} =================================
} Erman Bengu
}
} Assistant Professor of Chemistry
} Department of Chemistry
} Bilkent University
}
} Mailing Address:
} Bilkent University,
} Department of Chemistry,
} 06800, Bilkent, Ankara
} Turkey
}
} Office: SB #311
} E-mail: bengu_AT_fen.bilkent.edu.tr
} Phone (Office): +90 (312) 290-2153
} (Lab1): +90 (312) 290-2663
} (Lab2): +90 (312) 290-3332
} Fax: +90 (312) 266-4068
} Web: http://www.fen.bilkent.edu.tr/~bengu
} ==================================
}
}
}
} =================================
} Erman Bengu
}
} Assistant Professor of Chemistry
} Department of Chemistry
} Bilkent University
}
} Mailing Address:
} Bilkent University,
} Department of Chemistry,
} 06800, Bilkent, Ankara
} Turkey
}
} Office: SB #311
} E-mail: bengu_AT_fen.bilkent.edu.tr
} Phone (Office): +90 (312) 290-2153
} (Lab1): +90 (312) 290-2663
} (Lab2): +90 (312) 290-3332
} Fax: +90 (312) 266-4068
} Web: http://www.fen.bilkent.edu.tr/~bengu
} ==================================
}
}
}
}
}
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From: mckee-at-helix.mgh.harvard.edu
Date: Tue, 12 Jan 2010 19:16:07 -0600
Subject: [Microscopy] viaWWW: uranyl acetate crystals

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Email: mckee-at-helix.mgh.harvard.edu
Name: Mary McKee

Organization: MGH

Title-Subject: [Filtered] uranyl acetate crystals

Message: Dear List,

I am having a problem with uranyl acetate crystals on tissue culture
cells that I stain, enbloc after OsO4. I thoroughly rinse the cells
after OsO4, first in sodium cacodylate buffer and then in DH2O before
the Ua step. After Ua, I rinse in DH2O, but I seem to get the
crystals on cells (not on tissue). In cells with only OsO4, there
are no crystals. Does anyone have any ideas? I filter everything.
Thanks.

Mary

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From: a.chuvilin-at-microscopist.ru
Date: Wed, 13 Jan 2010 03:39:17 -0600
Subject: [Microscopy] TEM technician position opening

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


Judy:

As others say, we have had very good luck with 2 part epoxies in our ground cross sections. We use the Buehler "Epoxicure." Edge retention with Si is good. And, there is more in the fillers area. Recently I added Carbon-Graphite filler at 4 weight percent to the mixed up Epoxicure. This accomplishes two things, it makes great contrast for optical light microscopy and it does a beautiful job mitigating charge in the SEM! I use a fine carbon powder, the tailings from ground-up Ted Pella graphite rods. (The tailings occur when we sharpen the tips of these graphite rods for our Cressington carbon coater).

Good luck.

Pete Eschbach
Nanoindenter/X section/Electron Microscope Engineer
Hewlett Packard Corp.


-----Original Message-----
X-from: wesaia-at-iastate.edu [mailto:wesaia-at-iastate.edu]
Sent: Tuesday, January 12, 2010 7:25 AM
To: Eschbach, Peter

I'm not quite clear on your sample. It sounds like you have trenches in silicon that are covered with a polymer that you want to then cover/fill with epoxy. Is that right?

I agree with the others that a two-part epoxy is probably more robust than the acrylics. Without knowing your polishing process, I think you would find many epoxies will stand up to polishing. Edge retention will be an issue, but the epoxy will probably be harder than your polymer.

I don't know how well fillers would work with the epoxy. I would be a bit afraid that the filler would increase the viscosity and prevent the trench from being well filled. On the other hand, the filler could provide contrast so that you can differentiate the epoxy from the polymer.

Contrast between the epoxy and polymer will be an issue. The different hardness should give polishing relief (sort of an etching) so that you can distinguish the two phases. Otherwise, the polymer and epoxy may appear to be one, continuous phase. If your polymer does not have an additional element, you may need to dope the epoxy so that you have some contrast between the phases.

Warren S.

-----Original Message-----
X-from: colijn.1-at-osu.edu [mailto:colijn.1-at-osu.edu]
Sent: Monday, January 11, 2010 8:45 PM
To: wesaia-at-iastate.edu

PLEASE DO NOT REPLY TO THIS EMAIL; SEE CONTACT INFORMATION BELOW.

CIC nanoGUNE Consolider, located in San Sebastian, Basque Country
(Spain), is a R&D center created recently with the mission of
conducting basic and applied world-class research in nanoscience and
nanotechnology, fostering training and education excellence, and
supporting the growth of a nanotechnology-based industry.

An electron microscopy facility is currently being established to
accompany nanoGUNE´s already ongoing research activities by means of a
high-level structural characterization laboratory, including Cs
corrected TEM, FIB and ESEM. In order to provide qualified assistance
to the Staff Microscopist that is managing these facilities, we have
an immediate opening for an Electron Microscopy Technician. We are
searching for a highly motivated and creative person with a solid
technical/engineering background, mechanical and electronic design and
construction skills, and experience in operating and maintaining TEM
sample preparation facilities.


Requirements for this position are:

- suitable technical or engineering education

- 5 years hands-on experience in TEM sample preparation techniques

- basic experience in TEM, SEM, FIB operation

- good English communication skills


Responsibilities will include:

- general instrument maintainance

- establishing and maintaining a sample preparation lab

- hands-on samples preparation

- equipment and consumables purchase

- engineering assistance of ongoing research and projects



We will offer a competitive salary commensurable to educational
qualifications and working experience of the candidate. This position
is envisioned to be a permanent position after an appropriate
evaluation period.

Interested individuals should send a letter of application and their
curriculum vitae under Ref: "TEM technician application" to nano-at-nanogune.eu

Closing date: 01 March 2010.



--------------------------------
Andrey Chuvilin
Ikerbasque Research Professor
TEM Staff Scientist
a.chuvilin-at-nanogune.eu

CIC nanoGUNE Consolider
Tolosa Hiribidea, 76
E-20018 Donostia - San Sebastian
Spain
+34 943 574 023
http://www.nanogune.eu
--------------------------------



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From: gary.nichols-at-pfizer.com
Date: Wed, 13 Jan 2010 08:38:31 -0600
Subject: [Microscopy] viaWWW: Measuring specimen current in VP mode

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Email: gary.nichols-at-pfizer.com
Name: Gary Nichols

Organization: Pfizer Global R&D (UK)

Title-Subject: [Filtered] Measuring specimen current in VP mode

Question: Dear Listers,

Do any of you have some guidance on how to measure the specimen/probe
current using a Faraday cup whilst in VP mode? I am using a Zeiss
SUPRA 40VP FE-SEM and have found that the measured current depends
upon the gas pressure in the chamber and on the bias voltage applied
to the VPSE detector.

Many thanks in advance.
Gary

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From: rgoddard-at-valdosta.edu
Date: Wed, 13 Jan 2010 12:23:49 -0600
Subject: [Microscopy] Blind Student in Biology Lab

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I don’t often send messages to the list but enjoy reading the discussions. 
I am at a loss as to how to handle a current problem we have in our
non-majors biology teaching lab.  A colleague came to me yesterday with the
problem of having a totally blind student (since birth) taking our
non-majors biology lab.  This is a very visual lab and we use student light
microscopes for many of the exercises that we use.  Can anyone make
suggestions on how we might customize a lab experience for this student to
make her experience rewarding in this class?  We’ve talked about
clay-modeling for tactile models of cells but I’ve never had to solve a
problem like this.

Thanks in advance!  I’ll be glad to post any responses I get to the list in
summary form in a few days.

Russ Goddard
Valdosta State University
Biology Dept.



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From: TrogadisJ-at-smh.toronto.on.ca
Date: Wed, 13 Jan 2010 12:38:16 -0600
Subject: [Microscopy] we just need a couple of fluorescent images

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

Dear Owners of Imaging Equipment :

Please excuse the non-conventional nature of this post.

One of the PI's coming to our Imaging Facility would like some preliminary data from 5-6 slides and we don't have the right combination of hardware to do this. If it works out, we will invest in upgrades, in the meantime, I wondered whether any of you could help. We could send you the slides - and pay, of course.

She is looking at ICG dye (indocyanine green), an IR dye used commonly in ophthalmology to study circulation and equipment is available to routinely image this dye in intact eyes. In this study, following ICG labelling and image capture, the eyes were sectioned, mouse, I think, and now they want to look at the same specimen at the microscopic level. We don't have the equipment to do this.

ICG excitation : 805, emission : 835.
A xenon light source (which we do have) would excite the dye, however we don't have proper filters. A monochromatic CCD camera usually has response in longer wavelengths (colour cameras have IR filters), however the efficiency of ours at } 800nm is very low.

Can anyone help?
Thank you in advance.
Judy

Judy Trogadis
Bio-Imaging Coordinator
St. Michael's Hospital, 7Queen
30 Bond St.
Toronto, ON M5B 1W8, Canada
ph: 416-864-6060 x6337
pager: 416-685-9219
fax: 416-864-5046
trogadisj-at-smh.toronto.on.ca




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From: oshel1pe-at-cmich.edu
Date: Wed, 13 Jan 2010 12:44:31 -0600
Subject: [Microscopy] Re: Blind Student in Biology Lab

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

Russ,

I haven't had a blind student, but have a
thought: the model idea is good, especially since
there are commercial 3D models. The student could
then be given a ball of plasticine and asked to
form models of cells, organelles, different leaf
types, and so on after examing the model, or the
real object (real leaf, etc.). She could then be
asked to modify the model she made to show how a
different e.g. leaf is shaped differently. Such a
transformation helps get across the idea of
morphology adaptations to environmental
differences. (Drip-tips on leaves, and so on.)

Another idea is to lay a sheet of aluminum foil**
on a smooth, fresh sheet of wax, and use a blunt
dissecting probe (or other burnisher) to "draw"
forms on the foil, producing indentations (ridges
when flipped over) that could be "read" as
Braille-like pictures.
Another student could draw a picture of the
subject while the blind student rested her hand
on the 1st student's drawing hand, and would then
work to duplicate the image with the foil.

**Or maybe something else, tougher than Al foil, but equally malleable.

I'll put on my other hat here, and if anyone does
have experience with teaching such a visual
subject to blind students, it would make an
excellent article for Microscopy Today.

Phil

} I donít often send messages to the list but enjoy reading the discussions.Ý
} I am at a loss as to how to handle a current problem we have in our
} non-majors biology teaching lab.Ý A colleague came to me yesterday with the
} problem of having a totally blind student (since birth) taking our
} non-majors biology lab.Ý This is a very visual lab and we use student light
} microscopes for many of the exercises that we use.Ý Can anyone make
} suggestions on how we might customize a lab experience for this student to
} make her experience rewarding in this class?Ý Weíve talked about
} clay-modeling for tactile models of cells but Iíve never had to solve a
} problem like this.
}
} Thanks in advance!Ý Iíll be glad to post any responses I get to the list in
} summary form in a few days.
}
} Russ Goddard
} Valdosta State University
} Biology Dept.
--
Philip Oshel
Technical Editor, Microscopy Today
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576


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From: bozzola-at-siu.edu
Date: Wed, 13 Jan 2010 12:47:35 -0600
Subject: [Microscopy] Re: Blind Student in Biology Lab

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

It is an admirable thing that you are attempting to accomplish and I
salute you.

Are there any models available of cells that are similar to the
anatomical, plaster models of humans and plants? They should consist
of layers that come apart.

In the past, for a lower level course, I made a cell out of jello with
the individual components (nuclei, nucleoli, mitochondria, plastids,
etc) composed of various fruits: peach = nucleus, peach pit =
nucleolus, banana = mitochondrion, grapes = vesicles, etc. It's great
because you could discuss sol/gel phenomena, migration of organelles,
including chromosomes (licorice strands).

The day after the hands on experience, I would bring in a clean,
gelled cell and we would eat the cell, usually with whipped cream
(aka, extracellular matrix). More mitochondria.... please.

Obviously, it's lunch time here.......


--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730



On Wed, Jan 13, 2010 at 12:24 PM, {rgoddard-at-valdosta.edu} wrote:
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} I don’t often send messages to the list but enjoy reading the discussions.
} I am at a loss as to how to handle a current problem we have in our
} non-majors biology teaching lab.  A colleague came to me yesterday with the
} problem of having a totally blind student (since birth) taking our
} non-majors biology lab.  This is a very visual lab and we use student light
} microscopes for many of the exercises that we use.  Can anyone make
} suggestions on how we might customize a lab experience for this student to
} make her experience rewarding in this class?  We’ve talked about
} clay-modeling for tactile models of cells but I’ve never had to solve a
} problem like this.
}
} Thanks in advance!  I’ll be glad to post any responses I get to the list in
} summary form in a few days.
}
} Russ Goddard
} Valdosta State University
} Biology Dept.


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From: connellyps-at-nhlbi.nih.gov
Date: Wed, 13 Jan 2010 13:28:19 -0500
Subject: [Microscopy] Blind Student in Biology Lab

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

Russ,

I was at Temple University Medical Center in the early 70's. At this time
there was a totally blind medical student who had to take all the courses
and practicals that anyone else had to participate in. I was told that he
had to memorize all the details of histology slides so that when someone
described a slide to him he could tell the tissue type etc. He went on to be
a Psychiatrist I believe.

Being the daughter of a totally blind father (WWII Vet.) I know that there
is an amazing amount of things that blind people can do - sometimes in a
slightly different manner than what a sighted person may expect.

Since you are in a University this student has certainly gone through high
school science classes already. I would strongly suggest that you speak
with her and find out what had worked for her in previous science classes.
There is a lot more to lab than looking at things. She can hold, pour,
assist, etc. I bet she'll be great at recording the results that her lab
partner could be discussing with her. She may be excellent in drawing
conclusions.
See where I am going with this?

I have seen students that could not look at a dissection, pass that lab
because of the cooperation of a lab partner who did not mind the actual
procedure. It happens.

I'm sure you will both benefit from the class!
Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E ­ Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-480-6560
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and can be freely shared
and reproduced.




X-from: {rgoddard-at-valdosta.edu}
Reply-To: {rgoddard-at-valdosta.edu}

I don¹t often send messages to the list but enjoy reading the discussions. 
I am at a loss as to how to handle a current problem we have in our
non-majors biology teaching lab.  A colleague came to me yesterday with the
problem of having a totally blind student (since birth) taking our
non-majors biology lab.  This is a very visual lab and we use student light
microscopes for many of the exercises that we use.  Can anyone make
suggestions on how we might customize a lab experience for this student to
make her experience rewarding in this class?  We¹ve talked about
clay-modeling for tactile models of cells but I¹ve never had to solve a
problem like this.

Thanks in advance!  I¹ll be glad to post any responses I get to the list in
summary form in a few days.

Russ Goddard
Valdosta State University
Biology Dept.

==============================End of - Headers==============================




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From: joelsheffield-at-gmail.com
Date: Wed, 13 Jan 2010 14:18:25 -0600
Subject: [Microscopy] re: Blind Student in Lab

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

I distributed this query to a Biology Teaching list that I monitor.  Here is a set of replies that arrived
within an hour of my posting:


X-from Nancy Ruggeri {nruggeri-at-wisc.edu}
This sounds like a fabulous opportunity to engage your students and 
create community in your classroom. If you have them working in 
groups, you could have this student's lab partners describe what they 
see in the microscopes, and develop ways to help each other 
communicate the visual aspects of the lab. Interesting elements could 
come out about form and function... I think it sounds like a great 
"teachable moment" that can benefit everyone involved.  Good luck!


Nancy


X-from:   "Morris, Amy" { amorris-at-hastings.edu }
The tactile modeling sounds great.


Here is my experience. During my master's program I made extra money by
working in the student services area of the university. One of my jobs one
semester was to attend lab with a blind student. I did all the "looking at
things" through the microscope. My responsibility to the student was to
describe what I saw. Maybe some combo of using a student helper to
describe what is being observed under the microscope paired with
activities that use other senses.


Amy Morris, Ph.D.
Associate Professor of Biology
Hastings College
710 N. Turner Ave.
Hastings, NE 68901
402-461-7745


X-from:   John La Duke { john.laduke-at-und.edu }
Our Student Support Services began creating braille handouts 
including raised surface images for us some yrs ago.  The student was  in
the lab for a few sessions but never finished.  The materials are
difficult to create and close interaction with the professional was
necessary.  If it would help, I can look to see if I still have any  of
it.


John
X-from:   "Stacey Kiser" { kisers-at-lanecc.edu }
I have never had a blind-since-birth student, but I had two severely visually impaired students in
the non-majors cell bio classes. In one case, I had the microscope hooked up to a small t.v. set so
the student could use the microscope by herself for the first time in her bio experience. She loved
it. We don't seem to clean out our stockroom very often, so we have lots of old models that are in
3D - animal and plant cells, mitosis, etc. The students were able to get a lot from the model by
feeling it while a lab partner described/named structures for them. We experimented with clay,
string, and puffy paint for graphs (some success). Our Disability Resources office now has a
printer that can take an image and turn it into a 3D print-out on special paper. Both students had
their own laptops with software that allowed them to view web sites. It helped me justify why we
needed a wireless hub in our end of the building.




My best success came from getting the students in a good lab group. In both cases, the students
around the visually impaired students learned a lot by helping out. In one case they would had the
pipettes over so she could feel them, and included her in the lab to the fullest extent.




Stacey




Stacey Kiser
Biology Instructor
Science Division
Building 16, Office 162H
Lane Community College
Eugene, OR 97405


I'll send more if I get them.












On 13 Jan 2010 at 12:31, rgoddard-at-valdosta.edu wrote:


}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} I don´t often send messages to the list but enjoy reading the discussions. 
} I am at a loss as to how to handle a current problem we have in our
} non-majors biology teaching lab.  A colleague came to me yesterday with the
} problem of having a totally blind student (since birth) taking our
} non-majors biology lab.  This is a very visual lab and we use student light
} microscopes for many of the exercises that we use.  Can anyone make
} suggestions on how we might customize a lab experience for this student to
} make her experience rewarding in this class?  We´ve talked about
} clay-modeling for tactile models of cells but I´ve never had to solve a
} problem like this.
}
} Thanks in advance!  I´ll be glad to post any responses I get to the list in
} summary form in a few days.
}
} Russ Goddard
} Valdosta State University
} Biology Dept.
}

--
Joel B. Sheffield, Ph.D.
Biology Department, Temple University
1900 North 12th Street
Philadelphia, PA 19122
jbs-at-temple.edu
(215) 204 8839, fax (215) 204 0486
http://astro.temple.edu/~jbs



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From: joelsheffield-at-gmail.com
Date: Wed, 13 Jan 2010 14:42:27 -0600
Subject: [Microscopy] Re: Blind Student in Lab

Contents Retrieved from Microscopy Listserver Archives
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Hi All,

Some more interesting replies.:
Joel

X-from: "Bowen, Jeffery" {jabowen-at-bridgew.edu}

I, too, had this problem with a student. Fortunately, we had some 3D
models of cells that the student could manipulate. Additionally, we also
found that the "raised paint" that kids use to draw on their t-shirts and
things was also very effective (one of the department moms said it is
called liquid embroidery paint). We would draw the cells (or stage of
mitosis, etc) and the student could 'feel' what was going on. Also, BSC
has a history for training educators (we were the first Normal College in
the US), I was lucky to find a good and dedicated student who was in the
Education department and wanted the experience of working with a special
needs student. It worked out very well. The student would use that paint
stuff to draw what she saw in the microscope...things like amoeba or
paramecium, the usual stuff. And, she would describe what she saw to the
student. Both benefited from this interaction. I also benefited from
having this young man in my lecture and lab as I found myself trying to be
more descriptive during my lectures and not solely relying on the
diagrams. I'd be happy to elaborate more if you need!

Jeff
P.S. I also had the same student in a non-major Human Sexuality class. He
and I had a running joke that he couldn't use the class to approach women
with the line that he had to study anatomy for this class and needed a
model.

X-from: "Tonna Harris-Haller" {tonna-at-mail.bio.tamu.edu}

Do you know if the university has a disabilities services program? We had
two blind students in our majors level first semester course last
semester, and will have one in our second semester course this term. We
also have been struggling with a way to teach blind students in a
microscope intensive course.

We are considerably aided by our University's Disabilities Services
Program. So far this is what we've come up with.

1. Label the microscope (and any other equipment) with braille
characters and require the student to recognize the structure and
function of the key elements of the microscope. We bought a braille
hand labeler, and have complete lab set with braille labels (including
glassware so the student can setup some experiments). It's time
consuming, but we now have the labeled materials, and I have a staff
member who can read braille as a result of all her labeling.

2. Encourage the lab group that includes the blind student to engage in
descriptive interaction. Get them to help teach the student what the
results of an experiment look like, or what a slide looks like etc. It
helps if the lab instructor has an assistant. The instructor can take
care of the rest of the class, while the assistant circulates to help and
keeps an eye out to assist the blind student as necessary.

3. Meet with the blind student before class to physically show them
around the lab so they know where everything is located.

4. Create high contrast (simplified) diagrams that contain key
elements. These drawings can be rendered into a three dimensional image
via a thermal machine and provided to the student to study in lieu of
looking at microscope slides. Our disabilities services department has a
machine that can take black and white drawings and render them into
three-dimensional images. If your university doesn't have one, you may
have to select the key images and draw them yourself (see suggestion #5).
We plan to take use this strategy to get through the slide work for our
courses.

5. Where a thermal paper machine is not available, or it is NOT
possible to render a diagram into a 3-dimensional image, use some
commonly accessible art supplies such as raised tape, three dimensional
dots, and puff paint to create tactile images. We managed to teach a
forensic lab complete with fingerprints and DNA electrophoresis results to
our blind students last semester using these techniques.

6. Rely on touch, taste, sound, and smell as much as possible. Try to
think of ways to teach the topic that are not exclusively visual. Use
three-dimensional models, or actual specimens that are safe to touch
instead of slides where possible. Label the models with braille
characters. If the actual word is too long to use, tag the model with
braille characters such as A, B, C ... and then make a braille key to go
with the model.

7. Give quizzes and exams separately from the rest of the class. The
blind student will require more time. If you can convert the tests to
braille, that is great, but if not someone will need to read the questions
and record the answers. Even if the question is in braille there may
still be a need to record the answer. Also if a lab practical is
involved, someone will need to escort the student through the stations,
describe the setups, and guide the student to the tactile portions of the
setup.

8. Give the student the text and lab manual enough time in advance to
convert them to auditory or braille documents. Our disabilities services
program does this for us, but some students have individual text readers.

Here are a few other resources you might try:

http://www.washington.edu/doit/Faculty/Strategies/Academic/Science/

http://www.tsbvi.edu/math/tools-blind.htm

http://www.aph.org/

http://www.afb.org/Section.asp?SectionID=44&TopicID=16&SubTopicID=35&Docum
entID=476

Good luck.

Tonna




Tonna Harris-Haller
Associate Director - Lower Division Biology Program
Biology Dept.
Texas A&M University
College Station, Tx. 77843-3258

phone: 979-845-4606
fax; 979-458-2030



--
Joel B. Sheffield, Ph.D.
Biology Department, Temple University
1900 North 12th Street
Philadelphia, PA 19122
jbs-at-temple.edu
(215) 204 8839, fax (215) 204 0486
http://astro.temple.edu/~jbs


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From: awelford-at-salud.unm.edu
Date: Wed, 13 Jan 2010 14:53:10 -0600
Subject: [Microscopy] viaWWW: processing dynamic cell culture for TEM

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Email: awelford-at-salud.unm.edu
Name: Angela Welford

Organization: University of New Mexico

Title-Subject: [Filtered] processing dynamic cell culture for TEM

Message: I have been asked to prepare a two-cell system for TEM to
observe a temporary and not very strong attachment between the two
cell types in culture. One of the cell lines adheres to the
substrate while the other floats and makes temporary attachments with
the adherent cells (as observed by time lapse video). The challenge
is to collect the cells in a manner that preserves at least some of
the attachments. Has anyone any experience with such a system, or
any suggestions?

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From: schooley-at-mcn.org
Date: Wed, 13 Jan 2010 15:30:37 -0600
Subject: [Microscopy] Re: Blind Student in Biology Lab

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

You've received a lot of clever, sensitive & helpful replies to your
inquiry. I'd like to suggest that you buy a book to share with your
lab staff (you can find it online for little more than the postage).
Geeraty Vermeij is arguably the world's leading authority on
molluscan morphological development, and he's been blind since early
childhood. He's on the U. of California at Davis faculty, and
"Priviledged Hands: a Scientific Life" is his autobiography.
--
Caroline Schooley
Project MICRO Coordinator
Microscopy Society of America
45301 Caspar Point Road, Box 117
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.microscopy.org/ProjectMICRO

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From: protrain-at-emcourses.com
Date: Thu, 14 Jan 2010 08:32:56 -0600
Subject: [Microscopy] State of the Art SEM

Contents Retrieved from Microscopy Listserver Archives
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Hi Listers

Pre Christmas I spent several months south of the Equator working with some
very good scientists on state of the art SEM. All of these instruments were
FEG, with both in and out of lens secondary electron detectors. Having
spent the last week writing up reports it suddenly struck me that the
techniques I introduced should have been part of the standard training for
this type of instrument.

I was one of the first people in the world to run a double detector
instrument when I worked on a pre production SS Series instrument at ISI.
My task was to look at all the options made available with a double detector
instrument, the advantages and disadvantages of the different detectors with
different types of specimen. With that information, as I gradually
developed an understanding of the generation of the signals I was receiving,
I was able to acquire exactly the signal mix which optimised the image for
the task in hand.

Over these past months it was very clear that the people I worked with just
knew that using the upper and short working distances gave them high
resolution, the subtleties of double detection instruments were unknown
because they had not been taught how to use the instrument correctly. I
think of the state of the art SEM as a racing car; my other life. There are
times, when it is easy to do so, that you go flat out because you have so
much potential, but there are other times when flat out makes the task much
more difficult. That is state of the art SEM, you have so much resolution
available but you really need other instrument attributes to obtain the
perfect image. So often I witnessed a quest for resolution when, with the
potential of the instrument, the chronic charging of the specimen should
have taken priority and I don't just mean turning down the accelerating
voltage.

So the point of this note, do people suffer from the same problems in the
northern hemisphere in that the subtleties of operation are ignored when
manufacturers train operators on state of the art FEG SEM? There are more
FEG SEM in the northern hemisphere so does that mean the knowledge of the
manufacturer's staff is of a higher level?

Thoughts?

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



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From: freym2-at-rpi.edu
Date: Thu, 14 Jan 2010 10:53:44 -0600
Subject: [Microscopy] State of the Art SEM

Contents Retrieved from Microscopy Listserver Archives
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Hi Steve,

It is a very interesting point you bring up. I spent nearly 10 years working
for one the manufactures of High res- FESEM's as an applications specialist.
The topics/concepts that were emphasized in training were often driven by
the demands of the customer. This seemed to be more true in an onsite
training rather than those we would provide to a mixed group (people from
different companies) at our offices/demo labs. This maybe right or wrong
depending on where you stand on the following statements, "The customer
doesn't know either what he really wants, or what he really needs." And the
"Customer is always right." I think that it is often the case that the
applications people are being driven by trying to meet the customers
demands/expectations, "I purchased a HI-RES FESEM, I demand HI-RES 100% of
the time." Many of the FESEM's that we sold were into an "industrial"
environment where good and knowledgeable people were always under the gun,
and had an end result they needed to meet. Good images of something either
good or bad, and lots of them. One of our first obligations to the customer
was making sure they could do their job, and meet their employers'
expectations. I always made sure that the people I was training were told of
the items you mentioned. I think that if you neglect educating the customer
about what all the options they have on their systems you are doing them a
disservice ("options" is equal to "detectors" in the case). You are not
fulfilling your contract/agreement with the customer to provide them with
the education they need to get the most out of the product you have sold to
them. It is important to remember that there are at the very least 2
detectors available, and they will provide different
images/data/perspectives of the same feature. It is a good idea to take
advantage of them. Often users are more focused on treating the SEM in
general as a camera (point and shoot) for getting an image of their
preconceived ideal/or not so ideal features/defects. Many users, not all,
(readers of this list are an exception) forget that they are using a tunable
microscope capable of scientific discovery. The SEM (FESEM) is dynamic
system. A system where when parameters are changed different information is
made visible and available. Learning about your SEM (FESEM) is a life long
journey.

To this idea I add the following. This morning I encountered one of my
newer/less experienced users on my FESEM in our clean room here at RPI, and
he complained that the images he was getting weren't as good as those his
company was getting from a contract lab. (He has less than 10 hours on the
system.) After spending some time with him, and showing him a few things,
and changing some parameters he said "I didn't think I could do this." I
think newer users are often driven to get lots of data/images quickly, and
they don't always remember or to take the time, to change a few things. They
don't try to see if they can make an image better with different
conditions/detectors. They are either afraid to take a bad picture, think
outside the box, or see what all different "knobs and buttons" do. Changing
parameters isn't going to break the machine. Today, images aren't costing 2
dollars a sheet. If the picture is bad, hit delete and move on and take a
new one.

The bottom line here is that I think the applications people at the major
microscope companies try and teach customers all they need to know. Do they
(the customers) hear and absorb it all? Not likely. We are all at fault in
this area that's why it is a good practice to take notes when learning about
new things. Education on any subject should be a life long endeavor. You
should never be satisfied with your skills and knowledge. One should always
be striving toward self betterment and the expansion of your knowledge and
skill. I hope my views (and they are mine alone), add a useful voice to the
topic you want to address.

Regards,

David

M. David Frey
Senior Application Engineer
Rensselaer Polytechnic Institute
Low Center for Industrial Inn.
Center for Integrated Electronics
110 8th Street
CII 4161 (Office)
CII 6015 (Packages and Mail)
Troy, NY 12180
518-276-3323 (office)
518-698-2288 (mobile)
-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: Thursday, January 14, 2010 9:41 AM
To: freym2-at-rpi.edu

Hi Listers

Pre Christmas I spent several months south of the Equator working with some
very good scientists on state of the art SEM. All of these instruments were
FEG, with both in and out of lens secondary electron detectors. Having
spent the last week writing up reports it suddenly struck me that the
techniques I introduced should have been part of the standard training for
this type of instrument.

I was one of the first people in the world to run a double detector
instrument when I worked on a pre production SS Series instrument at ISI.
My task was to look at all the options made available with a double detector
instrument, the advantages and disadvantages of the different detectors with
different types of specimen. With that information, as I gradually
developed an understanding of the generation of the signals I was receiving,
I was able to acquire exactly the signal mix which optimised the image for
the task in hand.

Over these past months it was very clear that the people I worked with just
knew that using the upper and short working distances gave them high
resolution, the subtleties of double detection instruments were unknown
because they had not been taught how to use the instrument correctly. I
think of the state of the art SEM as a racing car; my other life. There are
times, when it is easy to do so, that you go flat out because you have so
much potential, but there are other times when flat out makes the task much
more difficult. That is state of the art SEM, you have so much resolution
available but you really need other instrument attributes to obtain the
perfect image. So often I witnessed a quest for resolution when, with the
potential of the instrument, the chronic charging of the specimen should
have taken priority and I don't just mean turning down the accelerating
voltage.

So the point of this note, do people suffer from the same problems in the
northern hemisphere in that the subtleties of operation are ignored when
manufacturers train operators on state of the art FEG SEM? There are more
FEG SEM in the northern hemisphere so does that mean the knowledge of the
manufacturer's staff is of a higher level?

Thoughts?

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



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From: John.Mardinly-at-wdc.com
Date: Thu, 14 Jan 2010 10:57:15 -0600
Subject: [Microscopy] Re: Blind Student in Biology Lab

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

Don't forget Edward DeMille Campbell, one of the greatest researchers of
metallurgy. He was blinded by a laboratory explosion at age 28, but went on
to an extraordinary career of teaching and research. Today, the highest
award given annually by ASM is the Campbell lectureship. More at:
http://pubs.acs.org/doi/abs/10.1021/ie50191a048


John Mardinly
Western Digital

-----Original Message-----
X-from: schooley-at-mcn.org [mailto:schooley-at-mcn.org]
Sent: Wednesday, January 13, 2010 1:39 PM
To: John Mardinly

You've received a lot of clever, sensitive & helpful replies to your
inquiry. I'd like to suggest that you buy a book to share with your
lab staff (you can find it online for little more than the postage).
Geeraty Vermeij is arguably the world's leading authority on
molluscan morphological development, and he's been blind since early
childhood. He's on the U. of California at Davis faculty, and
"Priviledged Hands: a Scientific Life" is his autobiography.
--
Caroline Schooley
Project MICRO Coordinator
Microscopy Society of America
45301 Caspar Point Road, Box 117
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.microscopy.org/ProjectMICRO

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From: glenmac-at-u.washington.edu
Date: Thu, 14 Jan 2010 11:39:00 -0600
Subject: [Microscopy] PEN slides for microsdissection

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

Hello
A colleague is using our Leica microdissection system to isolate microglia for RNA extraction. The specimen is 5 um thick cryostat sections of paraformaldehyde fixed rat brain mounted on PEN film slides.

She needs to identify the cells to cut out by labeling with a biotinylated lectin but the the sections are coming off during the labeling steps.

Has anyone been able to overcome this problem by brand of PEN slides, subbing, prolonged drying...?

Thanks,
Glen


Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923 USA
(206) 616-4156
glenmac-at-uw.edu










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From: l.tilley-at-latrobe.edu.au
Date: Thu, 14 Jan 2010 13:47:26 -0600
Subject: [Microscopy] Postdoctoral position to work on High Resolution Imaging in

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

Debby,

Thanks for adding to what I have written. I often find here at RPI, also an
academic environment that the skills and knowledge a user brings into the
training that is provided helps them go much farther in the quality of the
images they produce. It also helps if they are interested in learning more
than steps 1 through 20 in how to get an image. Understanding the "basics"
of electron microscopy is a must.

X-from my prior experience as an applications person for a microscope company,
we often made many assumptions to the user's basic knowledge of electron
microscopy. As an aps person, we didn't always have time to fully educate
the customer on the "basics" of electron microscopy, we had 2 or 3 days to
get them to "know" their microscope, and then we off to home and our
families or on to the next customer.

I do like the fact that there is some requirement at Purdue for the users to
have a full credit course and lab behind them before they have full access
to the systems. Here at RPI there are a few people trying to push that sort
of requirement forward. I just always remind each user as they get trained
that they can always call, asking for help is good, and no question is a
stupid question.

One last idea I'd like to offer is that I take their science away from them
when teaching users to use the microscope. I make them look at my samples,
and they are focused on learning the basics, the controls, and the
microscope itself. They aren't asking why didn't my experiment work, they
are asking how do I make the picture better.

I offer that we should take the following conclusion away from this
discussion: Microscope vendors train users to user their products. They
aren't always to best source of basic electron microscopy education. (I
think the Norm Burns said this often at Cambridge/LEICA/LEO/Zeiss). As a
vendor we should leave it to the likes of Steve, Lehigh, or the old PASEM
course, or the New Paltz course at the Mountain House, or the classes
offered at our respective/favorite academic intuitions of choice. The
microscope vendors have a wonderful pool of knowledgeable people who are
tasked with selling and supporting a very technical product. At the end of
the day, month, or year the bottom line to microscope vendors is how many
did we sell, and how many happy customers do we have.*

David

* You can look at this as if it is driver's education, you don't see Ford,
or VW teaching driver's education, they sell cars. When you pick up your new
car they'll show you the basics, where the key goes, and such but it is
assumed you know the rules of the road and have driver's license. :)

M. David Frey
Senior Application Engineer
Rensselaer Polytechnic Institute
Low Center for Industrial Inn.
Center for Integrated Electronics
110 8th Street
CII 4161 (Office)
CII 6015 (Packages and Mail)
Troy, NY 12180
518-276-3323 (office)
518-698-2288 (mobile)

-----Original Message-----
X-from: Sherman, Debra M [mailto:dsherman-at-purdue.edu]
Sent: Thursday, January 14, 2010 1:25 PM
To: freym2-at-rpi.edu


} Postdoctoral position to work in the ARC Centre of Excellence for
} Coherent X-ray Science. (http://www.coecxs.org/)
}
} Research Officer
} La Trobe Institute for Molecular Science
} Level A/B Research Position in the Department of Biochemistry
} La Trobe Institute for Molecular Science
} This position will attract a remuneration package of approx. $70,200
} to $96,100 per annum, which includes 17% superannuation.
}
} Background:
} This position is funded by the Australian Research Council as part
} of the Centre of Excellence for Coherent X-ray Science (CXS) for
} research into the use of novel imaging techniques to study the
} cellular architecture of malaria parasite-infected erythrocytes. The
} CXS is an interdisciplinary collaboration for high-resolution
} bio-imaging. The project involves developing new methods and
} applying existing cutting edge techniques for light, electron and
} X-ray microscopy. The successful applicant will work with colleagues
} from the Departments of Biochemistry and Physics at La Trobe
} University. Molecular and cell biological manipulation of samples
} will be used to enhance specimen preparation.
}
} Primary Objectives:
} This work aims to develop sample preparation protocols suitable for
} high resolution imaging and to use these methods to obtain
} information about the cellular architecture of parasitised erythrocytes.
}
} In particular, we aim:
} (1) To develop sample preparation methods for transmission electron
} microscopy and cryo electron microscopy
} (2) To use electron tomography to image cell samples
} (2) To use X-ray microscopy to image samples
} (3) To use X-ray methods to examine the sub-cellular distribution
} and organisation of structures in malaria parasites and other samples
} (4) To use and develop super-resolution optical microscopy methods
}
} Further information,
} Contact: Prof. Leann Tilley. Tel: 03-9479 1375. Email:
} L.Tilley-at-latrobe.edu.au
} http://www.latrobe.edu.au/biochemistry/lab/tilley/index.htm
} Closing date for applications: mid February, 2010



Leann Tilley, Professor of Biochemistry
Director of Research, La Trobe Institute for Molecular Science
Deputy Director, ARC Centre of Excellence for Coherent X-ray Science

Department of Biochemistry Phone: 61-3-94791375
Rm 407, Phys Sci Bld 4 Fax: 61-3-94792467
Plenty Rd, La Trobe University Email: L.Tilley-at-latrobe.edu.au
Melbourne, 3086, Australia
http://www.latrobe.edu.au/biochemistry/lab/tilley/index.htm


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From: lee-at-asc.magnet.fsu.edu
Date: Thu, 14 Jan 2010 14:00:52 -0600
Subject: [Microscopy] TEM Laboratory Support Scientist Position Open at FSU

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

Job Title: Advanced Analytical TEM Laboratory Support Scientist
Job ID: 31065
Location: National High Magnetic Field Laboratory at Florida State
University, Tallahassee, FL

In conjunction with the recent purchase of a new JEM-ARM200F Atomic
Resolution Analytical TEM we have an opening for a TEM Laboratory Support
Scientist.

Qualifications: Advanced University degree(Master's or higher) and at
least four years of demonstrated experience.

Responsibilities:

*Interacting, assisting and training users for routine operation of the
TEM/STEM microscopes and related sample preparation equipment.
*Performing routine maintenance and essential repair of the microscopes
and related sample preparation equipment (major equipment is under service
contract).
*Managing routine operation, including logs, bookkeeping, billing,
maintenance records, inventory, and consumables purchases.
*Making recommendations and assist in the purchase of new equipment.
*Participating in the microscopy community and related research
activities.
*Preparing monthly and annual reports concerning the TEM Laboratory and
participate in all required review meetings.
*Providing outreach and tours for the general public.

More information and the on-line application (Job ID 31065) is available
at https://jobs.fsu.edu
The NHMFL web site can be found at: http://www.magnet.fsu.edu/
A partial listing of existing analytical facilities at NHMFL can be found
at:
http://www.magnet.fsu.edu/magnettechnology/research/materials/microanalysi
s/index.html

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From: m_raven-at-lifesci.ucsb.edu
Date: Thu, 14 Jan 2010 15:50:25 -0600
Subject: [Microscopy] Workshops/Training for TEM with Biological Samples

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

Hi Fred

I agree with all that you mention and I think it is important as a vendor to
work hard on judging your customer.

If I run a basic residential course very few wish to attend, but run an
advanced course and that creates interest. However when I start this course
from the same data that is in the basic course the "advanced" users are
fascinated because they too did not know! What I think we may be saying is
that not all customers are up to speed on SEM in general and therefore it is
tough for them to understand a State of the Art instrument.

But back to my first email. When you sell a State of the Art piece of
equipment you owe it to your client to bring them up to State of the Art or
in my mind what is the point of selling the equipment? All of us who have
been in the business know that the more you sell, the more you sell. With
that aim the better trained your new customer is the more papers they will
write and the good name of your product is enhanced. That was the
philosophy with which I had always worked and operating as I do now, often
through personal recommendation, I agree it is important to "do a good job".


Lets await more comment

Steve



-----Original Message-----
X-from: freym2-at-rpi.edu [mailto:freym2-at-rpi.edu]
Sent: 14 January 2010 19:33
To: protrain-at-emcourses.com

Debby,

Thanks for adding to what I have written. I often find here at RPI, also an
academic environment that the skills and knowledge a user brings into the
training that is provided helps them go much farther in the quality of the
images they produce. It also helps if they are interested in learning more
than steps 1 through 20 in how to get an image. Understanding the "basics"
of electron microscopy is a must.

X-from my prior experience as an applications person for a microscope
company,
we often made many assumptions to the user's basic knowledge of electron
microscopy. As an aps person, we didn't always have time to fully educate
the customer on the "basics" of electron microscopy, we had 2 or 3 days to
get them to "know" their microscope, and then we off to home and our
families or on to the next customer.

I do like the fact that there is some requirement at Purdue for the users to
have a full credit course and lab behind them before they have full access
to the systems. Here at RPI there are a few people trying to push that sort
of requirement forward. I just always remind each user as they get trained
that they can always call, asking for help is good, and no question is a
stupid question.

One last idea I'd like to offer is that I take their science away from them
when teaching users to use the microscope. I make them look at my samples,
and they are focused on learning the basics, the controls, and the
microscope itself. They aren't asking why didn't my experiment work, they
are asking how do I make the picture better.

I offer that we should take the following conclusion away from this
discussion: Microscope vendors train users to user their products. They
aren't always to best source of basic electron microscopy education. (I
think the Norm Burns said this often at Cambridge/LEICA/LEO/Zeiss). As a
vendor we should leave it to the likes of Steve, Lehigh, or the old PASEM
course, or the New Paltz course at the Mountain House, or the classes
offered at our respective/favorite academic intuitions of choice. The
microscope vendors have a wonderful pool of knowledgeable people who are
tasked with selling and supporting a very technical product. At the end of
the day, month, or year the bottom line to microscope vendors is how many
did we sell, and how many happy customers do we have.*

David

* You can look at this as if it is driver's education, you don't see Ford,
or VW teaching driver's education, they sell cars. When you pick up your new
car they'll show you the basics, where the key goes, and such but it is
assumed you know the rules of the road and have driver's license. :)

M. David Frey
Senior Application Engineer
Rensselaer Polytechnic Institute
Low Center for Industrial Inn.
Center for Integrated Electronics
110 8th Street
CII 4161 (Office)
CII 6015 (Packages and Mail)
Troy, NY 12180
518-276-3323 (office)
518-698-2288 (mobile)

-----Original Message-----
X-from: Sherman, Debra M [mailto:dsherman-at-purdue.edu]
Sent: Thursday, January 14, 2010 1:25 PM
To: freym2-at-rpi.edu

Dear All
Could you direct me to short courses, ideally hands-on, that focus on
biological sample preparation for TEM and TEM imaging of Biological
Samples? I'd like to generate a list for my facility.
Best Regards
Mary

--
Mary Raven
Integrated Microscopy Facility Director
Neuroscience Research Institute &
Molecular, Cellular & Developmental Biology
The University of California
Santa Barbara, CA 93106-5060

https://ucsb-microscopy.ucsd.edu/schedule/
http://www.lifesci.ucsb.edu/mcdb/research/facilities/microscopy/microscopy.html

Phone: (805) 893 8702
Fax: (805) 893 2005



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From: TindallR-at-missouri.edu
Date: Thu, 14 Jan 2010 15:56:33 -0600
Subject: [Microscopy] Workshops/Training for TEM with Biological Samples

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

Hi Mary,

We will provide custom training in-house on your samples in any area of EM we have the equipment to handle. We have done intensive 1-2 week sessions with individuals in the past and also will work with small groups, if needed. We pretty much tailor everything to the client's personal needs, so costs highly variable.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




-----Original Message-----
X-from: m_raven-at-lifesci.ucsb.edu [mailto:m_raven-at-lifesci.ucsb.edu]
Sent: Thursday, January 14, 2010 3:51 PM
To: Tindall, Randy D.

Dear All
Could you direct me to short courses, ideally hands-on, that focus on
biological sample preparation for TEM and TEM imaging of Biological
Samples? I'd like to generate a list for my facility.
Best Regards
Mary

--
Mary Raven
Integrated Microscopy Facility Director
Neuroscience Research Institute &
Molecular, Cellular & Developmental Biology
The University of California
Santa Barbara, CA 93106-5060

https://ucsb-microscopy.ucsd.edu/schedule/
http://www.lifesci.ucsb.edu/mcdb/research/facilities/microscopy/microscopy.html

Phone: (805) 893 8702
Fax: (805) 893 2005



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From: dsherman-at-purdue.edu
Date: Thu, 14 Jan 2010 18:14:32 -0600
Subject: [Microscopy] State of the Art SEM

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

I think David is right on as to the expectations of many users. Many are
constantly under time-pressure and will work to provide the best they can in
the limited time available and just do not have time to try lots of
variables to get the best possible image. I do not condone this but do
understand it.

I work in an academic environment where we have lots of researchers (mainly
graduate students) who treat EM as an occasional technique to supplement to
their primary research tools. This is fine to a point. If the images are
poor than the reflect poorly on the student, the major professor, the lab
where they were generated (me!!), and the institution.

We require students take a for-credit graduate-level course on TEM (or SEM)
theory and the hands-on lab in order to have the "privilege" for independent
access to the instruments. This gets through the first step of making sure
all users have the basic information to make informed decisions about
imaging parameters, sample preparation, and an introduction to the ethics of
image processing, interpretation, etc. However, once the course is over and
the students are on their own, it is their choice as to how careful they are
in acquiring the best possible images and correctly interpreting those
images.

Some are very good about asking for help and they are the ones who normally
will do well and produce excellent images consistently. Some are naturals
who absorb all they are taught and just have a knack for good imaging. Then
there are those who never ask for help, know it all, and only want to work
in the middle of the night when there is no one around for help. Those are
the ones who often do not do as well.

This goes back to the recent string on our responsibility for data. It is
out of our hands and we can only hope that we have laid down a solid
foundation of knowledge through formal courses and follow-up when asked.
Then it is up to the microscopist as to if or how they use that knowledge.

Debby

--
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy/


} From: "freym2-at-rpi.edu" {freym2-at-rpi.edu}
} Reply-To: "freym2-at-rpi.edu" {freym2-at-rpi.edu}
} Date: Thu, 14 Jan 2010 11:55:31 -0500
} To: Debby Sherman {dsherman-at-purdue.edu}
} Subject: [Microscopy] RE: State of the Art SEM
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi Steve,
}
} It is a very interesting point you bring up. I spent nearly 10 years working
} for one the manufactures of High res- FESEM's as an applications specialist.
} The topics/concepts that were emphasized in training were often driven by
} the demands of the customer. This seemed to be more true in an onsite
} training rather than those we would provide to a mixed group (people from
} different companies) at our offices/demo labs. This maybe right or wrong
} depending on where you stand on the following statements, "The customer
} doesn't know either what he really wants, or what he really needs." And the
} "Customer is always right." I think that it is often the case that the
} applications people are being driven by trying to meet the customers
} demands/expectations, "I purchased a HI-RES FESEM, I demand HI-RES 100% of
} the time." Many of the FESEM's that we sold were into an "industrial"
} environment where good and knowledgeable people were always under the gun,
} and had an end result they needed to meet. Good images of something either
} good or bad, and lots of them. One of our first obligations to the customer
} was making sure they could do their job, and meet their employers'
} expectations. I always made sure that the people I was training were told of
} the items you mentioned. I think that if you neglect educating the customer
} about what all the options they have on their systems you are doing them a
} disservice ("options" is equal to "detectors" in the case). You are not
} fulfilling your contract/agreement with the customer to provide them with
} the education they need to get the most out of the product you have sold to
} them. It is important to remember that there are at the very least 2
} detectors available, and they will provide different
} images/data/perspectives of the same feature. It is a good idea to take
} advantage of them. Often users are more focused on treating the SEM in
} general as a camera (point and shoot) for getting an image of their
} preconceived ideal/or not so ideal features/defects. Many users, not all,
} (readers of this list are an exception) forget that they are using a tunable
} microscope capable of scientific discovery. The SEM (FESEM) is dynamic
} system. A system where when parameters are changed different information is
} made visible and available. Learning about your SEM (FESEM) is a life long
} journey.
}
} To this idea I add the following. This morning I encountered one of my
} newer/less experienced users on my FESEM in our clean room here at RPI, and
} he complained that the images he was getting weren't as good as those his
} company was getting from a contract lab. (He has less than 10 hours on the
} system.) After spending some time with him, and showing him a few things,
} and changing some parameters he said "I didn't think I could do this." I
} think newer users are often driven to get lots of data/images quickly, and
} they don't always remember or to take the time, to change a few things. They
} don't try to see if they can make an image better with different
} conditions/detectors. They are either afraid to take a bad picture, think
} outside the box, or see what all different "knobs and buttons" do. Changing
} parameters isn't going to break the machine. Today, images aren't costing 2
} dollars a sheet. If the picture is bad, hit delete and move on and take a
} new one.
}
} The bottom line here is that I think the applications people at the major
} microscope companies try and teach customers all they need to know. Do they
} (the customers) hear and absorb it all? Not likely. We are all at fault in
} this area that's why it is a good practice to take notes when learning about
} new things. Education on any subject should be a life long endeavor. You
} should never be satisfied with your skills and knowledge. One should always
} be striving toward self betterment and the expansion of your knowledge and
} skill. I hope my views (and they are mine alone), add a useful voice to the
} topic you want to address.
}
} Regards,
}
} David
}
} M. David Frey
} Senior Application Engineer
} Rensselaer Polytechnic Institute
} Low Center for Industrial Inn.
} Center for Integrated Electronics
} 110 8th Street
} CII 4161 (Office)
} CII 6015 (Packages and Mail)
} Troy, NY 12180
} 518-276-3323 (office)
} 518-698-2288 (mobile)
} -----Original Message-----
} X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
} Sent: Thursday, January 14, 2010 9:41 AM
} To: freym2-at-rpi.edu
} Subject: [Microscopy] State of the Art SEM
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi Listers
}
} Pre Christmas I spent several months south of the Equator working with some
} very good scientists on state of the art SEM. All of these instruments were
} FEG, with both in and out of lens secondary electron detectors. Having
} spent the last week writing up reports it suddenly struck me that the
} techniques I introduced should have been part of the standard training for
} this type of instrument.
}
} I was one of the first people in the world to run a double detector
} instrument when I worked on a pre production SS Series instrument at ISI.
} My task was to look at all the options made available with a double detector
} instrument, the advantages and disadvantages of the different detectors with
} different types of specimen. With that information, as I gradually
} developed an understanding of the generation of the signals I was receiving,
} I was able to acquire exactly the signal mix which optimised the image for
} the task in hand.
}
} Over these past months it was very clear that the people I worked with just
} knew that using the upper and short working distances gave them high
} resolution, the subtleties of double detection instruments were unknown
} because they had not been taught how to use the instrument correctly. I
} think of the state of the art SEM as a racing car; my other life. There are
} times, when it is easy to do so, that you go flat out because you have so
} much potential, but there are other times when flat out makes the task much
} more difficult. That is state of the art SEM, you have so much resolution
} available but you really need other instrument attributes to obtain the
} perfect image. So often I witnessed a quest for resolution when, with the
} potential of the instrument, the chronic charging of the specimen should
} have taken priority and I don't just mean turning down the accelerating
} voltage.
}
} So the point of this note, do people suffer from the same problems in the
} northern hemisphere in that the subtleties of operation are ignored when
} manufacturers train operators on state of the art FEG SEM? There are more
} FEG SEM in the northern hemisphere so does that mean the knowledge of the
} manufacturer's staff is of a higher level?
}
} Thoughts?
}
} Steve Chapman FRMS
} Senior Consultant Protrain
} For consultancy and training in electron microscopy world wide
} Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
} www.emcourses.com
}
}
}
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} 9, 27 -- Subject: State of the Art SEM
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} 23, 26 -- From freym2-at-rpi.edu Thu Jan 14 10:53:44 2010
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From: max_histo_00-at-yahoo.it
Date: Fri, 15 Jan 2010 08:06:56 -0600
Subject: [Microscopy] viaWWW: Used microtome

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Email: max_histo_00-at-yahoo.it
Name: Massimo Tosi

Organization: Private

Title-Subject: [Filtered] Used microtome

Message: Hi all,

I am an amateur naturalist, and I'm looking for an used microtome,
but working, even on old model, to be used for histological sections
of samples included in paraffin.
Please can anyone give me useful directions where to find it?

Thank you all for you help,

Massimo Tosi


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From: ZZhang-at-uwyo.edu
Date: Fri, 15 Jan 2010 09:46:02 -0600
Subject: [Microscopy] old diamond knives

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

Hi all:

I inherited several old diamond knives from a faculty who is retiring. The knives include one from SAG International (Italy), one Sakura Sapphatome (Japan) and a couple from Dupon. My question is:

Is there a way I can check to see which one is sharp and still good to use?

Thank you,

Zhaojie

Zhaojie Zhang, Ph.D.
Director, Microscopy Core Facility
Department of Zoology and Physiology
University of Wyoming
Laramie, WY 82071
zzhang-at-uwyo.edu
307-766-3038




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From: ZZhang-at-uwyo.edu
Date: Fri, 15 Jan 2010 10:54:45 -0600
Subject: [Microscopy] old diamond knives

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

Thank you all for your quick responses.

I've received several responses within minutes of my posting, suggesting the same thing - try to section with the old knife.

I guess I was hoping to find an "easier and quicker" way for this, like a magic wand. It apparently does not exist (!?).

Thank you all, again.

Zhaojie

-----Original Message-----
X-from: Haller, Edward [mailto:ehaller-at-health.usf.edu]
Sent: Friday, January 15, 2010 9:16 AM
To: Z.J. Zhang

Hi all:

I inherited several old diamond knives from a faculty who is retiring. The knives include one from SAG International (Italy), one Sakura Sapphatome (Japan) and a couple from Dupon. My question is:

Is there a way I can check to see which one is sharp and still good to use?

Thank you,

Zhaojie

Zhaojie Zhang, Ph.D.
Director, Microscopy Core Facility
Department of Zoology and Physiology
University of Wyoming
Laramie, WY 82071
zzhang-at-uwyo.edu
307-766-3038




==============================Original Headers==============================
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From: oshel1pe-at-cmich.edu
Date: Fri, 15 Jan 2010 15:20:27 -0600
Subject: [Microscopy] Fwd: FW: Ask-A-Microscopist

Contents Retrieved from Microscopy Listserver Archives
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HI STEVE,

I think in my first post you missed the part where I wrote that as a vendor
we made sure that the customer was shown all of the options/detectors on
their new systems. We would have failed at our jobs and our commitment to
the customer if we did not show them these items. As a provider of a complex
system such as a SEM, we had one focus and that focus was to make sure the
customer got the best out of their new microscope. We were not there to
teach the basics of electron microscopy (EM). We did, and do judge a
customer's level of proficiency, but generally the extent of our basic EM
educational responsibilities came to making sure the customer understands
the most basic relationships one needs to grasp in order to get the most out
of their new microscope. In a 2-3 day training with new a digital windows
based SEM control system there are so many different items to teach the user
about, spending a half a day to 1 day on the basic foundations of EM would
rob the customer of seeing entire feature sets that make the SEM a solution.
As a vendor we needed to help solve their problems, answer their questions,
make their work flow better. We did spend a great deal of time on the
detectors and how to get the best images from each of them. We also needed
to show them automation SW, image processing, stage navigation, scripting
sw, interfaces to external systems, and the list continues...You can not
learn in 2 days or even 5 days all there is to learn about EM, it is a
lifetime vocation or dare I even say devotion. Some people have been at it
their entire lives, and still learn new things every single day. Isn't that
what attracts us to this field, is the fact that there is more to learn then
we might ever learn in our entire lives...

So if we boil your initial post down to the following statement/question "Do
microscope vendors take the time to show users all the detectors and how to
get the best out of them?" The answer would be an unequivocal "YES!" We (and
I should write "They", since I am no longer working for one of them.)
wouldn't stay in business if we (they) didn't. Just saying "yes" is a rather
boring answer that does not create discussion amongst our peers as to the
best way to educate ourselves and those that we are charged with educating
about how to get the most out of your NEW STATE OF THE ART SEM. I guess we
need to pay people to come and show us what we can't figure out through
reading a manual, or a book, by flipping the "switches" and changing
parameters, or maybe by even calling the people that sold you the system and
saying, "the image from my lower detector is terrible, how do I make it
better, I can't figure it out. I need your help." For either the cost of a
phone call or the time to compose an email and attach a file to the sales
rep who sold the system, you will get the help you need. The sales rep will
help to get you in touch with someone who will answer your question or help
to solve your problem. (Veeco was great about this with the 4 year old AFM I
inherited when I got here, knowing nothing about AFM. Now I am competent at
AFM, and still learning everyday.) You could also use this wonderful list
server to get advice and guidance from your peers.

The reality in the business of technical education is if people don't seek
out constant growth and knowledge expansion, their skills will never
improve. You should never be satisfied with your level of skill, and
knowledge. Once you stop learning and discovering it is time to move on and
do something new.

I think that many customers (and I mean no offense to anyone in writing
this), get the basic orientation from their service rep to what things do on
their shiny NEW STATE OF THE ART SEM, and they think my skill level, and
education is good enough to do the rest. For some this is true for others
they might miss out on a few things. Often customers don't want to, or can't
(more times than not it is can't) spend the money to travel to the
demonstration laboratories, of the microscope companies, so they miss out on
the initial user trainings where for the cost of a plane ticket, and few
nights in a hotel, learn all the things they already know and some new
things they need to know about their shiny new SEM. If you think about the
amount of money they have just spent on their new microscope, the cost to
attend the training or get an on-site training is a small fraction of what
they have just spent. There were many occasions when we sent introduction
letters to customers that gave them the dates for the in-house training
classes, and never got any response. We would only hear from that customer
if they were unhappy with the performance of their new system.

Even if you are the best at something someone always has something new they
can teach you. Even it is something as simple as the joy of discovery, or
another way of looking at things or sharing your knowledge to enrich the
person teaching you. I would suggest to anyone who has recently purchased a
NEW STATE OF THE ART SEM, call the people who sold it to you and ask them
for training class dates. You will learn a lot from the people who built and
sold you your SEM, even if you have 10,000 hours or more experience. They
are in the best position to teach you about their systems and their
technology. If it is an e cross b filter or a esb detector or whatever is
latest blip of technology. They know it, live it and built it. You may not
learn all there is know about EM, (go to Lehigh or the like for your local)
but you will learn all the ins and outs of your microscope. Some vendors
even offer free training for life depending upon from whom you purchased
your system.

Best of luck, happy imaging and keep on learning....

"FRED" aka David Frey


PS- I think that Debby closed with a great point that once we have
trained/educated people on "our" microscopes we can't always control how or
if they use the education they have received. The quality of images they are
satisfied producing it is out of our hands. We here at RPI have over 75
users qualified to user our FESEM. I can't, and am not given the opportunity
to review all the images that are produced. We can only hope that people ask
for help if they aren't satisfied with their images. -MDF


M. David Frey
Senior Application Engineer
Rensselaer Polytechnic Institute
Low Center for Industrial Inn.
Center for Integrated Electronics
110 8th Street
CII 4161 (Office)
CII 6015 (Packages and Mail)
Troy, NY 12180
518-276-3323 (office)
518-698-2288 (mobile)
-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: Thursday, January 14, 2010 4:15 PM
To: freym2-at-rpi.edu

Hi Fred

I agree with all that you mention and I think it is important as a vendor to
work hard on judging your customer.

If I run a basic residential course very few wish to attend, but run an
advanced course and that creates interest. However when I start this course
from the same data that is i the basic course the "advanced" users are
fascinated because they too did not know! What I think we may be saying is
that not all customers are up to speed on SEM in general and therefore it is
tough for them to understand a State of the Art instrument.

But back to my first email. When you sell a State of the Art piece of
equipment you owe it to your client to bring them up to State of the Art or
in my mind what is the point of selling the equipment? All of us who have
been in the business know that the more you sell, the more you sell. With
that aim the better trained your new customer is the more papers they will
write and the good name of your product is enhanced. That was the
philosophy with which I had always worked and operating as I do now, often
through personal recommendation, I agree it is important to "do a good job".


Lets await more comment

Steve



-----Original Message-----
X-from: freym2-at-rpi.edu [mailto:freym2-at-rpi.edu]
Sent: 14 January 2010 19:33
To: protrain-at-emcourses.com

Debby,

Thanks for adding to what I have written. I often find here at RPI, also an
academic environment that the skills and knowledge a user brings into the
training that is provided helps them go much farther in the quality of the
images they produce. It also helps if they are interested in learning more
than steps 1 through 20 in how to get an image. Understanding the "basics"
of electron microscopy is a must.

X-from my prior experience as an applications person for a microscope
company,
we often made many assumptions to the user's basic knowledge of electron
microscopy. As an aps person, we didn't always have time to fully educate
the customer on the "basics" of electron microscopy, we had 2 or 3 days to
get them to "know" their microscope, and then we off to home and our
families or on to the next customer.

I do like the fact that there is some requirement at Purdue for the users to
have a full credit course and lab behind them before they have full access
to the systems. Here at RPI there are a few people trying to push that sort
of requirement forward. I just always remind each user as they get trained
that they can always call, asking for help is good, and no question is a
stupid question.

One last idea I'd like to offer is that I take their science away from them
when teaching users to use the microscope. I make them look at my samples,
and they are focused on learning the basics, the controls, and the
microscope itself. They aren't asking why didn't my experiment work, they
are asking how do I make the picture better.

I offer that we should take the following conclusion away from this
discussion: Microscope vendors train users to user their products. They
aren't always to best source of basic electron microscopy education. (I
think the Norm Burns said this often at Cambridge/LEICA/LEO/Zeiss). As a
vendor we should leave it to the likes of Steve, Lehigh, or the old PASEM
course, or the New Paltz course at the Mountain House, or the classes
offered at our respective/favorite academic intuitions of choice. The
microscope vendors have a wonderful pool of knowledgeable people who are
tasked with selling and supporting a very technical product. At the end of
the day, month, or year the bottom line to microscope vendors is how many
did we sell, and how many happy customers do we have.*

David

* You can look at this as if it is driver's education, you don't see Ford,
or VW teaching driver's education, they sell cars. When you pick up your new
car they'll show you the basics, where the key goes, and such but it is
assumed you know the rules of the road and have driver's license. :)

M. David Frey
Senior Application Engineer
Rensselaer Polytechnic Institute
Low Center for Industrial Inn.
Center for Integrated Electronics
110 8th Street
CII 4161 (Office)
CII 6015 (Packages and Mail)
Troy, NY 12180
518-276-3323 (office)
518-698-2288 (mobile)

-----Original Message-----
X-from: Sherman, Debra M [mailto:dsherman-at-purdue.edu]
Sent: Thursday, January 14, 2010 1:25 PM
To: freym2-at-rpi.edu

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of
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==========================================================

} Subject: FW: Ask-A-Microscopist
} Date: Fri, 15 Jan 2010 16:03:48 -0500
} From: "Roberta Omachel" {romachel-at-DROHANMGMT.COM}
} To: {oshel1pe-at-cmich.edu}
}
} -----Original Message-----
} From: Joon Yang [mailto:jhyang01-at-gmail.com]
} Sent: Tuesday, November 03, 2009 3:53 PM
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Tuesday, November 03,
} 2009 at 12:52:28 PM.
}
} realname - Joon Yang
} Email - jhyang01-at-gmail.com
} ORGANIZATION - University of Maryland
} EDUCATION - Graduate College
} SUBJECT_OF_QUESTION - convergent beam diffraction (CBD) ?
} QUESTION - Hi, I would like to analyze convergent beam diffraction (CBD)
} of my thin film samples. I like to compare it to the simulation result
} of CBD. However, I could not find CBD function in EMS website. Is there
} any web site or free software? I can not afford to buy the sofware.
} Please help me and let me know.
}
} Sincerely,
}
} Joon
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: oshel1pe-at-cmich.edu
Date: Fri, 15 Jan 2010 15:23:12 -0600
Subject: [Microscopy] HIgh school with SEM looking for research education

Contents Retrieved from Microscopy Listserver Archives
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==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of
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list.
==========================================================

} Subject: FW: Ask-A-Microscopist
} -----Original Message-----
} From: Mrs. Heather Fogell [mailto:Fogellh-at-rlasd.k12.pa.us]
} Sent: Thursday, November 05, 2009 9:24 PM
} To: AssociationManagement
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Thursday, November
} 05, 2009 at 06:23:53 PM.
}
} realname - Mrs. Heather Fogell
} Email - Fogellh-at-rlasd.k12.pa.us
} ORGANIZATION - Red Lion Area Senior High School
} EDUCATION - 9-12th Grade High School
} LOCATION - Red Lion, PA 17356
} SUBJECT_OF_QUESTION - Biology
} QUESTION - I am a teacher at the Red Lion Senior HS in Red Lion,
} Pa., who is responsible for an SEM (EVAX) program within the High
} School. The SEM has generated a great deal of interest and the
} students are excited about real world applications. I would like to
} broaden the high school student's experience by teaming up with some
} undergrad or grad program and trade the use of our SEM in exchange
} for allowing my students a "window" on the scientific research. My
} students could even do some of the technician work. I want allow for
} my students to make the connection between the complex equipment and
} the "real world" research applications while having them experience
} the scientific process. If interested, or if you have any ideas or
} suggestions, please feel free to contact me.
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: oshel1pe-at-cmich.edu
Date: Fri, 15 Jan 2010 15:50:52 -0600
Subject: [Microscopy] Fwd: FW: Ask-A-Microscopist

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

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of
MSA, and any reply should go directly to the poster as well as to the
list.
==========================================================

} Subject: FW: Ask-A-Microscopist
} Date: Fri, 15 Jan 2010 16:04:41 -0500
}
} -----Original Message-----
} From: Eric Coates [mailto:ericcoates-at-hotmail.com]
} Sent: Tuesday, December 01, 2009 1:35 PM
} To: AssociationManagement
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Tuesday, December 01,
} 2009 at 10:35:14 AM.
}
} realname - Eric Coates
} Email - ericcoates-at-hotmail.com
} SUBJECT_OF_QUESTION - freeze substitution
} QUESTION - I am looking for a way to perform freeze substitution of ice
} crystals for optical, not SEM, analysis. I am familiar with one method,
} described in the article "Low Temperature SEM of Precipitated and
} Metamorphosed Snow Crystals Collected and Transported from Remote Sites"
} that appeared in JAMA v2 n3 1996, but it involves osmium tetroxide,
} which I would like to avoid. I have also tried silver nitrate with no
} success. Any suggestions would be appreciated.
}
} Thanks
}
} Eric Coates
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: jflaci-at-ms.sapientia.ro
Date: Sat, 16 Jan 2010 16:03:24 -0600
Subject: [Microscopy] Choice for coating teeth structures for SEM ...

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

Hello everybody!

I have just started to work an a SEM, which was acquired by donation at my
university, and I started to bump into questions. Maybe You could point to
me a good source of information, I am willing to learn. I have experience about
3 years in TEM, but this is a way other thing.

I am having trouble choosing the right coating for this work. I have
to image the surface
of tooth and the interior tooth canal. I tried to work with a 2nm
coating of gold, which lead
to bit of charge up, and I have seen in some article that others are
working with 20nm.

If everybody could help, what thickness would be the right choice for
this kind of work.
My sem is an old JEOL JSM-5200.


Jakab-Farkas Laszlo
Laboratory technician

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From: jflaci-at-ms.sapientia.ro
Date: Sat, 16 Jan 2010 17:11:40 -0600
Subject: [Microscopy] Re: Choice for coating teeth structures for SEM ...

Contents Retrieved from Microscopy Listserver Archives
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Hello, and thanks for both of You for the quick answers. So as I can now see,
the 20nm of coating could the proper one. Carbon coating possibility
is currently not available for me, becouse the evaporator is set up
for gold now, but I will try that
also in the very near future. Of course I am splitting them, I can not
imagine how to
to this other way.

JAKAB-FARKAS Laszlo

2010/1/17 Ron L'Herault {lherault-at-bu.edu} :
} OUr SEM is uses a tungsten filament and we only image between 10x and 5000x,
} and I have looked at a lot of teeth, both from the inside and outside.  I
} started on a JEOL JSM 35U, although we now use a Philips XL 20.  Our old
} Technics sputter coater puts down about 200 Angstroms of gold/palladium or
} pure gold, depending on the target I install.  Straight gold is not as good
} as the mix for rough surfaces because the mix makes smaller grains.  I
} imagine you are splitting the teeth open to reveal the canals, right?   It
} is very difficult (maybe impossible) to get coatings down inside of a long
} tube.
}
} Ron L'Herault
} Biomaterials Division
} Boston University
} Goldman School of Dental Medicine
}
} -----Original Message-----
} From: jflaci-at-ms.sapientia.ro [mailto:jflaci-at-ms.sapientia.ro]
} Sent: Saturday, January 16, 2010 5:08 PM
} To: lherault-at-bu.edu
} Subject: [Microscopy] Choice for coating teeth structures for SEM ...
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} Hello everybody!
}
} I have just started to work an a SEM, which was acquired by donation at my
} university, and I started to bump into questions. Maybe You could point to
} me a good source of information, I am willing to learn. I have experience
} about
} 3 years in TEM, but this is a way other thing.
}
} I am having trouble choosing the right coating for this work. I have
} to image the surface
} of tooth and the interior tooth canal. I tried to work with a 2nm
} coating of gold, which lead
} to bit of charge up, and I have seen in some article that others are
} working with 20nm.
}
} If everybody could help, what  thickness would be the right choice for
} this kind of work.
} My sem is an old JEOL JSM-5200.
}
}
} Jakab-Farkas Laszlo
} Laboratory technician
}
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From: gary-at-gaugler.com
Date: Sat, 16 Jan 2010 17:13:16 -0600
Subject: [Microscopy] Re: Choice for coating teeth structures for SEM

Contents Retrieved from Microscopy Listserver Archives
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Au is not my choice for anything being coated. It produces
a spider web characteristic if not done at high vacuum.
Try Au/Pd or Ir at 30mT and don't worry all that much
about film thickness. 50nm or thereabouts should work fine.

gary g.


At 02:05 PM 1/16/2010, you wrote:



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From: jflaci-at-ms.sapientia.ro
Date: Sat, 16 Jan 2010 17:21:30 -0600
Subject: [Microscopy] Re: Choice for coating teeth structures for SEM ...

Contents Retrieved from Microscopy Listserver Archives
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Hello!

Sorry for not observing Your message, the voltage would be 5 and 25 kV, becouse
as I have observed on the first look on the samples, the high
accelerating voltage
reveals also some of the underlying structure. As I have decreased the
voltage, some of the features disappeared, and i reached the
conclusion that it happened because the lower accelerating voltage /
lower energy electrons have not penetrated the sample as the 25kV
electrons did. Or was I wrong in this thinking process?

About the beam size: this type of instrument has a beam size setting
without the
possibility to see the real beam size, but I have been working at low
magnification
with higher, and as I increased the magnification I decreased the beam size.
It has a setting trough a potentiometer control from 7o' clock to 5o' clock.
for 500-1000x i use a setting about 2-3 o' clock, and when I go higher
in magnification i use a setting on a low as possible basis around
9-12 o' clock.

Thanks for Your help,
Laci

2010/1/17 Markus F. Meyenhofer {micro-at-superlink.net} :
} What KV?, Beam size?
} markus
} ----- Original Message ----- From: {jflaci-at-ms.sapientia.ro}
} To: {micro-at-superlink.net}
} Sent: Saturday, January 16, 2010 5:03 PM
} Subject: [Microscopy] Choice for coating teeth structures for SEM ...
}
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} } Hello everybody!
} }
} } I have just started to work an a SEM, which was acquired by donation at my
} } university, and I started to bump into questions. Maybe You could point to
} } me a good source of information, I am willing to learn. I have experience
} } about
} } 3 years in TEM, but this is a way other thing.
} }
} } I am having trouble choosing the right coating for this work. I have
} } to image the surface
} } of tooth and the interior tooth canal. I tried to work with a 2nm
} } coating of gold, which lead
} } to bit of charge up, and I have seen in some article that others are
} } working with 20nm.
} }
} } If everybody could help, what  thickness would be the right choice for
} } this kind of work.
} } My sem is an old JEOL JSM-5200.
} }
} }
} } Jakab-Farkas Laszlo
} } Laboratory technician
} }
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--
Jakab-Farkas Laszlo
Labortechnikus

SAPIENTIA - ERDÉLYI MAGYAR TUDOMÁNYEGYETEM
MÜSZAKI ÉS HUMÁNTUDOMÁNYOK KAR MAROSVÁSÁRHELY
Campus: Segesvári út 1C., Marosvásárhely/Koronka (a város határában)
Tel.: +40-(0)265-206210,+40-(0)265-208170, fax: 0265-206211
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E-mail: jflaci-at-ms.sapientia.ro
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From: jflaci-at-ms.sapientia.ro
Date: Sat, 16 Jan 2010 17:25:27 -0600
Subject: [Microscopy] Re: Choice for coating teeth structures for SEM

Contents Retrieved from Microscopy Listserver Archives
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Hello,

I make the coating around 0.003 to 0.006 mTorr.
Does this effect of spider web appear at this pressure?


Laci

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From: gary-at-gaugler.com
Date: Sat, 16 Jan 2010 17:41:07 -0600
Subject: [Microscopy] Choice for coating teeth structures for

Contents Retrieved from Microscopy Listserver Archives
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If you want surface features, you use low KV. High KV
will have too much volumetric interaction and lose surface
detail.

At low mag, I don't see much issue with probe diameter.
However, probe current would be an issue as is KV.
It depends on how much you want to collect below the surface.

I don't recommend C for surface detail...too coarse.
Use a metal. Good ones are Au/Pd, Pd and Ir in a cold
sputter coater. Chrome evaporation works well for a few
minutes but oxidizes quickly. The others will last
indefinitely.

gary g.


At 03:23 PM 1/16/2010, you wrote:

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From: jflaci-at-ms.sapientia.ro
Date: Sat, 16 Jan 2010 17:46:33 -0600
Subject: [Microscopy] Choice for coating teeth structures for SEM

Contents Retrieved from Microscopy Listserver Archives
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I use an oil diffusion pump and a rotation pump.
I measure the pressure with an ionization gauge,
and I get a reading about 3-6 x 10 -6 Torr.

By mT You ment mili Torr, or was I confused about that?

Laci

2010/1/17 Gary Gaugler {gary-at-gaugler.com} :
} How can you get to 0.003-0.006mT?  If real, that
} is too high of vacuum.  For ultra fine coating, I
} work at 15mT with turbo pumped coater.
}
} gary g.
}
}
} At 03:26 PM 1/16/2010, you wrote:
}
}
}
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
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} }
} } Hello,
} }
} } I make the coating around 0.003 to 0.006 mTorr.
} } Does this effect of spider web appear at this pressure?
} }
} }
} } Laci
} }
}
}



--
Jakab-Farkas Laszlo
Labortechnikus

SAPIENTIA - ERDÉLYI MAGYAR TUDOMÁNYEGYETEM
MÜSZAKI ÉS HUMÁNTUDOMÁNYOK KAR MAROSVÁSÁRHELY
Campus: Segesvári út 1C., Marosvásárhely/Koronka (a város határában)
Tel.: +40-(0)265-206210,+40-(0)265-208170, fax: 0265-206211
Mobil.: +40-(0)745-873844
E-mail: jflaci-at-ms.sapientia.ro
Postacím: 540485 Tîrgu Mures,op 9 cp4.


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From: jflaci-at-ms.sapientia.ro
Date: Sat, 16 Jan 2010 17:50:07 -0600
Subject: [Microscopy] Re: Choice for coating teeth structures for SEM ...

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

You ment this one?

http://www.amazon.com/Scanning-Electron-Microscopy-X-ray-Microanalysis/dp/0306472929

Laci

2010/1/17 Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} :
} Get a copy of Goldstein's book.
}
} JQuinn
}



--
Jakab-Farkas Laszlo
Labortechnikus

SAPIENTIA - ERDÉLYI MAGYAR TUDOMÁNYEGYETEM
MÜSZAKI ÉS HUMÁNTUDOMÁNYOK KAR MAROSVÁSÁRHELY
Campus: Segesvári út 1C., Marosvásárhely/Koronka (a város határában)
Tel.: +40-(0)265-206210,+40-(0)265-208170, fax: 0265-206211
Mobil.: +40-(0)745-873844
E-mail: jflaci-at-ms.sapientia.ro
Postacím: 540485 Tîrgu Mures,op 9 cp4.


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From: jflaci-at-ms.sapientia.ro
Date: Sat, 16 Jan 2010 18:04:14 -0600
Subject: [Microscopy] Re: Choice for coating teeth structures for SEM

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

Hello,

My coater is an old japanese glass bell jar system.
I usually work in high vacuum everywhere, including
the ion source, the vacuum test bench and the sputtering
system in the lab, microscopy is just a part of my activity.

The exact type of the coater I cannot recall at the moment I
am home now, not in the lab.

My problem is that i better would not heat up the tungsten
boat I use for evaporation in the presence of any oxigen,
and for the moment I do not have any inert gas in the lab,
as the gas containers are away at the vendor for safety routine
check. :-((.



2010/1/17 Gary Gaugler {gary-at-gaugler.com} :
} I did mean mT.  E-6 is territory I have not used.
} I just think that that is too high of vacuum.
} What do you get without the DP running?  Roughing
} pump only.
}
} What coater are you using?
}
} gary g.
}
}
} At 03:48 PM 1/16/2010, you wrote:
}
}
}
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} } To  Subscribe/Unsubscribe --
} } http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} }
} } ----------------------------------------------------------------------------
} }
} } I use an oil diffusion pump and a rotation pump.
} } I measure the pressure with an ionization gauge,
} } and I get a reading about 3-6 x 10 -6 Torr.
} }
} } By mT You ment mili Torr, or was I confused about that?
} }
} } Laci
} }
} } 2010/1/17 Gary Gaugler {gary-at-gaugler.com} :
} } } How can you get to 0.003-0.006mT?  If real, that
} } } is too high of vacuum.  For ultra fine coating, I
} } } work at 15mT with turbo pumped coater.
} } }
} } } gary g.
} } }
} } }
} } } At 03:26 PM 1/16/2010, you wrote:
} } }
} } }
} } }
} } } }
} } } }
} } } } ----------------------------------------------------------------------------
} } } } The Microscopy ListServer -- CoSponsor:  The Microscopy Society of
} } } } America
} } } } To  Subscribe/Unsubscribe --
} } } } http://www.microscopy.com/MicroscopyListserver
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} } } }
} } } }
} } } } ----------------------------------------------------------------------------
} } } }
} } } } Hello,
} } } }
} } } } I make the coating around 0.003 to 0.006 mTorr.
} } } } Does this effect of spider web appear at this pressure?
} } } }
} } } }
} } } } Laci
} } } }
} } }
} } }
} }
} }
} }
} } --
} } Jakab-Farkas Laszlo
} } Labortechnikus
} }
} } SAPIENTIA - ERDÉLYI MAGYAR TUDOMÁNYEGYETEM
} } MÜSZAKI ÉS HUMÁNTUDOMÁNYOK KAR MAROSVÁSÁRHELY
} } Campus: Segesvári út 1C., Marosvásárhely/Koronka (a város határában)
} } Tel.: +40-(0)265-206210,+40-(0)265-208170, fax: 0265-206211
} } Mobil.: +40-(0)745-873844
} } E-mail: jflaci-at-ms.sapientia.ro
} } Postacím: 540485 Tîrgu Mures,op 9 cp4.
} }
} }
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} } for SEM
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--
Jakab-Farkas Laszlo
Labortechnikus

SAPIENTIA - ERDÉLYI MAGYAR TUDOMÁNYEGYETEM
MÜSZAKI ÉS HUMÁNTUDOMÁNYOK KAR MAROSVÁSÁRHELY
Campus: Segesvári út 1C., Marosvásárhely/Koronka (a város határában)
Tel.: +40-(0)265-206210,+40-(0)265-208170, fax: 0265-206211
Mobil.: +40-(0)745-873844
E-mail: jflaci-at-ms.sapientia.ro
Postacím: 540485 Tîrgu Mures,op 9 cp4.


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From: kenconverse-at-qualityimages.biz
Date: Sun, 17 Jan 2010 07:23:37 -0600
Subject: [Microscopy] Re: Choice for coating teeth structures for SEM

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

Laci,
I'm a little confused. Are you evaporating or sputtering? If you're
evaporating, the vacuum you've stated is fine. The lower the pressure, the
better. If you're sputtering, it just won't work at all. Pressure in the
mT range is required.

As to coating thickness, the thicker it is, the more detail you'll hide. At
low mags 50nm might be OK, but for high mags I wouldn’t put on more than
5-10nm.

Any time there is a charging problem, you will want to reduce your beam size
even if you are working at low mags. On the JSM-5200 I normally do high mag
work with the probe size knob as close to fully CCW (about 7 o'clock) as I
can. If all is well with the system and the sample has decent signal, that
means fully CCW. Unless there is a need for x-ray generation or other
special needs (BSE), I tend to keep the probe size as small as possible for
all imaging (less than 12 o'clock), as this also tends to limit problems
with charging and/or beam damage to the specimen (not a significant problem
with teeth). Compensate for the noise (graininess) by using slower scan
speeds.

Also be sure that there is a conductive path from the top of the specimen to
the stub. If there is an area on the top of the specimen that is not of
interest, run some dag (graphite paint) from there, down the side of the
specimen, to the stub before coating. That way you can be sure that the
coating is connected to the stub. Without doing that, there is a chance
that the overhang of the specimen will cause a gap in the coating.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: jflaci-at-ms.sapientia.ro [mailto:jflaci-at-ms.sapientia.ro]
Sent: Saturday, January 16, 2010 7:06 PM
To: kenconverse-at-qualityimages.biz

Hello,

My coater is an old japanese glass bell jar system.
I usually work in high vacuum everywhere, including
the ion source, the vacuum test bench and the sputtering
system in the lab, microscopy is just a part of my activity.

The exact type of the coater I cannot recall at the moment I
am home now, not in the lab.

My problem is that i better would not heat up the tungsten
boat I use for evaporation in the presence of any oxigen,
and for the moment I do not have any inert gas in the lab,
as the gas containers are away at the vendor for safety routine
check. :-((.



2010/1/17 Gary Gaugler {gary-at-gaugler.com} :
} I did mean mT.  E-6 is territory I have not used.
} I just think that that is too high of vacuum.
} What do you get without the DP running?  Roughing
} pump only.
}
} What coater are you using?
}
} gary g.
}
}
} At 03:48 PM 1/16/2010, you wrote:
}
}
}
} }
} }
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} }
} } I use an oil diffusion pump and a rotation pump.
} } I measure the pressure with an ionization gauge,
} } and I get a reading about 3-6 x 10 -6 Torr.
} }
} } By mT You ment mili Torr, or was I confused about that?
} }
} } Laci
} }
} } 2010/1/17 Gary Gaugler {gary-at-gaugler.com} :
} } } How can you get to 0.003-0.006mT?  If real, that
} } } is too high of vacuum.  For ultra fine coating, I
} } } work at 15mT with turbo pumped coater.
} } }
} } } gary g.
} } }
} } }
} } } At 03:26 PM 1/16/2010, you wrote:
} } }
} } }
} } }
} } } }
} } } }
} } } }
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} } } }
} } } } Hello,
} } } }
} } } } I make the coating around 0.003 to 0.006 mTorr.
} } } } Does this effect of spider web appear at this pressure?
} } } }
} } } }
} } } } Laci
} } } }
} } }
} } }
} }
} }
} }
} } --
} } Jakab-Farkas Laszlo
} } Labortechnikus
} }
} } SAPIENTIA - ERDÉLYI MAGYAR TUDOMÁNYEGYETEM
} } MÜSZAKI ÉS HUMÁNTUDOMÁNYOK KAR MAROSVÁSÁRHELY
} } Campus: Segesvári út 1C., Marosvásárhely/Koronka (a város határában)
} } Tel.: +40-(0)265-206210,+40-(0)265-208170, fax: 0265-206211
} } Mobil.: +40-(0)745-873844
} } E-mail: jflaci-at-ms.sapientia.ro
} } Postacím: 540485 Tîrgu Mures,op 9 cp4.
} }
} }
} } ==============================Original
} } Headers==============================
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structures
} } for SEM
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--
Jakab-Farkas Laszlo
Labortechnikus

SAPIENTIA - ERDÉLYI MAGYAR TUDOMÁNYEGYETEM
MÜSZAKI ÉS HUMÁNTUDOMÁNYOK KAR MAROSVÁSÁRHELY
Campus: Segesvári út 1C., Marosvásárhely/Koronka (a város határában)
Tel.: +40-(0)265-206210,+40-(0)265-208170, fax: 0265-206211
Mobil.: +40-(0)745-873844
E-mail: jflaci-at-ms.sapientia.ro
Postacím: 540485 Tîrgu Mures,op 9 cp4.


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==============================Original Headers==============================
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From: jflaci-at-ms.sapientia.ro
Date: Sun, 17 Jan 2010 15:08:16 -0600
Subject: [Microscopy] Choice for coating teeth structures for SEM

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

Dear Ken!

Yes, I am working with evaporation. As I know, sputtering at this
vacuum levels (e-6) is almost impossible. I think I will try to make
some 5-10 nm
coatings, and see what is the result. Of course I have drawn two grounding lines
from conductive Ar paint to the base of the sample, for conducting
charge to the
stub.

Yeah, I have tried to go down to 7 o'clock with the beam size, but
at some point I couldn't obtain decent exposure. When the service
from Jeol was at our place, they said, that the scintillator is down to
almost 60%. Could this be the problem? At that point we did not have the
possibility to change it. :-(

Laci

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From: jflaci-at-ms.sapientia.ro
Date: Mon, 18 Jan 2010 01:51:46 -0600
Subject: [Microscopy] Choice for coating teeth structures for SEM

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

Laci,
It sounds like you have just about everything under control. Get a new
scintillator as soon as you can. Under most "normal" circumstances, an old
scint is not a big problem, but any time you are "pushing" things, it tends
to become far more important.

As is obvious, you can't use as small a spot size as you might like. A more
insidious effect is that the finest detail tends to come from the lowest
energy secondary electrons. They are the first ones to stop exciting the
scint as it ages, therefore your finest detail is the first to go.

Good luck!

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: jakabfarkaslaszlo-at-gmail.com [mailto:jakabfarkaslaszlo-at-gmail.com] On
Behalf Of Laszló Jakab-Farkas
Sent: Sunday, January 17, 2010 4:08 PM
To: Ken Converse
Cc: Microscopy-at-microscopy.com

And as tempting as it may seem to save money and install the scintillator
disk yourself, don't do it. It is expensive and easy to damage. It is
better to hire the JEOL tech to do it. If he (or she) screws it up, you are
not on the hook to buy a second scintillator disk. Don't ask me how I know
this 8-)

Ron L

-----Original Message-----
X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz]
Sent: Sunday, January 17, 2010 5:38 PM
To: lherault-at-bu.edu

Laci,
It sounds like you have just about everything under control. Get a new
scintillator as soon as you can. Under most "normal" circumstances, an old
scint is not a big problem, but any time you are "pushing" things, it tends
to become far more important.

As is obvious, you can't use as small a spot size as you might like. A more
insidious effect is that the finest detail tends to come from the lowest
energy secondary electrons. They are the first ones to stop exciting the
scint as it ages, therefore your finest detail is the first to go.

Good luck!

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: jakabfarkaslaszlo-at-gmail.com [mailto:jakabfarkaslaszlo-at-gmail.com] On
Behalf Of Laszló Jakab-Farkas
Sent: Sunday, January 17, 2010 4:08 PM
To: Ken Converse
Cc: Microscopy-at-microscopy.com

Hello everybody!

Thank You very much for the many answers,
and the help provided.

So in short the current status:

1. Yes, unfortunately the Ar paint was a typo, i ment Ag. Sorry about that.
2. I was worried about heating up the tungsten boat at 50mTorr
residual gas, not 50mTorr of Ar.
3. Thank You for noting that I shouldn't try to mount the scint by
myself, I think I feel what happened there, sorry to hear that.
4. Now I will try some Au coats armed with the information You kindly
provided, and

Many many thanks for everybody

Laci.

PS: both of the books are on the way to me, so I guess I will have to
read much in the short future.

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From: jacques.faerber-at-ipcms.u-strasbg.fr
Date: Mon, 18 Jan 2010 02:20:08 -0600
Subject: [Microscopy] Re: Choice for coating teeth structures for SEM

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

Hi

Two cents more.

If you coat your sample by evaporation and the sample is rough -typical
cases are a fracture of a porous rock, or the classical fly, your teeth
may be in that category-, it's important to be able to rock the sampe
during the coating. Evaporation from a boat is a point source, as
sputtering is a diffuse/large surface source. With evaporation you may
have much shadowing, and a non continuous film on the sample, what can
explain the remaining charging. Increasing the thickness will not solve
that problem. As Ken said, 5-10 nm should be enough, a bit more for low
mag.
You need to rock the sample, in a way that the most of the sides of
holes, facet, etc will be exposed to the metal flux. If the rocking is
not possible, you can do 2 or 3 short evaporations, at different
incidences, for a total thickness of ay 20 nm, but it will take a longer
time.
These are two reason wy sputtering is most prefered to evaporation,
directionnality and time spending.

Hope it helps

J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr



jflaci-at-ms.sapientia.ro a écrit :
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear Ken!
}
} Yes, I am working with evaporation. As I know, sputtering at this
} vacuum levels (e-6) is almost impossible. I think I will try to make
} some 5-10 nm
} coatings, and see what is the result. Of course I have drawn two grounding lines
} from conductive Ar paint to the base of the sample, for conducting
} charge to the
} stub.
}
} Yeah, I have tried to go down to 7 o'clock with the beam size, but
} at some point I couldn't obtain decent exposure. When the service
} from Jeol was at our place, they said, that the scintillator is down to
} almost 60%. Could this be the problem? At that point we did not have the
} possibility to change it. :-(
}
} Laci
}
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}

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From: donald.gibbon-at-matcoinc.com
Date: Mon, 18 Jan 2010 12:24:24 -0600
Subject: [Microscopy] Fwd: FW: Ask-A-Microscopist

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

Hi David

Thanks for your updated email and I do hope ALL SEM people read it as you
have hit the nail on the head (as we would say in England).

I am sure that you and thousands of others around the world do everything
that you can to ensure your colleagues know as much as possible about the
instruments you use. But, from the off line questions that I have, not one
has mentioned a level of understanding in this complex area that I would
have hoped had been achieved. Which brings us to your most important point,
there are people around who will be able to answer most questions; in
particular the equipment manufacturer!

Back to cars so that I do not offend anyone. How many of you feel that you
are a competent driver, perhaps on a race track you could get round pretty
quickly? I am sure that I or one of our race team could knock at least 14%
off your lap time; relate that to how much extra I am sure that you could
learn from your SEM manufacturer!

I have thought about this over the weekend and although I earn my living by
passing on my knowledge I am setting out below an example that may help
people to better understand the double detection system of a modern SEM.

The customer of a client had nanotubes that were uncoated and they were
required to be imaged at 150,000X. The client had worked at 2kV, 4mm WD,
60um aperture and the upper detector, in a Zeiss microscope that uses a
range of aperture sizes to control probe current. The specimen material was
cast upon a double sided carbon tape. My client's problem was that all the
micrographs were demonstrating a considerable amount of charge which often
overwhelmed the image detail.

My thoughts as we tackled the problem are written in brackets with the
action undertaken being numbered.

(Starting with the specimen preparation, I do not like double sided carbon
take above 100,000X as I feel the current density becomes too high and
introduces material instability. The main problem is that using the desired
150,000X probably requires the upper detector and unfortunately upper
detectors are able to ignore SE3. SE3 are important because they are
converted backscattered electron information and backscattered electrons, to
a certain extent, ignore surface charge. The conversion comes about through
the BSE making contact with the lens and other chamber components producing
secondary electrons to a level directly related to the amount of backscatter
at any one image point. Had the magnification requirement been below
80,000X we could have used the lower detector because it receives a higher
level of SE3 thus charge free operation may have been possible. People in
my experience do not use the lower detector enough, they leap in possessed
with the quest for high resolution, when the lower detector may give them
the information with less problems. However we needed a starting point which
was the clients settings with a few subtle changes).

(We needed to work under as many anti-charge parameters as possible.
Staying away from a bundle of fibres and only using those that were in
intimate contact with the substrate was my first change. Fibre bundles are
a nightmare as you have very poor contact with earth. Then I instructed
that we should work on the left hand side of a bundle to minimise discharge
from two problem areas (i) The scan on most machines spends more time on the
left hand side of the screen so always work to leave your last used area to
the right. (ii) The last area you looked at would also be charged. Try
leaving your visited areas with problems on the right it does actually
minimise discharge. But first of all we needed to lower the specimen/WD.)


1. Work at 10mm, chamber/lower detector, select the edge of the bundle
of nanotubes and work with bundles and previously used areas to the right of
the screen.

(Results - poor resolution but no charge, so try to use a smaller
aperture/probe current if there is insufficient resolution, as expected,
then we will needed to move to the upper detector.)

2. Set up with the 30um aperture and lower detector.

(Improved but not good enough.)

3. Switch between lower and upper detector and compare results.

(Upper detector had insufficient signal therefore shortening the WD should
improve the situation.)

4. Move to 9mm WD

(Better performance, signal is still a little poor, we need a bigger probe
current, but still no charge, naturally we moved to an new area for each
part of this experiment.)

5. Using the 60um aperture the image is quite good, but not good
enough.

(We need Reduce the WD again)

6. Reduced to 8mm WD and the image is much stronger with little or no
charge.

(Resolution is pretty good at a fast scan at 150,000X with a low level of
charge at image recording speeds. Reducing to the smaller aperture may help
but I was against moving to a shorter working distance at this stage.)

7. Using the 30um aperture the resolution was improved and we were able
to use the appropriate image recording speeds without charge, the image was
a little too noisy for me.

(We had been using 2kV so now was the time to reduce the noise level through
experimenting with the accelerating voltage. Remember moving from 2 to 3 kV
is the equivalent to moving from 10 to 15kV in different circumstances?)

8. We stepped from 2kV to 2.1kV, to 2.2kV, to 2.3kV and then at 2.4kV
but here the charge level started to interfere with the image during image
recording.

(So we now had the kV correct at 2.3 and the probe current but what about
the WD, could we improve the performance further without charge?)

9. We moved to 7mm WD but even trying different areas of the specimen
we were not satisfied with the amount of charge in the recorded images.

The results were presented to the client's customer who had, it appears, run
the specimen with another laboratory using exactly the same instrument. He
was very surprised at the quality of result my client produced, claiming he
could see more detail than he had initially thought possible.

So there you are just a few hints and tips that those who asked have gained.

To summarise STATE OF THE ART SEM I hope you all agree with David and I that
it is better to ASK and you should receive, be it a consultant or your
instrument manufacturer.

Happy scanning

Steve


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com






-----Original Message-----
X-from: M. David Frey [mailto:freym2-at-rpi.edu]
Sent: 15 January 2010 19:53
To: protrain-at-emcourses.com
Cc: Microscopy-at-microscopy.com

Hi Fred

I agree with all that you mention and I think it is important as a vendor to
work hard on judging your customer.

If I run a basic residential course very few wish to attend, but run an
advanced course and that creates interest. However when I start this course
from the same data that is i the basic course the "advanced" users are
fascinated because they too did not know! What I think we may be saying is
that not all customers are up to speed on SEM in general and therefore it is
tough for them to understand a State of the Art instrument.

But back to my first email. When you sell a State of the Art piece of
equipment you owe it to your client to bring them up to State of the Art or
in my mind what is the point of selling the equipment? All of us who have
been in the business know that the more you sell, the more you sell. With
that aim the better trained your new customer is the more papers they will
write and the good name of your product is enhanced. That was the
philosophy with which I had always worked and operating as I do now, often
through personal recommendation, I agree it is important to "do a good job".


Lets await more comment

Steve



-----Original Message-----
X-from: freym2-at-rpi.edu [mailto:freym2-at-rpi.edu]
Sent: 14 January 2010 19:33
To: protrain-at-emcourses.com

Debby,

Thanks for adding to what I have written. I often find here at RPI, also an
academic environment that the skills and knowledge a user brings into the
training that is provided helps them go much farther in the quality of the
images they produce. It also helps if they are interested in learning more
than steps 1 through 20 in how to get an image. Understanding the "basics"
of electron microscopy is a must.

X-from my prior experience as an applications person for a microscope
company,
we often made many assumptions to the user's basic knowledge of electron
microscopy. As an aps person, we didn't always have time to fully educate
the customer on the "basics" of electron microscopy, we had 2 or 3 days to
get them to "know" their microscope, and then we off to home and our
families or on to the next customer.

I do like the fact that there is some requirement at Purdue for the users to
have a full credit course and lab behind them before they have full access
to the systems. Here at RPI there are a few people trying to push that sort
of requirement forward. I just always remind each user as they get trained
that they can always call, asking for help is good, and no question is a
stupid question.

One last idea I'd like to offer is that I take their science away from them
when teaching users to use the microscope. I make them look at my samples,
and they are focused on learning the basics, the controls, and the
microscope itself. They aren't asking why didn't my experiment work, they
are asking how do I make the picture better.

I offer that we should take the following conclusion away from this
discussion: Microscope vendors train users to user their products. They
aren't always to best source of basic electron microscopy education. (I
think the Norm Burns said this often at Cambridge/LEICA/LEO/Zeiss). As a
vendor we should leave it to the likes of Steve, Lehigh, or the old PASEM
course, or the New Paltz course at the Mountain House, or the classes
offered at our respective/favorite academic intuitions of choice. The
microscope vendors have a wonderful pool of knowledgeable people who are
tasked with selling and supporting a very technical product. At the end of
the day, month, or year the bottom line to microscope vendors is how many
did we sell, and how many happy customers do we have.*

David

* You can look at this as if it is driver's education, you don't see Ford,
or VW teaching driver's education, they sell cars. When you pick up your new
car they'll show you the basics, where the key goes, and such but it is
assumed you know the rules of the road and have driver's license. :)

M. David Frey
Senior Application Engineer
Rensselaer Polytechnic Institute
Low Center for Industrial Inn.
Center for Integrated Electronics
110 8th Street
CII 4161 (Office)
CII 6015 (Packages and Mail)
Troy, NY 12180
518-276-3323 (office)
518-698-2288 (mobile)

-----Original Message-----
X-from: Sherman, Debra M [mailto:dsherman-at-purdue.edu]
Sent: Thursday, January 14, 2010 1:25 PM
To: freym2-at-rpi.edu

Dear Eric: This is a subject that has been of great interest to me over
the years. I suggest the way to approach this is by making replicas of
the snow crystals. Collodion in Amyl Acetate will do the trick very
nicely. Put your glass slides and a dropper-bottle of say 2% collodion
in amyl acetate in a box that you can put outside to drop to ambient
temperature. When it snows, coat a slide with a couple of drops of the
solution and hold it out in the snow to catch flakes. When you have a
few, put the slide back in the box, still at ambient temperature but
protected from more snow. The amyl acetate will vaporize, the collodion
will harden and the snow will eventually sublime, leaving behind a
perfect replica of the snow flakes, excellent for light microscopy...
and even SEM, should you want to go to higher magnification. This takes
some fiddling around to perfect, but I guarantee it works very well. I
used it to make my Christmas cards once long ago.

Donald L. Gibbon

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Friday, January 15, 2010 4:57 PM
To: Gibbon, Donald L.

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of
MSA, and any reply should go directly to the poster as well as to the
list.
==========================================================

} Subject: FW: Ask-A-Microscopist
} Date: Fri, 15 Jan 2010 16:04:41 -0500
}
} -----Original Message-----
} From: Eric Coates [mailto:ericcoates-at-hotmail.com]
} Sent: Tuesday, December 01, 2009 1:35 PM
} To: AssociationManagement
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Tuesday, December 01,
} 2009 at 10:35:14 AM.
}
} realname - Eric Coates
} Email - ericcoates-at-hotmail.com
} SUBJECT_OF_QUESTION - freeze substitution
} QUESTION - I am looking for a way to perform freeze substitution of ice
} crystals for optical, not SEM, analysis. I am familiar with one method,
} described in the article "Low Temperature SEM of Precipitated and
} Metamorphosed Snow Crystals Collected and Transported from Remote
Sites"
} that appeared in JAMA v2 n3 1996, but it involves osmium tetroxide,
} which I would like to avoid. I have also tried silver nitrate with no
} success. Any suggestions would be appreciated.
}
} Thanks
}
} Eric Coates
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: s.h.coetzee-at-gmail.com
Date: Tue, 19 Jan 2010 00:51:31 -0600
Subject: [Microscopy] Choice for coating teeth structures for SEM

Contents Retrieved from Microscopy Listserver Archives
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I was involved in quite a lot of teeth work some long time ago. Teeth
are porous and result in two things, charging and SLOW pump down.
These trapped gasses also contaminated the surface during imaging.
Those days we did not had the fancy lower dirty vacuum options like
today. We resulted in leaving the sample in a high vacuum coater
overnight to get a really good pump down, then C-coated it, flushed
the chamber with Ar gas or dry nitrogen gas till room pressure after
coating. Au-Pd coated it afterwards over the C at the highest
possible vacuum the coater will work. Then 10kv at the lowest
possible probe current we could get a reasonable image.

Today I would go for C-coat and low kv feg in a low vacuum mode if I
have the option. But I will not bypass the C-coat prior to another
coating technique since you get a fairly even film at great vacuum
levels.

Just a early morning thought.

Stephan Coetzee
EM Scientist
EMU
University of Botswana


On Sun, Jan 17, 2010 at 1:19 AM, {gary-at-gaugler.com} wrote:
}
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} Au is not my choice for anything being coated.  It produces
} a spider web characteristic if not done at high vacuum.
} Try Au/Pd or Ir at 30mT and don't worry all that much
} about film thickness.  50nm or thereabouts should work fine.
}
} gary g.
}
}
} At 02:05 PM 1/16/2010, you wrote:
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--
Stephan H Coetzee
Chief Technician
Electron Microscope Unit
University of Botswana


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From: oshel1pe-at-cmich.edu
Date: Tue, 19 Jan 2010 08:47:39 -0600
Subject: [Microscopy] Inverted LM recommendation wanted

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Listers,

Since I'm doing "Ask a Microscopist" now, I don't feel I should give
recommendations. Perhaps some of the folks Down Under would care to
respond?

Phil

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of
MSA, and any reply should go directly to the poster as well as to the
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==========================================================
} -----Original Message-----
} From: L Wu [mailto:wehi618-at-yahoo.com]
} Sent: Tuesday, January 19, 2010 3:21 AM
} To: AssociationManagement
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Tuesday, January 19, 2010
} at 12:20:31 AM.
}
} realname - L Wu
} Email - wehi618-at-yahoo.com
} ORGANIZATION - University of Melbourne
} EDUCATION - Graduate College
} LOCATION - Melbourne, Australia
} SUBJECT_OF_QUESTION - Inverted microscope
} QUESTION - The lab I am working in is planning to purchase an Inverted
} Flourescent Microscope. The models we are interested are Nikon Ti-S and
} Olympus IX71. We'd appreciate information or comments on the Pro and
} Conts of these two models.
}
} Many thanks.
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: jeff-at-tss-consulting.com
Date: Tue, 19 Jan 2010 09:41:55 -0600
Subject: [Microscopy] PhD,TEM Scientist, looking for work in Massachusetts

Contents Retrieved from Microscopy Listserver Archives
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I represent an excellent scientist with PhD and 6 years experience in
private and public companies, post degree. Analytic skills on TEM plus
failure analysis and quality control experience, X-Ray Diffraction etc.
Employers please contact me directly at jeff-at-tss-consulting.com or
941-475-6270.


Jeff West

Managing Partner/East Coast

TSS Consulting Ltd.

(year round ) 877-489-2425

(summer office) 207-925-6014

(winter office) 941-475-6270

(c) 781-956-9913

www.tss-consulting.com {http://www.tss-consulting.com/}

jeff-at-tss-consulting.com



"SEMPER PARATUS" (ALWAYS PREPARED)





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From: svmcphie-at-utmb.edu
Date: Tue, 19 Jan 2010 16:12:45 -0600
Subject: [Microscopy] Research associate position

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

A Research Associate position is available immediately in the laboratory of Svetla Stoilova-McPhie to study the structure and conformations of membrane-bound blood coagulation complexes by combining Cryo-electron microscopy and biophysical methods.
The main project will involve 2D and helical crystallization of blood coagulation factors and complexes onto functionalized lipid monolayers and lipid nanotubes, followed by Cryo-EM and structure analysis. The lipid-protein systems will be also characterised by various light scattering and spectroscopic techniques to elucidate fully the structure-function relationship in the context of membrane-binding. The Sealy Centre for Structural Biology and Molecular Biophysics at UTMB http://www.scsb.utmb.edu/, includes also a state-of-the art Cryo-EM facility. This project is furthermore in collaboration with the NCMI centre at Baylor College of Medicine, equipped with the latest technology in Cryo-EM and structure analysis: http://ncmi.bcm.tmc.edu/ncmi/. The lab has the latest model CD-spectrophotometer with fluorescence and Peltier device (Jasco J815), as well as all protein and tissue culture facilities; fluorescence and confocal microscopy.
The ideal candidate should have strong background in protein biochemistry and interest in structural biology. The successful applicant should have previous experience in protein biochemistry and analytical methods for protein analysis. Proficiency in basic biochemical techniques, HPLC, SDS-PAGE and UV-VIS spectroscopy are necessary. Excellent time management skills, strong communication and interpersonal skills are essential. Prior experience in lipid chemistry and membrane biology, as well as in fluorescence, CD and confocal microscopy or/and structural biology will be a plus, but not essential.
Candidates with Ph.D. or M.Sc. considering furthering their research career at the interface between biomedical sciences and nanotechnology are encouraged to apply.
Please send your current CV and three references to Dr. Svetla Stoilova-McPhie at svmcphie-at-utmb.edu {mailto:svmcphie-at-utmb.edu} , Department of Neuroscience and Cell Biology, University of Texas Medical Branch at Galveston: www.utmb.edu {http://www.utmb.edu/}
UTMB is also part of the Gulf Coast Consortia http://cohesion.rice.edu/centersandinst/gcc/

Svetla Stoilova-McPhie, PhD
Assistant Professor,
Department of Neuroscience and Cell Biology
Scientist, Sealy Centre for Structural Biology
and Molecular Biophysics
University of Texas Medical Branch at Galveston
301 University Boulevard, Galveston, Texas 77555-0620
Cell: (+1) 281-229-2261
Fax: (1+) 409-747-2200
Email: svmcphie-at-utmb.edu


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From: Stacy.Gates-at-utoledo.edu
Date: Tue, 19 Jan 2010 20:05:07 -0600
Subject: [Microscopy] viaWWW: SEM GIVEAWAY!

Contents Retrieved from Microscopy Listserver Archives
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Email: Stacy.Gates-at-utoledo.edu
Name: Stacy

Organization: University of Toledo Instrumentation Center

Title-Subject: [Filtered] SEM GIVEAWAY!

Message: The University of Toledo is looking to pass on our current
SEM to another institution. The instrument has been fully functional
and under service contract until January 2010; when it was shut down
in preparation for the new instrument. The instrument we are looking
to pass on is a JEOL JSM- 6100. We are not charging a fee for the
instrument itself, however it will be the responsibility of the
recipient to fund any transport fees associated with the move. If you
are interested, or know someone who may be interested, in acquiring
this piece of equipment please contact the University of Toledo
Instrumentation Center at (419) 530-7899 or (419) 530-7487
immediately; as this will be handled on a first come first serve basis

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From: drg.mitchell-at-sydney.edu.au
Date: Sat, 23 Jan 2010 08:59:21 -0600
Subject: [Microscopy] viaWWW: TEM: EELSTools plugin for DigitalMicrograph

Contents Retrieved from Microscopy Listserver Archives
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Email: drg.mitchell-at-sydney.edu.au
Name: David Mitchell

Organization: EMU, University of Sydney

Title-Subject: [Filtered] TEM: EELSTools plugin for DigitalMicrograph

Message: Dear All
If you use Gatan's DigitalMicrograph for EELS spectroscopy, then the
EELSTools plugin for DigitalMicrograph may be of interest to you. It
was jointly developed over several years by myself and Bernhard
Schaffer. The software consists of a choice of plugins for
hardware-connected or offline systems; an example EELS data cube to
experiment with; a detailed instruction manual and an uninstaller.
Installation is straightforward and no scripting knowledge is needed.
The package includes twenty different tools for acquiring EELS data
from Gatan Energy Filters, processing/measuring/formatting spectra
and a number of useful calculators for calculating and displaying
EELS data. The software can be downloaded free of charge from the
following URL:
http://www.dmscripting.com/eelstools.html
Regards
Dave Mitchell

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From: msanders-at-umn.edu
Date: Mon, 25 Jan 2010 14:58:29 -0600
Subject: [Microscopy] viaWWW: Full-time technical position Univ. of Minn.

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Email: msanders-at-umn.edu
Name: Mark Sanders

Organization: University of Minnesota, Imaging Center

Title-Subject: [Filtered] Full-time technical position

Message: The Imaging Center in the College of Biological Sciences at
University of Minnesota (http://www.cbs.umn.edu/ic/) is hiring a
full-time technical staff member who will work the many users of this
established facility.

We're looking for someone with a solid understanding of the
principles of microscopy and experience in modern forms of
microscopy--in particular scanning electron microscopy and/or
confocal and widefield microscopy, image analysis, processing and
output. The ideal candidate would have a strong background in both
microscopy, biology and sample preparation, with an enthusiasm for
computing, cutting-edge knowledge of microscopy and imaging and good
people/communications skills.

Those interested should apply on-line at
http://www1.umn.edu/ohr/employment . The job requisition number is:
164820 .

The University of Minnesota is an equal opportunity educator and employer.

Feel free to contact me off list (msanders-at-umn.edu) with any questions.

Thanks!

Mark Sanders
msanders-at-umn.edu
Program Director
CBS Imaging Center

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From: d33-at-ornl.gov
Date: Mon, 25 Jan 2010 16:11:27 -0600
Subject: [Microscopy] viaWWW: SEM Outreach Opportunity Needed: NYC/metro area

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Email: d33-at-ornl.gov
Name: Donovan N. Leonard

Organization: UT-Knoxville/ORNL Mat. Sci.

Title-Subject: [Filtered] SEM Outreach Opportunity Needed: NYC/metro area

Message: Dear Colleagues,

The Pratt Institute (Brooklyn, NY) architecture
research studio is requesting a few hours of SEM
time in the NYC metro area for 10 students. The
professor for the course, Jonas Coersmeier, has
created a novel interdisciplinary design and
research studio in which students utilize micro
and nano-scale patterns found in nature as
inspiration for long span architectural
structures. If interested, involvement in studio
discourse as guest critic or lecturer is
possible. In the past students have chosen their
own samples and acquired micrographs themselves
with a SEM, then used features from the
micrographs as base units for larger
architectural design. The research studio
students have blogged about their experiences
learning about nanotechnology, electron
microscopy and the process of architectural
design. Please visit any of the blogs listed
below or reference the Microscopy Today article
to learn more about this novel design studio.

If you can donate some time for students on a SEM
to acquire micrographs for their design studio
project, please contact Prof. Jonas Coersmeier by
email at ëjpc61-at-columbia.eduí. This is a great
outreach opportunity.

Many thanks in advance.
Donovan

ps -I am posting this for Jonas because in the
past I have been involved with his design studio
and found it an enriching experience. Anytime
students, not involved STEM related studies, can
be inspired and excited by microscopy is, it is
worth the extra effort. Iím hoping to continue my
collaboration with Pratt and would be happy to
provide more information or answer questions if
you contact me offline at ëd33-at-ornl.goví.

Microscopy Today Article
http://www.probelog.com/span/MT-Coersmeier_Leonard.pdf
Design Studio Blogs
http://www.probelog.com/span/
http://www.probelog.com/sem/


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From: wei.kong-at-oregonstate.edu
Date: Mon, 25 Jan 2010 17:31:34 -0600
Subject: [Microscopy] viaWWW: Open postdoctoral position - Oregon State Un.

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Email: wei.kong-at-oregonstate.edu
Name: Wei Kong

Organization: Oregon State University

Title-Subject: [Filtered] Open postdoctoral position

Message: An immediate opening for a postdoctoral position is
available at Oregon State University. The project involves electron
diffraction of laser oriented molecules embedded in superfluid helium
droplets. Experience in coherent x-ray or electron diffraction is
preferred but not mandated. The individual needs to have a solid
background in physics, optics, and electronics, and extensive hands
on experience in one of the fields. Intellectual curiosity and
willingness to step into unfamiliar disciplines are essential
qualities. Candidates should send their curriculum vitae with 3
reference letters to Wei Kong, Department of Chemistry, Oregon State
University, Corvallis, OR 97331. The position is open until filled.
Oregon State University is an AA/EOE.

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From: r-holdford-at-ti.com
Date: Mon, 25 Jan 2010 18:16:07 -0600
Subject: [Microscopy] Texas Society for Microscopy Spring Meeting Apr. 15-17, 2010

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Announcing the 2010 Meeting of THE TEXAS SOCIETY FOR MICROSCOPY
“Embracing All Forms of Microscopy”

Meeting Dates: April 15-17, 2010
Hilton Garden Inn, Frisco, TX
First Call for Papers

Invited Speakers:
Freshman Phage Hunters: Integrating a Research-based Experience into
Freshman Biology for Majors by Dr. Lee Hughes

Failure Analysis Strategies in Electronics Industry - Past, Present and
Future by Dr. Puligandla Viswanadham

Other information and registration forms can be found on our web site:
http://www.texasmicroscopy.org/.

--
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Becky Holdford (r-holdford-at-ti.com)
972-995-2360
972-648-8743 (pager)
SC Packaging FA
Texas Instruments, Inc.
13536 N. Central Expressway MS 940
Dallas, TX 75243
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~


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From: John.Mardinly-at-wdc.com
Date: Mon, 25 Jan 2010 18:54:10 -0600
Subject: [Microscopy] Liquid Nitrogen Safety

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Well, it has been a long time since I have seen anything about Liquid
Nitrogen Safety on the listserver, but when I stumbled across this, I just
had to share it. Apparently at SLAC, which is the Stanford Linear
Accelerator, they have a SLAC Family Day, SLAC Take our children to Work
Day, during which they make ice cream by mixing cream, sugar and LIQUID
NITROGEN in bowls for kids, with children right there. They even let the
children STICK THEIR FINGERS IN THE LIQUID NITROGEN (well, only for one
second)! I'm not making this up! Here is a web site describing it:
http://keithjobe.com/ice-cream.html

Well, I would not let MY daughter stick her finger in liquid nitrogen, but
apparently some parents are, although the Stanford safety people seem to
take a dim view of this.


John Mardinly
Western Digital



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From: frah0010-at-umn.edu
Date: Mon, 25 Jan 2010 19:20:07 -0600
Subject: [Microscopy] Re: Liquid Nitrogen Safety

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Has everyone seen the videos of individuals who put liquid nitrogen in their mouths and then spit it out?

http://www.metacafe.com/watch/1847229/spitting_liquid_nitrogen/
http://www.youtube.com/watch?v=rnFu8OSWk-A
http://www.youtube.com/watch?v=i2xYB9xPCtc
http://www.metacafe.com/watch/1290462/liquid_nitrogen/

The idea is that one's mouth or hand is so hot compared to the liquid nitrogen that there is a thin gaseous layer of nitrogen isolating the skin from the liquid nitrogen.

But I sure wouldn't do it, and I don't encourage anyone else to try it either.

Best,
Ellery

-----------------
Ellery Frahm
Senior Research Fellow, Department of Geology & Geophysics
Manager & Principal Analyst, Electron Microprobe Lab
University of Minnesota - Twin Cities campus
Lab website: http://probelab.geo.umn.edu/
Personal: http://web.mac.com/elleryfrahm/



On Jan 25, 2010, at 6:58 PM, John.Mardinly-at-wdc.com wrote:

}
}
}
} ----------------------------------------------------------------------------
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} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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}
} Well, it has been a long time since I have seen anything about Liquid
} Nitrogen Safety on the listserver, but when I stumbled across this, I just
} had to share it. Apparently at SLAC, which is the Stanford Linear
} Accelerator, they have a SLAC Family Day, SLAC Take our children to Work
} Day, during which they make ice cream by mixing cream, sugar and LIQUID
} NITROGEN in bowls for kids, with children right there. They even let the
} children STICK THEIR FINGERS IN THE LIQUID NITROGEN (well, only for one
} second)! I'm not making this up! Here is a web site describing it:
} http://keithjobe.com/ice-cream.html
}
} Well, I would not let MY daughter stick her finger in liquid nitrogen, but
} apparently some parents are, although the Stanford safety people seem to
} take a dim view of this.
}
}
} John Mardinly
} Western Digital
}
}
}
} ==============================Original Headers==============================
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From: bozzola-at-siu.edu
Date: Mon, 25 Jan 2010 19:30:01 -0600
Subject: [Microscopy] Re: Liquid Nitrogen Safety

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John,

OMG, the liquid nitrogen ice cream URL looks like a lawsuit waiting to
happen. Liquid nitrogen and chillin' are a dangerous combination.
(Sorry, but the pun just jumped out!)


Some may have heard of Dippin' Dots, a commercially-available ice
cream produced by flash freezing droplets of ice cream using liquid
nitrogen. The inventor, Curt D. Jones, is a microbiologist and
graduate of our university. I have not yet tried the ice cream but
hear that it's delightful.

John Bozzola
Chillin' at The SIU IMAGE Center
Carbondale, IL

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From: wesaia-at-iastate.edu
Date: Mon, 25 Jan 2010 20:21:29 -0600
Subject: [Microscopy] Re: Liquid Nitrogen Safety

Contents Retrieved from Microscopy Listserver Archives
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Some chemical engineering students here at Iowa State University developed a process for using LN2 to make really creamy ice cream back in 1999. They were probably fooling around in the lab for a while before that. One of the principals, Will Schroeder, used our JEOL 840A SEM under my tutelage. Here's their web site. http://www.blueskycreamery.com/ or you can check out a NY Times article on them at http://www.nytimes.com/2005/08/10/dining/10nitr.html

I've poured LN2 across my hands. I haven't tried sticking my finger into a dewar of it - yet. I know that the extreme temperature difference is what keeps you from freezing. I had a prof in a mass transfer class explaining it as the leidenfrost effect (http://en.wikipedia.org/wiki/Leidenfrost_effect). It's what causes water droplets to dance around on a very hot griddle whereas they would quickly spread and evaporate on a not so hot griddle. The vapor layor that forms really slows the heat transfer. Even then, the droplet does eventually heat up and steam away - and so would your finger chill and freeze.

Like they say, don't try this at home, but I am sure some will anyway.

Warren Straszheim
________________________________________
X-from: bozzola-at-siu.edu [bozzola-at-siu.edu]
Sent: Monday, January 25, 2010 7:30 PM
To: wesaia-at-iastate.edu

John,

OMG, the liquid nitrogen ice cream URL looks like a lawsuit waiting to
happen. Liquid nitrogen and chillin' are a dangerous combination.
(Sorry, but the pun just jumped out!)


Some may have heard of Dippin' Dots, a commercially-available ice
cream produced by flash freezing droplets of ice cream using liquid
nitrogen. The inventor, Curt D. Jones, is a microbiologist and
graduate of our university. I have not yet tried the ice cream but
hear that it's delightful.

John Bozzola
Chillin' at The SIU IMAGE Center
Carbondale, IL

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From: richard.ross-at-allisontransmission.com
Date: Tue, 26 Jan 2010 07:53:10 -0600
Subject: [Microscopy] Re: Liquid Nitrogen Safety

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

Way back when, Chuck Fiori used to put on a demo with LN2 at the LeHigh
SEM school. Chuck wanted to show that sometimes its more hazardous to trap
LN2 against the body with protective gear than to give the liedenfrost
effect room to create its protective blanket of gas. If the LN2 was
provided, he would eventually take some into his mouth and blow 'smoke'
rings. He told of a time when he accidentally swallowed some of the LN2
and the resulting stomach gas pressure caused him to black out, falling to
the stage unconscious. Ultimately the excess pressure relieved itself as a
belch and Chuck came to. Chuck certainly taught me things in ways that I
remember!

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From: W.Muss-at-salk.at
Date: Tue, 26 Jan 2010 11:14:23 -0600
Subject: [Microscopy] Re: Liquid Nitrogen Safety

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

Good morning, good afternoon, good evening - hopefully any somewhere will apply,
hello, dear colleagues,

Just to add my 2 €-cents (apologize if the message became too long):

Yes, we can discuss about children STICK THEIR FINGERS IN THE LIQUID NITROGEN and other stuff.... in general,
since in EM we sometimes have to work with 'cruel' things and substances (and others a little bit more often).
Cryogenics also CAN be hazardous (as } hot { water is/can be), depending on how we are } working with { it.
If we couldn't do that safe(ly) it necessarily / perhaps would have been forbidden for a long time.

There are a lot of sources on the web one can find concerning use and misuse of e. g. liquid nitrogen:
for example:

http://www.altair.org/hazard.html (DONT DIE ! Laboratory Hazards: Safety, Prevention, First Aid, C 2001-2010)

http://www.reachoutmichigan.org/funexperiments/agesubject/lessons/nitrogen.html
( "Fun experiments" website last updated 2003)

http://smb.slac.stanford.edu/users_guide/manual/Experiment_policies.html#SECTION00027000000000000000
(describes hazards and proper handling procedures for work with liquid nitrogen, 2010)

http://stores.biochem.uiowa.edu/Pages/ln2msds.htm
("old" but IMO "good" example for a US-Univ's Safety Data Sheet)

http://www.chaosscience.org.uk/dem/public_html//article.php?story=20031216175107931
(Liquid Nitrogen Risk Assessment, 16/12/03 )

Also, some Labs are questioning in advance for safety issues on the things someone will bring into the Lab:
e.g. http://www.sbc.anl.gov/pdfs/hazard_assessment_form.pdf (take a look for 'cryogens', Argonne Natl. Lab, SBC Hazard Assessment Form), perhaps just to be on the safe side for their own staff people and Lab's interior.

AND finally, YES, LN2-ingestion - to take a mouthfull of LN2 and blast it off - only for the EXPERIENCED CAN BE a junky-funky trick, BUT -
on the other side - for the unexperienced (unfortunately also for the 'careless' experienced I have to admit!) - it could be not only dangerous but LIFE threatening: cf.
http://pediatrics.aappublications.org/cgi/reprint/105/1/121
( Benjamin Z. Koplewitz, et al.... Gastric Perforation Attributable to Liquid Nitrogen Ingestion,
in: Pediatrics 2000;105;121-123, open Access)

A similar situation has been reported on http://www.darwinawards.com/personal/personal2000-25.html,
which eventually was } awarded { with the DARWIN AWARD in 2000 (==} cit: "The Darwin Awards salute the improvement of the human genome by honoring those who accidentally remove themselves from it..." end of cit.)

I personally have no doubt that it IS possible (and I have done that for demonstration of "Leidenfrost phenomenon" a lot of times way back...) to place one or two fingers, eventually(and forsure) the whole hand into liquid nitrogen for 1-2 seconds, PROVIDED some necessary PREREQUISITES which - IN ADVANCE - have to be theoretically explained and are to be met to/for those } trying { such an "adventure" and IMO only should be done in a SUPERVISED situation.

Despite (or better: BECAUSE) respecting all those "little" {problem-makers} in our daily work
(if this is not done by other staff members, I have to to do such work literally by myself) I am not in a blue funk of perhaps desastrous results of using/handling all those substances like
OsO4 (hazard, toxic), p-phenylenediamine (pesticide, toxic), uranyl-acetate from stock powder (radioactive), cryogens (like LN2, precooled isobutane, also Freon gas [some residual inventory] if the pressurized can will be used inverted) and other substances/chemicals (mostly used in very small quantities), not to forget all those "may be hazardous resins" out in the dark...

WHY I am NOT AFRAID/do not fear those: since I have learned - before handling and using those -
WHAT the respective properties of materials/substances/fluids are,
HOW they have ( practically ) to be used safely,
WHAT the consequences eventually will be (personal and for others) if handled unsafe, and
HOW to dispose of (remnants, by-products of reactions, etc.) properly.

(as an example for awareness: read: http://www.hull.ac.uk/chemstores/coshhadv.html (COSHH- Control of Substances Hazardous to Health, Univ.Hull, UK: 'part of the U.K.'s CRIMINAL LAW' !)

Anybody who has {studied} properties of chemicals etc., their handling and use properly and correctly will know how to proceed with the use of material and substances we use in an EM-lab. If this has not be done, no one should do } things { he never got explained and does not know about consequences in case of misuse.

There IMO is no need to disallow or eventually prohibit (necessary) actions which to the unexperienced (perhaps) will be harmful, but to the experienced does not apply seriously.
So, last but not least: it is the responsibilty of the "experienced" to teach the unexperienced... so you as the {experienced} decide what can be done/shown and what CANNOT be done /shown.

Concerning: Ice-Cream-experiments with LN2, those perhaps are an "interesting" demonstration for a new practical application but easily can/could be safely substituted by {old fashioned} ice-cream-making (e.g lowering the temperature of the fluids by using eg. the pot-freezer method: the temperature of the ingredients is reduced by placing them inside a tub filled with ice and salt) Cf. http://en.wikipedia.org/wiki/Ice_cream .

At the end: just only pointing to the long MSA thread on } N2 gas-LN2 { starting May 2nd 2009
Conc: [Microscopy] Nitrogen leak (JEOL 6701F SEM)


Regards,
Wolfgang MUSS
Head EM-Lab, Pathology
SALK-LKH (Gen. Hosp.)
SALZBURG Austria




} -----Ursprüngliche Nachricht-----
} Von: richard.ross-at-allisontransmission.com
} [mailto:richard.ross-at-allisontransmission.com]
} Gesendet: Dienstag, 26. Jänner 2010 14:57
} An: Muß Wolfgang
} Betreff: [Microscopy] Re: Liquid Nitrogen Safety [Ice-Cream made with
} LN2, concerns about]
}
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} Way back when, Chuck Fiori used to put on a demo with LN2 at the LeHigh SEM school.
Chuck wanted to show that sometimes its more hazardous to trap LN2 against the body with protective gear than to give the liedenfrost effect room to create its protective blanket of gas. If the LN2 was provided, he would eventually take some into his mouth and blow 'smoke' rings. He told of a time when he accidentally swallowed some of the LN2 and the resulting stomach gas pressure caused him to black out, falling to the stage unconscious.
Ultimately the excess pressure relieved itself as a belch and Chuck came to.
Chuck certainly taught me things in ways that I remember! {
}
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From: oshel1pe-at-cmich.edu
Date: Tue, 26 Jan 2010 11:54:48 -0600
Subject: [Microscopy] Re: Liquid Nitrogen Safety

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

LN2 safety has a long history in the microscopy list archives. As
Wolfgang points out, learn to handle LN2 (or whatever) safely, and
don't worry about it. The Leidenfrost effect and LN2 is well known
and well discussed (here and elsewhere). And yes, I demonstrate this
myself in our SEM labs. It's a non-issue. (Just make sure your hand
is dry -- unlike when you stick it into molten lead to demonstrate
the Leidenfrost effect.)
Getting stuck in an elevator with a 160L LN2 transport dewar is much
more of a potential problem.

As for LN2 and ice cream, I'm amused that people would get worried
about safety issues. There are commercial ice cream parlors and
restaurants making LN2 ice cream. Part of the molecular-gastronomy
movement.
http://www.urbandaddy.com/chi/1533/iCream_Chicago_CHI_Stone_Cold_Creamery_UrbanDaddy_Archives
http://blogs.villagevoice.com/forkintheroad/archives/2009/10/lulu_mookys_bri.php
http://www.starchefs.com/features/liquid-nitrogen/html/index.shtml
etcetera
LN2 ice cream is made tableside in some places. I don't think we
really need to worry about it.

If you do want a hazard to worry about, think of
ethane/butane/propane as LN2 cooled cryogens and how they *don't*
exhibit the Leidenfrost effect, and how they do enrich themselves in
oxygen by condensing it out of the air. And the potential of
oxygen-enriched ethane ...

Phil
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: sawyert-at-science.oregonstate.edu
Date: Tue, 26 Jan 2010 14:48:06 -0600
Subject: [Microscopy] electronic calendar

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

We are interested in setting up an electron scheduling calendar for our
EM lab. Currently we have four microscopes and some sample preparation
tools users need to sign-up to use. FOM (fomnetworks.com) has been
suggested but I am wondering about other programs that might be cheaper
or free and still offer similar benefits: scheduling, tracking beam time
etc.

Please offer any suggestions off line

Thanks
Teresa







--
Teresa Sawyer
Electron Microscope Facility Manager
Oregon State University
541-737-5245
sawyert-at-science.oregonstate.edu
Cordely Hall 1078
http://www.science.oregonstate.edu/bpp/EMfacility/index.htm


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From: dcromey-at-email.arizona.edu
Date: Tue, 26 Jan 2010 15:10:30 -0600
Subject: [Microscopy] electronic calendar

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

Teresa,

Someone asked me this sort of question earlier this month. We use the OCF
software here, since it was written at our University.

The computing facility for AZ Research Labs (provided core facilities to
campus) developed the OCF scheduler software. While it doesn't do
everything, it does have the virtue of being freely available as open source
software. Obviously there will be a financial investment in IT
time/resources for anyone who wants to use OCF, but we like it here. For
more information, see: http://demo.arl.arizona.edu/

Other labs I know of use either iCal or CALcium from Brownbear Software. I
use iCal for low-traffic instruments, but I don't have any experience with
CALcium. See: http://www.brownbearsw.com/

I've heard of other software scheduling/billing/management software, but I
have no experience with it:

PPMS (Pasteur/Rockefeller Platform Management System) - http://ppms.info/

FACES - http://faces.ccrc.uga.edu/

Some places use Google, or Yahoo calendars, or if they have a Microsoft
Exchange server, calendars in Outlook.

Good luck,
Doug

^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Douglas W. Cromey, M.S. - Assistant Scientific Investigator
Dept. of Cell Biology & Anatomy, University of Arizona
1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA

office: AHSC 4212 email: Cromey-at-Arizona.edu
voice: 520-626-2824 fax: 520-626-2097

http://swehsc.pharmacy.arizona.edu/exppath/
Home of: "Microscopy and Imaging Resources on the WWW"


-----Original Message-----
X-from: sawyert-at-science.oregonstate.edu
[mailto:sawyert-at-science.oregonstate.edu]
Sent: Tuesday, January 26, 2010 1:49 PM
To: dcromey-at-email.arizona.edu

We are interested in setting up an electron scheduling calendar for our
EM lab. Currently we have four microscopes and some sample preparation
tools users need to sign-up to use. FOM (fomnetworks.com) has been
suggested but I am wondering about other programs that might be cheaper
or free and still offer similar benefits: scheduling, tracking beam time
etc.

Please offer any suggestions off line

Thanks
Teresa







--
Teresa Sawyer
Electron Microscope Facility Manager
Oregon State University
541-737-5245
sawyert-at-science.oregonstate.edu
Cordely Hall 1078
http://www.science.oregonstate.edu/bpp/EMfacility/index.htm


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From: kenconverse-at-qualityimages.biz
Date: Tue, 26 Jan 2010 15:47:45 -0600
Subject: [Microscopy] Re: Liquid Nitrogen Safety

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

Phil,
Thank you. I've enjoyed playing with lN2 from time to time. I also got 2nd
degree burns from it once when I was not playing with it. The problem was
that I acted before I thought. I realized my mistake milliseconds after my
inappropriate action and milliseconds before acquiring my burns.

The trick, of course, is to know what you are about and to THINK (in time).

I also like to occasionally play with He or Ar when it's available. The
voice changes are fun, but I've read some real kill-joy things about those
activities, also. Again, if you THINK and realize that you're displacing
O2, then you breathe a number of times between inhaling either gas. We can
still demonstrate scientific principles, we can still have fun and we can
still inspire kids to opt for a career where you get to play with cool toys
and other playthings.

The GPS display stuck on a windshield is probably more hazardous than the
above activities.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Tuesday, January 26, 2010 12:56 PM
To: kenconverse-at-qualityimages.biz

LN2 safety has a long history in the microscopy list archives. As
Wolfgang points out, learn to handle LN2 (or whatever) safely, and
don't worry about it. The Leidenfrost effect and LN2 is well known
and well discussed (here and elsewhere). And yes, I demonstrate this
myself in our SEM labs. It's a non-issue. (Just make sure your hand
is dry -- unlike when you stick it into molten lead to demonstrate
the Leidenfrost effect.)
Getting stuck in an elevator with a 160L LN2 transport dewar is much
more of a potential problem.

As for LN2 and ice cream, I'm amused that people would get worried
about safety issues. There are commercial ice cream parlors and
restaurants making LN2 ice cream. Part of the molecular-gastronomy
movement.
http://www.urbandaddy.com/chi/1533/iCream_Chicago_CHI_Stone_Cold_Creamery_Ur
banDaddy_Archives
http://blogs.villagevoice.com/forkintheroad/archives/2009/10/lulu_mookys_bri
.php
http://www.starchefs.com/features/liquid-nitrogen/html/index.shtml
etcetera
LN2 ice cream is made tableside in some places. I don't think we
really need to worry about it.

If you do want a hazard to worry about, think of
ethane/butane/propane as LN2 cooled cryogens and how they *don't*
exhibit the Leidenfrost effect, and how they do enrich themselves in
oxygen by condensing it out of the air. And the potential of
oxygen-enriched ethane ...

Phil
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: donald.gibbon-at-matcoinc.com
Date: Tue, 26 Jan 2010 16:31:13 -0600
Subject: [Microscopy] Re: Liquid Nitrogen Safety

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




As a semi-professional ice cream maker, I'd like to second Wolfgang Muss's reference to the old-fashioned but utterly reliable and safe method of using a hand-cranked ice cream freezer rather than any other option. There are reasons why this works well, which, if ignored, explain why it doesn't work sometimes! Making ice cream is an exercise in heat transfer and crystal growth. The heat transfer surface is the metal wall of the tub which is rotated by the crank. Counter-rotating inside the tube is a pair of wooden paddles, keeping that surface clean and free of ice, constantly removing the freezing cream. The idea is to get a eutectic mixture of salt and ice on the outside, i.e., minimum possible temperature in this system, dump in the cream and start cranking. Don't ever stop for even one second. i.e., keep that surface clean, and in six-eight minutes your job will be done. The idea is to maximize the number of crystals and minimize their size. It is NOT a tedious process, hardly enough to get winded.

My most sought-after publication in my entire life was my paper on The Thermodynamics of Homemade Ice Cream, published in the Journal Of Chemical Education. I still have the thermocouple-fitted ice cream freezer in which we made many gallons of ice cream, doing the experimental work for this paper. Somebody had to do it! And sticking your finger in the ice cream will not harm you at all!

Donald L. Gibbon

-----Original Message-----
X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at]
Sent: Tuesday, January 26, 2010 12:23 PM
To: Gibbon, Donald L.

Good morning, good afternoon, good evening - hopefully any somewhere will apply,
hello, dear colleagues,

Just to add my 2 €-cents (apologize if the message became too long):

Yes, we can discuss about children STICK THEIR FINGERS IN THE LIQUID NITROGEN and other stuff.... in general,
since in EM we sometimes have to work with 'cruel' things and substances (and others a little bit more often).
Cryogenics also CAN be hazardous (as } hot { water is/can be), depending on how we are } working with { it.
If we couldn't do that safe(ly) it necessarily / perhaps would have been forbidden for a long time.

There are a lot of sources on the web one can find concerning use and misuse of e. g. liquid nitrogen:
for example:

http://www.altair.org/hazard.html (DONT DIE ! Laboratory Hazards: Safety, Prevention, First Aid, C 2001-2010)

http://www.reachoutmichigan.org/funexperiments/agesubject/lessons/nitrogen.html
( "Fun experiments" website last updated 2003)

http://smb.slac.stanford.edu/users_guide/manual/Experiment_policies.html#SECTION00027000000000000000
(describes hazards and proper handling procedures for work with liquid nitrogen, 2010)

http://stores.biochem.uiowa.edu/Pages/ln2msds.htm
("old" but IMO "good" example for a US-Univ's Safety Data Sheet)

http://www.chaosscience.org.uk/dem/public_html//article.php?story=20031216175107931
(Liquid Nitrogen Risk Assessment, 16/12/03 )

Also, some Labs are questioning in advance for safety issues on the things someone will bring into the Lab:
e.g. http://www.sbc.anl.gov/pdfs/hazard_assessment_form.pdf (take a look for 'cryogens', Argonne Natl. Lab, SBC Hazard Assessment Form), perhaps just to be on the safe side for their own staff people and Lab's interior.

AND finally, YES, LN2-ingestion - to take a mouthfull of LN2 and blast it off - only for the EXPERIENCED CAN BE a junky-funky trick, BUT -
on the other side - for the unexperienced (unfortunately also for the 'careless' experienced I have to admit!) - it could be not only dangerous but LIFE threatening: cf.
http://pediatrics.aappublications.org/cgi/reprint/105/1/121
( Benjamin Z. Koplewitz, et al.... Gastric Perforation Attributable to Liquid Nitrogen Ingestion,
in: Pediatrics 2000;105;121-123, open Access)

A similar situation has been reported on http://www.darwinawards.com/personal/personal2000-25.html,
which eventually was } awarded { with the DARWIN AWARD in 2000 (==} cit: "The Darwin Awards salute the improvement of the human genome by honoring those who accidentally remove themselves from it..." end of cit.)

I personally have no doubt that it IS possible (and I have done that for demonstration of "Leidenfrost phenomenon" a lot of times way back...) to place one or two fingers, eventually(and forsure) the whole hand into liquid nitrogen for 1-2 seconds, PROVIDED some necessary PREREQUISITES which - IN ADVANCE - have to be theoretically explained and are to be met to/for those } trying { such an "adventure" and IMO only should be done in a SUPERVISED situation.

Despite (or better: BECAUSE) respecting all those "little" {problem-makers} in our daily work
(if this is not done by other staff members, I have to to do such work literally by myself) I am not in a blue funk of perhaps desastrous results of using/handling all those substances like
OsO4 (hazard, toxic), p-phenylenediamine (pesticide, toxic), uranyl-acetate from stock powder (radioactive), cryogens (like LN2, precooled isobutane, also Freon gas [some residual inventory] if the pressurized can will be used inverted) and other substances/chemicals (mostly used in very small quantities), not to forget all those "may be hazardous resins" out in the dark...

WHY I am NOT AFRAID/do not fear those: since I have learned - before handling and using those -
WHAT the respective properties of materials/substances/fluids are,
HOW they have ( practically ) to be used safely,
WHAT the consequences eventually will be (personal and for others) if handled unsafe, and
HOW to dispose of (remnants, by-products of reactions, etc.) properly.

(as an example for awareness: read: http://www.hull.ac.uk/chemstores/coshhadv.html (COSHH- Control of Substances Hazardous to Health, Univ.Hull, UK: 'part of the U.K.'s CRIMINAL LAW' !)

Anybody who has {studied} properties of chemicals etc., their handling and use properly and correctly will know how to proceed with the use of material and substances we use in an EM-lab. If this has not be done, no one should do } things { he never got explained and does not know about consequences in case of misuse.

There IMO is no need to disallow or eventually prohibit (necessary) actions which to the unexperienced (perhaps) will be harmful, but to the experienced does not apply seriously.
So, last but not least: it is the responsibilty of the "experienced" to teach the unexperienced... so you as the {experienced} decide what can be done/shown and what CANNOT be done /shown.

Concerning: Ice-Cream-experiments with LN2, those perhaps are an "interesting" demonstration for a new practical application but easily can/could be safely substituted by {old fashioned} ice-cream-making (e.g lowering the temperature of the fluids by using eg. the pot-freezer method: the temperature of the ingredients is reduced by placing them inside a tub filled with ice and salt) Cf. http://en.wikipedia.org/wiki/Ice_cream .

At the end: just only pointing to the long MSA thread on } N2 gas-LN2 { starting May 2nd 2009
Conc: [Microscopy] Nitrogen leak (JEOL 6701F SEM)


Regards,
Wolfgang MUSS
Head EM-Lab, Pathology
SALK-LKH (Gen. Hosp.)
SALZBURG Austria




} -----Ursprüngliche Nachricht-----
} Von: richard.ross-at-allisontransmission.com
} [mailto:richard.ross-at-allisontransmission.com]
} Gesendet: Dienstag, 26. Jänner 2010 14:57
} An: Muß Wolfgang
} Betreff: [Microscopy] Re: Liquid Nitrogen Safety [Ice-Cream made with
} LN2, concerns about]
}
} --------------------------------------------------------------------------
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} Way back when, Chuck Fiori used to put on a demo with LN2 at the LeHigh SEM school.
Chuck wanted to show that sometimes its more hazardous to trap LN2 against the body with protective gear than to give the liedenfrost effect room to create its protective blanket of gas. If the LN2 was provided, he would eventually take some into his mouth and blow 'smoke' rings. He told of a time when he accidentally swallowed some of the LN2 and the resulting stomach gas pressure caused him to black out, falling to the stage unconscious.
Ultimately the excess pressure relieved itself as a belch and Chuck came to.
Chuck certainly taught me things in ways that I remember! {
}
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From: kenconverse-at-qualityimages.biz
Date: Tue, 26 Jan 2010 17:14:59 -0600
Subject: [Microscopy] Re: Liquid Nitrogen Safety

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

Don,
You're right! It's a dirty job, but somebody's got to do it. My wife and I got an electric ice cream maker as a wedding gift that we haven't used nearly enough. My favorite recipe not only includes the heavy cream, but a half dozen eggs. To die for...

Of course the kill-joys will tell us that the heavy cream, the cholesterol in the eggs and all the sugar will kill us, not to mention the environmental effects of all the cattle and chickens, plus the inhumane conditions they are kept in.

I'll have another serving of that custard ice cream recipe, thank you.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: donald.gibbon-at-matcoinc.com [mailto:donald.gibbon-at-matcoinc.com]
Sent: Tuesday, January 26, 2010 5:34 PM
To: kenconverse-at-qualityimages.biz




As a semi-professional ice cream maker, I'd like to second Wolfgang Muss's reference to the old-fashioned but utterly reliable and safe method of using a hand-cranked ice cream freezer rather than any other option. There are reasons why this works well, which, if ignored, explain why it doesn't work sometimes! Making ice cream is an exercise in heat transfer and crystal growth. The heat transfer surface is the metal wall of the tub which is rotated by the crank. Counter-rotating inside the tube is a pair of wooden paddles, keeping that surface clean and free of ice, constantly removing the freezing cream. The idea is to get a eutectic mixture of salt and ice on the outside, i.e., minimum possible temperature in this system, dump in the cream and start cranking. Don't ever stop for even one second. i.e., keep that surface clean, and in six-eight minutes your job will be done. The idea is to maximize the number of crystals and minimize their size. It is NOT a tedious process, !
hardly enough to get winded.

My most sought-after publication in my entire life was my paper on The Thermodynamics of Homemade Ice Cream, published in the Journal Of Chemical Education. I still have the thermocouple-fitted ice cream freezer in which we made many gallons of ice cream, doing the experimental work for this paper. Somebody had to do it! And sticking your finger in the ice cream will not harm you at all!

Donald L. Gibbon

-----Original Message-----
X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at]
Sent: Tuesday, January 26, 2010 12:23 PM
To: Gibbon, Donald L.

Good morning, good afternoon, good evening - hopefully any somewhere will apply,
hello, dear colleagues,

Just to add my 2 €-cents (apologize if the message became too long):

Yes, we can discuss about children STICK THEIR FINGERS IN THE LIQUID NITROGEN and other stuff.... in general,
since in EM we sometimes have to work with 'cruel' things and substances (and others a little bit more often).
Cryogenics also CAN be hazardous (as } hot { water is/can be), depending on how we are } working with { it.
If we couldn't do that safe(ly) it necessarily / perhaps would have been forbidden for a long time.

There are a lot of sources on the web one can find concerning use and misuse of e. g. liquid nitrogen:
for example:

http://www.altair.org/hazard.html (DONT DIE ! Laboratory Hazards: Safety, Prevention, First Aid, C 2001-2010)

http://www.reachoutmichigan.org/funexperiments/agesubject/lessons/nitrogen.html
( "Fun experiments" website last updated 2003)

http://smb.slac.stanford.edu/users_guide/manual/Experiment_policies.html#SECTION00027000000000000000
(describes hazards and proper handling procedures for work with liquid nitrogen, 2010)

http://stores.biochem.uiowa.edu/Pages/ln2msds.htm
("old" but IMO "good" example for a US-Univ's Safety Data Sheet)

http://www.chaosscience.org.uk/dem/public_html//article.php?story=20031216175107931
(Liquid Nitrogen Risk Assessment, 16/12/03 )

Also, some Labs are questioning in advance for safety issues on the things someone will bring into the Lab:
e.g. http://www.sbc.anl.gov/pdfs/hazard_assessment_form.pdf (take a look for 'cryogens', Argonne Natl. Lab, SBC Hazard Assessment Form), perhaps just to be on the safe side for their own staff people and Lab's interior.

AND finally, YES, LN2-ingestion - to take a mouthfull of LN2 and blast it off - only for the EXPERIENCED CAN BE a junky-funky trick, BUT -
on the other side - for the unexperienced (unfortunately also for the 'careless' experienced I have to admit!) - it could be not only dangerous but LIFE threatening: cf.
http://pediatrics.aappublications.org/cgi/reprint/105/1/121
( Benjamin Z. Koplewitz, et al.... Gastric Perforation Attributable to Liquid Nitrogen Ingestion,
in: Pediatrics 2000;105;121-123, open Access)

A similar situation has been reported on http://www.darwinawards.com/personal/personal2000-25.html,
which eventually was } awarded { with the DARWIN AWARD in 2000 (==} cit: "The Darwin Awards salute the improvement of the human genome by honoring those who accidentally remove themselves from it..." end of cit.)

I personally have no doubt that it IS possible (and I have done that for demonstration of "Leidenfrost phenomenon" a lot of times way back...) to place one or two fingers, eventually(and forsure) the whole hand into liquid nitrogen for 1-2 seconds, PROVIDED some necessary PREREQUISITES which - IN ADVANCE - have to be theoretically explained and are to be met to/for those } trying { such an "adventure" and IMO only should be done in a SUPERVISED situation.

Despite (or better: BECAUSE) respecting all those "little" {problem-makers} in our daily work
(if this is not done by other staff members, I have to to do such work literally by myself) I am not in a blue funk of perhaps desastrous results of using/handling all those substances like
OsO4 (hazard, toxic), p-phenylenediamine (pesticide, toxic), uranyl-acetate from stock powder (radioactive), cryogens (like LN2, precooled isobutane, also Freon gas [some residual inventory] if the pressurized can will be used inverted) and other substances/chemicals (mostly used in very small quantities), not to forget all those "may be hazardous resins" out in the dark...

WHY I am NOT AFRAID/do not fear those: since I have learned - before handling and using those -
WHAT the respective properties of materials/substances/fluids are,
HOW they have ( practically ) to be used safely,
WHAT the consequences eventually will be (personal and for others) if handled unsafe, and
HOW to dispose of (remnants, by-products of reactions, etc.) properly.

(as an example for awareness: read: http://www.hull.ac.uk/chemstores/coshhadv.html (COSHH- Control of Substances Hazardous to Health, Univ.Hull, UK: 'part of the U.K.'s CRIMINAL LAW' !)

Anybody who has {studied} properties of chemicals etc., their handling and use properly and correctly will know how to proceed with the use of material and substances we use in an EM-lab. If this has not be done, no one should do } things { he never got explained and does not know about consequences in case of misuse.

There IMO is no need to disallow or eventually prohibit (necessary) actions which to the unexperienced (perhaps) will be harmful, but to the experienced does not apply seriously.
So, last but not least: it is the responsibilty of the "experienced" to teach the unexperienced... so you as the {experienced} decide what can be done/shown and what CANNOT be done /shown.

Concerning: Ice-Cream-experiments with LN2, those perhaps are an "interesting" demonstration for a new practical application but easily can/could be safely substituted by {old fashioned} ice-cream-making (e.g lowering the temperature of the fluids by using eg. the pot-freezer method: the temperature of the ingredients is reduced by placing them inside a tub filled with ice and salt) Cf. http://en.wikipedia.org/wiki/Ice_cream .

At the end: just only pointing to the long MSA thread on } N2 gas-LN2 { starting May 2nd 2009
Conc: [Microscopy] Nitrogen leak (JEOL 6701F SEM)


Regards,
Wolfgang MUSS
Head EM-Lab, Pathology
SALK-LKH (Gen. Hosp.)
SALZBURG Austria




} -----Ursprüngliche Nachricht-----
} Von: richard.ross-at-allisontransmission.com
} [mailto:richard.ross-at-allisontransmission.com]
} Gesendet: Dienstag, 26. Jänner 2010 14:57
} An: Muß Wolfgang
} Betreff: [Microscopy] Re: Liquid Nitrogen Safety [Ice-Cream made with
} LN2, concerns about]
}
} --------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} Way back when, Chuck Fiori used to put on a demo with LN2 at the LeHigh SEM school.
Chuck wanted to show that sometimes its more hazardous to trap LN2 against the body with protective gear than to give the liedenfrost effect room to create its protective blanket of gas. If the LN2 was provided, he would eventually take some into his mouth and blow 'smoke' rings. He told of a time when he accidentally swallowed some of the LN2 and the resulting stomach gas pressure caused him to black out, falling to the stage unconscious.
Ultimately the excess pressure relieved itself as a belch and Chuck came to.
Chuck certainly taught me things in ways that I remember! {
}
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From: Robert.Brandom-at-Bruker-AXS.com
Date: Tue, 26 Jan 2010 19:41:11 -0600
Subject: [Microscopy] viaWWW: NESM -- February 9 Dinner Meeting

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

I prefer the hand-cranked version, myself. Not only does it give more
direct tactile feedback on the overall thickness of the ice cream, but it
gives the baby (To me) cousins something to do that is out of the way of my
kitchen chaos on Thanksgiving!

As for the health aspect, there was a study that I read within the last
three or four years (I can't remember anything specific) that said that the
largest contributor to weight gain when eating is the mental state of the
eater. I was discussing this with a colleague of mine and we came to the
conclusion that it must be true, based on the French. (My apologies at this
generalization, but I am basing my experience on the four incredible weeks I
spent wandering around the French countryside...)

The French eat wonderfully decadent, flavorful, and rich foods, and drink
gallons of truly elegant wine. Neither me nor my colleague remembered
seeing any obese French people (We're sure they exist, but they don't seem
to be the norm as it seems in the US). We also experienced the wonderful
French culture of taking your time while eating, enjoying both the meal and
the company, not rushing through it.

We could also come up with more French nationals who lived to respectable
old age than we could of any other nationality.

I know that this is in no way a scientific observation, but it's still
enough to make me want to wander around rural France for the rest of my life
eating hand-churned ice cream...

Great. Now I'm hungry. First time this list has done that to me...

--Justin A. Kraft
Professional Lab Rat/High Energy Physics Server Monkey
Florida International University
Miami, FL


--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar

n Tue, Jan 26, 2010 at 6:19 PM, |--kenconverse-at-qualityimages.biz--| wrote:

--|
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--|
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---
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--|
--| Don,
--| You're right! It's a dirty job, but somebody's got to do it. My wife an=
d
--| I got an electric ice cream maker as a wedding gift that we haven't used
--| nearly enough. My favorite recipe not only includes the heavy cream, but=
a
--| half dozen eggs. To die for...
--|
--| Of course the kill-joys will tell us that the heavy cream, the cholestero=
l
--| in the eggs and all the sugar will kill us, not to mention the environmen=
tal
--| effects of all the cattle and chickens, plus the inhumane conditions they
--| are kept in.
--|
--| I'll have another serving of that custard ice cream recipe, thank you.
--|
--| Ken Converse
--| owner
--|
--| QUALITY IMAGES
--| Servicing Scanning Electron Microscopes
--| Since 1981
--| 474 So. Bridgton Rd.
--| Bridgton, ME 04009
--| 207-647-4348
--| Fax 207-647-2688
--| kenconverse-at-qualityimages.biz
--| qualityimages.biz
--|
--|
--| -----Original Message-----
--| X-from: donald.gibbon-at-matcoinc.com [mailto:donald.gibbon-at-matcoinc.com]
--| Sent: Tuesday, January 26, 2010 5:34 PM
--| To: kenconverse-at-qualityimages.biz
--| Subject: [Microscopy] FW: Re: Liquid Nitrogen Safety
--|
--|
--|
--|
--|
--| -------------------------------------------------------------------------=
---
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a
--| To Subscribe/Unsubscribe --
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--|
--|
--|
--|
--| As a semi-professional ice cream maker, I'd like to second Wolfgang Muss'=
s
--| reference to the old-fashioned but utterly reliable and safe method of us=
ing
--| a hand-cranked ice cream freezer rather than any other option. There are
--| reasons why this works well, which, if ignored, explain why it doesn't wo=
rk
--| sometimes! Making ice cream is an exercise in heat transfer and crystal
--| growth. The heat transfer surface is the metal wall of the tub which is
--| rotated by the crank. Counter-rotating inside the tube is a pair of woode=
n
--| paddles, keeping that surface clean and free of ice, constantly removing =
the
--| freezing cream. The idea is to get a eutectic mixture of salt and ice on =
the
--| outside, i.e., minimum possible temperature in this system, dump in the
--| cream and start cranking. Don't ever stop for even one second. i.e., keep
--| that surface clean, and in six-eight minutes your job will be done. The i=
dea
--| is to maximize the number of crystals and minimize their size. It is NOT =
a
--| tedious process, !
--| hardly enough to get winded.
--|
--| My most sought-after publication in my entire life was my paper on The
--| Thermodynamics of Homemade Ice Cream, published in the Journal Of Chemica=
l
--| Education. I still have the thermocouple-fitted ice cream freezer in whic=
h
--| we made many gallons of ice cream, doing the experimental work for this
--| paper. Somebody had to do it! And sticking your finger in the ice cream w=
ill
--| not harm you at all!
--|
--| Donald L. Gibbon
--|
--| -----Original Message-----
--| X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at]
--| Sent: Tuesday, January 26, 2010 12:23 PM
--| To: Gibbon, Donald L.
--| Subject: [Microscopy] Re: Liquid Nitrogen Safety
--|
--|
--|
--|
--|
--| -------------------------------------------------------------------------=
---
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a
--| To Subscribe/Unsubscribe --
--| http://www.microscopy.com/MicroscopyListserver
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--|
--| -------------------------------------------------------------------------=
---
--|
--| Good morning, good afternoon, good evening - hopefully any somewhere will
--| apply,
--| hello, dear colleagues,
--|
--| Just to add my 2 =C3=83=C2=A2=C3=A2=E2=82=AC=C5=A1=C3=82=C2=AC-cents (apo=
logize if the message became too long):
--|
--| Yes, we can discuss about children STICK THEIR FINGERS IN THE LIQUID
--| NITROGEN and other stuff.... in general,
--| since in EM we sometimes have to work with 'cruel' things and substances
--| (and others a little bit more often).
--| Cryogenics also CAN be hazardous (as --|hot|-- water is/can be),
depending on
--| how we are --|working with|-- it.
--| If we couldn't do that safe(ly) it necessarily / perhaps would have been
--| forbidden for a long time.
--|
--| There are a lot of sources on the web one can find concerning use and
--| misuse of e. g. liquid nitrogen:
--| for example:
--|
--| http://www.altair.org/hazard.html (DONT DIE ! Laboratory Hazards: Safety=
,
--| Prevention, First Aid, C 2001-2010)
--|
--|
--| http://www.reachoutmichigan.org/funexperiments/agesubject/lessons/nitroge=
n.html
--| ( "Fun experiments" website last updated 2003)
--|
--|
--| http://smb.slac.stanford.edu/users_guide/manual/Experiment_policies.html#=
SECTION00027000000000000000
--| (describes hazards and proper handling procedures for work with liquid
--| nitrogen, 2010)
--|
--| http://stores.biochem.uiowa.edu/Pages/ln2msds.htm
--| ("old" but IMO "good" example for a US-Univ's Safety Data Sheet)
--|
--|
--| http://www.chaosscience.org.uk/dem/public_html//article.php?story=3D20031=
216175107931
--| (Liquid Nitrogen Risk Assessment, 16/12/03 )
--|
--| Also, some Labs are questioning in advance for safety issues on the thing=
s
--| someone will bring into the Lab:
--| e.g. http://www.sbc.anl.gov/pdfs/hazard_assessment_form.pdf (take a look
--| for 'cryogens', Argonne Natl. Lab, SBC Hazard Assessment Form), perhaps j=
ust
--| to be on the safe side for their own staff people and Lab's interior.
--|
--| AND finally, YES, LN2-ingestion - to take a mouthfull of LN2 and blast it
--| off - only for the EXPERIENCED CAN BE a junky-funky trick, BUT -
--| on the other side - for the unexperienced (unfortunately also for the
--| 'careless' experienced I have to admit!) - it could be not only dangerous
--| but LIFE threatening: cf.
--| http://pediatrics.aappublications.org/cgi/reprint/105/1/121
--| ( Benjamin Z. Koplewitz, et al.... Gastric Perforation Attributable to
--| Liquid Nitrogen Ingestion,
--| in: Pediatrics 2000;105;121-123, open Access)
--|
--| A similar situation has been reported on
--| http://www.darwinawards.com/personal/personal2000-25.html,
--| which eventually was --|awarded|-- with the DARWIN AWARD in 2000
(=3D=3D--|cit:=
"The
--| Darwin Awards salute the improvement of the human genome by honoring thos=
e
--| who accidentally remove themselves from it..." end of cit.)
--|
--| I personally have no doubt that it IS possible (and I have done that for
--| demonstration of "Leidenfrost phenomenon" a lot of times way back...) to
--| place one or two fingers, eventually(and forsure) the whole hand into liq=
uid
--| nitrogen for 1-2 seconds, PROVIDED some necessary PREREQUISITES which - I=
N
--| ADVANCE - have to be theoretically explained and are to be met to/for tho=
se
--| --|trying|-- such an "adventure" and IMO only should be done in a SUPERVISED
--| situation.
--|
--| Despite (or better: BECAUSE) respecting all those "little" |--problem-maker=
s--|
--| in our daily work
--| (if this is not done by other staff members, I have to to do such work
--| literally by myself) I am not in a blue funk of perhaps desastrous result=
s
--| of using/handling all those substances like
--| OsO4 (hazard, toxic), p-phenylenediamine (pesticide, toxic), uranyl-aceta=
te
--| from stock powder (radioactive), cryogens (like LN2, precooled isobutane,
--| also Freon gas [some residual inventory] if the pressurized can will be u=
sed
--| inverted) and other substances/chemicals (mostly used in very small
--| quantities), not to forget all those "may be hazardous resins" out in the
--| dark...
--|
--| WHY I am NOT AFRAID/do not fear those: since I have learned - before
--| handling and using those -
--| WHAT the respective properties of materials/substances/fluids are,
--| HOW they have ( practically ) to be used safely,
--| WHAT the consequences eventually will be (personal and for others) if
--| handled unsafe, and
--| HOW to dispose of (remnants, by-products of reactions, etc.) properly.
--|
--| (as an example for awareness: read:
--| http://www.hull.ac.uk/chemstores/coshhadv.html (COSHH- Control of
--| Substances Hazardous to Health, Univ.Hull, UK: 'part of the U.K.'s CRIMI=
NAL
--| LAW' !)
--|
--| Anybody who has |--studied--| properties of chemicals etc., their
handling an=
d
--| use properly and correctly will know how to proceed with the use of mater=
ial
--| and substances we use in an EM-lab. If this has not be done, no one shoul=
d
--| do --|things|-- he never got explained and does not know about
consequences i=
n
--| case of misuse.
--|
--| There IMO is no need to disallow or eventually prohibit (necessary) actio=
ns
--| which to the unexperienced (perhaps) will be harmful, but to the experien=
ced
--| does not apply seriously.
--| So, last but not least: it is the responsibilty of the "experienced" to
--| teach the unexperienced... so you as the |--experienced--| decide
what can be
--| done/shown and what CANNOT be done /shown.
--|
--| Concerning: Ice-Cream-experiments with LN2, those perhaps are an
--| "interesting" demonstration for a new practical application but easily
--| can/could be safely substituted by |--old fashioned--| ice-cream-making (e.g
--| lowering the temperature of the fluids by using eg. the pot-freezer meth=
od:
--| the temperature of the ingredients is reduced by placing them inside a tu=
b
--| filled with ice and salt) Cf. http://en.wikipedia.org/wiki/Ice_cream .
--|
--| At the end: just only pointing to the long MSA thread on --|N2 gas-LN2|--
--| starting May 2nd 2009
--| Conc: [Microscopy] Nitrogen leak (JEOL 6701F SEM)
--|
--|
--| Regards,
--| Wolfgang MUSS
--| Head EM-Lab, Pathology
--| SALK-LKH (Gen. Hosp.)
--| SALZBURG Austria
--|
--|
--|
--|
--| --| -----Urspr=C3=83=C6=92=C3=82=C2=BCngliche Nachricht-----
--| --| Von: richard.ross-at-allisontransmission.com
--| --| [mailto:richard.ross-at-allisontransmission.com]
--| --| Gesendet: Dienstag, 26. J=C3=83=C6=92=C3=82=C2=A4nner 2010 14:57
--| --| An: Mu=C3=83=C6=92=C3=85=C2=B8 Wolfgang
--| --| Betreff: [Microscopy] Re: Liquid Nitrogen Safety [Ice-Cream made with
--| --| LN2, concerns about]
--| --|
--| --|
--| -------------------------------------------------------------------------=
-
--| --| The Microscopy ListServer -- CoSponsor: The Microscopy Society of
--| America
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--| --|
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---
--| --| Way back when, Chuck Fiori used to put on a demo with LN2 at the LeHigh
--| SEM school.
--| Chuck wanted to show that sometimes its more hazardous to trap LN2 agains=
t
--| the body with protective gear than to give the liedenfrost effect room to
--| create its protective blanket of gas. If the LN2 was provided, he would
--| eventually take some into his mouth and blow 'smoke' rings. He told of a
--| time when he accidentally swallowed some of the LN2 and the resulting
--| stomach gas pressure caused him to black out, falling to the stage
--| unconscious.
--| Ultimately the excess pressure relieved itself as a belch and Chuck came
--| to.
--| Chuck certainly taught me things in ways that I remember! |--
--| --|
--| --| =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
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--| 27, 37 -- Subject: [Microscopy] Re: Liquid Nitrogen Safety
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--=20
"America believes in education; the average professor earns more money in a
year than a professional athlete earns in a whole week." Evan Esar

--0016e6d78426304381047e19d3a8
Content-Type: text/html; charset=UTF-8
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I prefer the hand-cranked version, myself. =C2=A0Not only does it give more=
direct tactile feedback on the overall thickness of the ice cream, but it =
gives the baby (To me) cousins something to do that is out of the way of my=
kitchen chaos on Thanksgiving!|--div--|
|--br--||--/div--||--div--|As for the health aspect, there was a study
that I read with=
in the last three or four years (I can't remember anything specific) th=
at said that the largest contributor to weight gain when eating is the ment=
al state of the eater. =C2=A0I was discussing this with a colleague of mine=
and we came to the conclusion that it must be true, based on the French. =
=C2=A0(My apologies at this generalization, but I am basing my experience o=
n the four incredible weeks I spent wandering around the French countryside=
...) =C2=A0|--/div--|
|--div--||--br--||--/div--||--div--|The French eat wonderfully
decadent, flavorful, and ric=
h foods, and drink gallons of truly elegant wine. =C2=A0Neither me nor my c=
olleague remembered seeing any obese French people (We're sure they exi=
st, but they don't seem to be the norm as it seems in the US). =C2=A0We=
also experienced the wonderful French culture of taking your time while ea=
ting, enjoying both the meal and the company, not rushing through it.|--/div--|
|--div--||--br--||--/div--||--div--|We could also come up with more
French nationals who li=
ved to respectable old age than we could of any other
nationality.|--/div--||--di=
v--||--br--||--/div--||--div--|I know that this is in no way a
scientific observation, bu=
t it's still enough to make me want to wander around rural France for t=
he rest of my life eating hand-churned ice cream...|--/div--|
|--div--||--br--||--/div--||--div--|Great. =C2=A0Now I'm hungry.
=C2=A0First time this =
list has done that to
me...|--/div--||--div--||--br--||--/div--||--div--|--Justin A.
Kraft|--/div=
--||--div--|Professional Lab Rat/High Energy Physics Server
Monkey|--/div--||--div--|Flor=
ida International University|--/div--|
|--div--|Miami, FL|--br--||--br--||--div class=3D"gmail_quote"--|On
Tue, Jan 26, 2010 at 6:=
19 PM, |--span dir=3D"ltr"--|<|--a
href=3D"mailto:kenconverse-at-qualityimages.b=
iz"--|kenconverse-at-qualityimages.biz|--/a--|>|--/span--|
wrote:|--br--||--blockquote clas=
s=3D"gmail_quote" style=3D"margin:0 0 0 .8ex;border-left:1px #ccc solid;pad=
ding-left:1ex;"--|
|--div class=3D"im"--||--br--|
|--br--|
|--br--|
---------------------------------------------------------------------------=
-|--br--|
The Microscopy ListServer -- CoSponsor: =C2=A0The Microscopy Society of Ame=
rica|--br--|
To =C2=A0Subscribe/Unsubscribe -- |--a href=3D"http://www.microscopy.com/Micr=
oscopyListserver
On-Line" target=3D"_blank"--|http://www.microscopy.com/MicroscopyListserver|--b=
r--|
On-Line|--/a--| Help |--a
href=3D"http://www.microscopy.com/MicroscopyListserver/=
FAQ.html" target=3D"_blank"--|http://www.microscopy.com/MicroscopyListserver/=
FAQ.html|--/a--||--br--|
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-|--br--|
|--br--|
|--/div--|Don,|--br--|
You're right! =C2=A0It's a dirty job, but somebody's got to do =
it. =C2=A0My wife and I got an electric ice cream maker as a wedding gift t=
hat we haven't used nearly enough. =C2=A0My favorite recipe not only in=
cludes the heavy cream, but a half dozen eggs. =C2=A0To die for...|--br--|

|--br--|
Of course the kill-joys will tell us that the heavy cream, the cholesterol =
in the eggs and all the sugar will kill us, not to mention the environmenta=
l effects of all the cattle and chickens, plus the inhumane conditions they=
are kept in.|--br--|

|--br--|
I'll have another serving of that custard ice cream recipe, thank you.|--=
br--|
|--div class=3D"im"--||--br--|
Ken Converse|--br--|
owner|--br--|
|--br--|
QUALITY IMAGES|--br--|
Servicing Scanning Electron Microscopes|--br--|
Since 1981|--br--|
474 So. Bridgton Rd.|--br--|
Bridgton, ME =C2=A004009|--br--|
207-647-4348|--br--|
Fax 207-647-2688|--br--|
|--a href=3D"mailto:kenconverse-at-qualityimages.biz"--|kenconverse-at-qualityimages.=
biz|--/a--||--br--|
|--a href=3D"http://qualityimages.biz"
target=3D"_blank"--|qualityimages.biz|--/a=
--||--br--|
|--br--|
|--br--|
-----Original Message-----|--br--|
|--/div--||--div class=3D"im"--|X-from: |--a
href=3D"mailto:donald.gibbon-at-matcoinc.co=
m"--|donald.gibbon-at-matcoinc.com|--/a--| [mailto:|--a
href=3D"mailto:donald.gibbon-at-m=
atcoinc.com"--|donald.gibbon-at-matcoinc.com|--/a--|]|--br--|
Sent: Tuesday, January 26, 2010 5:34 PM|--br--|
To: |--a href=3D"mailto:kenconverse-at-qualityimages.biz"--|kenconverse-at-qualityima=
ges.biz|--/a--||--br--|
|--/div--||--div class=3D"im"--|Subject: [Microscopy] FW: Re: Liquid
Nitrogen Safet=
y|--br--|
|--br--|
|--br--|
|--br--|
|--br--|
---------------------------------------------------------------------------=
-|--br--|
The Microscopy ListServer -- CoSponsor: =C2=A0The Microscopy Society of Ame=
rica|--br--|
To =C2=A0Subscribe/Unsubscribe -- |--a href=3D"http://www.microscopy.com/Micr=
oscopyListserver
On-Line" target=3D"_blank"--|http://www.microscopy.com/MicroscopyListserver|--b=
r--|
On-Line|--/a--| Help |--a
href=3D"http://www.microscopy.com/MicroscopyListserver/=
FAQ.html" target=3D"_blank"--|http://www.microscopy.com/MicroscopyListserver/=
FAQ.html|--/a--||--br--|
---------------------------------------------------------------------------=
-|--br--|
|--br--|
|--br--|
|--br--|
|--br--|
|--/div--|As a semi-professional ice cream maker, I'd like to second Wolfga=
ng Muss's reference to the old-fashioned but utterly reliable and safe =
method of using a hand-cranked ice cream freezer rather than any other opti=
on. There are reasons why this works well, which, if ignored, explain why i=
t doesn't work sometimes! Making ice cream is an exercise in heat trans=
fer and crystal growth. The heat transfer surface is the metal wall of the =
tub which is rotated by the crank. Counter-rotating inside the tube is a pa=
ir of wooden paddles, keeping that surface clean and free of ice, constantl=
y removing the freezing cream. The idea is to get a eutectic mixture of sal=
t and ice on the outside, i.e., minimum possible temperature in this system=
, dump in the cream and start cranking. Don't ever stop for even one se=
cond. i.e., keep that surface clean, and in six-eight minutes your job will=
be done. The idea is to maximize the number of crystals and minimize their=
size. It is NOT a tedious process, !|--br--|

=C2=A0hardly enough to get winded.|--br--|
|--br--|
My most sought-after publication in my entire life was my paper on The Ther=
modynamics of Homemade Ice Cream, published in the Journal Of Chemical Educ=
ation. I still have the thermocouple-fitted ice cream freezer in which we m=
ade many gallons of ice cream, doing the experimental work for this paper. =
Somebody had to do it! And sticking your finger in the ice cream will not h=
arm you at all!|--br--|

|--br--|
Donald L. Gibbon|--br--|
|--div class=3D"im"--||--br--|
-----Original Message-----|--br--|
X-from: |--a href=3D"mailto:W.Muss-at-salk.at"--|W.Muss-at-salk.at|--/a--|
[mailto:|--a hre=
f=3D"mailto:W.Muss-at-salk.at"--|W.Muss-at-salk.at|--/a--|]|--br--|
Sent: Tuesday, January 26, 2010 12:23 PM|--br--|
To: Gibbon, Donald L.|--br--|

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: Robert.Brandom-at-Bruker-AXS.com
Name: Robert Brandom

Organization: New England Society for Microscopy

Title-Subject: [Filtered] NESM -- February 9 Dinner Meeting

Message: Greetings to all:

The NESM February Meeting is set for Tuesday evening, February 9, 2010.

This first of 4 NESM events for 2010 is being
hosted by Bruker Corporation at their facility in
Billerica, MA.

The preliminary agenda is as follows:

5:45-6:15 PM Registration & Bruker Open House

6:15-7:25 PM Buffet Supper (Open House continues)

7:25 PM Introductory Remarks - Warren MoberlyChan, 2010 NESM President

7:30-9:00 PM Technical Presentations

ìSpectroscopy in the Microscopy Worldî
Thomas J Tague Jr., Ph.D.
Bruker Optics, Inc.

ìDirect Correlation Between Optical and
Structural Properties on the Nanoscale:
Cathodoluminescence in Scanning Transmission Electron Microscopy ì
Silvija GradeËak, Ph.D.
Department of Materials Science and Engineering
Massachusetts Institute of Technology

9:00 PM Adjourn


The charge for this meeting will be collected at the event as follows:

} } NESM Members - $20
} } Students and retirees and members between
} } jobs - $10 } } Non-Members - $40 (this will
} } include a discounted membership for 2010)
} } The above rates will apply to all those who RSVP in Advance
} } All walk-ins who do not RSVP will be charged an additional $5


Please RSVP via email to nesm67-at-charter.net by Friday, Feb 5, 2010.

Full details including speaker abstract and bios,
map and directions to Bruker can be found at the
NESM website.

http://nesm.cims.harvard.edu/NESM_04.htm

We look forward to seeing you in Billerica !!


Bruker Corporation (meeting at Bruker Daltonics
building) 40 Manning Road Billerica, MA 01821

Need help en-route ñ call Robert Brandom (978) 257-3122

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From: apeer-at-halifax.k12.va.us
Date: Tue, 26 Jan 2010 19:41:53 -0600
Subject: [Microscopy] viaWWW: QX3 and QX5 Computer Microscope

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Email: apeer-at-halifax.k12.va.us
Name: Aleacia Peer

Organization: CSES

Title-Subject: [Filtered] QX3 and QX5 Computer Microscope

Message: I teach 5th grade and am incorporating the use of USB
computer microscopes into my classes.

I need to know where to buy the replacement bulbs for the Intel Play
QX3 and the Digital Blue QX5 computer microscopes.

I removed a bulb from one of the microscopes and the only marking on
it was 1.5W---no other information to help me find a replacement.

The Digital Blue website sells them, but they have been "Sold Out"
for quite some time.

Any help would be greatly appreciated!

Thanks!

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From: kraftpiano-at-gmail.com
Date: Wed, 27 Jan 2010 08:34:55 -0600
Subject: [Microscopy] Liquid Nitrogen Safety

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

Dear Justin,

I am happy that you enjoyed the french way-of-life.
I think that, rather the mental state, the amount eaten probably plays a critical role in weight gain.
Actually french enjoy tasteful meals because they eat for the pleasure, so they prefer to eat less but better.
The same comes with the beer between Belgium and Germany f.e.: the belgian beer comes in 25cl while the german in 0,5L flasks.
Eat slowly also helps the digestion. It is both a question of psychical and physical pleasure.
This makes me almost forget that I am not french. Well all the same...

Stephane

PS: why do we start a discussion about how to make ice-cream when there is -15°C outside!!


----- Original Message ----
X-from: "kraftpiano-at-gmail.com" {kraftpiano-at-gmail.com}
To: nizets2-at-yahoo.com
Sent: Wed, January 27, 2010 12:51:07 AM

I prefer the hand-cranked version, myself.  Not only does it give more
direct tactile feedback on the overall thickness of the ice cream, but it
gives the baby (To me) cousins something to do that is out of the way of my
kitchen chaos on Thanksgiving!

As for the health aspect, there was a study that I read within the last
three or four years (I can't remember anything specific) that said that the
largest contributor to weight gain when eating is the mental state of the
eater.  I was discussing this with a colleague of mine and we came to the
conclusion that it must be true, based on the French.  (My apologies at this
generalization, but I am basing my experience on the four incredible weeks I
spent wandering around the French countryside...)

The French eat wonderfully decadent, flavorful, and rich foods, and drink
gallons of truly elegant wine.  Neither me nor my colleague remembered
seeing any obese French people (We're sure they exist, but they don't seem
to be the norm as it seems in the US).  We also experienced the wonderful
French culture of taking your time while eating, enjoying both the meal and
the company, not rushing through it.

We could also come up with more French nationals who lived to respectable
old age than we could of any other nationality.

I know that this is in no way a scientific observation, but it's still
enough to make me want to wander around rural France for the rest of my life
eating hand-churned ice cream...

Great.  Now I'm hungry.  First time this list has done that to me...

--Justin A. Kraft
Professional Lab Rat/High Energy Physics Server Monkey
Florida International University
Miami, FL


--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar

n Tue, Jan 26, 2010 at 6:19 PM, |--kenconverse-at-qualityimages.biz--| wrote:

--|
--|
--|
--|
--| -------------------------------------------------------------------------=
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--|
--| Don,
--| You're right!  It's a dirty job, but somebody's got to do it.  My wife an=
d
--| I got an electric ice cream maker as a wedding gift that we haven't used
--| nearly enough.  My favorite recipe not only includes the heavy cream, but=
a
--| half dozen eggs.  To die for...
--|
--| Of course the kill-joys will tell us that the heavy cream, the cholestero=
l
--| in the eggs and all the sugar will kill us, not to mention the environmen=
tal
--| effects of all the cattle and chickens, plus the inhumane conditions they
--| are kept in.
--|
--| I'll have another serving of that custard ice cream recipe, thank you.
--|
--| Ken Converse
--| owner
--|
--| QUALITY IMAGES
--| Servicing Scanning Electron Microscopes
--| Since 1981
--| 474 So. Bridgton Rd.
--| Bridgton, ME  04009
--| 207-647-4348
--| Fax 207-647-2688
--| kenconverse-at-qualityimages.biz
--| qualityimages.biz
--|
--|
--| -----Original Message-----
--| X-from: donald.gibbon-at-matcoinc.com [mailto:donald.gibbon-at-matcoinc.com]
--| Sent: Tuesday, January 26, 2010 5:34 PM
--| To: kenconverse-at-qualityimages.biz
--| Subject: [Microscopy] FW: Re: Liquid Nitrogen Safety
--|
--|
--|
--|
--|
--| -------------------------------------------------------------------------=
---
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a
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---
--|
--|
--|
--|
--| As a semi-professional ice cream maker, I'd like to second Wolfgang Muss'=
s
--| reference to the old-fashioned but utterly reliable and safe method of us=
ing
--| a hand-cranked ice cream freezer rather than any other option. There are
--| reasons why this works well, which, if ignored, explain why it doesn't wo=
rk
--| sometimes! Making ice cream is an exercise in heat transfer and crystal
--| growth. The heat transfer surface is the metal wall of the tub which is
--| rotated by the crank. Counter-rotating inside the tube is a pair of woode=
n
--| paddles, keeping that surface clean and free of ice, constantly removing =
the
--| freezing cream. The idea is to get a eutectic mixture of salt and ice on =
the
--| outside, i.e., minimum possible temperature in this system, dump in the
--| cream and start cranking. Don't ever stop for even one second. i.e., keep
--| that surface clean, and in six-eight minutes your job will be done. The i=
dea
--| is to maximize the number of crystals and minimize their size. It is NOT =
a
--| tedious process, !
--|  hardly enough to get winded.
--|
--| My most sought-after publication in my entire life was my paper on The
--| Thermodynamics of Homemade Ice Cream, published in the Journal Of Chemica=
l
--| Education. I still have the thermocouple-fitted ice cream freezer in whic=
h
--| we made many gallons of ice cream, doing the experimental work for this
--| paper. Somebody had to do it! And sticking your finger in the ice cream w=
ill
--| not harm you at all!
--|
--| Donald L. Gibbon
--|
--| -----Original Message-----
--| X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at]
--| Sent: Tuesday, January 26, 2010 12:23 PM
--| To: Gibbon, Donald L.
--| Subject: [Microscopy] Re: Liquid Nitrogen Safety
--|
--|
--|
--|
--|
--| -------------------------------------------------------------------------=
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--| http://www.microscopy.com/MicroscopyListserver
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--|
--| Good morning, good afternoon, good evening - hopefully any somewhere will
--| apply,
--| hello, dear colleagues,
--|
--| Just to add my 2 =C3=83=C2=A2=C3=A2=E2=82=AC=C5=A1=C3=82=C2=AC-cents (apo=
logize if the message became too long):
--|
--| Yes, we can discuss about children STICK THEIR FINGERS IN THE LIQUID
--| NITROGEN and other stuff....  in general,
--| since in EM we sometimes have to work with 'cruel' things and substances
--| (and others a little bit more often).
--| Cryogenics also CAN be hazardous (as --|hot|-- water is/can be),
depending on
--| how we are --|working with|-- it.
--| If we couldn't do that safe(ly) it necessarily / perhaps would have been
--| forbidden for a long time.
--|
--| There are a lot of sources on the web one can find concerning use and
--| misuse of e. g. liquid nitrogen:
--| for example:
--|
--| http://www.altair.org/hazard.html  (DONT DIE ! Laboratory Hazards: Safety=
,
--| Prevention, First Aid, C 2001-2010)
--|
--|
--| http://www.reachoutmichigan.org/funexperiments/agesubject/lessons/nitroge=
n.html
--| ( "Fun experiments" website last updated 2003)
--|
--|
--| http://smb.slac.stanford.edu/users_guide/manual/Experiment_policies.html#=
SECTION00027000000000000000
--| (describes hazards and proper handling procedures for work with liquid
--| nitrogen, 2010)
--|
--| http://stores.biochem.uiowa.edu/Pages/ln2msds.htm
--| ("old" but IMO "good" example for a US-Univ's Safety Data Sheet)
--|
--|
--| http://www.chaosscience.org.uk/dem/public_html//article.php?story=3D20031=
216175107931
--| (Liquid Nitrogen Risk Assessment, 16/12/03 )
--|
--| Also, some Labs are questioning in advance for safety issues on the thing=
s
--| someone will bring into the Lab:
--| e.g. http://www.sbc.anl.gov/pdfs/hazard_assessment_form.pdf  (take a look
--| for 'cryogens', Argonne Natl. Lab, SBC Hazard Assessment Form), perhaps j=
ust
--| to be on the safe side for their own staff people and Lab's interior.
--|
--| AND finally, YES, LN2-ingestion - to take a mouthfull of LN2 and blast it
--| off - only for the EXPERIENCED CAN BE a junky-funky trick, BUT -
--| on the other side - for the unexperienced (unfortunately also for the
--| 'careless' experienced I have to admit!) - it could be not only dangerous
--| but LIFE threatening:  cf.
--| http://pediatrics.aappublications.org/cgi/reprint/105/1/121
--| ( Benjamin Z. Koplewitz, et al....  Gastric Perforation Attributable to
--| Liquid Nitrogen Ingestion,
--| in:  Pediatrics 2000;105;121-123, open Access)
--|
--| A similar situation has been reported on
--| http://www.darwinawards.com/personal/personal2000-25.html,
--| which eventually was --|awarded|-- with the DARWIN AWARD in 2000
(=3D=3D--|cit:=
"The
--| Darwin Awards salute the improvement of the human genome by honoring thos=
e
--| who accidentally remove themselves from it..." end of cit.)
--|
--| I personally have no doubt that it IS possible (and I have done that for
--| demonstration of "Leidenfrost phenomenon" a lot of times way back...) to
--| place one or two fingers, eventually(and forsure) the whole hand into liq=
uid
--| nitrogen for 1-2 seconds, PROVIDED some necessary PREREQUISITES which - I=
N
--| ADVANCE - have to be theoretically explained and are to be met to/for tho=
se
--| --|trying|-- such an "adventure" and IMO only should be done in a SUPERVISED
--| situation.
--|
--| Despite (or better: BECAUSE) respecting all those "little" |--problem-maker=
s--|
--| in our daily work
--| (if this is not done by other staff members, I have to to do such work
--| literally by myself) I am not in a blue funk of perhaps desastrous result=
s
--| of using/handling all those substances like
--| OsO4 (hazard, toxic), p-phenylenediamine (pesticide, toxic), uranyl-aceta=
te
--| from stock powder (radioactive), cryogens (like LN2, precooled isobutane,
--| also Freon gas [some residual inventory] if the pressurized can will be u=
sed
--| inverted) and other substances/chemicals (mostly used in very small
--| quantities), not to forget all those "may be hazardous resins" out in the
--| dark...
--|
--| WHY I am NOT AFRAID/do not fear those:  since I have learned - before
--| handling and using those -
--| WHAT the respective properties of materials/substances/fluids are,
--| HOW they have ( practically ) to be used safely,
--| WHAT the consequences eventually will be (personal and for others) if
--| handled unsafe, and
--| HOW to dispose of (remnants, by-products of reactions, etc.) properly.
--|
--| (as an example for awareness: read:
--| http://www.hull.ac.uk/chemstores/coshhadv.html (COSHH- Control of
--| Substances Hazardous to Health, Univ.Hull, UK:  'part of the U.K.'s CRIMI=
NAL
--| LAW' !)
--|
--| Anybody who has |--studied--| properties of chemicals etc., their
handling an=
d
--| use properly and correctly will know how to proceed with the use of mater=
ial
--| and substances we use in an EM-lab. If this has not be done, no one shoul=
d
--| do --|things|-- he never got explained and does not know about
consequences i=
n
--| case of misuse.
--|
--| There IMO is no need to disallow or eventually prohibit (necessary) actio=
ns
--| which to the unexperienced (perhaps) will be harmful, but to the experien=
ced
--| does not apply seriously.
--| So, last but not least: it is the responsibilty of the "experienced" to
--| teach the unexperienced... so you as the |--experienced--| decide
what can be
--| done/shown and what CANNOT be done /shown.
--|
--| Concerning: Ice-Cream-experiments with LN2, those perhaps are an
--| "interesting" demonstration for a new practical application but easily
--| can/could be safely substituted by |--old fashioned--| ice-cream-making (e.g
--| lowering the temperature of the fluids by using eg. the  pot-freezer meth=
od:
--| the temperature of the ingredients is reduced by placing them inside a tu=
b
--| filled with ice and salt) Cf. http://en.wikipedia.org/wiki/Ice_cream .
--|
--| At the end: just only pointing to the long MSA thread on --|N2 gas-LN2|--
--| starting May 2nd 2009
--| Conc: [Microscopy] Nitrogen leak (JEOL 6701F SEM)
--|
--|
--| Regards,
--| Wolfgang MUSS
--| Head EM-Lab, Pathology
--| SALK-LKH (Gen. Hosp.)
--| SALZBURG Austria
--|
--|
--|
--|
--| --| -----Urspr=C3=83=C6=92=C3=82=C2=BCngliche Nachricht-----
--| --| Von: richard.ross-at-allisontransmission.com
--| --| [mailto:richard.ross-at-allisontransmission.com]
--| --| Gesendet: Dienstag, 26. J=C3=83=C6=92=C3=82=C2=A4nner 2010 14:57
--| --| An: Mu=C3=83=C6=92=C3=85=C2=B8 Wolfgang
--| --| Betreff: [Microscopy] Re: Liquid Nitrogen Safety [Ice-Cream made with
--| --| LN2, concerns about]
--| --|
--| --|
--| -------------------------------------------------------------------------=
-
--| --| The Microscopy ListServer -- CoSponsor:  The Microscopy Society of
--| America
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--| --|
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---
--| --| Way back when, Chuck Fiori used to put on a demo with LN2 at the LeHigh
--| SEM school.
--| Chuck wanted to show that sometimes its more hazardous to trap LN2 agains=
t
--| the body with protective gear than to give the liedenfrost effect room to
--| create its protective blanket of gas. If the LN2 was provided, he would
--| eventually take some into his mouth and blow 'smoke' rings. He told of a
--| time when he accidentally swallowed some of the LN2 and the resulting
--| stomach gas pressure caused him to black out, falling to the stage
--| unconscious.
--| Ultimately the excess pressure relieved itself as a belch and Chuck came
--| to.
--| Chuck certainly taught me things in ways that I remember! |--
--| --|
--| --| =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
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=3D=3D=3D=3D=3D=3D=3D=3D
--| --| 1, 25 -- From richard.ross-at-allisontransmission.com Tue Jan 26 07:53:09
--| 2010
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--| --| 1, 25 -- X-MIMETrack: Serialize by Router on
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--=20
"America believes in education; the average professor earns more money in a
year than a professional athlete earns in a whole week." Evan Esar

--0016e6d78426304381047e19d3a8
Content-Type: text/html; charset=UTF-8
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I prefer the hand-cranked version, myself. =C2=A0Not only does it give more=
direct tactile feedback on the overall thickness of the ice cream, but it =
gives the baby (To me) cousins something to do that is out of the way of my=
kitchen chaos on Thanksgiving!|--div--|
|--br--||--/div--||--div--|As for the health aspect, there was a study
that I read with=
in the last three or four years (I can't remember anything specific) th=
at said that the largest contributor to weight gain when eating is the ment=
al state of the eater. =C2=A0I was discussing this with a colleague of mine=
and we came to the conclusion that it must be true, based on the French. =
=C2=A0(My apologies at this generalization, but I am basing my experience o=
n the four incredible weeks I spent wandering around the French countryside=
...) =C2=A0|--/div--|
|--div--||--br--||--/div--||--div--|The French eat wonderfully
decadent, flavorful, and ric=
h foods, and drink gallons of truly elegant wine. =C2=A0Neither me nor my c=
olleague remembered seeing any obese French people (We're sure they exi=
st, but they don't seem to be the norm as it seems in the US). =C2=A0We=
also experienced the wonderful French culture of taking your time while ea=
ting, enjoying both the meal and the company, not rushing through it.|--/div--|
|--div--||--br--||--/div--||--div--|We could also come up with more
French nationals who li=
ved to respectable old age than we could of any other
nationality.|--/div--||--di=
v--||--br--||--/div--||--div--|I know that this is in no way a
scientific observation, bu=
t it's still enough to make me want to wander around rural France for t=
he rest of my life eating hand-churned ice cream...|--/div--|
|--div--||--br--||--/div--||--div--|Great. =C2=A0Now I'm hungry.
=C2=A0First time this =
list has done that to
me...|--/div--||--div--||--br--||--/div--||--div--|--Justin A.
Kraft|--/div=
--||--div--|Professional Lab Rat/High Energy Physics Server
Monkey|--/div--||--div--|Flor=
ida International University|--/div--|
|--div--|Miami, FL|--br--||--br--||--div class=3D"gmail_quote"--|On
Tue, Jan 26, 2010 at 6:=
19 PM,  |--span dir=3D"ltr"--|<|--a
href=3D"mailto:kenconverse-at-qualityimages.b=
iz"--|kenconverse-at-qualityimages.biz|--/a--|>|--/span--|
wrote:|--br--||--blockquote clas=
s=3D"gmail_quote" style=3D"margin:0 0 0 .8ex;border-left:1px #ccc solid;pad=
ding-left:1ex;"--|
|--div class=3D"im"--||--br--|
|--br--|
|--br--|
---------------------------------------------------------------------------=
-|--br--|
The Microscopy ListServer -- CoSponsor: =C2=A0The Microscopy Society of Ame=
rica|--br--|
To =C2=A0Subscribe/Unsubscribe -- |--a href=3D"http://www.microscopy.com/Micr=
oscopyListserver
On-Line" target=3D"_blank"--|http://www.microscopy.com/MicroscopyListserver|--b=
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On-Line|--/a--| Help |--a
href=3D"http://www.microscopy.com/MicroscopyListserver/=
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-|--br--|
|--br--|
|--/div--|Don,|--br--|
You're right! =C2=A0It's a dirty job, but somebody's got to do =
it. =C2=A0My wife and I got an electric ice cream maker as a wedding gift t=
hat we haven't used nearly enough. =C2=A0My favorite recipe not only in=
cludes the heavy cream, but a half dozen eggs. =C2=A0To die for...|--br--|

|--br--|
Of course the kill-joys will tell us that the heavy cream, the cholesterol =
in the eggs and all the sugar will kill us, not to mention the environmenta=
l effects of all the cattle and chickens, plus the inhumane conditions they=
are kept in.|--br--|

|--br--|
I'll have another serving of that custard ice cream recipe, thank you.|--=
br--|
|--div class=3D"im"--||--br--|
Ken Converse|--br--|
owner|--br--|
|--br--|
QUALITY IMAGES|--br--|
Servicing Scanning Electron Microscopes|--br--|
Since 1981|--br--|
474 So. Bridgton Rd.|--br--|
Bridgton, ME =C2=A004009|--br--|
207-647-4348|--br--|
Fax 207-647-2688|--br--|
|--a href=3D"mailto:kenconverse-at-qualityimages.biz"--|kenconverse-at-qualityimages.=
biz|--/a--||--br--|
|--a href=3D"http://qualityimages.biz"
target=3D"_blank"--|qualityimages.biz|--/a=
--||--br--|
|--br--|
|--br--|
-----Original Message-----|--br--|
|--/div--||--div class=3D"im"--|X-from: |--a
href=3D"mailto:donald.gibbon-at-matcoinc.co=
m"--|donald.gibbon-at-matcoinc.com|--/a--| [mailto:|--a
href=3D"mailto:donald.gibbon-at-m=
atcoinc.com"--|donald.gibbon-at-matcoinc.com|--/a--|]|--br--|
Sent: Tuesday, January 26, 2010 5:34 PM|--br--|
To: |--a href=3D"mailto:kenconverse-at-qualityimages.biz"--|kenconverse-at-qualityima=
ges.biz|--/a--||--br--|
|--/div--||--div class=3D"im"--|Subject: [Microscopy] FW: Re: Liquid
Nitrogen Safet=
y|--br--|
|--br--|
|--br--|
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To =C2=A0Subscribe/Unsubscribe -- |--a href=3D"http://www.microscopy.com/Micr=
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On-Line|--/a--| Help |--a
href=3D"http://www.microscopy.com/MicroscopyListserver/=
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-|--br--|
|--br--|
|--br--|
|--br--|
|--br--|
|--/div--|As a semi-professional ice cream maker, I'd like to second Wolfga=
ng Muss's reference to the old-fashioned but utterly reliable and safe =
method of using a hand-cranked ice cream freezer rather than any other opti=
on. There are reasons why this works well, which, if ignored, explain why i=
t doesn't work sometimes! Making ice cream is an exercise in heat trans=
fer and crystal growth. The heat transfer surface is the metal wall of the =
tub which is rotated by the crank. Counter-rotating inside the tube is a pa=
ir of wooden paddles, keeping that surface clean and free of ice, constantl=
y removing the freezing cream. The idea is to get a eutectic mixture of sal=
t and ice on the outside, i.e., minimum possible temperature in this system=
, dump in the cream and start cranking. Don't ever stop for even one se=
cond. i.e., keep that surface clean, and in six-eight minutes your job will=
be done. The idea is to maximize the number of crystals and minimize their=
size. It is NOT a tedious process, !|--br--|

=C2=A0hardly enough to get winded.|--br--|
|--br--|
My most sought-after publication in my entire life was my paper on The Ther=
modynamics of Homemade Ice Cream, published in the Journal Of Chemical Educ=
ation. I still have the thermocouple-fitted ice cream freezer in which we m=
ade many gallons of ice cream, doing the experimental work for this paper. =
Somebody had to do it! And sticking your finger in the ice cream will not h=
arm you at all!|--br--|

|--br--|
Donald L. Gibbon|--br--|
|--div class=3D"im"--||--br--|
-----Original Message-----|--br--|
X-from: |--a href=3D"mailto:W.Muss-at-salk.at"--|W.Muss-at-salk.at|--/a--|
[mailto:|--a hre=
f=3D"mailto:W.Muss-at-salk.at"--|W.Muss-at-salk.at|--/a--|]|--br--|
Sent: Tuesday, January 26, 2010 12:23 PM|--br--|
To: Gibbon, Donald L.|--br--|

I agree that enjoying the meal and smaller portions probably are the
key... As to the discussion on ice cream, well, I somehow (I keep
asking myself why) ended up in south Florida, where it is usually
never below about 60 degrees F...

While we are on the subject of food, though, since everyone on this
list is scientifically minded, has anyone else found that they treat
cooking like a science? My personal forte is baking, but the methods
I use in the kitchen are very similar to methods used in the lab...
After all, a recipe is merely a sustenance production protocol, right?

--Justin.

On Wed, Jan 27, 2010 at 7:25 AM, Stephane Nizet {nizets2-at-yahoo.com} wrote:
} Dear Justin,
}
} I am happy that you enjoyed the french way-of-life.
} I think that, rather the mental state, the amount eaten probably plays a critical role in weight gain.
} Actually french enjoy tasteful meals because they eat for the pleasure, so they prefer to eat less but better.
} The same comes with the beer between Belgium and Germany f.e.: the belgian beer comes in 25cl while the german in 0,5L flasks.
} Eat slowly also helps the digestion. It is both a question of psychical and physical pleasure.
} This makes me almost forget that I am not french. Well all the same...
}
} Stephane
}
} PS: why do we start a discussion about how to make ice-cream when there is -15°C outside!!
}
}
} ----- Original Message ----
} From: "kraftpiano-at-gmail.com" {kraftpiano-at-gmail.com}
} To: nizets2-at-yahoo.com
} Sent: Wed, January 27, 2010 12:51:07 AM
} Subject: [Microscopy] Re: Liquid Nitrogen Safety
}
}
}
}
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} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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}
} I prefer the hand-cranked version, myself.  Not only does it give more
} direct tactile feedback on the overall thickness of the ice cream, but it
} gives the baby (To me) cousins something to do that is out of the way of my
} kitchen chaos on Thanksgiving!
}
} As for the health aspect, there was a study that I read within the last
} three or four years (I can't remember anything specific) that said that the
} largest contributor to weight gain when eating is the mental state of the
} eater.  I was discussing this with a colleague of mine and we came to the
} conclusion that it must be true, based on the French.  (My apologies at this
} generalization, but I am basing my experience on the four incredible weeks I
} spent wandering around the French countryside...)
}
} The French eat wonderfully decadent, flavorful, and rich foods, and drink
} gallons of truly elegant wine.  Neither me nor my colleague remembered
} seeing any obese French people (We're sure they exist, but they don't seem
} to be the norm as it seems in the US).  We also experienced the wonderful
} French culture of taking your time while eating, enjoying both the meal and
} the company, not rushing through it.
}
} We could also come up with more French nationals who lived to respectable
} old age than we could of any other nationality.
}
} I know that this is in no way a scientific observation, but it's still
} enough to make me want to wander around rural France for the rest of my life
} eating hand-churned ice cream...
}
} Great.  Now I'm hungry.  First time this list has done that to me...
}
} --Justin A. Kraft
} Professional Lab Rat/High Energy Physics Server Monkey
} Florida International University
} Miami, FL
}
}
} --
} "America believes in education; the average professor earns more money
} in a year than a professional athlete earns in a whole week." Evan
} Esar
}
} n Tue, Jan 26, 2010 at 6:19 PM, |--kenconverse-at-qualityimages.biz--| wrote:
}
} --|
} --|
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} --|
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} ---
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} --|
} --| Don,
} --| You're right!  It's a dirty job, but somebody's got to do it.  My wife an=
} d
} --| I got an electric ice cream maker as a wedding gift that we haven't used
} --| nearly enough.  My favorite recipe not only includes the heavy cream, but=
} a
} --| half dozen eggs.  To die for...
} --|
} --| Of course the kill-joys will tell us that the heavy cream, the cholestero=
} l
} --| in the eggs and all the sugar will kill us, not to mention the environmen=
} tal
} --| effects of all the cattle and chickens, plus the inhumane conditions they
} --| are kept in.
} --|
} --| I'll have another serving of that custard ice cream recipe, thank you.
} --|
} --| Ken Converse
} --| owner
} --|
} --| QUALITY IMAGES
} --| Servicing Scanning Electron Microscopes
} --| Since 1981
} --| 474 So. Bridgton Rd.
} --| Bridgton, ME  04009
} --| 207-647-4348
} --| Fax 207-647-2688
} --| kenconverse-at-qualityimages.biz
} --| qualityimages.biz
} --|
} --|
} --| -----Original Message-----
} --| X-from: donald.gibbon-at-matcoinc.com [mailto:donald.gibbon-at-matcoinc.com]
} --| Sent: Tuesday, January 26, 2010 5:34 PM
} --| To: kenconverse-at-qualityimages.biz
} --| Subject: [Microscopy] FW: Re: Liquid Nitrogen Safety
} --|
} --|
} --|
} --|
} --|
} --| -------------------------------------------------------------------------=
} ---
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} a
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} ---
} --|
} --|
} --|
} --|
} --| As a semi-professional ice cream maker, I'd like to second Wolfgang Muss'=
} s
} --| reference to the old-fashioned but utterly reliable and safe method of us=
} ing
} --| a hand-cranked ice cream freezer rather than any other option. There are
} --| reasons why this works well, which, if ignored, explain why it doesn't wo=
} rk
} --| sometimes! Making ice cream is an exercise in heat transfer and crystal
} --| growth. The heat transfer surface is the metal wall of the tub which is
} --| rotated by the crank. Counter-rotating inside the tube is a pair of woode=
} n
} --| paddles, keeping that surface clean and free of ice, constantly removing =
} the
} --| freezing cream. The idea is to get a eutectic mixture of salt and ice on =
} the
} --| outside, i.e., minimum possible temperature in this system, dump in the
} --| cream and start cranking. Don't ever stop for even one second. i.e., keep
} --| that surface clean, and in six-eight minutes your job will be done. The i=
} dea
} --| is to maximize the number of crystals and minimize their size. It is NOT =
} a
} --| tedious process, !
} --|  hardly enough to get winded.
} --|
} --| My most sought-after publication in my entire life was my paper on The
} --| Thermodynamics of Homemade Ice Cream, published in the Journal Of Chemica=
} l
} --| Education. I still have the thermocouple-fitted ice cream freezer in whic=
} h
} --| we made many gallons of ice cream, doing the experimental work for this
} --| paper. Somebody had to do it! And sticking your finger in the ice cream w=
} ill
} --| not harm you at all!
} --|
} --| Donald L. Gibbon
} --|
} --| -----Original Message-----
} --| X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at]
} --| Sent: Tuesday, January 26, 2010 12:23 PM
} --| To: Gibbon, Donald L.
} --| Subject: [Microscopy] Re: Liquid Nitrogen Safety
} --|
} --|
} --|
} --|
} --|
} --| -------------------------------------------------------------------------=
} ---
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} a
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} ---
} --|
} --| Good morning, good afternoon, good evening - hopefully any somewhere will
} --| apply,
} --| hello, dear colleagues,
} --|
} --| Just to add my 2 =C3=83=C2=A2=C3=A2=E2=82=AC=C5=A1=C3=82=C2=AC-cents (apo=
} logize if the message became too long):
} --|
} --| Yes, we can discuss about children STICK THEIR FINGERS IN THE LIQUID
} --| NITROGEN and other stuff....  in general,
} --| since in EM we sometimes have to work with 'cruel' things and substances
} --| (and others a little bit more often).
} --| Cryogenics also CAN be hazardous (as --|hot|-- water is/can be),
} depending on
} --| how we are --|working with|-- it.
} --| If we couldn't do that safe(ly) it necessarily / perhaps would have been
} --| forbidden for a long time.
} --|
} --| There are a lot of sources on the web one can find concerning use and
} --| misuse of e. g. liquid nitrogen:
} --| for example:
} --|
} --| http://www.altair.org/hazard.html  (DONT DIE ! Laboratory Hazards: Safety=
} ,
} --| Prevention, First Aid, C 2001-2010)
} --|
} --|
} --| http://www.reachoutmichigan.org/funexperiments/agesubject/lessons/nitroge=
} n.html
} --| ( "Fun experiments" website last updated 2003)
} --|
} --|
} --| http://smb.slac.stanford.edu/users_guide/manual/Experiment_policies.html#=
} SECTION00027000000000000000
} --| (describes hazards and proper handling procedures for work with liquid
} --| nitrogen, 2010)
} --|
} --| http://stores.biochem.uiowa.edu/Pages/ln2msds.htm
} --| ("old" but IMO "good" example for a US-Univ's Safety Data Sheet)
} --|
} --|
} --| http://www.chaosscience.org.uk/dem/public_html//article.php?story=3D20031=
} 216175107931
} --| (Liquid Nitrogen Risk Assessment, 16/12/03 )
} --|
} --| Also, some Labs are questioning in advance for safety issues on the thing=
} s
} --| someone will bring into the Lab:
} --| e.g. http://www.sbc.anl.gov/pdfs/hazard_assessment_form.pdf  (take a look
} --| for 'cryogens', Argonne Natl. Lab, SBC Hazard Assessment Form), perhaps j=
} ust
} --| to be on the safe side for their own staff people and Lab's interior.
} --|
} --| AND finally, YES, LN2-ingestion - to take a mouthfull of LN2 and blast it
} --| off - only for the EXPERIENCED CAN BE a junky-funky trick, BUT -
} --| on the other side - for the unexperienced (unfortunately also for the
} --| 'careless' experienced I have to admit!) - it could be not only dangerous
} --| but LIFE threatening:  cf.
} --| http://pediatrics.aappublications.org/cgi/reprint/105/1/121
} --| ( Benjamin Z. Koplewitz, et al....  Gastric Perforation Attributable to
} --| Liquid Nitrogen Ingestion,
} --| in:  Pediatrics 2000;105;121-123, open Access)
} --|
} --| A similar situation has been reported on
} --| http://www.darwinawards.com/personal/personal2000-25.html,
} --| which eventually was --|awarded|-- with the DARWIN AWARD in 2000
} (=3D=3D--|cit:=
} "The
} --| Darwin Awards salute the improvement of the human genome by honoring thos=
} e
} --| who accidentally remove themselves from it..." end of cit.)
} --|
} --| I personally have no doubt that it IS possible (and I have done that for
} --| demonstration of "Leidenfrost phenomenon" a lot of times way back...) to
} --| place one or two fingers, eventually(and forsure) the whole hand into liq=
} uid
} --| nitrogen for 1-2 seconds, PROVIDED some necessary PREREQUISITES which - I=
} N
} --| ADVANCE - have to be theoretically explained and are to be met to/for tho=
} se
} --| --|trying|-- such an "adventure" and IMO only should be done in a SUPERVISED
} --| situation.
} --|
} --| Despite (or better: BECAUSE) respecting all those "little" |--problem-maker=
} s--|
} --| in our daily work
} --| (if this is not done by other staff members, I have to to do such work
} --| literally by myself) I am not in a blue funk of perhaps desastrous result=
} s
} --| of using/handling all those substances like
} --| OsO4 (hazard, toxic), p-phenylenediamine (pesticide, toxic), uranyl-aceta=
} te
} --| from stock powder (radioactive), cryogens (like LN2, precooled isobutane,
} --| also Freon gas [some residual inventory] if the pressurized can will be u=
} sed
} --| inverted) and other substances/chemicals (mostly used in very small
} --| quantities), not to forget all those "may be hazardous resins" out in the
} --| dark...
} --|
} --| WHY I am NOT AFRAID/do not fear those:  since I have learned - before
} --| handling and using those -
} --| WHAT the respective properties of materials/substances/fluids are,
} --| HOW they have ( practically ) to be used safely,
} --| WHAT the consequences eventually will be (personal and for others) if
} --| handled unsafe, and
} --| HOW to dispose of (remnants, by-products of reactions, etc.) properly.
} --|
} --| (as an example for awareness: read:
} --| http://www.hull.ac.uk/chemstores/coshhadv.html (COSHH- Control of
} --| Substances Hazardous to Health, Univ.Hull, UK:  'part of the U.K.'s CRIMI=
} NAL
} --| LAW' !)
} --|
} --| Anybody who has |--studied--| properties of chemicals etc., their
} handling an=
} d
} --| use properly and correctly will know how to proceed with the use of mater=
} ial
} --| and substances we use in an EM-lab. If this has not be done, no one shoul=
} d
} --| do --|things|-- he never got explained and does not know about
} consequences i=
} n
} --| case of misuse.
} --|
} --| There IMO is no need to disallow or eventually prohibit (necessary) actio=
} ns
} --| which to the unexperienced (perhaps) will be harmful, but to the experien=
} ced
} --| does not apply seriously.
} --| So, last but not least: it is the responsibilty of the "experienced" to
} --| teach the unexperienced... so you as the |--experienced--| decide
} what can be
} --| done/shown and what CANNOT be done /shown.
} --|
} --| Concerning: Ice-Cream-experiments with LN2, those perhaps are an
} --| "interesting" demonstration for a new practical application but easily
} --| can/could be safely substituted by |--old fashioned--| ice-cream-making (e.g
} --| lowering the temperature of the fluids by using eg. the  pot-freezer meth=
} od:
} --| the temperature of the ingredients is reduced by placing them inside a tu=
} b
} --| filled with ice and salt) Cf. http://en.wikipedia.org/wiki/Ice_cream .
} --|
} --| At the end: just only pointing to the long MSA thread on --|N2 gas-LN2|--
} --| starting May 2nd 2009
} --| Conc: [Microscopy] Nitrogen leak (JEOL 6701F SEM)
} --|
} --|
} --| Regards,
} --| Wolfgang MUSS
} --| Head EM-Lab, Pathology
} --| SALK-LKH (Gen. Hosp.)
} --| SALZBURG Austria
} --|
} --|
} --|
} --|
} --| --| -----Urspr=C3=83=C6=92=C3=82=C2=BCngliche Nachricht-----
} --| --| Von: richard.ross-at-allisontransmission.com
} --| --| [mailto:richard.ross-at-allisontransmission.com]
} --| --| Gesendet: Dienstag, 26. J=C3=83=C6=92=C3=82=C2=A4nner 2010 14:57
} --| --| An: Mu=C3=83=C6=92=C3=85=C2=B8 Wolfgang
} --| --| Betreff: [Microscopy] Re: Liquid Nitrogen Safety [Ice-Cream made with
} --| --| LN2, concerns about]
} --| --|
} --| --|
} --| -------------------------------------------------------------------------=
} -
} --| --| The Microscopy ListServer -- CoSponsor:  The Microscopy Society of
} --| America
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} ---
} --| --| Way back when, Chuck Fiori used to put on a demo with LN2 at the LeHigh
} --| SEM school.
} --| Chuck wanted to show that sometimes its more hazardous to trap LN2 agains=
} t
} --| the body with protective gear than to give the liedenfrost effect room to
} --| create its protective blanket of gas. If the LN2 was provided, he would
} --| eventually take some into his mouth and blow 'smoke' rings. He told of a
} --| time when he accidentally swallowed some of the LN2 and the resulting
} --| stomach gas pressure caused him to black out, falling to the stage
} --| unconscious.
} --| Ultimately the excess pressure relieved itself as a belch and Chuck came
} --| to.
} --| Chuck certainly taught me things in ways that I remember! |--
} --| --|
} --| --| =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
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} --| 27, 37 -- Subject: [Microscopy] Re: Liquid Nitrogen Safety
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} --| 41, 25 -- Subject: FW: [Microscopy]  Re: Liquid Nitrogen Safety
} --| 41, 25 -- Date: Tue, 26 Jan 2010 16:30:53 -0600
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} --| 41, 25 -- From: "Gibbon, Donald L." |--donald.gibbon-at-matcoinc.com--|
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} --| microscopy-at-microscopy.com--|
} --| 55, 28 -- Subject: RE: [Microscopy] FW:  Re: Liquid Nitrogen Safety
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} "America believes in education; the average professor earns more money in a
} year than a professional athlete earns in a whole week." Evan Esar
}
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}
} I prefer the hand-cranked version, myself. =C2=A0Not only does it give more=
} direct tactile feedback on the overall thickness of the ice cream, but it =
} gives the baby (To me) cousins something to do that is out of the way of my=
} kitchen chaos on Thanksgiving!|--div--|
} |--br--||--/div--||--div--|As for the health aspect, there was a study
} that I read with=
} in the last three or four years (I can't remember anything specific) th=
} at said that the largest contributor to weight gain when eating is the ment=
} al state of the eater. =C2=A0I was discussing this with a colleague of mine=
} and we came to the conclusion that it must be true, based on the French. =
} =C2=A0(My apologies at this generalization, but I am basing my experience o=
} n the four incredible weeks I spent wandering around the French countryside=
} ...) =C2=A0|--/div--|
} |--div--||--br--||--/div--||--div--|The French eat wonderfully
} decadent, flavorful, and ric=
} h foods, and drink gallons of truly elegant wine. =C2=A0Neither me nor my c=
} olleague remembered seeing any obese French people (We're sure they exi=
} st, but they don't seem to be the norm as it seems in the US). =C2=A0We=
} also experienced the wonderful French culture of taking your time while ea=
} ting, enjoying both the meal and the company, not rushing through it.|--/div--|
} |--div--||--br--||--/div--||--div--|We could also come up with more
} French nationals who li=
} ved to respectable old age than we could of any other
} nationality.|--/div--||--di=
} v--||--br--||--/div--||--div--|I know that this is in no way a
} scientific observation, bu=
} t it's still enough to make me want to wander around rural France for t=
} he rest of my life eating hand-churned ice cream...|--/div--|
} |--div--||--br--||--/div--||--div--|Great. =C2=A0Now I'm hungry.
} =C2=A0First time this =
} list has done that to
} me...|--/div--||--div--||--br--||--/div--||--div--|--Justin A.
} Kraft|--/div=
} --||--div--|Professional Lab Rat/High Energy Physics Server
} Monkey|--/div--||--div--|Flor=
} ida International University|--/div--|
} |--div--|Miami, FL|--br--||--br--||--div class=3D"gmail_quote"--|On
} Tue, Jan 26, 2010 at 6:=
} 19 PM,  |--span dir=3D"ltr"--| {|--a
} href=3D"mailto:kenconverse-at-qualityimages.b=
} iz"--|kenconverse-at-qualityimages.biz|--/a--|} |--/span--|
} wrote:|--br--||--blockquote clas=
} s=3D"gmail_quote" style=3D"margin:0 0 0 .8ex;border-left:1px #ccc solid;pad=
} ding-left:1ex;"--|
} |--div class=3D"im"--||--br--|
} |--br--|
} |--br--|
} ---------------------------------------------------------------------------=
} -|--br--|
} The Microscopy ListServer -- CoSponsor: =C2=A0The Microscopy Society of Ame=
} rica|--br--|
} To =C2=A0Subscribe/Unsubscribe -- |--a href=3D"http://www.microscopy.com/Micr=
} oscopyListserver
} On-Line" target=3D"_blank"--|http://www.microscopy.com/MicroscopyListserver|--b=
} r--|
} On-Line|--/a--| Help |--a
} href=3D"http://www.microscopy.com/MicroscopyListserver/=
} FAQ.html" target=3D"_blank"--|http://www.microscopy.com/MicroscopyListserver/=
} FAQ.html|--/a--||--br--|
} ---------------------------------------------------------------------------=
} -|--br--|
} |--br--|
} |--/div--|Don,|--br--|
} You're right! =C2=A0It's a dirty job, but somebody's got to do =
} it. =C2=A0My wife and I got an electric ice cream maker as a wedding gift t=
} hat we haven't used nearly enough. =C2=A0My favorite recipe not only in=
} cludes the heavy cream, but a half dozen eggs. =C2=A0To die for...|--br--|
}
} |--br--|
} Of course the kill-joys will tell us that the heavy cream, the cholesterol =
} in the eggs and all the sugar will kill us, not to mention the environmenta=
} l effects of all the cattle and chickens, plus the inhumane conditions they=
} are kept in.|--br--|
}
} |--br--|
} I'll have another serving of that custard ice cream recipe, thank you.|--=
} br--|
} |--div class=3D"im"--||--br--|
} Ken Converse|--br--|
} owner|--br--|
} |--br--|
} QUALITY IMAGES|--br--|
} Servicing Scanning Electron Microscopes|--br--|
} Since 1981|--br--|
} 474 So. Bridgton Rd.|--br--|
} Bridgton, ME =C2=A004009|--br--|
} 207-647-4348|--br--|
} Fax 207-647-2688|--br--|
} |--a href=3D"mailto:kenconverse-at-qualityimages.biz"--|kenconverse-at-qualityimages.=
} biz|--/a--||--br--|
} |--a href=3D"http://qualityimages.biz"
} target=3D"_blank"--|qualityimages.biz|--/a=
} --||--br--|
} |--br--|
} |--br--|
} -----Original Message-----|--br--|
} |--/div--||--div class=3D"im"--|X-from: |--a
} href=3D"mailto:donald.gibbon-at-matcoinc.co=
} m"--|donald.gibbon-at-matcoinc.com|--/a--| [mailto:|--a
} href=3D"mailto:donald.gibbon-at-m=
} atcoinc.com"--|donald.gibbon-at-matcoinc.com|--/a--|]|--br--|
} Sent: Tuesday, January 26, 2010 5:34 PM|--br--|
} To: |--a href=3D"mailto:kenconverse-at-qualityimages.biz"--|kenconverse-at-qualityima=
} ges.biz|--/a--||--br--|
} |--/div--||--div class=3D"im"--|Subject: [Microscopy] FW: Re: Liquid
} Nitrogen Safet=
} y|--br--|
} |--br--|
} |--br--|
} |--br--|
} |--br--|
} ---------------------------------------------------------------------------=
} -|--br--|
} The Microscopy ListServer -- CoSponsor: =C2=A0The Microscopy Society of Ame=
} rica|--br--|
} To =C2=A0Subscribe/Unsubscribe -- |--a href=3D"http://www.microscopy.com/Micr=
} oscopyListserver
} On-Line" target=3D"_blank"--|http://www.microscopy.com/MicroscopyListserver|--b=
} r--|
} On-Line|--/a--| Help |--a
} href=3D"http://www.microscopy.com/MicroscopyListserver/=
} FAQ.html" target=3D"_blank"--|http://www.microscopy.com/MicroscopyListserver/=
} FAQ.html|--/a--||--br--|
} ---------------------------------------------------------------------------=
} -|--br--|
} |--br--|
} |--br--|
} |--br--|
} |--br--|
} |--/div--|As a semi-professional ice cream maker, I'd like to second Wolfga=
} ng Muss's reference to the old-fashioned but utterly reliable and safe =
} method of using a hand-cranked ice cream freezer rather than any other opti=
} on. There are reasons why this works well, which, if ignored, explain why i=
} t doesn't work sometimes! Making ice cream is an exercise in heat trans=
} fer and crystal growth. The heat transfer surface is the metal wall of the =
} tub which is rotated by the crank. Counter-rotating inside the tube is a pa=
} ir of wooden paddles, keeping that surface clean and free of ice, constantl=
} y removing the freezing cream. The idea is to get a eutectic mixture of sal=
} t and ice on the outside, i.e., minimum possible temperature in this system=
} , dump in the cream and start cranking. Don't ever stop for even one se=
} cond. i.e., keep that surface clean, and in six-eight minutes your job will=
} be done. The idea is to maximize the number of crystals and minimize their=
} size. It is NOT a tedious process, !|--br--|
}
} =C2=A0hardly enough to get winded.|--br--|
} |--br--|
} My most sought-after publication in my entire life was my paper on The Ther=
} modynamics of Homemade Ice Cream, published in the Journal Of Chemical Educ=
} ation. I still have the thermocouple-fitted ice cream freezer in which we m=
} ade many gallons of ice cream, doing the experimental work for this paper. =
} Somebody had to do it! And sticking your finger in the ice cream will not h=
} arm you at all!|--br--|
}
} |--br--|
} Donald L. Gibbon|--br--|
} |--div class=3D"im"--||--br--|
} -----Original Message-----|--br--|
} X-from: |--a href=3D"mailto:W.Muss-at-salk.at"--|W.Muss-at-salk.at|--/a--|
} [mailto:|--a hre=
} f=3D"mailto:W.Muss-at-salk.at"--|W.Muss-at-salk.at|--/a--|]|--br--|
} Sent: Tuesday, January 26, 2010 12:23 PM|--br--|
} To: Gibbon, Donald L.|--br--|
} Subject: [Microscopy] Re: Liquid Nitrogen Safety|--br--|
} |--br--|
} |--br--|
} |--br--|
} |--br--|
} ---------------------------------------------------------------------------=
} -|--br--|
} The Microscopy ListServer -- CoSponsor: =C2=A0The Microscopy Society of Ame=
} rica|--br--|
} To =C2=A0Subscribe/Unsubscribe -- |--a href=3D"http://www.microscopy.com/Micr=
} oscopyListserver
} On-Line" target=3D"_blank"--|http://www.microscopy.com/MicroscopyListserver|--b=
} r--|
} On-Line|--/a--| Help |--a
} href=3D"http://www.microscopy.com/MicroscopyListserver/=
} FAQ.html" target=3D"_blank"--|http://www.microscopy.com/MicroscopyListserver/=
} FAQ.html|--/a--||--br--|
} ---------------------------------------------------------------------------=
} -|--br--|
} |--br--|
} |--/div--|Good morning, good afternoon, good evening - hopefully any somewhere =
} will apply,|--br--|
} hello, dear colleagues,|--br--|
} |--br--|
} Just to add my 2 =C3=83=C2=A2=C3=A2=E2=82=AC=C5=A1=C3=82=C2=AC-cents (apolo=
} gize if the message became too long):|--br--|
} |--br--|
} Yes, we can discuss about children STICK THEIR FINGERS IN THE LIQUID NITROG=
} EN and other stuff.... =C2=A0 in general,|--br--|
} since in EM we sometimes have to work with 'cruel' things and subst=
} ances (and others a little bit more often).|--br--|
} Cryogenics also CAN be hazardous (as } hot { water is/can be), dependin=
} g on how we are } working with { it.|--br--|
} If we couldn't do that safe(ly) it necessarily / perhaps would have bee=
} n forbidden for a long time.|--br--|
} |--br--|
} There are a lot of sources on the web one can find concerning use and misus=
} e of e. g. liquid nitrogen:|--br--|
} for example:|--br--|
} |--br--|
} |--a href=3D"http://www.altair.org/hazard.html" target=3D"_blank"--|http://www.=
} altair.org/hazard.html|--/a--| =C2=A0(DONT DIE ! Laboratory Hazards: Safety, Pr=
} evention, First Aid, C 2001-2010)|--br--|
} |--br--|
} |--a href=3D"http://www.reachoutmichigan.org/funexperiments/agesubject/lesson=
} s/nitrogen.html" target=3D"_blank"--|http://www.reachoutmichigan.org/funexper=
} iments/agesubject/lessons/nitrogen.html|--/a--||--br--|
} ( "Fun experiments" website last updated 2003)|--br--|
} |--br--|
} |--a href=3D"http://smb.slac.stanford.edu/users_guide/manual/Experiment_polic=
} ies.html#SECTION00027000000000000000" target=3D"_blank"--|http://smb.slac.sta=
} nford.edu/users_guide/manual/Experiment_policies.html#SECTION00027000000000=
} 000000|--/a--||--br--|
}
} (describes hazards and proper handling procedures for work with liquid nitr=
} ogen, 2010)|--br--|
} |--br--|
} |--a href=3D"http://stores.biochem.uiowa.edu/Pages/ln2msds.htm" target=3D"_bl=
} ank"--|http://stores.biochem.uiowa.edu/Pages/ln2msds.htm|--/a--||--br--|
} ("old" but IMO "good" example for a US-Univ's Safet=
} y Data Sheet)|--br--|
} |--br--|
} |--a href=3D"http://www.chaosscience.org.uk/dem/public_html//article.php?stor=
} y=3D20031216175107931" target=3D"_blank"--|http://www.chaosscience.org.uk/dem=
} /public_html//article.php?story=3D20031216175107931|--/a--||--br--|
} (Liquid Nitrogen Risk Assessment, 16/12/03 )|--br--|
} |--br--|
} Also, some Labs are questioning in advance for safety issues on the things =
} someone will bring into the Lab:|--br--|
} e.g. |--a href=3D"http://www.sbc.anl.gov/pdfs/hazard_assessment_form.pdf" tar=
} get=3D"_blank"--|http://www.sbc.anl.gov/pdfs/hazard_assessment_form.pdf|--/a--|
} =
} =C2=A0(take a look for 'cryogens', Argonne Natl. Lab, SBC Hazard As=
} sessment Form), perhaps just to be on the safe side for their own staff peo=
} ple and Lab's interior.|--br--|
}
} |--br--|
} AND finally, YES, LN2-ingestion - to take a mouthfull of LN2 and blast it o=
} ff - only for the EXPERIENCED CAN BE a junky-funky trick, BUT -|--br--|
} on the other side - for the unexperienced (unfortunately also for the '=
} careless' experienced I have to admit!) - it could be not only dangerou=
} s but LIFE threatening: =C2=A0cf.|--br--|
} |--a href=3D"http://pediatrics.aappublications.org/cgi/reprint/105/1/121" tar=
} get=3D"_blank"--|http://pediatrics.aappublications.org/cgi/reprint/105/1/121|--=
} /a--||--br--|
} ( Benjamin Z. Koplewitz, et al.... =C2=A0Gastric Perforation Attributable t=
} o Liquid Nitrogen Ingestion,|--br--|
} in: =C2=A0Pediatrics 2000;105;121-123, open Access)|--br--|
} |--br--|
} A similar situation has been reported on |--a href=3D"http://www.darwinawards=
} .com/personal/personal2000-25.html" target=3D"_blank"--|http://www.darwinawar=
} ds.com/personal/personal2000-25.html|--/a--|,|--br--|
} which eventually was } awarded { with the DARWIN AWARD in 2000 (=3D=3D&=
} gt;cit: "The Darwin Awards salute the improvement of the human genome =
} by honoring those who accidentally remove themselves from it..." end o=
} f cit.)|--br--|
}
} |--br--|
} I personally have no doubt that it IS possible (and I have done that for de=
} monstration of "Leidenfrost phenomenon" a lot of times way back..=
} .) to place one or two fingers, eventually(and forsure) the whole hand into=
} liquid nitrogen for 1-2 seconds, PROVIDED some necessary PREREQUISITES whi=
} ch - IN ADVANCE - have to be theoretically explained and are to be met to/f=
} or those } trying { such an "adventure" and IMO only should b=
} e done in a SUPERVISED situation.|--br--|
}
} |--br--|
} Despite (or better: BECAUSE) respecting all those "little" {pr=
} oblem-makers} in our daily work|--br--|
} (if this is not done by other staff members, I have to to do such work lite=
} rally by myself) I am not in a blue funk of perhaps desastrous results of u=
} sing/handling all those substances like|--br--|
} OsO4 (hazard, toxic), p-phenylenediamine (pesticide, toxic), uranyl-acetate=
} from stock powder (radioactive), cryogens (like LN2, precooled isobutane, =
} also Freon gas [some residual inventory] if the pressurized can will be use=
} d inverted) and other substances/chemicals (mostly used in very small quant=
} ities), not to forget all those "may be hazardous resins" out in =
} the dark...|--br--|
}
} |--br--|
} WHY I am NOT AFRAID/do not fear those: =C2=A0since I have learned - before =
} handling and using those -|--br--|
} WHAT the respective properties of materials/substances/fluids are,|--br--|
} HOW they have ( practically ) to be used safely,|--br--|
} WHAT the consequences eventually will be (personal and for others) if handl=
} ed unsafe, and|--br--|
} HOW to dispose of (remnants, by-products of reactions, etc.) properly.|--br--|
} |--br--|
} (as an example for awareness: read: =C2=A0|--a href=3D"http://www.hull.ac.uk/=
} chemstores/coshhadv.html" target=3D"_blank"--|http://www.hull.ac.uk/chemstore=
} s/coshhadv.html|--/a--| (COSHH- Control of Substances Hazardous to Health, Univ=
} .Hull, UK: =C2=A0'part of the U.K.'s CRIMINAL LAW' !)|--br--|
}
} |--br--|
} Anybody who has {studied} properties of chemicals etc., their handlin=
} g and use properly and correctly will know how to proceed with the use of m=
} aterial and substances we use in an EM-lab. If this has not be done, no one=
} should do } things { he never got explained and does not know about co=
} nsequences in case of misuse.|--br--|
}
} |--br--|
} There IMO is no need to disallow or eventually prohibit (necessary) actions=
} which to the unexperienced (perhaps) will be harmful, but to the experienc=
} ed does not apply seriously.|--br--|
} So, last but not least: it is the responsibilty of the "experienced&qu=
} ot; to teach the unexperienced... so you as the {experienced} decide =
} what can be done/shown and what CANNOT be done /shown.|--br--|
} |--br--|
} Concerning: Ice-Cream-experiments with LN2, those perhaps are an "inte=
} resting" demonstration for a new practical application but easily can/=
} could be safely substituted by {old fashioned} ice-cream-making (e.g =
} lowering the temperature of the fluids by using eg. the =C2=A0pot-freezer m=
} ethod: the temperature of the ingredients is reduced by placing them inside=
} a tub filled with ice and salt) Cf. |--a href=3D"http://en.wikipedia.org/wik=
} i/Ice_cream" target=3D"_blank"--|http://en.wikipedia.org/wiki/Ice_cream|--/a--|
} .=
} |--br--|
}
} |--br--|
} At the end: just only pointing to the long MSA thread on } N2 gas-LN2 {=
} =C2=A0 starting May 2nd 2009|--br--|
} Conc: [Microscopy] Nitrogen leak (JEOL 6701F SEM)|--br--|
} |--br--|
} |--br--|
} Regards,|--br--|
} Wolfgang MUSS|--br--|
} Head EM-Lab, Pathology|--br--|
} SALK-LKH (Gen. Hosp.)|--br--|
} SALZBURG Austria|--br--|
} |--br--|
} |--br--|
} |--br--|
} |--br--|
} } -----Urspr=C3=83=C6=92=C3=82=C2=BCngliche Nachricht-----|--br--|
} } Von: |--a href=3D"mailto:richard.ross-at-allisontransmission.com"--|richard.r=
} oss-at-allisontransmission.com|--/a--||--br--|
} } [mailto:|--a href=3D"mailto:richard.ross-at-allisontransmission.com"--|richar=
} d.ross-at-allisontransmission.com|--/a--|]|--br--|
} } Gesendet: Dienstag, 26. J=C3=83=C6=92=C3=82=C2=A4nner 2010 14:57|--br--|
} } An: Mu=C3=83=C6=92=C3=85=C2=B8 Wolfgang|--br--|
} } Betreff: [Microscopy] Re: Liquid Nitrogen Safety [Ice-Cream made with|--=
} br--|
} } LN2, concerns about]|--br--|
} |--div class=3D"im"--|} |--br--|
} } ----------------------------------------------------------------------=
} ----|--br--|
} } The Microscopy ListServer -- CoSponsor: =C2=A0The Microscopy Society o=
} f America|--br--|
} } To =C2=A0Subscribe/Unsubscribe -- |--a href=3D"http://www.microscopy.com=
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} } ----------------------------------------------------------------------=
} ------|--br--|
} |--/div--|} Way back when, Chuck Fiori used to put on a demo with LN2 at the=
} LeHigh SEM school.|--br--|
} Chuck wanted to show that sometimes its more hazardous to trap LN2 against =
} the body with protective gear than to give the liedenfrost effect room to c=
} reate its protective blanket of gas. If the LN2 was provided, he would even=
} tually take some into his mouth and blow 'smoke' rings. He told of =
} a time when he accidentally swallowed some of the LN2 and the resulting sto=
} mach gas pressure caused him to black out, falling to the stage unconscious=
} .|--br--|
}
} Ultimately the excess pressure relieved itself as a belch and Chuck came to=
} .|--br--|
} Chuck certainly taught me things in ways that I remember! {|--br--|
} } |--br--|
} } =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=
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} } 1, 25 -- From |--a href=3D"mailto:richard.ross-at-allisontransmission.com"--|=
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} } 1, 25 -- Date: Tue, 26 Jan 2010 08:52:56 -0500|--br--|
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--
"America believes in education; the average professor earns more money
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Esar


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From: eldesouky.ammar-at-ars.usda.gov
Date: Wed, 27 Jan 2010 08:48:31 -0600
Subject: [Microscopy] viaWWW: TEM prep training

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Email: eldesouky.ammar-at-ars.usda.gov
Name: Eldesouky Ammar

Organization: USDA-ARS

Title-Subject: [Filtered] TEM prep training

Message: Hi
A former student of mine, now an Assoc. Prof. at Cairo Univ. wants to
attend a workshop on TEM-use and sample preparations in the USA or
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future. Thanks
E. Ammar

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From: PWebster-at-hei.org
Date: Wed, 27 Jan 2010 09:25:11 -0600
Subject: [Microscopy] viaWWW: TEM prep training

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Tell your student to check out the EMBO web site for a course that will take place in Oslo, Norway this year (run by Norbert Roos). The course covers basic specimen preparation, use of resins, cryosectioning, immunocytochemistry and the application of stereological methods. It is unique in many respects - students can bring their own specimens and for most participants the course is free. However, acceptance onto the course is competitive.

Regards,

Paul Webster
House Ear Institute
2100 W 3rd St
Los Angeles
CA 90057



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Email: eldesouky.ammar-at-ars.usda.gov
Name: Eldesouky Ammar

Organization: USDA-ARS

Title-Subject: [Filtered] TEM prep training

Message: Hi
A former student of mine, now an Assoc. Prof. at Cairo Univ. wants to
attend a workshop on TEM-use and sample preparations in the USA or
Europe. Does anyone have info about such a workshop in the near
future. Thanks
E. Ammar

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From: randerson20-at-tampabay.rr.com
Date: Wed, 27 Jan 2010 09:25:47 -0600
Subject: [Microscopy] FW: Re: Liquid Nitrogen Safety

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Ken,

You are right about the killjoys, but did you ever consider that:

1. The Japanese eat very little fat and suffer fewer heart attacks than
the English and Americans.
2. The mexicans eat a lot of fat and suffer fewer heart attacks than the
English and Americans.
3. The Chinese drink very little red wine and suffer fewer heart attacks
than the English and Americans.
4. The Italians drik a lot of red wine and suffer fewer heart attacks
than the English and Americans.
CONCLUSION
Eat and drink what you like. Speaking English is apparently what kills you!*

*stolen from Anaspec Info, Issue 59/09, Fall 2009, Gauteng, South Africa.

Ron Anderson

kenconverse-at-qualityimages.biz wrote:
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} Don,
} You're right! It's a dirty job, but somebody's got to do it. My wife and I got an electric ice cream maker as a wedding gift that we haven't used nearly enough. My favorite recipe not only includes the heavy cream, but a half dozen eggs. To die for...
}
} Of course the kill-joys will tell us that the heavy cream, the cholesterol in the eggs and all the sugar will kill us, not to mention the environmental effects of all the cattle and chickens, plus the inhumane conditions they are kept in.
}
} I'll have another serving of that custard ice cream recipe, thank you.
}
} Ken Converse
} owner
}
} QUALITY IMAGES
} Servicing Scanning Electron Microscopes
} Since 1981
} 474 So. Bridgton Rd.
} Bridgton, ME 04009
} 207-647-4348
} Fax 207-647-2688
} kenconverse-at-qualityimages.biz
} qualityimages.biz
}
}
} -----Original Message-----
} X-from: donald.gibbon-at-matcoinc.com [mailto:donald.gibbon-at-matcoinc.com]
} Sent: Tuesday, January 26, 2010 5:34 PM
} To: kenconverse-at-qualityimages.biz
} Subject: [Microscopy] FW: Re: Liquid Nitrogen Safety
}
}
}
}
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}
}
} As a semi-professional ice cream maker, I'd like to second Wolfgang Muss's reference to the old-fashioned but utterly reliable and safe method of using a hand-cranked ice cream freezer rather than any other option. There are reasons why this works well, which, if ignored, explain why it doesn't work sometimes! Making ice cream is an exercise in heat transfer and crystal growth. The heat transfer surface is the metal wall of the tub which is rotated by the crank. Counter-rotating inside the tube is a pair of wooden paddles, keeping that surface clean and free of ice, constantly removing the freezing cream. The idea is to get a eutectic mixture of salt and ice on the outside, i.e., minimum possible temperature in this system, dump in the cream and start cranking. Don't ever stop for even one second. i.e., keep that surface clean, and in six-eight minutes your job will be done. The idea is to maximize the number of crystals and minimize their size. It is NOT a tedious process, !
} hardly enough to get winded.
}
} My most sought-after publication in my entire life was my paper on The Thermodynamics of Homemade Ice Cream, published in the Journal Of Chemical Education. I still have the thermocouple-fitted ice cream freezer in which we made many gallons of ice cream, doing the experimental work for this paper. Somebody had to do it! And sticking your finger in the ice cream will not harm you at all!
}
} Donald L. Gibbon
}
} -----Original Message-----
} X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at]
} Sent: Tuesday, January 26, 2010 12:23 PM
} To: Gibbon, Donald L.
} Subject: [Microscopy] Re: Liquid Nitrogen Safety
}
}
}
}
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} Good morning, good afternoon, good evening - hopefully any somewhere will apply,
} hello, dear colleagues,
}
} Just to add my 2 €-cents (apologize if the message became too long):
}
} Yes, we can discuss about children STICK THEIR FINGERS IN THE LIQUID NITROGEN and other stuff.... in general,
} since in EM we sometimes have to work with 'cruel' things and substances (and others a little bit more often).
} Cryogenics also CAN be hazardous (as } hot { water is/can be), depending on how we are } working with { it.
} If we couldn't do that safe(ly) it necessarily / perhaps would have been forbidden for a long time.
}
} There are a lot of sources on the web one can find concerning use and misuse of e. g. liquid nitrogen:
} for example:
}
} http://www.altair.org/hazard.html (DONT DIE ! Laboratory Hazards: Safety, Prevention, First Aid, C 2001-2010)
}
} http://www.reachoutmichigan.org/funexperiments/agesubject/lessons/nitrogen.html
} ( "Fun experiments" website last updated 2003)
}
} http://smb.slac.stanford.edu/users_guide/manual/Experiment_policies.html#SECTION00027000000000000000
} (describes hazards and proper handling procedures for work with liquid nitrogen, 2010)
}
} http://stores.biochem.uiowa.edu/Pages/ln2msds.htm
} ("old" but IMO "good" example for a US-Univ's Safety Data Sheet)
}
} http://www.chaosscience.org.uk/dem/public_html//article.php?story=20031216175107931
} (Liquid Nitrogen Risk Assessment, 16/12/03 )
}
} Also, some Labs are questioning in advance for safety issues on the things someone will bring into the Lab:
} e.g. http://www.sbc.anl.gov/pdfs/hazard_assessment_form.pdf (take a look for 'cryogens', Argonne Natl. Lab, SBC Hazard Assessment Form), perhaps just to be on the safe side for their own staff people and Lab's interior.
}
} AND finally, YES, LN2-ingestion - to take a mouthfull of LN2 and blast it off - only for the EXPERIENCED CAN BE a junky-funky trick, BUT -
} on the other side - for the unexperienced (unfortunately also for the 'careless' experienced I have to admit!) - it could be not only dangerous but LIFE threatening: cf.
} http://pediatrics.aappublications.org/cgi/reprint/105/1/121
} ( Benjamin Z. Koplewitz, et al.... Gastric Perforation Attributable to Liquid Nitrogen Ingestion,
} in: Pediatrics 2000;105;121-123, open Access)
}
} A similar situation has been reported on http://www.darwinawards.com/personal/personal2000-25.html,
} which eventually was } awarded { with the DARWIN AWARD in 2000 (==} cit: "The Darwin Awards salute the improvement of the human genome by honoring those who accidentally remove themselves from it..." end of cit.)
}
} I personally have no doubt that it IS possible (and I have done that for demonstration of "Leidenfrost phenomenon" a lot of times way back...) to place one or two fingers, eventually(and forsure) the whole hand into liquid nitrogen for 1-2 seconds, PROVIDED some necessary PREREQUISITES which - IN ADVANCE - have to be theoretically explained and are to be met to/for those } trying { such an "adventure" and IMO only should be done in a SUPERVISED situation.
}
} Despite (or better: BECAUSE) respecting all those "little" {problem-makers} in our daily work
} (if this is not done by other staff members, I have to to do such work literally by myself) I am not in a blue funk of perhaps desastrous results of using/handling all those substances like
} OsO4 (hazard, toxic), p-phenylenediamine (pesticide, toxic), uranyl-acetate from stock powder (radioactive), cryogens (like LN2, precooled isobutane, also Freon gas [some residual inventory] if the pressurized can will be used inverted) and other substances/chemicals (mostly used in very small quantities), not to forget all those "may be hazardous resins" out in the dark...
}
} WHY I am NOT AFRAID/do not fear those: since I have learned - before handling and using those -
} WHAT the respective properties of materials/substances/fluids are,
} HOW they have ( practically ) to be used safely,
} WHAT the consequences eventually will be (personal and for others) if handled unsafe, and
} HOW to dispose of (remnants, by-products of reactions, etc.) properly.
}
} (as an example for awareness: read: http://www.hull.ac.uk/chemstores/coshhadv.html (COSHH- Control of Substances Hazardous to Health, Univ.Hull, UK: 'part of the U.K.'s CRIMINAL LAW' !)
}
} Anybody who has {studied} properties of chemicals etc., their handling and use properly and correctly will know how to proceed with the use of material and substances we use in an EM-lab. If this has not be done, no one should do } things { he never got explained and does not know about consequences in case of misuse.
}
} There IMO is no need to disallow or eventually prohibit (necessary) actions which to the unexperienced (perhaps) will be harmful, but to the experienced does not apply seriously.
} So, last but not least: it is the responsibilty of the "experienced" to teach the unexperienced... so you as the {experienced} decide what can be done/shown and what CANNOT be done /shown.
}
} Concerning: Ice-Cream-experiments with LN2, those perhaps are an "interesting" demonstration for a new practical application but easily can/could be safely substituted by {old fashioned} ice-cream-making (e.g lowering the temperature of the fluids by using eg. the pot-freezer method: the temperature of the ingredients is reduced by placing them inside a tub filled with ice and salt) Cf. http://en.wikipedia.org/wiki/Ice_cream .
}
} At the end: just only pointing to the long MSA thread on } N2 gas-LN2 { starting May 2nd 2009
} Conc: [Microscopy] Nitrogen leak (JEOL 6701F SEM)
}
}
} Regards,
} Wolfgang MUSS
} Head EM-Lab, Pathology
} SALK-LKH (Gen. Hosp.)
} SALZBURG Austria
}
}
}
}
}
} } -----Ursprüngliche Nachricht-----
} } Von: richard.ross-at-allisontransmission.com
} } [mailto:richard.ross-at-allisontransmission.com]
} } Gesendet: Dienstag, 26. Jänner 2010 14:57
} } An: Muß Wolfgang
} } Betreff: [Microscopy] Re: Liquid Nitrogen Safety [Ice-Cream made with
} } LN2, concerns about]
} }
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} } Way back when, Chuck Fiori used to put on a demo with LN2 at the LeHigh SEM school.
} }
} Chuck wanted to show that sometimes its more hazardous to trap LN2 against the body with protective gear than to give the liedenfrost effect room to create its protective blanket of gas. If the LN2 was provided, he would eventually take some into his mouth and blow 'smoke' rings. He told of a time when he accidentally swallowed some of the LN2 and the resulting stomach gas pressure caused him to black out, falling to the stage unconscious.
} Ultimately the excess pressure relieved itself as a belch and Chuck came to.
} Chuck certainly taught me things in ways that I remember! {
}
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From: paul_hazelton-at-umanitoba.ca
Date: Wed, 27 Jan 2010 09:27:32 -0600
Subject: [Microscopy] Re: Liquid Nitrogen Safety

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

Justin

Escoffier said that a recipe is only a list of ideas. So no, science
has not impacted on my cooking. I still have some great successes in
the kitchen, and some truly magnificent failures. However, science sure
has had an impact on my wine making, and for the better.

paul

--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 William Avenue
Winnipeg, Manitoba, Canada, R3E 0J9
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926



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From: lamiller-at-illinois.edu
Date: Wed, 27 Jan 2010 09:55:57 -0600
Subject: [Microscopy] Liquid Nitrogen Safety

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

chuckle...

Recipes.... who needs recipes?

That being said, I think science has had a lot of influence on my
cooking, and I sometimes long for lab ware to get things done!
I also consider what gets mixed when, what is soluble in fat etc.

However I was cooking long before science, so maybe it's the other way
around, how does my cooking affect my science?


Lou Ann



On Jan 27, 2010, at 9:36 AM, paul_hazelton-at-umanitoba.ca wrote:

}
}
}
} ----------------------------------------------------------------------------
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} America
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} ----------------------------------------------------------------------------
}
} Justin
}
} Escoffier said that a recipe is only a list of ideas. So no, science
} has not impacted on my cooking. I still have some great successes in
} the kitchen, and some truly magnificent failures. However, science
} sure
} has had an impact on my wine making, and for the better.
}
} paul
}
} --
} Paul R. Hazelton, PhD
} Viral Gastroenteritis Study Group
} University of Manitoba
} Department of Medical Microbiology
} 511 Basic Medical Sciences Building
} 745 William Avenue
} Winnipeg, Manitoba, Canada, R3E 0J9
} e-mail: paul_hazelton-at-umanitoba.ca
} paulhazelton-at-mts.net
} Phone: 204-789-3313 (w);
} 204-489-6924 (h)
} Cell: 204-781-6982
} Fax: 204-789-3926
}
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}
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{ { { { { { { { {} } } } } } } } } } } } } }
Lou Ann Miller, Service Supervisor
Center for Microscopic Imaging
College of Veterinary Medicine
University of Illinois MC=002

Room 1204 VMBSBld
2001 S Lincoln Ave
Urbana, IL 61821

217-244-1567
http://treefrog.cvm.uiuc.edu






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From: Chris.Guerin-at-dmbr.vib-UGent.be
Date: Wed, 27 Jan 2010 09:56:57 -0600
Subject: [Microscopy] Science and cooking

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

Hi All:

At the risk of wasting time with off topic posts I have to chime in.

Having been both a professional chef and a scientist I think I can say with some authority that if you are good at one chances are you will be good at the other, at least in as much as wet lab work goes. The combination of being able to follow a protocol (recipe) while at the same time having enough creativity and knowledge of the principles to experiment a bit and perhaps improve upon it are very advantageous in both the lab and the kitchen. Add to that some manual dexterity, patience and a sense of genuine joy in the possibility of achieving something great and you will get either a very good cook (and) or a very good scientist. Whenever I interview someone for a technical post I ask if they like to cook, those that do generally work out well in the lab.

I will also share that my first kitchen job was as a prep cook. For those who don't know a prep cook is the sorry creature who gets to do all the messy, difficult and downright disgusting jobs the chef no longer wants to do; so you see, it was also excellent training for being a grad student.

Best wishes, Chris

Christopher Guérin, Ph.D.
Leader, Microscopy Core
Dept. for Molecular Biomedical Research
Flanders Institute of Biotechnology (VIB)
University of Ghent, Belgium

==============================Original Headers==============================
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From: reinhard.rachel-at-biologie.uni-regensburg.de
Date: Wed, 27 Jan 2010 10:19:08 -0600
Subject: [Microscopy] TEM prep training

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

as Paul Webster said, see:

EMBO Practical Course "Electron microscopy and stereology in cell biology"

http://cwp.embo.org/pc10-19/

best regards,
Reinhard


--

PD Dr. Reinhard Rachel
Universitaet Regensburg
Centre for EM - NWF III -
-at-Institute for Anatomy
Universitaetsstr. 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29



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From: FMonson-at-wcupa.edu
Date: Wed, 27 Jan 2010 11:06:49 -0600
Subject: [Microscopy] Liquid Nitrogen Safety

Contents Retrieved from Microscopy Listserver Archives
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http://chestjournal.chestpubs.org/content/118/4/1150.full

Dear Lou Ann,

If you are going to cook, science should come first.

Cheers and chuckles,

Fred Monson

CMIRT
West Chester Universrsity
http://cmirt.wcupa.edu
________________________________________
X-from: lamiller-at-illinois.edu [lamiller-at-illinois.edu]
Sent: Wednesday, January 27, 2010 11:06 AM
To: Monson, Frederick

chuckle...

Recipes.... who needs recipes?

That being said, I think science has had a lot of influence on my
cooking, and I sometimes long for lab ware to get things done!
I also consider what gets mixed when, what is soluble in fat etc.

However I was cooking long before science, so maybe it's the other way
around, how does my cooking affect my science?


Lou Ann



On Jan 27, 2010, at 9:36 AM, paul_hazelton-at-umanitoba.ca wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Justin
}
} Escoffier said that a recipe is only a list of ideas. So no, science
} has not impacted on my cooking. I still have some great successes in
} the kitchen, and some truly magnificent failures. However, science
} sure
} has had an impact on my wine making, and for the better.
}
} paul
}
} --
} Paul R. Hazelton, PhD
} Viral Gastroenteritis Study Group
} University of Manitoba
} Department of Medical Microbiology
} 511 Basic Medical Sciences Building
} 745 William Avenue
} Winnipeg, Manitoba, Canada, R3E 0J9
} e-mail: paul_hazelton-at-umanitoba.ca
} paulhazelton-at-mts.net
} Phone: 204-789-3313 (w);
} 204-489-6924 (h)
} Cell: 204-781-6982
} Fax: 204-789-3926
}
}
}
} ==============================Original
} Headers==============================
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{ { { { { { { { {} } } } } } } } } } } } } }
Lou Ann Miller, Service Supervisor
Center for Microscopic Imaging
College of Veterinary Medicine
University of Illinois MC=002

Room 1204 VMBSBld
2001 S Lincoln Ave
Urbana, IL 61821

217-244-1567
http://treefrog.cvm.uiuc.edu






==============================Original Headers==============================
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==============================Original Headers==============================
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From: johnf-at-geology.wisc.edu
Date: Wed, 27 Jan 2010 11:34:56 -0600
Subject: [Microscopy] Reminder: EBSD 2010 conference May 2010

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

Please consider attending yourself and please
pass this on to potentially interested students,
postdocs, researchers and professors.

50% of the 125 open seats have already been
filled, so waiting much longer may mean missing
out on this great meeting, if EBSD is your
interest.

What: Microbeam Analysis Society's EBSD 2010 Topical Conference
When: Monday May 24-Wednesday May 26 (also several vendor events Sunday May 23
Where: University of Wisconsin, Madison, WI
Who: everyone from raw novices to experienced practitioners
Why: a one day tutorial with hands-on labs for
beginners, and 2 days of invited and contributed
talks on the latest topics, including sample prep
for both geological material and metals

There is abundant funding for student
participation, and any student using EBSD or
contemplating using EBSD in their research should
plan to attend! The deadline for applications for
financial aid is March 1 (you must be registered
first, but submission of an abstract is not
required.)

Abstract submission deadline is March 1.

Invited/confirmed Speakers:

Carl Boehlert, Michigan State University -
"In-situ SEM/EBSD observations of structural
alloys during elevated-temperature deformation"
Roy Geiss, NIST-Boulder -
"Case Studies of Elastic Strain Measurement
on Crystalline Materials using EBSD"
Brad Hacker, Univ of California, Santa Barbara -
"Seismic anisotrophy in Earth's crust"
Elisabetta Mariani, Liverpool University -
"Recrystallization in geological materials:
from in-situ to time series experiments"
Steve Reddy, Curtain University, Australia -
"Integrating EBSD with high spatial
resolution geochemistry & some geological
applications"
Aimo Winkelmann, MPI für Mikrostrukturphysik , Halle, Germany -
"Depth sensitivity and energy resolution of EBSD"

Also, geologists may be interested in the
NSF-supported "Structural Geology and Tectonics
Forum" being held immediately proceeding the EBSD
conference. It will be May 20-22 at UW-Madison,
and to get further details contact Richard Becker
(rabecker2-at-wisc.edu).


--
========================================================
John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480
Cameron Electron Microprobe Lab lab: (608) 265-4798
Department of Geoscience fax: (608) 262-0693
University of Wisconsin home: (608) 274-2245
1215 West Dayton St. email: johnf-at-geology.wisc.edu
Madison, WI 53706 amateur radio: WA3BTA
Personal http://www.geology.wisc.edu/~johnf/
Probe lab http://www.geology.wisc.edu/~johnf/sx51.html
Probe Sign Up Calender:
http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi

"The first rule of all intelligent tinkering is
to save every cog and wheel." -- Aldo Leopold

"For a successful technology, reality must take
precedence over public relations, for Nature
cannot be fooled." -- Richard P. Feynman


==============================Original Headers==============================
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From: lamiller-at-illinois.edu
Date: Wed, 27 Jan 2010 11:41:54 -0600
Subject: [Microscopy] Liquid Nitrogen Safety

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

I love it!!

:-)


On Jan 27, 2010, at 11:13 AM, FMonson-at-wcupa.edu wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} http://chestjournal.chestpubs.org/content/118/4/1150.full
}
} Dear Lou Ann,
}
} If you are going to cook, science should come first.
}
} Cheers and chuckles,
}
} Fred Monson
}
} CMIRT
} West Chester Universrsity
} http://cmirt.wcupa.edu
} ________________________________________
} X-from: lamiller-at-illinois.edu [lamiller-at-illinois.edu]
} Sent: Wednesday, January 27, 2010 11:06 AM
} To: Monson, Frederick
} Subject: [Microscopy] Liquid Nitrogen Safety
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} chuckle...
}
} Recipes.... who needs recipes?
}
} That being said, I think science has had a lot of influence on my
} cooking, and I sometimes long for lab ware to get things done!
} I also consider what gets mixed when, what is soluble in fat etc.
}
} However I was cooking long before science, so maybe it's the other way
} around, how does my cooking affect my science?
}
}
} Lou Ann
}
}

{ { { { { { { { {} } } } } } } } } } } } } }
Lou Ann Miller, Service Supervisor
Center for Microscopic Imaging
College of Veterinary Medicine
University of Illinois MC=002

Room 1204 VMBSBld
2001 S Lincoln Ave
Urbana, IL 61821

217-244-1567
http://treefrog.cvm.uiuc.edu






==============================Original Headers==============================
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13, 18 -- From: Lou Ann Miller {lamiller-at-illinois.edu}
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From: jae5-at-lehigh.edu
Date: Wed, 27 Jan 2010 12:49:53 -0600
Subject: [Microscopy] Liquid nitrogen and obesity (again)

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

Liquid nitrogen.

There is one important point that I did not see in this thread. If I
pour liquid nitrogen across the palm of my hand it does no harm - as
long as it runs off quickly. The gas layer protects me.

However, if I pour liquid nitrogen onto the back of my hand I will
almost certainly get a burn. The difference is that I have hair on the
back of my hand and it only takes a very small drop of liquid nitrogen
to be held in the same place by the hairs, for a burn to result. The
nitrogen must not be allowed to sit in the same place for any length of
time.

On a completely separate point, related to ice cream. If you want to
know why the Japanese suffer fewer heart attacks etc, read The Spirit
Level. This is the most important book of 2009 and probably (in my
view) the most important book of the decade.

Alwyn Eades

The Spirit Level: Why Greater Equality Makes Societies Stronger
R. Wilkinson and K. Pickett
Bloomsbury Press, 2009 (Publisher in the UK: Allen Lane)

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


==============================Original Headers==============================
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From: jae5-at-lehigh.edu
Date: Wed, 27 Jan 2010 13:49:18 -0600
Subject: [Microscopy] TEM in Avatar

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


I know that some people collect sightings of electron microscopes in
movies. Am I the only person who was so bored by Avatar that I spotted
a TEM (at least one, it might even have been two - I will have to wait
for the cable release to be sure) in one of the scenes in the lab?

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


==============================Original Headers==============================
4, 21 -- From jae5-at-lehigh.edu Wed Jan 27 13:49:17 2010
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4, 21 -- From: Alwyn Eades {jae5-at-lehigh.edu}
4, 21 -- Organization: Lehigh University
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From: TindallR-at-missouri.edu
Date: Wed, 27 Jan 2010 13:51:47 -0600
Subject: [Microscopy] TEM in Avatar

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

I am pretty sure I saw it too. At least a very TEM-like column.


Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




-----Original Message-----
X-from: jae5-at-lehigh.edu [mailto:jae5-at-lehigh.edu]
Sent: Wednesday, January 27, 2010 1:50 PM
To: Tindall, Randy D.


I know that some people collect sightings of electron microscopes in
movies. Am I the only person who was so bored by Avatar that I spotted
a TEM (at least one, it might even have been two - I will have to wait
for the cable release to be sure) in one of the scenes in the lab?

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


==============================Original Headers==============================
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From: StevenLe-at-BaylorHealth.edu
Date: Wed, 27 Jan 2010 14:01:39 -0600
Subject: [Microscopy] TEM in Avatar

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

I saw them as well. I thought one looked very similar to the FEI Technai series, and the other I can't remember the name of, but it has the smoked plastic around the column.


Steve Lee
Chief Technologist
Electron Microscopy Laboratory
Baylor University Medical Center
3500 Gaston Ave
Dallas, TX 75246
( 214.820.3302
Ê 214.820.4110
( stevenle-at-baylorhealth.edu


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Wednesday, January 27, 2010 1:57 PM
To: Lee, Steven

I am pretty sure I saw it too. At least a very TEM-like column.


Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




-----Original Message-----
X-from: jae5-at-lehigh.edu [mailto:jae5-at-lehigh.edu]
Sent: Wednesday, January 27, 2010 1:50 PM
To: Tindall, Randy D.


I know that some people collect sightings of electron microscopes in
movies. Am I the only person who was so bored by Avatar that I spotted
a TEM (at least one, it might even have been two - I will have to wait
for the cable release to be sure) in one of the scenes in the lab?

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


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26, 30 -- From StevenLe-at-BaylorHealth.edu Wed Jan 27 14:01:39 2010
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From: george.theodossiou-at-st-annes.ox.ac.uk
Date: Wed, 27 Jan 2010 14:37:08 -0600
Subject: [Microscopy] TEM in Avatar

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

There is also an orange column TEM in the TV series Dexter. When he's in his little lab.
I too have been bored by TV and movies.

More TEM's in movies I say....actually how bout an old VG HB501. I know the one here has more character and versatility than some leading actors and actresses out there and it works for free. The microscopist on the other hand will need food, drink and board.

________________________________________
X-from: StevenLe-at-BaylorHealth.edu [StevenLe-at-BaylorHealth.edu]
Sent: 27 January 2010 20:08
To: George Theodossiou

I saw them as well. I thought one looked very similar to the FEI Technai series, and the other I can't remember the name of, but it has the smoked plastic around the column.


Steve Lee
Chief Technologist
Electron Microscopy Laboratory
Baylor University Medical Center
3500 Gaston Ave
Dallas, TX 75246
( 214.820.3302
Ê 214.820.4110
( stevenle-at-baylorhealth.edu


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Wednesday, January 27, 2010 1:57 PM
To: Lee, Steven

I am pretty sure I saw it too. At least a very TEM-like column.


Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




-----Original Message-----
X-from: jae5-at-lehigh.edu [mailto:jae5-at-lehigh.edu]
Sent: Wednesday, January 27, 2010 1:50 PM
To: Tindall, Randy D.


I know that some people collect sightings of electron microscopes in
movies. Am I the only person who was so bored by Avatar that I spotted
a TEM (at least one, it might even have been two - I will have to wait
for the cable release to be sure) in one of the scenes in the lab?

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


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From: oshel1pe-at-cmich.edu
Date: Wed, 27 Jan 2010 14:39:16 -0600
Subject: [Microscopy] myelinated axons not staining TEM

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


==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
==========================================================

} Date: Wed, 27 Jan 2010 12:15:15 -0800 (PST)
} From: "Susan C. Van Horn" {susan.vanhorn-at-sunysb.edu}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Wednesday, January
} 27, 2010 at 12:15:13 PM.
}
} realname - Susan C. Van Horn
} Email - susan.vanhorn-at-sunysb.edu
} EDUCATION - Graduate College
} LOCATION - Stony Brook
} SUBJECT_OF_QUESTION - myelinated axons
} QUESTION - i am having trouble with myelinated axons essentially
} dropping out - no electron dense showing when view with the TEM and
} even the stained thick sections show a drop out of stain......it
} doesn't seem to matter if it is brain, spinal cord, optic nerve
} samples.....could it be a pH problem??? with fix or buffer?? - i am
} assured that the pH is checked but one never knows - i just have
} never seen this before.....any suggestions???
} thanks
} sue
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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From: kehler-at-twu.ca
Date: Wed, 27 Jan 2010 14:53:51 -0600
Subject: [Microscopy] TEM in Avatar

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


We provided a Philips EM 300 as set dressing to the small-screen version of The Andromeda Strain.
I've never managed to see the whole thing through to find out how (or if) they used/abused it.


Darcy Kehler
Sr. Lab Supervisor, Biology | Faculty of Natural and Applied Sciences
Trinity Western University | p: 604.888.7511 ext. 3249
Langley, BC, Canada,
V2Y 1Y1






-----Original Message-----
X-from: george.theodossiou-at-st-annes.ox.ac.uk [mailto:george.theodossiou-at-st-annes.ox.ac.uk]
Sent: Wednesday, January 27, 2010 12:43 PM
To: Darcy Kehler

There is also an orange column TEM in the TV series Dexter. When he's in his little lab.
I too have been bored by TV and movies.

More TEM's in movies I say....actually how bout an old VG HB501. I know the one here has more character and versatility than some leading actors and actresses out there and it works for free. The microscopist on the other hand will need food, drink and board.

________________________________________
X-from: StevenLe-at-BaylorHealth.edu [StevenLe-at-BaylorHealth.edu]
Sent: 27 January 2010 20:08
To: George Theodossiou

I saw them as well. I thought one looked very similar to the FEI Technai series, and the other I can't remember the name of, but it has the smoked plastic around the column.


Steve Lee
Chief Technologist
Electron Microscopy Laboratory
Baylor University Medical Center
3500 Gaston Ave
Dallas, TX 75246
( 214.820.3302
Ê 214.820.4110
( stevenle-at-baylorhealth.edu


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Wednesday, January 27, 2010 1:57 PM
To: Lee, Steven

I am pretty sure I saw it too. At least a very TEM-like column.


Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




-----Original Message-----
X-from: jae5-at-lehigh.edu [mailto:jae5-at-lehigh.edu]
Sent: Wednesday, January 27, 2010 1:50 PM
To: Tindall, Randy D.


I know that some people collect sightings of electron microscopes in movies. Am I the only person who was so bored by Avatar that I spotted a TEM (at least one, it might even have been two - I will have to wait for the cable release to be sure) in one of the scenes in the lab?

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


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From: lee-at-asc.magnet.fsu.edu
Date: Wed, 27 Jan 2010 15:46:14 -0600
Subject: [Microscopy] Updated: TEM Laboratory Support Scientist Position Open at FSU

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

There was a delay in this position being available for on-line application
but that has now been corrected. We apologize to those of you that went
searching for this position and could not find it at the at the FSU site
listed below.


Job Title: Advanced Analytical TEM Laboratory Support Scientist
Job ID: 31065
Location: National High Magnetic Field Laboratory at Florida State
University, Tallahassee, FL

In conjunction with the recent purchase of a new JEM-ARM200F Atomic
Resolution Analytical TEM we have an opening for a TEM Laboratory Support
Scientist.

Qualifications: Advanced University degree(Master's or higher) and at
least four years of demonstrated experience.

Responsibilities:

*Interacting, assisting and training users for routine operation of the
TEM/STEM microscopes and related sample preparation equipment.
*Performing routine maintenance and essential repair of the microscopes
and related sample preparation equipment (major equipment is under service
contract).
*Managing routine operation, including logs, bookkeeping, billing,
maintenance records, inventory, and consumables purchases.
*Making recommendations and assist in the purchase of new equipment.
*Participating in the microscopy community and related research
activities.
*Preparing monthly and annual reports concerning the TEM Laboratory and
participate in all required review meetings.
*Providing outreach and tours for the general public.

More information and the on-line application (Job ID 31065) is available
at https://jobs.fsu.edu
The NHMFL web site can be found at: http://www.magnet.fsu.edu/
A partial listing of existing analytical facilities at NHMFL can be found
at:
http://www.magnet.fsu.edu/magnettechnology/research/materials/microanalysi
s/index.html

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From: donald.gibbon-at-matcoinc.com
Date: Wed, 27 Jan 2010 15:49:15 -0600
Subject: [Microscopy] Re: Liquid Nitrogen Safety

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

Someone mentioned tactile feedback in hand-cranked ice-cream making. One
of the fundamental correlations in our serious investigations of the
thermodynamics of ice-cream making was between pulse-rate of the cranker
and the viscosity of the ice-cream... leading to our knowing when to
stop cranking and start eating!

Another fundamental property of good ice-cream is called "over-run" (as
I remember, though WHY it's called this I don't recall!). Any way, it's
the incorporation of air bubbles in the product, making it feel
"creamier" on the tongue. When you unfortunately can't eat all the
product immediately and have to try to save some in the freezer
compartment of your refrigerator, it's never as good. The crystals grow
and it may also lose the air bubbles, making it far less luxurious than
it was right after it was made.

I'm not sure how this is different than the fact that on the
pseudo-milk-shake machines in fast-food restaurants there is a control
to determine how much air to put into the shake. I noticed many years
ago that my shakes were getting lighter in weight ... and abandoned that
as a complete offense to my sense of right and wrong! If proper milk
shakes are made from whole, rich ice cream and good whole milk, the air
gets in them from the frothing action of the "turbine" ... and that's
OK!

My patron saints in this whole area are Escoffier, Brillat Savrin, Julia
Child and Harold McGee, whose book, "On Food and Cooking: The Science
and Lore of the Kitchen" is irreplaceable!

Donald L. Gibbon

-----Original Message-----
X-from: paul_hazelton-at-umanitoba.ca [mailto:paul_hazelton-at-umanitoba.ca]
Sent: Wednesday, January 27, 2010 10:38 AM
To: Gibbon, Donald L.

Justin

Escoffier said that a recipe is only a list of ideas. So no, science
has not impacted on my cooking. I still have some great successes in
the kitchen, and some truly magnificent failures. However, science sure

has had an impact on my wine making, and for the better.

paul

--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 William Avenue
Winnipeg, Manitoba, Canada, R3E 0J9
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926



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From: rigler-at-ett.bme.hu
Date: Wed, 27 Jan 2010 17:17:04 -0600
Subject: [Microscopy] [Filtered] MicroscopyListserverviaWWW: SEM - crossover image

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

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: rigler-at-ett.bme.hu
Name: Daniel Rigler

Organization: engineer, failure analysis expert

Title-Subject: [Filtered] SEM - crossover

Message: Hi!

Can somebody please tell me a little bit about how a SEM is capable
of displaying an image of the crossover? I use an FEI Inspect S50
microscope, and wonder what is the detection mechanism.

Thanks,
Daniel

Login Host: 195.56.212.25
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==============================Original Headers==============================
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8, 11 -- From: rigler-at-ett.bme.hu (by way of MicroscopyListserver)
8, 11 -- Subject: [Filtered] MicroscopyListserverviaWWW: SEM - crossover image
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From: analytic-at-rawbw.com
Date: Wed, 27 Jan 2010 19:43:34 -0600
Subject: [Microscopy] viaWWW: SEM at Montpelier High School

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

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: analytic-at-rawbw.com
Name: Margo

Organization: MSA

Title-Subject: [Filtered] High School SEM

Message: I would like to find out if anyone knows anything about the
SEM at Montpelier High School Montpelier, Vermont
ref:
Montpelier is benefiting from a confluence of circumstances, ... the
donation of a scanning electron microscope (SEM) from IBM of Essex
Junction. ... IBM was "retiring" this particular microscope and
donated it to Montpelier High School ...


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From: Aleksandr.Mironov-at-manchester.ac.uk
Date: Thu, 28 Jan 2010 05:46:30 -0600
Subject: [Microscopy] TEM: FEI Polara Facility in Manchester (UK)

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

Dear colleagues,

The University of Manchester Faculty of Life Science EM facility is
pleased to announce the installation of a 300 KeV FEG cryo-transmission
electron microscope funded by the Wellcome Trust. The microscope is
available for use by University of Manchester staff and external
collaborators who have projects that would benefit from collection of
high resolution electron microscope images at cryogenic temperatures.
The microscope is equipped with a 4k x 4k Gatan USC 4000 camera and a
post column energy filter with a 2k x 2k CCD camera. A variety of
software packages are available, both on the microscope for data
generation and off the microscope for data analysis. These include
tomographic packages: FEI’s 3D Explore, SerialEM with IMOD and a variety
of single particle packages, including: SPIDER Appion and MRC. We are
continuously reviewing the installed software, so we hope to be able to
offer the most up to date acquisition and microscope scripting
potential. The FLS electron microscope facility is also able to offer
technical support in order that users are able get the best data from
their samples. If you are interested in using the Polara then please
visit http://manchesterpolara.org.uk, or email polara-at-manchester.ac.uk

Sincerely,
Alex
--
Dr. Aleksandr Mironov MD, PhD
Experimental Officer
D.1527, M.Smith Building
EM Core Facility, Faculty of Life Sciences
University of Manchester
Oxford Road
Manchester
M13 9PT
UK

Tel. +44-(0)161-275-5645
Fax. +44-(0)161-275-5171
E-mail: Aleksandr.Mironov-at-manchester.ac.uk
http://www.ls.manchester.ac.uk/research/facilities/electronmicroscopy/


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From: Frank_Karl-at-lincolnelectric.com
Date: Thu, 28 Jan 2010 06:29:34 -0600
Subject: [Microscopy] Re: Liquid Nitrogen Safety

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

-My patron saints in this whole area are Escoffier, Brillat Savrin, Julia
-Child and Harold McGee, whose book, "On Food and Cooking: The Science
-and Lore of the Kitchen" is irreplaceable!

Let me add that Arthur Grosser's "Cookbook Decoder or Culinary Alchemy
Explained" is an excellent starting place for understanding the chemistry
behind cooking.

On the ice cream front, several years ago I was at the Ohio State Fair
(collecting feather and hair reference samples) and someone was trying to
sell franchises for liquid N2 made ice cream. It didn't seem to me
feasible to have the additional cost of LN2 added to the other cost in what
appears in Akron, to be a saturated market. Maybe that's why I'm working
for someone else.....

stay safe.......
Frank




donald.gibbon-at-mat
coinc.com
To
01/27/2010 04:57 frank_karl-at-lincolnelectric.com
PM cc

Subject
Please respond to [Microscopy] Liquid Nitrogen
donald.gibbon-at-mat Safety
coinc.com












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


Someone mentioned tactile feedback in hand-cranked ice-cream making. One
of the fundamental correlations in our serious investigations of the
thermodynamics of ice-cream making was between pulse-rate of the cranker
and the viscosity of the ice-cream... leading to our knowing when to
stop cranking and start eating!

Another fundamental property of good ice-cream is called "over-run" (as
I remember, though WHY it's called this I don't recall!). Any way, it's
the incorporation of air bubbles in the product, making it feel
"creamier" on the tongue. When you unfortunately can't eat all the
product immediately and have to try to save some in the freezer
compartment of your refrigerator, it's never as good. The crystals grow
and it may also lose the air bubbles, making it far less luxurious than
it was right after it was made.

I'm not sure how this is different than the fact that on the
pseudo-milk-shake machines in fast-food restaurants there is a control
to determine how much air to put into the shake. I noticed many years
ago that my shakes were getting lighter in weight ... and abandoned that
as a complete offense to my sense of right and wrong! If proper milk
shakes are made from whole, rich ice cream and good whole milk, the air
gets in them from the frothing action of the "turbine" ... and that's
OK!

My patron saints in this whole area are Escoffier, Brillat Savrin, Julia
Child and Harold McGee, whose book, "On Food and Cooking: The Science
and Lore of the Kitchen" is irreplaceable!

Donald L. Gibbon

-----Original Message-----
X-from: paul_hazelton-at-umanitoba.ca [mailto:paul_hazelton-at-umanitoba.ca]
Sent: Wednesday, January 27, 2010 10:38 AM
To: Gibbon, Donald L.

Justin

Escoffier said that a recipe is only a list of ideas. So no, science
has not impacted on my cooking. I still have some great successes in
the kitchen, and some truly magnificent failures. However, science sure

has had an impact on my wine making, and for the better.

paul

--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 William Avenue
Winnipeg, Manitoba, Canada, R3E 0J9
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926



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For more information about Valmont Industries, Inc., please visit our web
site at
www.valmont.com


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20, 27 -- In-Reply-To: {201001271537.o0RFbrwL020199-at-ns.microscopy.com}
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20, 27 -- Thread-Topic: [Microscopy] Re: Liquid Nitrogen Safety
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20, 27 -- References: {201001271537.o0RFbrwL020199-at-ns.microscopy.com}
20, 27 -- From: "Gibbon, Donald L." {donald.gibbon-at-matcoinc.com}
20, 27 -- To: {paul_hazelton-at-umanitoba.ca} , {microscopy-at-microscopy.com}
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==============================Original Headers==============================
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From: bengu-at-fen.bilkent.edu.tr
Date: Thu, 28 Jan 2010 07:02:43 -0600
Subject: [Microscopy] [SEM] Experiences on Peltier Stages

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


Dear List,

I would like to hear your experiences on various Peltier Stages for wet
applications (pros/con, dos/donts). Also, I would appreciate any
documentation/procedure for VP mode (air/H2O) for an CZ EVO40 XVP. You can
respond directly to me to save bandwidth for the list

Best

Erman Bengu

=================================
Erman Bengu

Assistant Professor of Chemistry
Department of Chemistry
Bilkent University

Mailing Address:
Bilkent University,
Department of Chemistry,
06800, Bilkent, Ankara
Turkey

Office: SB #311
E-mail: bengu_AT_fen.bilkent.edu.tr
Phone (Office): +90 (312) 290-2153
(Lab1): +90 (312) 290-2663
(Lab2): +90 (312) 290-3332
Fax: +90 (312) 266-4068
Web: http://www.fen.bilkent.edu.tr/~bengu
==================================





==============================Original Headers==============================
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13, 22 -- Date: Thu, 28 Jan 2010 15:02:59 +0200 (EET)
13, 22 -- Subject: [SEM] Experiences on Peltier Stages
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From: paul_hazelton-at-umanitoba.ca
Date: Thu, 28 Jan 2010 08:45:48 -0600
Subject: [Microscopy] Liquid Nitrogen Safety

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

Gee, now that we're in to it, how can we forget Paul Bocuse or Jacques
Pepin.

Although I'm not sure either would add liquid nitrogen to their
kitchens. But then, before this week, neither would I have. Just
can't wait to get the HIV group's students next summer.....(everyone can
now insert an in evil laugh at this spot).

paul

--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 William Avenue
Winnipeg, Manitoba, Canada, R3E 0J9
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926



==============================Original Headers==============================
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From: Ronnie.Reyes-at-Amedd.Army.Mil
Date: Thu, 28 Jan 2010 09:10:05 -0600
Subject: [Microscopy] viaWWW: JOB ANNOUNCEMENT (POSITION: MICROBIOLOGIST) USAMRIID

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


This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: Ronnie.Reyes-at-Amedd.Army.Mil
Name: Ronnie.Reyes

Organization: United States Army Medical Research Institute of
Infectious Diseases (USAMRIID), Fort Detrick, MD

Title-Subject: JOB ANNOUNCEMENT (POSITION: MICROBIOLOGIST)

Message:

DUTIES: An experienced Electron Microscopist is required to plan,
conduct, and provide the technical management of the electron
microscopy portions of USAMRIID studies and in the interpretation of
ultrastructural changes in study materials at the cellular and tissue
levels. Most studies involve research on the pathology of bacterial
and viral pathogens and toxins of interest to the military. A
successful candidate must demonstrate the ability to provide
expertise in the planning and execution of electron microscopy
methodologies and the ability to interpret normal and abnormal
ultrastructural tissue and cellular changes related to a variety of
conditions. The candidate must have excellent written and oral
communication skills, strong project management skills, and the
ability to work cooperatively in a collaborative, cross-functional
team environment.

URL = http://acpol.army.mil/employment/

==============================Original Headers==============================
8, 11 -- From zaluzec-at-microscopy.com Thu Jan 28 09:10:05 2010
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8, 11 -- From: Ronnie.Reyes-at-Amedd.Army.Mil (by way of MicroscopyListserver)
8, 11 -- Subject: viaWWW: JOB ANNOUNCEMENT (POSITION: MICROBIOLOGIST) USAMRIID
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From: Elliott-at-arizona.edu
Date: Fri, 29 Jan 2010 16:42:17 -0600
Subject: [Microscopy] Re: electronic calendar

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

I like OnCore Scheduler. It was developed in-house, but is free for
all. We use it for many of our rooms and 'scopes.
http://demo.arl.arizona.edu
It's easy to download, instal and customize.
David


On Jan 26, 2010, at 1:51 PM, sawyert-at-science.oregonstate.edu wrote:

}
}
}
} ----------------------------------------------------------------------------
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}
} We are interested in setting up an electron scheduling calendar for
} our
} EM lab. Currently we have four microscopes and some sample preparation
} tools users need to sign-up to use. FOM (fomnetworks.com) has been
} suggested but I am wondering about other programs that might be
} cheaper
} or free and still offer similar benefits: scheduling, tracking beam
} time
} etc.
}
} Please offer any suggestions off line
}
} Thanks
} Teresa
}
}
}
}
}
}
}
} --
} Teresa Sawyer
} Electron Microscope Facility Manager
} Oregon State University
} 541-737-5245
} sawyert-at-science.oregonstate.edu
} Cordely Hall 1078
} http://www.science.oregonstate.edu/bpp/EMfacility/index.htm
}
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} ==============================Original
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From: Elliott-at-arizona.edu
Date: Fri, 29 Jan 2010 16:50:57 -0600
Subject: [Microscopy] Re: Science and cooking

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

I find the two very similar.
Beginners should follow the recipe/protocol.
Intermediate practitioners can modify existing recipe/protocol.
Masters can start from scratch and come up with something truly
extraordinary (good or bad :-).

I just try not to mix the ingredients between the two!!

David


On Jan 27, 2010, at 8:59 AM, Chris.Guerin-at-dmbr.vib-UGent.be wrote:

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} ----------------------------------------------------------------------------
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} Hi All:
}
} At the risk of wasting time with off topic posts I have to chime in.
}
} Having been both a professional chef and a scientist I think I can
} say with some authority that if you are good at one chances are you
} will be good at the other, at least in as much as wet lab work goes.
} The combination of being able to follow a protocol (recipe) while at
} the same time having enough creativity and knowledge of the
} principles to experiment a bit and perhaps improve upon it are very
} advantageous in both the lab and the kitchen. Add to that some
} manual dexterity, patience and a sense of genuine joy in the
} possibility of achieving something great and you will get either a
} very good cook (and) or a very good scientist. Whenever I interview
} someone for a technical post I ask if they like to cook, those that
} do generally work out well in the lab.
}
} I will also share that my first kitchen job was as a prep cook. For
} those who don't know a prep cook is the sorry creature who gets to
} do all the messy, difficult and downright disgusting jobs the chef
} no longer wants to do; so you see, it was also excellent training
} for being a grad student.
}
} Best wishes, Chris
}
} Christopher Guérin, Ph.D.
} Leader, Microscopy Core
} Dept. for Molecular Biomedical Research
} Flanders Institute of Biotechnology (VIB)
} University of Ghent, Belgium
}
} ==============================Original
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From: victoria.m.bryg-at-nasa.gov
Date: Fri, 29 Jan 2010 20:10:05 -0600
Subject: [Microscopy] viaWWW: SEM of glass filters

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Name: Vicky Bryg

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Title-Subject: [Filtered] SEM of glass filters

Message: OK. I have some glass filters that I want to do SEM on. I
know they will charge like mad unless I use extremely low KV.
However, my 'free' SEM is limited to one that is not able to be used
at that voltage. Over the years, I have looked at these types of
filters but I was wonder if there are there any new techniques out
there? I don't need high magnification.
Thanks,
Vicky

Victoria M. Bryg
Research Associate
NCSER at NASA Glenn
(216)433-9628
21000 Brookpark Rd.
Cleveland, OH 44135

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From: wesaia-at-iastate.edu
Date: Sun, 31 Jan 2010 18:54:54 -0600
Subject: [Microscopy] viaWWW: SEM of glass filters

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Given a choice, I would use a variable pressure SEM starting at 60 Pa of residual atmosphere (my choice is helium). That should allow imaging at moderate voltages of 15-20 kV using backscattered imaging (i.e., moderate beam currents). We do it routinely and can achieve magnifications of a few thousand times.

Of course, that presume you have such an SEM available. Aside from that, you are probably left at using lower voltages to do what you can. Perhaps tilting the specimen can increase the yield of secondary electrons from the surface so that such low voltages are not required.
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Email: victoria.m.bryg-at-nasa.gov
Name: Vicky Bryg

Organization: NCSER at NASA Glenn

Title-Subject: [Filtered] SEM of glass filters

Message: OK. I have some glass filters that I want to do SEM on. I
know they will charge like mad unless I use extremely low KV.
However, my 'free' SEM is limited to one that is not able to be used
at that voltage. Over the years, I have looked at these types of
filters but I was wonder if there are there any new techniques out
there? I don't need high magnification.
Thanks,
Vicky

Victoria M. Bryg
Research Associate
NCSER at NASA Glenn
(216)433-9628
21000 Brookpark Rd.
Cleveland, OH 44135

Login Host: 128.156.10.80
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From: gary-at-gaugler.com
Date: Sun, 31 Jan 2010 20:26:46 -0600
Subject: [Microscopy] Re: viaWWW: SEM of glass filters

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Can you coat them with Au/Pd or some other sputter
coated metal? It is thin and can be wiped off after
use/analysis. I would not recommend pure Au. Pd is
probably better...IMO.

It would seem that the main issue is to remove the
coating without affecting the optical quality of the
glass. Perhaps you could try a sacrificial specimen and see if
this works. I have done this on cover slips without
any problems. But of course, these are different specimens.

gary g.


At 06:12 PM 1/29/2010, you wrote:



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From: vray-at-partbeamsystech.com
Date: Sun, 31 Jan 2010 23:19:48 -0600
Subject: [Microscopy] Re: viaWWW: SEM of glass filters

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Hi Victoria,

By the time I connect to grid you probably will receive multiple
responses with similar suggestions, but anyway: from my somewhat limited
experience of imaging photolithography masks (Quartz substrate charges
like mad and retains charge under the surface) E-SEM in wet mode has
inherent charge neutralization mechanism which works beautifully for
dielectric materials. I am sure you could find an E-SEM in one of the
universities nearby...

Good Luck :)
Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com

victoria.m.bryg-at-nasa.gov wrote:
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} Title-Subject: [Filtered] SEM of glass filters
}
} Message: OK. I have some glass filters that I want to do SEM on. I
} know they will charge like mad unless I use extremely low KV.
} However, my 'free' SEM is limited to one that is not able to be used
} at that voltage. Over the years, I have looked at these types of
} filters but I was wonder if there are there any new techniques out
} there? I don't need high magnification.
} Thanks,
} Vicky
}
} Victoria M. Bryg
} Research Associate
} NCSER at NASA Glenn
} (216)433-9628
} 21000 Brookpark Rd.
} Cleveland, OH 44135
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From: Frank_Karl-at-lincolnelectric.com
Date: Mon, 1 Feb 2010 06:13:52 -0600
Subject: [Microscopy] Re: viaWWW: SEM of glass filters

Contents Retrieved from Microscopy Listserver Archives
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Hi Vicky,
All good suggestions.
In the past I've used sliver paint to ground the sample to the stud but you
need enough sample you can discard the bad preps. When the viscosity of
the drying paint is just about right, it will wet about half the thickness
of your sample. That the goal. I gold coat the sample twice, once with
the sample facing the source and once with the sample at an angle to the
source. I try to get conductive material all the way around my fibers but
I never succeed completely. I don't like to use a lot of gold on any
sample, so for me, a couple seconds of normal operation is fine. Your
samples may need more. Lowering your Kv always helps, even it's just to 15
from 25kv.

It seems that just pumping the SEM chamber down, causes the glass fibers to
shift, exposing non-conductive surfaces no matter what you do.

Frankly, I also learned to live with a certain amount of charging. It's a
trade off between my goals, the equipment available and the amount of work
I need to accomplish these goals. Many times I just want to reduce the
charging to see my particles and keep them from flying off the sample.
Other times I need a image for a report so the amount of work I put into it
changes.


Low vacuum SEM, if you can live with backscatter electron images works well
on charging plastic, but I don't know how it will work on haystacks of
glass fibers.

stay safe........
Frank



victoria.m.bryg-at-n
asa.gov
To
01/29/2010 09:20 frank_karl-at-lincolnelectric.com
PM cc

Subject
Please respond to [Microscopy] viaWWW: SEM of glass
victoria.m.bryg-at-n filters
asa.gov












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Name: Vicky Bryg

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Title-Subject: [Filtered] SEM of glass filters

Message: OK. I have some glass filters that I want to do SEM on. I
know they will charge like mad unless I use extremely low KV.
However, my 'free' SEM is limited to one that is not able to be used
at that voltage. Over the years, I have looked at these types of
filters but I was wonder if there are there any new techniques out
there? I don't need high magnification.
Thanks,
Vicky

Victoria M. Bryg
Research Associate
NCSER at NASA Glenn
(216)433-9628
21000 Brookpark Rd.
Cleveland, OH 44135

Login Host: 128.156.10.80
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From: eschumacher-at-mccrone.com
Date: Mon, 1 Feb 2010 07:37:36 -0600
Subject: [Microscopy] Short Course Announcement: Transmission Electron Microscopy

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

Greetings Fellow Microscopists,

The College of Microscopy, located in Westmont, IL, is offering a short course in Transmission Electron Microscopy March 23-25, 2010. In addition to lectures, this course emphasizes hands-on training using state of the art equipment. For further details and registration information, please follow the link below:

www.collegeofmicroscopy.com

Regards,

Elaine

*********************************************************************
Elaine F.Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com



*********************************************************************
This message and any attachments are solely for the
intended recipient. If you are not the intended recipient,
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==============================Original Headers==============================
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10, 23 -- Date: Mon, 1 Feb 2010 07:37:31 -0600
10, 23 -- Subject: Short Course Announcement: Transmission Electron Microscopy
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From: allison.van-de-meene-at-bbsrc.ac.uk
Date: Tue, 2 Feb 2010 03:09:16 -0600
Subject: [Microscopy] Cryo EM course 21-26 March 2010, UK

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

Cool Snaps - A course in Cryo Techniques for Electron Microscopy sponsored by the Royal Microscopical Society (RMS) held at Rothamsted Research, Harpenden, Hertfordshire, UK (30 mins north of London).

The intensive course includes teaching sample preparation and cryo-microscopy techniques for both scanning and transmission electron microscopy. Participants from both academia and industry who wish to learn new skills or update their knowledge are encouraged to attend.

Speakers:
Plenary speaker - Kent MacDonald (University of California Berkeley)
Cryofixation - Roger Wepf (ETH Zurich, EMEZ)
Cryo sectioning - Helmut Gnaegi (Diatome)
CryoSEM - Kim Findlay (John Innes Centre)
CryoTEM - Dave Bhella (University of Glasgow)
Freeze substitution and sample processing - Paul Verkade (University of Bristol)
Optimising camera and software variables - Neil Wilkinson (Gatan)
Common artifacts - Marilyn Carey (Gatan)

Early registration ends 12th February 2010.

For further information please look at http://www.rms.org.uk/events/Forthcoming_Events/Cryo_EM_Course.

Hope to see you there!


Dr. Allison van de Meene
Head of Bioimaging
Bioimaging Facility
Plant Pathogen and Microbiology Department
Rothamsted Research
Harpenden
Hertfordshire AL5 2JQ
UK

Tel: +44 1582 763133 ext 2285
Fax: +44 1582 760981
Email: allison.van-de-meene-at-bbsrc.ac.uk
http://www.rothamsted.bbsrc.ac.uk/

===================================
Rothamsted Research is a company limited by guarantee, registered in England at the above address under the registration number 2393175 and a not for profit charity number 802038.
 




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From: ian.drucker-at-gmail.com
Date: Tue, 2 Feb 2010 09:58:49 -0600
Subject: [Microscopy] Metal Grain Size and Count Software?

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

I'm looking for suggestions on software that can automatically count
total grains and also do size analysis. The images would be from a FIB
and be of
various metal samples.

Any ideas out there?


Thanks

==============================Original Headers==============================
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From: bloos-at-sun.ac.za
Date: Tue, 2 Feb 2010 19:21:17 -0600
Subject: [Microscopy] viaWWW: tissue preservation for TEM analysis

Contents Retrieved from Microscopy Listserver Archives
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This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at
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Email: bloos-at-sun.ac.za
Name: Ben Loos

Organization: Stellenbosch University

Title-Subject: [Filtered] tissue preservation for TEM analysis

Message: Dear collegues,

Could you please advice me on how to preserve tissue for TEM
analysis? I have to store the tissue for at least 4 weeks, before
starting with the TEM work. In that time period, should the tissue be
stored in paraformaldehyde or any other fixative, or should it be
stored in the embedding medium itself?

Thanks alot for your advice
best wishes
Ben

Dr Ben Loos
Cell Imaging Unit -Fluorescence Live Cell Imaging & Flow Cytometry-
Central Analytical Facility-CAF
Stellenbosch University - South Africa
Department of Physiological Sciences
Mike de Vries Building
Office 2022
7600 Stellenbosch





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13, 11 -- Subject: viaWWW: tissue preservation for TEM analysis
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From: a.chuvilin-at-microscopist.ru
Date: Wed, 3 Feb 2010 03:16:19 -0600
Subject: [Microscopy] TEM postdoc position

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

As part of a new project, CIC nanoGUNE Consolider Research Center in
Donostia-San
Sebastian (Basque Country, Spain) is currently looking for a talented
and motivated


Postdoctoral fellow
(Physicist/Materials Scientist/Engineer)

in the field of transmission electron microscopy and related techniques.

CIC nanoGUNE Consolider is a recently established Research &
Development Center
with the mission of conducting basic and applied world-class research
in nanoscience and
nanotechnology. The newly initiated electron microscopy facility will
start operation this
year including a fully equipped FEI Titan 60-300 TEM (X-FEG, MC,
imaging Cs-
corrector, EDX, bi-prism, Quantum GIF), a FEI Helios NanoLab FIB
system and a FEI
Quanta 250 FEG ESEM.
The postdoctoral position will be related to exploring the potential
of exciting new
areas in the field of high-end electron microscopy, FIB-based
nanostructure fabrication,
as well as TEM sample preparation. As there will be an intensive work
on FEI systems
and solutions, the candidate will have a unique opportunity to acquire
deep knowledge of
instrumentation and techniques, as well as to learn diverse aspects of
nanotechnology in
interaction with nanoGUNE´s international research team.
Responsibilities of the position will include planning and performing
of experiments,
processing and evaluating of results, preparing publications and
interaction with FEI
specialists. The outcome of the work is expected to be in form of
application reports as
well as scientific publications in internationally leading journals.
The position holder will
be expected to provide expert advice on the application of FEI
instruments, demonstrate
their operation and develop further techniques for sample preparation
and analysis.
The position requires a PhD in Chemistry, Material Science,
Engineering or Physics
strongly related to TEM investigations. The successful candidate will
have very
substantial experimental TEM expertise, a solid theoretical background
in the field of
TEM, experience in FIB operation, as well as good communication and
writing skills.
Proficiency in the following methods is most desirable: CTEM, HRTEM,
STEM, SAED
and CBED, EDX and EELS analysis, as well as image calculations.
Experience in
operating of Cs-corrected instruments is a big advantage.
The position is available from May 1st 2010 for duration of two years.
Interested
applicants should send a letter of application, curriculum vitae, and
names and contact
information of three references to: nano-at-nanogune.eu with the subject
line:
“TEM postdoc application”.

Deadline for application is March 15th, 2010.






--------------------------------
Andrey Chuvilin
Ikerbasque Research Professor
TEM Staff Scientist

CIC nanoGUNE Consolider
Tolosa Hiribidea, 76
E-20018 Donostia - San Sebastian
Spain
+34 943 574 023
www.nanogune.eu
--------------------------------


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From: Ann-Fook.Yang-at-AGR.GC.CA
Date: Wed, 3 Feb 2010 12:24:24 -0600
Subject: [Microscopy] circuit diagram for Hexland CryoTrans 1000

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

Hi Lister,

I need a circuit diagram for a HEXLAND cryostage (CRYOTRANS CT 1000); I
would appreciate it if anyone who has the instrument manual sends a copy
to me. Thank you.
Ann-Fook Yang,



==============================Original Headers==============================
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From: matthew.weyland-at-mcem.monash.edu.au
Date: Wed, 3 Feb 2010 16:08:19 -0600
Subject: [Microscopy] Post doctoral research assistantship @ Monash University, Melbourne

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

=======================

Ref No: A1010515

RESEARCH ASSISTANT IN ELECTRON TOMOGRAPHY OF FIELDS AND POTENTIALS
SCHOOL OF PHYSICS, MONASH UNIVERSITY, AUSTRALIA

Duration: Three-year appointment

Location: School of Physics, Monash University, Australia
& Monash Centre for Electron Microscopy (MCEM)

Applications Close: Friday, 19 February 2010

For further details, see:

http://monash.turborecruit.com.au/job/job_details.cfm?id=435492&from=

Dr M.Weyland, Senior Research Fellow & Electron Microscopist
----------------------------------------------------------------------------
|Monash Centre for Electron Microscopy |Department of Materials Engineering|
|Bldg 81 |Room 115, Bldg 69 |
|Monash University |Monash University |
|Victoria 3800 |Victoria 3800 |
|Australia. |Australia |
|--------------------------------------------------------------------------|
|www.mcem.monash.edu.au |www.materials.monash.eu.au |
|--------------------------------------|-----------------------------------|
|Ph: (+61) 3 990 59026 |Fax: (+61) 3 990 53600 |
----------------------------------------------------------------------------

==============================Original Headers==============================
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From: Frank_Karl-at-lincolnelectric.com
Date: Thu, 4 Feb 2010 07:58:42 -0600
Subject: [Microscopy] working distance on Older Leitz Objectives

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


The barbarians I work for trashed out a old Leitz Ortholux (it makes me
sick just write this) metallograhic reflected light scope. All I could
save were the objectives. (No, I couldn't save it, buy it, donate it or
hide it and I'll step outside with anyone who said otherwise.) We may be
able use the objectives on a Vickers hardness machine.

Does any one know the working distance for these objectives? They are RMS
treads and here's the notation on the objectives:

5X/ 0.09
10x/ 0.18
20x /0.35
Fl 50x/0.85
Fl 100x/0.95

thanks in advance........

Frank

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From: Frank_Karl-at-lincolnelectric.com
Date: Thu, 4 Feb 2010 08:35:54 -0600
Subject: [Microscopy] Re: working distance on Older Leitz Objectives

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

Hi Mark,
The reflected light 5 position nosepiece has a small removable lens in the
objective/ocular path marked with an infinity symbol and 223. I suspect
its an infinite corrected optical system.

Thank !!!




"Mark William"
{Markw-at-wyeth.com}
To
02/04/2010 09:22 {Frank_Karl-at-lincolnelectric.com}
AM cc

Subject
Re: [Microscopy] working distance
on Older Leitz Objectives










Frank,

I think these may be older 170mm tube lebgth objectives have a working
distance shorter than what you will see with DIN objectives. The parfocal
distance will also be shoter than DIN. In order to mix and match, you will
need to get adapters to extend the objective. Leitz also made some DIN
objectives and infinity ones as well. Do they list a tube length?

} } } {Frank_Karl-at-lincolnelectric.com} 2/4/2010 9:09 AM } } }



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The barbarians I work for trashed out a old Leitz Ortholux (it makes me
sick just write this) metallograhic reflected light scope. All I could
save were the objectives. (No, I couldn't save it, buy it, donate it or
hide it and I'll step outside with anyone who said otherwise.) We may be
able use the objectives on a Vickers hardness machine.

Does any one know the working distance for these objectives? They are RMS
treads and here's the notation on the objectives:

5X/ 0.09
10x/ 0.18
20x /0.35
Fl 50x/0.85
Fl 100x/0.95

thanks in advance........

Frank

--
*************************************************************
Note:
The information contained in this message may be
privileged and confidential and protected from disclosure. If
the reader of this message is not the intended recipient, or
an employee or agent responsible for delivering this message
to the intended recipient, you are hereby notified that any
dissemination, distribution or copying of this communication
is strictly prohibited. If you have received this
communication in error, please notify us immediately by
replying to the message and deleting it from your computer.
Thank you,
The Lincoln Electric Company
**************************************************************


==============================Original
Headers==============================
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-0600
8, 21 -- Subject: working distance on Older Leitz Objectives
8, 21 -- To: Microscopy-at-microscopy.com
8, 21 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005
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27, 21 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005
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From: msanders-at-umn.edu
Date: Thu, 4 Feb 2010 13:59:18 -0600
Subject: [Microscopy] viaWWW: Microwave Processing Workshop

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This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at
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Email: msanders-at-umn.edu
Name: Mark Sanders

Organization: University of Minnesota, Imaging Center

Title-Subject: [Filtered] Microwave Processing Workshop

Message: I would like to make you aware of an upcoming workshop on
Microwave-assisted processing for both light and electron microscopy.

The 3-day workshop will focus on optimizing techniques using
microwave specimen processing for microscopy.

March 10-12, 2010
University of Minnesota
College of Biological Sciences, Imaging Center
23-35 Snyder Hall
St. Paul, MN 55108

Hands-on and Lecture Topics covered will include:
In vivo Labeling.
Live Cell Imaging using Widefield, Confocal and Spinning Disk
Microscopy. PerkinElmer will have their latest spinning disk system
on hand.
Principles and Application of Microwave Fixation for Biological Materials.
Principles and Application of Immunolocalization techniquies for
Confocal and Fluorescence Microscopy.
In situ Hybridization.

In addition to our fully equipped facility, the latest
instrumentation from PerkinElmer (spinning disk), Ted Pella (BioWave
Pro microwave processor) and Leica (EM AMW processor) will be
available for participants.

For additional information and to register, please point your browser to:
www.cbs.umn.edu/ic/events/mwws10.shtml

Feel free to contact me with questions.

Mark Sanders
Program Director, CBS Imaging Center
msanders-at-umn.edu

Login Host: 160.94.173.227
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From: oshel1pe-at-cmich.edu
Date: Thu, 4 Feb 2010 15:50:31 -0600
Subject: [Microscopy] Fwd: Ask-A-Microscopist

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

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
==========================================================

} Date: Thu, 4 Feb 2010 13:30:43 -0800 (PST)
} From: Daniel Thayer {kanawai-at-hotmail.com}
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Thursday, February
} 04, 2010 at 01:30:36 PM.
}
} realname - Daniel Thayer
} Email - kanawai-at-hotmail.com
} ORGANIZATION - Colorado State University
} EDUCATION - Graduate College
} LOCATION - Fort Collins, Colorado, United States
} SUBJECT_OF_QUESTION - imaging sucrose particles by SEM
} QUESTION - I am collecting sucrose particles on an aluminum plate
} and imaging them with a SEM. I'm fairly new to the process and am
} having trouble finding any particles. I calculated that there should
} be about 23 particles per field at the concentrations generated for
} the test. I am able to bring the image into sharp focus, so I should
} be able to see any particles in the field of view. I'd like to rule
} out the possiblility that the reason for this problem is that the
} particles are evaporating under the electron beam when I try to
} image them. The particles are 15 nm and I am imaging them at 65,000X
} at 15.0 kV. The particles also contain trace amounts of ammonium
} acetate from the buffer solution used while creating them. The
} sample is sputter-coated with 5 nm of gold prior to imaging. Should
} these conditions cause the particles to evaporate before I can focus
} on them?
}
} Thanks,
}
} Daniel
}

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From: smithj-at-winthrop.edu
Date: Thu, 4 Feb 2010 16:14:24 -0600
Subject: [Microscopy] Keeping the moribund alive--Copy of an MT2B service manual?

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

Can provide a copy of an MT5000 manual in return.
Julian

--
Julian P.S. Smith III
Director, Winthrop Microscopy Facility
Dept. of Biology
Winthrop University
520 Cherry Rd.
Rock Hill, SC 29733

803-323-2111 x6427 (vox)
803-323-3448 (fax)
803-524-2347 (cell)


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From: connellyps-at-nhlbi.nih.gov
Date: Thu, 4 Feb 2010 16:31:54 -0600
Subject: [Microscopy] Re: Keeping the moribund alive--Copy of an MT2B

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


Julian,

I have the MT2 manual and there are 3 pages on maintenance. I'll scan them if you want but contact me off line with questions. I've been taking care of the MT2 (and still using one) for several decades. Both models are very similar.

I know of a company in PA who services many thpes of ultra-microtomes if you are in need.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E - Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-480-6560
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov} {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not represent the NIH. This message is not confidential and can be freely shared and reproduced.


________________________________
X-from: {smithj-at-winthrop.edu}

Can provide a copy of an MT5000 manual in return.
Julian

--
Julian P.S. Smith III
Director, Winthrop Microscopy Facility
Dept. of Biology
Winthrop University
520 Cherry Rd.
Rock Hill, SC 29733

803-323-2111 x6427 (vox)
803-323-3448 (fax)
803-524-2347 (cell)




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From: doc.vrdoljak-at-gmail.com
Date: Thu, 4 Feb 2010 17:09:46 -0600
Subject: [Microscopy] cryo TEM for service fee

Contents Retrieved from Microscopy Listserver Archives
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Hello,
I have an emulsion that I'd like to do cryo-TEM on. Does anyone know
of a fee for service lab where I can submit samples to get this kind
of work done?
Thanks.

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From: kenconverse-at-qualityimages.biz
Date: Thu, 4 Feb 2010 17:23:32 -0600
Subject: [Microscopy] Fwd: Ask-A-Microscopist

Contents Retrieved from Microscopy Listserver Archives
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Daniel,
How do you know you're in sharp focus? Unless this is an FESEM, 65kX is
fairly high mag for 15kV. Are you sure you're resolving 15nm? In fact you
probably need to be resolving 10nm or better to have a good idea if you're
seeing your particles.

What is your condenser lens/probe size/beam current setting? If your image
isn't crisp (due to spot size, not focus), you might possibly have enough
beam current to evaporate your sample.

Is there a chance that you haven't actually made particles?

Without a gold coating, they would be very hard to see. I'm assuming that
you're gold coating is in the range that you describe. You might try 10nm
and see if anything changes.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
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kenconverse-at-qualityimages.biz
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-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Thursday, February 04, 2010 4:52 PM
To: kenconverse-at-qualityimages.biz

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
==========================================================

} Date: Thu, 4 Feb 2010 13:30:43 -0800 (PST)
} From: Daniel Thayer {kanawai-at-hotmail.com}
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Thursday, February
} 04, 2010 at 01:30:36 PM.
}
} realname - Daniel Thayer
} Email - kanawai-at-hotmail.com
} ORGANIZATION - Colorado State University
} EDUCATION - Graduate College
} LOCATION - Fort Collins, Colorado, United States
} SUBJECT_OF_QUESTION - imaging sucrose particles by SEM
} QUESTION - I am collecting sucrose particles on an aluminum plate
} and imaging them with a SEM. I'm fairly new to the process and am
} having trouble finding any particles. I calculated that there should
} be about 23 particles per field at the concentrations generated for
} the test. I am able to bring the image into sharp focus, so I should
} be able to see any particles in the field of view. I'd like to rule
} out the possiblility that the reason for this problem is that the
} particles are evaporating under the electron beam when I try to
} image them. The particles are 15 nm and I am imaging them at 65,000X
} at 15.0 kV. The particles also contain trace amounts of ammonium
} acetate from the buffer solution used while creating them. The
} sample is sputter-coated with 5 nm of gold prior to imaging. Should
} these conditions cause the particles to evaporate before I can focus
} on them?
}
} Thanks,
}
} Daniel
}

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From: jquinn-at-www.matscieng.sunysb.edu
Date: Thu, 4 Feb 2010 18:17:21 -0600
Subject: [Microscopy] re: sucrose

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

Daniel and those "Out of Office"

15nm x 65,000 = 975,000nm = 975 microns = 0.975 mm

Basically, your 15nm particle is a speck on the screen.

Next, you are uniformaly coating the specks and substrate
with gold, at 1/3 the speck size.

It is not surprising that you do not see anything.

Especially, since you are using 15kV on a light-element (sucrose)
particle. The scattering volume dwarfs the speck size.

just my two cents,

JQuinn

}
} Date: Thu, 4 Feb 2010 13:30:43 -0800 (PST)
} From: Daniel Thayer {kanawai-at-hotmail.com}
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Thursday, February
} 04, 2010 at 01:30:36 PM.
}
} realname - Daniel Thayer
} Email - kanawai-at-hotmail.com
} ORGANIZATION - Colorado State University
} EDUCATION - Graduate College
} LOCATION - Fort Collins, Colorado, United States
} SUBJECT_OF_QUESTION - imaging sucrose particles by SEM
} QUESTION - I am collecting sucrose particles on an aluminum plate
} and imaging them with a SEM. I'm fairly new to the process and am
} having trouble finding any particles. I calculated that there should
} be about 23 particles per field at the concentrations generated for
} the test. I am able to bring the image into sharp focus, so I should
} be able to see any particles in the field of view. I'd like to rule
} out the possiblility that the reason for this problem is that the
} particles are evaporating under the electron beam when I try to
} image them. The particles are 15 nm and I am imaging them at 65,000X
} at 15.0 kV. The particles also contain trace amounts of ammonium
} acetate from the buffer solution used while creating them. The
} sample is sputter-coated with 5 nm of gold prior to imaging. Should
} these conditions cause the particles to evaporate before I can focus
} on them?
}
} Thanks,
}
} Daniel
}

==============================Original Headers==============================
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9, 12 -- Message-Id: {201002050011.o150BUX30342-at-www.matscieng.sunysb.edu}
9, 12 -- To: microscopy-at-microscopy.com
9, 12 -- Subject: re: sucrose
==============================End of - Headers==============================





From: lene.cecilie.hermansen-at-veths.no
Date: Fri, 5 Feb 2010 04:32:49 -0600
Subject: [Microscopy] viaWWW: Fixation solution for crustaceans

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Email: lene.cecilie.hermansen-at-veths.no
Name: Lene Cecilie Hermansen

Organization: Norwegian School of Veterinary Science

Title-Subject: [Filtered] Fixation solution for crustaceans

Message: Hi

I wonder if anybody has successfully epon-embedded Artemia, and which
fixation solution did you use?

(Artemia is a small, crustaceans, organism;
http://en.wikipedia.org/wiki/Artemia).

Regards Lene Hermansen,
Norwegian School of Veterinary Science

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From: dbagrov-at-gmail.com
Date: Fri, 5 Feb 2010 05:17:16 -0600
Subject: [Microscopy] TEM - a question about sampling and magnification

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

Hello!
I study nanoparticles by transmission electron microscopy. After
examining several samples I have faced to two questions:
1. What is the minimum magnification which is required to notice a nanoparticle?
2. What is the minimum magnification which is required to describe the
shape of a nanoparticle?
The answer to the first question seems to depend on the particle size,
the resolution (number of pixels) of CCD camera, and probably the
physical size of CCD (according to the Nyquist theorem). Is there a
formula to calculate the minimum required magnification?
The second question seems more tricky than the first one (at least for
me). To my mind, there should be at least two formulas (one for the
minimum size and the other one for the maximum size of a particle).
Could you please help me with these question? I was trying to find
answers on the Internet, but I couldn't find any concise examples,
neither the formulas.
Thanks!
--
Sincerely yours,
Dmitry Bagrov

==============================Original Headers==============================
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1, 32 -- Message-ID: {af91b5e61002050317r7c3755f8h80ef9b9ff396345f-at-mail.gmail.com}
1, 32 -- Subject: TEM - a question about sampling and magnification
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From: nizets2-at-yahoo.com
Date: Fri, 5 Feb 2010 05:43:44 -0600
Subject: [Microscopy] TEM - a question about sampling and magnification

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

The only formula you need is the multiplication sign.
Let's say that 1mm is a "comfortable" size to notice a particle.
if your particle is 10nm in size, you'll need a 100,000x mag to make it appear 1mm big on your screen.
(1 mm=1000 µm=1000,000 nm)
Knowing that, I think you can guess yourself at what apparent size you are able to describe a shape.

Stephane

NB: "comfortable" is probably not the right word, because your particle will still look like a small spot on the screen.



----- Original Message ----
X-from: "dbagrov-at-gmail.com" {dbagrov-at-gmail.com}
To: nizets2-at-yahoo.com
Sent: Fri, February 5, 2010 12:19:55 PM

Hello!
I study nanoparticles by transmission electron microscopy. After
examining several samples I have faced to two questions:
1. What is the minimum magnification which is required to notice a nanoparticle?
2. What is the minimum magnification which is required to describe the
shape of a nanoparticle?
The answer to the first question seems to depend on the particle size,
the resolution (number of pixels) of CCD camera, and probably the
physical size of CCD (according to the Nyquist theorem). Is there a
formula to calculate the minimum required magnification?
The second question seems more tricky than the first one (at least for
me). To my mind, there should be at least two formulas (one for the
minimum size and the other one for the maximum size of a particle).
Could you please help me with these question? I was trying to find
answers on the Internet, but I couldn't find any concise examples,
neither the formulas.
Thanks!
--
Sincerely yours,
Dmitry Bagrov

==============================Original Headers==============================
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1, 32 -- Message-ID: {af91b5e61002050317r7c3755f8h80ef9b9ff396345f-at-mail.gmail.com}
1, 32 -- Subject: TEM - a question about sampling and magnification
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16, 25 -- Subject: Re: [Microscopy] TEM - a question about sampling and magnification
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From: dbagrov-at-gmail.com
Date: Fri, 5 Feb 2010 06:51:52 -0600
Subject: [Microscopy] Re: TEM - a question about sampling and magnification

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

Dear Stephane,
Thanks for your quick reply!
Actually I was asking from a more "theoretical" point of view: the
required magnification doesn't depend ONLY on the particle size and
the size of the screen. One of the reasons is that the resolution of
the microscope is finite and increasing magnification doesn't always
allow to measure the size and analyze the shape of small particles.
Let's say that X is the number of pixels in the CCD camera (typically
1000 or 2000), L is the physical size of the sample which is projected
on the CCD camera. If we increase the magnification, L decreases
proportionally (the analyzed area becomes smaller).
According to the Nyquist theorem the minimum size of particle that can
be noticed is 2L/X. Let's assume that I need 5x5 pixels to describe
the shape of a particle (I am not sure if it is correct). So if the
magnification is constant (L is constant) I will be able to describe
the shape of a particle which is larger than 5L/X. If a microscope
provides a 300x300 nm image with 500'000 magnification and I have a
1000x1000 pixel camera I will be able to describe the shape of
particles which are larger than 5x300 nm/1000=1,5 nm (if the
resolution of the microscope allows me to do so).
Is there a way to make these or similar estimations in general? How
many pixels is needed to describe the shape of a particle?
--
Sincerely yours,
Dmitry Bagrov

2010/2/5 Stephane Nizet {nizets2-at-yahoo.com} :
} The only formula you need is the multiplication sign.
} Let's say that 1mm is a "comfortable" size to notice a particle.
} if your particle is 10nm in size, you'll need a 100,000x mag to make it appear 1mm big on your screen.
} (1 mm=1000 µm=1000,000 nm)
} Knowing that, I think you can guess yourself at what apparent size you are able to describe a shape.
}
} Stephane
}
} NB: "comfortable" is probably not the right word, because your particle will still look like a small spot on the screen.
}
}
}
} ----- Original Message ----
} From: "dbagrov-at-gmail.com" {dbagrov-at-gmail.com}
} To: nizets2-at-yahoo.com
} Sent: Fri, February 5, 2010 12:19:55 PM
} Subject: [Microscopy] TEM - a question about sampling and magnification
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} Hello!
} I study nanoparticles by transmission electron microscopy. After
} examining several samples I have faced to two questions:
} 1. What is the minimum magnification which is required to notice a nanoparticle?
} 2. What is the minimum magnification which is required to describe the
} shape of a nanoparticle?
} The answer to the first question seems to depend on the particle size,
} the resolution (number of pixels) of CCD camera, and probably the
} physical size of CCD (according to the Nyquist theorem). Is there a
} formula to calculate the minimum required magnification?
} The second question seems more tricky than the first one (at least for
} me). To my mind, there should be at least two formulas (one for the
} minimum size and the other one for the maximum size of a particle).
} Could you please help me with these question? I was trying to find
} answers on the Internet, but I couldn't find any concise examples,
} neither the formulas.
} Thanks!
} --
} Sincerely yours,
} Dmitry Bagrov
}
} ==============================Original Headers==============================
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} 1, 32 -- Message-ID: {af91b5e61002050317r7c3755f8h80ef9b9ff396345f-at-mail.gmail.com}
} 1, 32 -- Subject: TEM - a question about sampling and magnification
} 1, 32 -- From: Dmitry Bagrov {dbagrov-at-gmail.com}
} 1, 32 -- To: Microscopy-at-microscopy.com
} 1, 32 -- Content-Type: text/plain; charset=ISO-8859-1
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3, 37 -- Subject: Re: [Microscopy] TEM - a question about sampling and magnification
3, 37 -- From: Dmitry Bagrov {dbagrov-at-gmail.com}
3, 37 -- To: Stephane Nizet {nizets2-at-yahoo.com}
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From: Philip.Koeck-at-ki.se
Date: Fri, 5 Feb 2010 07:39:37 -0600
Subject: [Microscopy] Re: TEM - a question about sampling and magnification

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

Hi,

I would suggest reformulating and simplifying the question.
The size of the particles is not really so important since they are
quite small and you'll easily fit at least one of them in one image.

Decide first at what level of detail you want to image the particles.
That is the spatial resolution you need. Let's say you want 1 nm spatial
resolution. That means that the pixel size mustn't be bigger than 0.5 nm
(according to the sampling theorem). To be on the safe side and minimize
interpolation errors (if you want to perform averaging) you should choose
a pixel size of about 0.3 nm.

If your detector or scanner has a pixel size of for example 25 micrometers
(25000 nm) the magnification between specimen and detector should be
about 25000/0.3=80000.

If you want 0.2 nm resolution you'll need 400000 magnification.

So the formula for the correct magnification is:

detectorpixelsize divided by (desiredresolution times 0.3)

(with everything given in the same units)

Philip

-----Original Message-----
X-from: dbagrov-at-gmail.com [mailto:dbagrov-at-gmail.com]
Sent: 05 February 2010 13:54
To: Philip.Koeck-at-biosci.ki.se

Dear Stephane,
Thanks for your quick reply!
Actually I was asking from a more "theoretical" point of view: the
required magnification doesn't depend ONLY on the particle size and
the size of the screen. One of the reasons is that the resolution of
the microscope is finite and increasing magnification doesn't always
allow to measure the size and analyze the shape of small particles.
Let's say that X is the number of pixels in the CCD camera (typically
1000 or 2000), L is the physical size of the sample which is projected
on the CCD camera. If we increase the magnification, L decreases
proportionally (the analyzed area becomes smaller).
According to the Nyquist theorem the minimum size of particle that can
be noticed is 2L/X. Let's assume that I need 5x5 pixels to describe
the shape of a particle (I am not sure if it is correct). So if the
magnification is constant (L is constant) I will be able to describe
the shape of a particle which is larger than 5L/X. If a microscope
provides a 300x300 nm image with 500'000 magnification and I have a
1000x1000 pixel camera I will be able to describe the shape of
particles which are larger than 5x300 nm/1000=1,5 nm (if the
resolution of the microscope allows me to do so).
Is there a way to make these or similar estimations in general? How
many pixels is needed to describe the shape of a particle?
--
Sincerely yours,
Dmitry Bagrov

2010/2/5 Stephane Nizet {nizets2-at-yahoo.com} :
} The only formula you need is the multiplication sign.
} Let's say that 1mm is a "comfortable" size to notice a particle.
} if your particle is 10nm in size, you'll need a 100,000x mag to make it
appear 1mm big on your screen.
} (1 mm=1000 µm=1000,000 nm)
} Knowing that, I think you can guess yourself at what apparent size you are
able to describe a shape.
}
} Stephane
}
} NB: "comfortable" is probably not the right word, because your particle
will still look like a small spot on the screen.
}
}
}
} ----- Original Message ----
} From: "dbagrov-at-gmail.com" {dbagrov-at-gmail.com}
} To: nizets2-at-yahoo.com
} Sent: Fri, February 5, 2010 12:19:55 PM
} Subject: [Microscopy] TEM - a question about sampling and magnification
}
}
}
}
}
----------------------------------------------------------------------------
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} To  Subscribe/Unsubscribe --
http://www.microscopy.com/MicroscopyListserver
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}
----------------------------------------------------------------------------
}
} Hello!
} I study nanoparticles by transmission electron microscopy. After
} examining several samples I have faced to two questions:
} 1. What is the minimum magnification which is required to notice a
nanoparticle?
} 2. What is the minimum magnification which is required to describe the
} shape of a nanoparticle?
} The answer to the first question seems to depend on the particle size,
} the resolution (number of pixels) of CCD camera, and probably the
} physical size of CCD (according to the Nyquist theorem). Is there a
} formula to calculate the minimum required magnification?
} The second question seems more tricky than the first one (at least for
} me). To my mind, there should be at least two formulas (one for the
} minimum size and the other one for the maximum size of a particle).
} Could you please help me with these question? I was trying to find
} answers on the Internet, but I couldn't find any concise examples,
} neither the formulas.
} Thanks!
} --
} Sincerely yours,
} Dmitry Bagrov
}
} ==============================Original
Headers==============================
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} 1, 32 -- Message-ID:
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} 1, 32 -- Subject: TEM - a question about sampling and magnification
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} 1, 32 -- To: Microscopy-at-microscopy.com
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3, 37 -- Subject: Re: [Microscopy] TEM - a question about sampling and
magnification
3, 37 -- From: Dmitry Bagrov {dbagrov-at-gmail.com}
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20, 22 -- Subject: RE: [Microscopy] Re: TEM - a question about sampling and magnification
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From: wesaia-at-iastate.edu
Date: Fri, 5 Feb 2010 17:12:14 -0600
Subject: [Microscopy] Re: TEM - a question about sampling and magnification

Contents Retrieved from Microscopy Listserver Archives
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Daniel,

Pat McCurdy told me this is an FESEM and your resolution was terrific. The
51nm size should be no problem to see, although given the microscope, you
could probably try viewing in the 1-5 kV range, just to see if anything is
there.

Yes, the sucrose could be evaporated or decomposed by the beam (if the
current was high enough), but I believe you would see it bubbling in the
live image. I think the particles may be "getting lost" on the way to the
sample stub.

Your first reaction I think was right. Seeing 51nm particles should be a
walk in the park, even with a W gun on most instruments! Maybe the
particles have gone to the same place that all the single socks go to. Let
us all know if you find that place. We've all lost a lot of socks over the
years.

Ken Converse

owner


QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz

-----Original Message-----
X-from: daniel thayer [mailto:kanawai-at-hotmail.com]
Sent: Friday, February 05, 2010 11:11 AM
To: kenconverse-at-qualityimages.biz

You are headed in the right direction although your steps are mildly confusing.

As far as the number of pixels required to determine "shape", just consider what an extremely pixilated image would look like. Something 5 pixels long would look very blocky. You might be able to get a sense of it being either an elongated particle or round, but you won't be able to tell much more.

As Philip said it, you may wish to rephrase the problem as the level of detail that you want to see on the feature. That could be texture or shape. Then, convert that dimension into pixel size. Nyquist would say you need to oversample by two-fold. Practically, you want something that you can see and only a two-fold oversampling will leave you wondering.

Somewhere along the line, you will need to bring in the practical resolution of your scope. What can you really achieve compared to pixel size? Increasing the magnification will reduce the pixel dimensions, but once your pixels are on the order of the resolution, then you have crossed over into the realm of empty magnification. More pixels in the feature will not be helping you.

I hope this helps,
Warren

-----Original Message-----
X-from: dbagrov-at-gmail.com [mailto:dbagrov-at-gmail.com]
Sent: Friday, February 05, 2010 6:52 AM
To: wesaia-at-iastate.edu

Dear Stephane,
Thanks for your quick reply!
Actually I was asking from a more "theoretical" point of view: the
required magnification doesn't depend ONLY on the particle size and
the size of the screen. One of the reasons is that the resolution of
the microscope is finite and increasing magnification doesn't always
allow to measure the size and analyze the shape of small particles.
Let's say that X is the number of pixels in the CCD camera (typically
1000 or 2000), L is the physical size of the sample which is projected
on the CCD camera. If we increase the magnification, L decreases
proportionally (the analyzed area becomes smaller).
According to the Nyquist theorem the minimum size of particle that can
be noticed is 2L/X. Let's assume that I need 5x5 pixels to describe
the shape of a particle (I am not sure if it is correct). So if the
magnification is constant (L is constant) I will be able to describe
the shape of a particle which is larger than 5L/X. If a microscope
provides a 300x300 nm image with 500'000 magnification and I have a
1000x1000 pixel camera I will be able to describe the shape of
particles which are larger than 5x300 nm/1000=1,5 nm (if the
resolution of the microscope allows me to do so).
Is there a way to make these or similar estimations in general? How
many pixels is needed to describe the shape of a particle?
--
Sincerely yours,
Dmitry Bagrov

2010/2/5 Stephane Nizet {nizets2-at-yahoo.com} :
} The only formula you need is the multiplication sign.
} Let's say that 1mm is a "comfortable" size to notice a particle.
} if your particle is 10nm in size, you'll need a 100,000x mag to make it appear 1mm big on your screen.
} (1 mm=1000 µm=1000,000 nm)
} Knowing that, I think you can guess yourself at what apparent size you are able to describe a shape.
}
} Stephane
}
} NB: "comfortable" is probably not the right word, because your particle will still look like a small spot on the screen.
}
}
}
} ----- Original Message ----
} From: "dbagrov-at-gmail.com" {dbagrov-at-gmail.com}
} To: nizets2-at-yahoo.com
} Sent: Fri, February 5, 2010 12:19:55 PM
} Subject: [Microscopy] TEM - a question about sampling and magnification
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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}
} Hello!
} I study nanoparticles by transmission electron microscopy. After
} examining several samples I have faced to two questions:
} 1. What is the minimum magnification which is required to notice a nanoparticle?
} 2. What is the minimum magnification which is required to describe the
} shape of a nanoparticle?
} The answer to the first question seems to depend on the particle size,
} the resolution (number of pixels) of CCD camera, and probably the
} physical size of CCD (according to the Nyquist theorem). Is there a
} formula to calculate the minimum required magnification?
} The second question seems more tricky than the first one (at least for
} me). To my mind, there should be at least two formulas (one for the
} minimum size and the other one for the maximum size of a particle).
} Could you please help me with these question? I was trying to find
} answers on the Internet, but I couldn't find any concise examples,
} neither the formulas.
} Thanks!
} --
} Sincerely yours,
} Dmitry Bagrov
}
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From: Amanda.Apilado-at-yahoo.com
Date: Mon, 8 Feb 2010 06:51:10 -0600
Subject: [Microscopy] viaWWW: Thermal pad properties

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Email: Amanda.Apilado-at-yahoo.com
Name: Amanda Apilado

Organization: Hitachi

Title-Subject: [Filtered] Thermal pad properties

Message: We're currently evaluating our existing thermal pads being
used for ion milling process (they say it's a graphite thermal pad).
And some contamination issues on slider devices(after ion mill)are
suspected to be caused by defective thermal pads. Can someone help us
better understand how do these pads work during actual ion mill
process? And what are the recommended performance/mechanical tests
that can be utilized in order to qualify future supplies?
Thanks!

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From: oshel1pe-at-cmich.edu
Date: Mon, 8 Feb 2010 07:33:14 -0600
Subject: [Microscopy] Fwd: Ask-A-Microscopist

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Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
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} Date: Fri, 5 Feb 2010 16:26:01 -0800 (PST)
} From: Cassie Rosinski {cassie-at-cassierosinski.com}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Friday, February 05,
} 2010 at 04:26:00 PM.
}
} realname - Cassie Rosinski
} Email - cassie-at-cassierosinski.com
} EDUCATION - Undergraduate College
} LOCATION - Portland OR USA
} SUBJECT_OF_QUESTION - EM programs
} QUESTION - Hi,
}
} I am currently an undergraduate chemistry major with 2 more years
} until graduation. I am strongly considering graduate school, and am
} very interested in learning more about EM as a whole, and exploring
} possible career paths.
}
} I would appreciate any information someone could give me about
} possible graduate (or undergraduate) programs in which I could
} inquire about studies. The only information I've been able to
} gather is on a few EM technician programs at 2 year colleges,
} however, I am more interested in graduate programs.
}
} Thanks in advance for any help you could give me in pointing me in
} the right direction.
}
} Cassie Rosinski
}

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From: joelsheffield-at-gmail.com
Date: Mon, 8 Feb 2010 09:23:13 -0600
Subject: [Microscopy] sucrose

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

You might do better to consider the old TEM process of shadowing. You could evaporate Pt from
a low angle, and try again to resolve the particles with the SEM. However, you may also be
getting very close to the resolving limit of your instrument. Do you know what the spot size is?

Joel


On 4 Feb 2010 at 18:28, jquinn-at-www.matscieng.sunysb.edu wrote:

}
}
}
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} Daniel and those "Out of Office"
}
} 15nm x 65,000 = 975,000nm = 975 microns = 0.975 mm
}
} Basically, your 15nm particle is a speck on the screen.
}
} Next, you are uniformaly coating the specks and substrate
} with gold, at 1/3 the speck size.
}
} It is not surprising that you do not see anything.
}
} Especially, since you are using 15kV on a light-element (sucrose)
} particle. The scattering volume dwarfs the speck size.
}
} just my two cents,
}
} JQuinn
}
} }
} } Date: Thu, 4 Feb 2010 13:30:43 -0800 (PST)
} } From: Daniel Thayer {kanawai-at-hotmail.com}
} } Subject: Ask-A-Microscopist
} }
} } Below is the result of your form, submitted on Thursday, February
} } 04, 2010 at 01:30:36 PM.
} }
} } realname - Daniel Thayer
} } Email - kanawai-at-hotmail.com
} } ORGANIZATION - Colorado State University
} } EDUCATION - Graduate College
} } LOCATION - Fort Collins, Colorado, United States
} } SUBJECT_OF_QUESTION - imaging sucrose particles by SEM
} } QUESTION - I am collecting sucrose particles on an aluminum plate
} } and imaging them with a SEM. I'm fairly new to the process and am
} } having trouble finding any particles. I calculated that there should
} } be about 23 particles per field at the concentrations generated for
} } the test. I am able to bring the image into sharp focus, so I should
} } be able to see any particles in the field of view. I'd like to rule
} } out the possiblility that the reason for this problem is that the
} } particles are evaporating under the electron beam when I try to
} } image them. The particles are 15 nm and I am imaging them at 65,000X
} } at 15.0 kV. The particles also contain trace amounts of ammonium
} } acetate from the buffer solution used while creating them. The
} } sample is sputter-coated with 5 nm of gold prior to imaging. Should
} } these conditions cause the particles to evaporate before I can focus
} } on them?
} }
} } Thanks,
} }
} } Daniel
} }
}
} ==============================Original Headers==============================
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} 9, 12 -- To: microscopy-at-microscopy.com
} 9, 12 -- Subject: re: sucrose
} ==============================End of - Headers==============================


--
Joel B. Sheffield, Ph.D.
Biology Department, Temple University
1900 North 12th Street
Philadelphia, PA 19122
jbs-at-temple.edu
(215) 204 8839, fax (215) 204 0486
http://astro.temple.edu/~jbs


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From: oshel1pe-at-cmich.edu
Date: Mon, 8 Feb 2010 13:30:31 -0600
Subject: [Microscopy] RE: Fwd: Ask-A-Microscopist microscopy courses

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

There is at least one 4-year undergrad program: ours. Central
Michigan U. offers a microscopy concentration within the B.Sc.
Biology major. The students take the usual biology courses (cell,
developmental, genetics, etc.), plus (separate) courses in Light
Microscopy, Confocal Light Microscopy, Scanning Electron Microscopy,
and Transmission Electron Microscopy, plus do an independent research
class (one or more semesters) using microscopy as the major research
tool. These courses are at the graduate level, but also available
(obviously) to undergrads. While this is a Biology sequence, we have
students from Chemistry and Physics taking these courses.
But, we do not have a M.Sc. or Ph.D. microscopy program, and I agree
with Norm and others -- it's better to find the research that
interests you, and just happens to use microscopy as a major tool.
Find a prof doing that, and you'll likely find graduate-level
microscopy courses.
Having said all that, don't discount the 2-year programs at San
Joaquin Delta Community College and Madison Area Technical College.
Associate degrees they may be, but I know more than one person who's
gotten a B.Sc. then gone through one of those programs -- and then
gotten a good job in industry or academics.

Phil

} Cassie: There are no EM programs in which I am aware that offer an
} advanced degree. The only ones that I know of are the ones you also
} found and they usually offer associates degrees. Microscopy is a
} tool used by many disciplines. I have individuals from Biology to
} Materials Science who use my instruments. Most graduate students
} who learn microscopy join a lab that does microscopy and the M.S. or
} Ph.D. degree is in that discipline not in microscopy.
}
} I guess what I would suggest is to see what area of chemistry you
} find to be the most compelling then start researching schools that
} have faculty members who work in those areas. Check out the faculty
} members' web sites and see how prominent microscopy is in their
} research. Microscopy is an extremely broad field. I am a member of
} the Microscopy Society of America but many times it seems like we
} all speak different languages and the only commonality is the tool
} we all use.
}
} Another possibility is to attend our annual meeting. There is an
} option where students, who don't mind working, get a major discount
} on the entrance fee. If you visit the exhibit room you will see
} more posters on various subjects than you could ever hope to keep in
} your mind.
}
} Good Luck
}
} Norm Olson
} University of California San Diego
} ________________________________________
} From: oshel1pe-at-cmich.edu [oshel1pe-at-cmich.edu]
} Sent: Monday, February 08, 2010 5:34 AM
} To: Olson, Norman
} Subject: [Microscopy] Fwd: Ask-A-Microscopist
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: mmiralles-at-pi.ac.ae
Date: Tue, 9 Feb 2010 05:36:06 -0600
Subject: [Microscopy] Low Carbon Steel Grain Boundaries

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

Hi everyone,

Someone from our ME department requested me to take a look on the grain boundaries of her low carbon steel sample. I haven't viewed carbon steel before but I have a metallography manual somewhere and from it I used the (10ml HCl + 5ml HNO3 + 85ml ethanol) etchant. Light repolishing was done after etching but I still couldn't get a clearer surface for image analysis. Suggestions?

Many thanks,

Melina Miralles
Petroleum Geosciences Lab Technician
The Petroleum Institute
Abu Dhabi, UAE
 



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From: Frank_Karl-at-lincolnelectric.com
Date: Tue, 9 Feb 2010 06:08:21 -0600
Subject: [Microscopy] Re: Low Carbon Steel Grain Boundaries

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

Hello Melina,
I would suggest you initially grind your sample though 120, 220, 600 and
1200 grit. Wash the sample and make sure there are not any wire edges or
burrs after the 1200. These could damage your fine polish. 1200 grit is
about 9um so then you want 6, 3 and 1 um diamond polish with careful
washing and cleaning between each step. Following the last diamond, wash,
rinse with methanol and hot air dry. I would try one of three different
etchants.

Picral, 4% picric acid in methanol (picric acid stains fingers and nails so
wear gloves) Also forms explosive compounds with many metals so lets keep
the express "Works a blast", just an expression.
Nital 4% nitric acid in methanol
Wheelers: 50/50 mix of picral and nital. It's kind of a mad dog etchant
and acts fast and is very easy to over etch with.

Picral attacks the interface between ferrite and carbide, nital attacks
ferrite grain and boundaries. And wheelers attacks both.

Don't over etch, what looks good to the eye is often over done. Over
etched samples can be repolished on the 3 and 1 micron diamond and then
re-etched.

Weld are easy to etch because they have structure different from base
metal, base metal takes practice. Have fun and let me know how it works
out......

Frank






mmiralles-at-pi.ac.a
e
To
02/09/2010 06:44 frank_karl-at-lincolnelectric.com
AM cc

Subject
Please respond to [Microscopy] Low Carbon Steel Grain
mmiralles-at-pi.ac.a Boundaries
e












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Hi everyone,

Someone from our ME department requested me to take a look on the grain
boundaries of her low carbon steel sample. I haven't viewed carbon steel
before but I have a metallography manual somewhere and from it I used the
(10ml HCl + 5ml HNO3 + 85ml ethanol) etchant. Light repolishing was done
after etching but I still couldn't get a clearer surface for image
analysis. Suggestions?

Many thanks,

Melina Miralles
Petroleum Geosciences Lab Technician
The Petroleum Institute
Abu Dhabi, UAE




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==============================Original Headers==============================
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From: richard.ross-at-allisontransmission.com
Date: Tue, 9 Feb 2010 07:29:22 -0600
Subject: [Microscopy] Re: Low Carbon Steel Grain Boundaries

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

Melina,
We review dozens of steel samples daily as a process control for our heat
treating operations (several processes - carburizing, nitriding, and
through-hardening of steels). Our routine procedure is to polish through 1
micron diamond and then etch for 5 seconds +/- with 5-10% HNO3 in
methanol.
Hope this helps,
Rick

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From: paul.gerroir-at-xrcc.xeroxlabs.com
Date: Tue, 9 Feb 2010 08:04:33 -0600
Subject: [Microscopy] SEM - JEOL JSM-6300F Donation

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

February 9, 2010

All,
We have for donation, a field emission gun (FEG)SEM purchased in 1992
and maintained on a service contract since that date. EDS: Kevex Delta
class (10mm2) detector with PGT analyzer (Spirit software).

We are willing to donate this instrument to any institution/organization
that accepts the dismantling and shipping charges. Installation will
also be the recipient's responsibility.

The microscope is being disassembled the week of February 14, 2010 so if
you are interested you must act quickly.

For more information contact me directly.

Paul

Paul J. Gerroir

Microscopy
Materials Characterization
Xerox Research Centre of Canada
2660 Speakman Drive
Mississauga, Ontario L5K 2L1

Phone: 905-823-7091, ext.216
FAX: 905-822-7022
e-mail: paul.gerroir-at-xerox.com



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From: hkonishi-at-wisc.edu
Date: Tue, 9 Feb 2010 12:58:18 -0600
Subject: [Microscopy] TEM sample of magnetic materials

Contents Retrieved from Microscopy Listserver Archives
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I would like to know how to prepare TEM samples of magnetic materials and how to evaluate if the TEM samples are not harmful to microscopes. Magnetic materials may be attracted by a pole piece. Therefore, they can damage the microscope. However, we know many people have characterized magnetic materials using TEM (e.g., bacterial magnetites and thin films). I also have experiences looking at maghemite particles from soil and magnetite inclusions in minerals. I was able to see the materials without any damage to the microscope. The materials could stay on the grids. It may depend on the particle size and the material how the magnetic materials are harmful to the microscope.

I would like to know:
1. How to prepare ion milled sample that is not harmful to scopes. I am afraid if ion milled samples are broken inside TEM and attracted by the pole piece.
2. Can I bring powder samples (magnetite) into microscopes?
3. How to make sure that the TEM samples are not harmful to microscopes?
4. Any other technique you recommend?

Thank you,

Hiromi Konishi, Ph.D.
UW _Madison

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From: ZZhang-at-uwyo.edu
Date: Tue, 9 Feb 2010 13:07:45 -0600
Subject: [Microscopy] Problem of TEM embedding of rat brain tissue

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

Hi All:

I have this 'weird' problem and need your help.

A student here has been trying do some TEM work with rat brain tissue. She first tried the "standard" protocol for embedding:

- Perfusion with 2% PFA, plus 2% GA
- Further fixation with 2% PFA, plus 2% GA, room temp, 1 hour
- 2nd fixation with 1% osmium, room temp, 1 hour
- Dehydration with EtoH (30....100%)
- Transition with propylene oxide
- Infiltration with 2:1, 1:2 (propylene oxide:resin), then 100% resin
- Polymerization (65C overnight)

The problem is that I always saw an incomplete infiltration (at least I thought so) - We were unable to cut even 2 um section. The sections fall apart, once it reaches the tissue. As I checked the semi-thin section, I saw holes all over the tissue.

I then asked the student tried to

1) extend the dehydration time (up to 3 min each step), and infiltration time (overnight each step)
2) additional microwave for infiltration;
3) smaller tissue (200 um vibritome section, instead of 1 mm cubes);
4) different resin (spur, embed812 and even LR white)

What is amazing to me is that NOTHING worked so far......

Any help?

Thank you.

Zhaojie Zhang, Ph.D.
Director, Microscopy Core Facility
Department of Zoology and Physiology
University of Wyoming
Laramie, WY 82071
zzhang-at-uwyo.edu
307-766-3038




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From: dsherman-at-purdue.edu
Date: Tue, 9 Feb 2010 13:19:21 -0600
Subject: [Microscopy] HPF of maize

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

Hi all,

Has anyone had success with high pressure freeze/freeze substitution of
maize coleoptile? We are interested in the vascular bundles and have
numerous problems.

The cells are really large with very little cytoplasm. If we cut the
samples really small we end up with lots of cut cells with nothing left
inside and often no vascular bundles. If the pieces are larger to increase
the chance of getting unbroken cells and vascular bundles than we get ice
damage. We had reasonable success with very young tissue but unfortunately
the investigators want a series of ages.If pieces are too small than
orientation becomes a real problem as well as tissue lose during processing.
Material is process for general morphology and also processed at low
temperature for ICC.

We get reasonable infiltration and contrast so that is not a problem. We
were thinking of possibly dicing the tissue in fixative (PAF or Karnovsky)
to just get a little more stability to the tissue and then, after about
5min, transferring to planchette filled with 0.15M sucrose and freezing. Has
anyone tried this? I know this partially defeats the reason for HPF but
getting poor tissue totally defeats it.

We also need to do Arabidopsis stem to get the vascular bundles and that
also gives us problems although not quite as bad as the coleoptile.

Debby


--
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy/



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From: DusevichV-at-umkc.edu
Date: Tue, 9 Feb 2010 13:23:39 -0600
Subject: [Microscopy] RE: Low Carbon Steel Grain Boundaries

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

Melina,
You are using too strong etchant. Already mentioned nital usually works
fine. Do not repolish after etching!
Good luck,
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

} -----Original Message-----
} From: mmiralles-at-pi.ac.ae [mailto:mmiralles-at-pi.ac.ae]
} Sent: Tuesday, February 09, 2010 5:37 AM
} To: Dusevich, Vladimir
} Subject: [Microscopy] Low Carbon Steel Grain Boundaries
}
}
}
}
}
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}
} Hi everyone,
}
} Someone from our ME department requested me to take a look on the
grain
} boundaries of her low carbon steel sample. I haven't viewed carbon
} steel before but I have a metallography manual somewhere and from it I
} used the (10ml HCl + 5ml HNO3 + 85ml ethanol) etchant. Light
} repolishing was done after etching but I still couldn't get a clearer
} surface for image analysis. Suggestions?
}
} Many thanks,
}
} Melina Miralles
} Petroleum Geosciences Lab Technician
} The Petroleum Institute
} Abu Dhabi, UAE
}
}
}
}
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} Headers==============================
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From: mcauliff-at-umdnj.edu
Date: Tue, 9 Feb 2010 13:24:15 -0600
Subject: [Microscopy] Re: Problem of TEM embedding of rat brain tissue

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

Was the perfusion successful? The tissue should be (mostly) clear of
blood and have a yellow tint due to glut. action.
How big were the pieces of tissue for subsequent processing? VERY important!
Low long was the dehydration in EtOH?
How long was the infiltration with 100% resin? How many changes?

You certainly should not be having problems after the changes you made.
The problem is with initial fixation, or lack there of.. Everything
hinges on good fixation.
Make fresh fix with ALL fresh components. Do it with the student present
so possible errors can be found.
Does the tissue blacken with osmium?
3 min. dehydration is just sufficient, even with 200 micron sections, so
that could be one of the areas of concern.
Fresh ethanols.
It does not matter how long poorly fixed tissue is in resin or what
resin is used.

Good luck!

Geoff

ZZhang-at-uwyo.edu wrote:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} ----------------------------------------------------------------------------
}
} Hi All:
}
} I have this 'weird' problem and need your help.
}
} A student here has been trying do some TEM work with rat brain tissue. She first tried the "standard" protocol for embedding:
}
} - Perfusion with 2% PFA, plus 2% GA
} - Further fixation with 2% PFA, plus 2% GA, room temp, 1 hour
} - 2nd fixation with 1% osmium, room temp, 1 hour
} - Dehydration with EtoH (30....100%)
} - Transition with propylene oxide
} - Infiltration with 2:1, 1:2 (propylene oxide:resin), then 100% resin
} - Polymerization (65C overnight)
}
} The problem is that I always saw an incomplete infiltration (at least I thought so) - We were unable to cut even 2 um section. The sections fall apart, once it reaches the tissue. As I checked the semi-thin section, I saw holes all over the tissue.
}
} I then asked the student tried to
}
} 1) extend the dehydration time (up to 3 min each step), and infiltration time (overnight each step)
} 2) additional microwave for infiltration;
} 3) smaller tissue (200 um vibritome section, instead of 1 mm cubes);
} 4) different resin (spur, embed812 and even LR white)
}
} What is amazing to me is that NOTHING worked so far......
}
} Any help?
}
} Thank you.
}
} Zhaojie Zhang, Ph.D.
} Director, Microscopy Core Facility
} Department of Zoology and Physiology
} University of Wyoming
} Laramie, WY 82071
} zzhang-at-uwyo.edu
} 307-766-3038
}
}
}
}
} ==============================Original Headers==============================
} 14, 27 -- From ZZhang-at-uwyo.edu Tue Feb 9 13:07:44 2010
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} 14, 27 -- Date: Tue, 9 Feb 2010 12:07:39 -0700
} 14, 27 -- Subject: Problem of TEM embedding of rat brain tissue
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--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff-at-umdnj.edu
**********************************************



==============================Original Headers==============================
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9, 28 -- To: ZZhang-at-uwyo.edu, microscopy-at-microscopy.com
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9, 28 -- References: {201002091908.o19J8RYn025363-at-ns.microscopy.com}
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==============================End of - Headers==============================




From: ZZhang-at-uwyo.edu
Date: Tue, 9 Feb 2010 14:58:37 -0600
Subject: [Microscopy] Correction - Problem of TEM embedding of rat brain tissue

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

I apologize for the typo in my original posting!

The original dehydration is 15 min each step, then extended to 30 min (not 3 min) each step.
The size of the vibratome section is 200 um thick, then 2X2 mm.

Thank you,

Zhaojie

} Hi All:
}
} I have this 'weird' problem and need your help.
}
} A student here has been trying do some TEM work with rat brain tissue. She first tried the "standard" protocol for embedding:
}
} - Perfusion with 2% PFA, plus 2% GA
} - Further fixation with 2% PFA, plus 2% GA, room temp, 1 hour
} - 2nd fixation with 1% osmium, room temp, 1 hour
} - Dehydration with EtoH (30....100%)
} - Transition with propylene oxide
} - Infiltration with 2:1, 1:2 (propylene oxide:resin), then 100% resin
} - Polymerization (65C overnight)
}
} The problem is that I always saw an incomplete infiltration (at least I thought so) - We were unable to cut even 2 um section. The sections fall apart, once it reaches the tissue. As I checked the semi-thin section, I saw holes all over the tissue.
}
} I then asked the student tried to
}
} 1) extend the dehydration time (up to 3 min each step), and infiltration time (overnight each step)
} 2) additional microwave for infiltration;
} 3) smaller tissue (200 um vibritome section, instead of 1 mm cubes);
} 4) different resin (spur, embed812 and even LR white)
}
} What is amazing to me is that NOTHING worked so far......
}
} Any help?
}
} Thank you.
}
} Zhaojie Zhang, Ph.D.
} Director, Microscopy Core Facility
} Department of Zoology and Physiology
} University of Wyoming
} Laramie, WY 82071
} zzhang-at-uwyo.edu
} 307-766-3038
}
}
}
}
} ==============================Original Headers==============================
} 14, 27 -- From ZZhang-at-uwyo.edu Tue Feb 9 13:07:44 2010
} 14, 27 -- Received: from aspensprings.uwyo.edu (aspensprings.uwyo.edu [129.72.10.32])
} 14, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id o19J7gE4023595
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} 14, 27 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
} 14, 27 -- Date: Tue, 9 Feb 2010 12:07:39 -0700
} 14, 27 -- Subject: Problem of TEM embedding of rat brain tissue
} 14, 27 -- Thread-Topic: Problem of TEM embedding of rat brain tissue
} 14, 27 -- Thread-Index: Acqpuc0JQft/GEA0SBK0wrHyjH+BDQAAUEfQ
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--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff-at-umdnj.edu
**********************************************



==============================Original Headers==============================
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9, 28 -- To: ZZhang-at-uwyo.edu, microscopy-at-microscopy.com
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9, 28 -- References: {201002091908.o19J8RYn025363-at-ns.microscopy.com}
9, 28 -- In-reply-to: {201002091908.o19J8RYn025363-at-ns.microscopy.com}
==============================End of - Headers==============================


==============================Original Headers==============================
10, 29 -- From ZZhang-at-uwyo.edu Tue Feb 9 14:58:37 2010
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10, 29 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
10, 29 -- Date: Tue, 9 Feb 2010 13:58:30 -0700
10, 29 -- Subject: Correction - Problem of TEM embedding of rat brain tissue
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From: DusevichV-at-umkc.edu
Date: Tue, 9 Feb 2010 15:38:24 -0600
Subject: [Microscopy] Problem of TEM embedding of rat brain tissue

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

Geoff,
How can fixation influence infiltration?
I suspect ethanol. Is it really 100%? Was it stored for more or less
long time in opened bottle?
Also, how many changes of pure resin?
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784


} -----Original Message-----
} From: mcauliff-at-umdnj.edu [mailto:mcauliff-at-umdnj.edu]
} Sent: Tuesday, February 09, 2010 1:25 PM
} To: Dusevich, Vladimir
} Subject: [Microscopy] Re: Problem of TEM embedding of rat brain tissue
}
}
}
}
}
-----------------------------------------------------------------------
} -----
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
}
-----------------------------------------------------------------------
} -----
}
} Was the perfusion successful? The tissue should be (mostly) clear of
} blood and have a yellow tint due to glut. action.
} How big were the pieces of tissue for subsequent processing? VERY
} important!
} Low long was the dehydration in EtOH?
} How long was the infiltration with 100% resin? How many changes?
}
} You certainly should not be having problems after the changes you
made.
} The problem is with initial fixation, or lack there of.. Everything
} hinges on good fixation.
} Make fresh fix with ALL fresh components. Do it with the student
} present
} so possible errors can be found.
} Does the tissue blacken with osmium?
} 3 min. dehydration is just sufficient, even with 200 micron sections,
} so
} that could be one of the areas of concern.
} Fresh ethanols.
} It does not matter how long poorly fixed tissue is in resin or what
} resin is used.
}
} Good luck!
}
} Geoff
}
} ZZhang-at-uwyo.edu wrote:
} }
---------------------------------------------------------------------
} -------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} } To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} }
---------------------------------------------------------------------
} -------
} }
} } Hi All:
} }
} } I have this 'weird' problem and need your help.
} }
} } A student here has been trying do some TEM work with rat brain
} tissue. She first tried the "standard" protocol for embedding:
} }
} } - Perfusion with 2% PFA, plus 2% GA
} } - Further fixation with 2% PFA, plus 2% GA, room temp, 1 hour
} } - 2nd fixation with 1% osmium, room temp, 1 hour
} } - Dehydration with EtoH (30....100%)
} } - Transition with propylene oxide
} } - Infiltration with 2:1, 1:2 (propylene oxide:resin), then 100%
resin
} } - Polymerization (65C overnight)
} }
} } The problem is that I always saw an incomplete infiltration (at
least
} I thought so) - We were unable to cut even 2 um section. The sections
} fall apart, once it reaches the tissue. As I checked the semi-thin
} section, I saw holes all over the tissue.
} }
} } I then asked the student tried to
} }
} } 1) extend the dehydration time (up to 3 min each step), and
} infiltration time (overnight each step)
} } 2) additional microwave for infiltration;
} } 3) smaller tissue (200 um vibritome section, instead of 1 mm cubes);
} } 4) different resin (spur, embed812 and even LR white)
} }
} } What is amazing to me is that NOTHING worked so far......
} }
} } Any help?
} }
} } Thank you.
} }
} } Zhaojie Zhang, Ph.D.
} } Director, Microscopy Core Facility
} } Department of Zoology and Physiology
} } University of Wyoming
} } Laramie, WY 82071
} } zzhang-at-uwyo.edu
} } 307-766-3038
} }
} }
} }
} }
} } ==============================Original
} Headers==============================
} } 14, 27 -- From ZZhang-at-uwyo.edu Tue Feb 9 13:07:44 2010
} } 14, 27 -- Received: from aspensprings.uwyo.edu
(aspensprings.uwyo.edu
} [129.72.10.32])
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} } 14, 27 -- Date: Tue, 9 Feb 2010 12:07:39 -0700
} } 14, 27 -- Subject: Problem of TEM embedding of rat brain tissue
} } 14, 27 -- Thread-Topic: Problem of TEM embedding of rat brain tissue
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} }
} }
} }
}
}
} --
} --
} **********************************************
} Geoff McAuliffe, Ph.D.
} Neuroscience and Cell Biology
} Robert Wood Johnson Medical School
} 675 Hoes Lane, Piscataway, NJ 08854
} voice: (732)-235-4583
} mcauliff-at-umdnj.edu
} **********************************************
}
}
}
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} brain tissue
} 9, 28 -- References: {201002091908.o19J8RYn025363-at-ns.microscopy.com}
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6, 25 -- From DusevichV-at-umkc.edu Tue Feb 9 15:38:23 2010
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From: ZZhang-at-uwyo.edu
Date: Tue, 9 Feb 2010 15:49:59 -0600
Subject: [Microscopy] Problem of TEM embedding of rat brain tissue

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

That was a good question. I never thought that fixation would affect infiltration, yet several people suggested so. Does anyone know why?

As for ethanol, we always use a freshly opened bottle for 100% with 3 changes.
For pure resin, we make at least 2 changes (total 12 hours infiltration in 100% resin)

Zhaojie

-----Original Message-----
X-from: DusevichV-at-umkc.edu [mailto:DusevichV-at-umkc.edu]
Sent: Tuesday, February 09, 2010 2:44 PM
To: Z.J. Zhang

Geoff,
How can fixation influence infiltration?
I suspect ethanol. Is it really 100%? Was it stored for more or less
long time in opened bottle?
Also, how many changes of pure resin?
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784


} -----Original Message-----
} From: mcauliff-at-umdnj.edu [mailto:mcauliff-at-umdnj.edu]
} Sent: Tuesday, February 09, 2010 1:25 PM
} To: Dusevich, Vladimir
} Subject: [Microscopy] Re: Problem of TEM embedding of rat brain tissue
}
}
}
}
}
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} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
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}
} Was the perfusion successful? The tissue should be (mostly) clear of
} blood and have a yellow tint due to glut. action.
} How big were the pieces of tissue for subsequent processing? VERY
} important!
} Low long was the dehydration in EtOH?
} How long was the infiltration with 100% resin? How many changes?
}
} You certainly should not be having problems after the changes you
made.
} The problem is with initial fixation, or lack there of.. Everything
} hinges on good fixation.
} Make fresh fix with ALL fresh components. Do it with the student
} present
} so possible errors can be found.
} Does the tissue blacken with osmium?
} 3 min. dehydration is just sufficient, even with 200 micron sections,
} so
} that could be one of the areas of concern.
} Fresh ethanols.
} It does not matter how long poorly fixed tissue is in resin or what
} resin is used.
}
} Good luck!
}
} Geoff
}
} ZZhang-at-uwyo.edu wrote:
} }
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} -------
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} -------
} }
} } Hi All:
} }
} } I have this 'weird' problem and need your help.
} }
} } A student here has been trying do some TEM work with rat brain
} tissue. She first tried the "standard" protocol for embedding:
} }
} } - Perfusion with 2% PFA, plus 2% GA
} } - Further fixation with 2% PFA, plus 2% GA, room temp, 1 hour
} } - 2nd fixation with 1% osmium, room temp, 1 hour
} } - Dehydration with EtoH (30....100%)
} } - Transition with propylene oxide
} } - Infiltration with 2:1, 1:2 (propylene oxide:resin), then 100%
resin
} } - Polymerization (65C overnight)
} }
} } The problem is that I always saw an incomplete infiltration (at
least
} I thought so) - We were unable to cut even 2 um section. The sections
} fall apart, once it reaches the tissue. As I checked the semi-thin
} section, I saw holes all over the tissue.
} }
} } I then asked the student tried to
} }
} } 1) extend the dehydration time (up to 3 min each step), and
} infiltration time (overnight each step)
} } 2) additional microwave for infiltration;
} } 3) smaller tissue (200 um vibritome section, instead of 1 mm cubes);
} } 4) different resin (spur, embed812 and even LR white)
} }
} } What is amazing to me is that NOTHING worked so far......
} }
} } Any help?
} }
} } Thank you.
} }
} } Zhaojie Zhang, Ph.D.
} } Director, Microscopy Core Facility
} } Department of Zoology and Physiology
} } University of Wyoming
} } Laramie, WY 82071
} } zzhang-at-uwyo.edu
} } 307-766-3038
} }
} }
} }
} }
} } ==============================Original
} Headers==============================
} } 14, 27 -- From ZZhang-at-uwyo.edu Tue Feb 9 13:07:44 2010
} } 14, 27 -- Received: from aspensprings.uwyo.edu
(aspensprings.uwyo.edu
} [129.72.10.32])
} } 14, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with
} ESMTP id o19J7gE4023595
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} ht2.uwyo.edu [10.84.60.209])
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} } 14, 27 -- From: "Z.J. Zhang" {ZZhang-at-uwyo.edu}
} } 14, 27 -- To: "microscopy-at-microscopy.com"
{microscopy-at-microscopy.com}
} } 14, 27 -- Date: Tue, 9 Feb 2010 12:07:39 -0700
} } 14, 27 -- Subject: Problem of TEM embedding of rat brain tissue
} } 14, 27 -- Thread-Topic: Problem of TEM embedding of rat brain tissue
} } 14, 27 -- Thread-Index: Acqpuc0JQft/GEA0SBK0wrHyjH+BDQAAUEfQ
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} }
} }
}
}
} --
} --
} **********************************************
} Geoff McAuliffe, Ph.D.
} Neuroscience and Cell Biology
} Robert Wood Johnson Medical School
} 675 Hoes Lane, Piscataway, NJ 08854
} voice: (732)-235-4583
} mcauliff-at-umdnj.edu
} **********************************************
}
}
}
} ==============================Original
} Headers==============================
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} 9, 28 -- To: ZZhang-at-uwyo.edu, microscopy-at-microscopy.com
} 9, 28 -- Subject: Re: [Microscopy] Problem of TEM embedding of rat
} brain tissue
} 9, 28 -- References: {201002091908.o19J8RYn025363-at-ns.microscopy.com}
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From: matthew.weyland-at-mcem.monash.edu.au
Date: Tue, 9 Feb 2010 16:02:52 -0600
Subject: [Microscopy] Re: TEM sample of magnetic materials

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

Dear Hiromi,

1. As you say, many people study bulk magnetic specimens inside the TEM, indeed
some microscopists use magnetic materials as support grids (such as Ni). As long
as your sample holder is a design which rigidly holds the specimen in place
(such as a hex nut, rather than a plate and/or arm clamps) the bulk of the
specimen will not be pulled out of the holder! As for thin regions being torn
away, there is some possibility of this occurring - but I am pretty sure the
risk is small (those who work on bulk magnetics, please correct me!). It will of
course depend on the mechanical properties of the material in question, with
brittle materials being more of a problem.

2. Powdered magnetic samples are not a problem, as long as they are suitably
small for TEM study ( {100 nm). Van der Waals forces are MUCH more powerful on
these length scales than magnetic moments and wouldn't detach from your grid.

3. I'm not sure what you mean by harmful? What you will experience with bulk
magnetics is a misalignment of the instrument every time the specimen is tilted
or moved a large distance. Simply as the specimen acts on the electrons in the
beam and interacts with the field of the lens. I would recommend the lowest
resolution instrument you can find for bulk magnetics, as the wider the
polepiece gap the smaller this effect will be. And don't even think about an
aberration corrected microscope! None of these misalignments will be permanent
so no 'harm' will occur.

If you managed to tear a specimen out of a holder (unlikely, see point 1) it
might potentially lodge in a awkward place on the pole piece blocking the beam
or apply an aberration to the lens. In this situation powering down and venting
the pole piece would be required to remove the specimen. Any thin
area/nanoparticles removed onto the polepiece will have an inconsequential
effect on the microsope, their volume is simply too small.

4. If you are really concerned about operating in a field you can use a
microscope with a Lorentz lens - turning off the objective and working in a
field free environment at the specimen. However, with such far field focusing
you will lose a large portion of your resolution.

Regards

Matthew

--
Dr M.Weyland, Senior Research Fellow & Electron Microscopist
----------------------------------------------------------------------------
|Monash Centre for Electron Microscopy |Department of Materials Engineering|
|Bldg 81 |Room 115, Bldg 69 |
|Monash University |Monash University |
|Victoria 3800 |Victoria 3800 |
|Australia. |Australia |
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hkonishi-at-wisc.edu wrote:
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}
} I would like to know how to prepare TEM samples of magnetic materials and how to evaluate if the TEM samples are not harmful to microscopes. Magnetic materials may be attracted by a pole piece. Therefore, they can damage the microscope. However, we know many people have characterized magnetic materials using TEM (e.g., bacterial magnetites and thin films). I also have experiences looking at maghemite particles from soil and magnetite inclusions in minerals. I was able to see the materials without any damage to the microscope. The materials could stay on the grids. It may depend on the particle size and the material how the magnetic materials are harmful to the microscope.
}
} I would like to know:
} 1. How to prepare ion milled sample that is not harmful to scopes. I am afraid if ion milled samples are broken inside TEM and attracted by the pole piece.
} 2. Can I bring powder samples (magnetite) into microscopes?
} 3. How to make sure that the TEM samples are not harmful to microscopes?
} 4. Any other technique you recommend?
}
} Thank you,
}
} Hiromi Konishi, Ph.D.
} UW _Madison
}


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From: rosemary.white-at-csiro.au
Date: Tue, 9 Feb 2010 16:50:17 -0600
Subject: [Microscopy] Re: HPF of maize

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

Hi Debby,

Is there a particular reason you're trying HPF here? Maize coleoptiles are
(relatively) large, cells are vacuolate and the tissue contains quite a lot
of air so they would seem rather poor candidates for HPF. If you have to
freeze, why not try plunge freezing - good methods can be found in papers of
Peter Hepler or Tobias Baskin. With such big cells, ice damage is
inevitable, I'm guessing. The vascular tissue is too deeply buried in the
coleoptile to be preserved quickly enough to avoid damage.

Maize coleoptiles usually have only two vascular bundles on opposite sides
of the coleoptile, though there can sometimes be more. This might explain
why some pieces have no vascular tissue.

Standard fixation for immuno work - paraformaldehyde in buffer of choice
with or without a little glut and other additives (Mg, Ca, etc.) gives good
results for tissue embedded in LRWhite, if that's what you're after. To
track tissue, add a very little fast green (if dehydrated in alcohol) or
alizarin red (if dehydrated in acetone) so you can still see the tissue in
liquid resin.

We've done quite a bit of immuno work on freshly fixed tissue, as well as
embedded, and heaps on Arabidopsis - unless there's something soluble you
really want to pin down that is redistributed during processing,
conventional fixation works well.

cheers,
Rosemary

Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
M 61 2 420 972 028


On 10/02/10 6:25 AM, "dsherman-at-purdue.edu" {dsherman-at-purdue.edu} wrote:

}
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} Hi all,
}
} Has anyone had success with high pressure freeze/freeze substitution of
} maize coleoptile? We are interested in the vascular bundles and have
} numerous problems.
}
} The cells are really large with very little cytoplasm. If we cut the
} samples really small we end up with lots of cut cells with nothing left
} inside and often no vascular bundles. If the pieces are larger to increase
} the chance of getting unbroken cells and vascular bundles than we get ice
} damage. We had reasonable success with very young tissue but unfortunately
} the investigators want a series of ages.If pieces are too small than
} orientation becomes a real problem as well as tissue lose during processing.
} Material is process for general morphology and also processed at low
} temperature for ICC.
}
} We get reasonable infiltration and contrast so that is not a problem. We
} were thinking of possibly dicing the tissue in fixative (PAF or Karnovsky)
} to just get a little more stability to the tissue and then, after about
} 5min, transferring to planchette filled with 0.15M sucrose and freezing. Has
} anyone tried this? I know this partially defeats the reason for HPF but
} getting poor tissue totally defeats it.
}
} We also need to do Arabidopsis stem to get the vascular bundles and that
} also gives us problems although not quite as bad as the coleoptile.
}
} Debby
}



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From: ZZhang-at-uwyo.edu
Date: Tue, 9 Feb 2010 17:13:30 -0600
Subject: [Microscopy] RE: Problem of TEM embedding of rat brain tissue

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

Then the washed-out area would be filled with resin, causing a structure problem, but not infiltration problem, right?

-----Original Message-----
X-from: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu]
Sent: Tuesday, February 09, 2010 3:10 PM
To: Z.J. Zhang


--
Lee Cohen-Gould, M.S.
Sr. Staff Associate in Biochemistry and
Cell & Developmental Biology
Director, Electron Microscopy & Histology
and Optical Microscopy Core Facilities
Weill Cornell Medical College

voice (212)746-6146
fax (212)746-8175
http://www.med.cornell.edu/research/rea_sup/
http://www.cornellcelldevbiology.org
http://www.cornellbiochem.org


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From: vladislav_speransky-at-nih.gov
Date: Tue, 9 Feb 2010 18:17:37 -0600
Subject: [Microscopy] Re: Problem of TEM embedding of rat brain tissue

Contents Retrieved from Microscopy Listserver Archives
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We have an old American Optical Ultracut waiting on the sidelines in
case our current ultramicrotome goes belly-up. I've never used an AO or
one of its newer incarnations before, but the only set of instructions
we have is a short form of the manual. Would anyone happen to have a
more complete set of documents for it? Or know where I could obtain
them? (PDFs would be sufficient.)

Thank you so much,

Jaclynn Lett
Senior Research Technician, EM Facility
Research Center for Auditory and Vestibular Studies
Department of Otolaryngology
Washington University School of Medicine
660 S. Euclid Ave., Campus Box 8115
St. Louis, MO 63110

Office: 314-747-7257
Fax: 314-747-7230
Email: lettj-at-ent.wustl.edu
http://otocore.wustl.edu




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From watches.watches-at-barb.com Tue Feb 9 17:55:27 2010
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Make it x3... :-)

Perhaps something Geoff has observed empirically?.. If I observed such
thing, the explanation I would offer would be:
One of the things an aldehyde fixative does is that it makes the
membranes permeable. This could be critical for efficient dehydration
and infiltration. Now that I think about it - that is indeed a more
widely known reason to why we fix in aldehyde to do Tokuyasu immunoEM:
to facilitate infusion with sucrose before freezing. Very likely the
same thing with resin embedding.

Otherwise, the pieces described are definitely thin enough, and
dehydration times are more than adequate, especially with PO.
Interesting.

Vlad
________________________________________________
Vlad Speransky, Staff Scientist
Supramolecular Structure and Function Resource
National Institute of Biomedical Imaging and Bioengineering, NIH
13 South Dr, Rm. 3N17 MSC 5766
Bethesda, MD 20892
301 496-3989
vladislav_speransky-at-nih.gov
http://www.nibib.nih.gov/Research/Intramural/lbps/staff/speransky

Opinions and experiences related are those of Vlad Speransky and do
not represent the NIH. On the good side, this message is not
confidential and can be freely shared and reproduced.


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From: mmiralles-at-pi.ac.ae
Date: Tue, 9 Feb 2010 23:14:07 -0600
Subject: [Microscopy] RE: Low Carbon Steel Grain Boundaries

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

Hi all,

As always, I have received favorable amount of responses that will keep
me busy (and quiet, for now!).

Thank you so much.

Regards,
Melina Miralles




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From: protrain-at-emcourses.com
Date: Wed, 10 Feb 2010 04:36:01 -0600
Subject: [Microscopy] Re: TEM sample of magnetic materials

Contents Retrieved from Microscopy Listserver Archives
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Hi

You have already had some good advice but may I add a little more?

Whilst it is conventional to operate a microscope with the specimen stage in
the eucentric position it is not essential to do so! With your magnetic
materials the lower the lens strength the lower the level of problems as
have been pointed out.

1. The "higher resolution" option is to adjust the eucentric stage so
that the specimen is moved away from the lower objective pole piece which
would mean a lower lens current/magnetic field during operation. Simply
adjust the Z prime (eucentric adjustment) so that the specimen remains in
focus as you turn your focus controls anticlockwise; for a lower lens
strength. You will probably obtain a focal length up to 2mm longer than
normal. This drop in objective lens current will require you to re
calibrate the magnification system.

2. The "low resolution" option is to run in a "low magnification" or
"scan" mode, when the objective lens is either switched off or run at a very
low current, in this case focus is achieved with the diffraction lens.

Point 1 is also a good idea for biologists struggling for contrast as it
increases the specimen to objective aperture distance, hence contrast. It
is also a method for obtaining "higher resolution" low magnification images
as the magnification range is likely to drop by a factor of up to 40%
depending upon the way the manufacturer uses their imaging lens system.

Enjoy!


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com




-----Original Message-----
X-from: matthew.weyland-at-mcem.monash.edu.au
[mailto:matthew.weyland-at-mcem.monash.edu.au]
Sent: 09 February 2010 22:04
To: protrain-at-emcourses.com

Dear Hiromi,

1. As you say, many people study bulk magnetic specimens inside the TEM,
indeed
some microscopists use magnetic materials as support grids (such as Ni). As
long
as your sample holder is a design which rigidly holds the specimen in place
(such as a hex nut, rather than a plate and/or arm clamps) the bulk of the
specimen will not be pulled out of the holder! As for thin regions being
torn
away, there is some possibility of this occurring - but I am pretty sure the

risk is small (those who work on bulk magnetics, please correct me!). It
will of
course depend on the mechanical properties of the material in question, with

brittle materials being more of a problem.

2. Powdered magnetic samples are not a problem, as long as they are suitably

small for TEM study ( {100 nm). Van der Waals forces are MUCH more powerful
on
these length scales than magnetic moments and wouldn't detach from your
grid.

3. I'm not sure what you mean by harmful? What you will experience with bulk

magnetics is a misalignment of the instrument every time the specimen is
tilted
or moved a large distance. Simply as the specimen acts on the electrons in
the
beam and interacts with the field of the lens. I would recommend the lowest
resolution instrument you can find for bulk magnetics, as the wider the
polepiece gap the smaller this effect will be. And don't even think about an

aberration corrected microscope! None of these misalignments will be
permanent
so no 'harm' will occur.

If you managed to tear a specimen out of a holder (unlikely, see point 1) it

might potentially lodge in a awkward place on the pole piece blocking the
beam
or apply an aberration to the lens. In this situation powering down and
venting
the pole piece would be required to remove the specimen. Any thin
area/nanoparticles removed onto the polepiece will have an inconsequential
effect on the microsope, their volume is simply too small.

4. If you are really concerned about operating in a field you can use a
microscope with a Lorentz lens - turning off the objective and working in a
field free environment at the specimen. However, with such far field
focusing
you will lose a large portion of your resolution.

Regards

Matthew

--
Dr M.Weyland, Senior Research Fellow & Electron Microscopist
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hkonishi-at-wisc.edu wrote:
}
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}
} I would like to know how to prepare TEM samples of magnetic materials and
how to evaluate if the TEM samples are not harmful to microscopes. Magnetic
materials may be attracted by a pole piece. Therefore, they can damage the
microscope. However, we know many people have characterized magnetic
materials using TEM (e.g., bacterial magnetites and thin films). I also
have experiences looking at maghemite particles from soil and magnetite
inclusions in minerals. I was able to see the materials without any damage
to the microscope. The materials could stay on the grids. It may depend on
the particle size and the material how the magnetic materials are harmful to
the microscope.
}
} I would like to know:
} 1. How to prepare ion milled sample that is not harmful to scopes. I am
afraid if ion milled samples are broken inside TEM and attracted by the pole
piece.
} 2. Can I bring powder samples (magnetite) into microscopes?
} 3. How to make sure that the TEM samples are not harmful to microscopes?
} 4. Any other technique you recommend?
}
} Thank you,
}
} Hiromi Konishi, Ph.D.
} UW _Madison
}


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From: LettJ-at-ent.wustl.edu
Date: Wed, 10 Feb 2010 10:44:28 -0600
Subject: [Microscopy] RMC MT-X ultramicrotome

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

We're looking for third-party service for the RMC MT-X ultramicrotome
(in particular, for the Midwest region). Does anyone have any leads for
us?

Thank you,

Jaci

Jaclynn Lett
Senior Research Technician, EM Facility
Research Center for Auditory and Vestibular Studies
Department of Otolaryngology
Washington University School of Medicine
660 S. Euclid Ave., Campus Box 8115
St. Louis, MO 63110

Office: 314-747-7257
Fax: 314-747-7230
Email: lettj-at-ent.wustl.edu
http://otocore.wustl.edu




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From: yaoz-at-me.queensu.ca
Date: Wed, 10 Feb 2010 15:37:00 -0600
Subject: [Microscopy] viaWWW: Requiring a used EDS detector

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Email: yaoz-at-me.queensu.ca
Name: yao

Organization: Queensu

Title-Subject: [Filtered] Requiring a used EDS detector

Message: We are looking for a used EDS detector to replace the
damaged one in our current EDS system. The model is: NORAN
7720B-4NES-SN NANOTRACE PHILIPS CM 20T.

If anybody is deposing of such one, please let me know the price of purchasing.

Thanks a lot!

Yao

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From: bschneid-at-fhcrc.org
Date: Wed, 10 Feb 2010 15:37:05 -0600
Subject: [Microscopy] viaWWW: Betty Mathews 1/12/27 -1/26/10

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Email: bschneid-at-fhcrc.org
Name: Bobbie Schneider

Organization: Fred Hutchinson Cancer Research Center

Title-Subject: [Filtered] Betty Mathews 1/12/27 -1/26/10

Message: Hello:

This message is to let the Delta Electron Microscopy Students know
that Dr. Mathews passed away on 1/26/10. Dr. Mathews was responsible
for creating the two year Electron Microscopy training program at San
Joaquin Delta College in Stockton, CA and was the mentor for many
technicians. Her memorial service will be held on Saturday, February
20, 2010 at the Blessed Sacrament Chapel, Carmel Mission, Carmel, CA
at 2:00PM. She will be greatly missed.

Bobbie Schneider
FHCRC
Seattle, WA 98109


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From: ryan.deacon-at-jhuapl.edu
Date: Wed, 10 Feb 2010 15:37:06 -0600
Subject: [Microscopy] viaWWW: Scroll pump for sputter coater

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Email: ryan.deacon-at-jhuapl.edu
Name: Ryan Deacon

Organization: Johns Hopkins University Applied Physics Lab

Title-Subject: [Filtered] Scroll pump for sputter coater

Message: Good morning list members -

We're in the process of replacing our sputter coater and are
considering the pros & cons of dry scroll pumps over traditional
rotary pumps for the new system. I'd be interested in hearing from
list members who have experience with scroll pumps on coating
systems: do they have a significant impact on reducing ample
contamination, are pump down times considerably longer, are they
noisy or difficult to maintain, etc.

The new system will be used primarily for Au/Pd or Ir coating of a
wide variety of samples for high res imaging on a cold FE SEM. My
current SEM has rotary pumps and a diffusion pump, but plans are for
future SEMs to have an oil-free vacuum system, so I am considering
impact on both my current and future SEM - we won't be getting
another coater for many years.

Thanks for your input.

Cheers - Ryan


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From: spratsbd-at-matthey.com
Date: Wed, 10 Feb 2010 15:37:13 -0600
Subject: [Microscopy] viaWWW: Fault Finding on a Precision Ion Polishing System

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Email: spratsbd-at-matthey.com
Name: Steve Spratt

Organization: Johnson Matthey plc.,

Title-Subject: [Filtered] Fault Finding on a Precision Ion Polishing System

Message: Have you a copy of a circuit and block diagram to trace and
identify a fault on our PIPS; Model 691?
Your assistance would be much appreciated.

Regards,

Steve

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From: patrick.mccurdy-at-colostate.edu
Date: Wed, 10 Feb 2010 15:37:15 -0600
Subject: [Microscopy] viaWWW: Stray fields spec versus reality

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Email: patrick.mccurdy-at-colostate.edu
Name: Patrick McCurdy

Organization: Colorado State University

Title-Subject: [Filtered] Stray fields spec versus reality

Message: We are having to move our SEM to make room for a new TEM. We
recently had a site survey done of our proposed site and it shows
that stray magnetic fields for both AC and DC exist and are above the
manufacture's requirements. Our scope is a JEOL6500F. Please email me
offline if you have experience with this situation. One option is to
put in a noise cancelation system, but this will interfere with other
instruments in the room and space is limited between instruments.

Sincerely,
Pat McCurdy
Colorado State University

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From: gary-at-gaugler.com
Date: Wed, 10 Feb 2010 18:28:33 -0600
Subject: [Microscopy] Re: viaWWW: Scroll pump for sputter coater

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

I have a Denton Desk IV TSC which is the prior turbo
model to the Desk V. I use an Edwards XDS-10 dry
scroll pump (external of course). I coat using Pd
or Ir, depending on the specimen and EDS conflicts.

I would say that the Edwards is about the same noise
as a rotary vane pump. However, the Desk uses a
diaphragm oil pump as their standard option and it
is quieter--but oil. The issues are terminal vacuum
value and pumping time. A scroll pump takes longer
to pump down and typically it ends at about 300mT.
Therefore, a turbo (MDP) is needed to obtain ultimate
vacuum with bleed of 15-20mT. A mechanical pump
typically will go down to 30mT with bleed (Ar).
And, it will do it much faster. Depending on the SEM,
you will see Au and Au/Pd. I don't see Pd or Ir or Pt.
The thickness is another variable. It doesn't take much
coating to eliminate charging. The tilt and rotate produces
a very even coating.

There are other brands of scroll pumps but I am only
familiar with the Edwards. It is easy to maintain
when it is necessary to replace the scroll lacing.
I think a rebuild kit is about $160.

Depending on your budget, I would certainly recommend the
Denton Desk V with turbo and scroll pump. Reliable, easy
to use and ultra fine coatings (get the tilt and rotate
option).

Not a seller--just a happy user. Any other question, pls
contact me.

gary g.


At 01:41 PM 2/10/2010, you wrote:



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From: nizets2-at-yahoo.com
Date: Thu, 11 Feb 2010 01:57:56 -0600
Subject: [Microscopy] Problem of TEM embedding of rat brain tissue

Contents Retrieved from Microscopy Listserver Archives
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Hi!
 
I can assure you that fixation has absolutely no effect on dehydration and embedding. There is no theoretical reason and that is confirmed in practice. I have been asked to embed very badly conserved material (one was a piece of rat brain!) and I was able to process the tissue as usual and to make 80-100 nm thin sections without problems.
 
Since, in theory, your protocol looks OK, I would rather troubleshoot every single possible practical problem, especially those which are not expected to happen. This really looks like a dehydration problem.
 
1)    Make sure than your resin is OK: ultracut empty blocks, without tissue (or embed something dry and soft, like a tiny piece of cloth). You can also simply try to cut the other end of already prepared blocks. If your resin is OK, then it is a problem of dehydration.
2)    Did the student work alone? Perhaps you should assist her and see if she’s doing everything all right. Perhaps during dehydration she leaves too much liquid and take it over to the next step.
3)    Ethanol contaminated? Try dehydration in acetone! This is more “extractive†than ethanol, but it also dehydrates very well and that’s the current problem you have to solve. You can do acetone/epon mixes, no need for a propylene oxide step.
4)    Troubleshoot the ultramicrotome: can you cut thin sections from a previously embedded material?
5)    Pure resin incubation: I usually do first 3 hours, then overnight, then another 3hours and then embedding. 2 changes for a total of 12 hours is perhaps a bit short.
6)    Extend the polymerization time: at least 2 days. In practice I just start on Friday and I have nice blocks on Monday.
 
Good luck

Stephane



----- Original Message ----
X-from: "ZZhang-at-uwyo.edu" {ZZhang-at-uwyo.edu}
To: nizets2-at-yahoo.com
Sent: Tue, February 9, 2010 8:10:54 PM

Hi All:

I have this 'weird' problem and need your help.

A student here has been trying do some TEM work with rat brain tissue. She first tried the "standard" protocol for embedding:

- Perfusion with 2% PFA, plus 2% GA
- Further fixation with 2% PFA, plus 2% GA, room temp, 1 hour
- 2nd fixation with 1% osmium, room temp, 1 hour
- Dehydration with EtoH (30....100%)
- Transition with propylene oxide
- Infiltration with 2:1, 1:2 (propylene oxide:resin), then 100% resin
- Polymerization (65C overnight)

The problem is that I always saw an incomplete infiltration (at least I thought so) - We were unable to cut even 2 um section. The sections fall apart, once it reaches the tissue. As I checked the semi-thin section, I saw holes all over the tissue.

I then asked the student tried to

1) extend the dehydration time (up to 3 min each step), and infiltration time (overnight each step)
2) additional microwave for infiltration;
3) smaller tissue (200 um vibritome section, instead of 1 mm cubes);
4) different resin (spur, embed812 and even LR white)

What is amazing to me is that NOTHING worked so far......

Any help?

Thank you.

Zhaojie Zhang, Ph.D.
Director, Microscopy Core Facility
Department of Zoology and Physiology
University of Wyoming
Laramie, WY 82071
zzhang-at-uwyo.edu
307-766-3038




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From: optik.a-at-a1.net
Date: Thu, 11 Feb 2010 15:46:33 -0600
Subject: [Microscopy] viaWWW: Ion beam milling gun lifetimes

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Email: optik.a-at-a1.net
Name: Andres Noah

Title-Subject: [Filtered] Ion beam milling

Message: Does anyone know the ion gun lifetime of:
Jeol Cross Section Polisher SM-09010
Hitachi Ion Milling System E-3500
Thank you
Andres


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From: weis183-at-yahoo.fr
Date: Fri, 12 Feb 2010 04:30:03 -0600
Subject: [Microscopy] Re : viaWWW: Ion beam milling gun lifetimes

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

Hi Andres,

I'm in charge of a JEOL CP.

We have bought the CP three years ago (roughly 400 samples ion polished) and we didn't change any part of the gun.
However the gun (and the chamber) has to be carrefully cleaned every 10-15 samples in order to get rid of the contamination. It takes usually one hour to clean it with fine polishing paper and ethanol.

One electrode of the gun is prone to contamination and can be seen as a consumable, but its life time is long enough if it is not damaged during the cleaning process.
In fact it is mainly the shield plate that protect the sample which is a concern because it has to be changed every 20-40 samples and is expensive.

Patrick Weisbecker
LCTS- Pessac
France

 
Title-Subject: [Filtered] Ion beam milling

Message: Does anyone know the ion gun lifetime of:
Jeol Cross Section Polisher SM-09010
Hitachi Ion Milling System E-3500
Thank you
Andres






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From: randy.burgess-at-hp.com
Date: Fri, 12 Feb 2010 12:58:07 -0600
Subject: [Microscopy] Ion Bean Milling

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Hello,
I was wondering if anyone could share similar information about the Leica EM TIC020.
Thank you,
Randy Burgess
HP Corvallis

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From: m_raven-at-lifesci.ucsb.edu
Date: Fri, 12 Feb 2010 14:15:27 -0600
Subject: [Microscopy] Cell fixation question

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This might not be the best group for the question. but, does anyone have
any suggestions on cell culture fixation protocols?

1) The importance of sucrose in the fixation solution
2) The percentage of paraformaldehyde

as it relates to the following?

I've seen some paraformaldehyde recipes that include sucrose and I
wonder if this is for simultaneous fixing AND sucrose protection of
tissues destined for freezing and processing on a cryostat? I've been
fixing a monolayer of stem cells grown on matrigel-coated tissue culture
treated polystyrene with 4% PAF (in 0.1M Cacodylate buffer, pH7.4), in a
very small surface area (~100mm^2, with a very small volume (50ul), and
am finding that the fix step is where I'm losing a lot of cells,
regardless of how gentle the pipetting goes. I'm thinking about trying
a PAF/sucrose mix to add density to the solution and to maybe provide
the cells with a hit of sugar as they're being fixed. Am also
considering dropping the PAF% to 2%

Thanks for any thoughts
Mary

--
Mary Raven
Integrated Microscopy Facility Director
Neuroscience Research Institute &
Molecular, Cellular & Developmental Biology
The University of California
Santa Barbara, CA 93106-5060

https://ucsb-microscopy.ucsd.edu/schedule/
http://www.lifesci.ucsb.edu/mcdb/research/facilities/microscopy/microscopy.html

Phone: (805) 893 8702
Fax: (805) 893 2005



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From: eldammar-at-hotmail.com
Date: Sat, 13 Feb 2010 16:42:57 -0600
Subject: [Microscopy] viaWWW: Hitachi SEM (S-4800)/ STEM

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Email: eldammar-at-hotmail.com
Name: El-Desouky Ammar

Organization: ARS, USDA

Title-Subject: [Filtered] STEM

Message: Hi everyone
We recently obtained a Hitachi SEM (S-4800) with an attachment to do
TEM (or STEM). Does anyone have experience using this machine for
TEM/STEM of biological samples (thin sections or negatively stained
preps)? How does it compare to the real TEM machines, can you get
good resultion at high mags (how high can you go), and under what
voltage (this machine goes only up to 30Kv), working distance and
other conditions? I'd appreciate hearing your opinions / experiences
on this matter. Thanks.



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From: g-jackson-at-ti.com
Date: Sat, 13 Feb 2010 16:43:27 -0600
Subject: [Microscopy] viaWWW: Epoxy bubbles

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Email: g-jackson-at-ti.com
Name: Guy Jackson

Organization: TI

Title-Subject: [Filtered] Epoxy bubbles

Message: A colleague of mine inquired about epoxy encapsulating a
MEMS device for cross sectioning. The device has interlaced fingers
approximately 20 um tall attached by cantilever springs. The fingers
are suspended in air above the device substrate. She has been
attempting to epoxy encapsulate the part for lapping but has been
plagued with bubbles being trapped between the fingers.

Can anyone suggest how to accomplish epoxy encapsulation while
minimizing the air bubbles?

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From: wesaia-at-iastate.edu
Date: Sat, 13 Feb 2010 17:56:34 -0600
Subject: [Microscopy] viaWWW: Epoxy bubbles

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Is she using vacuum to remove most of the air? We have a device from Struers for that purpose. It should reduce the size of the bubbles if not eliminate them.

There are also devices to apply pressure to the epoxy after pouring. That could be used after vacuum impregnation and would serve to reduce the size of the remaining bubbles even further.

I am not surprised that bubbles are trapped without these techniques. However, sometimes, a suitably non-viscous epoxy can be poured slowly enough to allow the air to move out of the way. Some epoxy formulations are quite viscous and it may not be possible to pour them slow enough to get rid of the bubbles.
________________________________________
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Email: g-jackson-at-ti.com
Name: Guy Jackson

Organization: TI

Title-Subject: [Filtered] Epoxy bubbles

Message: A colleague of mine inquired about epoxy encapsulating a
MEMS device for cross sectioning. The device has interlaced fingers
approximately 20 um tall attached by cantilever springs. The fingers
are suspended in air above the device substrate. She has been
attempting to epoxy encapsulate the part for lapping but has been
plagued with bubbles being trapped between the fingers.

Can anyone suggest how to accomplish epoxy encapsulation while
minimizing the air bubbles?

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From: gary-at-gaugler.com
Date: Sat, 13 Feb 2010 18:00:29 -0600
Subject: [Microscopy] Re: viaWWW: Epoxy bubbles

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Put the specimen in a vacuum chamber before the epoxy
sets up. The air will bubble out rather quickly. Best
not to use regular 5 minute epoxy. 15 minute stuff
works well.

A large chamber nor large capacity pump are necessary.
I use a plastic chamber from Duniway Vacuum and an
Edwards RV8 (overkill).

gary g.


At 02:45 PM 2/13/2010, you wrote:



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From: baskin-at-bio.umass.edu
Date: Sat, 13 Feb 2010 20:06:27 -0600
Subject: [Microscopy] Re: Hitachi SEM (S-4800)/ STEM

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

Hi,
The rule of thumb is that typical 60 - 80 nm ultra thin
sections are too thick for STEM on a FESEM, although there might be
exceptions. But samples spread on thin films that might otherwise be
negative stained can be looked at to great effect with STEM in your
instrument.

Have you asked your Hitatchi rep? They should be able to give
you a fairly infomative demo, at least to get you started.

Good luck,
Tobias

}
}
} Email: eldammar-at-hotmail.com
} Name: El-Desouky Ammar
}
} Organization: ARS, USDA
}
} Title-Subject: [Filtered] STEM
}
} Message: Hi everyone
} We recently obtained a Hitachi SEM (S-4800) with an attachment to do
} TEM (or STEM). Does anyone have experience using this machine for
} TEM/STEM of biological samples (thin sections or negatively stained
} preps)? How does it compare to the real TEM machines, can you get
} good resultion at high mags (how high can you go), and under what
} voltage (this machine goes only up to 30Kv), working distance and
} other conditions? I'd appreciate hearing your opinions / experiences
} on this matter. Thanks.
}


--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Biology Department
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/ / / \ \ \ University of Massachusetts
/ / / \ \ \ Amherst, MA, 01003
/ / ___ / \ \__/ \ ____
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Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

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From: dac-at-research.umass.edu
Date: Sun, 14 Feb 2010 08:04:57 -0600
Subject: [Microscopy] Re: viaWWW: Epoxy bubbles

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

Hi Guy,

If the materials are compatible with a solvent like actone (etc.) you
can use a dilution of the epoxy with solvent to aid wetting and
penetration and then let the solvent evaporate in a fume hood draft to
thicken the resin. We use this method for infiltrating fixed biological
samples but it should help in this case as well. After the resin has
reached ~ 100%, remove as much of the resin as possible and replace with
fresh resin and allow some time (preferably on a rotator - ~ 10rpm - to
mix. A vacuum can be applied at the end as others have described.

If you have a choice, use one of the low viscosity resins like the Ellis
modification of the original Spurr's formulation, and even the Low
Viscosity variant using Quetol 651, if needed.


Dale

} Spurr-replacement "New Formulation"
} E. Ann Ellis, Microscopy Today, Vol 14, No 4, July 2006
}
} Microscopy Today, July 2006, E. Ann Ellis "Solutions to the
} Problem of Substitution of ERL 4221 for Vinyl Cyclohexene Dioxide in
} Spurr Low Viscosity Embedding Formulations"
}
}
}
} All formulas are for a "10g batch" - with reference to (epoxy + acid anhydride)
}
} Epoxy Anhyride Catalyst Comments
} -------- -------- --------- -------- --------------
}
} ERL 4221 NSA DER-736 DMAE weights are in grams
}
} 4.10 5.90 0.95 0.10 Hard
} 4.10 5.90 1.43 0.10 Standard
} 4.10 5.90 1.90 0.10 Soft
}
}
} Low Viscosity Formula (Half the viscosity of the "Refomulated Spurr's")
} -------------------------
} ERL 4221 2.22g
} Quetol-651 1.40g
} NSA 6.38g
} DER-736 1.43g
} BDMA 0.2g
} Some think BDMA should be less, 0.1g
} I (DAC) used 0.09g DMAE (5 drops from a Samco #241 pipet) and it set nicely in 16h -at- 70C



g-jackson-at-ti.com wrote:
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} Email: g-jackson-at-ti.com
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} Title-Subject: [Filtered] Epoxy bubbles
}
} Message: A colleague of mine inquired about epoxy encapsulating a
} MEMS device for cross sectioning. The device has interlaced fingers
} approximately 20 um tall attached by cantilever springs. The fingers
} are suspended in air above the device substrate. She has been
} attempting to epoxy encapsulate the part for lapping but has been
} plagued with bubbles being trapped between the fingers.
}
} Can anyone suggest how to accomplish epoxy encapsulation while
} minimizing the air bubbles?
}
} Login Host: 192.94.94.106
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From: nizets2-at-yahoo.com
Date: Mon, 15 Feb 2010 04:05:37 -0600
Subject: [Microscopy] Cell fixation question

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

Hi Mary!

As the protocols to be used are very much depending on the goal pursed, it is very hard to make suggestions without knowing what you want to do.
Do you want to do cryo-embedding? light microscopy or electron microscopy?
I don't think that fixing is a small volume is a good idea. It is generally admitted that the volume of fixative should be 500x bigger than the volume of tissue fixed. Event though the volume of a cell monolayer is not very easy to estimate, I suppose that it won't be difficult to place your 1cm² specimen in a Petri dish and fix with 3-4 ml of fixative.
Otherwise I see no reason to lose cells during fixation. What is the temperature of the fixative? 37°C or 4°C?

Best regards,

Stephane



----- Original Message ----
X-from: "m_raven-at-lifesci.ucsb.edu" {m_raven-at-lifesci.ucsb.edu}
To: nizets2-at-yahoo.com
Sent: Fri, February 12, 2010 9:19:07 PM

This might not be the best group for the question. but, does anyone have
any suggestions on cell culture fixation protocols?

1) The importance of sucrose in the fixation solution
2) The percentage of paraformaldehyde

as it relates to the following?

I've seen some paraformaldehyde recipes that include sucrose and I
wonder if this is for simultaneous fixing AND sucrose protection of
tissues destined for freezing and processing on a cryostat?  I've been
fixing a monolayer of stem cells grown on matrigel-coated tissue culture
treated polystyrene with 4% PAF (in 0.1M Cacodylate buffer, pH7.4), in a
very small surface area (~100mm^2, with a very small volume (50ul), and
am finding that the fix step is where I'm losing a lot of cells,
regardless of how gentle the pipetting goes.  I'm thinking about trying
a PAF/sucrose mix to add density to the solution and to maybe provide
the cells with a hit of sugar as they're being fixed.  Am also
considering dropping the PAF% to 2%

Thanks for any thoughts
Mary

--
Mary Raven
Integrated Microscopy Facility Director
Neuroscience Research Institute &
Molecular, Cellular & Developmental Biology
The University of California
Santa Barbara, CA 93106-5060

https://ucsb-microscopy.ucsd.edu/schedule/
http://www.lifesci.ucsb.edu/mcdb/research/facilities/microscopy/microscopy.html

Phone: (805) 893 8702
Fax: (805) 893 2005



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From: John.Mardinly-at-wdc.com
Date: Mon, 15 Feb 2010 11:01:04 -0600
Subject: [Microscopy] viaWWW: Hitachi SEM (S-4800)/ STEM

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

When I worked at Intel, there were a number of factories that had no TEM but
relied on a Hitachi 5000 or 900 with a STEM detector for TEM mostly because
the managers were too damn cheap to buy a real TEM. 30KV for materials
science sounds just disgusting to us purists, but the crazy thing is that
those machines actually worked looking at FIB cross-sections of
microprocessors! Well, a 4800 is not a 5000, but should be pretty close to a
900, so don't count the thing out until you have tried it.

John Mardinly

Western Digital
-----Original Message-----
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Sent: Saturday, February 13, 2010 2:56 PM
To: John Mardinly

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Email: eldammar-at-hotmail.com
Name: El-Desouky Ammar

Organization: ARS, USDA

Title-Subject: [Filtered] STEM

Message: Hi everyone
We recently obtained a Hitachi SEM (S-4800) with an attachment to do
TEM (or STEM). Does anyone have experience using this machine for
TEM/STEM of biological samples (thin sections or negatively stained
preps)? How does it compare to the real TEM machines, can you get
good resultion at high mags (how high can you go), and under what
voltage (this machine goes only up to 30Kv), working distance and
other conditions? I'd appreciate hearing your opinions / experiences
on this matter. Thanks.



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From: danieleds-at-aol.com
Date: Mon, 15 Feb 2010 15:10:53 -0600
Subject: [Microscopy] viaWWW: Deben Stage control motor SEM

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Email: danieleds-at-aol.com
Name: Daniel Connors

Organization: EDS Services Inc

Title-Subject: [Filtered] Deben Stage control motor SEM

Message: Hi all:
I am looking for a used Deben Sprite stage controll motor. For
SEM stage controll. If any one has an old sysytem setting around
would like to here from you.
Thanks Dan connors

EDS Services Inc
1250 Corning Ave NW
Palm Bay, Fl. 32907

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From: murphyjudy-at-comcast.net
Date: Mon, 15 Feb 2010 18:44:56 -0600
Subject: [Microscopy] Betty Mathews 1927 - 2010

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

ELIZABETH "Betty" MATHEWS: 1927 - 2010

Elizabeth 'Betty' Mathews died January 26, 2010. She was a pioneering
faculty member at Delta College from 1970 to 1991 and was the first
biological instructor in the nationally known Electron Microscopy
program. Dr. Mathews received her Ph.D. from the University of
California, Berkeley in 1945. She worked in biological electron
microscopy for the next 46 years, publishing, teaching and nurturing
the students and program at Delta.

She is survived by her husband of 32 years, Don Mathews, also a Delta
retired faculty member.

Don and Betty retired to Carmel, California after their careers at
Delta to enjoy the ocean, golfing, and relaxation. A memorial service
to honor Betty will be held Saturday, February 20, 2010 at the Blessed
Sacrament Chapel, Carmel Mission, Carmel at 2 pm. A reception will be
held in Crespi Hall following the Memorial.

The family has requested that no flowers or gifts be sent. The family
suggests that donations in Betty's memory be sent to the Alzheimer's
Association of Monterey County, the Shriner's Hospital, or the
American Heart Association.

Betty was forever the optimist, an attitude that so many of her students carried forward.
She believed in her students and empowered them to do great things with their lives.
She contributed to microscopy education as well as service to the Electron Microscopy
Society of America in various functions over the years.

She will be dearly missed.


Judy Murphy
Stockton, CA


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From: john.robson-at-boehringer-ingelheim.com
Date: Tue, 16 Feb 2010 08:01:44 -0600
Subject: [Microscopy] RE: viaWWW: Epoxy bubbles

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

Hello Guy,

After reading some of the responses to your dilemma I would also like to add
one suggestion. You might want to place the epoxy in a vacuum chamber prior
to embedding your samples. You would be surprised how much out gassing you
will encounter. FYI in my experience the longer and higher the vacuum, at
his point, the better. BE forewarned, use a container that is significantly
large, the frothing epoxy has been known to overflow the brim! By
pre-evacuating the resin you will also find that your samples are less likely
to be disturbed by the movement of air bubbles once they are embedded and
re-introduced to the vacuum.

I would also consider brief sonication but this avenue is totally sample
dependent and might not be well suited to your applications?

Regards,

John Robson
Boehringer Ingelheim Pharmaceuticals, Inc.
900 Ridgebury Rd. / PO Box 368
Ridgefield, CT 06877
Phone (203)798-5640


==============================Original Headers==============================
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From: dac-at-research.umass.edu
Date: Tue, 16 Feb 2010 08:33:22 -0600
Subject: [Microscopy] Re: : Epoxy bubbles, vacuum and epoxy

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

Slight change in the subject line....

This topic has brought to mind a concern that evacuation may change the
composition of the resin. I have never done an experiment on this, but
since we can smell the components of the resin, the catalyst and also
the other components, especially with the low viscosity mixes, I have
been concerned that we are changing the composition of the mix when we
evacuate. Is it possible that the bubbles are not just air, but maybe a
resin component? Is there any evidence that this can be overdone
resulting in a change in the resin characteristics?

Dale

Dale Callaham
Umass -at- Amherst



john.robson-at-boehringer-ingelheim.com wrote:
} ----------------------------------------------------------------------------
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} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hello Guy,
}
} After reading some of the responses to your dilemma I would also like to add
} one suggestion. You might want to place the epoxy in a vacuum chamber prior
} to embedding your samples. You would be surprised how much out gassing you
} will encounter. FYI in my experience the longer and higher the vacuum, at
} his point, the better. BE forewarned, use a container that is significantly
} large, the frothing epoxy has been known to overflow the brim! By
} pre-evacuating the resin you will also find that your samples are less likely
} to be disturbed by the movement of air bubbles once they are embedded and
} re-introduced to the vacuum.
}
} I would also consider brief sonication but this avenue is totally sample
} dependent and might not be well suited to your applications?
}
} Regards,
}
} John Robson
} Boehringer Ingelheim Pharmaceuticals, Inc.
} 900 Ridgebury Rd. / PO Box 368
} Ridgefield, CT 06877
} Phone (203)798-5640
}
}
} ==============================Original Headers==============================
} 6, 29 -- From prvs=6561011c0=john.robson-at-boehringer-ingelheim.com Tue Feb 16 08:01:44 2010
} 6, 29 -- Received: from mail2.boehringer-ingelheim.com (mail2.boehringer-ingelheim.com [148.188.9.172])
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} 6, 29 -- Date: Tue, 16 Feb 2010 09:01:25 -0500
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From: john.robson-at-boehringer-ingelheim.com
Date: Tue, 16 Feb 2010 08:52:12 -0600
Subject: [Microscopy] Re: : Epoxy bubbles, vacuum and epoxy

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

Hello Dale,

I suppose it's possible but in my experience I have never encountered
problems after evacuating the resin. Of course we have been careful not to
extend the evacuation period too long to avoid overlap with the initiation of
curing. If anyone has encountered problems following resin evacuation I
would be grateful if they shared their experiences.

Thank You,

John Robson
Boehringer Ingelheim Pharmaceuticals, Inc.
900 Ridgebury Rd. / PO Box 368
Ridgefield, CT 06877
Phone (203)798-5640



-----Original Message-----
X-from: dac-at-research.umass.edu [mailto:dac-at-research.umass.edu]
Sent: Tuesday, February 16, 2010 9:40 AM
To: Robson,John AN BIP-US-R

Slight change in the subject line....

This topic has brought to mind a concern that evacuation may change the
composition of the resin. I have never done an experiment on this, but
since we can smell the components of the resin, the catalyst and also
the other components, especially with the low viscosity mixes, I have
been concerned that we are changing the composition of the mix when we
evacuate. Is it possible that the bubbles are not just air, but maybe a
resin component? Is there any evidence that this can be overdone
resulting in a change in the resin characteristics?

Dale

Dale Callaham
Umass -at- Amherst



john.robson-at-boehringer-ingelheim.com wrote:
}
----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
}
----------------------------------------------------------------------------
}
} Hello Guy,
}
} After reading some of the responses to your dilemma I would also like to
add
} one suggestion. You might want to place the epoxy in a vacuum chamber
prior
} to embedding your samples. You would be surprised how much out gassing you
} will encounter. FYI in my experience the longer and higher the vacuum, at
} his point, the better. BE forewarned, use a container that is significantly
} large, the frothing epoxy has been known to overflow the brim! By
} pre-evacuating the resin you will also find that your samples are less
likely
} to be disturbed by the movement of air bubbles once they are embedded and
} re-introduced to the vacuum.
}
} I would also consider brief sonication but this avenue is totally sample
} dependent and might not be well suited to your applications?
}
} Regards,
}
} John Robson
} Boehringer Ingelheim Pharmaceuticals, Inc.
} 900 Ridgebury Rd. / PO Box 368
} Ridgefield, CT 06877
} Phone (203)798-5640
}
}
} ==============================Original
Headers==============================
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16 08:01:44 2010
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From: DusevichV-at-umkc.edu
Date: Tue, 16 Feb 2010 09:19:20 -0600
Subject: [Microscopy] : Epoxy bubbles, vacuum and epoxy

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

Most of my specimens are tough to embed (demineralyzed or
non-demineralyzed hard tissues), so I routinely keep specimens for 1 hr
under vacuum after each change of the resin (epon substitute or spurr).
No ill effects observed.
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

} -----Original Message-----
} From: dac-at-research.umass.edu [mailto:dac-at-research.umass.edu]
} Sent: Tuesday, February 16, 2010 8:34 AM
} To: Dusevich, Vladimir
} Subject: [Microscopy] Re: : Epoxy bubbles, vacuum and epoxy
}
}
}
}
}
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} America
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}
} Slight change in the subject line....
}
} This topic has brought to mind a concern that evacuation may change
the
} composition of the resin. I have never done an experiment on this, but
} since we can smell the components of the resin, the catalyst and also
} the other components, especially with the low viscosity mixes, I have
} been concerned that we are changing the composition of the mix when we
} evacuate. Is it possible that the bubbles are not just air, but maybe
} a
} resin component? Is there any evidence that this can be overdone
} resulting in a change in the resin characteristics?
}
} Dale
}
} Dale Callaham
} Umass -at- Amherst
}
}
}
} john.robson-at-boehringer-ingelheim.com wrote:
} }
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} -------
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} }
} } Hello Guy,
} }
} } After reading some of the responses to your dilemma I would also
like
} to add
} } one suggestion. You might want to place the epoxy in a vacuum
} chamber prior
} } to embedding your samples. You would be surprised how much out
} gassing you
} } will encounter. FYI in my experience the longer and higher the
} vacuum, at
} } his point, the better. BE forewarned, use a container that is
} significantly
} } large, the frothing epoxy has been known to overflow the brim! By
} } pre-evacuating the resin you will also find that your samples are
} less likely
} } to be disturbed by the movement of air bubbles once they are
embedded
} and
} } re-introduced to the vacuum.
} }
} } I would also consider brief sonication but this avenue is totally
} sample
} } dependent and might not be well suited to your applications?
} }
} } Regards,
} }
} } John Robson
} } Boehringer Ingelheim Pharmaceuticals, Inc.
} } 900 Ridgebury Rd. / PO Box 368
} } Ridgefield, CT 06877
} } Phone (203)798-5640
} }
} }
} } ==============================Original
} Headers==============================
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} Feb 16 08:01:44 2010
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5, 26 -- From DusevichV-at-umkc.edu Tue Feb 16 09:19:19 2010
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5, 26 -- From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu}
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From: DusevichV-at-umkc.edu
Date: Tue, 16 Feb 2010 10:36:30 -0600
Subject: [Microscopy] : Epoxy bubbles, vacuum and epoxy

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

That sounds like it is the resin only. Perhaps the resin by itself has a lower vapor pressure. What level of vacuum are you applying?

I use Struers' SpeciFix-20 for mounting specimens. The mixed resin bubbles wildly below 100 mbar (if I recall correctly). I just tried pumping down some resin alone. It started bubbling around 50 mbar, but even at 20 mbar the bubbling was far from wild. There was no danger of overflowing the container. Perhaps it would have finished outgassing and settled down even more.

On the materials side, we work with the mixed resin from the beginning. It takes two to eight hours for the resin-hardener mix to setup at room temperature. I seem to recall some mixture starting to setup after just a few minutes under vacuum after much bubbling, so I assume something was changing with the composition. I just have not run any experiments to pin things down.

Warren S.

-----Original Message-----
X-from: DusevichV-at-umkc.edu [mailto:DusevichV-at-umkc.edu]
Sent: Tuesday, February 16, 2010 9:20 AM
To: wesaia-at-iastate.edu

I use vacuum oven, so pressure should be less than 100 mbar, but not by
much.

Vladimir


} -----Original Message-----
} From: wesaia-at-iastate.edu [mailto:wesaia-at-iastate.edu]
} Sent: Tuesday, February 16, 2010 9:35 AM
} To: Dusevich, Vladimir
} Subject: [Microscopy] RE: : Epoxy bubbles, vacuum and epoxy
}
}
}
}
}
-----------------------------------------------------------------------
} -----
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
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}
} That sounds like it is the resin only. Perhaps the resin by itself has
} a lower vapor pressure. What level of vacuum are you applying?
}
} I use Struers' SpeciFix-20 for mounting specimens. The mixed resin
} bubbles wildly below 100 mbar (if I recall correctly). I just tried
} pumping down some resin alone. It started bubbling around 50 mbar, but
} even at 20 mbar the bubbling was far from wild. There was no danger of
} overflowing the container. Perhaps it would have finished outgassing
} and settled down even more.
}
} On the materials side, we work with the mixed resin from the
beginning.
} It takes two to eight hours for the resin-hardener mix to setup at
room
} temperature. I seem to recall some mixture starting to setup after
just
} a few minutes under vacuum after much bubbling, so I assume something
} was changing with the composition. I just have not run any experiments
} to pin things down.
}
} Warren S.
}
} -----Original Message-----
} X-from: DusevichV-at-umkc.edu [mailto:DusevichV-at-umkc.edu]
} Sent: Tuesday, February 16, 2010 9:20 AM
} To: wesaia-at-iastate.edu
} Subject: [Microscopy] : Epoxy bubbles, vacuum and epoxy
}
} -------------------------------------
}
} Most of my specimens are tough to embed (demineralyzed or
} non-demineralyzed hard tissues), so I routinely keep specimens for 1
hr
} under vacuum after each change of the resin (epon substitute or
spurr).
} No ill effects observed.
} Vladimir
}
} Vladimir M. Dusevich, Ph.D.
} Electron Microscope Lab Manager
} 371 School of Dentistry
} 650 E. 25th Street
} Kansas City, MO 64108-2784
}
} } -----Original Message-----
} } From: dac-at-research.umass.edu [mailto:dac-at-research.umass.edu]
} } Sent: Tuesday, February 16, 2010 8:34 AM
} } To: Dusevich, Vladimir
} } Subject: [Microscopy] Re: : Epoxy bubbles, vacuum and epoxy
} }
} }
} }
} }
} }
}
-----------------------------------------------------------------------
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} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} } America
} } To Subscribe/Unsubscribe --
} } http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} }
}
-----------------------------------------------------------------------
} } -----
} }
} } Slight change in the subject line....
} }
} } This topic has brought to mind a concern that evacuation may change
} the
} } composition of the resin. I have never done an experiment on this,
} but
} } since we can smell the components of the resin, the catalyst and
also
} } the other components, especially with the low viscosity mixes, I
have
} } been concerned that we are changing the composition of the mix when
} we
} } evacuate. Is it possible that the bubbles are not just air, but
} maybe
} } a
} } resin component? Is there any evidence that this can be overdone
} } resulting in a change in the resin characteristics?
} }
} } Dale
} }
} } Dale Callaham
} } Umass -at- Amherst
} }
} }
} }
} } john.robson-at-boehringer-ingelheim.com wrote:
} } }
} ---------------------------------------------------------------------
} } -------
} } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} } America
} } } To Subscribe/Unsubscribe --
} } http://www.microscopy.com/MicroscopyListserver
} } } On-Line Help
} http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } }
} ---------------------------------------------------------------------
} } -------
} } }
} } } Hello Guy,
} } }
} } } After reading some of the responses to your dilemma I would also
} like
} } to add
} } } one suggestion. You might want to place the epoxy in a vacuum
} } chamber prior
} } } to embedding your samples. You would be surprised how much out
} } gassing you
} } } will encounter. FYI in my experience the longer and higher the
} } vacuum, at
} } } his point, the better. BE forewarned, use a container that is
} } significantly
} } } large, the frothing epoxy has been known to overflow the brim! By
} } } pre-evacuating the resin you will also find that your samples are
} } less likely
} } } to be disturbed by the movement of air bubbles once they are
} embedded
} } and
} } } re-introduced to the vacuum.
} } }
} } } I would also consider brief sonication but this avenue is totally
} } sample
} } } dependent and might not be well suited to your applications?
} } }
} } } Regards,
} } }
} } } John Robson
} } } Boehringer Ingelheim Pharmaceuticals, Inc.
} } } 900 Ridgebury Rd. / PO Box 368
} } } Ridgefield, CT 06877
} } } Phone (203)798-5640
}
}
} ==============================Original
} Headers==============================
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From: beaurega-at-westol.com
Date: Tue, 16 Feb 2010 11:07:50 -0600
Subject: [Microscopy] : Epoxy bubbles, vacuum and epoxy

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

Hi Dale,

When one uses a EPON 812 clone formulation, DDSA and NMA are also used in the mixture. Below is part of an unpublished article on vacuum embedding and capillary forces during infiltration. This part is really a demonstration about bubble formation.

"Pour DDSA into a long culture tube and fill it about half full. Put the test tube in a beaker in a vacuum chamber and slowly pull a vacuum on it. You will notice the formation of small bubbles. These are entrained or dissolved air. Before they reach the surface and break, release the vacuum. Notice what happens to the air bubbles. They just shrink in place and disappear. Next, immediately pull a vacuum and notice that they reappear in the same place. Stop increasing the vacuum and let them rise to the surface. Break them by applying and releasing the vacuum or using a micro spatula. Next, apply even more vacuum and that will cause larger bubbles to form. These are not air bubbles but vaporizing DDSA or whatever else is volatile in the DDSA. After you only get the large bubbles, release the vacuum. Using a micro spatula, gently disturb the DDSA by pushing it into the DDSA slightly and try not to entrain any air. Remove any bubbles you can see. Pull a vacuum on!
the D
DSA again. Small bubbles will form again. The larger bubbles will follow these smaller bubbles. This will give you insight as to what really happens in a micro porous membrane or porous agglomerated powder during vacuum impregnation. It also shows that DDSA will foam under vacuum. The more vacuum you apply, the larger the foam volume becomes.
{snip}
Are you really expelling air when you pull a vacuum? Maybe but maybe not. The DDSA will eventually expand under vacuum and find the path of least resistance to the outer surface of your sample. That path might not displace the air inside your agglomerates or membranes."
{snip}

Dale, I doubt that this vacuum will change the composition but it would be easy to determine by pulling a vacuum overnight on your epoxy components and observing any loss in volume. I don't believe you would remove a significant amount of any viscous "epoxy" component. Does that slight theoretical variation really matter?

If you asked, I bet you could get more formulations for various epoxies than the number of people receiving posting from this list server. Everyone seems to have their own formulation, at least for Epon 812 clones. So slight changes in composition are probably not that critical. Some only measure the components by volume with marked plastic micro beakers and not by weight. I always weighed mine. So there are variations in compsistion depending on the microtomist and his experiences. Some mix Araldite 502 with Epon 182 clone resin, for example.

Also the Epon 812 clone anhydrides, DDSA and NMA, are always absorbing water. They are changing every time you open the bottle. After some length of time, they have to be discarded or replaced.

"Is it possible that the bubbles are not just air, but maybe a resin component?" You can bet on that with Epon 812 clone epoxy formulations.

Paul Beauregard
Senior Research Associate, emeritus

At 08:33 AM 2/16/10 -0600, you wrote:
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From: tina-at-pbrc.hawaii.edu
Date: Tue, 16 Feb 2010 13:48:07 -0600
Subject: [Microscopy] : Epoxy bubbles, vacuum and epoxy

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

I always disliked pulling a vacuum on epoxy resin, and many, many years
ago I discovered I could minimize bubbles in epoxy by first holding the
molds and labels in the 60C oven overnight, or at least for a few hours
before using. I rarely get bubbles in anything that has gone through my
usual fixation (for biological samples). For materials (I currently have
paint chips in resin in the oven), I find that pre-warming the samples
also helps and, if this doesn't do it, pulling a vacuum on them before
embedding. I passed along this piece of wisdom to the mechanical
engineering guys who were getting bubbles in their epoxies containing
nanoparticles (another bane of my ultramicrotomy existence), and once they
started pre-drying their molds, they quit getting bubbles.

Aloha,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************


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From: lamiller-at-illinois.com
Date: Tue, 16 Feb 2010 14:48:31 -0600
Subject: [Microscopy] viaWWW: Bubbles in epoxy -- Vacuum usage

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Email: lamiller-at-illinois.com
Name: Lou Ann Miller

Organization: U of Illinois

Title-Subject: [Filtered] Bubbles in epoxy -- Vacuum usage

Message: I've used vacuum with epxoy mixtures for years, and have only mostly
seen benefits from it. However if left overnight in a pure epoxy
mixture, it will start to harden faster if not carefully watched.






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From: slc6-at-lehigh.edu
Date: Tue, 16 Feb 2010 14:49:03 -0600
Subject: [Microscopy] viaWWW: Research Engineer Position Lehigh University

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Email: slc6-at-lehigh.edu
Name: Sharon Coe

Organization: Lehigh University

Title-Subject: [Filtered] Research Engineer Position Lehigh University

Message: RESEARCH ENGINEER IN ADVANCED
ANALYTICAL ELECTRON MICROSCOPY

Department of Materials Science and Engineering
Lehigh University, Bethlehem, PA


The Nanocharacterization Laboratory at Lehigh
University houses an impressive array of
electron-optical instruments including two
state-of-the-art aberration-corrected analytical
electron microscopes. It is also the home of the
worldñrenowned Lehigh Microscopy School. The
Research Engineer position entails managing all
aspects of TEM/STEM operations within this user
facility. Major responsibilities of the
successful candidate include: training users in
the operation of the laboratoryís TEM/STEM
microscopes and related specimen preparation
equipment, including a dual-beam FIB; performing
routine maintenance and overseeing essential
repairs of the instrumentation; managing routine
operation, including logs, book-keeping, billing,
maintenance records, and safety; making
recommendations and assisting in the purchase,
installation and commissioning of new equipment;
liaison with external users from other
universities, government laboratories, and local
industry; providing outreach and tours for the
general public; and participating in the annual
Lehigh Microscopy School
(http://www.lehigh.edu/microscopy). Further
details of the Nanocharacterization Laboratory
and position requirements can be found at
http://www.lehigh.edu/~inmicro/.

Applicants should submit a complete CV and the
names of three referees to Sharon Coe, Department
of Materials Science and Engineering, Lehigh
University, 5 East Packer Avenue, Bethlehem, PA
18015-3195, USA. We will start reviewing
applications after March 15, 2010, and the search
will continue until the position is filled.
Lehigh University is committed to recruiting and
retaining women and members of minority groups.


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From: bozzola-at-siu.edu
Date: Tue, 16 Feb 2010 16:18:41 -0600
Subject: [Microscopy] Re: : Epoxy bubbles, vacuum and epoxy

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

Audrey Glauert also recommended heating the epoxy resin for several
minutes in the 60 degree oven prior to casting the specimens. This
decreases viscosity, increases the wetting capability of the resin and
diminishes bubbles.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

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2, 17 -- Subject: Re: [Microscopy] : Epoxy bubbles, vacuum and epoxy
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From: bozzola-at-siu.edu
Date: Tue, 16 Feb 2010 16:36:43 -0600
Subject: [Microscopy] : Epoxy bubbles, vacuum and epoxy

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

Dale Callaham (in a post earlier today) makes a good point (about
vacuum possibly removing the more volatile components of some resin
mixtures). I know this occurs since I have seen the components in the
vacuum pump oil when we changed the oil on a vacuum oven we regularly
used to outgas resin mixtures. The catalysts (like DMP 30 or DMAE) are
removed more easily that other components of epoxy resins and may
affect the resin mixture in the top layer. However, I doubt that the
short amount of vacuum exposure would adversely affect the entire
batch.

On a safety note, one must be aware that some resin components
(especially older Spurr formulations) are irritating and potential
carcinogens, so be careful handling the oil. Hopefully, the pump is
not exhausting directly into the room!

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

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From: lamiller-at-illinois.edu
Date: February 16, 2010 4:25:21 PM CST
Subject: [Microscopy] Re: : Epoxy bubbles, vacuum and epoxy

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

Hi John!
And all!


I find microwaving the specimen vial, or small quantities of epoxy
( too big amt... + hardens or explodes) for 10-20 seconds also helps
and probably for the same reasons too.

I have tried really hot ovens, like 100-110C with silicon molds, and I
actually find that this CREATES bubbles in the whole block.

If someone is encasing something that is metallic, perhaps also
lowering the temperature could help?



Lou Ann


Begin forwarded message:

X-from: bozzola-at-siu.edu

Audrey Glauert also recommended heating the epoxy resin for several
minutes in the 60 degree oven prior to casting the specimens. This
decreases viscosity, increases the wetting capability of the resin and
diminishes bubbles.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

==============================End of -
Headers==============================

{ { { { { { { { {} } } } } } } } } } } } } }
Lou Ann Miller, Service Supervisor
Center for Microscopic Imaging
College of Veterinary Medicine
University of Illinois MC=002

Room 1204 VMBSBld
2001 S Lincoln Ave
Urbana, IL 61821

217-244-1567
http://treefrog.cvm.uiuc.edu

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From: sergei2-at-ornl.gov
Date: Wed, 17 Feb 2010 12:37:54 -0600
Subject: [Microscopy] PFM workshop series

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

Dear Colleagues

We would like bring to your attention the workshop series on the
Piezoresponse Force Microscopy and Nanoscale Electromechanics of Polar
Materials. PFM has emerged as a powerful tool to explore and to study
nanoscale phenomena in ferroelectric and multiferroic devices, as well
as biological and polar macromolecular systems. Recently, PFM was
extended to study coupling between polarization and electronic
transport, as well as local phenomena in energy storage and generation
materials and strongly-correlated oxides, making it directly relevant to
the information and energy sciences and technology. The ubiquity of
electromechanical couplings in nanoscale systems – from molecular
electromotors to polar solids – suggests tremendous potential for this
technique.

This year, the three PFM workshops are planned for 2010 as follows:

7^th Workshop will be held in Montreal, Canada, May 31st – June 4th, as
a part of the Summer School on Advanced AFM Techniques and International
Workshop on Piezoresponse and Conductive AFM. The contact persons are
Andreas Ruediger, (ruediger-at-emt.inrs.ca) and Alexei Gruverman
(agruverman2-at-unlnotes.unl.edu).

8^th Workshop will be held in Beijing, China, August 25th – 27^th . The
information on the workshop can be found at
http://www.ustb.edu.cn/materials/files/pfm/International_Workshop.html.
The contact person is Prof. Jiangyu Li (jjli-at-u.washington.edu and
workshoppfm-at-yahoo.cn {mailto:workshoppfm-at-yahoo.cn} ).

9^th Workshop will be held in the conjunction with the International
Symposium on Ferroic Domains and Micro- to Nanoscopic structures
(ISFD-10) in Prague, Czech Republic, Sept. 22nd-24th. Please visit
http://palata.fzu.cz/isfd10/index.php?item=pfm for workshop details. The
contact person are /Jiří/ Hlinka (hlinka-at-fzu.cz {mailto:hlinka-at-fzu.cz} )
and Andrei Kholkin (kholkin-at-ua.pt {mailto:kholkin-at-ua.pt} ).

All three workshops, conveniently located in America, Europe, and Asia,
offer a combination of tutorial lectures by the advanced practitioners
of PFM, topical lectures by leading scientists in the field, industrial
lectures from PFM manufacturers, and poster sessions for attendees. It
also includes extensive lab demos for participants to gain hand-on
experience in PFM on various commercial systems, for which the attendees
can bring their own samples for testing and examinations. The PFM
conference (Prague) will also feature a series of invited and
contributed talks by the attendees.

On behalf of the PFM workshop series organizers:
Sergei V. Kalinin
Andrei Kholkin
Jiangyu Li

--
Sergei V. Kalinin
co-Theme Leader for Functional Imaging on the Nanoscale
The Center for Nanophase Materials Sciences
and Materials Sciences and Technology Division
Oak Ridge National Laboratory
Oak Ridge, TN 37922

Section Editor, Nanotechnology

Adjunct Associate Professor,
Department of Materials Science and Engineering,
University of Tennessee, Knoxville

Phone: (865) 241-0236
http://imaging.ornl.gov


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From: j.janssen-at-nki.nl
Date: Wed, 17 Feb 2010 14:36:28 -0600
Subject: [Microscopy] viaWWW: how to get skin embedded in a flat way?

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Email: j.janssen-at-nki.nl
Name: Hans Janssen

Organization: the Netherlands Cancer Institute

Title-Subject: [Filtered] how to get skin embedded in a flat way?

Message: I want to make parallel sections of epidermis from plastic
embedded mouse skin samples from which the hairs are removed with
cream. The problem is that during the embedding procedure the pieces
of skin tend to curl up, or get very wavy, so it is only possible to
get good sections of small aereas. Is this due to the keratinised
cells? Is there a way to press them down in a flat manner?
My procedure is: small pieces of skin fixed in Karnovsky fixative,
dehydration by ethanol series, propyleenoxide, prop/plastic mixtures,
DDSA/NMA/embed812/DMP30 embedding in flat molds.

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From: sergei2-at-ornl.gov
Date: Wed, 17 Feb 2010 15:17:03 -0600
Subject: [Microscopy] Research Staff Position - SPM of Energy Materials

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

Dear colleagues

It is my pleasure to bring to your attention the Research Staff Member
position in the field of Scanning Probe Microscopy of energy storage and
conversion materials opened in the Center for Nanophase Materials
Sciences at Oak Ridge National Laboratory. Below please find the text of
advertisement and the application information.

Yours

Sergei V. Kalinin

CNMS Research Staff for Imaging Functionality

Summary:
The Imaging Functionality Group at the Center for Nanophase Materials
Sciences (CNMS) of Oak Ridge National Laboratory is seeking a candidate
to fill a Research Staff position in the field of scanning probe
microscopy. The position offers an opportunity to take advantage of
state-ofthe- art instrumentation and recently developed capabilities for
piezoresponse force microscopy and a family of band excitation scanning
probe microscopies, including new spectroscopic modes for studies of
electrochemical dynamics and energy transfer in energy storage and
related materials, including oxides and ionic conductors. The CNMS
(http://cnms.ornl.qov/) is a collaborative nanoscience user research
facility established by the Office of Science, U.S. Department of
Energy. The CNMS has a diverse spectrum of nanoscience research activities
including a nanofabrication facility; laboratory-based research on
complex oxides, macromolecular materials, catalysts, functional
nanomaterials, bio-inspired nanomaterials, and transport;
characterization by electron microscopy, scanning probes, and neutron
diffraction and scattering; and theory, modeling, and simulation.

Qualifications:
This position requires a Ph.D. in the physical sciences, with an
emphasis on fundamental research in materials structure and function at
the nanoscale, particularly including atomic force microscopy. The
successful applicant must demonstrate ability to establish and lead a
research program in the experimental application, development, and
quantitative interpretation of modern scanning probe techniques for
studies of novel energy-related materials and systems. The applicant
must have the ability to work both independently and as part of a team
and to interact effectively with a broad range of colleagues. The
candidate will be engaged in the nanoscience user program, and must be
able to interact effectively in user-driven research and external
scientific collaborations. Presentations to an international scientific
audience and publication of scientific results in peer-reviewed journals
as lead author are necessary. Excellent oral and written communication
skills in English are required. All degree requirements must be
completed before starting the appointment.

Questions should be addressed to Art Baddorf, baddorfap-at-ornl.gov.
Applications will be accepted through the ORNL Jobs website at
http://jobs.ornl.gov/. From the right side of the website, choose
“View/Apply to Positions.” In the resulting search page, choose the
position category “Science-Physical Sciences.” In the resulting list,
this position is titled “Research Staff Member - Imaging Functionality”
and is position NC50205571. Review of applicants will begin immediately
and continue until the position is filled.

--
Sergei V. Kalinin
co-Theme Leader for Functional Imaging on the Nanoscale
The Center for Nanophase Materials Sciences
and Materials Sciences and Technology Division
Oak Ridge National Laboratory
Oak Ridge, TN 37922

Section Editor, Nanotechnology

Adjunct Associate Professor,
Department of Materials Science and Engineering,
University of Tennessee, Knoxville

Phone: (865) 241-0236
http://imaging.ornl.gov


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From: kenconverse-at-qualityimages.biz
Date: Wed, 17 Feb 2010 18:11:40 -0600
Subject: [Microscopy] User's manuals and schematics

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

Hi all,
I have a customer who just acquired a Hitachi S-2700 SEM. As is so often
the case, very little beyond the hardware arrived.

Actually, the manual and schematics for the SEM made it to the site, but
they are missing the operating instructions and schematics for the X-Y motor
driven stage (which I believe is Hitachi), along with the user's manual and
schematics for the GW Type 43 Infrared Chamberscope. I think the
Chamberscope's main problem is that most of the IR LEDs aren't working.

If anyone has any of these items and would be willing to share, I would
appreciate it. If you don't have it in electronic form, I can do that for
you and return both the original documents and the files.

Thanks,

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz






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From: vanessa.duke-at-sydney.edu.au
Date: Wed, 17 Feb 2010 19:36:25 -0600
Subject: [Microscopy] viaWWW: Job Opening University of Sydney SEM position

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Email: vanessa.duke-at-sydney.edu.au
Name: Vanessa Duke

Organization: The University of Sydney

Title-Subject: [Filtered] Job Announcement - The
University of Sydney - Scanning Electron
Microscopist

Message: Hello from Sydney!

My name is Vanessa Duke; I'm a Candidate Sourcing
& Research Analyst within the recruitment
division at the University of Sydney. I am
currently partnering with the Australian Key
Centre for Microscopy and Microanalysis in an
effort to locate a suitable candidate for the
appointment of Scanning Electron Microscopist.

When doing some preliminary research I discovered
this listserv and thought it would be a great
place to ask for some assistance from
professionals already working in the field...

Given the focus of the position is specimen
preparation, microscopy, microanalysis,
image/data analysis and intepretation I wanted to
get in touch with members of this list to see if
you were able to point me in the right direction
and possibly provide some advice on the best
avenues through which to get a communication out
to individuals who may be interested in a new
challenge here at the University of Sydney.

I am really looking for some advice on potential
advertising mediums, such as journals, forums and
associations and potentially any other
listerv/mailing list which may be helpful in
spreading the word...

A little more about the position:

The Australian Key Centre for Microscopy &
Microanalysis (AKCMM) is the headquarters of the
Australian Microscopy and Microanalysis Research
Facility (AMMRF), a major collaboration between
government and universities. The Centre
recognises the substantial role that microscopy,
imaging and microanalysis are set to play in the
next decade, as biotechnology and nanoscience
have increasing impact on science and society.
The Centre provides leadership, innovation and
ingenuity in Australian science and engineering
research.

The AKCMM is seeking a Scanning Electron
Microscopist to provide instruction, training and
support to users of the Centreís analytical
scanning electron microscope facilities. The
successful applicant will have demonstrated
expertise in specimen preparation techniques and
scanning electron microscopy as well as SEM
imaging to a broad range of biological and
engineered materials in different modes.
Experience in running and working in a multi-user
laboratory is required.


The full advertisement and application portal can
be viewed by following this link:
http://tinyurl.com/Scanning-Electron-Microscopist

Thanks for your time and I'm really looking
forward to hearing from you all shortly.

Regards,
Vanessa


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From: W.Muss-at-salk.at
Date: Thu, 18 Feb 2010 06:00:24 -0600
Subject: [Microscopy] RE: Embedding: how to get skin embedded in a flat way?

Contents Retrieved from Microscopy Listserver Archives
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Dear Hans Janssen,

understanding your problem of orthogonal sectioning proper of skin specimens to demonstrate follicle morphology after stripping / removal of hair(s) by a depilation cream I would like to tell you that IMO / in my experience the curling of skin specimen slices starts in higher ethanols due to tension forces of the dermal connective tissue lying beneath the basal keratinocytes.
I've never seen strong curling with specimens wich did not contain most of the dermal connective tissue and fat or contained such in a minimal amount.

Not knowing wether you must have / prepare very thin slices (i. e. nearest to a presumptive hair follicle) to easily get the right semithin sections for displaying what you want to see in morphology or you also could get started with somehow thicker tissue sclices I would like to say the following:

Just suggesting to prepare slices of at least 1 mm thickness (which is possible to achieve with sharp razor blades and correct use of stabile forceps [anatomical tweezers]), edge-dimension 2-3 mm in length, 3-4 mm in "heigth"- fix properly.
My routine skin specs - mostly all are punch biopsies - are prefixed as a whole (all done in vials on a specimen probe rotator) for at least 10-15 min at room temperature in FA-GA-buffer mixture (this is the time from the operating theatre to the lab), then divided into two halves, fixed again for at least 5-10 min and then - finally - sliced about 1 mm thickness [epidermis-dermis]; left in prefixative for at least another 30-60 minutes, final trimming of specs: separation of dermal tissue approx. 1-1.5 mm beneath the EDJ, then immersed in at least 4-5% buffered GA for at least 3 hours, over night at 4 degr.C, next morning: washing, osmication - starting dehydration 50% EtOH 70,80,90,96(2x),100(3x); instead of propyleneoxide, which perhaps supports the curling you observe: Acetonitrile (AN) as intermedium (3 x), AN:resin 1:1, final resin (3x)- embedding.

But YES, despite that protocol, some specs will show curling (assuming this to be a consequence of some properties of the connective tissue and / or its original density), but the extent isn't quite severe, so always we will have semithins over the whole area of the spec which was prepared initially.

Hope this is of any help to solve your embedding problem,

best of luck

Wolfgang MUSS
SALZBURG, AUSTRIA


==============================================================================================
OR Dr. phil. Wolfgang Muss
Head of EM-Lab
Institute of Pathology, SALK-LKH
(Salzburger Landeskliniken gemeinnuetzige GesmbH, Landeskrankenhaus = Fed. State Gen. Hosp.)
Muellner Hauptstrasse 48
A-5020 SALZBURG AUSTRIA/EUROPE
Web (German): www.salk.at

*)registered in: www.researchgate.net as "Wolfgang MUSS"
(NB.: Registration (free of charges) necessary to find / see
personal profile as well as my publication-collection)
ResearchGATE is the leading professional network for scientists.
It's free of charge and designed to meet researchers' needs
More than 200,000 scientists have joined!

and/or/alternatively (same Lab, same address):

Paracelsus Medical Private University (PMU)
Univ.-Institute of Pathology
Electron Microscopy Lab
Muellner Hauptstrasse 48
A-5020 SALZBURG, Austria/Europe
Web(German): http://www.pmu.ac.at
Phone work: +43+662+4482+4720
Mobile phone work: +43+662+4482-57704
Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o W.Muss")
E-Mail work: W.Muss-at-SALK.at
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PLEASE VISIT THE UPDATED WEBSITE of SCUR at } http://www.scur.org {
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} -----Ursprüngliche Nachricht-----
} Von: j.janssen-at-nki.nl [mailto:j.janssen-at-nki.nl]
} Gesendet: Mittwoch, 17. Februar 2010 21:40
} An: Muß Wolfgang
} Betreff: [Microscopy] Embedding: how to get skin embedded in a flat way?
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} Email: j.janssen-at-nki.nl
} Name: Hans Janssen
} Organization: the Netherlands Cancer Institute
} Title-Subject: how to get skin embedded in a flat way?
} Message:
}
} I want to make parallel sections of epidermis from plastic embedded mouse skin samples from which the hairs are removed with cream. The problem is that during the embedding procedure the pieces of skin tend to curl up, or get very wavy, so it is only possible to get good sections of small aereas. Is this due to the keratinised
} cells? Is there a way to press them down in a flat manner?
} My procedure is:
small pieces of skin fixed in Karnovsky fixative, dehydration by ethanol series, propyleenoxide, prop/plastic mixtures, DDSA/NMA/embed812/DMP30 embedding in flat molds.
} Login Host: 194.171.7.39
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}
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From: j.janssen-at-nki.nl
Date: Thu, 18 Feb 2010 06:25:06 -0600
Subject: [Microscopy] viaWWW: thanks for the answers on flat embedding skin

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Name: Hans Janssen

Organization: the Netherlands Cancer Institute

Title-Subject: [Filtered] thanks for the answers on flat embedding skin

Message: I like to thank the people that answered my question on flat
embedding skin. You gave me lots of different solutions that range
from keeping the tissue straight while fixing by means of filter
paper, cork or glass or aclar slides, to keeping it straight during
epon curing by means of clamping it in between glass or aclar
accompanied by pressing. I think I now can get beautiful sections.

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From: henrik.kaker-at-guest.arnes.si
Date: Thu, 18 Feb 2010 14:12:14 -0600
Subject: [Microscopy] viaWWW: HV problem-Strange noise

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Email: henrik.kaker-at-guest.arnes.si
Name: Henrik Kaker

Organization: SEM-EDS laboratory

Title-Subject: [Filtered] HV problem-Strange noise

Message: Dear All,

Today we found in our SEM Jeol JSM 35CF a "rattling" noise in the
upper part of the anode chamber where is the white ceramic part.
Please see the image, http://www.kaker.com/test/5.jpg. Microscope
condition: 25 kV, HV ON, Filament OFF, vacuum is OK. I am especially
interested about the reason for this nosie.


Henrik Kaker
SEM-EDS Laboratory
http://www2.arnes.si/~sgszmera1/index.html

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From: sandra.gardner-at-xerox.com
Date: Thu, 18 Feb 2010 14:12:40 -0600
Subject: [Microscopy] viaWWW: Digital Tablet for Particle size statistics

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Email: sandra.gardner-at-xerox.com
Name: Sandra Gardner

Organization: Xerox Research Centre of Canada

Title-Subject: [Filtered] Digital Tablet for Particle size statistics

Message: Hello All!

I am interested in using a digital tablet to collect particle size
data(ave particle size, aspect ratio, etc) from TEM images (directly
on digital image or on printed image if necessary for tablet). The
particles are sub micron in size and tend to agglomerate which makes
image analyses extremely challenging (many overlapping particles). I
feel that if I could outline the particles using a digital pen /
digital tablet I could then collect some semi-quantitative data.

I'm hoping someone out there has used this technique and can give me
some guidance. Direct advice or suggested publication of techniques
would be greatly appreciated. Any other suggestions for collecting
particle size data from TEM images would also be welcome (FYI, I am
familiar with Image Pro Plus and use it routinely for
non-agglomerated particles).

Cheers,
Sandra.

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From: cmshelton-at-srggi.com
Date: Thu, 18 Feb 2010 14:13:18 -0600
Subject: [Microscopy] viaWWW: Shopping for new Sputtercoater

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Email: cmshelton-at-srggi.com
Name: cathy

Organization: SRGGI

Title-Subject: [Filtered] Shopping for new Sputtercoater

Message: I am in the market for a new sputtercoater to prepare
samples for SEM. I current have a Pella Pelco SC-7 and would like to
try another brand.
When I need to return it to Pella to be serviced, they do not provide
a loaner. Doing without the coater for one day is a hardship. Any
recommendations would be greatly appreciated.
The loaner will not be such an issue with the new coater, as I will
keep the old one as a standby.

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From: protrain-at-emcourses.com
Date: Thu, 18 Feb 2010 14:49:31 -0600
Subject: [Microscopy] viaWWW: HV problem-Strange noise

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Hi

I am very confused by the rattling noise, could it be a tinkling noise due
to high voltage discharge? A check would be to lower the kV and see if the
noise became less, if it does it's discharge!

What causes high voltage discharge?

1. poor vacuum in the gun, this does not often show up on the penning
gauge as it may be positioned a long way from the gun area.
2. a dirt gun chamber (if it has an oily ozone smell that's discharge)
which should be cleaned until all the smell has gone away. Clean the walls
and the ceramic as I believe it is glazed therefore it is to be treated like
a metal surface. Remember ammonia solution will dissolve tungsten?

Check it out - good luck


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com


-----Original Message-----
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Email: henrik.kaker-at-guest.arnes.si
Name: Henrik Kaker

Organization: SEM-EDS laboratory

Title-Subject: [Filtered] HV problem-Strange noise

Message: Dear All,

Today we found in our SEM Jeol JSM 35CF a "rattling" noise in the
upper part of the anode chamber where is the white ceramic part.
Please see the image, http://www.kaker.com/test/5.jpg. Microscope
condition: 25 kV, HV ON, Filament OFF, vacuum is OK. I am especially
interested about the reason for this nosie.


Henrik Kaker
SEM-EDS Laboratory
http://www2.arnes.si/~sgszmera1/index.html

Login Host: 193.189.163.74
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From: Elliott-at-arizona.edu
Date: Thu, 18 Feb 2010 15:06:19 -0600
Subject: [Microscopy] Re: viaWWW: Digital Tablet for Particle size statistics

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Hi Sandra

What you are suggesting is quite simple. The only problem is that
which is EASY for the human is difficult for a computer.

I use Wacom tablets for 'manual data segmentation'. An easy (not the
best, but very easy to learn - I use high school students or
undergrads to do the work) way to do this is to open your image in
photoshop (or some such program). Using your digital pen or mouse,
select the pencil tool and set the tool size to something easy to use,
but smaller than your particle. Create a new layer. Draw the
particles you want to analyze in the new layer. Save this document.
This is the record of what will be analyzed. Create a new document
with only the drawing layer. Open this in NIH image (ImageJ) and
"Analyze Particles" You can get all kinds of information out of this
analysis.

If you want more info, please contact me.
David


On Feb 18, 2010, at 1:17 PM, sandra.gardner-at-xerox.com wrote:

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} Title-Subject: [Filtered] Digital Tablet for Particle size statistics
}
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}
} I am interested in using a digital tablet to collect particle size
} data(ave particle size, aspect ratio, etc) from TEM images (directly
} on digital image or on printed image if necessary for tablet). The
} particles are sub micron in size and tend to agglomerate which makes
} image analyses extremely challenging (many overlapping particles). I
} feel that if I could outline the particles using a digital pen /
} digital tablet I could then collect some semi-quantitative data.
}
} I'm hoping someone out there has used this technique and can give me
} some guidance. Direct advice or suggested publication of techniques
} would be greatly appreciated. Any other suggestions for collecting
} particle size data from TEM images would also be welcome (FYI, I am
} familiar with Image Pro Plus and use it routinely for
} non-agglomerated particles).
}
} Cheers,
} Sandra.
}
} Login Host: 13.12.254.95
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From: rbeavers-at-mail.smu.edu
Date: Thu, 18 Feb 2010 16:51:43 -0600
Subject: [Microscopy] Carbon measurments

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Group,

Looking at a polyimide film that has been scanned with a laser and it produces small 10 micron cone shaped peaks that we believe contains a higher concentration of carbon at the peak vs the base. Challenge is to measure the carbon variation which with some rough SEM/EDS work seems to be around 2wt%.

Geometry issues cause me to question EDS as the best way to attempt this.

Looking for another possible technique to try.

Roy Beavers
Southern Methodist University
Department of Earth Sciences
P.O. Box 750395
Dallas, TX  75275
Voice: 214-768-2756
Fax: 214-768-2701
Email: rbeavers-at-smu.edu



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From: m_raven-at-lifesci.ucsb.edu
Date: Thu, 18 Feb 2010 17:44:01 -0600
Subject: [Microscopy] LM - Workshop Announcement

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Advanced Microscopy and Digital Imaging Workshop
April 26-29, 2010
Integrated Microscopy Facility
Neuroscience Research Institute & the Department of Molecular Cellular
Developmental Biology
University of California, Santa Barbara

Enrollment is limited and early registration is encouraged.

Registration is $925. Scholarships are available on the basis of need.

Registration Deadline is April 9th, 2010

The Integrated Microscopy Facility operated by the Neuroscience Research
Institute (NRI) and the Department of Molecular, Cellular, and
Developmental Biology, offers a workshop to provide intensive training
with light microscopy. This workshop includes three and a half days of
seminars and hands on instruction. Through the course, participants will
gain theoretical and practical experience with light microscopy.
Lectures will focus on microscopy principals and applications while
laboratories will focus on collecting quality images and data for
research. Fixed biological sample will be used in this course although
many topics will be relevant for non-biological samples and live cell
imaging. This seminar serves as an excellent foundation foundation for
our live cell imaging course to be offered Oct 25-29, 2010.

All attendees will use fully equipped research grade microscopes with
the aid of trained instructors. These microscopes enable all students to
practice such light microscopy techniques as fluorescence, phase
contrast, Nomarski differential interference contrast and brightfield
imaging. All students will be exposed to advanced imaging techniques
including confocal and two-photon imaging. With a maximum enrollment of
12 students, there will be many opportunities to work with all of the
microscopes and cameras. Attendees will leave the course comfortable
with both the principals and practice of light microscopy.

For more information or to register visit:
https://www.lifesci.ucsb.edu/mcdb/events/imaging_workshop/

If you have any questions please feel free to contact Mary Raven at
805-893-8702 or UCSBmicroscopy-at-lifesci.ucsb.edu

--
Mary Raven
Integrated Microscopy Facility Director
Neuroscience Research Institute &
Molecular, Cellular & Developmental Biology
The University of California
Santa Barbara, CA 93106-5060

https://ucsb-microscopy.ucsd.edu/schedule/
http://www.lifesci.ucsb.edu/mcdb/research/facilities/microscopy/microscopy.html

Phone: (805) 893 8702
Fax: (805) 893 2005



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From: Noel.Goldsmith-at-dsto.defence.gov.au
Date: Fri, 19 Feb 2010 00:21:27 -0600
Subject: [Microscopy] High Resolution Coaters [SEC=UNCLASSIFIED]

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Hi,
Having an FE SEM, we are looking to purchase a high(er) resolution sputter
coater.
As it is years since I looked at the market I thought I would ask the
members of this list for a list.
The things I would like to know.
1. Makes available.
2. Support in Australia available.
3. What works.
4. What does not work.
5 Any other information which might be useful, such as reliability etc.
If you wish to reply off-list please do.
Thank you
Noel

Noel Goldsmith
Air Vehicles Division
DSTO
506 Lorimer Street
Port Melbourne
3207 Victoria
AUSTRALIA
613 96267527
0428364003
noel.goldsmith-at-dsto.defence.gov.au
--



IMPORTANT: This email remains the property of the Australian Defence Organisation and is subject to the jurisdiction of section 70 of the CRIMES ACT 1914. If you have received this email in error, you are requested to contact the sender and delete the email.


==============================Original Headers==============================
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From: stephan.handschin-at-emez.ethz.ch
Date: Fri, 19 Feb 2010 04:34:29 -0600
Subject: [Microscopy] viaWWW: Glue for HM20 resin

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Email: stephan.handschin-at-emez.ethz.ch
Name: Stephan Handschin

Title-Subject: [Filtered] Glue for HM20 resin

Message: I am looking for an instant glue which is able to glue
reoriented samples embedded in HM20 on a HM20-stick. We had some
mighty bond instant glue from a philipinian company, but it's not
available for us any more. The normal instant glues we test are not
able to glue strongly enough.
Thank you a lot in advance.
Kind regards

Login Host: 129.132.116.94
---------------------------------------------------------------------------

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From: ehaller-at-health.usf.edu
Date: Fri, 19 Feb 2010 07:49:13 -0600
Subject: [Microscopy] viaWWW: Glue for HM20 resin

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

Stephan,

I've been buying and using Permabond 910 (I buy mine from Electron Microscopy Sciences) for years for both HM20 and Epon equivalent epoxies. There is a difference between this and Mighty Bond or Crazy Glue, both of which get brittle and allow your block to fall off the dummy blocks. The Permabond does not get brittle, and can be cut with a razor blade. It actually melts the embedding media and causes it to re-harden. The glue can be sectioned, although its properties under the electron beam are not as good as those of the epoxy or methacrylate itself. I keep mine in the refrigerator, and a bottle lasts for years. I've found that the oil from my skin acts as an excellent "primer" before I join pieces of plastic together. I saw the piece of sample block out, sand the piece smooth, sand the "dummy" block surface to roughen the face to make for better adherance, coat both surfaces with a little oil from the skin of my nose (I know this sounds gross, but it works unbelievably well!), apply one small drop of glue, put the two pieces of plastic together, and they set almost immediately. I allow the glue to harden for several hours, then I'm ready to section. After drying, I wash the block in mild detergent, then water (after trimming) to remove skin oil, and I'm ready to go. It's important to sand both surfaces smooth before gluing to make sure both surfaces are flat and parallel to each other so there are no voids between the surfaces that could cause compression in your sections. I've used this technique for over 30 years now. If you have any questions on the technique, feel free to write. This is how I handle all of my cell pellet work.


Ed Haller


Edward Haller, Lab Manager
Integrative Biology Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-cas.usf.edu

________________________________________
X-from: stephan.handschin-at-emez.ethz.ch [stephan.handschin-at-emez.ethz.ch]
Sent: Friday, February 19, 2010 5:48 AM
To: Haller, Edward

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Email: stephan.handschin-at-emez.ethz.ch
Name: Stephan Handschin

Title-Subject: [Filtered] Glue for HM20 resin

Message: I am looking for an instant glue which is able to glue
reoriented samples embedded in HM20 on a HM20-stick. We had some
mighty bond instant glue from a philipinian company, but it's not
available for us any more. The normal instant glues we test are not
able to glue strongly enough.
Thank you a lot in advance.
Kind regards

Login Host: 129.132.116.94
---------------------------------------------------------------------------

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15, 37 -- From: "Haller, Edward" {ehaller-at-health.usf.edu}
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From: James.Passmore-at-sealedair.com
Date: Fri, 19 Feb 2010 08:05:53 -0600
Subject: [Microscopy] viaWWW: Digital Tablet for Particle size statistics

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

This can be done quite easily using just ImageJ, eliminating the need for
a second program and the extra steps.

1. With image in ImageJ (I assume you have calibrated it) outline a
particle, and press 't' to add it to the ROI Manager.
2. Continue outlining and pressing 't' for each particle. Be sure "show
all" is checked so you can see what you've already selected.
3. In the ROI Manager window, the 'More' button will show a choice to
save the selection set if you need it for record-keeping. Not necessary
for analysis.
4. Also in the ROI Manager, you can select "Multi Measure" to measure
each selected area

Note that I didn't get into image calibration or selection of which
measurements you want to perform on the particles. I just wanted to show
it was easily done. I'm sure Image Pro will also have the capability, but

I rarely use it so I can't give instructions.

----------------------------------------------
Jim Passmore
Research Associate
Sealed Air Corporation
james.passmore-at-sealedair.com
864-433-2927 voice
864-433-2205 fax
----------------------------------------------


Elliott-at-arizona.edu wrote on 02/18/2010 04:07:32 PM:

}
}
}
}
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}
} Hi Sandra
}
} What you are suggesting is quite simple. The only problem is that
} which is EASY for the human is difficult for a computer.
}
} I use Wacom tablets for 'manual data segmentation'. An easy (not the
} best, but very easy to learn - I use high school students or
} undergrads to do the work) way to do this is to open your image in
} photoshop (or some such program). Using your digital pen or mouse,
} select the pencil tool and set the tool size to something easy to use,
} but smaller than your particle. Create a new layer. Draw the
} particles you want to analyze in the new layer. Save this document.
} This is the record of what will be analyzed. Create a new document
} with only the drawing layer. Open this in NIH image (ImageJ) and
} "Analyze Particles" You can get all kinds of information out of this
} analysis.
}
} If you want more info, please contact me.
} David
}
}
} On Feb 18, 2010, at 1:17 PM, sandra.gardner-at-xerox.com wrote:
}
} }
} }
} }
} }
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} } Email: sandra.gardner-at-xerox.com
} } Name: Sandra Gardner
} }
} } Organization: Xerox Research Centre of Canada
} }
} } Title-Subject: [Filtered] Digital Tablet for Particle size statistics
} }
} } Message: Hello All!
} }
} } I am interested in using a digital tablet to collect particle size
} } data(ave particle size, aspect ratio, etc) from TEM images (directly
} } on digital image or on printed image if necessary for tablet). The
} } particles are sub micron in size and tend to agglomerate which makes
} } image analyses extremely challenging (many overlapping particles). I
} } feel that if I could outline the particles using a digital pen /
} } digital tablet I could then collect some semi-quantitative data.
} }
} } I'm hoping someone out there has used this technique and can give me
} } some guidance. Direct advice or suggested publication of techniques
} } would be greatly appreciated. Any other suggestions for collecting
} } particle size data from TEM images would also be welcome (FYI, I am
} } familiar with Image Pro Plus and use it routinely for
} } non-agglomerated particles).
} }
} } Cheers,
} } Sandra.
} }
} }
}


==============================Original Headers==============================
9, 16 -- From James.Passmore-at-sealedair.com Fri Feb 19 08:05:52 2010
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From: dac-at-research.umass.edu
Date: Fri, 19 Feb 2010 08:19:21 -0600
Subject: [Microscopy] viaWWW: Glue for HM20 resin

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

Look at the products from Balsa USA. They have CA glues with different
thickness, accelerators, removers, etc.

https://www.balsausa.com/store/category.php?id_category=33

Cheers,

Dale

D. Callaham
Umass -at- Amherst



ehaller-at-health.usf.edu wrote:
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} Stephan,
}
} I've been buying and using Permabond 910 (I buy mine from Electron Microscopy Sciences) for years for both HM20 and Epon equivalent epoxies. There is a difference between this and Mighty Bond or Crazy Glue, both of which get brittle and allow your block to fall off the dummy blocks. The Permabond does not get brittle, and can be cut with a razor blade. It actually melts the embedding media and causes it to re-harden. The glue can be sectioned, although its properties under the electron beam are not as good as those of the epoxy or methacrylate itself. I keep mine in the refrigerator, and a bottle lasts for years. I've found that the oil from my skin acts as an excellent "primer" before I join pieces of plastic together. I saw the piece of sample block out, sand the piece smooth, sand the "dummy" block surface to roughen the face to make for better adherance, coat both surfaces with a little oil from the skin of my nose (I know this sounds gross, but it works unbeliev
abl!
} y well!), apply one small drop of glue, put the two pieces of plastic together, and they set almost immediately. I allow the glue to harden for several hours, then I'm ready to section. After drying, I wash the block in mild detergent, then water (after trimming) to remove skin oil, and I'm ready to go. It's important to sand both surfaces smooth before gluing to make sure both surfaces are flat and parallel to each other so there are no voids between the surfaces that could cause compression in your sections. I've used this technique for over 30 years now. If you have any questions on the technique, feel free to write. This is how I handle all of my cell pellet work.
}
}
} Ed Haller
}
}
} Edward Haller, Lab Manager
} Integrative Biology Electron Microscopy Core
} SCA 110
} 4202 East Fowler Avenue
} Tampa, FL 33620
} (813)974-2676
} ehaller-at-cas.usf.edu
}
} ________________________________________
} X-from: stephan.handschin-at-emez.ethz.ch [stephan.handschin-at-emez.ethz.ch]
} Sent: Friday, February 19, 2010 5:48 AM
} To: Haller, Edward
} Subject: [Microscopy] viaWWW: Glue for HM20 resin
}
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} Email: stephan.handschin-at-emez.ethz.ch
} Name: Stephan Handschin
}
} Title-Subject: [Filtered] Glue for HM20 resin
}
} Message: I am looking for an instant glue which is able to glue
} reoriented samples embedded in HM20 on a HM20-stick. We had some
} mighty bond instant glue from a philipinian company, but it's not
} available for us any more. The normal instant glues we test are not
} able to glue strongly enough.
} Thank you a lot in advance.
} Kind regards
}
} Login Host: 129.132.116.94
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==============================Original Headers==============================
8, 22 -- From dac-at-research.umass.edu Fri Feb 19 08:19:21 2010
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From: marie.cantino-at-uconn.edu
Date: Fri, 19 Feb 2010 12:38:18 -0600
Subject: [Microscopy] SEM: flattening tissue samples for CPD

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

I was reminded by a recent posting (flattening samples for embedding)
of a problem we have recently encountered.

A student is trying to do SEM on the distal ends of critical point
dried hummingbird tongues. The main body of the tongue is about a mm
wide by 1-2 cm long, and bifurcates towards the end into two flat
ribbons of tissue that form spirals during critical point drying, even
if they are kept flat during the fixation and dehydration. We would
like to force them to stay flat all the way through the critical point
drying. Any sheets of material used to sandwich them would have to be
rigid enough to prevent curling (possibly by mounting in a frame) but
not mechanically damage the surface, and it would need to be stable
chemically and allow adequate exchange during critical point drying.
I would be interested in any experiences you have with different
materials for creating such a sandwich (e.g. metal or polymer meshes,
filter papers). I am also open to other ideas, but would prefer not
to pin them, as it is likely to damage critical parts.

Our protocol involves aldehyde and osmium fixation, followed by
dehydration to 100% ethanol and critical point drying. No other
solvents (besides the CO2) are used.

Marie

Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369


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From: baskin-at-bio.umass.edu
Date: Fri, 19 Feb 2010 14:28:05 -0600
Subject: [Microscopy] Re: SEM: flattening tissue samples for CPD

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

Marie,
I have had some success preventing though not minimizing
curling of plant tissues by encasing them between formvar films. I
make a wire loop, plunge it over a formvar rectangle, let dry, put my
sample flat on the film (when the sample is still hydrated) and then
plunge again over a second rectangle. Then the loop + formvar
sandwiched sample can be dehydrated and cpd no problem. When it comes
time to mount on a stub you have to be a little dextrous and tease
apart the formvar films but this is not so difficult with practice.
Everything gets through formvar and the only thing I have found to
dissolve it is propylene oxide.

Hope this helps,
Tobias

}
}
} I was reminded by a recent posting (flattening samples for embedding)
} of a problem we have recently encountered.
}
} A student is trying to do SEM on the distal ends of critical point
} dried hummingbird tongues. The main body of the tongue is about a mm
} wide by 1-2 cm long, and bifurcates towards the end into two flat
} ribbons of tissue that form spirals during critical point drying, even
} if they are kept flat during the fixation and dehydration. We would
} like to force them to stay flat all the way through the critical point
} drying. Any sheets of material used to sandwich them would have to be
} rigid enough to prevent curling (possibly by mounting in a frame) but
} not mechanically damage the surface, and it would need to be stable
} chemically and allow adequate exchange during critical point drying.
} I would be interested in any experiences you have with different
} materials for creating such a sandwich (e.g. metal or polymer meshes,
} filter papers). I am also open to other ideas, but would prefer not
} to pin them, as it is likely to damage critical parts.
}
} Our protocol involves aldehyde and osmium fixation, followed by
} dehydration to 100% ethanol and critical point drying. No other
} solvents (besides the CO2) are used.
}
} Marie
}
} Dr. Marie E. Cantino
} Associate Professor of Physiology and Neurobiology
} Director, Electron Microscopy Laboratory
} University of Connecticut, Unit 3242
} Phone: 860-486-3588
} Fax: 860-486-6369


==============================Original Headers==============================
4, 21 -- From baskin-at-bio.umass.edu Fri Feb 19 14:28:05 2010
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From: kim.e.graebner-at-boeing.com
Date: Fri, 19 Feb 2010 15:22:51 -0600
Subject: [Microscopy] viaWWW: Need attachment manual for SRT2 Scan / Tilt / Rotation

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Name: Kim Graebner

Title-Subject: [Filtered] Need attachment manual for SRT2 Scan / Tilt
/ Rotation Module

Message: I need an operation manual for a JEOL 35CF SEM attachment,
SRT2 Scan / Rotate / Tilt module. I need to see operation and more
importantly connection requirements.

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From: swatkins-at-pitt.edu
Date: Fri, 19 Feb 2010 15:23:21 -0600
Subject: [Microscopy] viaWWW: freeze fracture

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Email: swatkins-at-pitt.edu
Name: simon watkins

Organization: university of pittsburgh

Title-Subject: [Filtered] freeze fracture

Message: Hi all. We are attempting to buy a replacement/new freeze
fracture machine, but there dont seem to be any obvious suppliers
(cressington/leica/etc)
anyone out there got ideas, or better a compact system to sell us?
S

Simon C. Watkins Ph.D, FRC Path
Professor and Vice Chair Cell Biology and Physiology
Professor Immunology Director Center for Biologic Imaging
BSTS 225
University of Pittsburgh
3500 Terrace St
Pittsburgh PA 15261
412-352-2277
www.cbi.pitt.edu


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From: ralkhat-at-ncsu.edu
Date: Fri, 19 Feb 2010 15:23:40 -0600
Subject: [Microscopy] viaWWW: Automated Sample Plunging

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Email: ralkhat-at-ncsu.edu
Name: Rami Alkhatib

Organization: North Carolina State University

Title-Subject: [Filtered] Automated Sample Plunging

Message: Hello,
I am planning to buy an "Automated Sample plunging" for Freeze
fracture purposes. I am looking for an affordable system but good at
the same time. Any advice about which one should I get and where I
can get it.

Many thanks in advance!

Dr. Rami Alkhatib
North Carolina State University
Crop Science Department
Raleigh, NC 27695


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From: raristau-at-ims.uconn.edu
Date: Thu, 18 Feb 2010 14:19:28 -0600
Subject: [Microscopy] viaWWW: Digital Tablet for Particle size statistics

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Sandra,

Use a Sharpie pen on clear acetate sheet (remember the old overhead
projector sheets?) to trace the outline of your particles. Then digitally
scan the tracing. Not any slower than using a digital tablet to trace, and I
like it because I can move the acetate sheet after tracing one particle and
then trace the overlying particle fully onto a blank area of the sheet--no
overlapping particles on the tracing.

I am learning to write left-handed now because I have worn out my right hand
doing so many grain boundary tracings.

Roger

Roger A. Ristau, PhD
Electron Microscopy Specialist
Institute of Materials Science
97 North Eagleville Road
University of Connecticut
Storrs, CT 06269
vox: 860-486-5453
fax: 860-486-4745

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Email: sandra.gardner-at-xerox.com
Name: Sandra Gardner

Organization: Xerox Research Centre of Canada

Title-Subject: [Filtered] Digital Tablet for Particle size statistics

Message: Hello All!

I am interested in using a digital tablet to collect particle size
data(ave particle size, aspect ratio, etc) from TEM images (directly
on digital image or on printed image if necessary for tablet). The
particles are sub micron in size and tend to agglomerate which makes
image analyses extremely challenging (many overlapping particles). I
feel that if I could outline the particles using a digital pen /
digital tablet I could then collect some semi-quantitative data.

I'm hoping someone out there has used this technique and can give me
some guidance. Direct advice or suggested publication of techniques
would be greatly appreciated. Any other suggestions for collecting
particle size data from TEM images would also be welcome (FYI, I am
familiar with Image Pro Plus and use it routinely for
non-agglomerated particles).

Cheers,
Sandra.

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From: eikonika-at-otenet.gr
Date: Sat, 20 Feb 2010 03:09:20 -0600
Subject: [Microscopy] SEM: flattening tissue samples for CPD

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Dear Marie

This problem is really hardŠ During and after CPD even the slightest forces
applied to the biological sample will destroy the fine surface details. When
I CPD epithelial tissue, very often I find areas with marks created by the
basket mesh when the sample hits against it during flushing with CO2. So I
think that any attempt to sandwich your sample ­the bird tongue- will cause
extensive damage on it.

What can you do then.. I can only offer you ideas:

1. Cut with fine scissors the tongue in several cuts perpendicular to its
long axis. The cuts should be partial, leaving one end still attached
(looking like a single strand DNA in textbooks) so you will not lose the
anatomy and orientation. Then after CPD -do it immediately, as the sample is
still a bit elastic and not completely rigid- using fine forceps, very
carefully, align the sample. Avoid touching the surface but touch the cut
ends so the damage from forceps will not affect the surface.

2. Another idea (may be practically impossible but will be great if it
works!). Try to insert a pin (can be fine needle or some plant spike or
similar) starting from the basis of the tongue and follow the longitudinal
axis towards the end and try to enter one bifurcation as far as possible.
The idea is that an internal skeleton will keep the tissue straight without
affecting the epithelium. But maybe this can also result to artifacts due to
epithelial traction, I don't really know...

If you find some solution to the problem, please let us know.

Best wishes

yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

eikonika-at-otenet.gr
yorgosnikas-at-hotmail.com
Tel/fax +30 210 8957677
Mobile +30 6945 107477



El 19/02/10 20:44, "marie.cantino-at-uconn.edu" {marie.cantino-at-uconn.edu}
escribió:

}
} I was reminded by a recent posting (flattening samples for embedding)
} of a problem we have recently encountered.
}
} A student is trying to do SEM on the distal ends of critical point
} dried hummingbird tongues. The main body of the tongue is about a mm
} wide by 1-2 cm long, and bifurcates towards the end into two flat
} ribbons of tissue that form spirals during critical point drying, even
} if they are kept flat during the fixation and dehydration. We would
} like to force them to stay flat all the way through the critical point
} drying. Any sheets of material used to sandwich them would have to be
} rigid enough to prevent curling (possibly by mounting in a frame) but
} not mechanically damage the surface, and it would need to be stable
} chemically and allow adequate exchange during critical point drying.
} I would be interested in any experiences you have with different
} materials for creating such a sandwich (e.g. metal or polymer meshes,
} filter papers). I am also open to other ideas, but would prefer not
} to pin them, as it is likely to damage critical parts.
}
} Our protocol involves aldehyde and osmium fixation, followed by
} dehydration to 100% ethanol and critical point drying. No other
} solvents (besides the CO2) are used.
}
} Marie
}
} Dr. Marie E. Cantino
} Associate Professor of Physiology and Neurobiology
} Director, Electron Microscopy Laboratory
} University of Connecticut, Unit 3242
} Phone: 860-486-3588
} Fax: 860-486-6369
}
}








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From: tina-at-pbrc.hawaii.edu
Date: Mon, 22 Feb 2010 13:26:26 -0600
Subject: [Microscopy] Re: SEM: flattening tissue samples for CPD

Contents Retrieved from Microscopy Listserver Archives
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I apologize if I blinked and someone recommended this before, but how
about making a replica of the tongue surface?

Aloha from sunny and warm Hawaii, the only state with no hummingbirds,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************




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From: marie.cantino-at-uconn.edu
Date: Tue, 23 Feb 2010 13:57:27 -0600
Subject: [Microscopy] Re: SEM: flattening tissue samples for CPD

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

Here is a brief synopsis of suggestions, some of which did not appear
on the listserver, for flattening our hummingbird tongues

- make a surface replica
- use HMDS
- use cryoSEM or ESEM
- pin or wire the sample to a steel mesh rack at intervals along its
length
- make partial horizontal cuts perpendicular to the long axis to
relieve stress during drying, then realign just after CPD when the
tissue is still elastic
- insert a minuten pin through the tissue along the long axis of the
tongue
- cut into pieces, then reassemble on the stub after drying
- process in tiny envelopes or sandwiches made from polycarbonate
filters, filter paper, kimwipes, formvar, window screen or biopsy bag
material that has been stapled or sewn together

We hope to test at least a couple of these ideas. Thanks to all who
responded!

Marie


Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369


==============================Original Headers==============================
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From: edelmare-at-muohio.edu
Date: Wed, 24 Feb 2010 07:38:26 -0600
Subject: [Microscopy] Re: SEM: flattening tissue samples for CPD

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

Marie, et al.

Not to add to the confusion, but I have been thinking about this for a few days and what bothers me is that the tongues do not curl during fixation and solvent dehydration, but do curl following CO2 CPD. This suggests to me that that the CPD processing is at fault, and when I have seen similar things in my lab I always suspect improper CPD processing. My best test samples have always been 2% agar blocks (i.e. 2% agar 98% water/solvent/empty space) - if they come out of the CPD changed from when they went in then the CPD was not done right.

I have had aldehyde fixation and solvent dehydration all produce structural stresses and changes most typically and causing curling etc. in tissues. Because of differential size changes fixation rates structural water removal etc.

Problems with CPD: water left in tissue - incomplete dehydration. Failure to completely flush the solvent with CO2. Water in the CO2 (I think as has been discussed on this list this is very rare). Failure to reach critical point. BUT most common I have found is failure to leave a gaseous head space following the last CO2 flush. You need to leave a 20-30% (volume) head space in the CPD chamber. So you have a cold chamber (I really like 4-5 C) with 70-80% filled with liquid CO2 and 30-20% gaseous CO2, then isolate the chamber and raise the temp (which will raise the pressure) until critical point is reached (or passed). And then vent VERY slowly.


Richard E. Edelmann, Ph.D.
Electron Microscopy Facility Director
Miami University
Oxford, OH 45056

Ph: 513.529.5712
www.emf.muohio.edu

==================================
________________________________________
X-from: marie.cantino-at-uconn.edu [marie.cantino-at-uconn.edu]
Sent: Tuesday, February 23, 2010 2:59 PM
To: Edelmann, Richard E. Dr.

Here is a brief synopsis of suggestions, some of which did not appear
on the listserver, for flattening our hummingbird tongues

- make a surface replica
- use HMDS
- use cryoSEM or ESEM
- pin or wire the sample to a steel mesh rack at intervals along its
length
- make partial horizontal cuts perpendicular to the long axis to
relieve stress during drying, then realign just after CPD when the
tissue is still elastic
- insert a minuten pin through the tissue along the long axis of the
tongue
- cut into pieces, then reassemble on the stub after drying
- process in tiny envelopes or sandwiches made from polycarbonate
filters, filter paper, kimwipes, formvar, window screen or biopsy bag
material that has been stapled or sewn together

We hope to test at least a couple of these ideas. Thanks to all who
responded!

Marie


Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369




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From: ian.drucker-at-gmail.com
Date: Wed, 24 Feb 2010 11:15:57 -0600
Subject: [Microscopy] Image Colorizing

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

I'm looking for a Free Software program that can take SEM/FIB TIF images and
colorize them at designated areas. Any ideas? I've tried the usual free
image programs but so far I've found none that can easily do it.

Thanks

==============================Original Headers==============================
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2, 29 -- Message-ID: {c0bfda951002240915ue474891uc95fe871208e48ec-at-mail.gmail.com}
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==============================End of - Headers==============================




From: stefan.diller-at-t-online.de
Date: Wed, 24 Feb 2010 11:37:08 -0600
Subject: [Microscopy] Re: Image Colorizing

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

Dear Ian,
there is no easy way to get precisely coloured images. Best would be to use various detector-signals grabbed at the same time (to
get rid of pixel-offset,...) and blend them over via layers and / or paths in Photoshop or any other software able to handle image
data.

Best wishes,
Stefan


ian.drucker-at-gmail.com schrieb:
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} I'm looking for a Free Software program that can take SEM/FIB TIF images and
} colorize them at designated areas. Any ideas? I've tried the usual free
} image programs but so far I've found none that can easily do it.
}
} Thanks
}
} ==============================Original Headers==============================
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} ==============================End of - Headers==============================
}

--
-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

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From: edelmare-at-muohio.edu
Date: Wed, 24 Feb 2010 12:12:02 -0600
Subject: [Microscopy] Re: SEM: flattening tissue samples for CPD

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

Since a few folks have asked me "Why the headspace?" I thought I'd answer.

I am not sure as to why it works. We discovered it when I was a grad student. We were having problems with dehydration artifacts following CPD and one of us went back to the original early 1970's CPD paper and they very specifically said that the head space was needed for it to work. We tried it and everything was good! Lets here it for word of mouth learning vs going back to the source.

Sorry do not have the reference - but I bet someone else on the list does.

Richard E. Edelmann, Ph.D.
Electron Microscopy Facility Director
Miami University
Oxford, OH 45056

Ph: 513.529.5712
www.emf.muohio.edu
________________________________________
X-from: Louis Kerr [lkerr-at-mbl.edu]
Sent: Wednesday, February 24, 2010 11:42 AM
To: Edelmann, Richard E. Dr.

Marie, et al.

Not to add to the confusion, but I have been thinking about this for a few days and what bothers me is that the tongues do not curl during fixation and solvent dehydration, but do curl following CO2 CPD. This suggests to me that that the CPD processing is at fault, and when I have seen similar things in my lab I always suspect improper CPD processing. My best test samples have always been 2% agar blocks (i.e. 2% agar 98% water/solvent/empty space) - if they come out of the CPD changed from when they went in then the CPD was not done right.

I have had aldehyde fixation and solvent dehydration all produce structural stresses and changes most typically and causing curling etc. in tissues. Because of differential size changes fixation rates structural water removal etc.

Problems with CPD: water left in tissue - incomplete dehydration. Failure to completely flush the solvent with CO2. Water in the CO2 (I think as has been discussed on this list this is very rare). Failure to reach critical point. BUT most common I have found is failure to leave a gaseous head space following the last CO2 flush. You need to leave a 20-30% (volume) head space in the CPD chamber. So you have a cold chamber (I really like 4-5 C) with 70-80% filled with liquid CO2 and 30-20% gaseous CO2, then isolate the chamber and raise the temp (which will raise the pressure) until critical point is reached (or passed). And then vent VERY slowly.


Richard E. Edelmann, Ph.D.
Electron Microscopy Facility Director
Miami University
Oxford, OH 45056

Ph: 513.529.5712
www.emf.muohio.edu

==================================
________________________________________
X-from: marie.cantino-at-uconn.edu [marie.cantino-at-uconn.edu]
Sent: Tuesday, February 23, 2010 2:59 PM
To: Edelmann, Richard E. Dr.

Here is a brief synopsis of suggestions, some of which did not appear
on the listserver, for flattening our hummingbird tongues

- make a surface replica
- use HMDS
- use cryoSEM or ESEM
- pin or wire the sample to a steel mesh rack at intervals along its
length
- make partial horizontal cuts perpendicular to the long axis to
relieve stress during drying, then realign just after CPD when the
tissue is still elastic
- insert a minuten pin through the tissue along the long axis of the
tongue
- cut into pieces, then reassemble on the stub after drying
- process in tiny envelopes or sandwiches made from polycarbonate
filters, filter paper, kimwipes, formvar, window screen or biopsy bag
material that has been stapled or sewn together

We hope to test at least a couple of these ideas. Thanks to all who
responded!

Marie


Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369



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From: guenter.resch-at-imba.oeaw.ac.at
Date: Wed, 24 Feb 2010 13:24:27 -0600
Subject: [Microscopy] Re: Image Colorizing

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

Ian,

} I'm looking for a Free Software program that can take SEM/FIB TIF
} images and colorize them at designated areas. Any ideas? I've tried
} the usual free image programs but so far I've found none that can
} easily do it.

I don't know how much of automation you need, but The Gimp
(http://www.gimp.org/) is doing quite a good job in image manipulation
and it's open source.

Best,

Guenter

--
Dr. Guenter Resch, IMP-IMBA-GMI Electron Microscopy Facility
Email: guenter.resch-at-imp.ac.at, guenter.resch-at-imba.oeaw.ac.at
Phone: +43 (1) 79044-4250; Fax/Voice Mail: +43 (1) 79044-224250
Post: Dr. Bohr-Gasse 3 - 1030 Vienna - Austria, European Union

==============================Original Headers==============================
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From: kjmorris-at-well.ox.ac.uk
Date: Thu, 25 Feb 2010 04:08:59 -0600
Subject: [Microscopy] Re: Image Colorizing

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

Hi Ian,

Stefan is right, and it really is pretty easy, if time consuming, to do in
Photoshop CS4 [CS4 Extended's only £130 with Educational discounts - on a
really tight budget the likes Serif PhotoPlus X3 or Adobe Elements 8 will
suffice].

There's always different ways to approach something like this in Photoshop
CS4 [or earlier versions], but I'd try:

Load the SEM Photo and manually select the region you wish to colourize,
using say the 'quick selection tool' - you can add/reject [shift/alt] bits
of selected regions with a lasso tool. Then ensure the image is RGB colour
[image, mode]. I'd then go to 'image, adjustments, colour balance' and
lightly adjust the colour balance of the selected region and then "you
could be brown, you could be blue, you could be violet sky", e.g. move the
colour balance slider from cyan to red and the selected object with become
progressively more red, with the underlying structural details intact.
Then work through the entire image. Select all objects of one colour as
one mutliple region for colourizing [shift], and you can say slightly
adjust contrast and brightness, or curves, or shadow/highlights within
those selected regions as well.

You don't really need layers, you could work on the main image
[background] bit by bit - save regularly under new file names as
Photoshops undo [step backwards] is limited if, after a lot of work, you
don't like the way the image is turning out.

You could do all this with Photoshop Elements 8 as well [£37 with
discounts]. Do all the above Photoshop stuff using the similar Elements
tools, then [instead of 'colour balance'] goto: Enhance, Adjust Colour,
Adjust Hue/Saturation [and make sure the 'Colorize' box is ticked].

It will take a while to manually edit the entire image ['View, Zoom' to
aid tracing], but I doubt any image processing software could fully
'automatically' select the regions you want to colorize, particularly with
a '3D' SEM image. Photoshop's 'Quick selection tool' will have a go though
[the tools selection effect is adjustable in the upper menu bar].

I doubt the likes of 'Pseudocolour' LUT effects will provide the subtle
colouring you require, although they may work adaquately on TEM images of
sections.

Regards

Keith

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

Dr Keith J Morris
Molecular Cytogenetics and Microscopy Core
The Wellcome Trust Centre for Human Genetics
Roosevelt Drive
Oxford
OX3 7BN
United Kingdom

Tel: +44 ( 0 ) 1865 287568
Email: kjmorris-at-well.ox.ac.uk
HomePage: http://www.well.ox.ac.uk/cytogenetics





}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} ----------------------------------------------------------------------------
}
} Dear Ian,
} there is no easy way to get precisely coloured images. Best would be to
} use various detector-signals grabbed at the same time (to
} get rid of pixel-offset,...) and blend them over via layers and / or paths
} in Photoshop or any other software able to handle image
} data.
}
} Best wishes,
} Stefan
}
}
} ian.drucker-at-gmail.com schrieb:
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} } America
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} } http://www.microscopy.com/MicroscopyListserver
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} } ----------------------------------------------------------------------------
} }
} } I'm looking for a Free Software program that can take SEM/FIB TIF images
} } and
} } colorize them at designated areas. Any ideas? I've tried the usual
} } free
} } image programs but so far I've found none that can easily do it.
} }
} } Thanks
} }
} } ==============================Original
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} }
}
} --
} -----------------------------------------------------
} Stefan Diller - Scientific Photography
} Arndtstrasse 22
} D - 97072 Wuerzburg Germany
} ++49-931-7848700 Phone
} ++49-931-7848701 Fax
} ++49-175-7177051 Mobile
}
} Websites:
} www.stefan-diller.com
} www.elektronenmikroskopie.info
} www.assisi.de
} www.zwillingsprojekt.de
} Anfahrt: http://Mail.map24.com/Stefan.Diller
} -----------------------------------------------------
}
} ==============================Original
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From: PhillipsT-at-missouri.edu
Date: Thu, 25 Feb 2010 08:12:19 -0600
Subject: [Microscopy] LM - lubrication problem

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

I have inherited two compound microscopes with similar problems. The focusing oculars of one are very tight so the rotating mechanism needs to be cleaned and re-lubricated. In the other scope, it is the condenser field diaphragm that is tight. Any ideas on how to approach this in regards to cleaning and what lubrication? I am, of course, aware that there are professionals who can be hired but would like to do these two scopes on my own.


Thomas E. Phillips, Ph.D.
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
Biological Sciences
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (voice)
573-882-0123 (fax)




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From: bozzola-at-siu.edu
Date: Thu, 25 Feb 2010 09:20:07 -0600
Subject: [Microscopy] Re: LM - lubrication problem

Contents Retrieved from Microscopy Listserver Archives
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Tom,

This problem is usually caused by polymerization (or drying out) of
the lubricant, resulting in metal upon metal rubbing of the moving
parts.

You need to disassemble the part, taking care to remove (if possible)
the optics safely away from the metal parts. Then, try soaking the
metal parts in xylene to loosen or dissolve the grease. Then, clean
the parts with cotton swabs or lintless cloth (soaked in xylene). You
may have to repeat this soaking and rubbing many times to get through
the built up grease.

When sufficiently clean, the parts should move smoothly against each
other, even without lube. Finally, apply a very thin layer of lithium
grease on the moving parts.

I've cleaned many microscopes over the years and failed to get only a
couple parts working again (a microscope focusing stage and a
condenser) since I was unable to disassemble them.

John Bozzola
Carbondale, IL

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From: bigelow-at-umich.edu
Date: Thu, 25 Feb 2010 13:22:35 -0600
Subject: [Microscopy] RE: Lubricant for microscopes

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

The perfluorinated polyphenyl ether oils (Fomblin and Krytox) and
greases (Brayco and Krytox) are superb lubricants, AND they are
chemically inert and do not oxidize or polymerize. They also have
very low vapor pressures (10-10 Torr or better) and can therefore be
used as lubricants for things inside a vacuum system.

As I recall, the guy that services microscopes here at the U of Mich.
has been using these very successfully for a number of years
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From: ALawrence-at-entomology.msstate.edu
Date: Thu, 25 Feb 2010 15:56:52 -0600
Subject: [Microscopy] postdoctoral position

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


A Post Doctoral Fellow position has been made possible through an NSF
MRI 2009 award for atomic force microscopy (AFM) instrumentation. The
post doc will reside at Mississippi State University's Electron
Microscope Center, will have a primary responsibility for the operation,
maintenance, and training for the new AFM instruments, and will be
actively involved in independent as well as collaborative research
efforts. We appreciate your help in identifying potential candidates
with AFM expertise. Please distribute the information below widely as
you see fit. Please feel free to contact giselle-at-emcenter.msstate.edu
with questions.

Post Doctoral Research Position in
Atomic Force/Scanning Probe Microscopy available immediately

Mississippi State University (MSU) has an opening for a Postdoctoral
Research Associate with expertise in atomic force and scanning probe
microscopy (AFM/SPM) techniques. This postdoc will work with a diverse,
dynamic team of materials and life science investigators in MSU*s
Electron Microscope Center (EMC), a university-wide core facility with
missions in research, teaching, and service.

The successful applicant must have a Ph.D. (or equivalent degree) in an
engineering or physical/natural science field and a strong background in
AFM/SPM theory and operation. Knowledge of other general imaging
approaches is desired. The applicant must have a demonstrated ability to
work independently, to work in a collaborative environment with a
diverse group of colleagues, and to communicate effectively in written
and oral formats.

The successful applicant will be responsible for operation,
maintenance, and user training for new state-of-the-art atomic force
microscopy instruments within the multi-user EMC. The applicant will
primarily support the AFM instrumentation, but will also have the
opportunity to learn/operate other imaging and characterization
technologies available at the EMC. In terms of professional
development, the applicant will be encouraged to develop an independent
research program, contribute to interdisciplinary research efforts, and
direct graduate and undergraduate student research assistants.

Review will begin immediately and will continue until the position is
filled. Applicants should complete an online application at
www.jobs.msstate.edu/applicants/Central?quickFind=178227 and upload
a cover letter and current vitae. Applicants should also submit three
letters of recommendation (mailed under separate cover) and a transcript
from their terminal degree program. Letters of recommendation and
transcripts should be mailed to Sherre Denson, Business Manager, Dave C.
Swalm School of Chemical Engineering, Mississippi State University, Box
9595, Mississippi State, MS 39762 (sherre-at-che.msstate.edu).

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From: paul.gerroir-at-xrcc.xeroxlabs.com
Date: Thu, 25 Feb 2010 16:14:17 -0600
Subject: [Microscopy] TEM - Embedding Resins

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

All,
I am exploring potential alternatives to the two-part epoxy
(resin/hardener) that we currently use. Does anyone have any experience
with materials that can be safely used for encapsulating polyester
particles and fibres without the concern for infiltration or chemical
reaction? Most of the two-part epoxies we have been using thus far are
bisphenol-based and cure at room temperature. UV curing is a possibility
provided the temperature is kept below 50C.

Thanks,
Paul

Paul J. Gerroir

Microscopy
Materials Characterization
Xerox Research Centre of Canada
2660 Speakman Drive
Mississauga, Ontario L5K 2L1

Phone: 905-823-7091, ext.216
FAX: 905-822-7022
e-mail: paul.gerroir-at-xerox.com



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From: TindallR-at-missouri.edu
Date: Thu, 25 Feb 2010 16:52:42 -0600
Subject: [Microscopy] Immunocyto of insulin: skeletal muscle

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

Dear Collective,

We have a client who is desperately trying to localize insulin in skeletal muscle at the TEM level. So far, not so good, but we're still trying. We are going to work with positive controls to test our primaries and give it another whirl(s), but we are looking for any suggestions ya'll might have. Our literature search has been negative, so far. We find studies which label insulin in the pancreas and studies which label Insulin-like Growth Factors, but not insulin directly in the muscle.

The antibody we tried first was successful in LM.

Thanks in advance.

Cheers,
Randy


Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




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From: gary-at-gaugler.com
Date: Thu, 25 Feb 2010 17:08:06 -0600
Subject: [Microscopy] Re: Image Colorizing

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

There aren't any free programs that I am aware of for
pseudo colorization. The paid programs will do the
whole image but not selected areas. If you use Photoshop,
you can colorize selected areas and as different colors.

gary g.


At 09:19 AM 2/24/2010, you wrote:
} ----------------------------------------------------------------------------
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From: stevem-at-vt.edu
Date: Thu, 25 Feb 2010 19:10:50 -0600
Subject: [Microscopy] viaWWW: roughness measurments in the SEM

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Email: stevem-at-vt.edu
Name: Steve McCartney

Organization: Va Tech

Title-Subject: [Filtered] roughness measurments in the SEM

Message: Hello: We are trying to get surface roughness measurements
on sample that has height variations from about 50nm to several
microns. We were not able to use AFM. I have read a little about
the software that can be used to measure roughness in the SEM. I am
wondering if this can be used to measure such extreme differences in
roughness and if so does anyone have any recommendations as to which
software to use. Also are there other techniques we should consider.
Thanks in advance for any help. Steve McCartney

Login Host: 198.82.148.215
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From: edelmare-at-muohio.edu
Date: Fri, 26 Feb 2010 02:34:24 -0600
Subject: [Microscopy] TEM - Embedding Resins

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

Paul:

Sorry, not much of an organic chemist. There are a number of "cold" UV curing resins - cold = 0 to -20 to -60 C which are used for Biological TEM prep. Biological embedding resins tend to be multi-component resins, but some of the cold curing are one to two part. Like http://www.emsdiasum.com/microscopy/products/embedding/kits.aspx#14330

I do not have any specific recommendations but you might try looking in that direction.

I know that things like Sulfur have been used for embedding as well but its melting point is 107C a little too warm for you.

If you go with the biol resins avoid the methacylates they are often exothermic during polymerization and get far above 70C in micro-regions



Richard E. Edelmann, Ph.D.
Electron Microscopy Facility Director
Miami University
Oxford, OH 45056

Ph: 513.529.5712
www.emf.muohio.edu
________________________________________
X-from: paul.gerroir-at-xrcc.xeroxlabs.com [paul.gerroir-at-xrcc.xeroxlabs.com]
Sent: Thursday, February 25, 2010 5:14 PM
To: Edelmann, Richard E. Dr.

All,
I am exploring potential alternatives to the two-part epoxy
(resin/hardener) that we currently use. Does anyone have any experience
with materials that can be safely used for encapsulating polyester
particles and fibres without the concern for infiltration or chemical
reaction? Most of the two-part epoxies we have been using thus far are
bisphenol-based and cure at room temperature. UV curing is a possibility
provided the temperature is kept below 50C.

Thanks,
Paul

Paul J. Gerroir

Microscopy
Materials Characterization
Xerox Research Centre of Canada
2660 Speakman Drive
Mississauga, Ontario L5K 2L1

Phone: 905-823-7091, ext.216
FAX: 905-822-7022
e-mail: paul.gerroir-at-xerox.com



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From: kjmorris-at-well.ox.ac.uk
Date: Fri, 26 Feb 2010 05:12:07 -0600
Subject: [Microscopy] Re: Image Colorizing

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

Hi Ian,

Tried out colorizing an SEM image with Photoshop CS4 and the results seemed
fine:

Original SEM image of pollen
http://en.wikipedia.org/wiki/File:Misc_pollen.jpg

I Colorized the image using the method in my last post [you could use
Photoshop CS4 or Elements 8]
http://www.well.ox.ac.uk/cytogenetics/pollen/pollen_colorized.jpg
It took well under an hour to select and colorize the whole image [and I
found it quite therapeutic].

I also applied a Pseudocolor look-up table [LUT] applied using Photoshop CS4
[and Elements 8 has similar tools]
http://www.well.ox.ac.uk/cytogenetics/pollen/pollen_pseudo.jpg
See details of using pseudocolor LUTs with Photoshop see the likes of
http://www.rawlight.com/psuedo.pdf
and search the internet


I have seen one free image editor that apparently can colorize images fairly
well [it's used by artists, and some of the results are pretty stunning, if
a little gaudy]
But the editor interface seems very clunky and I'm not sure how to upload
and save the images using it. It talks of your edited images being available
for all, i.e. you might lose copyright or at least your images might be free
for others to view or steal.

http://aviary.com
http://www.well.ox.ac.uk/cytogenetics/pollen/pollen_dayglo.jpg
http://fxh.worth1000.com/entries/507038
http://lifehacker.com/5309162/best-online-image-editor-aviary-phoenix

But it is apparently free to use [might need Internet explorer].

However any opportunity to hone your Photoshop skills is probably worth
taking.

Regards

Keith

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford  OX3 7BN,
United Kingdom.

Telephone:  +44 (0)1865 287568
Email:  kjmorris-at-well.ox.ac.uk
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy

-----Original Message-----
X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com]
Sent: 25 February 2010 23:14
To: kjmorris-at-well.ox.ac.uk

There aren't any free programs that I am aware of for
pseudo colorization. The paid programs will do the
whole image but not selected areas. If you use Photoshop,
you can colorize selected areas and as different colors.

gary g.


At 09:19 AM 2/24/2010, you wrote:
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From: ian.drucker-at-gmail.com
Date: Fri, 26 Feb 2010 08:25:46 -0600
Subject: [Microscopy] Re: Image Colorizing

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

Thank you all for the info. It's clear that I'd be best off with
Photoshop but since I don't have that option at the moment I'l try
GIMP and see how that goes.

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From: DusevichV-at-umkc.edu
Date: Fri, 26 Feb 2010 09:03:23 -0600
Subject: [Microscopy] RE: viaWWW: roughness measurements in the SEM

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

You can take a look at MeX from Alicona. It was created for SEM and can
measure roughness.
http://www.alicona.com/englisch/main-navigation/products/mex/index.html


Vladimir
(Happy user, but I do not measure roughness...)

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

} -----Original Message-----
} From: stevem-at-vt.edu [mailto:stevem-at-vt.edu]
} Sent: Thursday, February 25, 2010 7:11 PM
} To: Dusevich, Vladimir
} Subject: [Microscopy] viaWWW: roughness measurments in the SEM
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} Email: stevem-at-vt.edu
} Name: Steve McCartney
}
} Organization: Va Tech
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} Title-Subject: [Filtered] roughness measurments in the SEM
}
} Message: Hello: We are trying to get surface roughness measurements
} on sample that has height variations from about 50nm to several
} microns. We were not able to use AFM. I have read a little about
} the software that can be used to measure roughness in the SEM. I am
} wondering if this can be used to measure such extreme differences in
} roughness and if so does anyone have any recommendations as to which
} software to use. Also are there other techniques we should consider.
} Thanks in advance for any help. Steve McCartney
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From: mdufraine-at-ebsciences.com
Date: Fri, 26 Feb 2010 09:29:43 -0600
Subject: [Microscopy] viaWWW: roughness measurments in the SEM

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

Steve-

Energy Beam Sciences is the US distributor, for the Alicona MeX 5.1 3D Surface Metrology Software System for SEM images. MeX provides for Profile, Area, volume, and 2D Image Analysis, with roughness measurements and parameters, one of the highlighted functions of the software.
I will provide to you, off-line, information on this unique, and easy to use 3D measurement software.

Mike Dufraine
Energy Beam Sciences, Inc.

-----Original Message-----
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Email: stevem-at-vt.edu
Name: Steve McCartney

Organization: Va Tech

Title-Subject: [Filtered] roughness measurments in the SEM

Message: Hello: We are trying to get surface roughness measurements
on sample that has height variations from about 50nm to several
microns. We were not able to use AFM. I have read a little about
the software that can be used to measure roughness in the SEM. I am
wondering if this can be used to measure such extreme differences in
roughness and if so does anyone have any recommendations as to which
software to use. Also are there other techniques we should consider.
Thanks in advance for any help. Steve McCartney

Login Host: 198.82.148.215
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From: protrain-at-emcourses.com
Date: Fri, 26 Feb 2010 10:07:55 -0600
Subject: [Microscopy] viaWWW: roughness measurments in the SEM

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

Hi

I am always fascinated when people consider measuring surface roughness. I
have had a mass of questions over the past few years on this subject due to
the "developments" in anti wrinkle treatments; how rough is the surface?

Why I am fascinated is that as far as I am concerned the roughness of a
surface may be "modified" through the use of different imaging techniques.
Crudely, to use a high kV and a surface looks rougher, use a low kV and it
looks smooth, it all depends upon the surface/sub surface relationship and
penetration.

An example - using rough emery paper grind flat the surface of an aluminium
plate. Leave this for a day or two and then examine it in the SEM. At low
kV it will probably charge and, even when coated, look smooth. At high kV
you will penetrate through the oxide film and see the original rough
surface.

What is the general view on surface roughness monitoring, or is this so
gross in SEM terms that the instrument's interpretation of the image is not
considered to be a problem.

I am interested in comment?


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide Tel +44
(0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com


-----Original Message-----
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To: protrain-at-emcourses.com

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Email: stevem-at-vt.edu
Name: Steve McCartney

Organization: Va Tech

Title-Subject: [Filtered] roughness measurments in the SEM

Message: Hello: We are trying to get surface roughness measurements
on sample that has height variations from about 50nm to several
microns. We were not able to use AFM. I have read a little about
the software that can be used to measure roughness in the SEM. I am
wondering if this can be used to measure such extreme differences in
roughness and if so does anyone have any recommendations as to which
software to use. Also are there other techniques we should consider.
Thanks in advance for any help. Steve McCartney

Login Host: 198.82.148.215
---------------------------------------------------------------------------

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From: donc-at-asmicro.com
Date: Fri, 26 Feb 2010 12:11:00 -0600
Subject: [Microscopy] roughness measurments

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

Dear Steve,
You asked about surface roughness measurements over a wide range of
roughnesses. You implied that you tried AFM and failed. If so, it could be
due to many causes, including specific limitations of the AFM you tried. An
AFM with a larger Z axis range may be more suitable.

The AFM equipment at Advanced Surface Microscopy (ASM) can measure surface
features from 0.1 nm to 25 um high, so perhaps we can help you. We
routinely measure pit depths of 10 um and higher.
Please contact me offline for more details.

I would like to comment on other messages in this thread that relate to
choice of techniques.

1) AFM is one of the best ways to measure surface roughness, since the
instrument is designed to provide quantitative measurements on the Z axis.
2)Another technique to consider is optical interference microscopy, using
instruments made by Wyko or Zygo, which also are designed to provide
quantitative height information.
3) Although SEM software add-ons may yield Z axis measurements, I have not
seen any publications that show the accuracy of those measurements, e.g. by
using software to measure step heights on surface topography reference
specimens. If there are such publications, please let me know.

Disclaimer: ASM is a commercial analytical service lab and a used equipment
dealer specializing in AFM. We have a financial interest in promoting the
productive use of AFM to solve problems.

regards,
Don
=============================================
"Can you calibrate a 1 Million X SEM or AFM image? Ask us how."

Don Chernoff, Ph.D., President
Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com
3250 N. Post Rd., Ste. 120 Voice: 317-895-5630
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From: stevem-at-vt.edu
To: donc-at-asmicro.com
Sent: Thursday, February 25, 2010 8:13 PM
Subject: [a] [Microscopy] viaWWW: roughness measurments in the SEM





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Email: stevem-at-vt.edu
Name: Steve McCartney

Organization: Va Tech

Title-Subject: [Filtered] roughness measurments in the SEM

Message: Hello: We are trying to get surface roughness measurements
on sample that has height variations from about 50nm to several
microns. We were not able to use AFM. I have read a little about
the software that can be used to measure roughness in the SEM. I am
wondering if this can be used to measure such extreme differences in
roughness and if so does anyone have any recommendations as to which
software to use. Also are there other techniques we should consider.
Thanks in advance for any help. Steve McCartney

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From: jquinn-at-www.matscieng.sunysb.edu
Date: Fri, 26 Feb 2010 13:03:14 -0600
Subject: [Microscopy] re: roughness by SEM

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


Folks (in and out of office) -

While not completely skeptical of photogrammametric
techniques and SEM imaging for surface roughness analysis,
one must question its use with parameters that make objects
appear and vanish (detector type, KV/HT, tilt, WD, sample
morphology/chemistry, coating/charging, non-uniform
image background, etc.....).

regards,

JQuinn


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From: bob.price-at-uscmed.sc.edu
Date: Fri, 26 Feb 2010 18:25:58 -0600
Subject: [Microscopy] viaWWW: Basic Confocal Microscopy Workshop

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Email: bob.price-at-uscmed.sc.edu
Name: Bob Price

Organization: Univ South Carolina

Title-Subject: [Filtered] Confocal Workshop

Message: The University of South Carolina School of Medicine
Instrumentation Resource Facility will be hosting the 6th Annual
Workshop on Basic Confocal Microscopy from June 14th through the
18th. The workshop is directed towards beginning and intermediate
users of confocal microscopes and involves a series of lectures
(specimen preparation, labeling strategies, proper set-up of
instrument operating parameters, proper handling of 2D and 3D
confocal images in programs such as Adobe Photoshop and AMIRA), hands
on specimen preparation, and time on a number of different point
scanning and spinning disk confocal microscopes.

Companies scheduled to have instruments and applications experts
on-site include BioVision Technologies, Leica, Nikon, Olympus, Perkin
Elmer and Zeiss.

Faculty will include Dr. Jay Jerome of Vanderbilt University, Dr.
Ralph Albrecht of the University of Wisconsin Madison, Dr. John
Mackenzie of North Carolina State University, Dr. Tom Trusk of the
Medical University of South Carolina, and Dr. Bob Price of the
University of South Carolina.

For further information please see:
http://dba.med.sc.edu/price/irf/irf.htm or contact Anna McNeal
(anna.mcneal-at-uscmed.sc.edu) or Bob Price (bob.price-at-uscmed.sc.edu).


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From: CGoldsmith-at-cdc.gov
Date: Sat, 27 Feb 2010 10:00:30 -0600
Subject: [Microscopy] viaWWW: SEMS annual meeting May 24-26, 2010

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Email: a.kushch-at-hi-z.com
Name: Aleks Kushch

Organization: Hi-Z Technology

Title-Subject: [Filtered] TEM and EBIC capability

Message: We are looking for a place that has a TEM with EBIC capability

Sincerely
Aleks

Aleksandr (Aleks) S. Kushch, Ph.D
Program Manager
Hi-Z Technology, Inc.
7606 Miramar Road
San Diego, CA 92126
Telephone: (858) 695-6660
Fax: (858) 695-8870
e-mail: a.kushch-at-hi-z.com
http://www.hi-z.com


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Email: CGoldsmith-at-cdc.gov
Name: Cynthia Goldsmith

Organization: Southeastern Microscopy
Society/Centers for Disease Control and Prevention

Title-Subject: [Filtered] SEMS annual meeting May 24-26, 2010

Message: On behalf of the Southeastern Microscopy
Society (SEMS) Local Arrangements and Program
Committees, I would like to invite you to the
SEMS annual meeting which will be held from May
24-26, 2010 at the Francis Marion hotel in
historical Charleston, South Carolina.

Microscopy Society of America, Microbeam Analysis
Society, and SEMS sponsored speakers include:
Dr. David Piston (MSA President) ñ Vanderbilt University
Dr. David Joy ñ University of Tennessee
Dr. Paul Kotula ñ Sandia National Laboratories
Dr. Michael Postek ñ NIST
Dr. Elaine Schumacher ñ McCrone Associates, Inc
Dr. Niels de Jonge - Vanderbilt University Medical Center
Dr. John Lemasters ñ Medical University of South Carolina
Dr. Sue Lessner ñ University of South Carolina
Dr. Heather Evans-Anderson ñ Winthrop University
Dr. Anand Ramamurthi ñ Clemson University

Assistance for travel and meeting costs for
students competing in the SEMS Ruska Award and
technologists is available. There are also
opportunities to participate in pre-meeting
workshops on Confocal Microscopy and Digital
Imaging, a tour of the Robert Bosch Company
Facility, and post-meeting tours of the Yorktown
and Fort Sumter.

For further information on abstract submission,
registration, and participation in the above
workshops and tours please visit:
{http://www.southeasternmicroscopy.org/index-2.html} http://www.southeasternmicroscopy.org/index-2.html
and
{http://www.southeasternmicroscopy.org/meetings_doc/SEMSBro_web.pdf} http://www.southeasternmicroscopy.org/meetings_doc/SEMSBro_web.pdf

The deadline for abstract submission is Thursday,
April 1, 2010. I hope all of you will be able to
attend.

Sincerely,
Robert L. Price
President, SEMS


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From: protrain-at-emcourses.com
Date: Sat, 27 Feb 2010 15:17:59 -0600
Subject: [Microscopy] viaWWW: roughness measurments in the SEM

Contents Retrieved from Microscopy Listserver Archives
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Hi Paul

That is exactly my point, change the kV change the information. Is there a
specification for investigating surface roughness that states that the
scientist should work at low kV; how low is low?

My experience is that people will work at the kV that demonstrates the most
attractive image of their material, is that the image that they will use to
examine roughness? Often the most attractive image contains a degree of sub
surface information which adds contrast, could that be used for surface
roughness investigations?

I work around the world with different cultures, different instruments and
many different materials. I am not convinced that the average SEM operator
would work at a kV that will only show the true specimen surface, it's not
in their nature; you are talking of {2kV, or even lower for some materials?

Steve

-----Original Message-----
X-from: Gerroir, Paul [mailto:paul.gerroir-at-xrcc.xeroxlabs.com]
Sent: 26 February 2010 20:19
To: protrain-at-emcourses.com

Hi

I am always fascinated when people consider measuring surface roughness.
I
have had a mass of questions over the past few years on this subject due
to
the "developments" in anti wrinkle treatments; how rough is the surface?

Why I am fascinated is that as far as I am concerned the roughness of a
surface may be "modified" through the use of different imaging
techniques.
Crudely, to use a high kV and a surface looks rougher, use a low kV and
it
looks smooth, it all depends upon the surface/sub surface relationship
and
penetration.

An example - using rough emery paper grind flat the surface of an
aluminium
plate. Leave this for a day or two and then examine it in the SEM. At
low
kV it will probably charge and, even when coated, look smooth. At high
kV
you will penetrate through the oxide film and see the original rough
surface.

What is the general view on surface roughness monitoring, or is this so
gross in SEM terms that the instrument's interpretation of the image is
not
considered to be a problem.

I am interested in comment?


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide Tel +44
(0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com


-----Original Message-----
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Sent: 26 February 2010 01:12
To: protrain-at-emcourses.com

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Email: stevem-at-vt.edu
Name: Steve McCartney

Organization: Va Tech

Title-Subject: [Filtered] roughness measurments in the SEM

Message: Hello: We are trying to get surface roughness measurements
on sample that has height variations from about 50nm to several
microns. We were not able to use AFM. I have read a little about
the software that can be used to measure roughness in the SEM. I am
wondering if this can be used to measure such extreme differences in
roughness and if so does anyone have any recommendations as to which
software to use. Also are there other techniques we should consider.
Thanks in advance for any help. Steve McCartney

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From: frah0010-at-umn.edu
Date: Sat, 27 Feb 2010 17:05:50 -0600
Subject: [Microscopy] Re: roughness measurements in the SEM

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

} ... I work around the world with different cultures, different
} instruments and many different materials. I am not convinced that
} the average SEM operator would work at a kV that will only show the
} true specimen surface, it's not in their nature...

Steve,

You've identified a topic of interest to anthropologists who study
technology: what are the non-technical factors that go into making
choices of how people use or understand technology? This can include
superstitions and what I call "lab culture" or "lab folklore" (i.e.,
bits of knowledge, correct or not, handed down from lab user to lab
user about how to do something or how it works). This is an interest
of mine as it relates to analytical techniques, since I have a
background in anthropology as well. Someday I'd like to essentially
do anthropological fieldwork in a series of laboratories to study
their different technological choices and identify the factors that
affect them. I've argued before, though, that based on my own
experience, ISO-type standards for analytical and microscopic
techniques leave plenty of room for non-technical choices to remain,
despite addressing purely technical details (like the instrument's
specifications) well. Just as important, how such standards, in turn,
actually affect an analyst, especially one who is already ingrained in
the "culture" of their lab and may have preconceptions that go
unchanged, just like students in a physics course come to the class
with an idea of how the world works and teaching that out of them is
hard, often requiring science education specialists to identify those
alternative views and how to correct them. The same evaluation needs
to be done with standard practices, that is, after the issues needed
to formulate them have been resolved. And let's not even get started
on the topic of theory-dependence!

Best,
Ellery

---------------
Ellery Frahm
Manager & Principal Analyst, Electron Microprobe Lab
Senior Research Fellow, Geology and Geophysics
Doctoral Candidate, Department of Anthropology
University of Minnesota - Twin Cities
http://web.mac.com/elleryfrahm/

==============================Original Headers==============================
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From: kraftpiano-at-gmail.com
Date: Sat, 27 Feb 2010 20:57:07 -0600
Subject: [Microscopy] roughness measurements in the SEM

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

I've been following this thread with interest, hoping to find out
exactly how one would measure "Roughness." It seems a little too
subjective to me to be quantified.

Reading this reply, however, made me remember a technician bI once
worked with who swore that the ICPMS would give her better results if
she gently rubbed the top of the machine. She was so sure of this
that in the operation instructions for that site, one of the steps
actually reads "Rub the top of the machine to ward off evil spirits
and get good data."

--Justin A. Kraft

On Sat, Feb 27, 2010 at 6:10 PM, {frah0010-at-umn.edu} wrote:
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} } ... I work around the world with different cultures, different
} } instruments and many different materials. I am not convinced that
} } the average SEM operator would work at a kV that will only show the
} } true specimen surface, it's not in their nature...
}
} Steve,
}
} You've identified a topic of interest to anthropologists who study
} technology: what are the non-technical factors that go into making
} choices of how people use or understand technology?  This can include
} superstitions and what I call "lab culture" or "lab folklore" (i.e.,
} bits of knowledge, correct or not, handed down from lab user to lab
} user about how to do something or how it works).  This is an interest
} of mine as it relates to analytical techniques, since I have a
} background in anthropology as well.  Someday I'd like to essentially
} do anthropological fieldwork in a series of laboratories to study
} their different technological choices and identify the factors that
} affect them.  I've argued before, though, that based on my own
} experience, ISO-type standards for analytical and microscopic
} techniques leave plenty of room for non-technical choices to remain,
} despite addressing purely technical details (like the instrument's
} specifications) well.  Just as important, how such standards, in turn,
} actually affect an analyst, especially one who is already ingrained in
} the "culture" of their lab and may have preconceptions that go
} unchanged, just like students in a physics course come to the class
} with an idea of how the world works and teaching that out of them is
} hard, often requiring science education specialists to identify those
} alternative views and how to correct them.  The same evaluation needs
} to be done with standard practices, that is, after the issues needed
} to formulate them have been resolved.  And let's not even get started
} on the topic of theory-dependence!
}
} Best,
} Ellery
}
} ---------------
} Ellery Frahm
} Manager & Principal Analyst, Electron Microprobe Lab
} Senior Research Fellow, Geology and Geophysics
} Doctoral Candidate, Department of Anthropology
} University of Minnesota - Twin Cities
} http://web.mac.com/elleryfrahm/
}
} ==============================Original Headers==============================
} 5, 23 -- From frah0010-at-umn.edu Sat Feb 27 17:05:50 2010
} 5, 23 -- Received: from vs-m.tc.umn.edu (vs-m.tc.umn.edu [134.84.135.97])
} 5, 23 --        by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id o1RN5oqU011673
} 5, 23 --        for {microscopy-at-microscopy.com} ; Sat, 27 Feb 2010 17:05:50 -0600
} 5, 23 -- Received: from [10.51.51.92] ([198.36.216.1])
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} 5, 23 -- Message-Id: {016F724D-2D3A-480F-89D5-65594878C41B-at-umn.edu}
} 5, 23 -- From: Ellery Frahm {frah0010-at-umn.edu}
} 5, 23 -- To: "protrain-at-emcourses.com" {protrain-at-emcourses.com} ,
} 5, 23 --         "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
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} 5, 23 -- Date: Sat, 27 Feb 2010 17:05:21 -0600
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} ==============================End of - Headers==============================



--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar


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8, 35 -- Subject: Re: [Microscopy] Re: roughness measurements in the SEM
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From: Frank_Karl-at-lincolnelectric.com
Date: Mon, 1 Mar 2010 06:23:05 -0600
Subject: [Microscopy] Re: roughness measurements in the SEM

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

At risk of being ex-communicated:

I once had a friend who need a toilet seat and decided to make one. He
bought the best lumber possible, straight grain lumber from far away lands,
the best paints and metal hardware. He shaped, cut and sanded the wood
followed by many coats of hand rubbed lacquer over a metallic gold flake
clear coat. The seat was installed with pride and great fanfare.

At the divorce his soon to be ex-wife testified she just want a new seat
that didn't give her slivers in her fanny.

He got custody of the seat.


I was going to launch a diatribe against this entire direction surface
roughness has taken, in which the operator seems to be focal point. Let me
remind you many operators are faced with restriction of what is available
and the requirements imposed by the organization owning the SEM. It might
be nice to decide to use gold coating at 2.5 kev, but if all you have is
carbon and you know EDS will be needed, well so much for surface. So
before we apply for that grant to study the cultural ties and social mores
of SEM operators maybe we should acknowledge that even with the knowledge
and skill set we are limited by the equipment and organizational goals
imposed on us.

Frank



kraftpiano-at-gmail.
com
To
02/27/2010 10:05 frank_karl-at-lincolnelectric.com
PM cc

Subject
Please respond to [Microscopy] roughness
kraftpiano-at-gmail. measurements in the SEM
com












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


I've been following this thread with interest, hoping to find out
exactly how one would measure "Roughness." It seems a little too
subjective to me to be quantified.

Reading this reply, however, made me remember a technician bI once
worked with who swore that the ICPMS would give her better results if
she gently rubbed the top of the machine. She was so sure of this
that in the operation instructions for that site, one of the steps
actually reads "Rub the top of the machine to ward off evil spirits
and get good data."

--Justin A. Kraft

On Sat, Feb 27, 2010 at 6:10 PM, {frah0010-at-umn.edu} wrote:
}
}
}
}
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} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of
America
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}
} } ... I work around the world with different cultures, different
} } instruments and many different materials. I am not convinced that
} } the average SEM operator would work at a kV that will only show the
} } true specimen surface, it's not in their nature...
}
} Steve,
}
} You've identified a topic of interest to anthropologists who study
} technology: what are the non-technical factors that go into making
} choices of how people use or understand technology?  This can include
} superstitions and what I call "lab culture" or "lab folklore" (i.e.,
} bits of knowledge, correct or not, handed down from lab user to lab
} user about how to do something or how it works).  This is an interest
} of mine as it relates to analytical techniques, since I have a
} background in anthropology as well.  Someday I'd like to essentially
} do anthropological fieldwork in a series of laboratories to study
} their different technological choices and identify the factors that
} affect them.  I've argued before, though, that based on my own
} experience, ISO-type standards for analytical and microscopic
} techniques leave plenty of room for non-technical choices to remain,
} despite addressing purely technical details (like the instrument's
} specifications) well.  Just as important, how such standards, in turn,
} actually affect an analyst, especially one who is already ingrained in
} the "culture" of their lab and may have preconceptions that go
} unchanged, just like students in a physics course come to the class
} with an idea of how the world works and teaching that out of them is
} hard, often requiring science education specialists to identify those
} alternative views and how to correct them.  The same evaluation needs
} to be done with standard practices, that is, after the issues needed
} to formulate them have been resolved.  And let's not even get started
} on the topic of theory-dependence!
}
} Best,
} Ellery
}
} ---------------
} Ellery Frahm
} Manager & Principal Analyst, Electron Microprobe Lab
} Senior Research Fellow, Geology and Geophysics
} Doctoral Candidate, Department of Anthropology
} University of Minnesota - Twin Cities
} http://web.mac.com/elleryfrahm/
}
} ==============================Original
Headers==============================
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} 5, 23 -- From: Ellery Frahm {frah0010-at-umn.edu}
} 5, 23 -- To: "protrain-at-emcourses.com" {protrain-at-emcourses.com} ,
} 5, 23 --         "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
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--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar


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From: protrain-at-emcourses.com
Date: Mon, 1 Mar 2010 09:30:34 -0600
Subject: [Microscopy] viaWWW: roughness measurments in the SEM

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

Hi Paul

I could have written your words!

It is my belief that the majority of operators simply run their SEM with
exactly the same parameters as the person who used it before them. There
are also too many labs where certain knobs (perhaps icons today) are taboo.

Even the more professional labs can make a hash of things. I will never
forget training a lady in cryo SEM who, several years later, took charge of
the SEM laboratory. I was back again training some new people a few weeks
into her new role, when she was hauled before her big boss. He complained
bitterly that since she took over the results had not been up to standard;
they looked nothing like what laboratory had been turning out under the
previous manager. Poor girl was in tears when she returned. When working
with polymer materials the kV is quite important so the earlier manager's
15kV specification had hardly fitted the bill; clearly her big boss had no
idea!

That was a very high level organisation with an international reputation for
science and technology; what hope do we have?

Just how many people teach an operator of a SEM to start at the lowest kV
and increase it step by step until the image information changes; that is
the point when the true surface image is disappearing!

And we worry about measuring surface roughness :-)

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: Gerroir, Paul [mailto:paul.gerroir-at-xrcc.xeroxlabs.com]
Sent: 01 March 2010 14:31
To: protrain-at-emcourses.com

Hi

I am always fascinated when people consider measuring surface roughness.
I
have had a mass of questions over the past few years on this subject due
to
the "developments" in anti wrinkle treatments; how rough is the surface?

Why I am fascinated is that as far as I am concerned the roughness of a
surface may be "modified" through the use of different imaging
techniques.
Crudely, to use a high kV and a surface looks rougher, use a low kV and
it
looks smooth, it all depends upon the surface/sub surface relationship
and
penetration.

An example - using rough emery paper grind flat the surface of an
aluminium
plate. Leave this for a day or two and then examine it in the SEM. At
low
kV it will probably charge and, even when coated, look smooth. At high
kV
you will penetrate through the oxide film and see the original rough
surface.

What is the general view on surface roughness monitoring, or is this so
gross in SEM terms that the instrument's interpretation of the image is
not
considered to be a problem.

I am interested in comment?


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide Tel +44
(0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com


-----Original Message-----
X-from: stevem-at-vt.edu [mailto:stevem-at-vt.edu]
Sent: 26 February 2010 01:12
To: protrain-at-emcourses.com

This Question/Comment was submitted to the Microscopy Listserver
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Email: stevem-at-vt.edu
Name: Steve McCartney

Organization: Va Tech

Title-Subject: [Filtered] roughness measurments in the SEM

Message: Hello: We are trying to get surface roughness measurements
on sample that has height variations from about 50nm to several
microns. We were not able to use AFM. I have read a little about
the software that can be used to measure roughness in the SEM. I am
wondering if this can be used to measure such extreme differences in
roughness and if so does anyone have any recommendations as to which
software to use. Also are there other techniques we should consider.
Thanks in advance for any help. Steve McCartney

Login Host: 198.82.148.215
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From: nantho2-at-emory.edu
Date: Mon, 1 Mar 2010 13:03:49 -0600
Subject: [Microscopy] Sample Tape

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

Hi all, has anybody had any experience with Secure Seal imaging
spacers? They essentially 'tape off' a small circular area on the slide
to hold the solution of interest, over which the cover slip is placed.
They appear to be very expense and I was wondering if anybody had any
ideas about inert adhesive tapes that might be available elsewhere. If
so, I wonder, is this tape available in different thicknesses? Maybe
0.1 or 0.05 mm.

Thanks for your time.

Neil


==============================Original Headers==============================
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From: larry.ackerman-at-ucsf.edu
Date: Mon, 1 Mar 2010 18:03:09 -0600
Subject: [Microscopy] N California freeze-fracture

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

I have a project that calls for freeze-fracture but have not found a
functioning unit in Northern California. Do you know of one?

--I do know about Brigitte Papahadjopoulos-Sternberg's NanoAnalytical
Laboratory but would prefer an academic, non-commercial lab.

Thanks,
Larry
--
Larry Ackerman, Specialist
Electron Microscopy Lab
UCSF, Dept. of Anatomy, Rm S1355
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

415-999-4758

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From: hanke-at-mee-inc.com
Date: Mon, 1 Mar 2010 18:25:25 -0600
Subject: [Microscopy] Re: roughness measurements in the SEM

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

Steve:

Good comments by all about the caveats to do this by SEM. As with any
measurement, especially surface roughness, it is important to understand
the limitations of your technique, the objectives of the evaluation, and
have a method to validate. Very basic roughness determinations using
contact profilometers have cut-off value inputs that would be somewhat
analogous to the accelerating voltage variables in SEM images. AFM has
limitations based on the tip geometry. Interferometry has issues with
reflectivity and light transmission. So take your pick of the potential
measurement anomalies and errors and choose the method that suits your
sample.

I have used the MeX software successfully in some cases; not so
successfully in others. I suspect that it is better now than the older
version that we have. If you understand what you are trying to
accomplish and your technique, you will have a shot at producing valid
results. Good luck.

--
Larry D. Hanke, P.E.
Materials Evaluation and Engineering, Inc.
Practical Solutions Through Technology and Innovation
http://www.mee-inc.com (763) 449-8870


} Message: Hello: We are trying to get surface roughness measurements
} on sample that has height variations from about 50nm to several
} microns. We were not able to use AFM. I have read a little about
} the software that can be used to measure roughness in the SEM. I am
} wondering if this can be used to measure such extreme differences in
} roughness and if so does anyone have any recommendations as to which
} software to use. Also are there other techniques we should consider.
} Thanks in advance for any help. Steve McCartney


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From: PhillipsT-at-missouri.edu
Date: Mon, 1 Mar 2010 20:28:53 -0600
Subject: [Microscopy] Sample Tape

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

I haven't used this brand. The answer to your question depends on your application. If you are just trying to conserve antibody in immunocytochemical staining protocols, there are other brands and approaches. I use a different brand for my immunocytochemistry protocols that require {20 ul per slide. We are able to remove and wash the coverslips - occasionally breaking one but not too often. I remember someone pointing out the use of pinstripe tape from an autoparts store one time for this purpose. I didn't try it but it sounded like a great idea.

Thomas E. Phillips, Ph.D.
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
Biological Sciences
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (voice)
573-882-0123 (fax)


-----Original Message-----
X-from: nantho2-at-emory.edu [mailto:nantho2-at-emory.edu]
Sent: Monday, March 01, 2010 1:05 PM
To: Phillips, Thomas E.

Hi all, has anybody had any experience with Secure Seal imaging
spacers? They essentially 'tape off' a small circular area on the slide
to hold the solution of interest, over which the cover slip is placed.
They appear to be very expense and I was wondering if anybody had any
ideas about inert adhesive tapes that might be available elsewhere. If
so, I wonder, is this tape available in different thicknesses? Maybe
0.1 or 0.05 mm.

Thanks for your time.

Neil


==============================Original Headers==============================
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From: nizets2-at-yahoo.com
Date: Tue, 2 Mar 2010 05:51:47 -0600
Subject: [Microscopy] Re: roughness measurements in the SEM

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

I can only agree with Larry. All techniques fit to all situations. Actually being able to use the right technique for the right purpose is one thing that makes a good scientist from a bad one, or better said an experienced scientist from a beginner.

As for the kV-in-SEM debate, perhaps one comment: you are talking about absolute values, but it may just be that the user wants to know the impact of a treatment on the roughness. In this case, as long as you use similar parameters for examination, it makes sense because you are working on a relative scale: how does the roughness changes relative to a control (untreated) specimen.

Stephane



----- Original Message ----
X-from: "hanke-at-mee-inc.com" {hanke-at-mee-inc.com}
To: nizets2-at-yahoo.com
Sent: Tue, March 2, 2010 1:28:09 AM

Steve:

Good comments by all about the caveats to do this by SEM. As with any
measurement, especially surface roughness, it is important to understand
the limitations of your technique, the objectives of the evaluation, and
have a method to validate. Very basic roughness determinations using
contact profilometers have cut-off value inputs that would be somewhat
analogous to the accelerating voltage variables in SEM images. AFM has
limitations based on the tip geometry. Interferometry has issues with
reflectivity and light transmission. So take your pick of the potential
measurement anomalies and errors and choose the method that suits your
sample.

I have used the MeX software successfully in some cases; not so
successfully in others. I suspect that it is better now than the older
version that we have. If you understand what you are trying to
accomplish and your technique, you will have a shot at producing valid
results. Good luck.

--
Larry D. Hanke, P.E.
Materials Evaluation and Engineering, Inc.
Practical Solutions Through Technology and Innovation
http://www.mee-inc.com          (763) 449-8870


} Message: Hello:  We are trying to get surface roughness measurements
} on sample that has height variations from about 50nm to several
} microns.  We were not able to use AFM.  I have read a little about
} the software that can be used to measure roughness in the SEM.  I am
} wondering if this can be used to measure such extreme differences in
} roughness and if so does anyone have any recommendations as to which
} software to use.  Also are there other techniques we should consider.
} Thanks in advance for any help.  Steve McCartney


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From: jacques.faerber-at-ipcms.u-strasbg.fr
Date: Tue, 2 Mar 2010 08:37:15 -0600
Subject: [Microscopy] Re: viaWWW: roughness measurments in the SEM

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

Hi all

Two cents about this subject(s) :

First cent :
} S. Chapman wrote : Just how many people teach an operator of a SEM to start at the lowest kV
} and increase it step by step until the image information changes; that is
} the point when the true surface image is disappearing!
Praticaly, in most case one uses the inverse way : one begin at a high
energy, say 15 keV, and one go bravely "down" to 10 keV. No difference ?
OK, a bit further... 5 keV ? There is somthing different but... One
begin to be reckless and one go down to 3 keV.....And so on. Only real
crazy people will go down to 0.5 keV !!! ( ;-) Maybe I'm crazy !)
Why is it so difficult to change this approach ? OK, there is first what
was soon mentionned as the "traditions", "lab cultures", "protocols",
which have a high weight. I see too two other reasons, a subjective and
a more objective.
Subjectively, a high energy picture looks more contrasted and seems
more pleasent to look at. One think that's better, sharper etc. One make
a confusion between contrast, sharpness and information, and information
at which scale.
On the other hand, more objectively, it's a practical reality then
as one change the energy, one have always to correcte more or less the
alignements, astigmatism and focus for each change, and one have/take
sometimes not the time or the motivation to do all this job for each
sample again and again. One search "correct" conditions for a type of
sample and one keep them for the followings until their inadaptation
comes out obviously. If there is some need to do EDS, one must go to
higher energies, higher current, and restart these settings again. One
think at the 10 or 15 samples which are waiting on the desk, and at the
clock which is running further, etc. I must confess that sometimes, at
5:30 PM, after a day of work, when the eyes are tired from the
semidarknes ( and the sun shines outside...), I'm too follish to try an
other set of conditions...

Second cent

To come back to the initial question, as we had some samples with too
steep slopes for AFM, I tried two sftwares for 3D reconstruction from
tilted SEM images (Mex by Alicona, soon mentionned, and 3DtopX by
SamX/ONERA).
Each uses a different mathematical approach to do the reconstruction,
but have in common to try to make a position correlation pixel by pixel
between the corresponding pixels of 2 or 3 images taken at different
tilt angles, to reconstruct the 3D structure of the field of view.
The main difficulty I've encountered is the way in which the noise of
the picture is taken in account. A greylevel variation is considered as
a variation in hight or orientation. So if there is some noise in the
image, there must be a sort of smoothing to deal with it. But where is
the limit between noise and smale rugosity ? How do you get unnoised
images at magnification and energy conditions to resolve the low level
rugosity of a surface, with tilt angles up to 30 to 40°, and the
corresponding needed WD ? From the 50 nm up to several microns scale,
you will have some difficulties to get pictures without noise and/or
with the smale scale rugosity information together with the large scale one.
An other limitation is the interpretation of uniform and very flat
surfaces, where there is no greylevel variations. The alorithm, will
not be able to locate the coresponding pixels, as their neighbourhood is
uniform. A light noise will be understood as a smale rugosity or reversely.
A more practical aspect of these tools is the need to take sets of well
choosen pictures, as much on the sample side, as on the needs for the
reconstruction procedure. It's time consuming, if one want some
statistical results. The user has less control how the parameters of the
algorithm are defined. You get nice 3D reconstructions, where you can
get nice profiles, with a lot of rugosity parameters (Ra, RDIN, Rrms,
etc), but you know less about their valadity for your study. It's to the
user to validate the methode. Probably there are some typical samples
and domaines where they are usefull, but there are several limitations too.

Jacques

J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr



stevem-at-vt.edu a écrit :
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} Title-Subject: [Filtered] roughness measurments in the SEM
}
} Message: Hello: We are trying to get surface roughness measurements
} on sample that has height variations from about 50nm to several
} microns. We were not able to use AFM. I have read a little about
} the software that can be used to measure roughness in the SEM. I am
} wondering if this can be used to measure such extreme differences in
} roughness and if so does anyone have any recommendations as to which
} software to use. Also are there other techniques we should consider.
} Thanks in advance for any help. Steve McCartney
}
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From: protrain-at-emcourses.com
Date: Tue, 2 Mar 2010 09:26:11 -0600
Subject: [Microscopy] Re: viaWWW: roughness measurments in the SEM

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

Hi Jacques

I am afraid I cannot agree with you on a number of counts

1. Starting from a high kV means that you may charge the specimen,
cause heat damage, etc, even before you have any idea what the specimen
looks like.

[I once helped out a professor who had grown cells on glass with unique
results; he was so excited. Sure, very few people look at grilled cells,
his work was unique thanks to a rubbish sputter coater which cooked them for
him! How did I find out? I put un unused preparation in the SEM at 2kV
without coating, when for about 30 seconds we could see the real structure
upon which we based our correction of the coating technique. Have to say
the day with this gut got me fired as I was only instructed to spend half an
hour with each customer. So I came back from America and started Protrain;
28 years ago.]

2. Why you like the higher kV image is due to the contribution made by
BSE and converted BSE (SE3) which indicates a degree of sub surface
information, which if we really want to know about the surface is not such a
good idea!

I do agree that higher kV images are often easier to obtain and they also
are often "prettier", this is due to the BSE contribution by which ever
means, but they never show the true surface!

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com


-----Original Message-----
X-from: jacques.faerber-at-ipcms.u-strasbg.fr
[mailto:jacques.faerber-at-ipcms.u-strasbg.fr]
Sent: 02 March 2010 14:38
To: protrain-at-emcourses.com

Hi all

Two cents about this subject(s) :

First cent :
} S. Chapman wrote : Just how many people teach an operator of a SEM to
start at the lowest kV
} and increase it step by step until the image information changes; that is
} the point when the true surface image is disappearing!
Praticaly, in most case one uses the inverse way : one begin at a high
energy, say 15 keV, and one go bravely "down" to 10 keV. No difference ?
OK, a bit further... 5 keV ? There is somthing different but... One
begin to be reckless and one go down to 3 keV.....And so on. Only real
crazy people will go down to 0.5 keV !!! ( ;-) Maybe I'm crazy !)
Why is it so difficult to change this approach ? OK, there is first what
was soon mentionned as the "traditions", "lab cultures", "protocols",
which have a high weight. I see too two other reasons, a subjective and
a more objective.
Subjectively, a high energy picture looks more contrasted and seems
more pleasent to look at. One think that's better, sharper etc. One make
a confusion between contrast, sharpness and information, and information
at which scale.
On the other hand, more objectively, it's a practical reality then
as one change the energy, one have always to correcte more or less the
alignements, astigmatism and focus for each change, and one have/take
sometimes not the time or the motivation to do all this job for each
sample again and again. One search "correct" conditions for a type of
sample and one keep them for the followings until their inadaptation
comes out obviously. If there is some need to do EDS, one must go to
higher energies, higher current, and restart these settings again. One
think at the 10 or 15 samples which are waiting on the desk, and at the
clock which is running further, etc. I must confess that sometimes, at
5:30 PM, after a day of work, when the eyes are tired from the
semidarknes ( and the sun shines outside...), I'm too follish to try an
other set of conditions...

Second cent

To come back to the initial question, as we had some samples with too
steep slopes for AFM, I tried two sftwares for 3D reconstruction from
tilted SEM images (Mex by Alicona, soon mentionned, and 3DtopX by
SamX/ONERA).
Each uses a different mathematical approach to do the reconstruction,
but have in common to try to make a position correlation pixel by pixel
between the corresponding pixels of 2 or 3 images taken at different
tilt angles, to reconstruct the 3D structure of the field of view.
The main difficulty I've encountered is the way in which the noise of
the picture is taken in account. A greylevel variation is considered as
a variation in hight or orientation. So if there is some noise in the
image, there must be a sort of smoothing to deal with it. But where is
the limit between noise and smale rugosity ? How do you get unnoised
images at magnification and energy conditions to resolve the low level
rugosity of a surface, with tilt angles up to 30 to 40°, and the
corresponding needed WD ? From the 50 nm up to several microns scale,
you will have some difficulties to get pictures without noise and/or
with the smale scale rugosity information together with the large scale one.
An other limitation is the interpretation of uniform and very flat
surfaces, where there is no greylevel variations. The alorithm, will
not be able to locate the coresponding pixels, as their neighbourhood is
uniform. A light noise will be understood as a smale rugosity or reversely.
A more practical aspect of these tools is the need to take sets of well
choosen pictures, as much on the sample side, as on the needs for the
reconstruction procedure. It's time consuming, if one want some
statistical results. The user has less control how the parameters of the
algorithm are defined. You get nice 3D reconstructions, where you can
get nice profiles, with a lot of rugosity parameters (Ra, RDIN, Rrms,
etc), but you know less about their valadity for your study. It's to the
user to validate the methode. Probably there are some typical samples
and domaines where they are usefull, but there are several limitations too.

Jacques

J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr



stevem-at-vt.edu a écrit :
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} Organization: Va Tech
}
} Title-Subject: [Filtered] roughness measurments in the SEM
}
} Message: Hello: We are trying to get surface roughness measurements
} on sample that has height variations from about 50nm to several
} microns. We were not able to use AFM. I have read a little about
} the software that can be used to measure roughness in the SEM. I am
} wondering if this can be used to measure such extreme differences in
} roughness and if so does anyone have any recommendations as to which
} software to use. Also are there other techniques we should consider.
} Thanks in advance for any help. Steve McCartney
}
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27, 28 -- From protrain-at-emcourses.com Tue Mar 2 09:26:11 2010
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From: jacques.faerber-at-ipcms.u-strasbg.fr
Date: Tue, 2 Mar 2010 10:33:39 -0600
Subject: [Microscopy] Re: viaWWW: roughness measurments in the SEM

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

OK Steve

My english is not so good, and I suppose I missexplaine my thought.

I fully agree with your answer, and with your former comment I cited. I
think we are at the same tune on that question ! By the way, a short
article in Europeen Microscopy, in the begining 1990 I believe, from...
S. Chapman ! changed my way of doing SEM. But the first SEM I used was
able to go up to 60 keV, and not lower then 10 keV...

In most cases, I begin at a low energy (2-3 keV) and from that point I
try lower or higher, but I dont begin "at the lowest possible". It's a
compromise, but I've not enough time to make a systematic exploration of
the possible settings on each sample. Possibly the conditions are not
always the best !
On the other hand, at less then 500 eV, there are often uncontrolled
and sometimes unexplanables contrasts (SE yield variations, giving some
compo effects, in particular with organics).

I wanted to point out in my former mail that the common way is more to
begin at high energy and lower it after, and that for the reasons I
explaned. I'm not justifying these reasons, I just observed them. But
it's always usefull to understand "why" do people work the wrong way.

Hoping it's clearer !

Jacques

J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr



protrain-at-emcourses.com a écrit :
}
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} Hi Jacques
}
} I am afraid I cannot agree with you on a number of counts
}
} 1. Starting from a high kV means that you may charge the specimen,
} cause heat damage, etc, even before you have any idea what the specimen
} looks like.
}
} [I once helped out a professor who had grown cells on glass with unique
} results; he was so excited. Sure, very few people look at grilled cells,
} his work was unique thanks to a rubbish sputter coater which cooked them for
} him! How did I find out? I put un unused preparation in the SEM at 2kV
} without coating, when for about 30 seconds we could see the real structure
} upon which we based our correction of the coating technique. Have to say
} the day with this gut got me fired as I was only instructed to spend half an
} hour with each customer. So I came back from America and started Protrain;
} 28 years ago.]
}
} 2. Why you like the higher kV image is due to the contribution made by
} BSE and converted BSE (SE3) which indicates a degree of sub surface
} information, which if we really want to know about the surface is not such a
} good idea!
}
} I do agree that higher kV images are often easier to obtain and they also
} are often "prettier", this is due to the BSE contribution by which ever
} means, but they never show the true surface!
}
} Steve
}
} Steve Chapman FRMS
} Senior Consultant Protrain
} For consultancy and training in electron microscopy world wide
} Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
} www.emcourses.com
}
}
} -----Original Message-----
} X-from: jacques.faerber-at-ipcms.u-strasbg.fr
} [mailto:jacques.faerber-at-ipcms.u-strasbg.fr]
} Sent: 02 March 2010 14:38
} To: protrain-at-emcourses.com
} Subject: [Microscopy] Re: viaWWW: roughness measurments in the SEM
}
}
}
}
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} Hi all
}
} Two cents about this subject(s) :
}
} First cent :
}
} } S. Chapman wrote : Just how many people teach an operator of a SEM to
} }
} start at the lowest kV
}
} } and increase it step by step until the image information changes; that is
} } the point when the true surface image is disappearing!
} }
} Praticaly, in most case one uses the inverse way : one begin at a high
} energy, say 15 keV, and one go bravely "down" to 10 keV. No difference ?
} OK, a bit further... 5 keV ? There is somthing different but... One
} begin to be reckless and one go down to 3 keV.....And so on. Only real
} crazy people will go down to 0.5 keV !!! ( ;-) Maybe I'm crazy !)
} Why is it so difficult to change this approach ? OK, there is first what
} was soon mentionned as the "traditions", "lab cultures", "protocols",
} which have a high weight. I see too two other reasons, a subjective and
} a more objective.
} Subjectively, a high energy picture looks more contrasted and seems
} more pleasent to look at. One think that's better, sharper etc. One make
} a confusion between contrast, sharpness and information, and information
} at which scale.
} On the other hand, more objectively, it's a practical reality then
} as one change the energy, one have always to correcte more or less the
} alignements, astigmatism and focus for each change, and one have/take
} sometimes not the time or the motivation to do all this job for each
} sample again and again. One search "correct" conditions for a type of
} sample and one keep them for the followings until their inadaptation
} comes out obviously. If there is some need to do EDS, one must go to
} higher energies, higher current, and restart these settings again. One
} think at the 10 or 15 samples which are waiting on the desk, and at the
} clock which is running further, etc. I must confess that sometimes, at
} 5:30 PM, after a day of work, when the eyes are tired from the
} semidarknes ( and the sun shines outside...), I'm too follish to try an
} other set of conditions...
}
} Second cent
}
} To come back to the initial question, as we had some samples with too
} steep slopes for AFM, I tried two sftwares for 3D reconstruction from
} tilted SEM images (Mex by Alicona, soon mentionned, and 3DtopX by
} SamX/ONERA).
} Each uses a different mathematical approach to do the reconstruction,
} but have in common to try to make a position correlation pixel by pixel
} between the corresponding pixels of 2 or 3 images taken at different
} tilt angles, to reconstruct the 3D structure of the field of view.
} The main difficulty I've encountered is the way in which the noise of
} the picture is taken in account. A greylevel variation is considered as
} a variation in hight or orientation. So if there is some noise in the
} image, there must be a sort of smoothing to deal with it. But where is
} the limit between noise and smale rugosity ? How do you get unnoised
} images at magnification and energy conditions to resolve the low level
} rugosity of a surface, with tilt angles up to 30 to 40°, and the
} corresponding needed WD ? From the 50 nm up to several microns scale,
} you will have some difficulties to get pictures without noise and/or
} with the smale scale rugosity information together with the large scale one.
} An other limitation is the interpretation of uniform and very flat
} surfaces, where there is no greylevel variations. The alorithm, will
} not be able to locate the coresponding pixels, as their neighbourhood is
} uniform. A light noise will be understood as a smale rugosity or reversely.
} A more practical aspect of these tools is the need to take sets of well
} choosen pictures, as much on the sample side, as on the needs for the
} reconstruction procedure. It's time consuming, if one want some
} statistical results. The user has less control how the parameters of the
} algorithm are defined. You get nice 3D reconstructions, where you can
} get nice profiles, with a lot of rugosity parameters (Ra, RDIN, Rrms,
} etc), but you know less about their valadity for your study. It's to the
} user to validate the methode. Probably there are some typical samples
} and domaines where they are usefull, but there are several limitations too.
}
} Jacques
}
} J. Faerber
} IPCMS-GSI
} (Institut de Physique et Chimie des Matériaux de Strasbourg
} Groupe Surface et Interfaces)
} 23, rue de Loess ; BP43
} 67034 Strasbourg CEDEX 2
} France
}
} Tel 00 33(0)3 88 10 71 01
} Fax 00 33(0)3 88 10 72 48
} E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr
}
}
}
} stevem-at-vt.edu a écrit :
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} } Email: stevem-at-vt.edu
} } Name: Steve McCartney
} }
} } Organization: Va Tech
} }
} } Title-Subject: [Filtered] roughness measurments in the SEM
} }
} } Message: Hello: We are trying to get surface roughness measurements
} } on sample that has height variations from about 50nm to several
} } microns. We were not able to use AFM. I have read a little about
} } the software that can be used to measure roughness in the SEM. I am
} } wondering if this can be used to measure such extreme differences in
} } roughness and if so does anyone have any recommendations as to which
} } software to use. Also are there other techniques we should consider.
} } Thanks in advance for any help. Steve McCartney
} }
} } Login Host: 198.82.148.215
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From: peter.eschbach-at-hp.com
Date: Tue, 2 Mar 2010 11:35:22 -0600
Subject: [Microscopy] Re: viaWWW: roughness measurments in the SEM

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



Steven:

OK, I think I have the correct listserver address and you as the originator of this thread now - I learn a little slowly these days :o)

The notes from Chapman and others were great to read, but I am wondering if anybody has suggested Scanning White Light Interferometry (SWLI) to you? I consulted with coworkers at HP and they think 50 nm to 2 microns is right in the "wheelhouse" or capability of SWLI. A couple of tricks or "flys in the ointment" : if the 50 nm features are very sharply peaked, then you will only see the tops of the features, and make sure to run it in PSI (phase sensitive interferometry - I hope I got that right as I am NOT a SWLI guy :o) ). Oh yes, and to get a good SWLI image, the surface must be reflective - so coating non reflective surfaces with thin metal is a key.

Pete Eschbach
TEM/Nanonidneter Engineer
Hewlett Packard Corp
1000 NE Circle Blvd.
Corvallis, OR 97330-4241
541 715 1701

-----Original Message-----
X-from: jacques.faerber-at-ipcms.u-strasbg.fr [mailto:jacques.faerber-at-ipcms.u-strasbg.fr]
Sent: Tuesday, March 02, 2010 8:44 AM
To: Eschbach, Peter

OK Steve

My english is not so good, and I suppose I missexplaine my thought.

I fully agree with your answer, and with your former comment I cited. I think we are at the same tune on that question ! By the way, a short article in Europeen Microscopy, in the begining 1990 I believe, from...
S. Chapman ! changed my way of doing SEM. But the first SEM I used was able to go up to 60 keV, and not lower then 10 keV...

In most cases, I begin at a low energy (2-3 keV) and from that point I try lower or higher, but I dont begin "at the lowest possible". It's a compromise, but I've not enough time to make a systematic exploration of the possible settings on each sample. Possibly the conditions are not always the best !
On the other hand, at less then 500 eV, there are often uncontrolled and sometimes unexplanables contrasts (SE yield variations, giving some compo effects, in particular with organics).

I wanted to point out in my former mail that the common way is more to begin at high energy and lower it after, and that for the reasons I explaned. I'm not justifying these reasons, I just observed them. But it's always usefull to understand "why" do people work the wrong way.

Hoping it's clearer !

Jacques

J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr



protrain-at-emcourses.com a écrit :
}
} ----------------------------------------------------------------------
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}
} Hi Jacques
}
} I am afraid I cannot agree with you on a number of counts
}
} 1. Starting from a high kV means that you may charge the specimen,
} cause heat damage, etc, even before you have any idea what the
} specimen looks like.
}
} [I once helped out a professor who had grown cells on glass with
} unique results; he was so excited. Sure, very few people look at
} grilled cells, his work was unique thanks to a rubbish sputter coater
} which cooked them for him! How did I find out? I put un unused
} preparation in the SEM at 2kV without coating, when for about 30
} seconds we could see the real structure upon which we based our
} correction of the coating technique. Have to say the day with this
} gut got me fired as I was only instructed to spend half an hour with
} each customer. So I came back from America and started Protrain;
} 28 years ago.]
}
} 2. Why you like the higher kV image is due to the contribution made by
} BSE and converted BSE (SE3) which indicates a degree of sub surface
} information, which if we really want to know about the surface is not
} such a good idea!
}
} I do agree that higher kV images are often easier to obtain and they
} also are often "prettier", this is due to the BSE contribution by
} which ever means, but they never show the true surface!
}
} Steve
}
} Steve Chapman FRMS
} Senior Consultant Protrain
} For consultancy and training in electron microscopy world wide Tel +44
} (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
} www.emcourses.com
}
}
} -----Original Message-----
} X-from: jacques.faerber-at-ipcms.u-strasbg.fr
} [mailto:jacques.faerber-at-ipcms.u-strasbg.fr]
} Sent: 02 March 2010 14:38
} To: protrain-at-emcourses.com
} Subject: [Microscopy] Re: viaWWW: roughness measurments in the SEM
}
}
}
}
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} Hi all
}
} Two cents about this subject(s) :
}
} First cent :
}
} } S. Chapman wrote : Just how many people teach an operator of a SEM to
} }
} start at the lowest kV
}
} } and increase it step by step until the image information changes;
} } that is the point when the true surface image is disappearing!
} }
} Praticaly, in most case one uses the inverse way : one begin at a high
} energy, say 15 keV, and one go bravely "down" to 10 keV. No difference ?
} OK, a bit further... 5 keV ? There is somthing different but... One
} begin to be reckless and one go down to 3 keV.....And so on. Only real
} crazy people will go down to 0.5 keV !!! ( ;-) Maybe I'm crazy !) Why
} is it so difficult to change this approach ? OK, there is first what
} was soon mentionned as the "traditions", "lab cultures", "protocols",
} which have a high weight. I see too two other reasons, a subjective
} and a more objective.
} Subjectively, a high energy picture looks more contrasted and
} seems more pleasent to look at. One think that's better, sharper etc.
} One make a confusion between contrast, sharpness and information, and
} information at which scale.
} On the other hand, more objectively, it's a practical reality then
} as one change the energy, one have always to correcte more or less the
} alignements, astigmatism and focus for each change, and one have/take
} sometimes not the time or the motivation to do all this job for each
} sample again and again. One search "correct" conditions for a type of
} sample and one keep them for the followings until their inadaptation
} comes out obviously. If there is some need to do EDS, one must go to
} higher energies, higher current, and restart these settings again. One
} think at the 10 or 15 samples which are waiting on the desk, and at
} the clock which is running further, etc. I must confess that
} sometimes, at 5:30 PM, after a day of work, when the eyes are tired
} from the semidarknes ( and the sun shines outside...), I'm too follish
} to try an other set of conditions...
}
} Second cent
}
} To come back to the initial question, as we had some samples with too
} steep slopes for AFM, I tried two sftwares for 3D reconstruction from
} tilted SEM images (Mex by Alicona, soon mentionned, and 3DtopX by
} SamX/ONERA).
} Each uses a different mathematical approach to do the reconstruction,
} but have in common to try to make a position correlation pixel by
} pixel between the corresponding pixels of 2 or 3 images taken at
} different tilt angles, to reconstruct the 3D structure of the field of view.
} The main difficulty I've encountered is the way in which the noise of
} the picture is taken in account. A greylevel variation is considered
} as a variation in hight or orientation. So if there is some noise in
} the image, there must be a sort of smoothing to deal with it. But
} where is the limit between noise and smale rugosity ? How do you get
} unnoised images at magnification and energy conditions to resolve the
} low level rugosity of a surface, with tilt angles up to 30 to 40°, and
} the corresponding needed WD ? From the 50 nm up to several microns
} scale, you will have some difficulties to get pictures without noise
} and/or with the smale scale rugosity information together with the large scale one.
} An other limitation is the interpretation of uniform and very flat
} surfaces, where there is no greylevel variations. The alorithm, will
} not be able to locate the coresponding pixels, as their neighbourhood
} is uniform. A light noise will be understood as a smale rugosity or reversely.
} A more practical aspect of these tools is the need to take sets of
} well choosen pictures, as much on the sample side, as on the needs for
} the reconstruction procedure. It's time consuming, if one want some
} statistical results. The user has less control how the parameters of
} the algorithm are defined. You get nice 3D reconstructions, where you
} can get nice profiles, with a lot of rugosity parameters (Ra, RDIN,
} Rrms, etc), but you know less about their valadity for your study.
} It's to the user to validate the methode. Probably there are some
} typical samples and domaines where they are usefull, but there are several limitations too.
}
} Jacques
}
} J. Faerber
} IPCMS-GSI
} (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe
} Surface et Interfaces) 23, rue de Loess ; BP43
} 67034 Strasbourg CEDEX 2
} France
}
} Tel 00 33(0)3 88 10 71 01
} Fax 00 33(0)3 88 10 72 48
} E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr
}
}
}
} stevem-at-vt.edu a écrit :
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}
} } Email: stevem-at-vt.edu
} } Name: Steve McCartney
} }
} } Organization: Va Tech
} }
} } Title-Subject: [Filtered] roughness measurments in the SEM
} }
} } Message: Hello: We are trying to get surface roughness measurements
} } on sample that has height variations from about 50nm to several
} } microns. We were not able to use AFM. I have read a little about
} } the software that can be used to measure roughness in the SEM. I am
} } wondering if this can be used to measure such extreme differences in
} } roughness and if so does anyone have any recommendations as to which
} } software to use. Also are there other techniques we should consider.
} } Thanks in advance for any help. Steve McCartney
} }
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From: takenomm-at-u.washington.edu
Date: Tue, 2 Mar 2010 17:56:08 -0600
Subject: [Microscopy] Re: Image Colorizing

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

Hi

Yes it is much more clear than me writing in French - well done!

I once gave a lecture for a French pathology society meeting, but in
English, 400 people were in the lecture hall until I said I would have to
talk in English ~200 walked out! Some would say that is typical of one of
my lectures ;-)

Steve

-----Original Message-----
X-from: j.faerber [mailto:jacques.faerber-at-ipcms.u-strasbg.fr]
Sent: 02 March 2010 16:33
To: protrain-at-emcourses.com; Microscopy-at-microscopy.com

In addition to GIMP, there is a modification called GIMPshop that
attempts to make GIMP look and feel more like Photoshop.

http://en.wikipedia.org/wiki/GIMPshop

Most notably, GIMPshop allows the use of Photoshop plugins, which can be
useful.

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

Thank you all for the info. It's clear that I'd be best off with
Photoshop but since I don't have that option at the moment I'l try
GIMP and see how that goes.

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--
Marc Takeno, Ph.D. | Research Scientist | University of Washington
Department of Bioengineering | N330B Foege | Box 355061
(206) 543-4290 | takenomm-at-u.washington.edu


==============================Original Headers==============================
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From: r-holdford-at-ti.com
Date: Tue, 2 Mar 2010 21:08:48 -0600
Subject: [Microscopy] Re: roughness measurements in the SEM

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

Steve: as far as protocols go, I'm usually the one setting them for my
group. We sometimes look at polymers that coat the surface of
integrated circuits to see the differences different treatments have on
the surface finish. I don't do absolute measurements with the SEM but
comparisons to known good surfaces. I set the SEM conditions for others
to use, as our overseas assembly/test sites do not have AFM capability.

I would agree that "average" SEM users (those with not much experience
or those with not much desire to experiment) wouldn't think to use the
kV that will give the best information, not the nicest picture. It
comes down to training and willingness to learn. One lab I worked in,
the "average" user had 10+ years of SEM experience. The lab I'm in now,
some users would be doing well to get an image at any kV.

When I am training others on the SEM, I teach them to try different kVs
to see what needs seeing. Depth of penetration is a thing we discuss at
length, as it relates to images and EDX analysis. If you are having
bonding problems, use 1kV to see if there is any film on the surface of
the bonding pad or solder ball. If you need to see the metal layers
under the surface oxide, use around 20kV.


On 2/27/10 3:18 PM, protrain-at-emcourses.com wrote:
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
}
} Hi Paul
}
} That is exactly my point, change the kV change the information. Is there a
} specification for investigating surface roughness that states that the
} scientist should work at low kV; how low is low?
}
} My experience is that people will work at the kV that demonstrates the most
} attractive image of their material, is that the image that they will use to
} examine roughness? Often the most attractive image contains a degree of sub
} surface information which adds contrast, could that be used for surface
} roughness investigations?
}
} I work around the world with different cultures, different instruments and
} many different materials. I am not convinced that the average SEM operator
} would work at a kV that will only show the true specimen surface, it's not
} in their nature; you are talking of {2kV, or even lower for some materials?
}
} Steve
}
} -----Original Message-----
} X-from: Gerroir, Paul [mailto:paul.gerroir-at-xrcc.xeroxlabs.com]
} Sent: 26 February 2010 20:19
} To: protrain-at-emcourses.com
} Subject: RE: [Microscopy] Re: roughness measurements in the SEM
}
} Steve,
} Your comments on high/low kV and surface roughness might give people the
} wrong impression. Low kV allows better imaging of fine surface detail
} while higher kV penetrates more deeply and hence does not provide an
} image of the 'true' surface. From my experience this is certainly the
} case for polymers.
}
} Paul
}
} Paul J. Gerroir
}
} Microscopy
} Materials Characterization
} Xerox Research Centre of Canada
} 2660 Speakman Drive
} Mississauga, Ontario L5K 2L1
}
} Phone: 905-823-7091, ext.216
} FAX: 905-822-7022
} e-mail: paul.gerroir-at-xerox.com
}
}
}
} -----Original Message-----
} X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
} Sent: Friday, February 26, 2010 11:16 AM
} To: Gerroir, Paul
} Subject: [Microscopy] Re: roughness measurements in the SEM
}
}
}
}
} ------------------------------------------------------------------------
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} Hi
}
} I am always fascinated when people consider measuring surface roughness.
} I
} have had a mass of questions over the past few years on this subject due
} to
} the "developments" in anti wrinkle treatments; how rough is the surface?
}
} Why I am fascinated is that as far as I am concerned the roughness of a
} surface may be "modified" through the use of different imaging
} techniques.
} Crudely, to use a high kV and a surface looks rougher, use a low kV and
} it
} looks smooth, it all depends upon the surface/sub surface relationship
} and
} penetration.
}
} An example - using rough emery paper grind flat the surface of an
} aluminium
} plate. Leave this for a day or two and then examine it in the SEM. At
} low
} kV it will probably charge and, even when coated, look smooth. At high
} kV
} you will penetrate through the oxide film and see the original rough
} surface.
}
} What is the general view on surface roughness monitoring, or is this so
} gross in SEM terms that the instrument's interpretation of the image is
} not
} considered to be a problem.
}
} I am interested in comment?
}
}
} Steve Chapman FRMS
} Senior Consultant Protrain
} For consultancy and training in electron microscopy world wide Tel +44
} (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
} www.emcourses.com
}
}
} -----Original Message-----
} X-from: stevem-at-vt.edu [mailto:stevem-at-vt.edu]
} Sent: 26 February 2010 01:12
} To: protrain-at-emcourses.com
} Subject: [Microscopy] viaWWW: roughness measurments in the SEM
}
}
}
}
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} Email: stevem-at-vt.edu
} Name: Steve McCartney
}
} Organization: Va Tech
}
} Title-Subject: [Filtered] roughness measurments in the SEM
}
} Message: Hello: We are trying to get surface roughness measurements
} on sample that has height variations from about 50nm to several
} microns. We were not able to use AFM. I have read a little about
} the software that can be used to measure roughness in the SEM. I am
} wondering if this can be used to measure such extreme differences in
} roughness and if so does anyone have any recommendations as to which
} software to use. Also are there other techniques we should consider.
} Thanks in advance for any help. Steve McCartney
}
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--
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Becky Holdford (r-holdford-at-ti.com)
972-995-2360
972-648-8743 (pager)
SC Packaging FA
Texas Instruments, Inc.
13536 N. Central Expressway MS 940
Dallas, TX 75243
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~


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From: mcgeejj-at-gmail.com
Date: Tue, 2 Mar 2010 22:23:25 -0600
Subject: [Microscopy] Job Opening: Focused Ion Beam (FIB) Microscopist

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

Knolls Atomic Power Laboratory (KAPL) in the Albany, NY area has an
opening for an experienced Microscopist specializing in Focused Ion
Beam (FIB) based materials characterization, testing, and development.
This is a newly created, full-time position; U.S. Citizenship
required.  The microscopist will be responsible for the new Dual-Beam
FIB laboratory and materials characterization and testing utilizing
this instrument.  The FIB lab is part of a material characterization
group that also includes TEM, Auger, ESCA, SEM, XRD, and EPMA
instrumentation.  The position description can be accessed at this
link:

http://jobview.monster.com/FIB-Microscopist-Job-Niskayuna-NY-US-86538490.aspx

US citizenship is required.  Feel free to contact me for any
additional info or questions.

Thanks for your interest.

Jim McGee
Lead Engineer, Chemistry Programs
BMPC-KAPL
Tel: 518-395-4612


==============================Original Headers==============================
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6, 32 -- Subject: Job Opening: Focused Ion Beam (FIB) Microscopist
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From: pete.finger-at-jax.org
Date: Wed, 3 Mar 2010 08:27:52 -0600
Subject: [Microscopy] viaWWW: OTOTO Artifact

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Email: pete.finger-at-jax.org
Name: Pete Finger

Organization: The Jackson Laboratory

Title-Subject: [Filtered] OTOTO Artifact

Message: In the past, I have been processing mouse inner ears using
the OTOTO method for a user with great success. The last couple of
runs have produced a strange artifact, specifically, the stereo cilia
of the hair cells look like they've been heavily coated with some
substance akin to what I can only describe as "beer batter".

I always use fresh, filtered TCH, have dehydrated with ethanol and
dried my samples with HMDS.

Has anyone else experienced a similar artifact when using the OTOTO method??

Any help/suggestions with this matter would be greatly appreciated.

Thanks much,

Pete

Login Host: 209.222.197.44
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From: Frank_Karl-at-lincolnelectric.com
Date: Wed, 3 Mar 2010 13:05:28 -0600
Subject: [Microscopy] line scan for Si

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


I’m attempting to determine if there is a change in Si concentration
between metal grain boundaries in a weld. I’ve polished the surface to 1
um diamond and etched the surface very lightly with 4% nitric acid in
methanol so I can find the boundaries. It’s been suggest I try a line scan
with my EDS.

I’m running at 4KEV which should be enough overvoltage for Si line at
1.73ev. I selected 4KEV to limit penetration into the sample and to keep
from overwhelming my detector with un-wanted X-rays.%

So far, I’m underwhelmed with the results. Si isn’t in very high
concentrations in the metal but we believe Si concentrates in the grain
boundaries.

Our “tribal†knowledge suggest a small amp constant and large dwell time.
Making the spot size seems counter productive. More signal, but less
precision on determining when you are in the grain boundary as compared to
being on the metal grain.

Of course, the change in Si level may simply be too small to detect.

Any thoughts, suggestion or comments would be welcome

Thank!!

Frank!!
--
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communication in error, please notify us immediately by
replying to the message and deleting it from your computer.
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The Lincoln Electric Company
**************************************************************


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From: protrain-at-emcourses.com
Date: Wed, 3 Mar 2010 13:31:22 -0600
Subject: [Microscopy] line scan for Si

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

Hi Frank

Is it a good idea to etch the boundary as this will create surface roughness that will make a line scan or BSE definition rather messy? Could you find the boundary in BSE to simplify the edge definition to exclude etching?

I have worked with some clients interested in grain boundaries and in spite of their reticence the unetched were by far the most informative.

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: Frank_Karl-at-lincolnelectric.com [mailto:Frank_Karl-at-lincolnelectric.com]
Sent: 03 March 2010 19:07
To: protrain-at-emcourses.com


I’m attempting to determine if there is a change in Si concentration
between metal grain boundaries in a weld. I’ve polished the surface to 1
um diamond and etched the surface very lightly with 4% nitric acid in
methanol so I can find the boundaries. It’s been suggest I try a line scan
with my EDS.

I’m running at 4KEV which should be enough overvoltage for Si line at
1.73ev. I selected 4KEV to limit penetration into the sample and to keep
from overwhelming my detector with un-wanted X-rays.%

So far, I’m underwhelmed with the results. Si isn’t in very high
concentrations in the metal but we believe Si concentrates in the grain
boundaries.

Our “tribal†knowledge suggest a small amp constant and large dwell time.
Making the spot size seems counter productive. More signal, but less
precision on determining when you are in the grain boundary as compared to
being on the metal grain.

Of course, the change in Si level may simply be too small to detect.

Any thoughts, suggestion or comments would be welcome

Thank!!

Frank!!
--
*************************************************************
Note:
The information contained in this message may be
privileged and confidential and protected from disclosure. If
the reader of this message is not the intended recipient, or
an employee or agent responsible for delivering this message
to the intended recipient, you are hereby notified that any
dissemination, distribution or copying of this communication
is strictly prohibited. If you have received this
communication in error, please notify us immediately by
replying to the message and deleting it from your computer.
Thank you,
The Lincoln Electric Company
**************************************************************


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==============================Original Headers==============================
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From: PhillipsT-at-missouri.edu
Date: Wed, 3 Mar 2010 13:50:31 -0600
Subject: [Microscopy] automated light microscope slide scanner

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

I have been asked by a PI on our campus to consider adding an automated microscope slide scanner (e.g., Aperio, ImageExpress, NanoZoomer, etc) to our campus core facility. I am unsure of the market demand. Does anyone who man=
ages a LM imaging core care to comment on this approach? Or someone who uses one routinely? I am also considering using an automated stage driven by metamorph to do this. I would just like to hear from others about how they approach this. Tom



Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/




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From: mcauliff-at-umdnj.edu
Date: Wed, 3 Mar 2010 14:12:35 -0600
Subject: [Microscopy] Re: Re: Image Colorizing

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

Corel Paint Shop Pro Photo is also a very powerful program that will do
(just about?) everything Photoshop will do for about $100 US. I even got
a 2gig USB drive with my copy.

Geoff

takenomm-at-u.washington.edu wrote:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} In addition to GIMP, there is a modification called GIMPshop that
} attempts to make GIMP look and feel more like Photoshop.
}
} http://en.wikipedia.org/wiki/GIMPshop
}
} Most notably, GIMPshop allows the use of Photoshop plugins, which can be
} useful.
}
} -------- Original Message --------
}
} Thank you all for the info. It's clear that I'd be best off with
} Photoshop but since I don't have that option at the moment I'l try
} GIMP and see how that goes.
}
} ==============================Original Headers==============================
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--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff-at-umdnj.edu
**********************************************



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From: bill.mcmanus-at-usu.edu
Date: Wed, 3 Mar 2010 20:47:02 -0600
Subject: [Microscopy] viaWWW: epi-fluorescent parts for Ziess Photomicroscope?

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

I suppose someone has done the study. Maybe someone can reference a paper or two that carries more authority than I can now offer.

I get concerned any time someone gives me an etched sample to analyze. Certainly the etching introduces some topographic contrast. That could lead to some geometric effects and uncertainties that could alter the x-ray results. However, my bigger concern is the nature of the etching in the first place. It works because of preferential attack. It may attack grain boundaries because of the slight difference in energy at the boundaries, or it may attack one phase faster than another due to its composition.

However, if etching attacks a phase because of its composition, does that not alter the composition of the remaining material?

It's about that time that I tell people that I can give them an answer, but it's going to be wrong. The question is only will it still be right enough to do them some good.

Frank, your case may be made all the tougher by using a low voltage. It may be that the etching effect is subtle, but you "feel" the effect of it more since your beam is interacting nearer to the surface.

hth,
Warren

-----Original Message-----
X-from: Frank_Karl-at-lincolnelectric.com [mailto:Frank_Karl-at-lincolnelectric.com]
Sent: Wednesday, March 03, 2010 1:06 PM
To: wesaia-at-iastate.edu

Sounds like you need a TEM......

John Mardinly

Western Digital

-----Original Message-----
X-from: wesaia-at-iastate.edu [mailto:wesaia-at-iastate.edu]
Sent: Wednesday, March 03, 2010 1:00 PM
To: John Mardinly

I suppose someone has done the study. Maybe someone can reference a paper or
two that carries more authority than I can now offer.

I get concerned any time someone gives me an etched sample to analyze.
Certainly the etching introduces some topographic contrast. That could lead
to some geometric effects and uncertainties that could alter the x-ray
results. However, my bigger concern is the nature of the etching in the
first place. It works because of preferential attack. It may attack grain
boundaries because of the slight difference in energy at the boundaries, or
it may attack one phase faster than another due to its composition.

However, if etching attacks a phase because of its composition, does that
not alter the composition of the remaining material?

It's about that time that I tell people that I can give them an answer, but
it's going to be wrong. The question is only will it still be right enough
to do them some good.

Frank, your case may be made all the tougher by using a low voltage. It may
be that the etching effect is subtle, but you "feel" the effect of it more
since your beam is interacting nearer to the surface.

hth,
Warren

-----Original Message-----
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[mailto:Frank_Karl-at-lincolnelectric.com]
Sent: Wednesday, March 03, 2010 1:06 PM
To: wesaia-at-iastate.edu

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Email: bill.mcmanus-at-usu.edu
Name: Bill McManus

Organization: Western Dairy Center, Utah State University

Title-Subject: [Filtered] epi-fluorescent parts for Ziess Photomicroscope?

Message: I am trying to resurrect an on Ziess Photomicroscope to do
some epi-fluorescent work. Does any one have a line on any parts?

Bill

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From: heytimj-at-gmail.com
Date: Wed, 3 Mar 2010 20:47:26 -0600
Subject: [Microscopy] viaWWW: Spurr's resin replacement

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Email: heytimj-at-gmail.com
Name: Tim

Organization: University of Florida

Title-Subject: [Filtered] Spurr's resin replacement

Message: I am looking for a suitable replacement for Spurr's resin
that is currently available for purchase. I am doing TEM work with
orchid seeds. Any suggestions? There seem to be so many different
epoxy resins out there.

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From: Brendan.J.Foran-at-aero.org
Date: Wed, 3 Mar 2010 20:48:01 -0600
Subject: [Microscopy] viaWWW: opening for a Member of the Technical Staff in the

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Email: Brendan.J.Foran-at-aero.org
Name: Brendan J Foran

Organization: The Aerospace Corporation

Title-Subject: [Filtered] opening for a Member of
the Technical Staff in the Microelectronics
Technology Department

Message: The Aerospace Corporation
( {http://www.aero.org} www.aero.org) in El
Segundo, CA has an opening for a Member of the
Technical Staff in the Microelectronics
Technology Department.

JOB DUTIES: ÝConduct fundamental and applied
research at the nanoscale on advanced materials
and structures for space systems applications.
ÝProvide analytical technical support to various
space system programs. ÝDevelop innovative
research programs using state of the art micro-
and nano-analytical equipment available in our
labs and at national user facilities and via
collaborative efforts with academic groups.
ÝPresent and publish results at scientific
meetings and in refereed journals. Ý

QUALIFICATIONS: ÝPh.D. in physics, materials
science, physical chemistry, electrical
engineering, or related technical discipline
required. ÝExpertise required in one or more of
the following analytical techniques: transmission
electron microscopy, near field optical or
scanning probe microscopy, or time-of-flight
secondary ion mass spectrometry. ÝPreference is
given for expertise related to the
characterization of process development,
reliability, or failure of microelectronic and/or
optoelectronic devices. ÝAdditional background or
expertise in mathematical analysis, device
physics, modeling and computer-based simulation
is desirable. ÝGood oral communication and
technical writing skills, and the ability to work
effectively as part of a team are also required.

Candidate must be able to obtain US Government security clearances.

Interested candidates must apply through
{http://www.aero.org/careers/jobs/index.html} http://www.aero.org/careers/jobs/index.html
and reference Requisition # 938.

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The Aerospace Corporation
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From: jflaci-at-ms.sapientia.ro
Date: Thu, 4 Mar 2010 01:00:02 -0600
Subject: [Microscopy] Teeth again... and sample holder question...

Contents Retrieved from Microscopy Listserver Archives
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Hello all!

After the help You kindly provided for
selecting the proper coating for teeth, i do have another problem.

1. One of my stomatologysts (former classmate in highschool) would like
to take micrographs from tooth surface, than treat that surface than
again micrographs an treat and so on, for 7-8 times.

My problem is, that after 2 or 3 rounds of vacuuming in the SEM, the
tooth will crack, and the whole thing is compromised, as the
treatment material gets into the cracks and alters the result.

Can any of You give me some advise how to avoid the cracks?

2. Does anybody have a 10mm WD sample holder for Jeol JSM-5200?
If possible, I would like to kindly ask You to take some photographs
so I could build my own. We have a pretty good machine shop at the
University, they can handle this kind of work.

Best regards,
Laci

PS:Of course coating is already not an option, but thanks anyway for the
help.
I will do the imaging at low kV(1-5) and small wd(10mm the minimum of the
instrument).


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From: dsoren-at-umich.edu
Date: Thu, 4 Mar 2010 07:30:07 -0600
Subject: [Microscopy] TEM liver

Contents Retrieved from Microscopy Listserver Archives
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Hello all,

Does anyone have a tried and true protocol for processing liver for
TEM? We seem to be having problems with poor fixation. We have been
perfusing with 3% p.f. and 2.5 % glutaraldehyde in Sorensen's buffer,
followed by immersion fixation in the same mixture. We post fix with
1 % osmium in the same buffer, dehydrate with a graded series of
ethanol, change to propylene oxide, and then infiltrate and embed in
Epon. The cells immediately adjacent to the blood vessels and right
on the cut edge of the tissue are well fixed, but the cells located
on the interior of the tissue exhibit little white holes. The tissue
had been cut to about 1x2x2 mm. Another problem that we have
encountered in the past is cleared areas that remind me of canals
running through the cytoplasm. Is this to be expected? We would
love to hear from advice from some liver experts out there.

Thanks,

Dotty


Dorothy Sorenson
Microscopy and Image-analysis Laboratory
Department of Cell and Developmental Biology
University Of Michigan Medical School
A807 BSRB
109 Zina Pitcher Place
Ann Arbor, MI 48109-2200
(734)763-1170
FAX (734)763-1166



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From: mcauliff-at-umdnj.edu
Date: Thu, 4 Mar 2010 08:43:29 -0600
Subject: [Microscopy] Re: automated light microscope slide scanner

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We had the slide set (~180 slides) we use for teaching histology to
medical students scanned by Aperio rather than buy a scanner.
I suppose it all depends on how much demand you have for the service.

Geoff

PhillipsT-at-missouri.edu wrote:
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} I have been asked by a PI on our campus to consider adding an automated microscope slide scanner (e.g., Aperio, ImageExpress, NanoZoomer, etc) to our campus core facility. I am unsure of the market demand. Does anyone who man=
} ages a LM imaging core care to comment on this approach? Or someone who uses one routinely? I am also considering using an automated stage driven by metamorph to do this. I would just like to hear from others about how they approach this. Tom
}
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
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} University of Missouri
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--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff-at-umdnj.edu
**********************************************



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From: jsiegmund-at-7thwavelabs.com
Date: Thu, 4 Mar 2010 08:47:43 -0600
Subject: [Microscopy] TEM liver

Contents Retrieved from Microscopy Listserver Archives
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Your tissue is too big, 1x1x1mm max.
Smaller is better, for dense tissues.

Greetings,

Joachim


J.Siegmund
Image Analyst
Seventhwave Labs
St. Louis

-----Original Message-----
X-from: dsoren-at-umich.edu [mailto:dsoren-at-umich.edu]
Sent: Thursday, March 04, 2010 7:45 AM
To: Joachim Siegmund

Hello all,

Does anyone have a tried and true protocol for processing liver for
TEM? We seem to be having problems with poor fixation. We have been
perfusing with 3% p.f. and 2.5 % glutaraldehyde in Sorensen's buffer,
followed by immersion fixation in the same mixture. We post fix with
1 % osmium in the same buffer, dehydrate with a graded series of
ethanol, change to propylene oxide, and then infiltrate and embed in
Epon. The cells immediately adjacent to the blood vessels and right
on the cut edge of the tissue are well fixed, but the cells located
on the interior of the tissue exhibit little white holes. The tissue
had been cut to about 1x2x2 mm. Another problem that we have
encountered in the past is cleared areas that remind me of canals
running through the cytoplasm. Is this to be expected? We would
love to hear from advice from some liver experts out there.

Thanks,

Dotty


Dorothy Sorenson
Microscopy and Image-analysis Laboratory
Department of Cell and Developmental Biology
University Of Michigan Medical School
A807 BSRB
109 Zina Pitcher Place
Ann Arbor, MI 48109-2200
(734)763-1170
FAX (734)763-1166



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20, 30 -- From jsiegmund-at-7thwavelabs.com Thu Mar 4 08:47:42 2010
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From: stefan.diller-at-t-online.de
Date: Thu, 4 Mar 2010 09:00:37 -0600
Subject: [Microscopy] Looking for LEICA EM AFS

Contents Retrieved from Microscopy Listserver Archives
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I am looking for a used LEICA EM AFS or EM AFS II.
If there is one for a reasonable price, please contact me offline.

Thanks,
Stefan


--
-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

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From: oshel1pe-at-cmich.edu
Date: Thu, 4 Mar 2010 10:21:33 -0600
Subject: [Microscopy] Fwd: Ask-A-Microscopist

Contents Retrieved from Microscopy Listserver Archives
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This is a good request for the micromavens at the natural history museums.

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
==========================================================

} Date: Thu, 4 Mar 2010 07:15:02 -0800 (PST)
} From: Mario Pino {mariopino-at-uach.cl}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Thursday, March 04,
} 2010 at 07:14:57 AM.
}
} realname - Mario Pino
} Email - mariopino-at-uach.cl
} ORGANIZATION - UACH Valdivia Chile
} EDUCATION - Graduate College
} LOCATION - Valdivia
} SUBJECT_OF_QUESTION - hair?
} QUESTION - we found a fossil of a gomphotere from the genus
} Stegomastodon that remeber a piece of skin. Under electron
} microscopy we found something thst remeber a hair. Can I send you a
} image to obtain some comemsts about it?
}
} regards
} m pino
}
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576
Microscopy Society of America
Ask a Microscopist
http://www.microscopy.org/microscopy/ask.cfm

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From: jflaci-at-ms.sapientia.ro
Date: Thu, 4 Mar 2010 16:04:17 -0600
Subject: [Microscopy] Re: Teeth again... and sample holder question...

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

Hello JQuinn!

It's a '80-s JSM-5200 from JEOL.
It has 10mm, 20mm and 48mm predefined working distances.
I tried to go down to 5mm, but then i was not able to focus the image.

Laci

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From: slc6-at-lehigh.edu
Date: Thu, 4 Mar 2010 20:50:54 -0600
Subject: [Microscopy] viaWWW: Scanning Probe Microscopy Short Course

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Email: slc6-at-lehigh.edu
Name: Sharon Coe

Organization: Lehigh University

Title-Subject: [Filtered] Scanning Probe Microscopy

Message: Scanning Probe Microscopy:
X-from Fundamentals to Advanced Applications
June 14-17, 2010
Lehigh University
Bethlehem, PA

This course provides an understanding of the
concepts, instrumentation, and applications of
the rapidly expanding field of Scanning Probe
Microscopy (SPM). Atomic Force Microscopy (AFM)
and Scanning Tunneling Microscopy (STM) will be
covered extensively, but the course will also
feature a variety of advanced SPM techniques.
The theory of operation for both imaging and
spectroscopy will be addressed, with special
attention being paid to instrument artifacts and
methods to avoid them. Crucially, the course
will include nearly equal time spent in
instruction and hands-on labs. Participants will
therefore take away the necessary knowledge and
practice to utilize the full potential of SPM
systems for applications in the physical and
biological sciences, including:

ïPractical and theoretical aspects of AFM and STM operation
ïForce-distance measurements (AFM indentation,
Force-Volume, molecular interactions,
instrumented Nano-indentation)
ïMechanical mapping (Phase imaging, multiple/higher order harmonics)
ïFluid measurements (Living cell work, Electrochemistry)
ïElectric field/surface potential imaging
ïMagnetic field imaging
ïPiezo-force microscopy (PFM)
ïNanolithography
ïImage Analysis
ïSystem management and probe selection

Instructors: Nancy Burnham (Worcester Polytechnic
Institute), Bryan Huey (University of
Connecticut),
Bruce Koel (Lehigh University), Dmitri Vezenov
(Lehigh University), Richard Vinci (Lehigh
University)

Participating Companies: Agilent Technologies,
Asylum Research, NT-MDT, Veeco Instruments

Each registrant receives the textbook, Scanning
Probe Microscopy and Spectroscopy: Theory,
Techniques, and Applications 2nd Ed., by Dawn
Bonnell (Wiley 2000), as well as detailed
laboratory notes for all hands-on exercises. In
addition, everyone receives notes for specific
lectures, a list of vendors and equipment
suppliers, and the Lehigh DVD that contains free
and demonstration versions of useful microscopy
software. Sample probes from several vendors are
also provided with support from probe
manufacturers.

For more information contact:
Sharon Coe (610-758-5133)
Sharon.coe-at-lehigh.edu
www.lehigh.edu/microscopy



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From: naomi_mccallum-at-health.qld.gov.au
Date: Thu, 4 Mar 2010 20:55:24 -0600
Subject: [Microscopy] Fwd: Re: automated light microscope slide scanner

Contents Retrieved from Microscopy Listserver Archives
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Dear Tom

One point to consider before you go too far down this path is to determine what the images will be used for. The assessment by Pathologists of some Aperio images is that the resolution is sometimes inadequate for pathological diagnosis.

Another that you have not listed is Olympus dotSlide which I have heard about but not trialed. Our department is more interested in realtime telepathology which is a slightly different ballgame.

regards
Naomi

Naomi McCallum
Supervising Scientist, EM Unit, Anatomical Pathology
Pathology Queensland
_________________________________________________
Clinical and Statewide Services Division| Queensland Health


} } } {mcauliff-at-umdnj.edu} 5/03/2010 12:52 am } } }



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We had the slide set (~180 slides) we use for teaching histology to
medical students scanned by Aperio rather than buy a scanner.
I suppose it all depends on how much demand you have for the service.

Geoff

PhillipsT-at-missouri.edu wrote:
} ----------------------------------------------------------------------------
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}
} I have been asked by a PI on our campus to consider adding an automated microscope slide scanner (e.g., Aperio, ImageExpress, NanoZoomer, etc) to our campus core facility. I am unsure of the market demand. Does anyone who man=
} ages a LM imaging core care to comment on this approach? Or someone who uses one routinely? I am also considering using an automated stage driven by metamorph to do this. I would just like to hear from others about how they approach this. Tom
}
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://www.biology.missouri.edu/faculty/phillips.html
} http://www.biotech.missouri.edu/mcc/
}
}
}
}
} ==============================Original Headers==============================
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From: litaduraine-at-earthlink.net
Date: Fri, 5 Mar 2010 08:09:54 -0600
Subject: [Microscopy] viaWWW: OTOTO artifacts

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Email: litaduraine-at-earthlink.net
Name: Lita Duraine

Organization: Biological Electron Microscopist

Title-Subject: [Filtered] OTOTO artifacts

Message: Hi Pete,

I used the OTOTO method for inner ears from toadfish while working at
NASA Ames Research Center extensively. The only difference was that
I used a CPD to dry the samples. I've always had good results as
long as the samples were fresh. Good Luck.

Lita Duraine
litaduraine-at-earthlink.net



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From: eschumacher-at-mccrone.com
Date: Fri, 5 Mar 2010 09:05:17 -0600
Subject: [Microscopy] Meeting Announcement: Midwest Microscopy and Microanalysis Society

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Greetings All,

The Midwest Microscopy and Microanalysis Society will hold its first meeting of 2010 on Friday, March 26th, at Northwestern University in Evanston, IL. The topic is "Super-Resolution Microscopy and Live-Cell Imaging". A preliminary program and registration information can be found on our website under Meetings:

http://midwestmicroscopy.org

Please join us to hear an excellent group of speakers and to show support for your Local Affiliate Society! We look forward to seeing you at this and future 2010 meetings.

Regards,

Elaine Schumacher
M3S Program Coordinator

*********************************************************************
Elaine F.Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com



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From: rbeavers-at-mail.smu.edu
Date: Fri, 5 Mar 2010 13:17:55 -0600
Subject: [Microscopy] Looking for 3D Laser Confocal Microscope Lab

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Group,

Trying to find a lab with a 3D Laser Confocal Microscope similar to the Zeiss LSM 700 in the North Texas area.

Student PHD project needs to build a detailed 3D model of small rodent teeth so precise measurements of tooth geometry can be made.

If you have an instrument, would be interested in availability and cost of use.

Thanks

Roy Beavers
Southern Methodist University
Department of Earth Sciences
P.O. Box 750395
Dallas, TX  75275
Voice: 214-768-2756
Fax: 214-768-2701
Email: rbeavers-at-smu.edu



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From: DusevichV-at-umkc.edu
Date: Fri, 5 Mar 2010 14:51:36 -0600
Subject: [Microscopy] RE: Teeth again... and sample holder question...

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Laci,

My greatest concern is that you stomatologist will treat dry tooth,
while in reality (with the exception of an enamel) tooth is pretty wet.
All current dental systems are designed by manufacturers for a wet
application. If stomatologist is not interested is junctions, like an
enamel-dentin junction, then cracks should not be of concern - you
always can find crack-free place for taking pictures, and anyway, cracks
mostly occur on junctions.

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784


} -----Original Message-----
} From: jflaci-at-ms.sapientia.ro [mailto:jflaci-at-ms.sapientia.ro]
} Sent: Thursday, March 04, 2010 1:01 AM
} To: Dusevich, Vladimir
} Subject: [Microscopy] Teeth again... and sample holder question...
}
}
}
}
}
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}
} Hello all!
}
} After the help You kindly provided for
} selecting the proper coating for teeth, i do have another problem.
}
} 1. One of my stomatologysts (former classmate in highschool) would
} like
} to take micrographs from tooth surface, than treat that surface
} than
} again micrographs an treat and so on, for 7-8 times.
}
} My problem is, that after 2 or 3 rounds of vacuuming in the SEM,
} the
} tooth will crack, and the whole thing is compromised, as the
} treatment material gets into the cracks and alters the result.
}
} Can any of You give me some advise how to avoid the cracks?
}
} 2. Does anybody have a 10mm WD sample holder for Jeol JSM-5200?
} If possible, I would like to kindly ask You to take some
} photographs
} so I could build my own. We have a pretty good machine shop at
} the
} University, they can handle this kind of work.
}
} Best regards,
} Laci
}
} PS:Of course coating is already not an option, but thanks anyway for
} the
} help.
} I will do the imaging at low kV(1-5) and small wd(10mm the minimum of
} the
} instrument).
}
}
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From: tjj-at-stowers.org
Date: Fri, 5 Mar 2010 17:16:32 -0600
Subject: [Microscopy] viaWWW: Job opening EM Specialist

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Email: tjj-at-stowers.org
Name: Teri Johnson

Organization: Stowers Institute for Medical Research

Title-Subject: [Filtered] Job opening EM Specialist

Message: The Stowers Institute has an opening for an Electron
Microscopy (EM) Specialist in the Histology Department. For more
information on this listing and to apply, see our website.
http://www.stowers.org/ScientistsSought/ScientistsSought.asp

Thanks,

Teri Johnson
Managing Director Histology Facility
Stowers Institute for Medical Research
Kansas City, MO

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From: david.r.hull-at-nasa.gov
Date: Fri, 5 Mar 2010 17:17:05 -0600
Subject: [Microscopy] viaWWW: Leica Ultracut UCT Preventive Maintenance - Ohio

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From: nizets2-at-yahoo.com
Date: Mon, 8 Mar 2010 06:48:32 -0600
Subject: [Microscopy] TEM liver

Contents Retrieved from Microscopy Listserver Archives
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Hi Dotty!

Apart from concentration, time and temperature are important factors to consider anywhere in science.
Try cutting small blocks and extending the fixation time (at 4°C).
On a side note, take care that PF is freshly prepared.

Best regards,

Stephane


----- Original Message ----
X-from: "dsoren-at-umich.edu" {dsoren-at-umich.edu}
To: nizets2-at-yahoo.com
Sent: Thu, March 4, 2010 2:39:11 PM

Hello all,

Does anyone have a tried and true protocol for processing liver for 
TEM?  We seem to be having problems with poor fixation.  We have been 
perfusing with 3% p.f. and 2.5 % glutaraldehyde in Sorensen's buffer, 
followed by immersion fixation in the same mixture.  We post fix with 
1 % osmium in the same buffer, dehydrate with a graded series of 
ethanol, change to propylene oxide, and then infiltrate and embed in 
Epon.  The cells immediately adjacent to the blood vessels and right 
on the cut edge of the tissue are well fixed, but the cells located 
on the interior of the tissue exhibit little white holes.  The tissue 
had been cut to about 1x2x2 mm.  Another problem that we have 
encountered in the past is cleared areas that remind me of canals 
running through the cytoplasm.  Is this to be expected?  We would 
love to hear from advice from some liver experts out there.

Thanks,

Dotty


Dorothy Sorenson
Microscopy and Image-analysis Laboratory
Department of Cell and Developmental Biology
University Of Michigan Medical School
A807 BSRB
109 Zina Pitcher Place
Ann Arbor, MI  48109-2200
(734)763-1170
FAX (734)763-1166



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From: hioag1-at-gmail.com
Date: Mon, 8 Mar 2010 09:46:20 -0600
Subject: [Microscopy] optical microscopy and mineral grain size characterization

Contents Retrieved from Microscopy Listserver Archives
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Greetings All,

I work at a mineral sands processing facility where cassiterite and
columbite are produced, and would like to better understand the
behavior of these minerals. It would be useful for me to:

1) Visually observe the mineral grains in sand type grab samples,
where the induvidual grains are about 10 - 1000 micrometers in size,
and

2) Take a digital photograph of 50 - 250 grains in a group, use some
type of software, and characterize the grains by size averages and
distributions.

Am presently focused on learning about the differences in
stereoscopes, Greenough and CMO, and having a difficult time deciding
which one is most suitable (if any) for this type of application,
ditto for cameras and software.

Any advice would be greatly appreciated.

Ed

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From: ahmad_ds-at-yahoo.com
Date: Mon, 8 Mar 2010 18:50:43 -0600
Subject: [Microscopy] viaWWW: SEM of polymer nano-filler composite

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Email: ahmad_ds-at-yahoo.com
Name: Ahmad Ashkaibi

Organization: Al-Balqa Applied University

Title-Subject: [Filtered] SEM of polymer nano-filler composite

Message: Hello everybody,

I've been asked to examine a polypropylene-nano-kaolinite composite
under SEM for particle distribution and size analysis. I am using a
30-kv FEI high-vacuum SEM. I've sputter-coated the polymer composite
specimens with gold. I've failed to get a good sharp image at high
magnification of nano-scale, no matter how low the kv or how small
the spot of the beam were. I just need to know if it's really
possible to examine nano-scale particles within a polymer sample on
SEM, or not. If anybody has been through the same headache and would
like to share their experience with us, it'd be extremely appreciated.

Thank you all very much.

Ahmad Ashkaibi

Materials and Metallurgical Engineering Department
Faculty of Engineering
Al-Balqa Applied University

As-Salt - 19117- Jordan

Email: ahmad_ds-at-yahoo.com
or: ashkaibi-at-bau.edu.jo

Tel.: 00962 7 77913343
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From: yamawaki-at-stanford.edu
Date: Mon, 8 Mar 2010 18:51:39 -0600
Subject: [Microscopy] viaWWW: LSRA Position- for Electron Microscopist

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Email: yamawaki-at-stanford.edu
Name: Ruth Yamawaki

Title-Subject: [Filtered] Job listing

Message: I am posting this for a PI. Please reply to her at the address below.

Stanford University LSRA Position- for Electron Microscopist

The Department of Comparative Medicine seeks a Life Science Research
Assistant (LSRA), to assist in research looking at neuronal
ultrastructural changes in the mammalian brain following experimental
manipulation. The LSRA will be joining a small dynamic neurobiology
laboratory and will be responsible for projects involving
transmission electron microscopy, including the preparation of
tissue, and its collection and analysis. Ultrastructural
immunocytochemistry will also be necessary. The position will be
part time (50% or higher) initially, with the potential to increase
to 100%.



Qualifications

Requires a Bachelor of Science degree in biology or related
scientific area, or higher. Must have demonstrable electron
microscopy experience, and skill in all aspects of electron
microscopy in mammalian central nervous system or brain tissue. They
must be highly organized with excellent communication and computer
skills, and have an ability to work independently as well as in a
team. Additional graphics, photography and statistical skills would
be advantageous.



To Apply

Please send CV, cover letter, and names of 3 references who may be
contacted, by email to Dr Corinna Darian-Smith at
cdarian-at-stanford.edu. Please only apply if you have demonstrable EM
experience. Salary will be commensurate with experience.



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From: bozzola-at-siu.edu
Date: Mon, 8 Mar 2010 19:02:48 -0600
Subject: [Microscopy] Re: viaWWW: SEM of polymer nano-filler composite

Contents Retrieved from Microscopy Listserver Archives
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I'm sure you tried this already. If not try:

1. low kV, uncoated specimen, secondary detector

2. high kV, carbon coated specimen, backscatter detector

Let us know what works (or not).

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

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From: gary-at-gaugler.com
Date: Mon, 8 Mar 2010 19:26:29 -0600
Subject: [Microscopy] Re: viaWWW: SEM of polymer nano-filler composite

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

Cannot recommend Au coating for nanoscale work. It coats as
a web rather than a film in most coaters. Thus, localized
charging--which degrades resolution ability. Sputter coating
with Pt or Ir is much better. Even better with high vacuum
turbo coater. If SEM has TLD, use that at 10KV, 2-3mm WD, spot 3,
and if UHR mode is available, use that. What SEM model do
you have? If Schottky SEM, it should work. If not, I doubt it.

gary g.


At 04:52 PM 3/8/2010, you wrote:



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From: protrain-at-emcourses.com
Date: Mon, 8 Mar 2010 19:42:21 -0600
Subject: [Microscopy] Re: viaWWW: SEM of polymer nano-filler composite

Contents Retrieved from Microscopy Listserver Archives
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Hi Ahmad

Whilst I agree with some of what Gary sets out I would caution the very
short working distance suggested for through the lens detection.

If it is charge that is causing you problems I would suggest 2kV and 7 -9mm
working distance, to increase the level of SE3 which would help reduce
charge effects.


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com]
Sent: 09 March 2010 01:27
To: protrain-at-emcourses.com

Cannot recommend Au coating for nanoscale work. It coats as
a web rather than a film in most coaters. Thus, localized
charging--which degrades resolution ability. Sputter coating
with Pt or Ir is much better. Even better with high vacuum
turbo coater. If SEM has TLD, use that at 10KV, 2-3mm WD, spot 3,
and if UHR mode is available, use that. What SEM model do
you have? If Schottky SEM, it should work. If not, I doubt it.

gary g.


At 04:52 PM 3/8/2010, you wrote:



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From: gary-at-gaugler.com
Date: Mon, 8 Mar 2010 20:12:49 -0600
Subject: [Microscopy] Re: viaWWW: SEM of polymer nano-filler composite

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

E-T cannot resolve nano particles on an FEI...or any other SEM.
TLD, UHR or in-lens is required...IMO and experience.

Why do you differ on the short WD? This is where the in-lens,
TLD and UHR have the highest resolution. If their FEI is
an SFEG, a non-Ferrous specimen ought not be an issue here.

I have not found E-T of nano specimens to be possible on
FEI or other SEMs at large WDs and in general, at short WD.
It requires an in-lens detector.

1-1.5nm resolution at 5-15KV using the in-lens detector at
2-3mm WD works. My specimens are SWCNT and MWCNT and by themselves,
do not present charging issues. But in a polymer, it is definitely
an issue. Therefore, the coating must be very fine and even
or the specimen is masked by the coating. This will cause
loss of resolution. It is a difficult situation. But it can
be done.

IMO, the low KV resolution of the FEI SFEG is not as good as
the Zeiss Gemini. Even so, the low KV resolution of the Zeiss
won't produce the resolution needed, depending on the size of
the particles. 30-60nm CNTs in the Zeiss will resolve at 1-2KV
but not in the FEI. Try to do this and see what you get and
compare.

First thing is to get rid of the Au coating and use a finer grain metal.
It is tricky since at high resolution, one can see the artifacts of
the coating but not resolve the actual specimen. No coating at
low KV and short WD with in-lens does work.

Disclaimer: I don't represent FEI or Zeiss. I'm just a user.

gary g.


At 05:43 PM 3/8/2010, you wrote:



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From: protrain-at-emcourses.com
Date: Tue, 9 Mar 2010 08:55:33 -0600
Subject: [Microscopy] Re: viaWWW: SEM of polymer nano-filler composite

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

Hi Gary

My advice re longer working distance was aimed at any problems the operator
may have which related to charge. So often the problem of "resolution" is
minimal on a modern TLD but what spoils the image is charge, or coating to
remove the charge. True I have not used the FEI but the Zeiss, JEOL and
Hitachi all respond to this technique.

Just information.


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com


-----Original Message-----
X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com]
Sent: 09 March 2010 02:14
To: protrain-at-emcourses.com

E-T cannot resolve nano particles on an FEI...or any other SEM.
TLD, UHR or in-lens is required...IMO and experience.

Why do you differ on the short WD? This is where the in-lens,
TLD and UHR have the highest resolution. If their FEI is
an SFEG, a non-Ferrous specimen ought not be an issue here.

I have not found E-T of nano specimens to be possible on
FEI or other SEMs at large WDs and in general, at short WD.
It requires an in-lens detector.

1-1.5nm resolution at 5-15KV using the in-lens detector at
2-3mm WD works. My specimens are SWCNT and MWCNT and by themselves,
do not present charging issues. But in a polymer, it is definitely
an issue. Therefore, the coating must be very fine and even
or the specimen is masked by the coating. This will cause
loss of resolution. It is a difficult situation. But it can
be done.

IMO, the low KV resolution of the FEI SFEG is not as good as
the Zeiss Gemini. Even so, the low KV resolution of the Zeiss
won't produce the resolution needed, depending on the size of
the particles. 30-60nm CNTs in the Zeiss will resolve at 1-2KV
but not in the FEI. Try to do this and see what you get and
compare.

First thing is to get rid of the Au coating and use a finer grain metal.
It is tricky since at high resolution, one can see the artifacts of
the coating but not resolve the actual specimen. No coating at
low KV and short WD with in-lens does work.

Disclaimer: I don't represent FEI or Zeiss. I'm just a user.

gary g.


At 05:43 PM 3/8/2010, you wrote:



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From: jacques.faerber-at-ipcms.u-strasbg.fr
Date: Tue, 9 Mar 2010 10:10:11 -0600
Subject: [Microscopy] Re: viaWWW: SEM of polymer nano-filler composite

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

Hi

Maybe it's not a SEM or parametring issue, but a preparation one ?
How does the sample look like, before any preparation ? Do you cut it or
brake it for examining the inside, or do you look at it external surface ?
From my experience on such composites, the external surface doesn't
reflect what is inside. In particular, one see bad the talc, the
kaolinite, the TiO2 charges of the composite. I would brake the sample,
and to have a brittle fractured surface, it must be made at low temperature.

You dip the sample in liquid nitrogen until the nitrogen doesn't boil
any more, you take it out and you brake it fast, before it goes up in
temperature, with a hammer blow on an anvil. The pieces will flow
everywhere in the room...
OK, it's a bit a brutal way to prepare samples, not much reproducible,
but it works. (If someone knows a better way to do the same, I'm
interested to hear about...).

You'll look at the broken surface of such a piece, and you schould be
able to see somthing from the different compounds. If it looks like
melted cheese, it means that the sample was not cold/hard enough
anymore. Try to be faster when you get it out of the nitrogen !

Of coarse, as other said, you have to use no coating, or carbon, or less
Ir, Os, or Pt and no gold, use low energy (1-5 keV), and try with both
ET or TTL detectors.

Hope it helps.

J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr



ahmad_ds-at-yahoo.com a écrit :
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} Email: ahmad_ds-at-yahoo.com
} Name: Ahmad Ashkaibi
}
} Organization: Al-Balqa Applied University
}
} Title-Subject: [Filtered] SEM of polymer nano-filler composite
}
} Message: Hello everybody,
}
} I've been asked to examine a polypropylene-nano-kaolinite composite
} under SEM for particle distribution and size analysis. I am using a
} 30-kv FEI high-vacuum SEM. I've sputter-coated the polymer composite
} specimens with gold. I've failed to get a good sharp image at high
} magnification of nano-scale, no matter how low the kv or how small
} the spot of the beam were. I just need to know if it's really
} possible to examine nano-scale particles within a polymer sample on
} SEM, or not. If anybody has been through the same headache and would
} like to share their experience with us, it'd be extremely appreciated.
}
} Thank you all very much.
}
} Ahmad Ashkaibi
}
} Materials and Metallurgical Engineering Department
} Faculty of Engineering
} Al-Balqa Applied University
}
} As-Salt - 19117- Jordan
}
} Email: ahmad_ds-at-yahoo.com
} or: ashkaibi-at-bau.edu.jo
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From: oshel1pe-at-cmich.edu
Date: Tue, 9 Mar 2010 10:40:55 -0600
Subject: [Microscopy] Re: viaWWW: SEM of polymer nano-filler composite

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

I just hold the sample with pliers. And try not
to break the pliers (which I've done).
Works a treat.
Phil

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From: xyang-at-SMU.CA
Date: Tue, 9 Mar 2010 11:01:12 -0600
Subject: [Microscopy] RE: viaWWW: SEM of polymer nano-filler composite

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Hi Ahmad,

I assume you have already adjusted the SEM into its best performance by
using a resolution standard.

If I am right, your real problem is how to get a clear image of nano-scale
particles "within" a polymer sample -- that is -- both the primary and
secondary e-beams have to penetrate the polymer film in order for you to
obtain the particle information. The image will thus for sure be blurry, not
matter how hard you try.

The solutions I can think of are: 1. completely dissolve the polymer
material and view the remaining nanoparticles under the SEM. 2. break the
polymer composite and get a fresh surface so that the particles are exposed
to air.

Good luck and Best regards,

Xiang (Sean) Yang
Regional Analytical Centre
Saint Mary's University
Halifax, NS Canada B3H 3C3
Phone: 902-420-5709
http://fgsr.smu.ca/emc/
http://www.smu.ca/institutes/rgc/welcome.html



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From: meshin-at-umich.edu
Date: Tue, 9 Mar 2010 12:26:46 -0600
Subject: [Microscopy] TEM of RBC

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Hello Everybody,
I have been asked to do TEM of red blood cell membrane which, I have
never done that before. I really appreciate if you have any advice,
suggestion, or protocols that you can share it with me.

Thank You very Much
Sasha Meshinchi

---------------------------------
Sasha Meshinchi
The University of Michigan
Cell & Developmental Biology
Microscopy & Image-analysis Lab
A830 BSRB
Ann Arbor, MI 48109-0616
Phone: (734) 763-1170 (lab)
Fax: (734) 763-1166
E-mail: meshin-at-umich.edu
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From: ahmad_ds-at-yahoo.com
Date: Tue, 9 Mar 2010 20:04:36 -0600
Subject: [Microscopy] viaWWW:Thanks - Re: SEM of polymer nano-filler composite

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Email: ahmad_ds-at-yahoo.com
Name: Ahmad Ashkaibi

Organization: BAU

Title-Subject: [Filtered] Re: SEM of polymer nano-filler composite

Message: Thank you all for your responses.

Special thanks to Gary , for his useful notes.
Although, I maybe should have mentioned that I'm using an E-T
detector, and I've got no other option. Besides, the gun is tungsten
filament, so I concluded from your replies that it's impossible to
see the nano particles at these conditions.

The coating available is either gold or carbon, so I guess that's
another problem.

Thank you Emily for your comments about the size of clay. I've
fractured the polymer sample at room temperature and coated the
fracture surface.
The "supposed to be" nano kaolintine has been added after re-melting
the PP pellets in a double screw hot blender (BRABENDER).

Thank you very much.



Ahmad Ashkaibi

Electron Microscopy Unit
Materials and Metallurgical Engineering Department
Faculty of Engineering
Al-Balqa Applied University

As-Salt - 19117- Jordan

Email: ahmad_ds-at-yahoo.com
or: ashkaibi-at-bau.edu.jo

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From: pwf-at-fullam.com
Date: Tue, 9 Mar 2010 20:05:16 -0600
Subject: [Microscopy] viaWWW: Free Books

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Email: pwf-at-fullam.com
Name: Dianne Fullam

Organization: Fullman

Title-Subject: [Filtered] Free Books

Message:
Hi all,

We have the following books available for the cost of shipping:

Microscopy and Micro analysis Proceeding 2002, 2004, 2005, 2006, 2008

Fracture, An Advanced Treatise Volume 1 Microscopic and Macroscopic
Fundamentals c 1968

Index to the X-ray Powder Data File c 1960

Ultrasoft X-ray Microscopy: Its Application to Biological & Physical
Sciences, Annals of the NY Academy of Sciences

vol 342 c 1980

The Effective Use and Proper Care of the Microscope by Oscar W. Richards c

1949

The Toxicology of Beryllium by US Dept of Health, Education and Welfare c

1972

Preparation Techniques for TEM by Zeiss

The Basics of Materials for Thin Films by the Materials Research
Corporation.

Ultrasonic and Their Scientific and Technical Applications by Bergman and
Hatfield c 1943

Handbook of Non-Ferrous Metallurgy by Liddell Vol 1& 2 c 1926

Factor Analysis in Chemistry by Maunowski 1980

ASTM Standards Part 31Physical and Mechanical Testing of Metals;
Nondestructive Tests, May 1968

ASTM Standards Part 32 Chemical Analysis of Metals: Sampling and Analysis of
Metal Bearing Ores, May 1968

ASTM Standards General Test Methods Part 30 July 1971 and Part 30 July 1973

Nuclear Fuels - Beckerly, Gurinsky, & Dienes 1956

Qualitative Analysis by Long, Chamberlain and Anderson c 1926

ASRC Report 1977-78, Atmospheric Research Center, SUNY Albany

Graphic Arts Techniques and Equipment, NASA c 1970

Solar Probe Helios c 1974

Photo Lab Index, The Cumulative Formulary of Standard Recommended
Photographic Procedures by Henry M Lester c 2 vols

Diffusion of Tin Cladding in the Rolling of Lead Foil, Thesis for the Master
Of Metallurgical Engineering by Murray H Levine 1960

An Electron-Optical Study of Certain Lamellar Mineral Preparations:

Exfoliated Graphite and Flocculated Kaolin

Thesis of Richard E. Stevens 1975

American Men Of Science No. 1 The Physical Sciences 9th ed. c 1955 1 volume

American Men of Science, The Physical and Biological Sciences, 10th ed. c.

1960 4 volumes

Van Nostrand's Scientific Encyclopedia 5th ed. 1976

Random House Encyclopedia 1983



Please contact us at sales-at-fullam.com with your shipping address and which
books you are interested in. We will reply with the shipping cost payable by
check or credit card.

Dianne Fullam

Ernest F. Fullam, Inc



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From: John.Catino-at-Mineralstech.com
Date: Wed, 10 Mar 2010 10:12:01 -0600
Subject: [Microscopy] Re: viaWWW: SEM of polymer nano-filler composite

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Ahmad,

Plasma etching the polymer will expose particles.


John

John W. Catino
Group Leader Analytical Services
Microscopy & Organic Characterization
Specialty Minerals, Inc.
MINTEQ International, Inc.
640 N. 13th Street
Easton, PA 18042

M: 610.533.6766
F: 610.250.3344
john.catino-at-mineralstech.com




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m
To
03/08/10 07:52 PM john.catino-at-mineralstech.com
cc

Please respond to Subject
ahmad_ds-at-yahoo.co [Microscopy] viaWWW: SEM of polymer
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Email: ahmad_ds-at-yahoo.com
Name: Ahmad Ashkaibi

Organization: Al-Balqa Applied University

Title-Subject: [Filtered] SEM of polymer nano-filler composite

Message: Hello everybody,

I've been asked to examine a polypropylene-nano-kaolinite composite
under SEM for particle distribution and size analysis. I am using a
30-kv FEI high-vacuum SEM. I've sputter-coated the polymer composite
specimens with gold. I've failed to get a good sharp image at high
magnification of nano-scale, no matter how low the kv or how small
the spot of the beam were. I just need to know if it's really
possible to examine nano-scale particles within a polymer sample on
SEM, or not. If anybody has been through the same headache and would
like to share their experience with us, it'd be extremely appreciated.

Thank you all very much.

Ahmad Ashkaibi

Materials and Metallurgical Engineering Department
Faculty of Engineering
Al-Balqa Applied University

As-Salt - 19117- Jordan

Email: ahmad_ds-at-yahoo.com
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From: sbarlow-at-sunstroke.sdsu.edu
Date: Wed, 10 Mar 2010 11:24:23 -0600
Subject: [Microscopy] Cryosectioning with RMC CR-X unit

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Listers

I am trying to use an acquired RMC CR-X crysectioning module on a
Leica EM UC6 ultramicrotome. I am interested in discussing how this
unit works, in general, and any experiences users might have had with
the CR-X on a Leica microtome. Please feel free to contact me
offline, or respond to the list.

Thanks in advance

Steve
--
Dr. Steven Barlow
EM Facility/Biology Dept.
San Diego State University
5500 Campanile Drive MC-4614
San Diego CA 92182-4614
phone: (619) 594-4523
fax: (619) 594-5676

email: sbarlow-at-sunstroke.sdsu.edu
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From: ralkhat-at-ncsu.edu
Date: Wed, 10 Mar 2010 18:39:51 -0600
Subject: [Microscopy] viaWWW: Freeze Fracture Supplies

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Email: ralkhat-at-ncsu.edu
Name: Rami Alkhatib

Organization: NCSU

Title-Subject: [Filtered] Freeze Fracture Supplies

Message: Dear All,
Does any body know where to find/buy freeze fracture sample prep.
supplies? We are looking for stainless steel specimen holding basket
that fits inside a 500 ml LN2 dewar. Also, we are looking for the
gold alloy specimen carrier (studs) for mounting the samples.

I highly appreciated if anyone can guide me where to find these items.

P.S. I searched in Leica, EMS, fisher scientific..etc.

Thanks!

Rami Alkhatib, Ph.D



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From: dlowry-at-asu.edu
Date: Wed, 10 Mar 2010 18:40:15 -0600
Subject: [Microscopy] viaWWW: TEM sample preparation for culture cells

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Name: David Lowry

Organization: Arizona State University

Title-Subject: [Filtered] TEM sample preparation for culture cells

Message: I would like to ask for advice on preparing mammalian
culture cells for thin-section TEM that have been grown in
suspension. We plan to use standard chemical fixation methods but how
to handle the cells--can they be treated like bacteria and pelleted
into agarose following primary fixation, or are there other
approaches that have been used successfully? We would like to avoid
repeated pelleting during processing. Thank you-

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From: dac-at-research.umass.edu
Date: Wed, 10 Mar 2010 20:18:38 -0600
Subject: [Microscopy] Re: viaWWW: Freeze Fracture Supplies

Contents Retrieved from Microscopy Listserver Archives
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Hi Rami,

You didn't mention what brand of FF instrument you are working with. Let
us know.... it probably matters.

If it is a Balzers unit, Balzers/Baltec supplies are now handled by
Leica. Contact Todd Perez at Leica. Todd.Perez-at-leica-microsystems.com

Some supplies are more general and there are alternate suppliers. The
Balzers film thickness monitors can use crystals from several suppliers.
We've had good luck with Tangidyne crystals.

Dale



ralkhat-at-ncsu.edu wrote:
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} Email: ralkhat-at-ncsu.edu
} Name: Rami Alkhatib
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} Title-Subject: [Filtered] Freeze Fracture Supplies
}
} Message: Dear All,
} Does any body know where to find/buy freeze fracture sample prep.
} supplies? We are looking for stainless steel specimen holding basket
} that fits inside a 500 ml LN2 dewar. Also, we are looking for the
} gold alloy specimen carrier (studs) for mounting the samples.
}
} I highly appreciated if anyone can guide me where to find these items.
}
} P.S. I searched in Leica, EMS, fisher scientific..etc.
}
} Thanks!
}
} Rami Alkhatib, Ph.D
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8, 20 -- From dac-at-research.umass.edu Wed Mar 10 20:18:38 2010
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From: ralkhat-at-ncsu.edu
Date: Wed, 10 Mar 2010 21:39:48 -0600
Subject: [Microscopy] viaWWW: Freeze fracture supplies for Cressington CFE-60

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Name: Rami Alkhatib

Organization: NCSU

Title-Subject: [Filtered] Freeze fracture supplies for Cressington CFE-60

Message: Hi Dale,
We are using Cressington freeze fracture (Cressington CFE-60).

Thanks!




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From: skperkin-at-vt.edu
Date: Thu, 11 Mar 2010 08:41:55 -0600
Subject: [Microscopy] viaWWW: Printer for TEM working prints

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Email: skperkin-at-vt.edu
Name: Sandy Hancock

Organization: Virginia Tech

Title-Subject: [Filtered] Printer for TEM working prints

Message: Good Morning,

I realize that this topic has been discussed previously, but printers
have come and gone since the last discussion thread. What are labs
currently using to print quality TEM working prints on plain paper?
Most interested in a printer cost {$1000.

Thank you,
Sandy Hancock



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From: Elliott-at-arizona.edu
Date: Thu, 11 Mar 2010 10:53:40 -0600
Subject: [Microscopy] Re: viaWWW: TEM sample preparation for culture cells

Contents Retrieved from Microscopy Listserver Archives
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Low Melting Point Agarose is good. One can suspend the cells in LMP,
spin them, and then put the tube on ice to harden.

Another option is to soft fix the cells, re-suspend them in 0.5ml of
0.1M CaCo + 8% BSA and pellet. Gently add 4 drops of 25% Glut (do not
disturb the pellet) and let the glut hard fix your cells and cross-
link the BSA (about 45 min).

Then, cut off the cell containing pellet and cut into small bits. No
more spinning needed.

I have used both methods. Each have advantages and disadvantages. In
each case, you can control the speed/length of the final spin to
either maximize cell density or cell shape.

David


On Mar 10, 2010, at 5:43 PM, dlowry-at-asu.edu wrote:

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} Title-Subject: [Filtered] TEM sample preparation for culture cells
}
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} culture cells for thin-section TEM that have been grown in
} suspension. We plan to use standard chemical fixation methods but how
} to handle the cells--can they be treated like bacteria and pelleted
} into agarose following primary fixation, or are there other
} approaches that have been used successfully? We would like to avoid
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From: kristi.majni-at-basf.com
Date: Thu, 11 Mar 2010 18:23:07 -0600
Subject: [Microscopy] viaWWW: EM on sodium bifluoride

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Email: kristi.majni-at-basf.com
Name: Kristi Majni

Organization: BASF

Title-Subject: [Filtered] EM on sodium bifluoride

Message: Hello all,

I recently had a request to take some images of some sodium
bifluoride powders. Sodium bifluoride is extremely corrosive and when
heated to 125C, gives off HF. I'm asking the list's opinions on
whether or not this would harm the column of my Hitachi S-3500N. I
expect to have many requests in the future for this stuff, so would
long-term exposure be a problem?

Thank you!

Kristi Majni

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From: kenconverse-at-qualityimages.biz
Date: Thu, 11 Mar 2010 18:57:51 -0600
Subject: [Microscopy] viaWWW: EM on sodium bifluoride

Contents Retrieved from Microscopy Listserver Archives
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Hi Kristi
I'm not familiar with sodium bifluoride, but I would think that proper
notification of any field service personnel, every visit, and proper
notification to anyone who might rebuild any of your vacuum pumps, or even
just dispose of the oils, would be imperative. 125¢ªC would not be hard to
achieve with an e-beam and HF is not to be messed with, particularly
unknowingly.

Just my $.02 from a service perspective.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
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Name: Kristi Majni

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Title-Subject: [Filtered] EM on sodium bifluoride

Message: Hello all,

I recently had a request to take some images of some sodium
bifluoride powders. Sodium bifluoride is extremely corrosive and when
heated to 125C, gives off HF. I'm asking the list's opinions on
whether or not this would harm the column of my Hitachi S-3500N. I
expect to have many requests in the future for this stuff, so would
long-term exposure be a problem?

Thank you!

Kristi Majni

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From: gary-at-gaugler.com
Date: Thu, 11 Mar 2010 19:22:43 -0600
Subject: [Microscopy] Re: viaWWW: EM on sodium bifluoride

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NaHF2 is poisonous, corrosive and decomposes when heated.
If this is to be a recurring observation, consider requiring
a cooling stage or specimen holder to maintain the specimen
at perhaps -25C. It seems to me that if the material is
repeatedly studied and heated, the chamber, column and metal parts
will probably be corroded and eventually need replacement.

I would suggest reviewing an MSDS for this material and
ask some pointed questions. Also, see the advisories for
handling this material.

gary g.



At 04:24 PM 3/11/2010, you wrote:



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From: nizets2-at-yahoo.com
Date: Fri, 12 Mar 2010 02:47:02 -0600
Subject: [Microscopy] Re: viaWWW: TEM sample preparation for culture cells

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

Interesting approach, I take good note.
However, I usually just pellet the cells once in a conical tube and then fix them and the pellet is hard enough to be treated as it is.
The specimen never moves from the tubes until curing of the resin and I add the solutions gently against the tube walls to avoid dislocation of the pellet.
After osmium fixation the pellets are pretty hard.

Best regards,

Stephane



----- Original Message ----
X-from: "Elliott-at-arizona.edu" {Elliott-at-arizona.edu}
To: nizets2-at-yahoo.com
Sent: Thu, March 11, 2010 5:57:13 PM

Low Melting Point Agarose is good.  One can suspend the cells in LMP, 
spin them, and then put the tube on ice to harden.

Another option is to soft fix the cells, re-suspend them in 0.5ml of 
0.1M CaCo + 8% BSA and pellet.  Gently add 4 drops of 25% Glut (do not 
disturb the pellet) and let the glut hard fix your cells and cross-
link the BSA (about 45 min).

Then, cut off the cell containing pellet and cut into small bits.  No 
more spinning needed.

I have used both methods.  Each have advantages and disadvantages.  In 
each case, you can control the speed/length of the final spin to 
either maximize cell density or cell shape.

David


On Mar 10, 2010, at 5:43 PM, dlowry-at-asu.edu wrote:

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} Title-Subject: [Filtered] TEM sample preparation for culture cells
}
} Message: I would like to ask for advice on preparing mammalian
} culture cells for thin-section TEM that have been grown in
} suspension. We plan to use standard chemical fixation methods but how
} to handle the cells--can they be treated like bacteria and pelleted
} into agarose following primary fixation, or are there other
} approaches that have been used successfully? We would like to avoid
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From: oshel1pe-at-cmich.edu
Date: Fri, 12 Mar 2010 08:26:51 -0600
Subject: [Microscopy] flat-bed scanners as off spring 2010

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

Listers,

This question has been asked several times before, yes, but (as with
the printer question), manufacturers are always changing things and
bringing out new models. So:
As of Spring 2010, what is the best flat-bed scanner for digitizing
3.25 X 4 TEM negatives? (And preferably also does other size
negatives, documents, etc.).

We currently have an Epson Perfection 4990 Photo. Works well, but I
just had to fix it, and it's old, so I expect the bulbs to go before
long.
Looking on line, I find the Epson V500 Photo. Specs look good, but ... ?

Plus of course The Economy, and we're looking at serious budget cuts,
so price matters.
Thanks.

Phil
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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From: rjharris-at-uwo.ca
Date: Fri, 12 Mar 2010 10:49:53 -0600
Subject: [Microscopy] flat-bed scanners as off spring 2010

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

Hi Phil
Based on feedback from the list I purchased the Espon V700 last year (the
V500 is more of an office scanner) and have been very pleased with the
output. I scan 6 TEM negatives at a time so going through a camera's worth
of film is not onerous

Rick,

Richard Harris, Manager - Integrated Microscopy -at- Biotron
The Biotron - Center for Experimental Climate Change Research
University of Western Ontario,
London Ontario, CANADA.
N6A 5B7
Ph.  519-661-2111 ext. 86780
Fax  519-661-3935
e-mail rjharris-at-uwo.ca
web: www.thebiotron.ca

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Friday, March 12, 2010 9:36 AM
To: rjharris-at-uwo.ca

Listers,

This question has been asked several times before, yes, but (as with
the printer question), manufacturers are always changing things and
bringing out new models. So:
As of Spring 2010, what is the best flat-bed scanner for digitizing
3.25 X 4 TEM negatives? (And preferably also does other size
negatives, documents, etc.).

We currently have an Epson Perfection 4990 Photo. Works well, but I
just had to fix it, and it's old, so I expect the bulbs to go before
long.
Looking on line, I find the Epson V500 Photo. Specs look good, but ... ?

Plus of course The Economy, and we're looking at serious budget cuts,
so price matters.
Thanks.

Phil
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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5, 23 -- From: Philip Oshel {oshel1pe-at-cmich.edu}
5, 23 -- Subject: flat-bed scanners as off spring 2010
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From: Elliott-at-arizona.edu
Date: Fri, 12 Mar 2010 11:33:28 -0600
Subject: [Microscopy] flat-bed scanners as off spring 2010

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

I second Rick's suggestion. I am very happy with my V700 for TEM negs
and manny other things. I must admit that many personal 35mm slides
have been done by this scanner also :-)
David


On Mar 12, 2010, at 9:54 AM, rjharris-at-uwo.ca wrote:

}
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} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi Phil
} Based on feedback from the list I purchased the Espon V700 last
} year (the
} V500 is more of an office scanner) and have been very pleased with the
} output. I scan 6 TEM negatives at a time so going through a
} camera's worth
} of film is not onerous
}
} Rick,
}
} Richard Harris, Manager - Integrated Microscopy -at- Biotron
} The Biotron - Center for Experimental Climate Change Research
} University of Western Ontario,
} London Ontario, CANADA.
} N6A 5B7
} Ph. 519-661-2111 ext. 86780
} Fax 519-661-3935
} e-mail rjharris-at-uwo.ca
} web: www.thebiotron.ca
}
} -----Original Message-----
} X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
} Sent: Friday, March 12, 2010 9:36 AM
} To: rjharris-at-uwo.ca
} Subject: [Microscopy] flat-bed scanners as off spring 2010
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Listers,
}
} This question has been asked several times before, yes, but (as with
} the printer question), manufacturers are always changing things and
} bringing out new models. So:
} As of Spring 2010, what is the best flat-bed scanner for digitizing
} 3.25 X 4 TEM negatives? (And preferably also does other size
} negatives, documents, etc.).
}
} We currently have an Epson Perfection 4990 Photo. Works well, but I
} just had to fix it, and it's old, so I expect the bulbs to go before
} long.
} Looking on line, I find the Epson V500 Photo. Specs look good,
} but ... ?
}
} Plus of course The Economy, and we're looking at serious budget cuts,
} so price matters.
} Thanks.
}
} Phil
} --
} Philip Oshel
} Microscopy Facility Supervisor
} Biology Department
} 024C Brooks Hall
} Central Michigan University
} Mt. Pleasant, MI 48859
} (989) 774-3576
}
} ==============================Original
} Headers==============================
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5, 22 -- From Elliott-at-arizona.edu Fri Mar 12 11:33:27 2010
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From: hstahlberg-at-me.com
Date: Fri, 12 Mar 2010 11:53:55 -0600
Subject: [Microscopy] Re: flat-bed scanners as off spring 2010

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


Dear Rick and David,
Would you have any data for the scanner's point spread function? If it
goes up to 6400 dpi, or about 4 micrometers per pixel, what is the
scanner's PSF or MTF?
If you scan a grey-value Kodak TEM negative, do you see adjacent
pixels going "black-white-black-white", with other words, does the
scanner reach Nyquist frequency at all?
How reliable is focussing?
The data are recorded with CCD technology. Are the grey-value data 8
bit, or 16 bit, or somewhere in-between?
Thanks,
Henning.


Henning Stahlberg
Center for Cellular Imaging and Nano Analytics (C-CINA)
at the Department of Biosystems Science and Engineering (D-BSSE)
Structural Biology and Biophysics, Biozentrum,
WRO-1058, Mattenstrasse 26
University Basel, CH-4058 Basel, Switzerland
Tel: +41-61-387 32 62
mailto:Henning.Stahlberg-at-unibas.ch
http://c-cina.org





On Mar 12, 2010, at 6:44 PM, Elliott-at-arizona.edu wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
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} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} I second Rick's suggestion. I am very happy with my V700 for TEM negs
} and manny other things. I must admit that many personal 35mm slides
} have been done by this scanner also :-)
} David
}
}
} On Mar 12, 2010, at 9:54 AM, rjharris-at-uwo.ca wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------------
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} }
} } Hi Phil
} } Based on feedback from the list I purchased the Espon V700 last
} } year (the
} } V500 is more of an office scanner) and have been very pleased with
} } the
} } output. I scan 6 TEM negatives at a time so going through a
} } camera's worth
} } of film is not onerous
} }
} } Rick,
} }
} } Richard Harris, Manager - Integrated Microscopy -at- Biotron
} } The Biotron - Center for Experimental Climate Change Research
} } University of Western Ontario,
} } London Ontario, CANADA.
} } N6A 5B7
} } Ph. 519-661-2111 ext. 86780
} } Fax 519-661-3935
} } e-mail rjharris-at-uwo.ca
} } web: www.thebiotron.ca
} }
} } -----Original Message-----
} } X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
} } Sent: Friday, March 12, 2010 9:36 AM
} } To: rjharris-at-uwo.ca
} } Subject: [Microscopy] flat-bed scanners as off spring 2010
} }
} }
} }
} }
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} } ----------------------------------------------------------------------------
} }
} } Listers,
} }
} } This question has been asked several times before, yes, but (as with
} } the printer question), manufacturers are always changing things and
} } bringing out new models. So:
} } As of Spring 2010, what is the best flat-bed scanner for digitizing
} } 3.25 X 4 TEM negatives? (And preferably also does other size
} } negatives, documents, etc.).
} }
} } We currently have an Epson Perfection 4990 Photo. Works well, but I
} } just had to fix it, and it's old, so I expect the bulbs to go before
} } long.
} } Looking on line, I find the Epson V500 Photo. Specs look good,
} } but ... ?
} }
} } Plus of course The Economy, and we're looking at serious budget cuts,
} } so price matters.
} } Thanks.
} }
} } Phil
} } --
} } Philip Oshel
} } Microscopy Facility Supervisor
} } Biology Department
} } 024C Brooks Hall
} } Central Michigan University
} } Mt. Pleasant, MI 48859
} } (989) 774-3576
} }
} } ==============================Original
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} } 5, 23 -- From oshel1pe-at-cmich.edu Fri Mar 12 08:26:51 2010
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==============================Original Headers==============================
11, 23 -- From hstahlberg-at-me.com Fri Mar 12 11:53:55 2010
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From: mbrown-at-aaechighschools.com
Date: Fri, 12 Mar 2010 14:05:20 -0600
Subject: [Microscopy] Schematics for Cambridge S200 motorized stage controller

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

My controller board is dead and the schematics do not appear in the
standard S200 book of circuit diagrams. The schematics would greatly
aid in its resuscitation.

In fact if you have grown tired of your S200 and wish its stage
controller board would go away, I could help you out with that :-)

-Dr. Mike Brown
Science Dept Chair
AAEC-PV



==============================Original Headers==============================
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From: donk-at-ardl.com
Date: Fri, 12 Mar 2010 15:47:21 -0600
Subject: [Microscopy] RE: Cambridge S200 Motorized Stage Controller

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

Mike, Try Secondary Images at: http://www.secondaryimages.com/ . Clark should have some parts for that item.

Don Kierstead
Microscopist
ARDL, Inc.
www.ardl.com
(866) 778-ARDL [toll free]
(330) 794-6600 [worldwide]
(330) 794-6610 [fax]



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From: arrowood-at-utep.edu
Date: Fri, 12 Mar 2010 22:45:35 -0600
Subject: [Microscopy] viaWWW: EM on sodium bifluoride

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Email: arrowood-at-utep.edu
Name: Roy Arrowood

Organization: University of Texas at El Paso

Title-Subject: [Filtered] EM on sodium bifluoride

Message: If powder particle shape and size are the main topics of
interest, you might be able to meet the client needs by doing TEM of
replicas, rather than putting the actual material in the instrument.

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From: jhun.yew-at-gmail.com
Date: Sun, 14 Mar 2010 09:55:32 -0500
Subject: [Microscopy] viaWWW: EDX penetration depth program

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Email: jhun.yew-at-gmail.com
Name: jhun

Organization: Nanyang Technology University

Title-Subject: [Filtered] EDX penetration depth program

Message: Would like to ask if anyone knows of a shareware program
that is able to calculate the EDX penetration depth. From what i was
told it is the shareware program which goes by the name of casino,
but an online search returned with 'other' results as one would
imagine.

Any help would be most appreciated

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From: kenconverse-at-qualityimages.biz
Date: Sun, 14 Mar 2010 12:44:32 -0500
Subject: [Microscopy] viaWWW: EDX penetration depth program

Contents Retrieved from Microscopy Listserver Archives
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Jhun,
If you add electron penetration to the search you'll find
http://www.gel.usherbrooke.ca/casino/What.html
I think this is what you're looking for.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


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Email: jhun.yew-at-gmail.com
Name: jhun

Organization: Nanyang Technology University

Title-Subject: [Filtered] EDX penetration depth program

Message: Would like to ask if anyone knows of a shareware program
that is able to calculate the EDX penetration depth. From what i was
told it is the shareware program which goes by the name of casino,
but an online search returned with 'other' results as one would
imagine.

Any help would be most appreciated

Login Host: 202.156.10.228
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From: donovan-at-uoregon.edu
Date: Sun, 14 Mar 2010 12:45:25 -0500
Subject: [Microscopy] Re: viaWWW: EDX penetration depth program

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

Hi Jhun,
You can download Casino from my company's Technical page along with a
number of other freeware programs useful for microscopy:

http://www.probesoftware.com/Technical.htm

The Casino download link is at the bottom of the page.
john

At 08:01 AM 3/14/2010, jhun.yew-at-gmail.com wrote:



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From: Henrik.Kaker-at-guest.arnes.si
Date: Sun, 14 Mar 2010 14:36:46 -0500
Subject: [Microscopy] Re: viaWWW: EDX penetration depth program

Contents Retrieved from Microscopy Listserver Archives
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Please see the link, http://www2.arnes.si/~sgszmera1/html/monte_carlo.html.

Best regards,

Henrik


jhun.yew-at-gmail.com wrote:
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}
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}
} Title-Subject: [Filtered] EDX penetration depth program
}
} Message: Would like to ask if anyone knows of a shareware program
} that is able to calculate the EDX penetration depth. From what i was
} told it is the shareware program which goes by the name of casino,
} but an online search returned with 'other' results as one would
} imagine.
}
} Any help would be most appreciated
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--
Dr. Henrik Kaker
Ob Suhi 23
SI-2390 Ravne
Slovenia
GSM: +386 31 784 281
http://www.kaker.com
Email:info-at-kaker.com


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From: mmiralles-at-pi.ac.ae
Date: Mon, 15 Mar 2010 06:55:32 -0500
Subject: [Microscopy] Mineral Identification Software

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

Hi,

In relation to mineral identification, is there an available software
that can aid mineral identification using the Quant values from the EDS?
Let's say, if I have around 58% F, 33% Ca and 7% C.

Will appreciate an input.

Thanks,
Melina


==============================Original Headers==============================
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5, 29 -- Subject: Mineral Identification Software
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From: Frank_Karl-at-lincolnelectric.com
Date: Mon, 15 Mar 2010 07:36:06 -0500
Subject: [Microscopy] Re: Mineral Identification Software

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

At the risk of putting my foot in my mouth...
Since so many element compositions have the possibility of several
different crystal structure which defines them as minerals, I would have
say that without additional optical or X-ray crystal data you couldn't
truly identify a material as a specific mineral.

I submit chrysotile and lizardite. Both have the same chemical
composition, but without knowing that one is fibrous and the other is
blocky you could not separate them.

EDS data can get you part way, but in my opinion you need some additional
information to push you over.



mmiralles-at-pi.ac.a
e
To
03/15/2010 08:03 frank_karl-at-lincolnelectric.com
AM cc

Subject
Please respond to [Microscopy] Mineral Identification
mmiralles-at-pi.ac.a Software
e












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Hi,

In relation to mineral identification, is there an available software
that can aid mineral identification using the Quant values from the EDS?
Let's say, if I have around 58% F, 33% Ca and 7% C.

Will appreciate an input.

Thanks,
Melina


==============================Original
Headers==============================
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From: Michal.Jarnik-at-fccc.edu
Date: Mon, 15 Mar 2010 08:30:40 -0500
Subject: [Microscopy] Need to move an old Ultracut E

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

Due to space limitations in the lab I need to get rid of an old (25+
years) Reichert Ultracut E microtome with a FC4 cryoattachment. I will
through in the pump and dewar as well. I used it last time about 5 years
ago, that time it was working both for plastic and cryo, but I cannot
offer any guarantees.

The instrument is located in Philadelphia PA, whoever grabs it is
responsible for packaging and shipping. Local pick-up preferred. Please
contact me off-list if interested.

Thanks, Michal

--




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From: pe_fischione-at-fischione.com
Date: Mon, 15 Mar 2010 08:46:43 -0500
Subject: [Microscopy] viaWWW: Job opening: Applications Scientist

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

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://microscopy.com/MLFormMail.html
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Email: pe_fischione-at-fischione.com
Name: Paul E. Fischione

Organization: E.A. Fischione Instruments, Inc

Title-Subject: [Filtered] Job opening: Applications Scientist

Message: E.A. Fischione Instruments, Inc. is a
highly-successful and innovative scientific
instrumentation company looking to expand its
world-class team with the following position:

Applications Scientist
This position supports the sales force from a
technical perspective in interactions with
customers and potential customers. It requires
being a self-starter with excellent verbal and
written communication skills.
Qualifications and Experience:
ï Are you an individual who works harder
than most people and thrives in a fast-paced
environment?
ï Do you respond positively and enthusiastically to challenges?
ï Do you take pride in your work and hate to make mistakes?
Responsible for full customer support of both
Fischione and TVIPS GmbH products. TVIPS is a
major manufacturer of cameras and software for
transmission electron microscopy (TEM).

With time, needs to develop an acute knowledge of
both Fischioneís and TVIPSí products as well as
competitive devices and techniques. Required to
participate in sales calls and trade shows.
Performs advanced product demonstrations as
necessary. Responsible for full customer field
support, including independent installation,
on-site customer training, telephone/email
support, and hardware and software
troubleshooting of new and existing products.
Responsible for ensuring that products meet
operating specifications, performs documented
service, and manages scheduling and logistics.

Maintains microscopy facilities and is
responsible for procedures, documentation, and
equipment performance. Performs original
research and publishes in peer-reviewed journals.
Participates in technical conferences and
contributes presentations or posters. Promotes
companyís image and products through technical
forums and application notes. Creates and
ensures the technical accuracy of any corporate
literature or publication. Frequently
collaborates with Fischioneís end users.

Requires a Ph.D. in Physics, Engineering,
Materials Science, Biology or a related subject
and strong transmission electron microscopy (TEM)
experience for various life and physical science
applications. Experience with TEM camera
technology, digital imaging, and electron
tomography is beneficial. Must be willing to
travel between 25% and 40%, both domestically and
internationally.

Individuals willing to work in a fast-paced
environment, going through rapid growth and
change are encouraged to apply. Please visit our
web site at www.fischione.com to learn more about
E.A. Fischione Instruments, Inc.

E.A. Fischione Instruments, Inc. is an Equal
Opportunity Employer committed to diversity. We
offer competitive salaries and benefits.


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From: Michal.Jarnik-at-fccc.edu
Date: Mon, 15 Mar 2010 09:11:31 -0500
Subject: [Microscopy] Ultracut E moved

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

Thanks for interest, the microtome found its new home. Michal



==============================Original Headers==============================
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From: marie.cantino-at-uconn.edu
Date: Mon, 15 Mar 2010 10:34:50 -0500
Subject: [Microscopy] job opening, University of Connecticut

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

The Department of Physiology and Neurobiology at the University of
Connecticut, Storrs is seeking to fill a position for a Research
Assistant II/III in the Electron Microscopy Laboratory.
Responsibilities include, but are not limited to, repair and
maintenance of equipment, training and supervision of equipment users,
general laboratory maintenance, and coordination of building related
repairs. This is a full-time, annually renewable position. Go to http://www.hr.uconn.edu/employment_services/jobs-fac.html#2010275
for qualifications.

Screening of applications will begin immediately and will continue
until the position is filled. Preference will be given to
applications received before April 2, 2010.

In keeping with our commitment to build a culturally diverse
community, the University of Connecticut invites applications from
women, people with disabilities, and members of minority groups.
(Search #2010275)



Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369


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From: Michal.Jarnik-at-fccc.edu
Date: Mon, 15 Mar 2010 11:10:53 -0500
Subject: [Microscopy] Lubricants for a Nikon Hg lamp housing

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

A few of our Hg lamps have a problem with sticking alignment of the lamp
and especially the mirror. I took the mechanism apart (and even put it
back together :), pretty simple) and it seems it would need cleaning and
fresh lubricant. The temperatures in that part of the housing are quite
high and I am not sure what to use. Any smart ideas anybody?

Thanks, Michal

--


==============================Original Headers==============================
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From: john_mackenzie-at-ncsu.edu
Date: Mon, 15 Mar 2010 13:38:34 -0500
Subject: [Microscopy] Re:flat-bed scanners as off spring 2010

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

Folks:
This question has come up before and I have additional data to add.
Let me preface first by stating that my laboratory is not involved in
rigorous testing of images for three dimensional image reconstruction
where I personally believe another level of precision is achieved/required.
I was curious because all of my measurements of the the Epson
4880/4990/D750M, D750M pro indicated that Epson produced images of TEM
negatives scans that were CLEARLY superior to everybody elses (including
older Epson scanners. This was based on examining details of TEM and
comparing them under a microscope with actual photographs of the same
negative printed in a darkroom on a Durst enlarger with Agfa Brovira
film. (For you younger guys film is the stuff we used to use in the dark
ages to record images. The darkroom equipment was "the best"). From this
point forward dots per inch which is a meaningless term, will be
replaced exclusively by pixels per inch.
Although careful one to one examination showed that every detail was
retained, indicating the grayscale resolution and spatial resolution was
sufficient), there was no doubt that a trained microscopist could pick
out the darkroom printed images every time. "They were sharper" (This is
not a quantifiable entity. Before you jump all over me read on)
I decided to "calibrate" my scanner. As noted in the previous post, one
the best measures of the resolution of the system is the MTF. To make an
eight month quest short, Don Wilson at Kodak would wrote the ISO
standard on scanner resolution helped me measure the MTF. It was
difficult to believe that a flatbed scanner that cost $800 was beating
the resolution of drum scanners that cost $27,000- 100,000. These
scanners however had been rigorously tested by "disciples" from the MRC
and were known to produce excellent data.
Film "experts (photographers some of whom were engineers made logical
blunders and confused the MTF of the film with the MTF of the camera)(
see work of Ken Taylor's group on the resolution of film by diffraction
criteria) The "best" photographers argue whether the resolution of film
is 4000 pixels per inch or 8000 pixels per inch.
There are clear answers to this issue that I should be writing up for
Microscopy Today but are not necessary to understand right now. The
important point is that consensus is greater than 4000 pixels per inch.
(Don't bother with your arguments about microns/pixel, etc. I know, I
know .. not now)
So I went out and purchased $3,400 worth of targets and "standards" and
measured the scanners MTF. I thought I would a hard quantifiable number
that was "unbiased" by human variability. Nope.
The MTF requires the user to define the noise limit that is "acceptable"
for discerning gray levels. Photographers have a range that is pretty
well defined because images that fall below a certain contrast level
"look" too grainy and therefore are clearly "unacceptable" BUT a
scientist/microscopist looking at that image sees that it is noisy but
wouldn't reject it until the niose level was two, three, or four times
worse (in rare cases, even worse than that ) Additionally the MTF is
dependent on the size of the scanned pixel and there are real issues
with the number of grayscale values required and the size of the scanned
pixel. (This is a question of whether we are looking at the resolution
of the film as defined by it's ability to resolve diffraction data or 12
bit gray scale data. I think a films spatial resolution is higher when
defined by diffraction critera than by 12 bit imaging criteria. (This
opens yet another can of worms)
Finally, (I love this one) the MTF measures contrast and frequency. It
does not and cannot measure sharpness. whoops. What measures sharpness.
not edges cause the MTF measures edges to get contrast and frequency.
The photographers use that ultra precise measuring tool called the
photographer and he/she says, " Well it looks sharper" The examples
shown in the majority of cases would never pass scientific scrutiny.
My tests were based on the assumption that I needed more than eight bits
so I chose twelve bits per pixel as my necessary lower limit.
Using a target for scanner reflective image resolution measurement based
on contrast versus resolvable lines per inch (which can be
mathematically converted to pixels per inch.
The target showed that all the scanners listed above could achieve 2000
pixels per inch resolution. This is the upper limit of all the targets
including the USAF target and the ISO target that is commercially available.
I had two 4990's that were measured and surprisingly one passed one did
not. The one that didn't was far worse in one axis versus the other. but
both failed.
Working with Don Wilson at Kodak (who by the way authored ALL the iso
specs), we measured all at a little better than 2000 pixels per inch
when using a noise level I figured was acceptable to most microscopists.
His measurements showed clearly and quantifiabley that one 4990 was broken
Those of you who are really paying attention will note that I said
reflective and we are more concerned about transparency scanning. In the
Epsons transparency scanning is not the same as reflective scanning. So
one should test transparency scanning in the same manner. Good luck.
There are NO targets available. Seems there is ZERO demand. I really
mean ZERO. I had some long discussions with the manager at the company
the makes the reflective target and he agreed to make me a custom target
for transparency. (I understood that it might have a minor problem with
certification as a "standard" but I knew that it could tell me if it
coulds see 2000 pixel per inch in transparency mode in both directions
and that that would work for me.
Got the target and all passed but the broken one. Same with the MTF all
passed but the broken one.
Here is my take on the data above.
Specifications listed by the manufacturers are total bull, have no
scientific validity, do not pass any scientific test and are all subject
to human interpretation. (It is that noise ratio thing that I can't get
nailed down for TEM images as we would normally view them in
conventional stained embedded TEM.)
The MTF does not measure sharpness which as you might recall was the
clear difference between the dark room printed image and the scanned
and printed image.
Let me throw in another line of experimentation we did. We wanted to
know if an eight megapixel digital camera printing images at 19 inches
by 13 inches captured and printed images that were limited by the
resolution of either the camera's detector or by the printers
resolution. (Although there is an iso specification on scanning
resolution there has never been one done on printers. Three years ago
when I last spoke with Don he as working on it but he was concerned that
Kodak may not want to continue with that line of experimentation
(externally))
The answer was that we had to more than half the resolution before we
could even detect the effects of a sharpening filter without a microscope.
The final data that is available now comes from the photographic
experts. They don't agree on anything except. The really good people
believe that film can resolve 4000 pixels per inch OR 8000 pixels per
inch if scanned in a high quality drum scanner under liquid immersion.
(NEVER a flat bed)
My take on discussions with groups that scanned film and then used the
data for critical reconstructions was that they were closer to the 4000
to 8000 pixel per inch number. Ken Taylor group has I believe an even
higher number because the spots can achieve a higher resolution than a
12 bit grayscale image pixel. (I'm not going to debate this because it
isn't relevant to the topic here)
I think a vast majority of the experts that were using film for
reconstruction and were active in the measurement of it's limitations
are now using digital acquisition systems that increase their
productivity more than 10 fold over working manually. Because everything
can be automated, the productivity can skyrocket beyond that.. The
scientific fact that the detector can measure all the detail available
has been determined critically. However, because film has a much higher
resolution the field of view is decreased substantially. This fact is
less than irrelevent if you are collecting data at ten, a hundred or
possibly even more than you could prior to digital automation. The
reduction in field of view can be a problem for say a pathologist. The
reconstruction groups love that the detectors are linear. As an imaging
person I can tell you that this fact is the biggest PROBLEM with digital
systems but that's another major tome.
Pulling it together for the big climax.
When we printed negatives in the darkroom we could blow up the images
many fold. A 35 mm film can easily be blown up to 13 x 19. which is
greater than ten. Thats a minimum. By the same logic our film can
therefore be blown up 10 fold to 40in x 52. (3.25 x 4 times 13) We only
print at a maximum of 8 x 10 and much less for the journals. (The 40 in
x 52 in format is quite rare.) So we don't use all that film has. and
under most conditions we can therefore drop the resolution we need from
the discussion of the upper limit at 4000 ppi or 8000 ppi to the 2000
ppi that the scanner can actually do when working properly and operated
properly. (Now I would say calibrated properly but that is problematic.
The target that I got for measuring the MTF of the transparency scanner
was a gift and was produced on a classified, propriety scanner that is
no longer available.Because the Epson scanners use the same scanning
mechanism for both reflective and transparency scanning you can at least
infer that things are right ( My data suggests that there is a strong
correlation of the two and I have never seen the transparency messed up
without the reflective being messed up also. Doesn't mean it can't
happen but it is unlikely.
We are 99% convinced that the film has at least 12 bits of data. We scan
at 16, store at 16 and then convert to 8 for most imaging. Again another
tome but this has no controversy among the groups that have really
looked at the data.
We do nothing but scan negatives for all our digital. We scan routinely
at 1200 ppi, 16 bit grayscale full size and store as tif 16 bit. This
gets us most of the data that 8 x 10 would get us from the past. When we
want high resolution we are normally after a smaller selected area and
can afford to scan at the higher resolution.
The Epson scanners are the only scanners I even consider. Measurements
show that they get at least 2000 ppi and although that is not comparable
to film it is equivalent to a 4k by 4k digital camea. (do a price
comparison and the scanner has a slight advantage). Most TEMs in the
future will take most of their images by digital and less by film. Soon
we will go the same route that the fashion, ad, and high end
professional photographers and the detector will hit film resolution
and then go beyond it. ( Hey, it's only sand and technology after all.)
The price will fall and guys in the future will wonder what the heck
those people from the past were debating.(And what is that film stuff
anyway?)
As I am about to climb on my soapbox, dear readers who are satisfied can
stop reading.
One of the major problems that I see going forward is that students
actually think they know what they are doing. From the standpoint of
understanding digital imaging sufficiently to make critical ethical and
scientific decisions on what they can do and what they can't they are
positively clueless. Clueless people who think they know what they are
doing are bond to make serious mistakes. Is there increased interest in
properly educating them in the science of imaging? No Do students and
advisors think that students need this training ? No
With the increased use in digital data manipulation, how often are
programs tested for scientific validity? Do you know the answer to that
question? Are you using Photoshop? Do you understand that there is an
issue with how Adobe handles 16 bit numbers?
We use transparency scanners and yet no target is commercially available
to measure MTF (Please notify the list if you know a source)
I think that we actually need more data and more serious discussion as I
see it nearly impossible to clearly define what is ethic to do to an
image and what is unethical if you can't define the original data with
any degree of precision.
I stopped Chairing Symposia because too many individuals complained that
the topic had been beaten to death and that there was nothing new. I am
probably not going to give my Digital Imaging Course because students
and colleagues feel that the course is not necessary. I neither want to
Chair the Symposia nor teach the course but would ask that smeone
actually remember that we are doing science and that in science you need
controls and you need to understand how all the instruments that measure
things work. When you don't and you ASSUME you knoe how it works you are
headed for trouble.
Charlie Lyman cleverly put me on the Editorial Board of Microscopy Today
so the above should make at least a three part article in the coming
months . In addition, I am working on a handbook which includes much of
the practical workflow with scientific justification for the Sunday
Short Course. this year. (which will likely be my last) Now if I can
just stay healthy and alive long enough to complete thse tasks, I'll be
happy.
Sorry way too much soapbox but if you followed my directions you are not
reading this anyway.
Now, if this doesn't stir up a hornets nest of controversy, I will be
shocked. BUT please be advised that I didn't have enough time to write
this so that it is unlikely I'm going to get into a battle of verbal
jousting on the list server. Make sure that before you leap into this
you are clear about the MSA statement on ethical digital imaging. Since
the data is scientific data, there is little that you can do without
having to report your manipulations in detail. In addition, since it is
data, how you acquired that data must be understood and should in my
opinion be validated.

John Mackenzie

john_mackenzie-at-ncsu.edu

--

John Mackenzie, Jr. PhD

Coordinator for the Center for Electron Microscopy

Professor of Microbiology

North Carolina State University

Phone (919) 515-2664 Fax (919) 515-8293


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From: donald.gibbon-at-matcoinc.com
Date: Mon, 15 Mar 2010 14:20:02 -0500
Subject: [Microscopy] Re: Mineral Identification Software

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

Ditto squared! EDS quant is so often mis-used, it's almost a joke! It's
so easy to get numbers and so hard to get the right ones.

The first thing is not to believe anything that doesn't come off a
polished section... and to NEVER take a single value but to take
multiple scans over small areas (not points) and generate more
representative data. THEN you can begin to speculate on the mineral
identity. Make sure, of course, you're getting weight percent values
(not atomic percent) since most mineral compositions are represented by
weight. Then, of course, if you can x-ray the material you'll be far
more likely to have it right as far as the actual phase present.

Another thing to recognize is that most minerals are not stoichiometric
end members but are actually somewhere out in multi-dimensional phase
space... so don't be surprised if you don't get ideal values. And for
that example you gave with F, Ca and C... is the carbon in the mineral
or the mount?

I also recommend adding transmitted cross-polarized light microscopy to
add another traditional dimension to your investigations especially if
the minerals aren't opaque. Their optical properties can be enormously
helpful. This was the main ID tool for the hundred years before EDS
became the crutch most of us lean on. It's a bit like chemical
microscopy... almost forgotten and enormously powerful, fast and cheap.
And it doesn't depend on software at all!!! Jut your eyes and brain.

Donald L. Gibbon
MATCO Services

-----Original Message-----
X-from: Frank_Karl-at-lincolnelectric.com
[mailto:Frank_Karl-at-lincolnelectric.com]
Sent: Monday, March 15, 2010 8:42 AM
To: Gibbon, Donald L.

At the risk of putting my foot in my mouth...
Since so many element compositions have the possibility of several
different crystal structure which defines them as minerals, I would have
say that without additional optical or X-ray crystal data you couldn't
truly identify a material as a specific mineral.

I submit chrysotile and lizardite. Both have the same chemical
composition, but without knowing that one is fibrous and the other is
blocky you could not separate them.

EDS data can get you part way, but in my opinion you need some
additional
information to push you over.




mmiralles-at-pi.ac.a

e


To
03/15/2010 08:03 frank_karl-at-lincolnelectric.com

AM
cc



Subject
Please respond to [Microscopy] Mineral
Identification
mmiralles-at-pi.ac.a Software

e


















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

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


Hi,

In relation to mineral identification, is there an available software
that can aid mineral identification using the Quant values from the EDS?
Let's say, if I have around 58% F, 33% Ca and 7% C.

Will appreciate an input.

Thanks,
Melina


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From: jkrupp-at-deltacollege.edu
Date: Mon, 15 Mar 2010 17:09:21 -0500
Subject: [Microscopy] Denatured alcohol

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

So, I was planning a party and went to the solvent room to get the
makings. (Not really!)

When I got there all I found were jugs labeled Ethanol, denatured, CDA
19. I had been wondering about this, the alcohol in the lab had a
strange odor and sometimes did not seem to be the same as the 95% at
other jobs.

I found out that CDA 19 stands for Completely Denatured Alcohol
Formula 19. Formula 19 has things like methyl isobutyl ketone,
kerosene, or aviation gas mixed in so it is undrinkable. Well you can
drink it, you will get sick.

This is my first exposure to CDA 19 and I was wondering if anyone else
has run across this and if you have any thoughts on using CDA 19 in
place of traditional 95% ETOH for tissue processing.

Thanks

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu




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From: PhillipsT-at-missouri.edu
Date: Mon, 15 Mar 2010 18:39:39 -0500
Subject: [Microscopy] Denatured alcohol

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


Jon,

I have not seen bottles labeled with CDA 19 but I will look in the Pathology Lab to see if they have it.

Many years ago I learned to make all dilutions for ethanol dehydrations for TEM from 100% (99.9%) not like they do in the histology lab (LM) which used the 95% gallon jugs or bigger. I use older opened bottles for the lower percentages to save money and this works very well. The difference was quite noticeable.

Pat


Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E - Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-480-6560
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov} {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not represent the NIH. This message is not confidential and can be freely shared and reproduced.


________________________________
X-from: {jkrupp-at-deltacollege.edu}
Reply-To: {jkrupp-at-deltacollege.edu}

This doesn't address the tissue processing question but there was recently an interesting story in Slate online magazine of the government's intentional "denaturation" of alcohol during Prohibition and how it poisoned (and killed) hundreds/thousands of citizens. Check out this link:

http://www.slate.com/id/2245188
The Chemist's War
The little-told story of how the U.S. government poisoned alcohol during Prohibition with deadly consequences.

Tom


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: jkrupp-at-deltacollege.edu [mailto:jkrupp-at-deltacollege.edu]
Sent: Monday, March 15, 2010 5:10 PM
To: Phillips, Thomas E.

So, I was planning a party and went to the solvent room to get the
makings. (Not really!)

When I got there all I found were jugs labeled Ethanol, denatured, CDA
19. I had been wondering about this, the alcohol in the lab had a
strange odor and sometimes did not seem to be the same as the 95% at
other jobs.

I found out that CDA 19 stands for Completely Denatured Alcohol
Formula 19. Formula 19 has things like methyl isobutyl ketone,
kerosene, or aviation gas mixed in so it is undrinkable. Well you can
drink it, you will get sick.

This is my first exposure to CDA 19 and I was wondering if anyone else
has run across this and if you have any thoughts on using CDA 19 in
place of traditional 95% ETOH for tissue processing.

Thanks

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu




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From: slego-at-agraquest.com
Date: Mon, 15 Mar 2010 19:17:57 -0500
Subject: [Microscopy] viaWWW: Sample preparation bacteria resting on a leaf

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Email: slego-at-agraquest.com
Name: Sarah Lego

Organization: AgraQuest

Title-Subject: [Filtered] Sample preparation

Message: I am trying to image bacteria resting on
a leaf (not in a biofilm), but when I dye with
fluorescent stain/fix for SEM imagery, the
bacteria fall off into the liquid of the
stain/fixative and I am not able to capture them
resting on the leaf. I was curious how to best
prepare a sample such as this for fluorescence or
electron microscopy that perhaps wouldnít involve
liquid. Thank you very much.



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From: jtwilley-at-sprynet.com
Date: Mon, 15 Mar 2010 19:40:45 -0500
Subject: [Microscopy] Re: Denatured alcohol

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I can't speak for this specific designation but the chemical properties of the denaturant are a factor in some other fields as well. In art conservation (and to some extent in lacquer formulation) it is preferable for us to use a grade that has only other alcohols as denaturants (i.e., without ketones, petroleum distillates, etc.) One type that is readily available in hardware stores uses methanol and isopropanol as the denaturants and works better for solvating resins whose solubility is impaired by non-alcohols. I'm sure that laboratory grades of the same thing are available.

You might also consider the suppliers that serve the microelectronics industry, as they have very stringent standards and are a bit off the radar in terms of academic and bio labs.

John Twilley


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From: mmiralles-at-pi.ac.ae
Date: Tue, 16 Mar 2010 00:21:51 -0500
Subject: [Microscopy] RE: Mineral Identification Software

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Thank you for all your replies, they are all very helpful. I will check
the links/software posted.

The missing element to make up the 100% is Oxygen, by the way. I have
taken xray data from a freshly broken surface, on the flat part of the
cleavage. Though I am not sure of the mineral yet, I am sure that the C
came from the sample and not the mount. We are still checking if this is
a part of the original material or a result of leaching of some sort. We
only have a small piece of the sample that is why we are holding off
optical microscopy, if we can. I will review my method again based from
your inputs.

Thank you so much.

Melina





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From: benada-at-biomed.cas.cz
Date: Tue, 16 Mar 2010 05:17:26 -0500
Subject: [Microscopy] Re: viaWWW: Sample preparation bacteria resting on a leaf

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Hello Sarah,
I would suggest to try a fixation in OsO4 vapour. There is an old method for
preserving arial fungal structures published by King and Brown (1983). We are
using it for samples of arial Streptomycetes mycelia for SEM. It also works
well on fungal mycelium grown on conifera leafs.

Here is full citation:
King E. J., Brown M. F., 1983. A technique for preserving aerial fungal
structures for scanning electron microscopy. Can J Microbiol 29: 653-658.

Best regards from Prague
Oldrich

--
Oldrich Benada
Institute of Microbiology
Acad. Sci. CR
Videnska 1024
CZ-142 20 Prague 4
Czech Republic

On Tuesday 16 of March 2010 01:19:57 you wrote:
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} a leaf (not in a biofilm), but when I dye with
} fluorescent stain/fix for SEM imagery, the
} bacteria fall off into the liquid of the
} stain/fixative and I am not able to capture them
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From: Frank_Karl-at-lincolnelectric.com
Date: Tue, 16 Mar 2010 05:52:20 -0500
Subject: [Microscopy] Re: Denatured alcohol

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Jon,
When I started at rubber company in Akron we had 2B alcohol, a denatured
ethanol. It was denatured with 2% benzene. I heard stories about rubber
worker who would put raw rubber in the 2B alcohol to absorb the benzene and
would then drink what was left. They would get sick too, but not until
later...

stay safe.....
Frank




jkrupp-at-deltacolle
ge.edu
To
03/15/2010 06:18 frank_karl-at-lincolnelectric.com
PM cc

Subject
Please respond to [Microscopy] Denatured alcohol
jkrupp-at-deltacolle
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So, I was planning a party and went to the solvent room to get the
makings. (Not really!)

When I got there all I found were jugs labeled Ethanol, denatured, CDA
19. I had been wondering about this, the alcohol in the lab had a
strange odor and sometimes did not seem to be the same as the 95% at
other jobs.

I found out that CDA 19 stands for Completely Denatured Alcohol
Formula 19. Formula 19 has things like methyl isobutyl ketone,
kerosene, or aviation gas mixed in so it is undrinkable. Well you can
drink it, you will get sick.

This is my first exposure to CDA 19 and I was wondering if anyone else
has run across this and if you have any thoughts on using CDA 19 in
place of traditional 95% ETOH for tissue processing.

Thanks

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu




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From: klivi-at-jhu.edu
Date: Tue, 16 Mar 2010 09:57:24 -0500
Subject: [Microscopy] Re: Mineral Identification Software

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

Dear Melina,

I have to deal with the question you ask all the time in my
profession. In order to identify a mineral, I rely on three things: 1)
structure, 2) composition, 3) locality or source. Structure is the
least ambiguous and the easiest way to identify a mineral is by powder
X-ray diffraction. However, X-ray identification is made easier by
knowledge of the composition. In the case where you have very little
of the sample, single crystal precession X-ray photography can work
well, but not many people have that technique. The TEM can combine
diffraction and EDS on small samples, so it is my method of choice.
However, you have to consume part of the sample to do TEM.

In doing mineral ID, I always use knowledge of the source as a way to
narrow down the possibilities. I rarely use composition alone to
positively ID a mineral... especially from EDS data from an SEM. The
error in these analyses can be quite high and when you take that into
account, you may not be able to narrow down the possibilities. In
addition, solid solution of elements within many mineral structures
makes compositional ID difficult.

If you want to narrow the possibilities down, go to a mineral database
like: www.webmineral.com (not the only one). There you will find some
possible candidates. However, you will not find an exact match to your
composition. With your example, I would first guess fluorite CaF2,
although the C is problematic. Diffraction information on the TEM
could easily determine if the material is fluorite. (Watch out for
beam damage though!)

Mineral ID takes some experience and I don't think there can be a
quick look up list that will give you an easy answer just from
composition. To be unequivocal, you need diffraction data.

Hope this helps,
Ken


On Mar 15, 2010, at 7:58 AM, mmiralles-at-pi.ac.ae wrote:

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} Hi,
}
} In relation to mineral identification, is there an available software
} that can aid mineral identification using the Quant values from the
} EDS?
} Let's say, if I have around 58% F, 33% Ca and 7% C.
}
} Will appreciate an input.
}
} Thanks,
} Melina
}
}
} ==============================Original
} Headers==============================
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|_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
Kenneth JT Livi, PhD
The High-Resolution Analytical Electron Microbeam Facility of the
Integrated Imaging Center
Departments of Earth and Planetary Sciences and Biology
Olin Hall
3400 N Charles Street
Johns Hopkins University
Baltimore, Maryland 21218 USA
| | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| |
| :|








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From: DusevichV-at-umkc.edu
Date: Tue, 16 Mar 2010 10:35:28 -0500
Subject: [Microscopy] RE: viaWWW: Sample preparation bacteria resting on a leaf

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

You can try to minimize specimen preparation procedure: just air dry leaves and sputter coat them. You do not need stain for SEM and quite often air borne bacteria is visible without fixation.
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

} -----Original Message-----
} From: slego-at-agraquest.com [mailto:slego-at-agraquest.com]
} Sent: Monday, March 15, 2010 7:18 PM
} To: Dusevich, Vladimir
} Subject: [Microscopy] viaWWW: Sample preparation bacteria resting on a
} leaf
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} Title-Subject: [Filtered] Sample preparation
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} Message: I am trying to image bacteria resting on
} a leaf (not in a biofilm), but when I dye with
} fluorescent stain/fix for SEM imagery, the
} bacteria fall off into the liquid of the
} stain/fixative and I am not able to capture them
} resting on the leaf. I was curious how to best
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From: Frank_Karl-at-lincolnelectric.com
Date: Tue, 16 Mar 2010 10:58:23 -0500
Subject: [Microscopy] Mineral Identification Software

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

Please don't take this as an attack on anyone, I accept there are more than
one way to poach a egg, but:

If you think your sample is CaF2 an optical microscopist has faster and
easier ways to check it's identity. It's cubic xtal, so it has one
refractive index. I don't have my copy of Winchell's optical properties
but a quick internet search indicated the refractive index of CaF2 is N
(iso)=1.435. Mounting a few small xtals in water (n=1.33) and then
ethylene glycol (1.438) the light microscope will show the suspected CaF2
has a refractive index greater than 1.33 and less but almost matching
refractive index to 1.438. I'd expect to see a colored Becke line if the
dispersion values of ethylene glycol and xtal were close. With a little
more work and a table of two you could predict the color. Cross your polars
(if you haven't done it all ready) and the field is black and the xtals are
also at extinction over 360 degees of stage rotation.

If so we know our material is"
A: cubic or a glass (look for conchoidal fracture to separate those)
B: refractive index slightly less then 1.438
C: EDS found large amounts of Ca and F

Already I'd be thinking about the report I wanted to write.

So why am I taking your time up? Because I see a tendency to look at
numbers from a computer and accept them as Truth. I've see it in technical
people who say , "If EDS prints out 2 significant figures, then by Gawd it
must be right." I've seen it with people measuring alpha quartz in clay
without regard to peak interference. We need to back check our analysis by
means of a seperate methodology.

Thanks for the rant. Boy, do I miss optical microscopy



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10, 22 -- From frank_karl-at-lincolnelectric.com Tue Mar 16 10:58:23 2010
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10, 22 -- To: Microscopy-at-microscopy.com
10, 22 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005
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From: klivi-at-jhu.edu
Date: Tue, 16 Mar 2010 11:18:04 -0500
Subject: [Microscopy] Re: Mineral Identification Software

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

Dear Frank,
You're right that refractive index could give you another way to
identify the grain. Pretty cheaply too! Unless you don't have the oils
etc. I tend to rely on the methods available to me. And I agree that
numbers can be misleading. I think I emphasized that. When someone
wants to prove that they have found a new mineral, both X-ray
diffraction and refractive index is normally required. However, the
latter requirement has been relaxed somewhat (I believe) in that not
all new minerals can be physically separated or large enough to have
the index measured. When given the choice between index and
diffraction data, I personally chose diffraction. That's just me (with
not statement that we should abandon the oils)!
Cheers!
Ken

On Mar 16, 2010, at 12:01 PM, Frank_Karl-at-lincolnelectric.com wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Please don't take this as an attack on anyone, I accept there are
} more than
} one way to poach a egg, but:
}
} If you think your sample is CaF2 an optical microscopist has faster
} and
} easier ways to check it's identity. It's cubic xtal, so it has one
} refractive index. I don't have my copy of Winchell's optical
} properties
} but a quick internet search indicated the refractive index of CaF2
} is N
} (iso)=1.435. Mounting a few small xtals in water (n=1.33) and then
} ethylene glycol (1.438) the light microscope will show the suspected
} CaF2
} has a refractive index greater than 1.33 and less but almost matching
} refractive index to 1.438. I'd expect to see a colored Becke line
} if the
} dispersion values of ethylene glycol and xtal were close. With a
} little
} more work and a table of two you could predict the color. Cross your
} polars
} (if you haven't done it all ready) and the field is black and the
} xtals are
} also at extinction over 360 degees of stage rotation.
}
} If so we know our material is"
} A: cubic or a glass (look for conchoidal fracture to separate those)
} B: refractive index slightly less then 1.438
} C: EDS found large amounts of Ca and F
}
} Already I'd be thinking about the report I wanted to write.
}
} So why am I taking your time up? Because I see a tendency to look at
} numbers from a computer and accept them as Truth. I've see it in
} technical
} people who say , "If EDS prints out 2 significant figures, then by
} Gawd it
} must be right." I've seen it with people measuring alpha quartz in
} clay
} without regard to peak interference. We need to back check our
} analysis by
} means of a seperate methodology.
}
} Thanks for the rant. Boy, do I miss optical microscopy
}
}
}
} --
} *************************************************************
} Note:
} The information contained in this message may be
} privileged and confidential and protected from disclosure. If
} the reader of this message is not the intended recipient, or
} an employee or agent responsible for delivering this message
} to the intended recipient, you are hereby notified that any
} dissemination, distribution or copying of this communication
} is strictly prohibited. If you have received this
} communication in error, please notify us immediately by
} replying to the message and deleting it from your computer.
} Thank you,
} The Lincoln Electric Company
} **************************************************************
}
}
} ==============================Original
} Headers==============================
} 10, 22 -- From frank_karl-at-lincolnelectric.com Tue Mar 16 10:58:23 2010
} 10, 22 -- Received: from lincolnelectric.com
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} 10, 22 -- In-Reply-To: {201003161506.o2GF6W7p021355-at-ns.microscopy.com}
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} Software
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} }
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} 10, 22 -- X-MIMETrack: CD-MIME by Router on Notescom1/Lincoln
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|_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
Kenneth JT Livi, PhD
The High-Resolution Analytical Electron Microbeam Facility of the
Integrated Imaging Center
Departments of Earth and Planetary Sciences and Biology
Olin Hall
3400 N Charles Street
Johns Hopkins University
Baltimore, Maryland 21218 USA
| | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| |
| :|








==============================Original Headers==============================
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From: klivi-at-jhu.edu
Date: Tue, 16 Mar 2010 11:46:28 -0500
Subject: [Microscopy] Re: Beam Damage

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

Dear Stephane,

I have written an email to the Listserve addressing this a long time
ago. I guess this is a good time to go over it again...

The first thing is that if you are doing TEM, you MUST (and I can't
emphasize this more) determine if you are effecting your sample in any
way. It is what I affectionately refer to as the "Uncertainty
Principle of Microscopy" (apologies to Heisenberg)... in many cases,
the act of observing a sample in the TEM will alter its properties.
This is particularly evident when dealing with valence state
measurements of minerals and hydrous compounds. I have one
particularly difficult material which seems to change its structure
simply by being placed in the vacuum (and this is not just a
dewatering phenomenon).

The most common problem is either ionization or knock-on damage.
Taking a quote from Csencsits and Gronsky (1988) Vitrification of
zeolite Y in the TEM. Zeolites 8, 122-126, "The types of damage
possible in the TEM can be classified under two general headings:
knock-on and radiolytic. “Knock-on” damage involves the interaction of
the incident electron with the nucleus of an atom in the specimen. An
atom is “knocked” from its site, thereby changing the structure.
Radiolytic damage involves the transfer of energy from the incident
electron to the valence electrons in the specimen. The increase in
energy of the specimen electrons results in bond breakage and,
consequently, in the possible alteration of the structure. From my
experience, damage can be very complicated as to the actual mechanism
of damage. In most cases, one can observe something changing during
irradiation such as amorphization of the structure. However, I've had
clues as to hidden things changing that do not seem to effect the
diffraction pattern. It is often the case that transition elements
will change their valence states on the upper and lower sample
surfaces, while the interior is still unchanged. Along with the
valence change comes a loss of oxygen (reduction) and or hydrogen
(oxidation).

Composition plays a part in beam damage. For minerals, those that have
significant Fe in them are more stable than those having say Mg in the
same structure (e.g., pyroxenes and micas). Those with water are less
stable. Also, those with the structures closer to a glass (e.g.,
quartz and framework silicates) are less stable. Li and Na are often
problematic elements in minerals, but possibly not in engineered
materials. Mn can go both ways.

In all materials, defective areas are less stable than well
crystallized regions. So damage will nucleate first at the very defect
you want to observe. One may consider the top and bottom of the sample
as a major defect. this is where damage tends to begin. Sputtering is
common. Also, a metastable phase tends to damage faster than a
thermodynamically stable phase. (Of course, many materials are
thermodynamically unstable in the TEM!)

One can experiment with changing accelerating voltage... sometimes
going higher helps (decreasing the mean-free path in zeolites),
sometime lower helps (e.g. 80 kV for C-nanotubes). Cooling can
decrease element volatilization or migration during EDS analyses. I
have also heard that increasing the temperature can help structural
recovery from beam damage.

The thing to do is "creep up" on your sample. Expose your sample
gently in the beginning as to determine what the initial state of the
material is--then increase the exposure to see if it changes. Keep
your beam spread wide and current down. Unfortunately, this means that
your CCD exposure time will go up and you will have to battle sample
drift. It is usually a compromise between beam damage and noisy CCD
images (or film). Make sure that you order your in scope experiments
in such a way that you reduce damage. I normally get diffraction first
(very gentle), low mag imaging next, then HRTEM, and lastly EDS. This
means that sometimes you don't find out that the stuff you're looking
at is the wrong material until after the EDS analysis. You may have to
sacrifice some material to EDS to determine which particles are the
right ones.

OK. Just a little bit of my experiences with the Uncertainty Principle
of Microscopy.

Ken

On Mar 16, 2010, at 11:11 AM, Stephane Nizet wrote:

} Hi Ken!
}
} I recently attended a workshop from FEI here in Europe and I asked a
} very simple but very hard question to FEI application specialists:
} "What temperature can be reached locally in the TEM?" The answer
} was: probably several thousands of celsius degrees !!
} Indeed I sometimes have the impression that mineral particles melt
} under the electron beam. Unfortunately I am not trained to do E-
} crystallography so I cannot verify if the material becomes amorphous
} or not. Nevertheless, I wanted to ask you how you deal with this
} problem? You are talking about beam damage, it is nice to be aware
} of it, but how do you avoid it? Do you have a cooling stage in your
} TEM? Would it help?
}
} Best regards,
} Stephane
}
}
}
}
}
} ----- Original Message ----
} From: "klivi-at-jhu.edu" {klivi-at-jhu.edu}
} To: nizets2-at-yahoo.com
} Sent: Tue, March 16, 2010 4:02:22 PM
} Subject: [Microscopy] Re: Mineral Identification Software
}
}
}
}
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}
} Dear Melina,
}
} I have to deal with the question you ask all the time in my
} profession. In order to identify a mineral, I rely on three things: 1)
} structure, 2) composition, 3) locality or source. Structure is the
} least ambiguous and the easiest way to identify a mineral is by powder
} X-ray diffraction. However, X-ray identification is made easier by
} knowledge of the composition. In the case where you have very little
} of the sample, single crystal precession X-ray photography can work
} well, but not many people have that technique. The TEM can combine
} diffraction and EDS on small samples, so it is my method of choice.
} However, you have to consume part of the sample to do TEM.
}
} In doing mineral ID, I always use knowledge of the source as a way to
} narrow down the possibilities. I rarely use composition alone to
} positively ID a mineral... especially from EDS data from an SEM. The
} error in these analyses can be quite high and when you take that into
} account, you may not be able to narrow down the possibilities. In
} addition, solid solution of elements within many mineral structures
} makes compositional ID difficult.
}
} If you want to narrow the possibilities down, go to a mineral database
} like: www.webmineral.com (not the only one). There you will find some
} possible candidates. However, you will not find an exact match to your
} composition. With your example, I would first guess fluorite CaF2,
} although the C is problematic. Diffraction information on the TEM
} could easily determine if the material is fluorite. (Watch out for
} beam damage though!)
}
} Mineral ID takes some experience and I don't think there can be a
} quick look up list that will give you an easy answer just from
} composition. To be unequivocal, you need diffraction data.
}
} Hope this helps,
} Ken
}
}
} On Mar 15, 2010, at 7:58 AM, mmiralles-at-pi.ac.ae wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
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} } ----------------------------------------------------------------------------
} }
} } Hi,
} }
} } In relation to mineral identification, is there an available software
} } that can aid mineral identification using the Quant values from the
} } EDS?
} } Let's say, if I have around 58% F, 33% Ca and 7% C.
} }
} } Will appreciate an input.
} }
} } Thanks,
} } Melina
} }
} }
} } ==============================Original
} } Headers==============================
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}
} |_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
} Kenneth JT Livi, PhD
} The High-Resolution Analytical Electron Microbeam Facility of the
} Integrated Imaging Center
} Departments of Earth and Planetary Sciences and Biology
} Olin Hall
} 3400 N Charles Street
} Johns Hopkins University
} Baltimore, Maryland 21218 USA
} | | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| |
} | :|
}
}
}
}
}
}
}
}
} ==============================Original
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}

|_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
Kenneth JT Livi, PhD
The High-Resolution Analytical Electron Microbeam Facility of the
Integrated Imaging Center
Departments of Earth and Planetary Sciences and Biology
Olin Hall
3400 N Charles Street
Johns Hopkins University
Baltimore, Maryland 21218 USA
| | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| |
| :|









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From: baskin-at-bio.umass.edu
Date: Tue, 16 Mar 2010 12:44:23 -0500
Subject: [Microscopy] EM facility director job open

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

Greetings,
This is an exciting job for the right person -- please
ciruclate widely. Thanks. Tobias

Facility Director

The University of Massachusetts Amherst invites applications for a
state-supported position as a Facility director, at the rank of
lecturer, to manage the W. M. Keck Electron Microscopy Center located
in the Conte Center for Polymer Research. The facility director will
operate a university facility which serves users in soft materials,
life sciences, geology, chemistry, physics, and engineering. The
director will manage a suite of instruments for transmission and
scanning electron microscopy, including a newly acquired FEI Magellan
field-emission scanning electron microscope, with low voltage, STEM,
and analytical capability (EBSD and EDS). Duties include assisting
faculty, students, and research staff in instrumentation usage, user
training, facility management, technician supervision, and classroom
instruction/workshops. The director will be research active
(independently and/or collaboratively) and involved in proposal
development. Experience with sample preparation and electron
microscopy aspects of soft (biological and/or polymer) materials is a
plus, and interpersonal skills are needed to interact with the broad
user base. Significant experience using SEM and TEM for research is
required. A Ph.D. in an appropriate materials discipline (Materials
Science, Chemical Engineering, Polymer Science) is required.
Interested individuals should send a letter of interest, a CV, and
the contact information of three references to Dr. Samuel P. Gido,
Search Committee Chair and Associate Professor, Polymer Science and
Engineering Department, University of Massachusetts, Amherst, MA,
01003. The position is expected to be filled by June 2010. Review of
applicants will begin by April 15, 2010, and will continue until the
position is filled. Continuation of position beyond six years is
contingent upon funding. Salary is commensurate with skills and
experience.
The University of Massachusetts is an Equal Opportunity Affirmative
Action Employer; women and members of minority groups are encouraged
to apply.


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From: rintugr-at-gmail.com
Date: Tue, 16 Mar 2010 18:55:38 -0500
Subject: [Microscopy] Core facilities and recent budget cuts

Contents Retrieved from Microscopy Listserver Archives
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Dear Colleagues,

I would like to find out if the university core microscopy facilities
are facing any sort of budget cuts as my university and possibly many
others are trying to close budget gaps due to the recent economic
downturn.

Thank you very much.

Soumitra Ghoshroy


Director, Electron Microscopy Center
Research Associate Professor, Biology
University of South Carolina
715 Sumter Street
Columbia, SC 29208
803-777-7085 (office)
803-777-8908 (fax)
http://www.emc.sc.edu

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From: ldemp-at-mse.ufl.edu
Date: Tue, 16 Mar 2010 22:40:26 -0500
Subject: [Microscopy] viaWWW: Travel Grants for MSA Student members

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This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: ldemp-at-mse.ufl.edu
Name: Amelia Dempere

Organization: Microscopy Society of America

Title-Subject: [Filtered] Travel Grants for MSA Student members

Message: MSA Students members can apply for an MSA Travel Grant to
attend the 2010 IMC17 Congress, to be held in September in Rio de
Janiero. To compete for one of the six $1,000.00 grants available,
students must have an abstract submitted and accepted for a poster or
paper presentation at IMC17. The abstract submission deadline to
IMC17 is April 15,2010 (http://www.imc17.com), and the deadline to
apply for this travel grant is June 30th.
Please contact Dr. Amelia Dempere (ldemp-at-mse.ufl.edu) if you have any
questions. The application form for the Travel Grant can be found at
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From: drg.mitchell-at-sydney.edu.au
Date: Tue, 16 Mar 2010 22:41:11 -0500
Subject: [Microscopy] viaWWW: TEM: EDS on a JEOL 2100 Cryo

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Email: drg.mitchell-at-sydney.edu.au
Name: Dr David RG Mitchell

Organization: ACMM, University of Sydney

Title-Subject: [Filtered] TEM: EDS on a JEOL 2100 Cryo

Message: Dear All

I was wondering if anyone out there has a JEOL 2100 TEM with cryo
polepieces and an EDS system. We are considering this configuration.
However, as there is an additional hole in the anti-contamination
shroud (for the EDS detector), there is the prospect of more ice
formation on cryo specimens, compared with a standard cryo 2100. Our
main focus is on cryo work, but we do want to do some analytical work
also.

Any comments from anyone with direct experience of a cryo 2100/EDS,
would be most welcome.

Thanks in advance and regards, Dave Mitchell, Laboratory Manager,
EMU, University of Sydney.

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From: klivi-at-jhu.edu
Date: Thu, 18 Mar 2010 11:08:21 -0500
Subject: [Microscopy] Used CCD for Philips 420

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I am looking into the possibility of obtaining a used CCD camera for a
Philips 420 TEM... side or bottom mounted. Anyone know of one?

Ken






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From: aaron.cavosie-at-upr.edu
Date: Thu, 18 Mar 2010 11:46:35 -0500
Subject: [Microscopy] SEM: help diagnosing stereoscan 120 electronics failure

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Dear list:

My local SEM is a Cambridge Instruments Stereoscan 120, ca.
~1990-1995. It is equiped only with SE and BSE detectors. Its old, but
it does an okay job.

This week we had a massive failure, very likely resulting in (or
caused by) the failure of an electrical component.

I have a fair amount of microscopy experience as a user/operator of
SEMs (geoscience applications), but am barely a novice with SEM
electronics.

At this point we are trying to troubleshoot what went wrong. If the
problem below sounds familiar to anyone, I would appreciate any
indications of what to give further scrutiny to. (at this point we
have done visual inspection of most of the electronics and have not
noticed anything obviously 'burned-out')

Here is a brief synopsis of the failure:

(1) Normal start-up with electronics and pumping, sample in the
chamber. A slight 'hissing' sound occurred immediately after the rough
pump turned on (does not normally happen), but it went away and the
total pump-down time was identical to normal (~9-10 minutes).

(2) Beam was turned on, SE image seen on screen.

(3) After ~10 seconds, an audible 'bang' or 'pop' occurred, followed
shortly by a burning smell (one person thought it smelled like
rubber...). It seemed to come from the main electronics console,
located below the controls in front of where the operator sits.

(4) Immediately after the 'bang', the entire system died instantly.
Vac, electronics, everything. This is how it is now.

(5) As is, there are no active electronics on the instrument. Nothing
comes on. I tried the rough pump manually by plugging it into a
electrical socket by itself, and it fires up, but everything that runs
off of SEM electronics is dead in the water.

Another interesting observation: when the chamber was opened shortly
after the failure, the same 'burning' smell came from it. This is a
little strange, but there are two possibilities to explain it: (1) the
burning smell originated in the chamber (I don't think so), or (2)
when the chamber vented to atmosphere, the input hose sucked in some
of the ambient burning odor. This may not be an important observation,
but it was noted all the same.

At this point, it is difficult to troubleshoot what caused the
failure, and for that matter, exactly what failed.

It seems to me that the power supply is likely a culprit, given that
no electronics function, but I could easily be off-base.

Any insights to what happened, or suggestions for further diagnosis
would be much appreciated.

Also- if anyone has wiring schematics for the stereoscan 120 and would
be able to provide a copy, I would be most appreciative. I have a
feeling we're going to have to call in some help to get passed this
one.

With best regards,

Aaron

--
************************************************
Aaron Cavosie, Ph.D.
Associate Professor
Department of Geology
P.O. Box 9000
University of Puerto Rico
Recinto Universitario de Mayaguez (RUM)
Mayaguez, PR 00681-9000
Office Phone: (787) 832-4040 ext. 3541
Fax: (787) 265-3845
Email: aaron.cavosie-at-upr.edu
Departmental web page: http://geology.uprm.edu/professors/cavosie.html
Professional web page: http://blogs.uprm.edu/aaroncavosie/
************************************************

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19, 15 -- Subject: SEM: help diagnosing stereoscan 120 electronics failure
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From: gary-at-gaugler.com
Date: Thu, 18 Mar 2010 13:10:09 -0500
Subject: [Microscopy] Re: SEM: help diagnosing stereoscan 120

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Take a very close look at all of the circuit boards and
any modules with electrolytic capacitors. From that
generation, they will typically be cylindrical with a
flat top. Look for any that have a bulging top. Over
time, these capacitors dry out and short. Also, check
all fuses. A blown fuse will likely indicate the source
of the failure. I've seen many boards with multiple
dried out caps. Some will explode. If any in one or
more power supplies have dried out, that will shut down
the system.

While searching for the caps also note if any high wattage
resistors are discolored. These resistors are typically
2W, 5W or 10Watts. They will be part of the bypass circuitry
along with the caps.

If you can get schematics, that would help trace from input
AC to the various DC points of interest. If a three lead
regulator failed, that would be easy to track down.

In any case, it is probably a lot of grief for a $2 part.

gary g.


At 09:47 AM 3/18/2010, you wrote:



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From: chris.millward-at-lineone.net
Date: Fri, 19 Mar 2010 05:33:58 -0500
Subject: [Microscopy] Re: SEM: Help diagnosing stereoscan 120

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Examine all your power supply boards. All large capacitors have a
pressure release point at the top.
In very old capacitors this was a raised dimple which will be obviously
ruptured. The more modern
capacitors have a flat disc that will rupture. The rupture gives a sharp
pop and is often accompanied
by a cloud of white smoke. The circuit board may show obvious signs of
the waxy dielectric in the vicinity.

Although a capacitor has ruptured the circuit board can continue often
functioning. In your case the loss of
power would suggest that the fault lies nearby in some other component.

Chris Millward


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From: fionacxr-at-tll.org.sg
Date: Fri, 19 Mar 2010 11:06:36 -0500
Subject: [Microscopy] viaWWW: Textbooks on EM

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Email: fionacxr-at-tll.org.sg
Name: Fiona Chia

Organization: Temasek Lifesciences Laboratory

Title-Subject: [Filtered] Textbooks on EM

Message: Hi everyone

Does anyone have any suggestion/reccomendations on good textbooks on
SEM and TEM? I'm looking for these and there are many out there and I
am not sure which one to get.

thanks

Regards
Fiona Chia

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From: reznik-at-ict.uni-karlsruhe.de
Date: Fri, 19 Mar 2010 11:07:03 -0500
Subject: [Microscopy] viaWWW: SCD 040_SEM_Coater Problems

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Organization: Karlsruhe University

Title-Subject: [Filtered] SCD 040_SEM_Coater

Message: Hello,

we have a problem with a Balzers SEM-Coater "SCD 040":
there is no current. All fuses are o¥k.
Does anybody know how to solve this problem?

Boris


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From: yu_zongsen-at-yahoo.com
Date: Fri, 19 Mar 2010 11:07:40 -0500
Subject: [Microscopy] Zero-loss peak drifting

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Dear Colleagues


Our newly-installed JEOL 2100F is equipped with Tridiem system. The annoying thing is that Zero-loss peak is always randomly drifting with time, for instance, a few eV within half hour.

I would like to ask you for advice how we can improve that to keep Zero-loss peak more stable (no drifting !).

Thank you !


Leo




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9, 25 -- From: zongsen yu {yu_zongsen-at-yahoo.com}
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From: dac-at-research.umass.edu
Date: Fri, 19 Mar 2010 11:36:47 -0500
Subject: [Microscopy] Re: viaWWW: Textbooks on EM

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

Hi Fiona,

I may not be aware of all the books out there now but if I could have
only one textbook to cover the essentials of the microscope hardware and
vacuum systems and clear, well illustrated, details of specimen
preparation, imaging, interpretation, I would definitely get the Bozzola
and Russell book "Electron Microscopy, Principles and Techniques for
Biologists". It clearly covers the basics and much more. A few copies
are sometimes found on Amazon and eBay. There is an older edition that
still covers the basics very well. Our university has an electronic
version subscription which is another way to get it.

Dale Callaham
Central Microscopy Facility
Univ of Massachusetts -at- Amherst

fionacxr-at-tll.org.sg wrote:
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} Title-Subject: [Filtered] Textbooks on EM
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} Message: Hi everyone
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} Does anyone have any suggestion/reccomendations on good textbooks on
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From: klivi-at-jhu.edu
Date: Fri, 19 Mar 2010 12:47:09 -0500
Subject: [Microscopy] Re: Zero-loss peak drifting

Contents Retrieved from Microscopy Listserver Archives
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Boris:

Did you check out the internal fuses? There are some on the internal power
supply boards.


Al Coritz
--------------------------------------------------
X-from: {reznik-at-ict.uni-karlsruhe.de}
Sent: Friday, March 19, 2010 12:07 PM
To: {Sampleprep-at-earthlink.net}

Dear Leo,
ZLP drift can be due to many factors. It would be a good idea to track
the ZLP over a long period of time and see if the drift correlates
with time of day, happenings in the building, or time after starting
the analyses. We have a much older GIF 200, and we have drift which we
could correlate with: 1) time after switching from image mode to
spectroscopy mode, 2) cooling (on a 90 minute cycle), 3) charging at
the bore at the bottom of the projection chamber. We placed a
dedicated chiller on the GIF, then placed an Al flange in the
projection chamber bore. These helped with drift, but did not
eliminate it. My best data is collected when I can place a sample in
the scope the day before, set up the analysis conditions, switch on
spectroscopy mode and align the GIF. In the morning, the ZLP is more
stable. Not everyone has the luxury of doing this, but when the data
is important, you go the extra mile...
Ken

On Mar 19, 2010, at 12:11 PM, yu_zongsen-at-yahoo.com wrote:

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} Dear Colleagues
}
}
} Our newly-installed JEOL 2100F is equipped with Tridiem system. The
} annoying thing is that Zero-loss peak is always randomly drifting
} with time, for instance, a few eV within half hour.
}
} I would like to ask you for advice how we can improve that to keep
} Zero-loss peak more stable (no drifting !).
}
} Thank you !
}
}
} Leo
}
}
}
}
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} 9, 25 -- From: zongsen yu {yu_zongsen-at-yahoo.com}
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|_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
Kenneth JT Livi, PhD
The High-Resolution Analytical Electron Microbeam Facility of the
Integrated Imaging Center
Departments of Earth and Planetary Sciences and Biology
Olin Hall
3400 N Charles Street
Johns Hopkins University
Baltimore, Maryland 21218 USA
| | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| |
| :|








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From: PhillipsT-at-missouri.edu
Date: Sat, 20 Mar 2010 15:16:08 -0500
Subject: [Microscopy] digital cameras - difference between research and consumer models

Contents Retrieved from Microscopy Listserver Archives
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Hello Boris:
In our SCD040 this is a typical failure. Normally one of the rectifier
diodes on the high voltage power supply fails (1M400X). Most common
cause is trying to do ion etching. We did not attempt to modify the
bridge as it is easy to repair and a kind of fuse.
Hope this helps.
kiss


Francisco José Kiss
Dpt. of Metallurgy.
Federal University of Rio Grande do Sul
Brasil

---------- Original Message -----------
X-from: reznik-at-ict.uni-karlsruhe.de
To: kiss-at-demet.ufrgs.br
Sent: Fri, 19 Mar 2010 11:20:17 -0500

I need to explain to some students the difference between a high end research-grade digital camera and one used for taking photos of friends. Clearly cooling to reduce the dark field current is a factor. The absence of a Bayer filter is another. What I would really like is a table that compared features like spectral sensitivity, gain, dynamic range, S/N, etc for research cameras vs consumer models. Does anyone know of such a table? Thanks, Tom

Thomas E. Phillips, Ph.D.
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
Biological Sciences
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (voice)
573-882-0123 (fax)




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From: dkoleary-at-verizon.net
Date: Sun, 21 Mar 2010 14:47:00 -0500
Subject: [Microscopy] Polarized Light Microscopy Workshop

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New YorkMicroscopical Society
One Prospect Village Plaza
(66F Mt. Prospect Avenue)
Clifton, NJ 07013
Bernard Friedman Memorial Workshop
 
Polarized Light Microscopy
 May 1, 8, 15 & 22, 2010
 
An advanced course on polarized light microscopy which will cover the following topics:
The nature of polarized light
The origin and interpretation of interference colors
Birefringence and crystal orientation,  The Indicatrix
Compensation and variable compensators
Interference figures and their interpretation
 
The workshop will consist of four Consecutive Saturdays of lectures and hands on labs to cover the theoretical and practical aspects of polarized light microscopy. The course instructors include Jan Hinsch formally of Leica, Inc., Mary McCann of McCann Imaging, John Reffner of John Jay College and N.Y.M.S. Instructor Don O'Leary.
 
WHEN: May 1, 8, 15 & 22, 2010 from 10 A.M. to 4 P.M.
 
WHERE: One Prospect Village Plaza. Clifton, NJ Phone (973) 470-8733
 
COST: $425 for N.Y.M.S. members, $455 for non-members (includes membership). Lunch and course materials are included. Checks made out to N.Y.M.S.
 
WHO: advanced course for those who have completed "The Use of the Microscope" or are experienced in microscopy and familiar with the theory of its use.                   
 
HOW: Register using the form below.Limited to the first 12 registrants.
Return form to Don O'Leary, 10 Sampson Street, Unit 113, Saddle Brook, NJ 07663
FURTHER INFORMATION: Call D. O'Leary (201)519-2176 e-mail dkoleary-at-verizon.net
 
                                  PLEASE POST
--------------------------------------------------------------------------
                               Registration Form
                          Polarized Light Microscopy
 
N.Y.M.S. Member_________________ ($425)  Non-Member__________($455)
 
Name_____________________________________________________________
Address___________________________________________________________
Phone (W)_________________________(H)______________________________
e-mail________________________________________


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From: nicholls-at-uic.edu
Date: Mon, 22 Mar 2010 09:15:02 -0500
Subject: [Microscopy] Re: Zero-loss peak drifting

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Leo

For better stability it is important to leave the HT at 200keV overnight
and not to reduce it. The HT tank will cause drift as its temperature
changes after bringing it up and this may occur for periods of up to 6
hours after reaching 200kV. This drift has negligible effect on normal
imaging, only EELS.

If your microscope has a camera chamber for film the prism that puts the
numbers onto film may be charging. This should be given a thicker
conductive coating, or, if film is not being used, remove it.

Finally I assume you do not have steel chairs near the microscope? Wheeling
a steel chair in front of the GIF will cause a shift of several eV. We
have a wooden chair with no wheels that is placed in front of the
microscope to stop this effect!

Alan


At 11:08 AM 3/19/2010, yu_zongsen-at-yahoo.com wrote:



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From: dsoren-at-umich.edu
Date: Mon, 22 Mar 2010 10:07:50 -0500
Subject: [Microscopy] TEM liver-pictures

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

Hello again,

Many thanks to all of you who have responded to my question about
liver fixation for TEM. Some of you asked for more details about
the fixation, dehydration, and infiltration protocol. Others
wanted to see some LM images showing the white holes and canal-like
structures in question.

Here is a link for some LM images of the liver, one showing the canal-
like
structures and the other showing the white holes.

http://www.med.umich.edu/cdb/mil/docs/temimages2.html


The mice whose liver showed the white holes were anesthetized with CO2.
The mice whose liver showed the canal-like structures were anesthetized
with 10 mg/ml Ketamin plus 1 mg/ml Zylazine in PBS.

A cardiac perfusion was done, first by flushing the vasculature with
Sorensens buffer and then perfusing with 3% paraformaldehyde and 2.5 %
glutaraldehyde in 0.1 M Sorensen's buffer, pH 7.4. No anticoagulant
was used in the perfusion. The perfusion lasted for 5-10 minutes,
until the liver was completely stiffened.

A piece of liver was excised and sliced under fixative into 1 X 1 X2 mm
pieces. Then it was immersion fixed for two days at 4 degrees.

The tissue was processed further in a Leica EM tissue processor. It was
post fixed in 1 % osmium tetroxide in 0.1 M Sorensens buffer for 1 hour,
rinsed in the same buffer and then dehydrated. It went through 30,
50, 70,
95, and two changes of 100 % EtOH for 20 minutes each. Next, it went
through 3 changes of propylene oxide for 20 minutes each. It was
infiltrated for 4 hours in 3:1 PO:Epon, 6 hours in 1:1 PO:Epon, and 8
hours
in 1:3 PO:Epon. Then it went through two changes of full
strength resin, one for 6 hours and the other one overnight. The full
strength resin infiltration was done in a vacuum desiccator to help
draw off residual PO. Lastly, the samples were transferred into block
with new resin and
polymerized for 24 hours at 60 degrees. The tissue was not en bloc
stained.

Some of the comments from my first posting included some very good
suggestions, such as cutting smaller pieces of tissue before immersion
fixation, using an anti-coagulant with the perfusion, inadequate
infiltration of resin, the white holes could be areas of extracted
lipid or
extracted glycogen. Based on this additional information, can anyone
offer
any further insight as to what might be causing these structures?

As always, thank you so much for your suggestions.

Dotty


Original post:

Hello all,

Does anyone have a tried and true protocol for processing liver for
TEM? We seem to be having problems with poor fixation. We have been
perfusing with 3% p.f. and 2.5 % glutaraldehyde in Sorensen's buffer,
followed by immersion fixation in the same mixture. We post fix with
1 % osmium in the same buffer, dehydrate with a graded series of
ethanol, change to propylene oxide, and then infiltrate and embed in
Epon. The cells immediately adjacent to the blood vessels and right
on the cut edge of the tissue are well fixed, but the cells located
on the interior of the tissue exhibit little white holes. The tissue
had been cut to about 1x2x2 mm. Another problem that we have
encountered in the past is cleared areas that remind me of canals
running through the cytoplasm. Is this to be expected? We would
love to hear from advice from some liver experts out there.

Thanks,

Dotty




Dorothy Sorenson
Microscopy and Image-analysis Laboratory
Department of Cell and Developmental Biology
University Of Michigan Medical School
A807 BSRB
109 Zina Pitcher Place
Ann Arbor, MI 48109-2200
(734)763-1170
FAX (734)763-1166



==============================Original Headers==============================
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From: PhillipsT-at-missouri.edu
Date: Mon, 22 Mar 2010 10:14:50 -0500
Subject: [Microscopy] TEM liver-pictures

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

I agree that it is easy to extract lipid from tissue during processing but disagree that glycogen is easily extracted. It is easily left unstained which makes large deposits look empty. But the glycogen usually remains in place. The holes don't look like glycogen deposits to me but if you wanted to check an appropriate carbohydrate stain such as Thiery's method would reveal it. PAS can also work on etched LM sections.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: dsoren-at-umich.edu [mailto:dsoren-at-umich.edu]
Sent: Monday, March 22, 2010 10:08 AM
To: Phillips, Thomas E.

Hello again,

Many thanks to all of you who have responded to my question about
liver fixation for TEM. Some of you asked for more details about
the fixation, dehydration, and infiltration protocol. Others
wanted to see some LM images showing the white holes and canal-like
structures in question.

Here is a link for some LM images of the liver, one showing the canal-
like
structures and the other showing the white holes.

http://www.med.umich.edu/cdb/mil/docs/temimages2.html


The mice whose liver showed the white holes were anesthetized with CO2.
The mice whose liver showed the canal-like structures were anesthetized
with 10 mg/ml Ketamin plus 1 mg/ml Zylazine in PBS.

A cardiac perfusion was done, first by flushing the vasculature with
Sorensens buffer and then perfusing with 3% paraformaldehyde and 2.5 %
glutaraldehyde in 0.1 M Sorensen's buffer, pH 7.4. No anticoagulant
was used in the perfusion. The perfusion lasted for 5-10 minutes,
until the liver was completely stiffened.

A piece of liver was excised and sliced under fixative into 1 X 1 X2 mm
pieces. Then it was immersion fixed for two days at 4 degrees.

The tissue was processed further in a Leica EM tissue processor. It was
post fixed in 1 % osmium tetroxide in 0.1 M Sorensens buffer for 1 hour,
rinsed in the same buffer and then dehydrated. It went through 30,
50, 70,
95, and two changes of 100 % EtOH for 20 minutes each. Next, it went
through 3 changes of propylene oxide for 20 minutes each. It was
infiltrated for 4 hours in 3:1 PO:Epon, 6 hours in 1:1 PO:Epon, and 8
hours
in 1:3 PO:Epon. Then it went through two changes of full
strength resin, one for 6 hours and the other one overnight. The full
strength resin infiltration was done in a vacuum desiccator to help
draw off residual PO. Lastly, the samples were transferred into block
with new resin and
polymerized for 24 hours at 60 degrees. The tissue was not en bloc
stained.

Some of the comments from my first posting included some very good
suggestions, such as cutting smaller pieces of tissue before immersion
fixation, using an anti-coagulant with the perfusion, inadequate
infiltration of resin, the white holes could be areas of extracted
lipid or
extracted glycogen. Based on this additional information, can anyone
offer
any further insight as to what might be causing these structures?

As always, thank you so much for your suggestions.

Dotty


Original post:

Hello all,

Does anyone have a tried and true protocol for processing liver for
TEM? We seem to be having problems with poor fixation. We have been
perfusing with 3% p.f. and 2.5 % glutaraldehyde in Sorensen's buffer,
followed by immersion fixation in the same mixture. We post fix with
1 % osmium in the same buffer, dehydrate with a graded series of
ethanol, change to propylene oxide, and then infiltrate and embed in
Epon. The cells immediately adjacent to the blood vessels and right
on the cut edge of the tissue are well fixed, but the cells located
on the interior of the tissue exhibit little white holes. The tissue
had been cut to about 1x2x2 mm. Another problem that we have
encountered in the past is cleared areas that remind me of canals
running through the cytoplasm. Is this to be expected? We would
love to hear from advice from some liver experts out there.

Thanks,

Dotty




Dorothy Sorenson
Microscopy and Image-analysis Laboratory
Department of Cell and Developmental Biology
University Of Michigan Medical School
A807 BSRB
109 Zina Pitcher Place
Ann Arbor, MI 48109-2200
(734)763-1170
FAX (734)763-1166



==============================Original Headers==============================
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From: george-gogo-at-live.com
Date: Mon, 22 Mar 2010 11:47:14 -0500
Subject: [Microscopy] viaWWW: Computer for Image Processing

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Email: george-gogo-at-live.com
Name: George

Organization: TU/e

Title-Subject: [Filtered] Computer for Image Processing

Message: Dear All,

I am going to buy a computer for electron tomography, which can use
GPUs for image processing.
my question is whether anyone has any suggestions/recommendations on
the computer configurations.

thank you.

Kind regards,

George

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From: ejb1176-at-gmail.com
Date: Mon, 22 Mar 2010 11:47:43 -0500
Subject: [Microscopy] viaWWW: Seminar - Current Trends in Electron Microscopy and

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Email: ejb1176-at-gmail.com
Name: Edward Basgall

Organization: Drexel University

Title-Subject: [Filtered] Seminar - Current Trends in Electron
Microscopy and Microanalysis - Drexel University, April 14, 2010

Message: Seminar - Current Trends in Electron Microscopy and
Microanalysis - Drexel University, April 14, 2010

You are cordially invited to attend the "Current Trends in Electron
Microscopy and Microanalysis" seminar at Drexel University, April 14,
2010.

Complimentary coffee and lunch will be provided. Free parking will be
available to pre-registered guests.

Please navigate to:
http://crf.coe.drexel.edu/crf/workshops
to register on the CRF website by March 31. Space is limited.

For more information please contact Craig Johnson at (215) 895-5900
or cljohnson-at-coe.drexel.edu



Edward J. Basgall, Ph.D.
Microscopy Manager
Drexel University - Central Research Facility
106 Bossone Research Center
31st and Market Streets
Philadelphia, PA 19104
office: (215) 895-2379


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From: vladislav_speransky-at-nih.gov
Date: Mon, 22 Mar 2010 12:03:13 -0500
Subject: [Microscopy] Re: viaWWW: Computer for Image Processing

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

Dear George Gogo from TU/e,

The only thing I can tell you right away from what you wrote, make
sure you are getting an Nvidia GPU on board (not ATI). Note that
Quadro Nvidia card, on the other hand, is unnecessary for this
purpose. Get a Quadro *only* if you plan to use 3D viewing. (If you
are not sure, then you most likely won't. *Some* people find it
useful, but most will say it is a white elephant.)

Other than that, it is probably wise at this point to get more
processor cores as opposed to fewer cores but higher clock speed. A
couple of years ago, the opposite were true, but now most image
processing packages seem to support multiple cores well (as do
Photoshop and Quicktime engines).

Faster HD, if there is a choice (e.g., 10K vs 7.2K), will let large
volumes load somewhat quicker... but won't do anything for
reconstruction speed, since it is not a limiting factor. And don't go
into that RAID stuff unless you really know what you need.

You are not saying anything about the platform - Mac, Windoze, Linux?
If you are not sure, then definitely not Linux... Otherwise, either
choice is fine.

Vlad
________________________________________________
Vlad Speransky, Staff Scientist
Supramolecular Structure and Function Resource
National Institute of Biomedical Imaging and Bioengineering, NIH
13 South Dr, Rm. 3N17 MSC 5766
Bethesda, MD 20892
301 496-3989
vladislav_speransky-at-nih.gov
http://www.nibib.nih.gov/Research/Intramural/lbps/staff/speransky



[Microscopy] viaWWW: Computer for Image Processing
george-gogo-at-live.com [george-gogo-at-live.com]
Sent:
Monday, March 22, 2010 12:48 PM
To:
Speransky, Vlad (NIH/NIBIB) [E]



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Email: george-gogo-at-live.com
Name: George

Organization: TU/e

Title-Subject: [Filtered] Computer for Image Processing

Message: Dear All,

I am going to buy a computer for electron tomography, which can use
GPUs for image processing.
my question is whether anyone has any suggestions/recommendations on
the computer configurations.

thank you.

Kind regards,

George

Login Host: 131.155.81.102
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23, 23 -- From vladislav_speransky-at-nih.gov Mon Mar 22 12:03:12 2010
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From: nizets2-at-yahoo.com
Date: Tue, 23 Mar 2010 06:42:22 -0500
Subject: [Microscopy] TEM liver-pictures

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

Actually, PAS worked for me on EPON semi-thin sections.
The periodic acid (would) work as an etchant.

30 min at 50°C in 5% periodic acid
30 min at 50°C in Schiff reagent

I post-stained with Azur II at RT for 20 min

Regards,
Stephane


----- Original Message ----
X-from: "PhillipsT-at-missouri.edu" {PhillipsT-at-missouri.edu}
To: nizets2-at-yahoo.com
Sent: Mon, March 22, 2010 4:17:31 PM

I agree that it is easy to extract lipid from tissue during processing but disagree that glycogen is easily extracted. It is easily left unstained which makes large deposits look empty.  But the glycogen usually remains in place. The holes don't look like glycogen deposits to me but if you wanted to check an appropriate carbohydrate stain such as Thiery's method would reveal it. PAS can also work on etched LM sections.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: dsoren-at-umich.edu [mailto:dsoren-at-umich.edu]
Sent: Monday, March 22, 2010 10:08 AM
To: Phillips, Thomas E.

Hello again,

Many thanks to all of you who have responded to my question about
liver fixation for TEM.  Some of you asked for more details about
the fixation, dehydration, and infiltration protocol.  Others
wanted to see some LM images  showing the white holes and canal-like
structures in question.

Here is a link for some LM images of the liver, one showing the canal-
like
structures and the other showing the white holes.

http://www.med.umich.edu/cdb/mil/docs/temimages2.html


The mice whose liver showed the white holes were anesthetized with CO2.
The mice whose liver showed the canal-like structures were anesthetized
with 10 mg/ml Ketamin plus 1 mg/ml Zylazine in PBS.

A cardiac perfusion was done, first by flushing the vasculature with
Sorensens buffer and then perfusing with 3% paraformaldehyde and 2.5 %
glutaraldehyde in 0.1 M Sorensen's buffer, pH 7.4.  No anticoagulant
was used in the perfusion.  The perfusion lasted for 5-10 minutes,
until the liver was completely stiffened.

A piece of liver was excised and sliced under fixative into 1 X 1 X2 mm
pieces.  Then it was immersion fixed for two days at 4 degrees.

The tissue was processed further in a Leica EM tissue processor.  It was
post fixed in 1 % osmium tetroxide in 0.1 M Sorensens buffer for 1 hour,
rinsed in the same buffer and then dehydrated.  It went through 30, 
50, 70,
95, and two changes of 100 % EtOH for 20 minutes each.  Next, it went
through 3 changes of propylene oxide for 20 minutes each.  It was
infiltrated for 4 hours in 3:1 PO:Epon, 6 hours in 1:1 PO:Epon, and 8 
hours
in 1:3 PO:Epon.  Then it went through two changes of full
strength resin, one for 6 hours and the other one overnight.  The full
strength resin infiltration was done in a vacuum desiccator to help
draw off residual PO. Lastly, the samples were transferred into block 
with new resin and
polymerized for 24 hours at 60 degrees.  The tissue was not en bloc 
stained.

Some of the comments from my first posting included some very good
suggestions, such as cutting smaller pieces of tissue before immersion
fixation, using an anti-coagulant with the perfusion, inadequate
infiltration of resin, the white holes could be areas of extracted 
lipid or
extracted glycogen. Based on this additional information, can anyone 
offer
any further insight as to what might be causing these structures?

As always, thank you so much for your suggestions.

Dotty


Original post:

Hello all,

Does anyone have a tried and true protocol for processing liver for
TEM?  We seem to be having problems with poor fixation.  We have been
perfusing with 3% p.f. and 2.5 % glutaraldehyde in Sorensen's buffer,
followed by immersion fixation in the same mixture.  We post fix with
1 % osmium in the same buffer, dehydrate with a graded series of
ethanol, change to propylene oxide, and then infiltrate and embed in
Epon.  The cells immediately adjacent to the blood vessels and right
on the cut edge of the tissue are well fixed, but the cells located
on the interior of the tissue exhibit little white holes.  The tissue
had been cut to about 1x2x2 mm.  Another problem that we have
encountered in the past is cleared areas that remind me of canals
running through the cytoplasm.  Is this to be expected?  We would
love to hear from advice from some liver experts out there.

Thanks,

Dotty




Dorothy Sorenson
Microscopy and Image-analysis Laboratory
Department of Cell and Developmental Biology
University Of Michigan Medical School
A807 BSRB
109 Zina Pitcher Place
Ann Arbor, MI  48109-2200
(734)763-1170
FAX (734)763-1166



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From: marienti-at-tiscali.it
Date: Tue, 23 Mar 2010 09:15:25 -0500
Subject: [Microscopy] SEM ETEC diagrams

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

Hi Listers,
I am based in Milano (Italy) and I am trying to solve a problem on a "vintage" ETEC AUTOSCAN.
+24V is missing on the magnification control drawer despite it is present on the filter capacitor.
There are thyristors in between and I need to understand how the circuit work.
Anybody out there can provide at least the diagram of the power supply? (better the complete diagrams).

Marco

Marco Arienti
www.electronrescue.com
mobile +39-335-5619301

==============================Original Headers==============================
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From: kenconverse-at-qualityimages.biz
Date: Tue, 23 Mar 2010 11:47:18 -0500
Subject: [Microscopy] SEM ETEC diagrams

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

Marco,
Send me a mailing address and I'll send you a DVD with schematics, User's
and Service manuals and a diagnostic guide.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: marienti-at-tiscali.it [mailto:marienti-at-tiscali.it]
Sent: Tuesday, March 23, 2010 10:18 AM
To: kenconverse-at-qualityimages.biz

Hi Listers,
I am based in Milano (Italy) and I am trying to solve a problem on a
"vintage" ETEC AUTOSCAN.
+24V is missing on the magnification control drawer despite it is present on
the filter capacitor.
There are thyristors in between and I need to understand how the circuit
work.
Anybody out there can provide at least the diagram of the power supply?
(better the complete diagrams).

Marco

Marco Arienti
www.electronrescue.com
mobile +39-335-5619301

==============================Original Headers==============================
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From: sergei2-at-ornl.gov
Date: Tue, 23 Mar 2010 14:20:17 -0500
Subject: [Microscopy] SPM for Energy Applications

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

Dear colleagues

The Center for Nanophase Materials Sciences at Oak Ridge National Laboratory (ORNL) and Asylum Research Corporation are co-organizing the International Workshop for Scanning Probe Microscopy for Energy Applications, to be held at ORNL September 15-17, 2010. This workshop of invited and contributed talks will cover the recent advances in characterization of energy-relevant materials systems using SPM/AFM techniques, as well as the state of the art in energy dissipation and transformation measurements by SPM/AFM. The three-day meeting will also include a poster session, as well as an equipment lab and hands-on tutorials for demonstration of recently developed dynamic and multi-spectral SPM/AFM modes. The keynote talk will be on “Local Probing of Carrier Dynamics in Polymer Photovoltaic Materials” by David Ginger of the University of Washington.
Major topics to be covered include:
• Mapping of carrier dynamics and photoinduced behavior of photovoltaic materials
• Ionic and electronic transport in fuel cells and Li-ion batteries
• Energy harvesting by piezoelectric and ferroelectric systems,
• Novel advances in functional probes – microwave, thermal, and conductive
• Imaging energy transformations and dissipation by multimodal and Band Excitation SPM/AFM
Detailed information on the agenda, presentations, and registration can be found at www.asylumresearch.com/Energy

Yours

Sergei Kalinin

--
Sergei V. Kalinin
co-Theme Leader for Functional Imaging on the Nanoscale
The Center for Nanophase Materials Sciences
and Materials Sciences and Technology Division
Oak Ridge National Laboratory
Oak Ridge, TN 37922

Section Editor, Nanotechnology

Adjunct Associate Professor,
Department of Materials Science and Engineering,
University of Tennessee, Knoxville

Phone: (865) 241-0236
http://imaging.ornl.gov


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9, 24 -- From: "Sergei V. Kalinin" {sergei2-at-ornl.gov}
9, 24 -- Subject: SPM for Energy Applications
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From: Elliott-at-arizona.edu
Date: Tue, 23 Mar 2010 17:03:40 -0500
Subject: [Microscopy] Re: viaWWW: Textbooks on EM

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

I second the suggestion of Bozzola and Russell. Here are a few others
that I like.
Hunter "Practical Electron Microscopy" - mostly beginner stuff, but
great for beginners.
Maunsbach & Afzelius "Biomedical Electron Microscopy" - this is the
first book I look to for answers and suggestions.
Crang & Klomparens "Artifacts in Biological Electron Microscopy" -
self-explanatory.

David


On Mar 19, 2010, at 9:11 AM, fionacxr-at-tll.org.sg wrote:

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} Title-Subject: [Filtered] Textbooks on EM
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} Does anyone have any suggestion/reccomendations on good textbooks on
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From: wesaia-at-iastate.edu
Date: Tue, 23 Mar 2010 17:23:57 -0500
Subject: [Microscopy] JEOL 840A available

Contents Retrieved from Microscopy Listserver Archives
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The Materials Analysis and Research Lab at Iowa State University is getting ready to phase out its JEOL 840A SEM. The scope has served us well from 1987 up to the present. It is now time to replace it with a new field-emission scope coming in May. We need to move it out at the end of April to make room for the new SEM.

The scope is in good condition and has been under service contract its entire life. Images of the scope are available on our website.
http://www.mse.iastate.edu/cmac/jeol-840a-scanning-electron-microscope.html
Note that the EDS system is NOT included. We will need to use it with the new scope for the time being.

Selected images from the scope are also available on-line.
ftp://www.marl.iastate.edu/General/JEOL_840A/

The scope is basically available free for the taking. We would be happy to take some money for it if someone offered, but our prime concern is getting it out of the way for the new scope. We may be able to help some, but the new owner would be responsible for packing it up and moving it out.

You may contact Jerry Amenson or myself for more details.

Warren Straszheim
wesaia-at-iastate.edu
515-294-8187

Jerry Amenson
jamenson-at-iastate.edu
515-294-8752



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From: nizets2-at-yahoo.com
Date: Wed, 24 Mar 2010 04:14:35 -0500
Subject: [Microscopy] help with diagnostic

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

Dear all,

I am having trouble with our Tecnai G20/SF6. I start the microscope and all is fine, it is ready for action.
I start the HT and I let stabilize for 1 hour. I control the chillerwater temperature and it is stable (17-18°C).
Really all looks perfectly normal.
Then I leave the microscope to go and eat at noon and when I come back the display is frozen, nothing works and the microscope is OFF (the main lamp is on OFF).
I start the PC and the microscope and again everything is fine. I leave it for 2 hours, until I go home and all looks fine.

I come back the next day in the morning the display on the monitor is frozen and the microscope is OFF!!!
I started it again and all looks fine until now, I am controlling everything regularly but in the meantime I thought I would send a message in the bottle to the list to know what you think about this story.

Now some side notes:

- The emission chamber has been filled with SF6 on monday up to 6 bar. I had the time (fortunately) to condition the HT just before the error happened. Actually I went all up to 220kV then I left the microscope for the noon pause on 220kV and when I came back all was frozen (microscope, vacuum and HT off).

- There was a recent change in the water chiller: the water was always set to 15°C in charge and the temperature limit (backflow) was set on 24°C. 2 weeks ago the machine stopped working. When I investigated, I set the temperature limit to 30°C and it started again, the water temperature reaching 19°C in charge (without changing the temperature setting). I let it go because these values are OK I think.

- On the "frozen display", there is not a single error message, nothing. It is just as it was when I left it.

- I controlled the log file and, although I understand almost nothing from the hundreds of messages, it seems that no error occured. There are just info messages, no error messages.

- One strange thing: when the vacuum in the column is broken, there is a red LED lighting on the goniometer. When the problem happens and the display is frozen and the microscope is OFF, the red LED is not lighting. However when I start the system new all the vacuum are broken. I don't understand how the red LED was not lit while the vacuum was broken.

I have 2 hypotheses: maybe it is the chiller, but I wonder how it can work fine for 3 hours and then suddenly fail, then work again and so on...
Or it has to do with the PC, which controlls everything. Something in the PC is unstable and crashes and this crashes the whole system. Unfortunately I know too few in computers to investigate this issue myself.

I would be glad if you'd care to share your thoughts about this issue with me.

regards,

Stephane 






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17, 27 -- From: Stephane Nizet {nizets2-at-yahoo.com}
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From: kenconverse-at-qualityimages.biz
Date: Wed, 24 Mar 2010 06:13:39 -0500
Subject: [Microscopy] help with diagnostic

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

This has got to be the best example I've seen for why you don't want to get
rid of your older, non-PC controlled SEM or TEM. You are not only at the
complete mercy of the manufacturer, you're at the complete mercy of Windows!
Good luck.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Wednesday, March 24, 2010 5:18 AM
To: kenconverse-at-qualityimages.biz

Dear all,

I am having trouble with our Tecnai G20/SF6. I start the microscope and all
is fine, it is ready for action.
I start the HT and I let stabilize for 1 hour. I control the chillerwater
temperature and it is stable (17-18°C).
Really all looks perfectly normal.
Then I leave the microscope to go and eat at noon and when I come back the
display is frozen, nothing works and the microscope is OFF (the main lamp is
on OFF).
I start the PC and the microscope and again everything is fine. I leave it
for 2 hours, until I go home and all looks fine.

I come back the next day in the morning the display on the monitor is frozen
and the microscope is OFF!!!
I started it again and all looks fine until now, I am controlling everything
regularly but in the meantime I thought I would send a message in the bottle
to the list to know what you think about this story.

Now some side notes:

- The emission chamber has been filled with SF6 on monday up to 6 bar. I had
the time (fortunately) to condition the HT just before the error happened.
Actually I went all up to 220kV then I left the microscope for the noon
pause on 220kV and when I came back all was frozen (microscope, vacuum and
HT off).

- There was a recent change in the water chiller: the water was always set
to 15°C in charge and the temperature limit (backflow) was set on 24°C. 2
weeks ago the machine stopped working. When I investigated, I set the
temperature limit to 30°C and it started again, the water temperature
reaching 19°C in charge (without changing the temperature setting). I let it
go because these values are OK I think.

- On the "frozen display", there is not a single error message, nothing. It
is just as it was when I left it.

- I controlled the log file and, although I understand almost nothing from
the hundreds of messages, it seems that no error occured. There are just
info messages, no error messages.

- One strange thing: when the vacuum in the column is broken, there is a red
LED lighting on the goniometer. When the problem happens and the display is
frozen and the microscope is OFF, the red LED is not lighting. However when
I start the system new all the vacuum are broken. I don't understand how the
red LED was not lit while the vacuum was broken.

I have 2 hypotheses: maybe it is the chiller, but I wonder how it can work
fine for 3 hours and then suddenly fail, then work again and so on...
Or it has to do with the PC, which controlls everything. Something in the PC
is unstable and crashes and this crashes the whole system. Unfortunately I
know too few in computers to investigate this issue myself.

I would be glad if you'd care to share your thoughts about this issue with
me.

regards,

Stephane 






==============================Original Headers==============================
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From: mcauliff-at-umdnj.edu
Date: Wed, 24 Mar 2010 08:39:43 -0500
Subject: [Microscopy] EM texts

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

I like "A beginner's handbook in biological transmission electron
microscopy", 2nd edition, by Brenda S. Weakley.

Geoff

--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff-at-umdnj.edu
**********************************************



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From: protrain-at-emcourses.com
Date: Wed, 24 Mar 2010 13:08:45 -0500
Subject: [Microscopy] help with diagnostic

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

Hi Stephane

I am not familiar with the FEI instrument that you have but I am familiar
with the problems that are typical of a modern computer controlled
instruments. What you describe is pretty typical of computer controlled
instruments and without expert knowledge is impossible to cure though
emails.

A service technician, unless he/she has seen the problem before, will start
by checking out the control PC. There is no point crashing in to the
vacuum, lens, or high voltage circuits until you are 100% that it is not
Windows or the base computer that is causing the problem.

If you have not already done so I would carry out the following which take
into account that the control device is simply a PC so you need to carry out
all the tidy up procedures that you should routinely carry out on your desk
top PC.

1. Go to the C drive - Accessories - System Tools - Disk Cleanup - to
remove all the rubbish that collects on the drive.

2. Go to Computer from the Start icon - right click on the C drive -
Properties - Tools - Error checking - Check now - Automatically fix file
system errors AND Scan and attempt recovery of bad sectors - start - and you
will need to let the system close down and re start for the process to be
carried out.

3. Use a well know Registry Fix programme like Registry Fix ( {$40 on
the web) to clean up the registry from all the junk it gathers.

4. Now see how your microscope behaves

5. Still a problem then contact the supplier.

Hope this helps?


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: 24 March 2010 09:16
To: protrain-at-emcourses.com

Dear all,

I am having trouble with our Tecnai G20/SF6. I start the microscope and all
is fine, it is ready for action.
I start the HT and I let stabilize for 1 hour. I control the chillerwater
temperature and it is stable (17-18°C).
Really all looks perfectly normal.
Then I leave the microscope to go and eat at noon and when I come back the
display is frozen, nothing works and the microscope is OFF (the main lamp is
on OFF).
I start the PC and the microscope and again everything is fine. I leave it
for 2 hours, until I go home and all looks fine.

I come back the next day in the morning the display on the monitor is frozen
and the microscope is OFF!!!
I started it again and all looks fine until now, I am controlling everything
regularly but in the meantime I thought I would send a message in the bottle
to the list to know what you think about this story.

Now some side notes:

- The emission chamber has been filled with SF6 on monday up to 6 bar. I had
the time (fortunately) to condition the HT just before the error happened.
Actually I went all up to 220kV then I left the microscope for the noon
pause on 220kV and when I came back all was frozen (microscope, vacuum and
HT off).

- There was a recent change in the water chiller: the water was always set
to 15°C in charge and the temperature limit (backflow) was set on 24°C. 2
weeks ago the machine stopped working. When I investigated, I set the
temperature limit to 30°C and it started again, the water temperature
reaching 19°C in charge (without changing the temperature setting). I let it
go because these values are OK I think.

- On the "frozen display", there is not a single error message, nothing. It
is just as it was when I left it.

- I controlled the log file and, although I understand almost nothing from
the hundreds of messages, it seems that no error occured. There are just
info messages, no error messages.

- One strange thing: when the vacuum in the column is broken, there is a red
LED lighting on the goniometer. When the problem happens and the display is
frozen and the microscope is OFF, the red LED is not lighting. However when
I start the system new all the vacuum are broken. I don't understand how the
red LED was not lit while the vacuum was broken.

I have 2 hypotheses: maybe it is the chiller, but I wonder how it can work
fine for 3 hours and then suddenly fail, then work again and so on...
Or it has to do with the PC, which controlls everything. Something in the PC
is unstable and crashes and this crashes the whole system. Unfortunately I
know too few in computers to investigate this issue myself.

I would be glad if you'd care to share your thoughts about this issue with
me.

regards,

Stephane 






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From: wesaia-at-iastate.edu
Date: Wed, 24 Mar 2010 14:56:17 -0500
Subject: [Microscopy] JEOL 840A available - update

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

The JEOL 840A that I mentioned yesterday is still available. However, as things often go, there has been a glitch.

It was an oversight on my part for not first inquiring with our surplus office regarding laboratory equipment removal. They need to make it public knowledge, through a bid process, that the JEOL 840A is for sale. The bid process must be followed before the instrument can be made available to anyone. They will send out for bids next week, with two weeks allowed to reply and 2-4 weeks to remove the instrument.

We will forward a bid notice to those that have already replied. Again, feel free to contact Jerry or me if you have any questions regarding the instrument.

Regards,
Warren Straszheim
wesaia-at-iastate.edu
515-294-8187

Jerry Amenson
jamenson-at-iastate.edu
515-294-8752


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From: SWu-at-hei.org
Date: Wed, 24 Mar 2010 18:00:21 -0500
Subject: [Microscopy] TEM - Give Away: Film Developing Supplies

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

Due to our recent switch to digital image capture system on our FEI TEM,
we have film developing supplies that are now not needed in our lab.

We would like to give them away FOR FREE; however, you WILL BE
RESPONSIBLE FOR ALL NECESSARY SHIPPING COST.

If you're located near our lab, arrangements can also be made for you to
come and pick up the items yourself.

Items are available on first-come, first-serve basis until April 30,
2010.

For arrangements or questions, please contact me by email to swu-at-hei.org


Bellow is the list of what're available:

=Kodak D-19 Developer
* Kodak catalog# 1464593
* Quantity: 17 pouches, each pouch makes one gallon of working solution.
* Condition: new and unopened

=Kodak Rapid Fixer
* Kodak catalog# 1464106
* Quantity: 6 kits, each kit makes one gallon of working solution.
* Condition: new and unopened.

=Film Racks: EMS Lucite Racks for 3-1/4" x 4" films
* EMS catalog# 47894-25, holds 25 films per rack
* Quantity: 3
* Condition: used but in nice & clean condition

=Film Racks: Ted Pella Lucite Racks for 3-1/4"x4" films
* Ted Pella catalog# 22706, holds 25 films per rack
* Quantity: 3
* Condition: used but in nice & clean condition

=Film Racks: EMS Lucite Racks for 4"x5" films
* EMS catalog# 74895-12, holds 12 films per rack
* Quantity: 2
* Condition: used but in nice & clean condition

=Developing Tanks with Floating Lids (plastic)
* Ted Pella catalog# 26815, fits both 3-1/4"x4"- & 4"x5" racks mentioned
above.
* Quantity: 3
* Condition: used, precipitates from developing chemicals.

=Developing Tanks without Lids (Kodak 5x7 Hard Rubber Tank)
* EMS catalog# 74345, fits both 3-1/4"x4" - & 4"x5"- racks mentioned
above.
* Quantity: 2
* Condition: used, water stain

=Film Holders for FEI BioTwin CM120 TEM
* Holds 56 3-1/4"x4" films on designated magazines
* Quantity: 2
* Condition: practically new

=Film Magazines for 3-1/4"x4" sheet films
* Works with FEI BioTwin CM120 TEM Film Holder mentioned above
* Quantity: 73
* Condition: practically new


Thank you very much,

Siva Wu
Research Assistant
Dept. of Advanced Electron Microscopy & Imaging
Div. of Cell Biology & Genetics
House Ear Institute
2100 West 3rd Street
Los Angeles, CA 90057
(213)989-6754
swu-at-hei.org


==============================Original Headers==============================
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20, 20 -- Subject: TEM - Give Away: Film Developing Supplies
20, 20 -- Date: Wed, 24 Mar 2010 16:00:17 -0700
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20, 20 -- From: "Wu, Siva" {SWu-at-hei.org}
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From: SWu-at-hei.org
Date: Wed, 24 Mar 2010 20:17:32 -0500
Subject: [Microscopy] TEM - Give Away: Film Developing Supplies

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

The items went really quickly.

The only ones left now are these:

=Film Racks: EMS Lucite Racks for 4"x5" films
* EMS catalog# 74895-12, holds 12 films per rack
* Quantity: 2
* Condition: used but in nice & clean condition

=Developing Tanks with Floating Lids (plastic)
* Ted Pella catalog# 26815, fits both 3-1/4"x4"- & 4"x5" racks mentioned
above.
* Quantity: 3
* Condition: used, precipitates from developing chemicals.

=Developing Tanks without Lids (Kodak 5x7 Hard Rubber Tank)
* EMS catalog# 74345, fits both 3-1/4"x4" - & 4"x5"- racks mentioned
above.
* Quantity: 2
* Condition: used, water stain


Siva


-----Original Message-----
X-from: Wu, Siva
Sent: Wednesday, March 24, 2010 4:10 PM
To: Wu, Siva

Due to our recent switch to digital image capture system on our FEI TEM,
we have film developing supplies that are now not needed in our lab.

We would like to give them away FOR FREE; however, you WILL BE
RESPONSIBLE FOR ALL NECESSARY SHIPPING COST.

If you're located near our lab, arrangements can also be made for you to
come and pick up the items yourself.

Items are available on first-come, first-serve basis until April 30,
2010.

For arrangements or questions, please contact me by email to swu-at-hei.org


Bellow is the list of what're available:

=Kodak D-19 Developer
* Kodak catalog# 1464593
* Quantity: 17 pouches, each pouch makes one gallon of working solution.
* Condition: new and unopened

=Kodak Rapid Fixer
* Kodak catalog# 1464106
* Quantity: 6 kits, each kit makes one gallon of working solution.
* Condition: new and unopened.

=Film Racks: EMS Lucite Racks for 3-1/4" x 4" films
* EMS catalog# 47894-25, holds 25 films per rack
* Quantity: 3
* Condition: used but in nice & clean condition

=Film Racks: Ted Pella Lucite Racks for 3-1/4"x4" films
* Ted Pella catalog# 22706, holds 25 films per rack
* Quantity: 3
* Condition: used but in nice & clean condition

=Film Holders for FEI BioTwin CM120 TEM
* Holds 56 3-1/4"x4" films on designated magazines
* Quantity: 2
* Condition: practically new

=Film Magazines for 3-1/4"x4" sheet films
* Works with FEI BioTwin CM120 TEM Film Holder mentioned above
* Quantity: 73
* Condition: practically new


Thank you very much,

Siva Wu
Research Assistant
Dept. of Advanced Electron Microscopy & Imaging
Div. of Cell Biology & Genetics
House Ear Institute
2100 West 3rd Street
Los Angeles, CA 90057
(213)989-6754
swu-at-hei.org


==============================Original
Headers==============================
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==============================Original Headers==============================
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From: gary-at-gaugler.com
Date: Thu, 25 Mar 2010 00:06:52 -0500
Subject: [Microscopy] help with diagnostic

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

Thanks guys for the many books suggestion for SEM and TEM.


Regards
Fiona Chia


----- Original Message -----
X-from: "Henry Hong" {henry.hong-at-utoronto.ca}
To: {fionacxr-at-tll.org.sg}
Sent: Thursday, March 25, 2010 12:29 AM

I would agree that this is partially true. But I fully
agree that a console with lots of buttons sans PC is
a discrete and historic differentiation from the current
generation of PC-based tools.

I think that much has to do with how the PC interfaces to
the SEM/TEM electronics. Ideally, the PC is simply an
interface of commands...it does not directly control the
tool in real time. To make this clear, if the PC is shut off, does
the tool shut off? It should not. The PC is the GUI interface
to tell the tool what to do. Once told, the tool does not
need the PC. The user could use the PC to monitor and view
what the tool is doing (and visual data is transferred to the
PC GUI). The PC is nothing more than a portal to the tool.
That is how it should be...IMO.

The addition of macros and other automated features that the
PC adds are beneficial but can be fatal if the PC crashes...
which it inevitably will. The PC has two major failure
modes: hard drive failure and mother board failure. Hard
drive failure can be avoided by using RAID 1 but mother board
failure is unlikely to be fully resolved (one maker's RAID is not
necessarily compatible with another's RAID structure--Get Ghost
and routinely do an external backup).

I've experienced PC motherboard failure for Zeiss SEM and
it is a nightmare scenario. But this is not isolated to
Zeiss. The point is well taken that as long as the tool
is constrained for control by the embedded processors (older SEMs), anything
else makes little impact. These systems typically need active
or passive image capture devices--but in the overall scheme of
things, it is a small price to pay. One does not see frequent
updates or patches to the PC GUI on PC-less tools (it does not happen).

The ability to fool around with PC GUIs is rather a job security issue in
contrast to making a solid tool, IMO. Programming is the process of
introducing bugs in software. The more bugs, the more de-bugging
is needed. More justification for programmer/bug persons.

Who are the bug identifiers? Of course, the users. Wonderful...sigh.

gary g.


At 04:15 AM 3/24/2010, you wrote:



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



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From: yu_zongsen-at-yahoo.com
Date: Thu, 25 Mar 2010 04:56:34 -0500
Subject: [Microscopy] EDS quantification in DM

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

Dear All


Our microscope (JEOL 2100F with Digiscan ) is equipped with Inca system for EDS measurement (spot, linescan etc ..). The new version DM (1.71.38) also includes the capabilities for EDS acquisition and analysis, which is really convenient for us to simultaneously obtain EELS and EDS spectra. However, we found that EDS quantification from DM and from Inca system give different results. Does anyone have an idea about this discrepancy ?

Thanks,


Zaoli




==============================Original Headers==============================
8, 25 -- From yu_zongsen-at-yahoo.com Thu Mar 25 04:56:34 2010
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8, 25 -- Date: Thu, 25 Mar 2010 02:56:33 -0700 (PDT)
8, 25 -- From: zongsen yu {yu_zongsen-at-yahoo.com}
8, 25 -- Subject: EDS quantification in DM
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From: protrain-at-emcourses.com
Date: Thu, 25 Mar 2010 06:59:25 -0500
Subject: [Microscopy] EDS quantification in DM

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

Hi

Just one question, have you run a standard sample in each mode where the
content is proven by other means? In this way you would see the way the two
procedures process the same data and the possible errors generated.


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: yu_zongsen-at-yahoo.com [mailto:yu_zongsen-at-yahoo.com]
Sent: 25 March 2010 09:58
To: protrain-at-emcourses.com

Dear All


Our microscope (JEOL 2100F with Digiscan ) is equipped with Inca system for
EDS measurement (spot, linescan etc ..). The new version DM (1.71.38) also
includes the capabilities for EDS acquisition and analysis, which is really
convenient for us to simultaneously obtain EELS and EDS spectra. However, we
found that EDS quantification from DM and from Inca system give different
results. Does anyone have an idea about this discrepancy ?

Thanks,


Zaoli




==============================Original Headers==============================
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==============================Original Headers==============================
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From: protrain-at-emcourses.com
Date: Thu, 25 Mar 2010 08:16:39 -0500
Subject: [Microscopy] EDS quantification in DM

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Hi

Just one question, have you run a standard sample in each mode where the
content is proven by other means? In this way you would see the way the two
procedures process the same data and the possible errors generated.


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide Tel +44
(0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com

-----Original Message-----
X-from: yu_zongsen-at-yahoo.com [mailto:yu_zongsen-at-yahoo.com]
Sent: 25 March 2010 09:58
To: protrain-at-emcourses.com

Dear All


Our microscope (JEOL 2100F with Digiscan ) is equipped with Inca system for
EDS measurement (spot, linescan etc ..). The new version DM (1.71.38) also
includes the capabilities for EDS acquisition and analysis, which is really
convenient for us to simultaneously obtain EELS and EDS spectra. However, we
found that EDS quantification from DM and from Inca system give different
results. Does anyone have an idea about this discrepancy ?

Thanks,


Zaoli




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From: rmcobb-at-crimson.ua.edu
Date: Thu, 25 Mar 2010 20:57:49 -0500
Subject: [Microscopy] viaWWW: Glutaraldehyde sample prep for SEM imaging

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Email: rmcobb-at-crimson.ua.edu
Name: Robin Cobb

Organization: University of Alabama

Title-Subject: [Filtered] Glutaraldehyde sample prep for SEM imaging

Message: I am about to participate in a research cruise where we are
going to be bringing up some coral samples where we want to save the
soft tissue as well as the hard skeleton for SEM imaging. We were
told glutaraldehyde was an option, although I have not worked with it
before. Has anyone worked with this before or have any other
suggestions? We would have to preserve the soft tissue on the boat
before we could bring it back to the lab.

Thank you for your time,
Robin Cobb

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From: rdb-at-cen.dtu.dk
Date: Thu, 25 Mar 2010 21:22:26 -0500
Subject: [Microscopy] viaWWW: 20th General Meeting of the International Mineralogical

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Email: rdb-at-cen.dtu.dk
Name: Rafal E. Dunin-Borkowski

Organization: Center for Electron Nanoscopy Technical University of Denmark

Title-Subject: [Filtered] 20th General Meeting of
the International Mineralogical Association

Message: 20th General Meeting of the International Mineralogical Association
21-27 August 2010
Budapest, Hungary

Scientific session MA91: Advanced transmission electron microscopy methods
Convenors: Rafal E. Dunin-Borkowski, Istv·n DÛdony, Michael Czank
Abstract submission deadline: 6 April 2010

http://www.ima2010.hu/?p=sp3ma
-----------------------------------------------

Many thanks as always,

Rafal E. Dunin-Borkowski
Director, Center for Electron Nanoscopy
Technical University of Denmark
DK-2800 Kongens Lyngby
Denmark
Office: Building 307 Room 101
Tel. +45 4525 6465
Mob +45 2284 0975 rdb-at-cen.dtu.dk
Fax +45 4588 0307 http://www.rafaldb.com/

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From: nizets2-at-yahoo.com
Date: Fri, 26 Mar 2010 02:14:10 -0500
Subject: [Microscopy] viaWWW: Glutaraldehyde sample prep for SEM imaging

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Hi Robin!

Perhaps the best solution to preserve the tissue is to keep it alive?
Is it really impossible to bring a water tank and fill it where the local water and keep it at constant temperature during the cruise?
Apart from the fixative (glutaraldehyde + formaldehyde), other parameters are important to consider during the fixation: pH, temperature and osmoticity for example.
Is it better to perform the fixation at see water osmoticity?
If you can do it, I would consider performing several fixations in  different conditions. Then you can choose the one which works best.

Best regards and have a nice trip

Stephane



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Email: rmcobb-at-crimson.ua.edu
Name: Robin Cobb

Organization: University of Alabama

Title-Subject: [Filtered] Glutaraldehyde sample prep for SEM imaging

Message: I am about to participate in a research cruise where we are
going to be bringing up some coral samples where we want to save the
soft tissue as well as the hard skeleton for  SEM imaging. We were
told glutaraldehyde was an option, although I have not worked with it
before. Has anyone worked with this before or have any other
suggestions? We would have to preserve the soft tissue on the boat
before we could bring it back to the lab.

Thank you for your time,
Robin Cobb

  Login Host: 98.71.104.17
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From: marienti-at-tiscali.it
Date: Fri, 26 Mar 2010 06:39:01 -0500
Subject: [Microscopy] viaWWW: diagrams of the ETEC AUTOSCAN

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Email: marienti-at-tiscali.it
Name: Marco Arienti

Organization: Electron Rescue

Title-Subject: [Filtered] ETEC

Message: I am based in Milano Italy.
I am looking for the diagrams of the ETEC AUTOSCAN "vintage" SEM.
I specially need the diagrams of the power supply (but better to have
all of them) since the +24V is missing on the magnification control
box despite it is present on the filter capacitors.

Thanks in advance for the help

Marco Arienti
www.electronrescue.com


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From: reznik-at-ict.uni-karlsruhe.de
Date: Fri, 26 Mar 2010 06:39:27 -0500
Subject: [Microscopy] viaWWW: SCD 040_SEM_Coater Problem_Solution

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Email: reznik-at-ict.uni-karlsruhe.de
Name: Boris

Organization: Karlsruhe University

Title-Subject: [Filtered] SCD 040_SEM_Coater Problem_Solution

Message: Dear Users,
first of all many thanks for all advices!
Finnaly we found the solutution- the connecting HV cable has been
buckled and therefore there was no current.

With best regards
Boris Reznik

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From: lene.cecilie.hermansen-at-veths.no
Date: Fri, 26 Mar 2010 06:39:50 -0500
Subject: [Microscopy] viaWWW: Localization of the lab

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Email: lene.cecilie.hermansen-at-veths.no
Name: Lene Cecilie Hermansen

Title-Subject: [Filtered] Localization of the lab

Message: Hi Listers

My workplace is moving into new buildings, which
are not yet build. I mentioned to some of the
people involved in this planning process that the
EM-lab need to be situated in the îbasementî, and
then the question came up if it is possible to
make the lab on the ground floor, or if the
vibration in the building will be to disturbing.
The labs I have visited have always been
localized in the bottom of the buildings.

Does anybody have any thoughts?

Regards

Lene Hermansen,
The Norwegian School of Veterinary Science,
Oslo, Norway


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From: ahmad_ds-at-yahoo.com
Date: Fri, 26 Mar 2010 06:40:18 -0500
Subject: [Microscopy] viaWWW: RE: EM texts

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Email: ahmad_ds-at-yahoo.com
Name: Ahmad Ashkaibi

Organization: BAU

Title-Subject: [Filtered] RE: EM texts

Message: Hello everybody,

There are a lot of really good texts by Springer Pub.
As for SEM I recommend the following book:


Scanning Electron Microscopy and X-ray Microanalysis
Author: Joseph Goldstein, Dale E. Newbury, David C. Joy, Charles E.
Lyman, Patrick Echlin, Eric Lifshin, Linda Sawyer, J.R. Michael
ISBN: 0306472929 / 9780306472923
Publisher: Springer Format: Hardback Year: 2003

for TEM, I recommend:

Introduction to Conventional Transmission Electron Microscopy
(Cambridge Solid State Science)
# Publisher: Cambridge University Press
# Number Of Pages: 740
# Publication Date: 2003-04-21
# ISBN / ASIN: 0521620066
# Binding: Hardcover
# Manufacturer: Cambridge University Press

http://gigapedia.com/items:links?id=62940

another TEM textbook for materials engineering:

Transmission Electron Microscopy: A Textbook for Materials Science
By David B. Williams, C. Barry Carter


* Publisher: Springer
* Number Of Pages: 850
* Publication Date: 2009-06-26
* ISBN-10 / ASIN: 038776500X
* ISBN-13 / EAN: 9780387765006

link: http://gigapedia.com/items:links?id=361292

for TEM sample preparation:
Handbook of Sample Preparation for Scanning Electron Microscopy and
X-Ray Microanalysis
By Patrick Echlin


* Publisher: Springer
* Number Of Pages: 200
* ISBN-10 / ASIN: 0387857303
* ISBN-13 / EAN: 9780387857305

link: http://gigapedia.com/items:links?id=274849

Regards,

Ahmad Ashkaibi



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From: oshel1pe-at-cmich.edu
Date: Fri, 26 Mar 2010 07:41:48 -0500
Subject: [Microscopy] Ask-A-Microscopist Establishing TiB2 particle in Optical

Contents Retrieved from Microscopy Listserver Archives
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==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
==========================================================

} } realname - amit kamble
} } Email - amit-at-minexindia.com
} } ORGANIZATION - Minex Metallurgical Co. Ltd.
} } EDUCATION - Graduate College
} } LOCATION - Nagpur, Maharashtra, India
} } SUBJECT_OF_QUESTION - Establishing TiB2 particle in Optical Microscopy

} Reply-To: {amit-at-minexindia.com}
} From: "Amit Kamble" {amit-at-minexindia.com}
} To: "'Philip Oshel'" {oshel1pe-at-cmich.edu}
} Subject: RE: Ask-A-Microscopist
} Date: Fri, 26 Mar 2010 15:29:48 +0530
} Organization: Minex Metallurgical Co. Ltd.
} We are producers of Aluminum Grain Refiner Alloy in
} central India. During establishment of process parameters, we required
} detailed analysis of grain refiner (TiBAl) microstructure. The alloy
} contains Ti-5% B-1% and rest Al, the major phases produced are TiAl3 and
} TiB2. We draw TiBAl grain refiner in wire form. LSM is world leader in
} manufacturing of this alloy.
}
} We require to evaluate the procedure for TiBAl grain refiner - fine
} polishing techniques.
} Presently we are using Buehler Alumina solution, followed by Diamond paste
} of 3micron.
} We are not able to revel the structure properly.
} Kindly help us in elaborating the technique for getting good
} microstructures.
}
} TiAl3 and TiB2 are the two phases under analysis.
} Particle Size :TiAl3 - 30-50 micron
} :TiB2 - 1-2 micron
}
} Some query as:
} How should we polish so that it will revel the lowest particle of 1-2 micron
} ?
} (as I suppose excessive polishing rip of the TiB2 from Al matrix?)
} We need to evaluate the grain size and Ti/B particle distribution in the
} micrograph?
} How could we count the 1-2 micron particle with light microscope?
} The samples are in the form of wire with thickness of 13mm or 9mm.
}
} We also need to evaluate the samples under SEM EDS. Does it need still
} special polishing? We also need to study the TiB2 particles under it.
} Earlier, we tried to study the TiB2 particle under SEM but we faced the
} sample charged difficulty.(i.e. we could not able to any particle structure,
} all screen was bright)
} Please provide some remedy for the same, so that we can analyze the sample
} under SEM easily.
}
} Thanking you
} Regards,
}
} Amit Kamble
} } realname - amit kamble
} } Email - amit-at-minexindia.com
} } ORGANIZATION - Minex Metallurgical Co. Ltd.
} } EDUCATION - Graduate College
} } LOCATION - Nagpur, Maharashtra, India
} } SUBJECT_OF_QUESTION - Establishing TiB2 particle in Optical Microscopy
} } QUESTION - We at minex are producers of Al master alloys in central
} } India. TiBAl Aluminum Alloy contains Ti and B as main additives.
} } Typical grain size is 30-50 micron for TiAl3 and 1-2 micron for TiB2
} } particles.
} } Kindly give us some technical feedback on developing such typical
} micrographs.
}
} --
} Philip Oshel
} Microscopy Facility Supervisor
} Biology Department
} 024C Brooks Hall
} Central Michigan University
} Mt. Pleasant, MI 48859
} (989) 774-3576
} Microscopy Society of America
} Ask a Microscopist
} http://www.microscopy.org/microscopy/ask.cfm
}

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From: lherault-at-bu.edu
Date: Fri, 26 Mar 2010 08:20:42 -0500
Subject: [Microscopy] Ask-A-Microscopist Establishing TiB2 particle in

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

Buehler Co. itself is a good resource. If they contact Beuhler, they will
probably get the help/guidance that they need. I'm a bit surprised more
folks don't turn first to the supplier/manufacturer of their equipment when
they have problems. Most companies want to help their customers/potential
customers because a happy customer is a repeat customer.

So the question is, have they contacted Buehler for help in setting up a
protocol?

After polishing, some kind of etchant may help reveal particles for SEM.

Ron L

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Friday, March 26, 2010 8:44 AM
To: lherault-at-bu.edu

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
==========================================================

} } realname - amit kamble
} } Email - amit-at-minexindia.com
} } ORGANIZATION - Minex Metallurgical Co. Ltd.
} } EDUCATION - Graduate College
} } LOCATION - Nagpur, Maharashtra, India
} } SUBJECT_OF_QUESTION - Establishing TiB2 particle in Optical Microscopy

} Reply-To: {amit-at-minexindia.com}
} From: "Amit Kamble" {amit-at-minexindia.com}
} To: "'Philip Oshel'" {oshel1pe-at-cmich.edu}
} Subject: RE: Ask-A-Microscopist
} Date: Fri, 26 Mar 2010 15:29:48 +0530
} Organization: Minex Metallurgical Co. Ltd.
} We are producers of Aluminum Grain Refiner Alloy in
} central India. During establishment of process parameters, we required
} detailed analysis of grain refiner (TiBAl) microstructure. The alloy
} contains Ti-5% B-1% and rest Al, the major phases produced are TiAl3 and
} TiB2. We draw TiBAl grain refiner in wire form. LSM is world leader in
} manufacturing of this alloy.
}
} We require to evaluate the procedure for TiBAl grain refiner - fine
} polishing techniques.
} Presently we are using Buehler Alumina solution, followed by Diamond paste
} of 3micron.
} We are not able to revel the structure properly.
} Kindly help us in elaborating the technique for getting good
} microstructures.
}
} TiAl3 and TiB2 are the two phases under analysis.
} Particle Size :TiAl3 - 30-50 micron
} :TiB2 - 1-2 micron
}
} Some query as:
} How should we polish so that it will revel the lowest particle of 1-2
micron
} ?
} (as I suppose excessive polishing rip of the TiB2 from Al matrix?)
} We need to evaluate the grain size and Ti/B particle distribution in the
} micrograph?
} How could we count the 1-2 micron particle with light microscope?
} The samples are in the form of wire with thickness of 13mm or 9mm.
}
} We also need to evaluate the samples under SEM EDS. Does it need still
} special polishing? We also need to study the TiB2 particles under it.
} Earlier, we tried to study the TiB2 particle under SEM but we faced the
} sample charged difficulty.(i.e. we could not able to any particle
structure,
} all screen was bright)
} Please provide some remedy for the same, so that we can analyze the sample
} under SEM easily.
}
} Thanking you
} Regards,
}
} Amit Kamble
{ {SNIP} }
}
} --
} Philip Oshel
} Microscopy Facility Supervisor
} Biology Department
} 024C Brooks Hall
} Central Michigan University
} Mt. Pleasant, MI 48859
} (989) 774-3576
} Microscopy Society of America
} Ask a Microscopist
} http://www.microscopy.org/microscopy/ask.cfm
}



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13, 23 -- References: {201003261244.o2QCiAVU004876-at-ns.microscopy.com}
13, 23 -- Subject: RE: [Microscopy] Ask-A-Microscopist Establishing TiB2 particle in Optical
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From: kiss-at-demet.ufrgs.br
Date: Fri, 26 Mar 2010 08:51:22 -0500
Subject: [Microscopy] Re: viaWWW: Localization of the lab

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

Hello Lene:
We operated a Philips XL20 for many years in the 6th floor of a
building by a street of quite a heavy traffic with no problems unless
you wanted to go above 10.000 X, then vibration sometimes made work
impossible above this magnification.
My 0,2 cents.....
Regards
kiss

Francisco José Kiss
Dpto. de Metalurgia
Universidade Federal do Rio Grande do Sul
Brazil


---------- Original Message -----------
X-from: lene.cecilie.hermansen-at-veths.no
To: kiss-at-demet.ufrgs.br


} Email: lene.cecilie.hermansen-at-veths.no
} Name: Lene Cecilie Hermansen
}
} Title-Subject: [Filtered] Localization of the lab
}
} Message: Hi Listers
}
} My workplace is moving into new buildings, which
} are not yet build. I mentioned to some of the
} people involved in this planning process that the
} EM-lab need to be situated in the îbasementî, and
} then the question came up if it is possible to
} make the lab on the ground floor, or if the
} vibration in the building will be to disturbing.
} The labs I have visited have always been
} localized in the bottom of the buildings.
}
} Does anybody have any thoughts?
}
} Regards
}
} Lene Hermansen,
} The Norwegian School of Veterinary Science,
} Oslo, Norway
}
} Login Host: 158.36.103.211
} -

==============================Original Headers==============================
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5, 20 -- To: lene.cecilie.hermansen-at-veths.no, microscopy-at-microscopy.com
5, 20 -- Subject: Re: [Microscopy] viaWWW: Localization of the lab
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From: eschumacher-at-mccrone.com
Date: Fri, 26 Mar 2010 08:54:50 -0500
Subject: [Microscopy] Ask-A-Microscopist Establishing TiB2 particle in

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

I agree that equipment suppliers can be valuable resources. When a client needs to outsource work, an equipment manufacturer may have an applications lab that can assist with sample prep and analysis. Vendors may also be able to share information about labs where they've placed their equipment. If these are consulting labs or centralized characterization labs in academic institutions, the client may be able to outsource the work to them. When faced with a request for a technique that we don't do here, I've often recommended contacting equipment suppliers, along with referrals to other analytical laboratories.

Elaine

*********************************************************************
Elaine F.Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com



*********************************************************************
This message and any attachments are solely for the
intended recipient. If you are not the intended recipient,
disclosure, copying, use or distribution of the information
included in this message is prohibited.
*********************************************************************

-----Original Message-----
X-from: lherault-at-bu.edu [mailto:lherault-at-bu.edu]
Sent: Friday, March 26, 2010 8:29 AM
To: Elaine F. Schumacher

Buehler Co. itself is a good resource. If they contact Beuhler, they will
probably get the help/guidance that they need. I'm a bit surprised more
folks don't turn first to the supplier/manufacturer of their equipment when
they have problems. Most companies want to help their customers/potential
customers because a happy customer is a repeat customer.

So the question is, have they contacted Buehler for help in setting up a
protocol?

After polishing, some kind of etchant may help reveal particles for SEM.

Ron L

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Friday, March 26, 2010 8:44 AM
To: lherault-at-bu.edu

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
==========================================================

} } realname - amit kamble
} } Email - amit-at-minexindia.com
} } ORGANIZATION - Minex Metallurgical Co. Ltd.
} } EDUCATION - Graduate College
} } LOCATION - Nagpur, Maharashtra, India
} } SUBJECT_OF_QUESTION - Establishing TiB2 particle in Optical Microscopy

} Reply-To: {amit-at-minexindia.com}
} From: "Amit Kamble" {amit-at-minexindia.com}
} To: "'Philip Oshel'" {oshel1pe-at-cmich.edu}
} Subject: RE: Ask-A-Microscopist
} Date: Fri, 26 Mar 2010 15:29:48 +0530
} Organization: Minex Metallurgical Co. Ltd.
} We are producers of Aluminum Grain Refiner Alloy in
} central India. During establishment of process parameters, we required
} detailed analysis of grain refiner (TiBAl) microstructure. The alloy
} contains Ti-5% B-1% and rest Al, the major phases produced are TiAl3 and
} TiB2. We draw TiBAl grain refiner in wire form. LSM is world leader in
} manufacturing of this alloy.
}
} We require to evaluate the procedure for TiBAl grain refiner - fine
} polishing techniques.
} Presently we are using Buehler Alumina solution, followed by Diamond paste
} of 3micron.
} We are not able to revel the structure properly.
} Kindly help us in elaborating the technique for getting good
} microstructures.
}
} TiAl3 and TiB2 are the two phases under analysis.
} Particle Size :TiAl3 - 30-50 micron
} :TiB2 - 1-2 micron
}
} Some query as:
} How should we polish so that it will revel the lowest particle of 1-2
micron
} ?
} (as I suppose excessive polishing rip of the TiB2 from Al matrix?)
} We need to evaluate the grain size and Ti/B particle distribution in the
} micrograph?
} How could we count the 1-2 micron particle with light microscope?
} The samples are in the form of wire with thickness of 13mm or 9mm.
}
} We also need to evaluate the samples under SEM EDS. Does it need still
} special polishing? We also need to study the TiB2 particles under it.
} Earlier, we tried to study the TiB2 particle under SEM but we faced the
} sample charged difficulty.(i.e. we could not able to any particle
structure,
} all screen was bright)
} Please provide some remedy for the same, so that we can analyze the sample
} under SEM easily.
}
} Thanking you
} Regards,
}
} Amit Kamble
{ {SNIP} }
}
} --
} Philip Oshel
} Microscopy Facility Supervisor
} Biology Department
} 024C Brooks Hall
} Central Michigan University
} Mt. Pleasant, MI 48859
} (989) 774-3576
} Microscopy Society of America
} Ask a Microscopist
} http://www.microscopy.org/microscopy/ask.cfm
}



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From: richard.ross-at-allisontransmission.com
Date: Fri, 26 Mar 2010 09:02:09 -0500
Subject: [Microscopy] Re: viaWWW: Localization of the lab

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

Lene,

Realize that all structures have natural vibration frequencies. The
vibration level in a basement, where the floor is most likely poured
concrete on undisturbed soil, is probably less in magnitude and of a lower
frequency than what would be in the building floors suspended above by the
building structure. The vibration of equipment within the building only
adds to these vibrations and issues.

The basic goal, as I understand things, is to place the microscope on a
sufficiently massive base that the fundamental vibration frequency of the
base is well below the fundamental frequency of the microscope structure.
Thus the base tends to dampen any vibration frequencies of potential harm
to the scope.

In a basement, the ideal would be to excavate a large amount of soil and
pour a large block (mass) of concrete for the microscope to ultimately sit
upon. This block should be mechanically isolated from the surrounding
concrete floor and building structure (not solidly connected).

On upper floors, this large mass is difficult to achieve for some obvious
reasons of weight and size. There one resorts to vibration isolation
platforms, either passive (moderately costly) or active closed-loop
systems (quite costly).

My personal experience is with a microscope on the main floor of a 1940's
building (thin concrete slab on soil) and I have vibration issues coming
from the building and plant equipment. We have installed a passive
air-sprung isolation platform ($7K US) that helps to some extent but isn't
a cure-all. If I were given the choice between a basement and upper floor,
the basement would definitely be the desirable location. If you are in the
early stage of construction and can get a separate isolation block poured,
all the better.

Rick





lene.cecilie.hermansen-at-veths.no
03/26/2010 07:56 AM
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Email: lene.cecilie.hermansen-at-veths.no
Name: Lene Cecilie Hermansen

Title-Subject: [Filtered] Localization of the lab

Message: Hi Listers

My workplace is moving into new buildings, which
are not yet build. I mentioned to some of the
people involved in this planning process that the
EM-lab need to be situated in the îbasementî, and
then the question came up if it is possible to
make the lab on the ground floor, or if the
vibration in the building will be to disturbing.
The labs I have visited have always been
localized in the bottom of the buildings.

Does anybody have any thoughts?

Regards

Lene Hermansen,
The Norwegian School of Veterinary Science,
Oslo, Norway


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From: protrain-at-emcourses.com
Date: Fri, 26 Mar 2010 09:27:50 -0500
Subject: [Microscopy] viaWWW: Localization of the lab

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

Lene,

I have been mostly in basements for TEM and SEM but I have also been on second and third floors and had no problems. At the moment I am in a one floor building which has a thick concrete floor AND there are windows in the hallways outside my lab rooms. What joy to see that the world still exists even if it is raining! I have used an old MT2 ultra-microtome on a bench here and only have a slight vibration problem if a really big truck passes by outside (infrequently).

In the basement at my previous position (in a big city) there were frequent vibrations from large trucks hitting the parking lot curb outside my building (making deliveries) and of course the jack hammers when the street or sidewalk needed to be torn up for checking leaking pipes or surface replacement. Most of this type work is done early in the day so I'd wait until 2:30 or so to start imaging or thin sectioning.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E - Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-480-6560
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov} {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not represent the NIH. This message is not confidential and can be freely shared and reproduced.
________________________________
X-from: {lene.cecilie.hermansen-at-veths.no}
Reply-To: {lene.cecilie.hermansen-at-veths.no}

Hi Lene

Others have talked about vibration, which was the point of your question,
but there are these days more difficult problems!

If you intend to become involved with the high performance field emission
SEM that are now available from all of the manufacturers, you should play
particular attention to magnetic fields. Whilst there are a number of
pretty good anti vibration systems available for minor problems, there in my
experience does not seem to be devices for compensating magnetic field that
are anything like as good!

So keep an eye on high current carrying cables, lifts, local heavy current
drawing machinery etc, as well as giving due consideration to vibration.

I once put a TEM on the 6th floor of a building, taking great care to be
involved with the design of the building to optimise the microscope
position. I noted that the 7th floor was the animal house, however, not
until we installed the microscope did I find that at each end of the animal
house were fans 7 feet across (air vibration, floor vibration and magnetic
fields). Fortunately for me I understand they have to this day not used the
fans.

Good luck

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com




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Email: lene.cecilie.hermansen-at-veths.no
Name: Lene Cecilie Hermansen

Title-Subject: [Filtered] Localization of the lab

Message: Hi Listers

My workplace is moving into new buildings, which
are not yet build. I mentioned to some of the
people involved in this planning process that the
EM-lab need to be situated in the îbasementî, and
then the question came up if it is possible to
make the lab on the ground floor, or if the
vibration in the building will be to disturbing.
The labs I have visited have always been
localized in the bottom of the buildings.

Does anybody have any thoughts?

Regards

Lene Hermansen,
The Norwegian School of Veterinary Science,
Oslo, Norway


Login Host: 158.36.103.211
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From: contact-at-integrityscientific.com
Date: Fri, 26 Mar 2010 10:14:54 -0500
Subject: [Microscopy] viaWWW: Localization of the lab

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

Hi Lene,
contact the microscope manufacturer and ask for their intallation requirements. This should list the vibration requirements, as well as other important things like temperature stability and e-m field limits. Give this to the architects, so that it forms part of their specifications for the building.

If you have high-spec microscopes, watching their reaction can be quite interesting!

Good luck

Richard

PS something you may need to specify to meet e-m field specs, which is easy to miss - lightning conductors must not form ground loops. This means if there are multiple conductors they must be connected at the bottom, not the top (opposite to the usual way of doing things).

This Question/Comment was submitted to the Microscopy Listserver
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Email: lene.cecilie.hermansen-at-veths.no
Name: Lene Cecilie Hermansen

Title-Subject: [Filtered] Localization of the lab

Message: Hi Listers

My workplace is moving into new buildings, which
are not yet build. I mentioned to some of the
people involved in this planning process that the
EM-lab need to be situated in the îbasementî, and
then the question came up if it is possible to
make the lab on the ground floor, or if the
vibration in the building will be to disturbing.
The labs I have visited have always been
localized in the bottom of the buildings.

Does anybody have any thoughts?

Regards

Lene Hermansen,
The Norwegian School of Veterinary Science,
Oslo, Norway


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From: donovan-at-uoregon.edu
Date: Fri, 26 Mar 2010 12:46:29 -0500
Subject: [Microscopy] Re: viaWWW: Localization of the lab

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

All,
It should be noted that manufacturers have significant pressure to
develop "lowest common denominator" environmental performance
specifications because they don't want to lock themselves out of the
competition with regard to a prospective customer's lab space that
happens to be less than ideal.

When we designed our underground lab space for nano-characterization
in Oregon the first thing the architects asked for were the vendor's
"cut sheets" for each instrument. But I explained to them the above
mentioned concern and convinced them that these vendor installation
specs were only a starting point for vibration, acoustics, EMI,
temperature control, etc. What we tried to do was design the entire
analytical section of the building for ultimate performance based on
significantly exceeding the specs of the most sensitive instrument we
planned on (the FEI Titan TEM).

That meant designing the floor poured directly on bed rock 20 feet
below the surface, oversize round ventilation ducting for low speed
non-turbulent air flow, specially engineered high sensitivity
temperature control systems (based on off the shelf components),
electrically isolated rebar in the slabs, all steel electrical
conduit and extra shielding for all electrical panels and devices, 12
volt DC lighting for instrument operation, dedicated low noise earth
grounds, all building transformers, water handling, compressors,
exhaust and supply fans all located in other nearby buildings and
plumbed over reasonable distances to our isolated building which has
a turf roof and in which we are in the basement sitting on Eugene
Formation bed rock.

In fairness I should point out that this Eugene Formation bedrock is
serendipitously a perfect foundation material for such a purpose due
to it's porousity and high damping coefficient. In fact, one
vibration consultant claimed that this site, in the middle of our
campus, was the second quietest site he had ever measured in his
career and when we asked what was the quietest site he said: "well,
there's this mountain top in New Mexico that is 160 miles from the
nearest city... that's a little bit quieter".

What we ended up with was beautiful, but in space of my career it was
a one of a kind perfect storm of support from the administration
(Rich Linton is vice-provost of research here), a dedicated group of
faculty that wanted to do something significant for a shared user
facility, generous private funding (Lorry I. Lokey) and economic
development support from the state (ONAMI), an excellent contractor
(Lease Crutcher Lewis in Portland) and architect (SRG in Portland),
and very talented mechanical, electrical and HVAC engineers
(Balzhiser & Hubbard Engineers in Eugene) that understood that "value
engineering" has a real downside.

More technical stuff can be found here, but let me just point with
some pride that our floors exceed NIST-A vibration levels by several factors:

http://camcor.uoregon.edu/fac_tour.shtml

The result is that all the instruments in our facility exceed their
factory performance specifications in multiple ways (or if they
don't, it sure isn't due to the building environment!). For myself as
an EPMA guy, it is the 0.3 degrees F temperature control that gives
my Bragg spectrometers the reproducibility I've always dreamed of.

How much did this 30K sq feet building cost? About half the cost of
all the instruments housed in it, and well worth it I might add.

I apologize in advanced for "tooting our own horn" a bit here, but
some of you might be in Portland this summer for M&M and I thought
afterwards some of you might want to take a short drive down the
freeway (about 2 hours) to see our Eugene CAMCOR facility. I won't be
at the CAMCOR facility the Friday after M&M concludes (I'll be there
all day Saturday August 7th for our annual EPMA workshop), but most
of our technical/scientific staff will be there and I'm sure they
would be pleased to show you around.

You can contact Paula Matano, Kurt Langworthy, Steve Golledge or
Sujing Xie if you are interested in visiting Eugene this summer (or
any time you are in the area for that matter). Our contact info is here:

http://camcor.uoregon.edu/contact.shtml

Hope to see you all in Portland this summer!
John Donovan

Almost forgot- the EPMA workshop link is here:

http://camcor.uoregon.edu/includes/events/event_7aug2010.html


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From: bioanalytics-at-ibilabs.com
Date: Fri, 26 Mar 2010 18:34:52 -0500
Subject: [Microscopy] viaWWW: Uranyl Glass

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

Hi Everybody:
I made this kind of metallography some 30+ years ago so it may be a
little out of date.
Our material was made of Al 99,99% Ti was aded as F6TiK2, B was added
as F4BH individually or together. Another form to include this
elements was under the form of a comertial product "FOSECO NUCLEANT II".
These mateials were added to Al at 800 °C. The analised material was
as cast.
Three techniques were used:
1) The free surface of the ingot was lightly polished electrolitically
in a cell to clean it, and atacked with a Disa Electropol aparatus.
2) The free surface of the ingot suffered only an anodic oxidation.
3)Cuts of the ingots were mechanically polished, then electrolitically
in a cell and structure revealed by anodic oxidation.

Observation on the microscope gives the best results with phase
contrast, Nomarsky system. Also dark field ilumination gives good
results. Of course clear field is OK.

Mechanical polish: after grinding to emery paper 600,finally polished
with 7 microns diamond paste. This last step gives later a better
result but can be left out.

Electrolitic polishing in cell was made with Al cathode temperature
kept under 10°C, time from 5 to 10 minutes cell voltage ,open
circuit, around 28 volts(depends of the power supply characteristics),
composition of the electrolite was:
Percloric acid 10%
Glicerine 3%
Butilcellosolve 87% all by volume
(Butilcellosolve is also known as etileneglicol-monobutil eter or
butilic glicol).

DISA Electropol A2 electrolite
Percloric acid 78 ml. (added last)
Distiled water 120 ml.
Ethanol 700 ml.
Butilcellosolve 100 ml.
When used to polish current is about 1A/cm2, for attack 60 mA/cm2
temperature should be kept under 25°C

Anodic oxidation was made in a cell, cathode was stainless steel
(18/8), Voltage about 10 to 18 V. time about 1 minute.
Electrolite is 10% sulphuric acid in water.

This oxide can be scratched with a sharp point then put into a 5%
solution of mercury cloride in water. The oxide layer with the Al3Ti
particles attached will be separated and put into a TEM for difraction
or EDS. Our paricles were too big for difraction and had no EDS.

The "normal" specimens were analized in a CAMECA WDS with good results
as for Al3TI but not for Boron ;-).
Hope this is of help.
Sorry for the bandwith.
Best of luck
Regards
kiss

Francisco José Kiss
Dpt. of Metallurgy
Universidade Federal de Rio Grande do Sul
Porto Alegre
Brazil
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Email: bioanalytics-at-ibilabs.com
Name: Alex Besenyo PhD

Organization: IBI Labs

Title-Subject: [Filtered] Uranyl Glass

Message: Good Day,
Recently I recieved a request for information regarding Uranyl Glass
related to fluorescence intensity.

Quote: "The average NADH fluoresence intensity of each surface area
is related to the fluoresence intensity of a fluoresence calibration
glass(uranyl) and is expressed in arbitrary units."

Can anyone shed any light on what is uranyl glass?

Thank you
Alex



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From: PhillipsT-at-missouri.edu
Date: Fri, 26 Mar 2010 19:52:48 -0500
Subject: [Microscopy] viaWWW: Uranyl Glass

Contents Retrieved from Microscopy Listserver Archives
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It is a glass that has a uranyl salt incorporated into it. The slides are generally yellowish if I remember correctly. I bought some years ago but not sure where. They give off a strong fluorescent signal that if resistant to fading so they can be used as a calibration aid. The trick is measuring the "real" signal of the unknown without it fading.

Thomas E. Phillips, Ph.D.
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
Biological Sciences
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (voice)
573-882-0123 (fax)

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Email: bioanalytics-at-ibilabs.com
Name: Alex Besenyo PhD

Organization: IBI Labs

Title-Subject: [Filtered] Uranyl Glass

Message: Good Day,
Recently I recieved a request for information regarding Uranyl Glass
related to fluorescence intensity.

Quote: "The average NADH fluoresence intensity of each surface area
is related to the fluoresence intensity of a fluoresence calibration
glass(uranyl) and is expressed in arbitrary units."

Can anyone shed any light on what is uranyl glass?

Thank you
Alex



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From: Frank_Karl-at-lincolnelectric.com
Date: Sat, 27 Mar 2010 07:17:05 -0500
Subject: [Microscopy] Re: viaWWW: Uranyl Glass

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

In the days before reflected fluorescence, when you used high power Hg
burners to produce UV which was focused through the quartz slide and into
the objectives, cubes of uranyl glass were used. They demonstrated that
you corrected alined the sub-stage optics and were getting UV, the shape of
the illumination cone and its location as well as the location and size of
the cross-over point.

I remenber yellow or orange plastic shields used to protect your eyes from
UV that might creep around the stage and microscope body. I used one at
Goodyear Tire, but I and anyone else was never happy about it and for one,
I'm glad to see them gone.

stay safe.....
Frank



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abs.com
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03/26/2010 07:43 frank_karl-at-lincolnelectric.com
PM cc

Subject
Please respond to [Microscopy] viaWWW: Uranyl Glass
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Email: bioanalytics-at-ibilabs.com
Name: Alex Besenyo PhD

Organization: IBI Labs

Title-Subject: [Filtered] Uranyl Glass

Message: Good Day,
Recently I recieved a request for information regarding Uranyl Glass
related to fluorescence intensity.

Quote: "The average NADH fluoresence intensity of each surface area
is related to the fluoresence intensity of a fluoresence calibration
glass(uranyl) and is expressed in arbitrary units."

Can anyone shed any light on what is uranyl glass?

Thank you
Alex



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From: zaluzec-at-aaem.amc.anl.gov
Date: Sat, 27 Mar 2010 08:59:44 -0500
Subject: [Microscopy] Adminisitrivia: Text Books on File Sharing Sites

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

Colleagues

It has come to my attention that during the recent thread of posting
about text books a link was supplied to a site called http://4shared.com

This site allows users to upload documents for file sharing, much like
some of the music sites did a few years ago. Unfortunately, just
as the music industry saw instances of abuse, the same holds
here and the are illegal postings of copyrighted books therein.

Just as we should protect music copyright holders we should also
protect as a community our book authors. I have contacted the site
operators and
have informed them that copyrighted EM books are listed on their
WWW site and that these documents should be removed. The
book appears to have been scanned and uploaded by someone
not associated with the Microscopy Listserver.

Please remember, the authors of all books, have put in a
great amount of effort to write these text books and rightly
deserve to have the books purchased through legal channels.

I have edited the archives and removed the links to the file
sharing site, however, the remainder of the posting
is fine and provides links to reviews of the EM text books
http://gigipedia.com.

Your Friendly Neighborhood SysOp
Nestor

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From: lene.cecilie.hermansen-at-nvh.no
Date: Mon, 29 Mar 2010 08:12:01 -0500
Subject: [Microscopy] viaWWW: EM-lab location

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Email: lene.cecilie.hermansen-at-nvh.no
Name: Lene Cecilie Hermansen

Organization: The Norwegian School of Veterinary Science

Title-Subject: [Filtered] EM-lab location

Message: Thank you all for the great response I got for the question
on where to locate the EM-lab. I learned that there are so many
variables; the architects have a big job finding the best solution. I
am going to make a resume of a lot of your answers, talk to the
manufacturer and other instances, and hopefully the resulting EM-lab
will work very well.

Lene Hermansen
The Norwegian School of Veterinary Science



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9, 11 -- Subject: viaWWW: EM-lab location
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From: lene.cecilie.hermansen-at-nvh.no
Date: Mon, 29 Mar 2010 08:33:21 -0500
Subject: [Microscopy] viaWWW: EM-lab location

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Email: lene.cecilie.hermansen-at-nvh.no
Name: Lene Cecilie Hermansen

Organization: The Norwegian School of Veterinary Science

Title-Subject: [Filtered] EM-lab location

Message: Thank you all for the great response I got for the question
on where to locate the EM-lab. I learned that there are so many
variables; the architects have a big job finding the best solution. I
am going to make a resume of a lot of your answers, talk to the
manufacturer and other instances, and hopefully the resulting EM-lab
will work very well.

Lene Hermansen
The Norwegian School of Veterinary Science



Login Host: 158.36.103.212
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==============================Original Headers==============================
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From: Frank_Karl-at-lincolnelectric.com
Date: Mon, 29 Mar 2010 10:17:18 -0500
Subject: [Microscopy] 3rd party service for JEOL SEMs

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


Good Morning Everyone,
I've been asked to find a third party service provider for routine and
emergency service for our JEOL 5800 SEM with the JEOL backscatter detector
and Video Measurement Scaler. We are interested in two yearly PM visits
and emergency service.

Please feel free to contact me directly or share your experience with the
community.

Frank Karl
Microscopist
Lincoln Electric
Cleveland OH
216-383-2090


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From: nyilmaz-at-mersin.edu.tr
Date: Mon, 29 Mar 2010 14:59:36 -0500
Subject: [Microscopy] Help for TEM maintenance

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

Dear Colleagues,

We're using a JEOL JEM 1011 TEM in our Histology Dept. since Jan. 2005. The
microscope and its ancillary devices (rotary pump, chiller, UPS etc) were on
for 24 hours almost all this time. My question is: how frequently we should
change the rotary pump oil and also which maintenances do you recommend we
should performed periodically?
Best regards.

Dr. Necat Yilmaz
Mersin University
School of Medicine
Histology & Embryology Dept.


==============================Original Headers==============================
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From: doc.vrdoljak-at-gmail.com
Date: Mon, 29 Mar 2010 16:00:27 -0500
Subject: [Microscopy] image analysis software

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

Hello,
I regularly use Image pro Plus (version 6), for image analysis. I was
wondering if anyone can recommend something else, preferably more
reliable and with better documentation and support for its use.

Thanks, any comments greatly appreciated.

==============================Original Headers==============================
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2, 30 -- Message-ID: {fb272961003291400l6114e884n163c2eeac6dd3428-at-mail.gmail.com}
2, 30 -- Subject: image analysis software
2, 30 -- From: Gordon Vrdoljak {doc.vrdoljak-at-gmail.com}
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From: jsiegmund-at-7thwavelabs.com
Date: Mon, 29 Mar 2010 16:28:12 -0500
Subject: [Microscopy] image analysis software

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

We are just looking into getting a new image analysis program.
So far, we really like Visiomorph by Visiopharm.

Joachim,

Seventhwavelabs, St. Louis

-----Original Message-----
X-from: doc.vrdoljak-at-gmail.com [mailto:doc.vrdoljak-at-gmail.com]
Sent: Monday, March 29, 2010 4:08 PM
To: Joachim Siegmund

Hello,
I regularly use Image pro Plus (version 6), for image analysis. I was
wondering if anyone can recommend something else, preferably more
reliable and with better documentation and support for its use.

Thanks, any comments greatly appreciated.

==============================Original
Headers==============================
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This communication is intended solely for the use of the addressee and may contain information that is legally privileged, confidential or exempt from disclosure. If you are not the intended recipient, please note that any dissemination, distribution, or copying of this communication is strictly prohibited. Anyone who receives this message in error should notify the sender immediately and delete it from his or her computer


==============================Original Headers==============================
12, 30 -- From jsiegmund-at-7thwavelabs.com Mon Mar 29 16:28:11 2010
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From: beth-at-plantbio.uga.edu
Date: Mon, 29 Mar 2010 17:07:13 -0500
Subject: [Microscopy] Reichert-Jung 2800 cryostat

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

Hi all,
Does anyone have a Reichert-Jung 2800 cryostat gathering dust? We need
a drive board for a 2800 so if you have one you can spare or would
sell please let me know. Thanks!
Beth

} Beth,
} The answer is what I expected it to be. The drive board for the 2800
} is no longer available new or refurbished. The next option is to try
} to find one laying around somewhere and worse case scenario it is a
} manual cryostat. I would not expect much luck with somebody having
} one collecting dust on a shelf. Sorry I don't have better news.
}
} David
} David A. Benjamin
} DBMS
} DB MicroService, Inc.
} (P) 770.904.4848 (P) 1.888.904.4848
} (F) 770.904.4855 (F) 1.888.902.4855
} www.dbmicroservice.com
} www.dbsms.biz


**********************************************************************
Beth Richardson
Electron Microscopy Lab Coordinator
Plant Biology Department
Miller Plant Sciences Bldg
120 Carlton Street
University of Georgia
Athens, GA 30602-7271

http://www.plantbio.uga.edu/emlab/

Phone - (706) 542-1790 & FAX - (706) 542-1805

"Between the two evils,
I always pick the one I never tried before". Mae West (1893-1980)
*******************************************************************

"And it's only the giving that makes you what you are".
Wond'ring Aloud, Jethro Tull (Aqualung)

***************************************************************************
The Friends of the Marine Institute - Join Today!
www.friendsofugami.com






==============================Original Headers==============================
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From: khrach-at-mediacy.com
Date: Mon, 29 Mar 2010 17:18:29 -0500
Subject: [Microscopy] image analysis software

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

Dear Gordon,

I'm sorry to hear that you're considering leaving Image-Pro. Over the years, our customers have requested better training & support tools. In the past couple of years, we have been working to develop improved educational tools for our software users. New tools include:

Imaging Webinars -
Each month we offer free online webinars on topics including Introduction to Image Analysis, Image Deconvolution, Macro Recording, and 3D Rendering. Next week we have a webinar on Counting & Sorting Objects.
http://www.mediacy.com/index.aspx?page=WebinarCalendar

Image-Pro Video Tutorials -
We've created a series of quick 3-5 minute tutorial videos on frequently used Image-Pro tools including stitching, thresholding, image alignment, background correction and more. More are added each month.
http://www.mediacy.com/index.aspx?page=IPPTours

Solutions Zone -
We've recently updated our Solutions Zone macro site, where users can download, upload, and share macros. We've made it easier to search and sort for macros and will be adding additional macros in the coming weeks.
http://www.mediacy.com/index.aspx?page=SolutionsZone

I hope you'll take some time to visit some of these new resources and send us your feedback on how we can continue to improve our support tools. Your opinion is important to us.

Regards,
Kathy Hrach

Kathy Hrach
Product Manager
Media Cybernetics
4340 East-West Hwy, Suite 400
Bethesda, MD 20814
Phone: 301-495-3305 ext.260
Email: khrach-at-mediacy.com
www.mediacy.com

NEW Image Pro 7.0.1 is out! http://support.mediacy.com/ipp701.asp


-----Original Message-----
X-from: jsiegmund-at-7thwavelabs.com [mailto:jsiegmund-at-7thwavelabs.com]
Sent: Monday, March 29, 2010 5:34 PM
To: Hrach, Kathy

We are just looking into getting a new image analysis program.
So far, we really like Visiomorph by Visiopharm.

Joachim,

Seventhwavelabs, St. Louis

-----Original Message-----
X-from: doc.vrdoljak-at-gmail.com [mailto:doc.vrdoljak-at-gmail.com]
Sent: Monday, March 29, 2010 4:08 PM
To: Joachim Siegmund

Hello,
I regularly use Image pro Plus (version 6), for image analysis. I was
wondering if anyone can recommend something else, preferably more
reliable and with better documentation and support for its use.

Thanks, any comments greatly appreciated.

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From: Gregory.Hendricks-at-ns.microscopy.com
Date: Mon, 29 Mar 2010 17:37:06 -0500
Subject: [Microscopy] viaWWW: Position for Electron Microscopy Technician

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Email: Gregory.Hendricks
Name: Greg Hendricks

Organization: UMass Medical School

Title-Subject: [Filtered] Job Posting

Message: University of Massachusetts Medical School, Core Electron
Microscopy Facility
Position for Electron Microscopy Technician

The Department of Cell Biology, Core EM Facility, seeks a Research
Assistant to assist in ultrastructural research. The Research
Assistant will be joining a dynamic laboratory and will be
responsible for projects involving transmission electron microscopy
and scanning electron microscopy including the preparation of tissue,
ultramicrotomy and ultrastructural immunocytochemistry, and
collection and analysis of data. The position may be part time (50%
or higher) to full time (100%).

Qualifications:

Requires a Bachelor of Science or higher degree in biology or related
scientific area. Must have demonstrable skills and experience in all
routine aspects of electron microscopy. Must be highly organized
with excellent communication and computer skills, and have an ability
to work independently as well as in a team. Additional skills in
histology, graphics, digital imaging and statistics would be
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From: protrain-at-emcourses.com
Date: Tue, 30 Mar 2010 04:34:44 -0500
Subject: [Microscopy] Help for TEM maintenance

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

Hi Necat

To enhance their efficiency and their life we would suggest you change the
rotary pump fluid at least once a year. The fluid is performing a number of
tasks (1) Lubrication (2) Anti corrosion (3) assisting with the vacuum
seal. If you have a compressed air driven system with its own compressor
its fluid should also be changed, but most of all the tank should be drained
of the moisture which will have built up inside (compressing air produces
moisture).

Other areas requiring attention are the "O" ring on the specimen exchange
rod, the specimen seat(s) and possibly the objective and condenser movable
apertures. If astigmatism levels are high or you are able to see dirt on
the apertures when they are visualised during operation they need cleaning
(high vacuum heating) or replacing.

If you are regularly using 100kV every few years the gun chamber needs a
good clean as the build up of contamination will cause more micro discharge.

Over a period of up to five years the viewing screen(s) will degrade through
contamination (depends on the instrument manufacturer) and should be
replaced.

In all of the above check out the instruction manual prior to making any
attempt to carry out any action.

Good luck


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: nyilmaz-at-mersin.edu.tr [mailto:nyilmaz-at-mersin.edu.tr]
Sent: 29 March 2010 21:01
To: protrain-at-emcourses.com

Dear Colleagues,

We're using a JEOL JEM 1011 TEM in our Histology Dept. since Jan. 2005. The
microscope and its ancillary devices (rotary pump, chiller, UPS etc) were on

for 24 hours almost all this time. My question is: how frequently we should
change the rotary pump oil and also which maintenances do you recommend we
should performed periodically?
Best regards.

Dr. Necat Yilmaz
Mersin University
School of Medicine
Histology & Embryology Dept.


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From: kjmorris-at-well.ox.ac.uk
Date: Tue, 30 Mar 2010 05:51:59 -0500
Subject: [Microscopy] viaWWW: Uranyl Glass

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

Hi Alex,

As well as providing a fluorescence 'standard', I believe that uranyl glass
slides could also be used to background correct uneven Hg lamp illumination,
in a similar manner as done with transmitted light illumination, as their
uV-excitation fluorescence was so uniform. I always wanted a uranium glass
slide, having worked so long on the health effects of uranium and it's
unpleasant cousins in the actinide series [Pu-238 & Pu-239] and as they just
seemed cool [they literally look brilliant in the right light, e.g. uV from
the sun or microscope] - and they are useful as that fluorescence standard
under the microscope. You can accurately measure the concentration of
uranium in solution using it's fluorescence [although I used 'delayed
neutron analysis' for uranium in tissue]. However now that the health
effects of uranium are better understood, you seldom see things made from
this material on the scientific supermarket shelves any more - no doubt
improved health and safety makes it a pain to handle for non-military
applications and buyers are now wary of uranium's radioactive and toxic
properties.

Likewise their use as a bright yellow colourant in pottery glazes and glass
has largely been abandoned, and predictably it's made them quite collectable
as secondhand items as owning the final product is generally safer and far
easier than producing it [not that I fancy eating off 1930s radioactive
'FiestaWare red' dining plates, although I'm sure it's all 'safely
contained' in the glaze]. Uranium glass may just contain the uranium oxide,
producing a clear yellow/yellow-green glass, or other minerals may be added
to produce an opalescent white [Vaseline] glass - and these household items
can be easily bought on eBay.

I have never managed to find a supplier of uranium glass 3x1" slides in the
UK, although there was a supplier of the sheet glass in the US: Newport
Industrial Glass, Inc., 1631 Monrovia Ave., Costa Mesa, CA 92627, but this
was from a listerver posting by George McNamara way back in 1995 [see
below], the company still exists though. I never got around to contacting
them. I have used suspensions of ultrafine 0.044um fluorescent fluoroscein
microspheres in solutions and gels as microscopy standards [Duke Scientific
now Thermo Scientific, also see Polysciences Inc] and I had some coloured
plastic 3x1" slides back at UCL that autofluoresced quite well [no idea
where they came from].

Used as a microscopy standard:
http://jcm.asm.org/cgi/reprint/30/5/1294.pdf
http://jcm.asm.org/cgi/reprint/27/3/442.pdf
http://www.springerlink.com/content/v122261751832k04/fulltext.pdf

Uranium Glass in science:
http://www.sis.org.uk/bulletin/92/Brenni.pdf

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford OX3 7BN,
United Kingdom.

Telephone: +44 (0)1865 287568
Email: kjmorris-at-well.ox.ac.uk
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy


Back in 1995 from a previous similar question:
----------------------------------------------------------------------------
-----------
Reply to: RE} Uranyl Glass/FM Stds.
Hi Kris,
Uranium glass slides can be purchased from:

Newport Industrial Glass, Inc.
1631 Monrovia Ave.
Costa Mesa, CA 92627
Tel: 714-642-9980
Fax: 714-645-6800
Contact person: Bill Larsen (you can tell him I sent you).
Sold as a 6.5x6.5" sheet (you can specify 1 mm height), so you may want to
form
a "consortium" to have Newport pre-cut a sheet to slide size (nominal extra
cost, but your lab only needs one or two slides). If there is a lot of
interest, my company may start selling single slides.

As for references and the Shading Correction equation: please see my article
in
the 11/94 issue of Journal of NIH Research 6(11): 80 (usual Internet
disclaimer: yes, that is an ad from my company on the facing page). Also
look
at Jericevic et al (1989) Methods in Cell Biology 30:47-83.

MutliSpeck beads: The Molecular Probes pre-mounted slide kits should be
ideal
for DAPI and Fluorescein. I believe they were optimized for Rhodamine, but
should still work ok for Texas Red. If your problem is with mounting, Mol.
Probes now sells the beads in solution, so you can 'sprinkle' some on your
specimens. If you have a different problem with the current MultiSpeck's,
Mol.
Probes may be able to work something out for you.

Sorry, but I usually buy my reference material from Mol. Probes and don't
keep
close track of other slide manufacturers.

Sincerely,

Dr. George McNamara
Universal Imaging Corporation
George_M-at-Image1.com
--------------------------------------




-----Original Message-----
X-from: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
Sent: 27 March 2010 00:59
To: kjmorris-at-well.ox.ac.uk

It is a glass that has a uranyl salt incorporated into it. The slides are
generally yellowish if I remember correctly. I bought some years ago but not
sure where. They give off a strong fluorescent signal that if resistant to
fading so they can be used as a calibration aid. The trick is measuring the
"real" signal of the unknown without it fading.

Thomas E. Phillips, Ph.D.
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
Biological Sciences
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (voice)
573-882-0123 (fax)

-----Original Message-----
X-from: bioanalytics-at-ibilabs.com [mailto:bioanalytics-at-ibilabs.com]
Sent: Friday, March 26, 2010 6:36 PM
To: Phillips, Thomas E.

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Email: bioanalytics-at-ibilabs.com
Name: Alex Besenyo PhD

Organization: IBI Labs

Title-Subject: [Filtered] Uranyl Glass

Message: Good Day,
Recently I recieved a request for information regarding Uranyl Glass
related to fluorescence intensity.

Quote: "The average NADH fluoresence intensity of each surface area
is related to the fluoresence intensity of a fluoresence calibration
glass(uranyl) and is expressed in arbitrary units."

Can anyone shed any light on what is uranyl glass?

Thank you
Alex



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44, 22 -- From kjmorris-at-well.ox.ac.uk Tue Mar 30 05:51:58 2010
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From: bfoster-at-mme1.com
Date: Tue, 30 Mar 2010 13:03:57 -0500
Subject: [Microscopy] Re: viaWWW: Uranyl Glass

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

Dear Alex

Have you tried our FluorRef slides? They are an excellent substitute for many of the things recommended below (background correction, demonstrating light paths, settting consistent power levels for lasers, etc.). They come in four different colors, to match the fluorescence of conventional fluorochromes. You can learn more about them at MicroscopyEducation.com.

Best regards,
Barbara Foster, President and Sr. Consultant

Microscopy/Microscopy Education
7101 Royal Glen Trail, Suite A
McKinney TX 75070
P: (972)924-5310 Skype: fostermme
W: www.MicroscopyEducation.com

NEWS! Visit the NEW and IMPROVED www.MicroscopyEducation.com! Now scheduling on-site courses for the balance of 2010.


At 05:06 AM 3/30/2010, kjmorris-at-well.ox.ac.uk wrote:



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From: oshel1pe-at-cmich.edu
Date: Wed, 31 Mar 2010 08:52:16 -0500
Subject: [Microscopy] Ask-A-Microscopist PAS on LX-112 embedded tissue

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

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
==========================================================

} Date: Wed, 31 Mar 2010 06:28:17 -0700 (PDT)
} From: Georgianne Ciraolo {georgianne.ciraolo-at-cchmc.org}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
} Below is the result of your form, submitted on Wednesday, March 31,
} 2010 at 06:28:00 AM.
}
} realname - Georgianne Ciraolo
} Email - georgianne.ciraolo-at-cchmc.org
} ORGANIZATION - Cincinnati Children's Hospital
} EDUCATION - Graduate College
} LOCATION - cincinnnati, ohio
} SUBJECT_OF_QUESTION - PAS stains for epon embedded sections
} QUESTION -
} One our researchers would like to do PAS staining on LX -112
} embedded tissue. Does anyone have a reliable protocol that they
} would like to share. thanks Georgianne
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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From: kjmorris-at-well.ox.ac.uk
Date: Wed, 31 Mar 2010 12:26:09 -0500
Subject: [Microscopy] Re: viaWWW: Uranyl Glass

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

Hi Barbara,

Are these 'Fluoref slides' plastic and in four pretty colours to the eye? If
so I presume they are my "I had some [4] coloured plastic 3x1" slides back
at UCL that autofluoresced quite well [no idea where they came from]". They
must have come supplied with a microscope system at some point, and were all
in one [white I think] slide box suggesting a set. Shame your website has no
pictures of them, unless these are 'on the library' virtual shelves
somewhere.

I used submicron 0.044um microspheres at a few different concentrations in
gel to create standards to investigate the 'linearity' of the confocal
system rather than background correct or use as a 'reference' fluorescence
standard, so these Fluoref standards would be useful [although they do lack
the single colour fluorescence retro charm of uranyl glass slides].

Thanks for the info.

Regards

Keith


---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford  OX3 7BN,
United Kingdom.

Telephone:  +44 (0)1865 287568
Email:  kjmorris-at-well.ox.ac.uk
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy

-----Original Message-----
X-from: bfoster-at-mme1.com [mailto:bfoster-at-mme1.com]
Sent: 30 March 2010 19:13
To: kjmorris-at-well.ox.ac.uk

Dear Alex

Have you tried our FluorRef slides? They are an excellent substitute for
many of the things recommended below (background correction, demonstrating
light paths, settting consistent power levels for lasers, etc.). They come
in four different colors, to match the fluorescence of conventional
fluorochromes. You can learn more about them at MicroscopyEducation.com.

Best regards,
Barbara Foster, President and Sr. Consultant

Microscopy/Microscopy Education
7101 Royal Glen Trail, Suite A
McKinney TX 75070
P: (972)924-5310 Skype: fostermme
W: www.MicroscopyEducation.com

NEWS! Visit the NEW and IMPROVED www.MicroscopyEducation.com! Now scheduling
on-site courses for the balance of 2010.


At 05:06 AM 3/30/2010, kjmorris-at-well.ox.ac.uk wrote:



} ---------------------------------------------------------------------------
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==============================Original Headers==============================
27, 23 -- From kjmorris-at-well.ox.ac.uk Wed Mar 31 12:26:09 2010
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From: tbogea-at-interchange.ubc.ca
Date: Wed, 31 Mar 2010 14:34:42 -0500
Subject: [Microscopy] TEM of Xenopus laevis

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

Hi everyone!

I am investigating the ultrastructure of photoreceptors in the retina of Xenopus laevis tadpoles. I've been using the TEM protocol below that was successful in the past when the eyes used to be embedded with Epon 812 resin. We are currently using EmBed 812, with DMP-30 and, although I have been able to cut semithin (~ 0.90 um thick) and ultrathin sections, the latter comes out full of holes and are very unstable under the beam. I believe this is mostly due to an infiltration problem as the ultrastructure of the tissue seemed well preserved. I am open for suggestions and will like to hear about your experience using EmBed 812 with the BDMA accelerator, which the manufacturer claims it allows for better penetration and stability. Many thanks!!

Tami


==============================Original Headers==============================
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4, 27 -- From: Tami Bogea {tbogea-at-interchange.ubc.ca}
4, 27 -- Subject: TEM of Xenopus laevis
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From: tbogea-at-interchange.ubc.ca
Date: Wed, 31 Mar 2010 14:40:56 -0500
Subject: [Microscopy] Fwd: TEM of Xenopus laevis

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

Here is the protocol:

1. Aldehyde Fix: 1% GTA/ 4% PF in 0.1M phosphate buffer (ph 7.5), cold, 48 h

2. Wash: Same buffer plus 2 drops of 1M magnesium chloride for each 20 ml of buffer, cold, at least 3 changes, 20 minutes each (Total time: 1 h to overnight)

3. Osmium fix: 1% OsO4/ 0.8% potassium ferricyanide in 0.1M phosphate buffer (ph 7.4), cold, 1 h

4. Wash: Same buffer plus 2 drops of 1M magnesium chloride for each 20 ml of buffer, cold, at least 3 changes, 20 minutes each (Total time: 1 h to overnight)

5. En bloc staining: 1% aqueous uranyl acetate, cold, 1 h

6. Dehydration: Wash 2-3x in cold distilled water, 30, 50, 70% ETOH, cold, 15-20min each, 95% and 100% (2x) ETOH, propylene oxide (in perforated caps) (2x), RT, 15-20min each

7. Infiltration (in rotator): 1 p.o.: 1 resin mixture + DMP-30), RT, 1h; 1 propylene oxide: 2 resin mix + DMP-30 (overnight) , 100% resin, RT, 30 min-2h

8. Embedding: Incubator -at- 60C, 24 h, in oven

Tami




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

} Date: Wed Mar 31 12:34:10 PDT 2010
} From: "Tami Bogea" {tbogea-at-interchange.ubc.ca}
} Subject: TEM of Xenopus laevis
} To: Microscopy-at-microscopy.com
}
} Hi everyone!
}
} I am investigating the ultrastructure of photoreceptors in the retina of Xenopus laevis tadpoles. I've been using the TEM protocol below that was successful in the past when the eyes used to be embedded with Epon 812 resin. We are currently using EmBed 812, with DMP-30 and, although I have been able to cut semithin (~ 0.90 um thick) and ultrathin sections, the latter comes out full of holes and are very unstable under the beam. I believe this is mostly due to an infiltration problem as the ultrastructure of the tissue seemed well preserved. I am open for suggestions and will like to hear about your experience using EmBed 812 with the BDMA accelerator, which the manufacturer claims it allows for better penetration and stability. Many thanks!!
}
} Tami
--
Dr. Tami Bogea
Research Associate
Ophthalmology & Visual Sciences
University of British Columbia
2550 Willow St. Vancouver, BC
V5Z 3N9 Canada
Tel: 604-875-4648
Fax: 604-875-4663


==============================Original Headers==============================
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From: kraftpiano-at-gmail.com
Date: Thu, 1 Apr 2010 09:31:57 -0500
Subject: [Microscopy] EDS: Potential application question

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

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X-from: doc.vrdoljak-at-gmail.com

Dear Listers,
We will post more detailed info when our April newsletter comes out in a
couple of weeks. Hope you can help us celebrate our 27th Symposium!

Ken Converse
NESM
kenconverse-at-qualityimages.biz

-----Original Message-----
X-from: warren moberlychan [mailto:moberlychan-at-yahoo.com]
Sent: Thursday, April 01, 2010 1:04 AM
To: Warren MoberlyChan

I was talking to a friend of mine the other day, and we were thinking that
it might be possible to get a very rough age estimate of some mineral
formations using EDS and some methods such as Rb/Sr dating. We think it
might be possible to measure the relative ratios of these two elements in a
sample (At least on a polished surface of a mineral) and get a rough
estimate of a date easily before doing more detailed analysis.

I was just wondering if anyone has done something similar, or if anyone
knows if it even can be done. We recognize that there would be a number of
assumptions made about the sample, such as uniform consistency, etc..., but
we think it might be possible. Any thoughts from the group?

--Justin.

--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar

==============================Original Headers==============================
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4, 30 -- Subject: EDS: Potential application question
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From: klivi-at-jhu.edu
Date: Thu, 1 Apr 2010 11:40:53 -0500
Subject: [Microscopy] Re: EDS: Potential application question

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

Dear Justin,
The only reliable chemical dating method uses the U/Th/Pb systematics.
Usually this is done with the mineral monazite. You have to very
accurately measure the concentrations of these elements. The
assumption is that all the Pb in the mineral comes from the decay of
Th and U. There will not be a lot of Pb depending upon the initial
concentration of Th and U and the age of the mineral. The method
work well on very old minerals (many hundreds of million years) and
using wavelength-dispersive spectroscopy. I don't think you would get
a very accurate age using EDS. Go to Mike Jercinovic's home page to
see what's being done: http://www.geo.umass.edu/faculty/jercinovic/

Ken

On Apr 1, 2010, at 10:34 AM, kraftpiano-at-gmail.com wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} I was talking to a friend of mine the other day, and we were
} thinking that
} it might be possible to get a very rough age estimate of some mineral
} formations using EDS and some methods such as Rb/Sr dating. We
} think it
} might be possible to measure the relative ratios of these two
} elements in a
} sample (At least on a polished surface of a mineral) and get a rough
} estimate of a date easily before doing more detailed analysis.
}
} I was just wondering if anyone has done something similar, or if
} anyone
} knows if it even can be done. We recognize that there would be a
} number of
} assumptions made about the sample, such as uniform consistency,
} etc..., but
} we think it might be possible. Any thoughts from the group?
}
} --Justin.
}
} --
} "America believes in education; the average professor earns more money
} in a year than a professional athlete earns in a whole week." Evan
} Esar
}
} ==============================Original
} Headers==============================
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} 4, 30 -- Subject: EDS: Potential application question
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} 4, 30 -- Content-Type: text/plain; charset=ISO-8859-1
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==============================Original Headers==============================
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From: arnec-at-bio.umass.edu
Date: Thu, 1 Apr 2010 12:05:01 -0500
Subject: [Microscopy] Nikon D5000 with fluorescence microscopy

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

Dear Listeners,

Does anybody have experience using a Nikon D5000 camera with a Nikon
fluorescent microscope? We are failing to detect signal with the green
filter set in live view mode, has anybody had a similar problem?

Thank you in advance.

Arne

==============================Original Headers==============================
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From: mcauliff-at-umdnj.edu
Date: Thu, 1 Apr 2010 12:51:48 -0500
Subject: [Microscopy] Re: Nikon D5000 with fluorescence microscopy

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

Call Nikon USA and see what they think.

Geoff

arnec-at-bio.umass.edu wrote:
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
}
} Dear Listeners,
}
} Does anybody have experience using a Nikon D5000 camera with a Nikon
} fluorescent microscope? We are failing to detect signal with the green
} filter set in live view mode, has anybody had a similar problem?
}
} Thank you in advance.
}
} Arne
}
} ==============================Original Headers==============================
} 4, 21 -- From arnec-at-bio.umass.edu Thu Apr 1 12:05:01 2010
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}
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--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff-at-umdnj.edu
**********************************************



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From: dcromey-at-email.arizona.edu
Date: Thu, 1 Apr 2010 13:33:09 -0500
Subject: [Microscopy] Re: Nikon D5000 with fluorescence microscopy

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The Nikon D5000 is a 12.3 Mpix SLR camera using a Bayer mask (or something
similar) on a CMOS chip to create color images. If my math is correct, the
15.8mm wide chip has 4288 pixels in its largest picture mode, making for (at
best) 3.7um pixels.

(Resource:
http://www.nikonusa.com/Find-Your-Nikon/Product/Digital-SLR/25452/D5000.html
)

So, you have:

* small pixels which tends to higher noise, particularly at low light level.
* each pixel is covered with either a Red, Green or Blue filter
* a CMOS chip, which (historically) has meant higher noise than a comparably
sized CCD chip
* a non-cooled sensor, meaning more noise at low light levels
* a fluorescence signal which is inherently low light

This does not sound like an optimal configuration. The reason the
scientific digital cameras (used for microscopy) are so expensive is because
the vendors work hard to deal with the many physics/optics/electronics
related issues and largely they succeed. The D5000 may be a fabulous camera
for pro-sumer photographers, but it seems less than optimal for fluorescence
imaging.

My $0.02.
Doug

^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Douglas W. Cromey, M.S. - Assistant Scientific Investigator
Dept. of Cell Biology & Anatomy, University of Arizona
1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA

office: AHSC 4212 email: Cromey-at-Arizona.edu
voice: 520-626-2824 fax: 520-626-2097

http://swehsc.pharmacy.arizona.edu/exppath/
Home of: "Microscopy and Imaging Resources on the WWW"

-----Original Message-----
X-from: mcauliff-at-umdnj.edu [mailto:mcauliff-at-umdnj.edu]
Sent: Thursday, April 01, 2010 10:53 AM
To: dcromey-at-email.arizona.edu

Call Nikon USA and see what they think.

Geoff

arnec-at-bio.umass.edu wrote:
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} Dear Listeners,
}
} Does anybody have experience using a Nikon D5000 camera with a Nikon
} fluorescent microscope? We are failing to detect signal with the green
} filter set in live view mode, has anybody had a similar problem?
}
} Thank you in advance.
}
} Arne
}
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--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
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**********************************************



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From: frah0010-at-umn.edu
Date: Thu, 1 Apr 2010 16:58:10 -0500
Subject: [Microscopy] Re: EDS: Potential application question

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Justin,

It's not a bad idea, but one of the major challenges to doing Rb/Sr dating with EDS is the poor X-ray resolution of EDS.

Consider that one would be attempting to analyze trace amounts of Rb and Sr in silicate minerals, meaning there will be a huge Si K-alpha peak at 1.740 keV. The L-alpha line for Rb is at 1.694 keV (a 46-eV difference), and the L-alpha line for Sr is at 1.807 keV (a 67-eV difference).

Even the best EDS systems have resolutions no better than 120 or 130 eV. In other words, EDS couldn't resolve Rb and Sr L peaks from the Si K peak. WDS on an electron microprobe, though, can resolve X-rays on the order of 5 eV (http://probelab.geo.umn.edu/electron_microprobe.html).

An alternative, of course, is to use the K-alpha lines for Rb and Sr, which have energies of 13.395 and 14.165 keV, respectively, but one would need an accelerating voltage of 25 kV or 30 kV to effectively excite those X-rays. If the silicates in question could handle high accelerating voltages without beam damage, that might be an option.

Otherwise, one would have to instead use an electron microprobe, and even then, tiny Rb and Sr peaks in the presence of a huge Si peak might still be a challenge and require some work. The other problem is that one would not know the ratio of Sr-87 to Sr-86 without adding another technique or making an assumption about the mineral.

As Ken noted, there is also monazite dating, using U/Th/Pb, that has been done in a few electron microprobe labs, including, most notably, Mike Jercinovic's lab at UMass. It has also been done by researchers at our lab at the University of Minnesota (http://probelab.geo.umn.edu).

Best,

-----------------
Ellery Frahm
Manager & Principal Analyst, Electron Microprobe Lab
Senior Research Fellow, Department of Geology & Geophysics
University of Minnesota - Twin Cities
http://probelab.geo.umn.edu



On Apr 1, 2010, at 9:35 AM, kraftpiano-at-gmail.com wrote:

} I was talking to a friend of mine the other day, and we were thinking that
} it might be possible to get a very rough age estimate of some mineral
} formations using EDS and some methods such as Rb/Sr dating. We think it
} might be possible to measure the relative ratios of these two elements in a
} sample (At least on a polished surface of a mineral) and get a rough
} estimate of a date easily before doing more detailed analysis.
}
} I was just wondering if anyone has done something similar, or if anyone
} knows if it even can be done. We recognize that there would be a number of
} assumptions made about the sample, such as uniform consistency, etc..., but
} we think it might be possible. Any thoughts from the group?
}
} --Justin


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From: tbogea-at-interchange.ubc.ca
Date: Thu, 1 Apr 2010 18:03:51 -0500
Subject: [Microscopy] Re: TEM of Xenopus laevis

Contents Retrieved from Microscopy Listserver Archives
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Thanks for the suggestions: I will extend the infiltration time, including two changes of 100% resin.

Others also mentioned the precipitation artifact during the en bloc "aqueous" step but so far it's not been a problem.

I've been following the manufacturer's recipe for soft blocks (e.g., 20 ml EMbed-812, 22ml DDSA, 5 ml NMA, 0.82 ml DMP-30), which cut better compared to the medium and hard ones under our conditions. This recipe gives me 1.7% of accelerator per volume of resin.

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

} Date: Wed Mar 31 19:44:29 PDT 2010
} From: "E ANN ELLIS" {eann.ellis-at-att.net}
} Subject: Re: [Microscopy] Fwd: TEM of Xenopus laevis
} To: tbogea-at-interchange.ubc.ca
}
} From looking at the infiltration schedule it appears that you need to extend
} the infiltration schedule by at least two additional changes of pure epoxy
} resin. Two additional eight hour changes would be appropriate. It does not
} matter whather you use DMP-30 or BDMA as long as you use it at the rate of
} 2% accelerator per final volume of resin.
}
} You are lucky that you are not getting uranyl acetate-phosphate precipitate
} after fixation and washes with phosphate buffer.
}
} If you substitute EMBED 812 directly for Epon 812 in the old recipe you will
} have you will not have the same quality of block that you had previously
} with Epon 812. I have re-formulated resin mixtures which used EMBED 812
} which is a generic replacement. You probably need to increase the anhydride
} component in the resin mixture. If you are using Luft's A:B mixture and
} tell me what ratio you are using and I can send you a formulation that will
} not be too soft and will section more like what you previously had.
}
} E. Ann Ellis
} Sr. Res. Associate
} Microscopy and Imaging Center
} Texas A&M University
} ----- Original Message -----
} From: {tbogea-at-interchange.ubc.ca}
} To: {eann.ellis-at-worldnet.att.net}
} Sent: Wednesday, March 31, 2010 1:42 PM
} Subject: [Microscopy] Fwd: TEM of Xenopus laevis
}
}
} }
} }
} }
} } --------------------------------------------------------------------------
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} }
} } Here is the protocol:
} }
} } 1. Aldehyde Fix: 1% GTA/ 4% PF in 0.1M phosphate buffer (ph 7.5), cold, 48
} h
} }
} } 2. Wash: Same buffer plus 2 drops of 1M magnesium chloride for each 20 ml
} of buffer, cold, at least 3 changes, 20 minutes each (Total time: 1 h to
} overnight)
} }
} } 3. Osmium fix: 1% OsO4/ 0.8% potassium ferricyanide in 0.1M phosphate
} buffer (ph 7.4), cold, 1 h
} }
} } 4. Wash: Same buffer plus 2 drops of 1M magnesium chloride for each 20 ml
} of buffer, cold, at least 3 changes, 20 minutes each (Total time: 1 h to
} overnight)
} }
} } 5. En bloc staining: 1% aqueous uranyl acetate, cold, 1 h
} }
} } 6. Dehydration: Wash 2-3x in cold distilled water, 30, 50, 70% ETOH,
} cold, 15-20min each, 95% and 100% (2x) ETOH, propylene oxide (in perforated
} caps) (2x), RT, 15-20min each
} }
} } 7. Infiltration (in rotator): 1 p.o.: 1 resin mixture + DMP-30), RT, 1h;
} 1 propylene oxide: 2 resin mix + DMP-30 (overnight) , 100% resin, RT, 30
} min-2h
} }
} } 8. Embedding: Incubator -at- 60C, 24 h, in oven
} }
} } Tami
} }
} }
} }
} }
} } -----Original Message-----
} }
} } } Date: Wed Mar 31 12:34:10 PDT 2010
} } } From: "Tami Bogea" {tbogea-at-interchange.ubc.ca}
} } } Subject: TEM of Xenopus laevis
} } } To: Microscopy-at-microscopy.com
} } }
} } } Hi everyone!
} } }
} } } I am investigating the ultrastructure of photoreceptors in the retina of
} Xenopus laevis tadpoles. I've been using the TEM protocol below that was
} successful in the past when the eyes used to be embedded with Epon 812
} resin. We are currently using EmBed 812, with DMP-30 and, although I have
} been able to cut semithin (~ 0.90 um thick) and ultrathin sections, the
} latter comes out full of holes and are very unstable under the beam. I
} believe this is mostly due to an infiltration problem as the ultrastructure
} of the tissue seemed well preserved. I am open for suggestions and will like
} to hear about your experience using EmBed 812 with the BDMA accelerator,
} which the manufacturer claims it allows for better penetration and
} stability. Many thanks!!
} } }
} } } Tami
} } --
} } Dr. Tami Bogea
} } Research Associate
} } Ophthalmology & Visual Sciences
} } University of British Columbia
} } 2550 Willow St. Vancouver, BC
} } V5Z 3N9 Canada
} } Tel: 604-875-4648
} } Fax: 604-875-4663
} }
} }
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--
Dr. Tami Bogea
Research Associate
Ophthalmology & Visual Sciences
University of British Columbia
2550 Willow St. Vancouver, BC
V5Z 3N9 Canada
Tel: 604-875-4648
Fax: 604-875-4663


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From: tbogea-at-interchange.ubc.ca
Date: Thu, 1 Apr 2010 18:09:53 -0500
Subject: [Microscopy] RE: TEM of Xenopus laevis

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

Hi Hans

I tested the recipes for soft, medium, and hard blocks provided by EMS and soft blocks cut better under our conditions. I also realized that your recipe for hard blocks is slightly different from the one I have (20ml resin, 9 ml DDSA, 12 ml NMA, 0.62-0.82 ml DMP-30). Have you tried this recipe previously?

Best,

Tami


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

} Date: Wed Mar 31 23:51:25 PDT 2010
} From: j.janssen-at-nki.nl
} Subject: RE: [Microscopy] TEM of Xenopus laevis
} To: tbogea-at-interchange.ubc.ca
}
} Hello Tami,
} I have experienced the exact same problem when embedding skin samples. Indeed it is a infiltration problem. I never tried BDMA because I am not used to that. Instead I changed the proportions of the mixed chemicals. I use the mixture for hard blocks: 20 ml Embed 812, 6 ml DDSA, 14 ml NMA and 0,72 ml DMP-30. This mixture works fine now.
}
} Success, Hans.
} Hans Janssen
} The Netherlands Cancer Institute
} Amsterdam
}
} -----Original Message-----
} From: tbogea-at-interchange.ubc.ca [mailto:tbogea-at-interchange.ubc.ca]
} Sent: woensdag 31 maart 2010 21:40
} To: Hans Janssen
} Subject: [Microscopy] TEM of Xenopus laevis
}
} ----------------------------------------------------------------------------
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}
} Hi everyone!
}
} I am investigating the ultrastructure of photoreceptors in the retina of Xenopus laevis tadpoles. I've been using the TEM protocol below that was successful in the past when the eyes used to be embedded with Epon 812 resin. We are currently using EmBed 812, with DMP-30 and, although I have been able to cut semithin (~ 0.90 um thick) and ultrathin sections, the latter comes out full of holes and are very unstable under the beam. I believe this is mostly due to an infiltration problem as the ultrastructure of the tissue seemed well preserved. I am open for suggestions and will like to hear about your experience using EmBed 812 with the BDMA accelerator, which the manufacturer claims it allows for better penetration and stability. Many thanks!!
}
} Tami
}
}
} ==============================Original Headers==============================
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} 4, 27 -- From: Tami Bogea {tbogea-at-interchange.ubc.ca}
} 4, 27 -- Subject: TEM of Xenopus laevis
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Dr. Tami Bogea
Research Associate
Ophthalmology & Visual Sciences
University of British Columbia
2550 Willow St. Vancouver, BC
V5Z 3N9 Canada
Tel: 604-875-4648
Fax: 604-875-4663


==============================Original Headers==============================
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From: tbogea-at-interchange.ubc.ca
Date: Thu, 1 Apr 2010 18:19:22 -0500
Subject: [Microscopy] RE: Fwd: TEM of Xenopus laevis

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

Hi Edward

Thanks for your comments and protocols. It's been a common theme in the responses I received that the infiltration schedule is too short. As you mentioned, the eyes would profit from multiple changes in 100% resin and I'll incorporate this into the protocol.

Best regards,

Tami


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

} Date: Thu Apr 01 10:49:25 PDT 2010
} From: "Haller, Edward" {ehaller-at-health.usf.edu}
} Subject: RE: [Microscopy] Fwd: TEM of Xenopus laevis
} To: "tbogea-at-interchange.ubc.ca" {tbogea-at-interchange.ubc.ca}
}
} Hi, Tami,
}
} I've been following the listserver to see if you got any responses, and didn't see any. While I work with mammalian tissues, my experience has been that if I don't leave the tissue in the final 100% resin infiltration steps long enough I get holes in my tissue, or it doesn't infiltrate properly and I get symptoms when I cut similar to what you describe. I work with spinal cord and brain, both of which have myelin and a lot of connective tissue, and it's hard to get resin to infiltrate. I have also worked with eye. I actually infiltrate my tissue for days with the resin before polymerization. What I do is multiple changes in resin one day, at 2-3 hours per change, with the tissue on a rotator. I then refrigerate the tissue at 4 degrees C overnight in a tightly capped vial in resin. I allow the resin to come to room temperature the next day, make another change or two in fresh resin, then embed on the second day. This makes a big difference with my samples. You might want to start out conservatively by just adding a couple of resin changes the first day to see if this helps, but may find the overnight resin change will make all the difference in the world for you. The resin at 4 degrees will not harden or get too thick if you are using the proper amount of accelerator. I use 1.25mL of DMP-30 in my mix of 100mL of resin. I'll attach both my processing schedule for hard to embed tissue and my embedding mix recipe for you. Hope these help.
}
} Ed Haller
}
}
} Edward Haller, Lab Manager
} Integrative Biology Electron Microscopy Core
} SCA 110
} 4202 East Fowler Avenue
} Tampa, FL 33620
} (813)974-2676
} ehaller-at-cas.usf.edu
}
} ________________________________________
} From: tbogea-at-interchange.ubc.ca [tbogea-at-interchange.ubc.ca]
} Sent: Wednesday, March 31, 2010 3:47 PM
} To: Haller, Edward
} Subject: [Microscopy] Fwd: TEM of Xenopus laevis
}
} ----------------------------------------------------------------------------
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}
} Here is the protocol:
}
} 1. Aldehyde Fix: 1% GTA/ 4% PF in 0.1M phosphate buffer (ph 7.5), cold, 48 h
}
} 2. Wash: Same buffer plus 2 drops of 1M magnesium chloride for each 20 ml of buffer, cold, at least 3 changes, 20 minutes each (Total time: 1 h to overnight)
}
} 3. Osmium fix: 1% OsO4/ 0.8% potassium ferricyanide in 0.1M phosphate buffer (ph 7.4), cold, 1 h
}
} 4. Wash: Same buffer plus 2 drops of 1M magnesium chloride for each 20 ml of buffer, cold, at least 3 changes, 20 minutes each (Total time: 1 h to overnight)
}
} 5. En bloc staining: 1% aqueous uranyl acetate, cold, 1 h
}
} 6. Dehydration: Wash 2-3x in cold distilled water, 30, 50, 70% ETOH, cold, 15-20min each, 95% and 100% (2x) ETOH, propylene oxide (in perforated caps) (2x), RT, 15-20min each
}
} 7. Infiltration (in rotator): 1 p.o.: 1 resin mixture + DMP-30), RT, 1h; 1 propylene oxide: 2 resin mix + DMP-30 (overnight) , 100% resin, RT, 30 min-2h
}
} 8. Embedding: Incubator -at- 60C, 24 h, in oven
}
} Tami
}
}
}
}
} -----Original Message-----
}
} } Date: Wed Mar 31 12:34:10 PDT 2010
} } From: "Tami Bogea" {tbogea-at-interchange.ubc.ca}
} } Subject: TEM of Xenopus laevis
} } To: Microscopy-at-microscopy.com
} }
} } Hi everyone!
} }
} } I am investigating the ultrastructure of photoreceptors in the retina of Xenopus laevis tadpoles. I've been using the TEM protocol below that was successful in the past when the eyes used to be embedded with Epon 812 resin. We are currently using EmBed 812, with DMP-30 and, although I have been able to cut semithin (~ 0.90 um thick) and ultrathin sections, the latter comes out full of holes and are very unstable under the beam. I believe this is mostly due to an infiltration problem as the ultrastructure of the tissue seemed well preserved. I am open for suggestions and will like to hear about your experience using EmBed 812 with the BDMA accelerator, which the manufacturer claims it allows for better penetration and stability. Many thanks!!
} }
} } Tami
} --
} Dr. Tami Bogea
} Research Associate
} Ophthalmology & Visual Sciences
} University of British Columbia
} 2550 Willow St. Vancouver, BC
} V5Z 3N9 Canada
} Tel: 604-875-4648
} Fax: 604-875-4663
}
}
} ==============================Original Headers==============================
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Dr. Tami Bogea
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Ophthalmology & Visual Sciences
University of British Columbia
2550 Willow St. Vancouver, BC
V5Z 3N9 Canada
Tel: 604-875-4648
Fax: 604-875-4663



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From: edelmare-at-muohio.edu
Date: Fri, 2 Apr 2010 14:06:29 -0500
Subject: [Microscopy] Re: Nikon D5000 with fluorescence microscopy

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

We have been using D300's for fluoresence work (as well as BF, DF,
DIC, reflected, pol etc.) and they have been doing wonderfully -
including GFP. They are more cost effective than the D5000. We use
them tethered (Nikon Capture Software), direct save to computer and
on AC. Normally we use Live view to focus - but low light
fluoresence is tough to focus and eye-piece works much better. The
D300 has very low shutter slap and vibration, but it also has a
shutter delay to handle camera vibrations in worse case.

Most of the differences D5000 vs D300 are in the "regular"
photography roles not applicable to microscopy work.



On 1 Apr 2010 at 14:33, dcromey-at-email.arizona.edu wrote:

}
}
}
} ----------------------------------------------------------------------
} ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society
} of America To Subscribe/Unsubscribe --
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}
} The Nikon D5000 is a 12.3 Mpix SLR camera using a Bayer mask (or
} something similar) on a CMOS chip to create color images. If my math
} is correct, the 15.8mm wide chip has 4288 pixels in its largest
} picture mode, making for (at best) 3.7um pixels.
}
} (Resource:
} http://www.nikonusa.com/Find-Your-Nikon/Product/Digital-SLR/25452/D500
} 0.html )
}
} So, you have:
}
} * small pixels which tends to higher noise, particularly at low light
} level. * each pixel is covered with either a Red, Green or Blue filter
} * a CMOS chip, which (historically) has meant higher noise than a
} comparably sized CCD chip * a non-cooled sensor, meaning more noise at
} low light levels * a fluorescence signal which is inherently low light
}
} This does not sound like an optimal configuration. The reason the
} scientific digital cameras (used for microscopy) are so expensive is
} because the vendors work hard to deal with the many
} physics/optics/electronics related issues and largely they succeed.
} The D5000 may be a fabulous camera for pro-sumer photographers, but it
} seems less than optimal for fluorescence imaging.
}
} My $0.02.
} Doug
}
} ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
} Douglas W. Cromey, M.S. - Assistant Scientific Investigator
} Dept. of Cell Biology & Anatomy, University of Arizona
} 1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA
}
} office: AHSC 4212 email: Cromey-at-Arizona.edu
} voice: 520-626-2824 fax: 520-626-2097
}
} http://swehsc.pharmacy.arizona.edu/exppath/
} Home of: "Microscopy and Imaging Resources on the WWW"
}
} -----Original Message-----
} X-from: mcauliff-at-umdnj.edu [mailto:mcauliff-at-umdnj.edu]
} Sent: Thursday, April 01, 2010 10:53 AM
} To: dcromey-at-email.arizona.edu
} Subject: [Microscopy] Re: Nikon D5000 with fluorescence microscopy
}
}
}
}
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} Call Nikon USA and see what they think.
}
} Geoff
}
} arnec-at-bio.umass.edu wrote:
} }
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} ------ } } Dear Listeners, } } Does anybody have experience using a
} Nikon D5000 camera with a Nikon } fluorescent microscope? We are
} failing to detect signal with the green } filter set in live view
} mode, has anybody had a similar problem? } } Thank you in advance. } }
} Arne } } ==============================Original
} Headers============================== } 4, 21 -- From
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} 4, 21 -- Date: Thu, 01 Apr 2010 13:04:59 -0400 } 4, 21 -- From: Arne K
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}
}
} --
} --
} **********************************************
} Geoff McAuliffe, Ph.D.
} Neuroscience and Cell Biology
} Robert Wood Johnson Medical School
} 675 Hoes Lane, Piscataway, NJ 08854
} voice: (732)-235-4583
} mcauliff-at-umdnj.edu
} **********************************************
}
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}
} ==============================Original
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} microscopy-at-microscopy.com 7, 28 -- Subject: Re: [Microscopy] Nikon
} D5000 with fluorescence microscopy 7, 28 -- References:
} {201004011706.o31H67Ht020350-at-ns.microscopy.com} 7, 28 -- In-reply-to:
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}
} ==============================Original
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Richard E. Edelmann, Ph.D.
EXPO Editor, Microscopy and Microanalysis Supplement
Electron Microscopy Facility Director
364 Pearson Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-muohio.edu
http://www.emf.muohio.edu


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From: kraftpiano-at-gmail.com
Date: Fri, 2 Apr 2010 18:01:31 -0500
Subject: [Microscopy] SEM: HV Tank oil question.

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

I know I might be opening up a pandoras box of warnings here, but I am
in a situation where I have two extra HV tanks that are sitting around
not being used in microscopes, and I was thinking that it would be
more practical if I could remove the oil somehow and store it
appropriately so that I can access the components. One of the tanks
already has only half the oil it started with (It was like that when I
got it) so I can't use it as-is anyway.

My question is first of all, what is an appropriate method of handling
the oil- are gloves and an apron sufficient, or should I go full
haz-mat? Once I drain the tanks, what is a safe/effective method of
storing the oil? Will a standard PTFE 5-gallon paint-bucket type
container be OK, or do I need to get a neoprene bottle of some kind?
Finally, what is a good, safe method to clean up the remaining oil
from the tank and the components so that they can be handled safely?

Thanks,

Justin A. Kraft

--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar

==============================Original Headers==============================
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==============================End of - Headers==============================





From: karen_bentley-at-urmc.rochester.edu
Date: Mon, 5 Apr 2010 18:11:59 -0500
Subject: [Microscopy] viaWWW: Reichert UltraCut E replacement bulb

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Email: karen_bentley-at-urmc.rochester.edu
Name: Karen Bentley

Organization: University of Rochester Medical Center

Title-Subject: [Filtered] Reichert UltraCut E replacement bulb

Message: Dear Listers:
I hope someone out there can help us find a replacement bulb for the
transilluminator on our 25 year old Reichert-Jung
Ultramicrotome/Ultracut E. This bulb fits into the black connecting
cable which illuiminates the back of the chuck holding the epoxy
block.
The bulb is 6V and 1W (I don't know the amperage)and is approximately
0.9mm in length and around 2-2.5 in diameter.
The inside of the bulb has a ?silver coating.
It has 2 metal prongs and no manufacturer's name on it.
Does anyone out there have any spares or know where I can find a
vendor who might have this unique bulb?
I have already contacted Leica Service and they no longer carry the
bulb nor does Tek-Net Inc. in New Jersey.

Thanks for your help!
Karen Bentley
EM Core
University of Rochester Med.Ctr.

Login Host: 128.151.71.18
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7, 11 -- From: karen_bentley-at-urmc.rochester.edu (by way of MicroscopyListserver)
7, 11 -- Subject: viaWWW: Reichert UltraCut E replacement bulb
7, 11 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed"
==============================End of - Headers==============================




From: bozzola-at-siu.edu
Date: Mon, 5 Apr 2010 19:50:30 -0500
Subject: [Microscopy] Re: viaWWW: Reichert UltraCut E replacement bulb

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

} Title-Subject: [Filtered] Reichert UltraCut E replacement bulb

Karen,

Are you sure about your measurements? I think you mean cm not mm.

I am looking at a bulb that might be what you need. It is an Osram 64
225 FHD 6V 10W Halogen Bellaphot bulb. It has two metal prongs and
measures 27 mm long (tip to end of prongs) x 9 mm wide.

You may be able to find this bulb (or equivalent) on Bulbman.com. We
have only one, otherwise I would send it along.

Good luck.

John Bozzola

} Message: Dear Listers:
} I hope someone out there can help us find a replacement bulb for the
} transilluminator on our 25 year old Reichert-Jung
} Ultramicrotome/Ultracut E. This bulb fits into the black connecting
} cable which illuiminates the back of the chuck holding the epoxy
} block.
} The bulb is 6V and 1W (I don't know the amperage)and is approximately
} 0.9mm in length and around 2-2.5 in diameter.
} The inside of the bulb has a ?silver coating.
} It has 2 metal prongs and no manufacturer's name on it.
} Does anyone out there have any spares or know where I can find a
} vendor who might have this unique bulb?
} I have already contacted Leica Service and they no longer carry the
} bulb nor does Tek-Net Inc. in New Jersey.
}
} Thanks for your help!
} Karen Bentley
} EM Core
} University of Rochester Med.Ctr.


--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

==============================Original Headers==============================
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10, 19 -- Subject: Re: [Microscopy] viaWWW: Reichert UltraCut E replacement bulb
10, 19 -- From: John Bozzola {bozzola-at-siu.edu}
10, 19 -- To: MSAListserver {Microscopy-at-microscopy.com}
10, 19 -- Cc: karen_bentley-at-urmc.rochester.edu
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==============================End of - Headers==============================





From: gary-at-gaugler.com
Date: Mon, 5 Apr 2010 22:19:44 -0500
Subject: [Microscopy] viaWWW: Reichert UltraCut E replacement

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

Unlikely that it is 1W. Probably 100W.

Try:

http://www.topbulb.com/faq/2005/TB_Catalog_2005.pdf

Search for this:

osram 64 225 FHD

Many hits.

gary g.


At 05:51 PM 4/5/2010, you wrote:



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From: bozzola-at-siu.edu
Date: Mon, 5 Apr 2010 22:48:36 -0500
Subject: [Microscopy] RE: bulb for Ultracut E

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

This is the bulb that we have:

http://www.specialtyoptical.com/osram64225fhdesamicroscopehalogenlightbulb6volt10watt.aspx

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

==============================Original Headers==============================
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3, 16 -- Subject: RE: bulb for Ultracut E
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From: naomi_mccallum-at-health.qld.gov.au
Date: Mon, 5 Apr 2010 22:50:16 -0500
Subject: [Microscopy] Fwd: Re: viaWWW: Reichert UltraCut E replacement bulb

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

Hi Karen

I have just perused our UltraCut E bag of tricks and have found bulbs that you refer to. Sorry I can't send from Australia but have more clues to assist you.

5Volt (even though manual says 6V) 60mAmp
The secret code is: 7683 (this is printed on the white part)
If you "Search engine" this number and the word bulb you should be able to locate supplier.

Hope this helps
Naomi

} } } {bozzola-at-siu.edu} 6/04/2010 10:58 am } } }



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} Title-Subject: [Filtered] Reichert UltraCut E replacement bulb

Karen,

Are you sure about your measurements? I think you mean cm not mm.

I am looking at a bulb that might be what you need. It is an Osram 64
225 FHD 6V 10W Halogen Bellaphot bulb. It has two metal prongs and
measures 27 mm long (tip to end of prongs) x 9 mm wide.

You may be able to find this bulb (or equivalent) on Bulbman.com. We
have only one, otherwise I would send it along.

Good luck.

John Bozzola

} Message: Dear Listers:
} I hope someone out there can help us find a replacement bulb for the
} transilluminator on our 25 year old Reichert-Jung
} Ultramicrotome/Ultracut E. This bulb fits into the black connecting
} cable which illuiminates the back of the chuck holding the epoxy
} block.
} The bulb is 6V and 1W (I don't know the amperage)and is approximately
} 0.9mm in length and around 2-2.5 in diameter.
} The inside of the bulb has a ?silver coating.
} It has 2 metal prongs and no manufacturer's name on it.
} Does anyone out there have any spares or know where I can find a
} vendor who might have this unique bulb?
} I have already contacted Leica Service and they no longer carry the
} bulb nor does Tek-Net Inc. in New Jersey.
}
} Thanks for your help!
} Karen Bentley
} EM Core
} University of Rochester Med.Ctr.


--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

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10, 19 -- From: John Bozzola {bozzola-at-siu.edu}
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From: W.Muss-at-salk.at
Date: Tue, 6 Apr 2010 03:17:54 -0500
Subject: [Microscopy] RE: bulb for Ultracut E

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

================================================================================================================
Good morning,

dear Dr. Bozzola, dear Karen Bentley, dear all,

hope you had some peace, rest- and beautiful Easter holidays.
================================================================================================================

The bulb which is shown when clicking the link below is (most probably if such can be used for } back lighting {, see below) the bulb for specimen/knife section edge illumination (=)
(text from the German MANUAL for the ULTRACUT E: = "Unterflurbeleuchtung";
NV-Halogenlampe fuer Unterflurbeleuchtung = } Low voltage halogen lamp for backlight illumination {: OrderNo: 860019 )

for the ULTRACUT S, this } backlight illumination { has been indicated as the following sparepart:
"Halogen reflector lamp 8V 20 W for backlight illumination"

for the ULTRACUT R: text from the English Manual:
{darkfield backlight} , lamp housing on the rear of the microtome the light from it comes through the light guide though the knife support and knife block and illuminates the knife and specimen from below (back lighting),
Halogen lamp low voltage, OrderNo: 36 00 34 12 V / 200W



What KAREN REALLY NEEDS, is the very small bulb for the } specimen-block transillumination {, REALLY: 6V - 1 W,
which in the original German Manual for the ULTRACUT E reads "only" :
"NV-Glühlampe auf Sockel für Präparat-Innenbeleuchtung" Best.Nr./=Order No 701711273" .... which translated into English should be read the same as this is informed in the English manual of the

ULTRACUT S: "Specimen transillumination" OrderNo: 86 00 38 : pin socketbulb 6 V 1 W for segment arc and for
ULTRACUT R: "Transillumination possible" (all combinations of illumination): OrderNo: 86 00 38, =
Pin-socket bulb 6V/1W for segment arc

This is - as I have seen - a very tiny bulb, presumably not halogen but a "glow" bulb...

Since I have found that I don't have that typ of bulb in my sparepart box (which is relatively large) I shall ask our serviceman doing yearly the "rejuvenation" and maintaining of the Microtome ULTRACUT E (which is our working horse... besides the old REICHERT OMU-3) wether thye have such bulbs or not.

Best wishes and regards,

Wolfgang MUSS
SALZBURG, Austria

OR Dr. phil. Wolfgang Muss*
Head of EM-Lab
Institute of Pathology, SALK-LKH
(Salzburger Landeskliniken gemeinnuetzige GesmbH, Landeskrankenhaus = Fed. State Gen. Hosp.)
Muellner Hauptstrasse 48
A-5020 SALZBURG AUSTRIA/EUROPE
Web(German): http://www.pmu.ac.at
Phone work: +43+662+4482+4720
Mobile phone work: +43+662+4482-57704
Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o W.Muss")
E-Mail work: W.Muss-at-SALK.at
Mobile-phone private: ++43+676+5 369-456
E-Mail private: wij.muss-at-aon.at

*)registered in: www.researchgate.net as "Wolfgang MUSS"
(NB.: Registration (free of charges) necessary to find / see
personal profile as well as my publication-collection)
ResearchGATE is the leading professional network for scientists.
It's free of charge and designed to meet researchers' needs
More than 200,000 scientists have joined!

and/or/alternatively (same Lab, same address data):

Paracelsus Medical Private University (PMU)
Univ.-Institute of Pathology
Electron Microscopy Lab
Muellner Hauptstrasse 48
A-5020 SALZBURG, Austria/Europe


Ankuendigung namens der (Information on behalf of)
Society for Cutaneous Ultrastructure Research (SCUR)
PLEASE VISIT THE UPDATED WEBSITE of SCUR at
} http://www.scur.org {
-------------------------------------------------------------------------
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+++2010, Sep 07th - Sep 08th: 37th Ann. SCUR Meeting HELSINKI +++
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Held before the 40th Annual ESDR meeting
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====================================================================================================

} -----Ursprüngliche Nachricht-----
} Von: bozzola-at-siu.edu [mailto:bozzola-at-siu.edu]
} Gesendet: Dienstag, 06. April 2010 05:52
} An: Muß Wolfgang
} Betreff: [Microscopy] RE: bulb for Ultracut E
}
}
}
}
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}
} This is the bulb that we have:
}
} http://www.specialtyoptical.com/osram64225fhdesamicroscopehalogenlightbulb6volt10watt.aspx
}
} --
} John J. Bozzola, Ph.D., Director
} I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
} Southern Illinois University
} 750 Communications Drive
} Carbondale, IL 62901
} Phone: 618-453-3730
}
} ==============================Original Headers==============================
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From: dac-at-research.umass.edu
Date: Tue, 6 Apr 2010 06:33:23 -0500
Subject: [Microscopy] Re: viaWWW: Reichert UltraCut E replacement bulb

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

Hi Listers,


I think that most of you are describing the lamp in the rear that
illuminates from below.

I think the bulb she is describing is the small bulb that attaches to
the flexible wire and inserts into the chuck holder on the cutting arm
so to illuminate { {through} } the specimen block. It probably is
something like 1W. This came in different styles apparently; mine is
permanently fixed to the wire. I had a conversation with someone and
theirs, like Karen's, was re-lampable. But I don't know the part.


Dale



karen_bentley-at-urmc.rochester.edu wrote:
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} Email: karen_bentley-at-urmc.rochester.edu
} Name: Karen Bentley
}
} Organization: University of Rochester Medical Center
}
} Title-Subject: [Filtered] Reichert UltraCut E replacement bulb
}
} Message: Dear Listers:
} I hope someone out there can help us find a replacement bulb for the
} transilluminator on our 25 year old Reichert-Jung
} Ultramicrotome/Ultracut E. This bulb fits into the black connecting
} cable which illuiminates the back of the chuck holding the epoxy
} block.
} The bulb is 6V and 1W (I don't know the amperage)and is approximately
} 0.9mm in length and around 2-2.5 in diameter.
} The inside of the bulb has a ?silver coating.
} It has 2 metal prongs and no manufacturer's name on it.
} Does anyone out there have any spares or know where I can find a
} vendor who might have this unique bulb?
} I have already contacted Leica Service and they no longer carry the
} bulb nor does Tek-Net Inc. in New Jersey.
}
} Thanks for your help!
} Karen Bentley
} EM Core
} University of Rochester Med.Ctr.
}
} Login Host: 128.151.71.18
} ---------------------------------------------------------------------------
}
} ==============================Original Headers==============================
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==============================Original Headers==============================
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From: oshel1pe-at-cmich.edu
Date: Tue, 6 Apr 2010 13:14:43 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist

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

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
==========================================================

} Date: Tue, 6 Apr 2010 10:49:42 -0700 (PDT)
} From: "Ronald E. Gordon" {Ronald.Gordon-at-mountsinai.org}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Tuesday, April 06,
} 2010 at 10:49:36 AM.
}
} realname - Ronald E. Gordon
} Email - Ronald.Gordon-at-mountsinai.org
} ORGANIZATION - Mt Sinai School of Medicine
} EDUCATION - Graduate College
} LOCATION - New York
} SUBJECT_OF_QUESTION - # of specimens/yr/tech
} QUESTION - I would like to know what is the suggested number of
} specimens for a technician to process to complesion per year in an
} electron microscopy laboratory. I recall a publication from an MSA
} journal which suggested 250/tech. I would like to know if that is
} still accepted.
}

==============================Original Headers==============================
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From: TindallR-at-missouri.edu
Date: Tue, 6 Apr 2010 13:32:32 -0500
Subject: [Microscopy] Samples per year

Contents Retrieved from Microscopy Listserver Archives
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Hi,

Our lab processed about 620 SEM and TEM biological samples last year, with the vast majority done by two technicians. Many (100's) misc. other samples were prepared (negative stains, simple sample mounts with or without coating, particulates on coated grids, materials specimens, etc.), and many of these were done either by our four technicians or clients.

My take is that 250 samples per year with full processing is a relatively light load, but keep in mind that we do primarily microwave processing which is waaaay faster.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




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From: kenconverse-at-qualityimages.biz
Date: Tue, 6 Apr 2010 14:09:16 -0500
Subject: [Microscopy] SEM: HV Tank oil question.

Contents Retrieved from Microscopy Listserver Archives
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Justin,
When were they manufactured? PCBs were banned in the US around 1977. I
don't believe the newer dielectric oils are a problem, besides being messy.

http://www.ehso.com/pcbs.htm
http://www.campuserc.org/resources/EHSguide/TSCA/Pages/PCBs.aspx
http://www.greenfacts.org/en/pcbs/l-2/1-polychlorinated-biphenyls.htm

The above links should give you some help in terms of determining if PCBs
are an issue.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com]
Sent: Friday, April 02, 2010 7:05 PM
To: kenconverse-at-qualityimages.biz

I know I might be opening up a pandoras box of warnings here, but I am
in a situation where I have two extra HV tanks that are sitting around
not being used in microscopes, and I was thinking that it would be
more practical if I could remove the oil somehow and store it
appropriately so that I can access the components. One of the tanks
already has only half the oil it started with (It was like that when I
got it) so I can't use it as-is anyway.

My question is first of all, what is an appropriate method of handling
the oil- are gloves and an apron sufficient, or should I go full
haz-mat? Once I drain the tanks, what is a safe/effective method of
storing the oil? Will a standard PTFE 5-gallon paint-bucket type
container be OK, or do I need to get a neoprene bottle of some kind?
Finally, what is a good, safe method to clean up the remaining oil
from the tank and the components so that they can be handled safely?

Thanks,

Justin A. Kraft

--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar

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From: Edward.Calomeni-at-osumc.edu
Date: Tue, 6 Apr 2010 14:56:18 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist

Contents Retrieved from Microscopy Listserver Archives
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Ronald,

At one time the 250 specimens per year was a valid number, assuming you are
referring to embedding/sectioning/photographing each specimen. That was when
both film and paper were developed by hand (no film/print processors). With
the use of print/film processors, around 400 samples were easily performed
per year per technician. The total number of samples also is dependent on
the type of sample and what is wanted out of them. If only negative
staining, several hundreds per 3 month period is not out of the question.

I am in a clinical pathology lab, using a tissue processor at night, rapid
polymerization during the next morning, sectioning and scoping in afternoon.
On average I process 3-4 blocks per tissue, obtain thick sections on each,
then thin section only one of the blocks. The scope is outfitted with a
digital camera and digital images are sent to the pathologist for review. No
printing is done. Last year with myself and a part-time technician, we
processed just over 1000 samples.

The use of specimens/tech/year is to be used only as a ballpark statistic.
How many other duties does the technician perform? Such as: updating SOP's,
ordering, maintaining equipment, attending mandatory meetings, etc,
etc.......

Best of luck,

Ed


Edward P. Calomeni
Director EM Lab - Pathology
The Ohio State University
M018 Starling Loving Hall
320 W. 10th Ave.
Columbus, OH 43210
614-293-5580 (office)
614-293-8806 (lab)
edward.calomeni-at-osumc.edu
-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Tuesday, April 06, 2010 2:19 PM
To: Calomeni, Edward

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA, and
any reply should go directly to the poster as well as to the list.
==========================================================

} Date: Tue, 6 Apr 2010 10:49:42 -0700 (PDT)
} From: "Ronald E. Gordon" {Ronald.Gordon-at-mountsinai.org}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Tuesday, April 06, 2010
} at 10:49:36 AM.
}
} realname - Ronald E. Gordon
} Email - Ronald.Gordon-at-mountsinai.org
} ORGANIZATION - Mt Sinai School of Medicine EDUCATION - Graduate College
} LOCATION - New York SUBJECT_OF_QUESTION - # of specimens/yr/tech
} QUESTION - I would like to know what is the suggested number of
} specimens for a technician to process to complesion per year in an
} electron microscopy laboratory. I recall a publication from an MSA
} journal which suggested 250/tech. I would like to know if that is
} still accepted.
}

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From: levilr-at-ptd.net
Date: Tue, 6 Apr 2010 18:50:27 -0500
Subject: [Microscopy] viaWWW: schematic for Nikon Labophot II- 2S001-034-1 board

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Email: levilr-at-ptd.net
Name: Lee Levine

Organization: Process Solutions Consulting

Title-Subject: [Filtered] schematic for Nikon Labophot II- 2S001-034-1 board

Message: Does anyone have a schematic for the 2S001-034-1 board? It's
the board for the 6V/20W halogen light source.

Thanks

Login Host: 70.44.185.34
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7, 11 -- Subject: viaWWW: schematic for Nikon Labophot II- 2S001-034-1 board
7, 11 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed"
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From: janec2003-at-gmail.com
Date: Tue, 6 Apr 2010 19:12:17 -0500
Subject: [Microscopy] viaWWW: a research associate position case western reserve

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

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Email: janec2003-at-gmail.com
Name: jane chen

Organization: bradley university

Title-Subject: [Filtered] a research associate position

Message: There is two intermediate research associate openings in
materials science and engineering department at case western reserve
university, this position needs microscopy skills in FIB and TEM, and
background/research experience in ceramics is preferred. If you have
interest, please contact with janec2003-at-gmail.com.

Login Host: 129.22.126.138
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From: Pamela.Lloyd-at-WPAFB.AF.MIL
Date: Wed, 7 Apr 2010 06:53:12 -0500
Subject: [Microscopy] MSORV Spring Meeting

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

The spring MSORV meeting is fast approaching.....Wednesday, April 21,
2010 at the GE Learning Center in Evandale, OH from 3-7:15PM. There is
no registration fee to attend this meeting, however, I must have your
name and affiliation to put on the list for the receptionist and the
guard in the lobby of the Learning Center as well as a head count for
the mixer/reception. Details of the meeting can be found on the MSORV
website - www.msorv.org . Directions, and abstracts and bios for the
speakers can be found there. RSVP to: pamela.lloyd-at-wpafb.af.mil


MSORV Spring Meeting
GE Learning Center
April 21, 2010

Agenda:
3-4PM Mixer/Reception -- Visit with Vendors
Sponsored by JEOL USA Inc., Electron Microscopy Sciences/Diatome and
MSORV.
4-4:15PM - Welcome (Matt Chestnut, MSORV President and from one of the
managers at GE)
4:15-5:15PM - Donna Guarrera from JEOL on the Clairscope "Atmospheric
Scanning Electron Microscopy a New Correlative Microscopy Tool".
5:15-5:30PM - 15 minute break
5:30-6:00PM - Doug Pridemore, GE "Metallurgical Evaluation of
Fractured High Pressure Turbine Forward Outer Seal".
6-6:15PM Business Meeting
6:15-7:15PM Joe Michael, MAS Tour Speaker "Microanalysis and the
FBI's Amerithrax Investigation of the 2001 Anthrax Attacks".




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From: beth-at-plantbio.uga.edu
Date: Wed, 7 Apr 2010 15:44:58 -0500
Subject: [Microscopy] cameras for light microscopy

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

Hi all,
I have camera quotes from Zeiss (AxioCam HRc Rev. 3 with Firewire or
Axiocam MRc5 High Resolution Camera) and Spot Imaging Solutions (RT3 2
MP Slider or Xplorer 1.4 MP Slider camera) for my Zeiss Axioskop2
light microscope. If you own any of these cameras I would appreciate
hearing from you.
Or, if you prefer another company I would appreciate knowing that
info, too. You can email me off-list.
thanks!
Beth



**********************************************************************
Beth Richardson
Electron Microscopy Lab Coordinator
Plant Biology Department
Miller Plant Sciences Bldg
120 Carlton Street
University of Georgia
Athens, GA 30602-7271

http://www.plantbio.uga.edu/emlab/

Phone - (706) 542-1790 & FAX - (706) 542-1805

"Between the two evils,
I always pick the one I never tried before". Mae West (1893-1980)
*******************************************************************

"And it's only the giving that makes you what you are".
Wond'ring Aloud, Jethro Tull (Aqualung)

***************************************************************************
The Friends of the Marine Institute - Join Today!
www.friendsofugami.com






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From: j.nailon-at-uq.edu.au
Date: Thu, 8 Apr 2010 00:42:49 -0500
Subject: [Microscopy] Detector Housing

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

Does anyone out there have an unwanted Secondary Electron Detector
housing from a JEOL JSM-880 or 890 SEM.
This housing could be moved in and out to adjust the position of the
PMT.
Thanks
John

John V Nailon
Operations Manager
Centre for Microscopy and Microanalysis
AIBN Building (75)
University of Queensland
St.Lucia QLD 4072
Phone: 07 3346 3988
Fax: 07 3346n 3993
email: J.Nailon-at-uq.edu.au



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From: ech-at-uvic.ca
Date: Fri, 9 Apr 2010 17:52:24 -0500
Subject: [Microscopy] viaWWW: who did these images

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

Dear listers,
The registration deadline for our 27th Annual Woods Hole Symposium has been
moved up to April 15. Our April newsletter can be found at
http://nesm.cims.harvard.edu/

I am forwarding the following message from our President, Warren
MoberlyChan, in the hopes that some of you will join us and we can get to
know you.

Ken Converse
President-Elect
NESM

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: warren moberlychan [mailto:moberlychan-at-yahoo.com]
Sent: Wednesday, April 07, 2010 9:54 PM
To: Warren MoberlyChan

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at
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Email: ech-at-uvic.ca
Name: Dr. Elaine Humphrey

Organization: University of Victoria

Title-Subject: [Filtered] who did these images

Message: Can anyone tell me who did these clever images?
http://www.dailymotion.com/video/x4mukm_zoom-into-plastic_tech

Thanks
Elaine
STEHM Technologist and Lab Manager
Advanced Microscopy Facility


Login Host: 142.104.215.62
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From: eschumacher-at-mccrone.com
Date: Mon, 12 Apr 2010 09:28:52 -0500
Subject: [Microscopy] Meeting Announcement: Midwest Microscopy and Microanalysis Society

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

X-from: Don Chernoff at ASM
To: ech-at-uvic.ca
Cc: Microscopy List
Sent: Sunday, April 11, 2010 10:45 PM

Greetings All,

The Midwest Microscopy and Microanalysis Society will meet on Friday, April 30th, 2010 for "M3S Hosts M3B: Microscopy and Microanalysis for Materials and Biology". The meeting will be held at the University of Wisconsin, Madison, and an excellent program has been arranged, including a morning plenary session followed by parallel afternoon sessions for physical and biological sciences. Details and registration information can be found on our website under Meetings:

www.midwestmicroscopy.org

When sending your RSVP, please be sure to indicate your lunch selection. We look forward to seeing you in Madison!

Elaine Schumacher
M3S Program Coordinator


*********************************************************************
Elaine F.Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com



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intended recipient. If you are not the intended recipient,
disclosure, copying, use or distribution of the information
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From: frah0010-at-umn.edu
Date: Mon, 12 Apr 2010 14:48:30 -0500
Subject: [Microscopy] BPA and mounting epoxies

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

Fellow microscopists,

Three years ago or so, when I last ordered two-part mounting epoxy for our lab and scrutinized the MSDS, I wouldn't have given a second thought to the fact that the main ingredient of the part is Bisphenol A, more commonly known as BPA.

Since then, government and scientific reports have questioned its safety, a FDA report this year reinforced concerns about health effects, and companies are eliminating BPA from their products, especially those for babies, children, and pregnant women.

Now it is time to order more mounting epoxy for our lab, and this time, upon reviewing the MSDS, BPA as a major ingredient raised a red flag.

How have others dealt with this problem? Have you found epoxies free of BPA? Has anyone switched to, say, acrylic instead? Something else entirely? Should we even be concerned about BPA in mounting media? Are the alternatives just as potentially harmful?

Any info about the safety of BPA in mounting media, alternatives to BPA-based epoxies, and other suggestions are welcome.

Best,
Ellery

-----------------
Ellery Frahm
Manager & Principal Analyst, Electron Microprobe Lab
Senior Research Fellow, Department of Geology & Geophysics
University of Minnesota - Twin Cities
http://probelab.geo.umn.edu



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From: r-holdford-at-ti.com
Date: Mon, 12 Apr 2010 20:57:41 -0500
Subject: [Microscopy] Re: viaWWW: who did these images

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

According to http://www.dailymotion.com/Weird_Weird_Science, his name is
John Sizemore and he lives in L.A.

On 4/9/10 5:52 PM, ech-at-uvic.ca wrote:
} ----------------------------------------------------------------------------
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} Email: ech-at-uvic.ca
} Name: Dr. Elaine Humphrey
}
} Organization: University of Victoria
}
} Title-Subject: [Filtered] who did these images
}
} Message: Can anyone tell me who did these clever images?
} http://www.dailymotion.com/video/x4mukm_zoom-into-plastic_tech
}
} Thanks
} Elaine
} STEHM Technologist and Lab Manager
} Advanced Microscopy Facility
}
}
} Login Host: 142.104.215.62
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} ==============================Original Headers==============================
} 8, 11 -- From zaluzec-at-microscopy.com Fri Apr 9 17:52:23 2010
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--
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Becky Holdford (r-holdford-at-ti.com)
972-995-2360
972-648-8743 (pager)
SC Packaging FA
Texas Instruments, Inc.
13536 N. Central Expressway MS 940
Dallas, TX 75243
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~


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From: nizets2-at-yahoo.com
Date: Tue, 13 Apr 2010 04:05:09 -0500
Subject: [Microscopy] BPA and mounting epoxies

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

Hi Ellery!

We are coping every day with toxic/dangerous stuff. As professionals, we are supposed to handle these products accordingly and under these conditions I don't consider myself (and my colleagues) in danger. I have always been said that the components of epoxies are made inert after polymerisation, so the trick is to work with gloves under a hood and to avoid adding the components in your coffee or licking the bottles. I am barely joking. I am pretty sure that avoiding any contact with your body should keep it safe. Even the polymerized form, you cannot say that it is over-manipulated.
I am always shocked to see that we have to store ethanol and methanol with toxic substances and work with them under the hood, when 96% alcohol bottles are available in stores and that tobacco products are freely available all over the world. As I already pointed out before, eating red meat a life long (which is directly related to colorectal cancer) or smoking are probably far more dangerous than handling these kind of stuff properly a life long.

In conclusion, I would say that yes you can search for alternatives if they exist. But don't let the MSDS over-concern you (personally I use them more as a guide for correct handling rather than a warning for just handling dangerous stuff).

Best regards,

Stephane


 


----- Original Message ----
X-from: "frah0010-at-umn.edu" {frah0010-at-umn.edu}
To: nizets2-at-yahoo.com
Sent: Mon, April 12, 2010 9:54:13 PM

Fellow microscopists,

Three years ago or so, when I last ordered two-part mounting epoxy for our lab and scrutinized the MSDS, I wouldn't have given a second thought to the fact that the main ingredient of the part is Bisphenol A, more commonly known as BPA. 

Since then, government and scientific reports have questioned its safety, a FDA report this year reinforced concerns about health effects, and companies are eliminating BPA from their products, especially those for babies, children, and pregnant women.

Now it is time to order more mounting epoxy for our lab, and this time, upon reviewing the MSDS, BPA as a major ingredient raised a red flag.

How have others dealt with this problem?  Have you found epoxies free of BPA?  Has anyone switched to, say, acrylic instead?  Something else entirely?  Should we even be concerned about BPA in mounting media?  Are the alternatives just as potentially harmful?

Any info about the safety of BPA in mounting media, alternatives to BPA-based epoxies, and other suggestions are welcome.

Best,
Ellery

-----------------
Ellery Frahm
Manager & Principal Analyst, Electron Microprobe Lab
Senior Research Fellow, Department of Geology & Geophysics
University of Minnesota - Twin Cities
http://probelab.geo.umn.edu



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From: RRA-at-stowers.org
Date: Tue, 13 Apr 2010 10:55:05 -0500
Subject: [Microscopy] FW: This is a test

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

Dear All,

The Central States Microscopy and Microanalysis Society Society will hold its Spring Meeting, "The Microscopy Toolbox", at the Stowers Institute in Kansas City, MO on Thursday and Friday, April 29-30. This excellent program and venue will include a tour of imaging and microscopy labs at Stowers and workshops the first day along with a wide variety of speakers on Friday.

For the CASINO and DTSA workshops, attendees will need to bring their own laptops. Space is limited for all 3 workshops. RSVP Rhonda Trimble at rra-at-stowers.org to preregister for the meetings and workshops. The schedule is below and more information can be found at www.emc.missouri.edu/csmms/index.html.

Hope top see you there!
Rhonda Trimble
----------------------
Central Stated Microscopy and Microanalysis Society's 2010 Spring Meeting "The Microscopy Toolbox"

Stowers Institute 1000 E. 50th St. * Kansas City, MO 64110

Thursday, April 29, 2010
9:00 - 10:00 Registration and Breakfast
10:00 - 10:15 Welcome: Rhonda Trimble, Stowers Institute
10:15 - 10:30 Opening Remarks: Dr. Winfried Wiegraebe, Director of Microscopy Center, Stowers Institute
10:30 - 12:00 Tour of Stowers imaging and analytical facilities
10:30 - 12:00 Workshop A: "CASINO" Paul Carpenter, Washington university, St. Louis, MO
12:00 - 1:15 Lunch
1:30 - 4:30 Workshop B: "A Practical Introduction To DTSA-II" Nicholas Ritchie, NIST, Gaithersburg, MO
1:30 - 4:30 Workshop C: "Thawed Cryosections for Immunocytochemistry". Paul Webster, House Ear Institute, LA, CA
6:00 PM Dinner: Jack's Stack BBQ

Friday, April 30, 2010
7:45 - 8:25 Registration and Breakfast
8:25 - 8:30 Welcome: Lou Ross, University of Missouri
8:30 - 9:15 "An Introduction to Image J" - Richard Alexander, Stowers Institute
9:15 - 10:00 "Modeling Secondary Fluorescence in Inclusions and Interfaces using DTSA-II" - Nicholas Ritchie, NIST
10:00 - 10:15 Break
10:15 - 11:00 "Utilizing TEM to understand how the obesity epidemic is causing chronic kidney disease in "couch potato"
adolescents." - M. R. Pete Hayden, University of Missouri
11:00 - 12:00 "Biofilms: another way of looking at Bacteria" - Paul Webster, House Ear Institute, Los Angeles, CA
12:00 - 12:15 Group photo, B1 Gallery Stairs
12:15 - 1:30 Lunch
1:30 - 2:00 "Moving forward without the motor: how yeast cells perform cytokinesis in absence of Myosin-II" - Giulia
Rancati, Stowers Institute
2:00 - 2:30 "Electron-probe Microanalysis: Issues for Analysis of Biological and Beam Sensitive Materials" - Paul
Carpenter, Washington University
2:30 - 2:45 Break
2:45 - 3:45 Microscopy Society of America Tour Speaker KEYNOTE ADDRESS: "Tissue engineering of models of human digits
and ears", William Landis, University of Akron, OH
3:45 - 3:50 Concluding remarks: Paul Carpenter, CSMMS President
3:50 - 4:30 CSMMS Business Meeting
4:30 Meeting adjourned

Lodging
The Stowers Institute has arranged a rate of $64/night at the Homestead Studio Suites on the plaza:. There are 20 rooms reserved at this rate. For more information go to: http://www.homesteadhotels.com/hotels/kansas-city-country-club-plaza.html.






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From: ph2-at-sprynet.com
Date: Tue, 13 Apr 2010 13:12:16 -0500
Subject: [Microscopy] BPA and mounting epoxies

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

Stephan

I can understand your comments about MSDS sheets particularly when
some companies seem to bombard you with several pages of information.
However I do find that other sources tend to give very succinct and
useful information.

I would however say that I'm not happy about grouping methanol with
ethanol. Ethanol may be recreational in appropriate quantities but
methanol certainly isn't and toxic is quite accurate.

Malcolm


Malcolm Haswell
Electron Microscope Unit
Faculty of Applied Sciences
University of Sunderland
SUNDERLAND
SR1 3SD
UK

email: malcolm.haswell-at-sunderland.ac.uk


----- Original Message -----
X-from: nizets2-at-yahoo.com

The American Industrial Hygiene Association (AIHA) is currently working on
an occupational exposure limit (Workplace Environmental Exposure Limit
(WEEL)) for BisPhenol A. [Disclosure: I'm a past chair of the committee and
current publications coordinator]. It probably won't be ready for a nother
year or two. However, let me put out some background on Bis-A (BPA)

- you can skip to the bottom if you want to bypass the supporting info and
get to the short answer.


Bisphenol A has a negligible vapor pressure and thus presents no significant
vapor hazard. It appears to have generally low skin and eye irritancy, has
a low order of acute toxicity by the oral, dermal, and inhalation routes,
and is a weak respiratory irritant. BPA is not genotoxic and a single large
dose appears to be almost totally eliminated from the rat within 48 hours.

X-from a subchronic inhalation study, 10 mg/m3 was a LOEL
(Lowest-Observed-Effect Level)that was close to a NOEL (No-Observed-Effect
Level), with signs of slight respiratory irritation in some animals and a
slight body weight effect (~3% decrease from controls) that is likely not
biologically significant. There were no histopathologic lesions reported in
these animals. Even at the next higher dose (50 mg/m3) the body weight
effects were still minimal (~5% decrease from controls) and the
histopathological lesions were judged to be slight and were limited to the
anterior nasal cavity.

Subchronic toxicity studies (all ~13 weeks) with exposure to BPA in the diet
indicated the NOEL in rats was approximately 25 mg/kg; the NOEL in mice was
{500 mg/kg based on histological findings; and the NOEL in dogs was 75
mg/kg.

The oncogenicity study in rats and mice indicated that BPA was not
carcinogenic but the NOEL in mice was {143 mg/kg based on BW effects; the
NOEL in rats was {50 mg/kg based on body weight effects.

Of the several studies performed to evaluate the developmental and
reproductive toxicity of BPA, none has demonstrated any unique effects on
these parameters. Despite using some very extensive and complicated
protocols (3-generation), only with relatively high doses causing
generalized toxicity are toxic effects observed on the fetus or sex organs.

Based on all the feeding and gavage studies on BPA, it is difficult to
define where the true NOEL (No-Observed-Effect Level) is since so many
studies used very high doses, which obviously exceeded the NOAELs
(No-Observed-Adverse-Effect Level)for rats and mice. The subchronic feeding
studies in rats indicated that the NOAEL was approximately 25 mg/kg while
the 3-generation study (lowest NOAEL in a reproduction study with a dietary
exposure) with a similar dosing duration indicated a NOEL of 5 mg/kg. The
subchronic NOEL in dogs is 75 mg/kg. These data would support a workplace
limit of } 5 mg/m3. The subchronic inhalation study appears to be the most
appropriate and sensitive study on which to base a WEEL. Minor effects
occurred at 10 mg/m3, with fairly minimal changes occurring at the next
higher concentration (50 mg/m3). From old reports in human workers,
concentrations up to 15 mg/m3 have not been found to be irritating in one
report 5 mg/m3 was noted to be an irritating concentration.

Because of the number of reports indicating that BPA is a potential skin
sensitizer in humans, albeit likely a weak one, this compound should also
have a notation indicating that it is a dermal sensitizer.

BPA has been shown to possess weak estrogenic activity (i.e., endocrine
disruptor, estrogen mimic, xenoestrogen) in probably } 200 "specialized"
studies. These studies generally show some effect on sex organs, hormone
levels, etc., in rodent offspring exposed in utero or in the neonatal stage.
Furthermore, these effects are apparent at ug/kg doses. This has given rise
to the speculation that these effects occur at very low doses, then
disappear at somewhat higher doses, then become apparent again at still
higher doses (a non-monotonic dose-response). However, the studies
generally have small numbers of animals/group and the relative potency of
BPA appears to be several orders of magnitude lower than the normal sex
hormones. Furthermore, when classical toxicology studies are performed at
high concentrations, similar toxic effects are not apparent. Therefore, for
adults in the workplace the current approach is to dismiss these "special"
studies and base a review on the classical toxicology studies.


Estimated Occupational Exposure Limit as seen by A Havics, CHMM, CIH, PE

1 mg/m3 to 5 mg/ m3 range

DSENS notation (Dermal Sensitizer)


......................................................................
Andrew Anthony "Tony" Havics, CHMM, CIH, PE
pH2, LLC
5250 E US 36, Suite 830
Avon, IN 46123
www.ph2llc.com

(317) 718-7020 off
(317) 718-7038 fax
(317) 409-3238 cell

90% of Risk Management is knowing where to place the decimal point...any
consultant can give you the other 10%(SM)

This message is from pH2. This message and any attachments may contain
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distributed without this statement.


-----Original Message-----
X-from: frah0010-at-umn.edu [mailto:frah0010-at-umn.edu]
Sent: Monday, April 12, 2010 3:55 PM
To: ph2-at-sprynet.com

Fellow microscopists,

Three years ago or so, when I last ordered two-part mounting epoxy for our
lab and scrutinized the MSDS, I wouldn't have given a second thought to the
fact that the main ingredient of the part is Bisphenol A, more commonly
known as BPA.

Since then, government and scientific reports have questioned its safety, a
FDA report this year reinforced concerns about health effects, and companies
are eliminating BPA from their products, especially those for babies,
children, and pregnant women.

Now it is time to order more mounting epoxy for our lab, and this time, upon
reviewing the MSDS, BPA as a major ingredient raised a red flag.

How have others dealt with this problem? Have you found epoxies free of
BPA? Has anyone switched to, say, acrylic instead? Something else
entirely? Should we even be concerned about BPA in mounting media? Are the
alternatives just as potentially harmful?

Any info about the safety of BPA in mounting media, alternatives to
BPA-based epoxies, and other suggestions are welcome.

Best,
Ellery

-----------------
Ellery Frahm
Manager & Principal Analyst, Electron Microprobe Lab
Senior Research Fellow, Department of Geology & Geophysics
University of Minnesota - Twin Cities
http://probelab.geo.umn.edu



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From: rfklie-at-uic.edu
Date: Tue, 13 Apr 2010 16:50:34 -0500
Subject: [Microscopy] Postdoc position in Z-contrast imaging at the University of

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

Post-Doctoral Research Associate – High-Resolution Analytical STEM of
Thermoelectric Oxides

A post-doctoral research associate position is available in the Nanoscale
Physics Group at the University of Illinois at Chicago to work on the
application of atomic-resolution Z-contrast imaging and electron
energy-loss spectroscopy (EELS) to thermoelectric oxide materials. This
position will be part of a larger collaboration between the University of
Illinois at Chicago (UIC) and the University of Alabama focusing on the
development of highly efficient high-temperature thermoelectric oxides.

The successful candidate will use atomic-resolution Z-contrast imaging and
electron energy-loss spectroscopy to determine the composition and
electronic structure of misfit-layered cobalt-oxides. Candidates should
have a Ph.D. in Physics, Materials Science or related disciplines. The
position requires extensive experience in transmission electron microscopy
and electron energy-loss spectroscopy. Experience working with
aberration-corrected scanning transmission electron microscopy is also
preferred.

This position provides an opportunity to work on cutting-edge research
related to alternative energy production. UIC is in the process of
purchasing a new state-of-the-art aberration-corrected cold field-emission
scanning transmission electron microscope, and the successful candidate
will be directly involved in working with this new instrument.

This postdoctoral position is available immediately, will be renewable on
an annual basis, and is anticipated for at least two years. Preference
will be given to recent PhD graduates who are currently in the US and have
a record of quality publication. Compensation will be commensurate with
qualification and experience. The position is open until it is filled.

Interested candidates can apply by email or by sending a cover letter, CV
with a complete list of publications, and contact information of 3
professional references to:

Professor Robert F Klie
Department of Physics
University of Illinois at Chicago
845 W Taylor Street, M/C 273
Chicago, IL 60607

email: rfklie-at-uic.edu

--
Robert F. Klie, PhD
Assistant Professor
University of Illinois at Chicago
Department of Physics
Chicago, IL 60607
Tel: 312-996-6064
Fax: 312-996-9016


==============================Original Headers==============================
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10, 23 -- Date: Tue, 13 Apr 2010 16:50:32 -0500
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10, 23 -- Illinois - Chicago
10, 23 -- From: "Klie, Robert Friedrich" {rfklie-at-uic.edu}
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From: mbrown-at-aaechighschools.com
Date: Tue, 13 Apr 2010 18:38:14 -0500
Subject: [Microscopy] Anybody have an SEM to donate to a high =?UTF-8?Q?school=3F?=

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

We have two functional SEMs at one of our high school campuses (S200
and an S360). We are pleased with what we can accomplish with them at
the high school level. Our district is opening a new campus in the
Phoenix area in the fall of this year and is interested in obtaining a
functional SEM for that location. If you are looking to retire/dispose
of an older instrument that is still serviceable I would love to hear
from you.

-Dr. Mike Brown
Science Dept Chair
AAEC-PV



==============================Original Headers==============================
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3, 19 -- To: Microscopy-at-Microscopy.Com
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From: cmshelton-at-srggi.com
Date: Tue, 13 Apr 2010 20:57:08 -0500
Subject: [Microscopy] viaWWW: Polishing Epoxy Mounted Cross Sections

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Email: cmshelton-at-srggi.com
Name: Cathy Shelton

Organization: SRGGI

Title-Subject: [Filtered] Polishing Epoxy Mounted Cross Sections

Message: I am a new user to epoxy and will be mounting cross
sectioned samples which will require polishing after the cure is
complete. I will be using Pelco epoxy resin with fast curing epoxy
hardener. The samples are electroplated plastic parts. I hope I have
chosen the right product for my application. The cross sectioned
pieces are being prepared for SEM evaluation. An older model Buehler
grinder/polisher with water will be used for the sample prep.
In reference to respiratory health, what precautions does someone
need to take during the epoxy polishing process? Is wearing some type
of mask recommended? If so,is a disposable particulate respirator
mask sufficient?
There is an attachment which could polish the samples overnight when
no employees are in the lab. Would it be safe for me to enter the lab
without respiratory protection to turn the grinder off and remove the
samples?
Thanks in advance for any responses.
Cathy

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6, 11 -- Subject: viaWWW: Polishing Epoxy Mounted Cross Sections
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From: heather.eberhardt-at-gmail.com
Date: Tue, 13 Apr 2010 20:57:43 -0500
Subject: [Microscopy] viaWWW: Consumption rate for sputter coater targets

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Email: heather.eberhardt-at-gmail.com
Name: Heather

Organization: Forest Research Institute

Title-Subject: [Filtered] Consumption rate for sputter coater targets

Message: Hello everyone,
We are in the process of purchasing our first SEM here at my company
and I was asked approximately how often we'll need to replace the
target for the sputter coater. I work in R&D for a pharmaceutical
company, so pretty much all of our samples will need sputtering.
Does anyone have any idea how long (# hours of use) an Au target
would last for?
Thanks in advance,
Heather

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6, 11 -- From zaluzec-at-microscopy.com Tue Apr 13 20:57:43 2010
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6, 11 -- To: microscopy-at-microscopy.com
6, 11 -- From: heather.eberhardt-at-gmail.com (by way of MicroscopyListserver)
6, 11 -- Subject: viaWWW: Consumption rate for sputter coater targets
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From: ech-at-uvic.ca
Date: Tue, 13 Apr 2010 20:58:16 -0500
Subject: [Microscopy] viaWWW: Images by John Sizemore

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Email: ech-at-uvic.ca
Name: Dr. Elaine Humphrey

Organization: University of Victoria

Title-Subject: [Filtered] Images by John Sizemore

Message: Last week I asked if anyone knew who did the images on
http://www.dailymotion.com/video/x4mukm_zoom-into-plastic_tech

Becky Holdford found somewhere on the site that they were done by
John Sizemore who lives in L.A.

So I tried googling John Sizemore and the nearest I have come to is
that he was at Stanford 1990. Can anyone give me his latest contact
please?

Many thanks
Elaine

Dr. Elaine Humphrey
STEHM Technologist and Lab Manager
Advanced Microscopy Facility
University of Victoria

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10, 11 -- To: microscopy-at-microscopy.com
10, 11 -- From: ech-at-uvic.ca (by way of MicroscopyListserver)
10, 11 -- Subject: viaWWW: Images by John Sizemore
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From: vanessa.duke-at-sydney.edu.au
Date: Tue, 13 Apr 2010 20:58:57 -0500
Subject: [Microscopy] viaWWW: Postdoctoral Research Associate

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Email: vanessa.duke-at-sydney.edu.au
Name: Vanessa Duke

Organization: The University of Sydney

Title-Subject: [Filtered] Postdoctoral Research
Associate ñ Microstructural Evolution of Advanced
Aluminium Alloys

Message: The Australian Centre for Microscopy &
Microanalysis (ACMM) is the headquarters of the
Australian Microscopy and Microanalysis Research
Facility (AMMRF), a major collaboration between
government and universities. The Centre
recognises the substantial role that microscopy,
imaging and microanalysis are set to play in the
next decade, as biotechnology and nanoscience
have increasing impact on science and society.
The Centre provides leadership, innovation and
ingenuity in Australian science and engineering
research.

Located on the Universityís Camperdown campus,
our centre hosts world-class facilities, and is
renowned for its research excellence. We are also
a major node of the ARC Centre of Excellence for
Design in Light Metals, and researchers have
access to an outstanding array of nanostructural
analysis equipment, both within the Centre and at
our partner nodes.

The ACMM is seeking a Postdoctoral Research
Associate to conduct advanced microstructural
characterisations of advanced Al alloys using
microscopy and microanalysis techniques. You
will be involved in the study of the solute
partitioning during early-stage precipitation in
advanced Al alloys with addition of
micro-alloying elements. You will use advanced
experimental techniques including Transmission
Electron Microscopy (TEM), and Atom Probe
Tomography (APT) for precipitate phase
identification and composition analysis. Our
Centre attracts high-profile scientists and
research groups, so an ability to communicate and
work with others will be crucial. In addition,
you will need to have the ability to write
quality manuscripts that are suitable for
publication in peer-reviewed journals.

To succeed, you will have a strong background in
metallurgy, proven research ability and a recent
PhD in materials science, engineering, physics,
chemistry or a related discipline. An ability to
use advanced scientific instrumentation;
knowledge and experience in transmission electron
microscopy; and atom probe tomography are also
essential. Expertise using HR TEM will be highly
advantageous.

This is the perfect opportunity for a
Postdoctoral Research Associate to conduct
innovative research to develop new deep
understanding about advanced Al alloys. This
research project will be carried out in
collaboration with Central South University,
China, and contains scope for international
travel in the form of research visits to China
and attendance at international conferences
worldwide.

The position is full-time fixed term for up to
two years, subject to the completion of a
satisfactory probation period for a new
appointee. Membership of a University approved
superannuation scheme is a condition of
employment for new appointees. Some assistance
towards relocation cost and visa sponsorship may
be available for the successful appointee if
required and level of appointment will be
commensurate with experience and qualifications.

Specific enquiries can be directed to Fabrice
NoÎl on +61 2 8627 1218 or by email
fabrice.noel-at-sydney.edu.au

CLOSING DATE: 5 May 2010


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15, 15 -- Subject: viaWWW: Postdoctoral Research Associate
15, 15 -- =?iso-8859-1?Q?=F1_Microstructural_Evolution_of_Advanced?= Aluminium
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From: hanke-at-mee-inc.com
Date: Tue, 13 Apr 2010 22:25:46 -0500
Subject: [Microscopy] Re: viaWWW: Images by John Sizemore

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

There is a Hollywood camera man named John Sizemore. I do not know if this
is the same person that has posted the videos on dailymotion.com. His
profile on LinkedIn lists him as an engineer at NBC. The internet movie
database shows the daytime soap opera, Days of Our Lives, as one of his
credits.

The videos are pretty cool although lacking somewhat in technical accuracy
- at least the ones about structural materials. My 20-yr old son even
posted the link on his facebook page.
--
Larry D. Hanke, P.E.
Materials Evaluation and Engineering, Inc.
Practical Solutions Through Technology and Innovation
http://www.mee-inc.com (763) 449-8870



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} replying
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}
} Email: ech-at-uvic.ca
} Name: Dr. Elaine Humphrey
}
} Organization: University of Victoria
}
} Title-Subject: [Filtered] Images by John Sizemore
}
} Message: Last week I asked if anyone knew who did the images on
} http://www.dailymotion.com/video/x4mukm_zoom-into-plastic_tech
}
} Becky Holdford found somewhere on the site that they were done by
} John Sizemore who lives in L.A.
}
} So I tried googling John Sizemore and the nearest I have come to is
} that he was at Stanford 1990. Can anyone give me his latest contact
} please?
}
} Many thanks
} Elaine
}
} Dr. Elaine Humphrey
} STEHM Technologist and Lab Manager
} Advanced Microscopy Facility
} University of Victoria
}
} Login Host: 142.104.55.117
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} Headers==============================
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} 10, 11 -- To: microscopy-at-microscopy.com
} 10, 11 -- From: ech-at-uvic.ca (by way of MicroscopyListserver)
} 10, 11 -- Subject: viaWWW: Images by John Sizemore
} 10, 11 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed"
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}



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7, 32 -- Subject: Re: [Microscopy] viaWWW: Images by John Sizemore
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From: r-holdford-at-ti.com
Date: Tue, 13 Apr 2010 23:19:49 -0500
Subject: [Microscopy] Re: viaWWW: Polishing Epoxy Mounted Cross Sections

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

Cathy: After the epoxy cures, the biggest worry would be the dust
generated by the grinding. Keeping the sample and grinding surface wet
will eliminate the dust. I have always felt that as long as I did my
epoxy mixing and cure in a hood and kept the water running while I
sectioned, I didn't have any respiratory worries about grinding epoxy.
Our safety people are in agreement with me. I've been polishing
epoxy-mounted cross-sections for 25+years and the worst thing that's
happened to me is grinding my nails down at odd angles.
On another note: I've never been a fan of fast-cure epoxies; they tend
to have a lot of residual stress in them and tend to pull away from the
samples. You'll just have to see how it works with your samples.

On 4/13/10 8:57 PM, cmshelton-at-srggi.com wrote:
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} Email: cmshelton-at-srggi.com
} Name: Cathy Shelton
}
} Organization: SRGGI
}
} Title-Subject: [Filtered] Polishing Epoxy Mounted Cross Sections
}
} Message: I am a new user to epoxy and will be mounting cross
} sectioned samples which will require polishing after the cure is
} complete. I will be using Pelco epoxy resin with fast curing epoxy
} hardener. The samples are electroplated plastic parts. I hope I have
} chosen the right product for my application. The cross sectioned
} pieces are being prepared for SEM evaluation. An older model Buehler
} grinder/polisher with water will be used for the sample prep.
} In reference to respiratory health, what precautions does someone
} need to take during the epoxy polishing process? Is wearing some type
} of mask recommended? If so,is a disposable particulate respirator
} mask sufficient?
} There is an attachment which could polish the samples overnight when
} no employees are in the lab. Would it be safe for me to enter the lab
} without respiratory protection to turn the grinder off and remove the
} samples?
} Thanks in advance for any responses.
} Cathy
}
} Login Host: 65.15.174.90
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--
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Becky Holdford (r-holdford-at-ti.com)
972-995-2360
972-648-8743 (pager)
SC Packaging FA
Texas Instruments, Inc.
13536 N. Central Expressway MS 940
Dallas, TX 75243
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From: bozzola-at-siu.edu
Date: Wed, 14 Apr 2010 07:54:14 -0500
Subject: [Microscopy] Re: viaWWW: Consumption rate for sputter coater targets

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

Stephane

I know that you had to clarify the use of alcohols in labs, but I
wasn't for one minute suggesting that lab ethanol had a recreational
value either in our labs or anyone else's.

Thanks

Malcolm

----- Original Message -----
X-from: Stephane Nizet {nizets2-at-yahoo.com}

Hello Heather,

Sputter-coater targets are like brakes on a car: depends on how hard
(thickness of coating) and how often you use them. Also, it depends on
the thickness of the target itself. Ask the manufacturer. They usually
have a good idea.

On average, we replace our target every 2-3 years. My recommendation
is that if you coat a lot of specimens, figure a replacement every
year and a half. There is something to be said about having one or two
in reserve and ready to go when needed.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730
}
} Title-Subject: [Filtered] Consumption rate for sputter coater targets
}
} Message: Hello everyone,
} We are in the process of purchasing our first SEM here at my company
} and I was asked approximately how often we'll need to replace the
} target for the sputter coater.  I work in R&D for a pharmaceutical
} company, so pretty much all of our samples will need sputtering.
} Does anyone have any idea how long (# hours of use) an Au target
} would last for?
} Thanks in advance,
} Heather
}
}  Login Host: 69.27.233.254
} ---------------------------------------------------------------------------
}


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From: richard.ross-at-allisontransmission.com
Date: Wed, 14 Apr 2010 08:03:42 -0500
Subject: [Microscopy] Re: viaWWW: Polishing Epoxy Mounted Cross Sections

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

Cathy,

The water used in the grinding process provides several benefits. The
removed media gets entrained into the water and goes down the drain
instead of potentially into the air. Thus, there is no need for
respiratory protection The water also cools the sample, protecting the
sample from heat damage. The entrained particles do like to settle out in
the slow-flowing drains, which may eventually plug. But, that will be a
different question to post!

I would assume your final polish is a diamond paste (or similar) on a felt
cloth; no protection is needed here as well. The felt cloth essentially
holds the debris. As the felt wears out, this debris does get spun off,
but the oil/grease in the paste entrains the particles.

Occasionally a sample gets multiple facets from a poor grinding step; a
water cooled belt sander is then nice to have to flatten the sample out
and start over.

Rick





cmshelton-at-srggi.com
04/13/2010 10:15 PM
Please respond to
cmshelton-at-srggi.com


To
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Subject
[Microscopy] viaWWW: Polishing Epoxy Mounted Cross Sections









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Email: cmshelton-at-srggi.com
Name: Cathy Shelton

Organization: SRGGI

Title-Subject: [Filtered] Polishing Epoxy Mounted Cross Sections

Message: I am a new user to epoxy and will be mounting cross
sectioned samples which will require polishing after the cure is
complete. I will be using Pelco epoxy resin with fast curing epoxy
hardener. The samples are electroplated plastic parts. I hope I have
chosen the right product for my application. The cross sectioned
pieces are being prepared for SEM evaluation. An older model Buehler
grinder/polisher with water will be used for the sample prep.
In reference to respiratory health, what precautions does someone
need to take during the epoxy polishing process? Is wearing some type
of mask recommended? If so,is a disposable particulate respirator
mask sufficient?
There is an attachment which could polish the samples overnight when
no employees are in the lab. Would it be safe for me to enter the lab
without respiratory protection to turn the grinder off and remove the
samples?
Thanks in advance for any responses.
Cathy

Login Host: 65.15.174.90
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From: DusevichV-at-umkc.edu
Date: Wed, 14 Apr 2010 09:26:14 -0500
Subject: [Microscopy] RE: viaWWW: Consumption rate for sputter coater targets

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

Do not buy Au (pretty looking specimens, grainy coating). Go instead
with Au-Pd target. Longevity depends very much on type of coater,
thickness of target and frequency of use. Ballpark estimate: 0.5-3
years.

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

} -----Original Message-----
} From: heather.eberhardt-at-gmail.com [mailto:heather.eberhardt-at-gmail.com]
} Sent: Tuesday, April 13, 2010 8:58 PM
} To: Dusevich, Vladimir
} Subject: [Microscopy] viaWWW: Consumption rate for sputter coater
} targets
}
}
}
}
}
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}
} Organization: Forest Research Institute
}
} Title-Subject: [Filtered] Consumption rate for sputter coater targets
}
} Message: Hello everyone,
} We are in the process of purchasing our first SEM here at my company
} and I was asked approximately how often we'll need to replace the
} target for the sputter coater. I work in R&D for a pharmaceutical
} company, so pretty much all of our samples will need sputtering.
} Does anyone have any idea how long (# hours of use) an Au target
} would last for?
} Thanks in advance,
} Heather
}
} Login Host: 69.27.233.254
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From: yvan_lindekens-at-yahoo.com
Date: Wed, 14 Apr 2010 10:31:44 -0500
Subject: [Microscopy] 2 tungsten carbide microtome knives for Jung Model K availlable

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

Hi all,

As I'm cleaning out my microtechnique lab I have 2 tungsten carbide microtome knives for the large base sledge microtome Jung Model K availlable.

I know from experience that these, especially for the Jung K designed and developped knives, are near to impossible to get.

The knives are some 10cm * 10cm.

If someone's interested in those, please contact me of list. I'll be happy to sell them for a small fee.

Regards,

Yvan.





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From: kraftpiano-at-gmail.com
Date: Wed, 14 Apr 2010 10:42:58 -0500
Subject: [Microscopy] SEM: JEOL Service docs.

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

I am in need of an installation/adjustment document as well as a list
of adjustments and/or calibration guide for a JEOL JSM-6100. At the
very least, I would like a startup flowchart showing the correct
sequence of events leading to HV enable.

Thanks,

Justin A. Kraft


--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar

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5, 30 -- Subject: SEM: JEOL Service docs.
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From: neal.melvin-at-gmail.com
Date: Wed, 14 Apr 2010 18:14:53 -0500
Subject: [Microscopy] viaWWW: Zeiss DL-2 Documator

Contents Retrieved from Microscopy Listserver Archives
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This Question/Comment was submitted to the Microscopy Listserver
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Email: neal.melvin-at-gmail.com
Name: Neal

Organization: UTSW

Title-Subject: [Filtered] Zeiss DL-2 Documator?

Message: I am looking to buy one of these old documators... does
anyone happen to have one that they'd be willing to part with?

Login Host: 24.238.178.83
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==============================Original Headers==============================
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From: jpshield-at-uga.edu
Date: Wed, 14 Apr 2010 18:15:42 -0500
Subject: [Microscopy] viaWWW: image of George Pappas

Contents Retrieved from Microscopy Listserver Archives
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Email: jpshield-at-uga.edu
Name: John Shields

Organization: University of Georgia

Title-Subject: [Filtered] image of George Pappas

Message: Hi Everyone,
I am in need of a relatively high resolution image of Dr. George
Pappas, the MSA 2010 Distinguished Scientist Award winner.
If you happen to have an appropriate image, please send to John
Shields (johnshields59-at-gmail.com). My gmail account can handle
larger attachments than my institution.

Thank you very much in advance!
John S
M&M Proceedings Editor

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From: benada-at-biomed.cas.cz
Date: Thu, 15 Apr 2010 04:31:07 -0500
Subject: [Microscopy] CCD camera and vacuum leak

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

Hello,
we have troubles with our MegaViewII camera mounted on CM100 and vacuum leak.
The leak occurs only when camera starts to move into the column. When it is in
working position, the vacuum recovery is really fast. No leak occurs when
camera starts to move out of the column into the park position. There is a
possibility to get the new rod seals and replaced the old ones.
Please, any MegaViewII user had to solve similar problem?

Thanking in advance for any suggestions.

With regards Oldrich

--
Oldrich Benada
Institute of Microbiology
Acad. Sci. CR
Videnska 1024
CZ-142 20 Prague 4
Czech Republic

==============================Original Headers==============================
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4, 20 -- Organization: Institute of Microbiology, Acad. Sci. CR, v.v.i.
4, 20 -- To: microscopy-at-microscopy.com
4, 20 -- Subject: CCD camera and vacuum leak
4, 20 -- Date: Thu, 15 Apr 2010 11:31:00 +0200
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From: bengu-at-fen.bilkent.edu.tr
Date: Thu, 15 Apr 2010 07:19:11 -0500
Subject: [Microscopy] LCD monitor for a Carl-Zeiss EVO40

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


Hi All,

The main monitor on our SEM (CZ-EVO 40) stopped functioning. We need to
buy a new one. The original was a 17 inch, 4:3 ratio, 1280x1024 one.

Nowadays, it is hard to find a 4:3 LCD around, all are these wide screen,
16:9, 1920x 1080 monitors. Will a 16:9 with such resolution work fine or
we need to go hunt for a 4:3 ?

Thanx

Erman Bengu



=================================
Erman Bengu

Assistant Professor of Chemistry
Department of Chemistry
Bilkent University

Mailing Address:
Bilkent University,
Department of Chemistry,
06800, Bilkent, Ankara
Turkey

Office: SB #311
E-mail: bengu_AT_fen.bilkent.edu.tr
Phone (Office): +90 (312) 290-2153
(Lab1): +90 (312) 290-2663
(Lab2): +90 (312) 290-3332
Fax: +90 (312) 266-4068
Web: http://www.fen.bilkent.edu.tr/~bengu
==================================





==============================Original Headers==============================
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16, 22 -- Date: Thu, 15 Apr 2010 15:19:36 +0300 (EEST)
16, 22 -- Subject: LCD monitor for a Carl-Zeiss EVO40
16, 22 -- From: "Erman Bengu" {bengu-at-fen.bilkent.edu.tr}
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From: John.Mardinly-at-wdc.com
Date: Thu, 15 Apr 2010 19:29:46 -0500
Subject: [Microscopy] LCD monitor for a Carl-Zeiss EVO40

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


If your video card will support the 16:9 resolution (has the appropriate
pixel array choice), the newer monitor aspect ratio should work.
However, if your card will only support 4:3 arrays, the display will
usually be stretched horizontally and distort images. It may be easier
to replace the video card and the monitor if this is the case.

Woody

N.W. (Woody) White Jr.
Electron Microscopist
Chemistry and Materials Center
AREVA NP Inc
An AREVA and Siemens Company
Lynchburg, VA
Lab Phone: 434.832.3004

-----Original Message-----
X-from: bengu-at-fen.bilkent.edu.tr [mailto:bengu-at-fen.bilkent.edu.tr]
Sent: Thursday, April 15, 2010 8:28 AM
To: WHITE Norvell (AREVA NP INC)


Hi All,

The main monitor on our SEM (CZ-EVO 40) stopped functioning. We need to
buy a new one. The original was a 17 inch, 4:3 ratio, 1280x1024 one.

Nowadays, it is hard to find a 4:3 LCD around, all are these wide
screen, 16:9, 1920x 1080 monitors. Will a 16:9 with such resolution work
fine or we need to go hunt for a 4:3 ?

Thanx

Erman Bengu



=================================
Erman Bengu

Assistant Professor of Chemistry
Department of Chemistry
Bilkent University

Mailing Address:
Bilkent University,
Department of Chemistry,
06800, Bilkent, Ankara
Turkey

Office: SB #311
E-mail: bengu_AT_fen.bilkent.edu.tr
Phone (Office): +90 (312) 290-2153
(Lab1): +90 (312) 290-2663
(Lab2): +90 (312) 290-3332
Fax: +90 (312) 266-4068
Web: http://www.fen.bilkent.edu.tr/~bengu
==================================





==============================Original
Headers==============================
16, 22 -- From bengu-at-fen.bilkent.edu.tr Thu Apr 15 07:19:11 2010 16, 22
-- Received: from ispinoz.bcc.bilkent.edu.tr (ispinoz.bcc.bilkent.edu.tr
[139.179.10.240])
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ESMTP id o3FCJBlQ010578
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07:19:11 -0500
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[192.168.10.203])
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7D61E5525
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15:20:53 +0300 (EEST)
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bengu-at-fen.bilkent.edu.tr)
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16, 22 -- Message-ID:
{55740.139.179.97.107.1271333976.squirrel-at-newmail.bilkent.edu.tr}
16, 22 -- Date: Thu, 15 Apr 2010 15:19:36 +0300 (EEST) 16, 22 --

Erman;
All of the monitors on our brand new TEM are 1280 x 1024, so
somebody is still selling obsolete monitors....... Heck, it's just a PC and
most modern cards will drive a wide variety of formats. If you don't like
the card, change it-they are cheap. In fact, you can most likely buy a 1900x
1200 monitor PLUS a top of the line video card for less than one of those
old 1200x1024 monitors, so give it a try.

John Mardinly

Western Digital



-----Original Message-----
X-from: bengu-at-fen.bilkent.edu.tr [mailto:bengu-at-fen.bilkent.edu.tr]
Sent: Thursday, April 15, 2010 5:29 AM
To: John Mardinly


Hi All,

The main monitor on our SEM (CZ-EVO 40) stopped functioning. We need to
buy a new one. The original was a 17 inch, 4:3 ratio, 1280x1024 one.

Nowadays, it is hard to find a 4:3 LCD around, all are these wide screen,
16:9, 1920x 1080 monitors. Will a 16:9 with such resolution work fine or
we need to go hunt for a 4:3 ?

Thanx

Erman Bengu



=================================
Erman Bengu

Assistant Professor of Chemistry
Department of Chemistry
Bilkent University

Mailing Address:
Bilkent University,
Department of Chemistry,
06800, Bilkent, Ankara
Turkey

Office: SB #311
E-mail: bengu_AT_fen.bilkent.edu.tr
Phone (Office): +90 (312) 290-2153
(Lab1): +90 (312) 290-2663
(Lab2): +90 (312) 290-3332
Fax: +90 (312) 266-4068
Web: http://www.fen.bilkent.edu.tr/~bengu
==================================





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16, 22 -- Subject: LCD monitor for a Carl-Zeiss EVO40
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From: benada-at-biomed.cas.cz
Date: Fri, 16 Apr 2010 05:46:29 -0500
Subject: [Microscopy] CCD camera and vacuum leak - Thanks

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

Hello,
Many thanks everyone for comments and suggestions.

My best regards from Prague
Oldrich
--
Oldrich Benada
Institute of Microbiology
Acad. Sci. CR
Videnska 1024
CZ-142 20 Prague 4
Czech Republic

==============================Original Headers==============================
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From: mmcheath-at-syr.edu
Date: Fri, 16 Apr 2010 06:03:18 -0500
Subject: [Microscopy] Available: Noran Voyager electronics

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

Hi,

I've been holding on to the electronics consoles from a Noran Voyager
EDS/Electron Microprobe control system. I have to free up space and it is
time for me to get rid of these components.

This is not a working system. It could be scavenged for spare parts.

I must get rid of it by April 30. It is free to anyone who wants to come to
Syracuse University and pick it up. The components will fit in the back of
a minivan with the rear seats removed.

Please let me know ASAP if you want it. Otherwise it is going to the
electronics recyclers.

Mike


********************************************************************
Michael M. Cheatham
312 Heroy Geology Laboratory Phone (315)-443-1261
Syracuse University Fax (315)-443-3363
Syracuse, NY 13244-1070

email:mmcheath-at-syr.edu
http://earthsciences.syr.edu

owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html
owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html
owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html
owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html
********************************************************************




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From: Frank_Karl-at-lincolnelectric.com
Date: Fri, 16 Apr 2010 06:15:38 -0500
Subject: [Microscopy] panhandling for volcanic dust

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



I was listening to National public Radio on my way to work and one of the
big stories is how the volcanic ash from Iceland is disrupting air travel
in parts of Europe.

Of course as a light microscopist I would really enjoy getting a sample of
the dust, but a trip to Iceland or England is a little out of my reach,
even for a volcanic ash sample. Then I thought of you. That's the
collective you.

If anyone finds themselves sweeping or dusting a surface and grumbling
about the darn dust, if it's not too much trouble, could you sweep a little
of it into a glassine or suitable container and mail me a sample? It
doesn't have to be too big and a few skin cells wouldn't hurt.

I'm not sure I can promise to reimburse you for the postage, but I will put
your name on the slide and sometime in the future somebody will be looking
through my estate's slide collection (ebay maybe) wondering who you are.
Think of it as a limited form of immortality.

Thanks!

Frank Karl
PC32090
Lincoln Electric Company
22801 Saint Clair Ave.
Cleveland, Ohio USA.
44117-119,


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communication in error, please notify us immediately by
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From: dsoren-at-umich.edu
Date: Fri, 16 Apr 2010 07:59:03 -0500
Subject: [Microscopy] Teorell-Stenhagen buffer

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


Hello all,

We are going to be doing some DAB staining for TEM. Several of the
protocols that I have found in the literature call for Teorell-
Stenhagen buffer, pH10.5. Does anyone have a recipe for this buffer?
The original reference dates back to 1938.

Alternately, another buffer one can use is glycine-NaOH buffer at the
same pH. Does anyone have a recipe for this one?

Thanks,

Dotty


Dorothy Sorenson
Microscopy and Image-analysis Laboratory
Department of Cell and Developmental Biology
University Of Michigan Medical School
A807 BSRB
109 Zina Pitcher Place
Ann Arbor, MI 48109-2200
(734)763-1170
FAX (734)763-1166



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From: kraftpiano-at-gmail.com
Date: Fri, 16 Apr 2010 11:59:29 -0500
Subject: [Microscopy] "Handedness" of SEMs.

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

This is not really a technical question, more just something that I've
been pondering for a little while. I've noticed through my own
experience as well as through several Google image searches that most
SEMs seem to be "Right handed," as in the controls are to the right of
the SEM column. I've noticed this across manufacturers and models. I
was wondering if anyone had an explanation as to why this is, or if it
is just random? It seems to me that it would be convenient for some
to have the controls on the left hand side.

Just curious,

Justin A. Kraft

--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar

==============================Original Headers==============================
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4, 30 -- Subject: "Handedness" of SEMs.
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From: kenconverse-at-qualityimages.biz
Date: Fri, 16 Apr 2010 12:19:14 -0500
Subject: [Microscopy] "Handedness" of SEMs.

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

Justin,
The only SEMs that I'm aware of that put the electronics on the left are
ETECs. I believe their thinking was that most people are right-handed and
would have the best control of the specimen stage using their right hand.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com]
Sent: Friday, April 16, 2010 1:06 PM
To: kenconverse-at-qualityimages.biz

This is not really a technical question, more just something that I've
been pondering for a little while. I've noticed through my own
experience as well as through several Google image searches that most
SEMs seem to be "Right handed," as in the controls are to the right of
the SEM column. I've noticed this across manufacturers and models. I
was wondering if anyone had an explanation as to why this is, or if it
is just random? It seems to me that it would be convenient for some
to have the controls on the left hand side.

Just curious,

Justin A. Kraft

--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar

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4, 30 -- Subject: "Handedness" of SEMs.
4, 30 -- From: Justin Kraft {kraftpiano-at-gmail.com}
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From: protrain-at-emcourses.com
Date: Fri, 16 Apr 2010 16:37:32 -0500
Subject: [Microscopy] "Handedness" of SEMs.

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

Hi all

Philips in the 1970s were the only major SEM manufacturer who had the column
on the right of the instrument. Once the column unit becomes self contained,
with its power supplies within the one unit, the position of the desk is a
customer option.

For example the Zeiss SEM the column has been self contained for many years
and only needs a desk for the computer on the left or right.


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com


-----Original Message-----
X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com]
Sent: 16 April 2010 18:04
To: protrain-at-emcourses.com

This is not really a technical question, more just something that I've
been pondering for a little while. I've noticed through my own
experience as well as through several Google image searches that most
SEMs seem to be "Right handed," as in the controls are to the right of
the SEM column. I've noticed this across manufacturers and models. I
was wondering if anyone had an explanation as to why this is, or if it
is just random? It seems to me that it would be convenient for some
to have the controls on the left hand side.

Just curious,

Justin A. Kraft

--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar

==============================Original Headers==============================
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4, 30 -- Subject: "Handedness" of SEMs.
4, 30 -- From: Justin Kraft {kraftpiano-at-gmail.com}
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From: raristau-at-ims.uconn.edu
Date: Fri, 16 Apr 2010 17:01:56 -0500
Subject: [Microscopy] Re: "Handedness" of SEMs.

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

This interesting observation brought several thoughts to mind (my mind being
otherwise undistracted on this slow Friday afternoon....)

If the console is on the right-hand side, then my left hand is closest to
the column for such tasks as aligning apertures and opening chamber
doors....

As I sit at the chair at the center of our ESEM console (Electroscan 2020),
the joystick for stage movements is mounted on the extreme left of the
console, again favoring left-hand manipulation....

And finally, has anyone else found that our engineering colleagues are
disproportionately left-handed? That is, a larger proportion than the
average population? Just an informal observation on my part....

Roger A. Ristau, PhD
Electron Microscopy Specialist
Institute of Materials Science
97 North Eagleville Road
University of Connecticut
Storrs, CT 06269
vox: 860-486-5453
fax: 860-486-4745


} From: {kraftpiano-at-gmail.com}
} Reply-To: {kraftpiano-at-gmail.com}
} Date: Fri, 16 Apr 2010 12:10:22 -0500
} To: {raristau-at-ims.uconn.edu}
} Subject: [Microscopy] "Handedness" of SEMs.
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} ----------------------------------------------------------------------------
}
} This is not really a technical question, more just something that I've
} been pondering for a little while. I've noticed through my own
} experience as well as through several Google image searches that most
} SEMs seem to be "Right handed," as in the controls are to the right of
} the SEM column. I've noticed this across manufacturers and models. I
} was wondering if anyone had an explanation as to why this is, or if it
} is just random? It seems to me that it would be convenient for some
} to have the controls on the left hand side.
}
} Just curious,
}
} Justin A. Kraft
}
} --
} "America believes in education; the average professor earns more money
} in a year than a professional athlete earns in a whole week." Evan
} Esar
}
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} 4, 30 -- Subject: "Handedness" of SEMs.
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} 4, 30 -- To: microscopy-at-microscopy.com
} 4, 30 -- Content-Type: text/plain; charset=ISO-8859-1
} ==============================End of - Headers==============================



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From: david.knecht-at-uconn.edu
Date: Sat, 17 Apr 2010 17:43:19 -0500
Subject: [Microscopy] Destructive Interference

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

I was teaching about phase contrast and talking about destructive interference with my class the other day and it suddenly occurred to me that I had no idea how to answer a question about the energetics of destructive interference. What happens to the energy associated with light when you have destructive interference? Also, how do you account for this phenomenon in terms of photons as opposed to waves? Dave

Dr. David Knecht
Department of Molecular and Cell Biology
Co-head Flow Cytometry and Confocal Microscopy Facility
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)




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From: Janne.Hyoetylae-at-stud.unibas.ch
Date: Sun, 18 Apr 2010 16:51:42 -0500
Subject: [Microscopy] Re: Destructive Interference

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

Dave,

How do you account for the result of the double slit interference
experiment in terms of photons? You can't, that's why there is this
particle-wave duality. In a similar way you can't account for discrete
photons in wave terms, although we have experiments that clearly show the
particle behaviour.

As for the energy, classically speaking, destructive interference just
means that the amplitude of the electric and magnetic field is zero at the
particular point in space at any time. As such, there is also no energy to
be transferred at this point. Remember that the energy comes from the
electric and magnetic field present at a point (in the wave
representation), or as discrete packets, i.e. photons (in the particle
representation). Since interference is a term related to waves, thinking
about energies should happen within the wave formalism.

Going a bit into quantum mechanics, there is a way to reconcile the two
worlds. A photon is then represented by a quantum mechanical wave
function. The shape of the function gives you a probability to detect the
photon at a given point in space and time. Destructive interference means
that at that point in space, several photon wave functions overlap in
way[*], that the probability to detect a photon there is zero. Since you
have to detect the photon somewhere in the end (i.e. the total probability
over all spaces and times is fixed), this means that there are other
points in space and/or time which "compensate" for the destructive
interference, i.e. have higher probability to detect a photon; this is
then the constructive interference.

If I was not clear enough in my explanations, don't hesitate to ask
further.

Cheers
Janne


[*] If you are still following me: even the wave function of a single
photon can overlap with itself to create interference, if the wave is
allowed to pass through separate places.


On Sun, 18 Apr 2010 00:53:24 +0200, {david.knecht-at-uconn.edu} wrote:

} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} I was teaching about phase contrast and talking about destructive
} interference with my class the other day and it suddenly occurred to me
} that I had no idea how to answer a question about the energetics of
} destructive interference. What happens to the energy associated with
} light when you have destructive interference? Also, how do you account
} for this phenomenon in terms of photons as opposed to waves? Dave
}
} Dr. David Knecht
} Department of Molecular and Cell Biology
} Co-head Flow Cytometry and Confocal Microscopy Facility
} U-3125
} 91 N. Eagleville Rd.
} University of Connecticut
} Storrs, CT 06269
} 860-486-2200
} 860-486-4331 (fax)

--
Janne Hyötylä
Biozentrum and the Swiss Nanoscience Institute
University of Basel
Klingelbergstr. 50/70
CH-4056 Basel
Switzerland

tel: +41 61 267 2082
e-mail: Janne.Hyoetylae-at-stud.unibas.ch

==============================Original Headers==============================
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From: Frank_Karl-at-lincolnelectric.com
Date: Tue, 20 Apr 2010 05:58:16 -0500
Subject: [Microscopy] Microscopy meeting in Northeastern Ohio

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


The Microscopy Society of Northeastern Ohio (MSNO) will hold its spring
dinner meeting May 6 2010 at Harry’s Steakhouse on Brecksville Rd (Rte 21)
in Independence OH.

On board with be Mike Mallamaci, Director of PolyInsight, LLC as our
keynote speaker and MSA Presidential Scholar, Karen McGuire. Both are
highly regarded and the meeting promises to quite interesting. Additional
information can be found at our website: http://www.msneo.org

Our meeting will be held at Harry’s Steakhouse on Brecksville Rd (Rte 21)
in Independence OH (www.pluggedincleveland.com/restaurants/820+harrys
+steakhouse.html ) on Thursday May 6. Appetizers starting at 4:30 and a
buffet dinner at 6:00pm will be followed by the talks.

Cost will be $25 for members, $30 for non-members, $10 student members and
$15 for student non-members. (RSVP to Pat Glazebrook at
pglazebrook-at-metrohealth.org or (216) 778-8958)

Join us for a interesting evening and sparking conversation with other
microscopist over great food. It promises to be a very interesting meeting
and a great chance to hobnob with other microscopists.


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From: kraftpiano-at-gmail.com
Date: Tue, 20 Apr 2010 10:04:37 -0500
Subject: [Microscopy] Lab (Garage) clean-out.

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

Dear listers,

I have finally finished going through all of my EM stuff that I've
collected for parts/repairs, and I have a bunch of stuff I know I will
not need in three lifetimes. So, I offer it up to you dear list.
Please make any offers for any of it, trades for anything useful are
especially welcome.

Ion pumps:

Vairan 20l/s triode pump, with controller and cables

Varian model 911-5032 30l/s triode pump with model 921-0062 controller
and cables

Varian model 924-0048 pressure relay box, no cables.

Diffusion pumps:

400 l/s 5-inch inside diameter diffusion pump with heater, cleaned and
dried, make unknown.

Tokyo Vacuum Machinery Co, Ltd. model OFK-3US 400 l/s with heater,
cleaned and dried. 5.25 inch inside diameter.

JEOL DP-4E 5.25 inch inside diameter, pump rate unknown, believed to be 400 l/s.

Other:

Pfeiffer turbo pump power supply, model TCP-100, has the heater-ring
thing with it, but no pump.

Link Analytical E.D.X.S. Supply control unit model 1108-059. This is
a box with relays and pneumatic valves in it for shutter control of a
TEM, I believe.

Panasonic KX-P1123 printer, compatible with Link EDS systems

1 monitor for a Link eXL system, working, but without cable.

1 Seiko Seiki STP control unit, model SCU-300H, no cables or pump.

I also have two complete JSM-35s worth of parts that I have collected,
so if anyone wants/needs parts for a JSM-35 or any ISI/Topcon parts,
let me know.

This collection is the result of acquiring multiple broken instruments
to get one working one. I've got everything I need to maintain the
working one for well beyond a reasonable amount of time.

Let me know if anything interests you, or it will probably end up on Ebay.

--Justin.



--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar

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From: ojanoc-at-ent.ufl.edu
Date: Tue, 20 Apr 2010 20:12:59 -0500
Subject: [Microscopy] viaWWW: SEM Need feedback on table-top SEM PHENOM

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Email: ojanoc-at-ent.ufl.edu
Name: Carol Ojano-Dirain

Organization: University of Florida

Title-Subject: [Filtered] SEM Need feedback on table-top SEM PHENOM

Message: I am considering buying the PHENOM table-top SEM and I would
appreciate feedback regarding the performance and ease of maintenance
for this piece of equipment. My applications typically range ~6,000x
to 10,000x.

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From: donc-at-asmicro.com
Date: Tue, 20 Apr 2010 23:22:37 -0500
Subject: [Microscopy] Re: [a] panhandling for volcanic dust

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Dear Frank,
I obtained a sample of Mt. St. Helens volcanic ash from an Indiana glass
artist (Joe Rice of Elwood IN) who uses it in glass paperweights to create a
veil decoration inside clear glass. Whereas normal pigments provide color,
the ash provides light scattering. A remarkable characteristic of the ash
is its slippery feel. Can anyone guess why glass might be slippery and what
particle shape you would expect to see?
regards,
Don
=============================================
Don Chernoff, Ph.D., President
Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com
3250 N. Post Rd., Ste. 120 Voice: 317-895-5630
INDIANAPOLIS IN 46226 USA Toll free: 800-374-8557 (in USA & Canada)
web: http://www.asmicro.com Fax: 317-895-5652
[business activities: analytical services in AFM, AFM probes, consulting,
training,
calibration and test specimens, calibration and measurement software,
used NanoScope equipment.] 2009-2010 is our 20th year in business!
=============================================
----- Original Message -----
From: Frank_Karl-at-lincolnelectric.com
To: donc-at-asmicro.com
Sent: Friday, April 16, 2010 7:17 AM
Subject: [a] [Microscopy] panhandling for volcanic dust





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I was listening to National public Radio on my way to work and one of the
big stories is how the volcanic ash from Iceland is disrupting air travel
in parts of Europe.

Of course as a light microscopist I would really enjoy getting a sample of
the dust, but a trip to Iceland or England is a little out of my reach,
even for a volcanic ash sample. Then I thought of you. That's the
collective you.

If anyone finds themselves sweeping or dusting a surface and grumbling
about the darn dust, if it's not too much trouble, could you sweep a
little
of it into a glassine or suitable container and mail me a sample? It
doesn't have to be too big and a few skin cells wouldn't hurt.

I'm not sure I can promise to reimburse you for the postage, but I will
put
your name on the slide and sometime in the future somebody will be looking
through my estate's slide collection (ebay maybe) wondering who you are.
Think of it as a limited form of immortality.

Thanks!

Frank Karl
PC32090
Lincoln Electric Company
22801 Saint Clair Ave.
Cleveland, Ohio USA.
44117-119,


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From: tbargar-at-unmc.edu
Date: Wed, 21 Apr 2010 09:08:04 -0500
Subject: [Microscopy] Need advice on using LR White

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Dear listers,

I am having a problem with micro-bubbles in my LR White. Whether I
polymerize the blocks in the oven or in the microwave I'm getting them. LR
White's poymerization is exothermic, but would this cause the bubbles,
which in turn are leaving holes in my thin sections. All advice and help
appreciated, thanks.


Tom Bargar
University of Nebraska Medical Center
Core Electron Microscopy Research Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

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From: naomi_mccallum-at-health.qld.gov.au
Date: Wed, 21 Apr 2010 17:38:36 -0500
Subject: [Microscopy] Fwd: viaWWW: SEM Need feedback on table-top SEM PHENOM

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Tom

I apologise if you already know this but are you polymerising in
gelatin capsules with most/all of the air excluded, because LR white
can be very fussy?

If it's stored too long it can have problems but I don't recall
bubbles.

Cheers

Malcolm

Malcolm Haswell
Electron Microscope Unit
Faculty of Applied Sciences
University of Sunderland
SUNDERLAND
SR1 3SD
UK

email: malcolm.haswell-at-sunderland.ac.uk

----- Original Message -----
X-from: tbargar-at-unmc.edu

Dear Listers

I am also interested in the response to Carol's post. Also is there anyone who has compared the Phenom to the JEOL JCM-5000 Neoscope distributed by Nikon?

Thanks in anticipation
Naomi


Naomi McCallum
Supervising Scientist, EM Unit, Anatomical Pathology
Pathology Queensland
_________________________________________________
Clinical and Statewide Services Division| Queensland Health

Block 7 Level 2
RBWH Queensland 4029
Ph: 07 3636 8057
Mob:
Fax: 07 3636 8908
Email: naomi_mccallum-at-health.qld.gov.au
Web: http://www.health.qld.gov.au/qhcss/qhps


} } } {ojanoc-at-ent.ufl.edu} 21/04/2010 11:21 am } } }



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Email: ojanoc-at-ent.ufl.edu
Name: Carol Ojano-Dirain

Organization: University of Florida

Title-Subject: [Filtered] SEM Need feedback on table-top SEM PHENOM

Message: I am considering buying the PHENOM table-top SEM and I would
appreciate feedback regarding the performance and ease of maintenance
for this piece of equipment. My applications typically range ~6,000x
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From: cmshelton-at-srggi.com
Date: Wed, 21 Apr 2010 18:15:41 -0500
Subject: [Microscopy] viaWWW: Light Microscope needed for Animal Shelter

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Email: cmshelton-at-srggi.com
Name: Cathy

Organization: SRGGI

Title-Subject: [Filtered] Light Microscope needed for Animal Shelter

Message: I am looking for someone to donate a decent light microscope
for our local animal shelter. This is a NO KILL shelter, funded by
the county. Most animal control shelters in the South, due to the
large influx of unwanted animals, are forced to euthenize. The
Lauderdale County Animal Shelter in Ripley, TN operates on a very
tight budget. This facility has undergone a huge transformation since
the new Mayor, with a huge heart for these unfortunate, unwanted
animals, took office a few years ago. They need a microscope to
check blood samples for heartworm microfilaria.
Thank you,
Cathy

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From: douglas.taatjes-at-uvm.edu
Date: Wed, 21 Apr 2010 18:16:11 -0500
Subject: [Microscopy] viaWWW: Announcement of Practical Stereology course at University

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Email: douglas.taatjes-at-uvm.edu
Name: Douglas J. Taatjes

Organization: University of Vermont

Title-Subject: [Filtered] Announcement of Practical Stereology course
at University of Vermont

Message: The Microscopy Imaging Center at the University of Vermont
will be hosting the following 3-day course on Practical Stereology:

What: Hands-on course in Practical Stereology
When: July 21-23, 2010
Where: Health Science Research Facility, University of Vermont, Burlington, VT

This 3 day practical course is designed for advanced students,
Post-Doctoral fellows, technicians, and Principal Investigators
seeking to understand the principles of stereology as applied to
microscopy and other imaging modalities. The course will be limited
to 20 participants.

Faculty:
Jens Nyengaard and Johnnie Andersen, Aarhus University Hospital, Denmark
Matthias Ochs, Hannover Medical School, Germany
Douglas J. Taatjes, University of Vermont, USA

Please refer to our website for registration forms and further
information, or contact Douglas Taatjes.

http://www.med.uvm.edu/microscopyimaging/


Douglas J. Taatjes
Microscopy Imaging Center
University of Vermont
Burlington, VT 05405
802-656-0373
douglas.taatjes-at-uvm.edu
http://www.med.uvm.edu/microscopyimaging/


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From: eikonika-at-otenet.gr
Date: Thu, 22 Apr 2010 01:46:39 -0500
Subject: [Microscopy] calibration of X axis in a JSM-5600LV SEM

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

Dear All

Our JSM-5600LV SEM produces images with 12.5% compression of the X axis. We
bought this scope second hand from the USA and we think this compression may
be on purpose; it converts the image from 4 : 3 to 3 : 2, because iif you
expand 12.5% the X axis to correct the compression you get:4+12.5% : 3 =
4.5 : 3 = 3 : 2
Whatever the explanation is, we would like to correct this X compression. Is
it posible to do it ourselves and if so, can anybody give us instructions
how to do it?

Thanks in advance

yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

eikonika-at-otenet.gr
yorgosnikas-at-hotmail.com
Tel/fax +30 210 8957677
Mobile +30 6945 107477


==============================Original Headers==============================
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7, 22 -- From: "yorgos nikas" {eikonika-at-otenet.gr}
7, 22 -- To: {microscopy-at-microscopy.com}
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From: P.Davies-at-swansea.ac.uk
Date: Thu, 22 Apr 2010 05:21:47 -0500
Subject: [Microscopy] Re: calibration of X axis in a JSM-5600LV SEM

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

I wondered whether the distortion may also be linked to slow scan controls
and TV technology, as I've learnt that TV standards eg PAL and NTSC don't
have square pixels like a 'normal' PC.
We have similar but distinct distortion issues to correct with our Japanese
(NTSC?) and European (PAL) SEMs.
FEI gave us a program for their SEM, but for our older JEOLs and their
add-on digital capture we use Photoshop to correct the images:

Image Menu -- Image size
Select "Resample Image" option, but deselect the "Constrain proportions"
option and then you will be able to add 12.5% to the 'width' pixels
DIMENSION box (not document), independently of the height dimension.

The 'actions' palette gives you a way of making an automated macro to adjust
them in bulk by recording the steps you've established to do this.


Remember to reset 'constrain proportions' when you've finished to avoid
distorting any other images you may be working with!

Best wishes,
Peter

Peter Davies
Scientific Imaging & Analysis facility
School of Engineering,
Swansea University
UK


On 22/04/2010 07:55, "eikonika-at-otenet.gr" {eikonika-at-otenet.gr} wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear All
}
} Our JSM-5600LV SEM produces images with 12.5% compression of the X axis. We
} bought this scope second hand from the USA and we think this compression may
} be on purpose; it converts the image from 4 : 3 to 3 : 2, because iif you
} expand 12.5% the X axis to correct the compression you get:4+12.5% : 3 =
} 4.5 : 3 = 3 : 2
} Whatever the explanation is, we would like to correct this X compression. Is
} it posible to do it ourselves and if so, can anybody give us instructions
} how to do it?
}
} Thanks in advance
}
} yorgos
}
} Dr Yorgos Nikas
} Athens Innovative Microscopy
} Skra 36 Voula 16673 GREECE
}
} eikonika-at-otenet.gr
} yorgosnikas-at-hotmail.com
} Tel/fax +30 210 8957677
} Mobile +30 6945 107477
}
}
} ==============================Original Headers==============================
} 7, 22 -- From eikonika-at-otenet.gr Thu Apr 22 01:46:38 2010
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} [85.74.228.252])
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} 7, 22 -- From: "yorgos nikas" {eikonika-at-otenet.gr}
} 7, 22 -- To: {microscopy-at-microscopy.com}
} 7, 22 -- Subject: calibration of X axis in a JSM-5600LV SEM
} 7, 22 -- Date: Thu, 22 Apr 2010 09:46:35 +0300
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12, 26 -- From P.Davies-at-swansea.ac.uk Thu Apr 22 05:21:46 2010
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12, 26 -- User-Agent: Microsoft-Entourage/12.24.0.100205
12, 26 -- Date: Thu, 22 Apr 2010 11:21:41 +0100
12, 26 -- Subject: Re: [Microscopy] calibration of X axis in a JSM-5600LV SEM
12, 26 -- From: Peter Davies {P.Davies-at-swansea.ac.uk}
12, 26 -- To: {eikonika-at-otenet.gr} , {microscopy-at-microscopy.com}
12, 26 -- Message-ID: {C7F5E1C5.7F00%p.davies-at-swansea.ac.uk}
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From: dsoren-at-umich.edu
Date: Thu, 22 Apr 2010 07:18:30 -0500
Subject: [Microscopy] buffer for TEM-DAB

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

Hello all,

Does anyone know if DAB reaction product is compatible with
Sorensen's buffer?

The reason I ask is that we are going to do some TEM processing of
DAB-stained tissue. The DAB staining will take place at another
site. Then the stained tissue will be shipped to us for TEM
processing. All of the references I have found use cacodylate buffer
for the initial fixation and later TEM prep. However, we want to
avoid shipping a hazardous material, and so would like to switch to
Sorensen's buffer before shipping. Can anyone foresee a problem with
doing this?

Thanks,
Dotty

Dorothy Sorenson
Microscopy and Image-analysis Laboratory
Department of Cell and Developmental Biology
University Of Michigan Medical School
A807 BSRB
109 Zina Pitcher Place
Ann Arbor, MI 48109-2200
(734)763-1170
FAX (734)763-1166



==============================Original Headers==============================
7, 17 -- From dsoren-at-umich.edu Thu Apr 22 07:18:30 2010
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7, 17 -- To: microscopy-at-microscopy.com
7, 17 -- From: Dorothy Sorenson {dsoren-at-umich.edu}
7, 17 -- Subject: buffer for TEM-DAB
7, 17 -- Date: Thu, 22 Apr 2010 08:18:27 -0400
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From: PhillipsT-at-missouri.edu
Date: Thu, 22 Apr 2010 07:40:11 -0500
Subject: [Microscopy] buffer for TEM-DAB

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

The DAB reaction is sensitive to the buffer. There are several papers showing that the reaction is strong (strongest?) in Imidazole buffer. Google Imidazole and DAB and they will pop up.

Thomas E. Phillips, Ph.D.
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
Biological Sciences
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (voice)
573-882-0123 (fax)


-----Original Message-----
X-from: dsoren-at-umich.edu [mailto:dsoren-at-umich.edu]
Sent: Thursday, April 22, 2010 7:19 AM
To: Phillips, Thomas E.

Hello all,

Does anyone know if DAB reaction product is compatible with
Sorensen's buffer?

The reason I ask is that we are going to do some TEM processing of
DAB-stained tissue. The DAB staining will take place at another
site. Then the stained tissue will be shipped to us for TEM
processing. All of the references I have found use cacodylate buffer
for the initial fixation and later TEM prep. However, we want to
avoid shipping a hazardous material, and so would like to switch to
Sorensen's buffer before shipping. Can anyone foresee a problem with
doing this?

Thanks,
Dotty

Dorothy Sorenson
Microscopy and Image-analysis Laboratory
Department of Cell and Developmental Biology
University Of Michigan Medical School
A807 BSRB
109 Zina Pitcher Place
Ann Arbor, MI 48109-2200
(734)763-1170
FAX (734)763-1166



==============================Original Headers==============================
7, 17 -- From dsoren-at-umich.edu Thu Apr 22 07:18:30 2010
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7, 17 -- To: microscopy-at-microscopy.com
7, 17 -- From: Dorothy Sorenson {dsoren-at-umich.edu}
7, 17 -- Subject: buffer for TEM-DAB
7, 17 -- Date: Thu, 22 Apr 2010 08:18:27 -0400
7, 17 -- X-Mailer: Apple Mail (2.753.1)
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==============================Original Headers==============================
16, 37 -- From PhillipsT-at-missouri.edu Thu Apr 22 07:40:11 2010
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16, 37 -- {microscopy-at-microscopy.com}
16, 37 -- Date: Thu, 22 Apr 2010 07:40:10 -0500
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From: larry.ackerman-at-ucsf.edu
Date: Thu, 22 Apr 2010 12:44:12 -0500
Subject: [Microscopy] buffer for TEM-DAB

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

I have found that some DAB protocols are sensitive to fading and so
always do a quick rinse and then postfix with osmium. Then the tissues
can go into a phosphate or other buffer for storage or dehydrated with
ethanol stopping at 70% ethanol for storage. [my DAB-Nickel-GOD protocol
uses 0.1M PO4 buffer].

Larry

PhillipsT-at-missouri.edu wrote:
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} The DAB reaction is sensitive to the buffer. There are several papers showing that the reaction is strong (strongest?) in Imidazole buffer. Google Imidazole and DAB and they will pop up.
}
} Thomas E. Phillips, Ph.D.
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} Biological Sciences
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (voice)
} 573-882-0123 (fax)
}
}
} -----Original Message-----
} X-from: dsoren-at-umich.edu [mailto:dsoren-at-umich.edu]
} Sent: Thursday, April 22, 2010 7:19 AM
} To: Phillips, Thomas E.
} Subject: [Microscopy] buffer for TEM-DAB
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hello all,
}
} Does anyone know if DAB reaction product is compatible with
} Sorensen's buffer?
}
} The reason I ask is that we are going to do some TEM processing of
} DAB-stained tissue. The DAB staining will take place at another
} site. Then the stained tissue will be shipped to us for TEM
} processing. All of the references I have found use cacodylate buffer
} for the initial fixation and later TEM prep. However, we want to
} avoid shipping a hazardous material, and so would like to switch to
} Sorensen's buffer before shipping. Can anyone foresee a problem with
} doing this?
}
} Thanks,
} Dotty
}
} Dorothy Sorenson
} Microscopy and Image-analysis Laboratory
} Department of Cell and Developmental Biology
} University Of Michigan Medical School
} A807 BSRB
} 109 Zina Pitcher Place
} Ann Arbor, MI 48109-2200
} (734)763-1170
} FAX (734)763-1166
}
}
}
} ==============================Original Headers==============================
} 7, 17 -- From dsoren-at-umich.edu Thu Apr 22 07:18:30 2010
} 7, 17 -- Received: from hellskitchen.mr.itd.umich.edu (smtp.mail.umich.edu [141.211.14.82])
} 7, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id o3MCIUpi028693
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} 7, 17 -- By hellskitchen.mr.itd.umich.edu ID 4BD03E93.B4BA2.29830 ;
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} 7, 17 -- To: microscopy-at-microscopy.com
} 7, 17 -- From: Dorothy Sorenson {dsoren-at-umich.edu}
} 7, 17 -- Subject: buffer for TEM-DAB
} 7, 17 -- Date: Thu, 22 Apr 2010 08:18:27 -0400
} 7, 17 -- X-Mailer: Apple Mail (2.753.1)
} ==============================End of - Headers==============================
}
}
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--
Larry Ackerman, Specialist
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From: protrain-at-emcourses.com
Date: Thu, 22 Apr 2010 12:56:37 -0500
Subject: [Microscopy] additional Monitors

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

Hi Listers

It's not often that I ask for your help but I am sure there is someone out
there who may have an answer for me?

I am working with a client who has two of the Hot FEG Hitachi SEM and each
of them only has one monitor. As a single monitor reduces the image area
they would like to fit an additional monitor so that one may be used as an
image area the other the control area. They have a problem in that they
cannot see a second monitor socket on the back of the HP PCs provided with
the instruments.

Does a lister have a Hitachi SU 70 or SU 6600 to which has been added an
additional monitor, if so how did you have to add a video driver or is my
client missing the second video out socket?

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



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From: patrick.mccurdy-at-colostate.edu
Date: Thu, 22 Apr 2010 18:13:19 -0500
Subject: [Microscopy] viaWWW: Mu metal and shielding column from EMI

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Email: patrick.mccurdy-at-colostate.edu
Name: Patrick McCurdy

Organization: Colorado State University

Title-Subject: [Filtered] Mu metal and shielding column from EMI

Message: We recently were forced to move our JSM6500F SEM. We now
have problems both with AC and DC EMI. Has anyone tried to shield
their column from EMI using a sheet of mu metal around the column,
instead of using a field cancelation system? Our system is almost
unusable at 1 kV.

Pat McCurdy
Colorado State University

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From: Patrick.Hogan-at-nasa.gov
Date: Thu, 22 Apr 2010 18:14:16 -0500
Subject: [Microscopy] viaWWW: Virtual Microscope Data

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Organization: NASA Ames Research Center

Title-Subject: [Filtered] Virtual Microscope Data

Message: We are looking for data to generate an open and growing
repository for microscopy specimens that can be viewed using the
Virtual Microscope (VM), http://virtual.itg.uiuc.edu/. We intend to
build a means for easily processing microscopy imagery into VM
specimen 'packages.' We plan to clearly identify the institution
and/or individual source of that data to always remain with the VM
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From: fahayes-at-ucdavis.edu
Date: Thu, 22 Apr 2010 21:39:39 -0500
Subject: [Microscopy]

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

Can someone suggest a sample prep method for mounting and/or coating
50nm-150nm sized Zinc Oxide particles suspended in water for SEM

Thanks in advance

Fred Hayes
Erica McKenzie
UC Davis


==============================Original Headers==============================
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4, 25 -- From: "Fred Hayes" {fahayes-at-ucdavis.edu}
4, 25 -- To: {Microscopy-at-microscopy.com}
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From: SWalck-at-SouthBayTech.com
Date: Fri, 23 Apr 2010 00:52:34 -0500
Subject: [Microscopy] Re:

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

I've never done ZnO particles, but I've done small diamond particles. The
particles were a little bigger than yours. I took a nebulizer and sprayed
the particles over a hot substrate. It was a multi-layer coating on glass
that was sitting on a hot plate. The idea was to dilute the particles
sufficiently and evaporate the water rapidly before the water drops could
touch other droplets. It worked fairly well. I then ion milled the
surface at a low angle and the diamond particles acted as a mask to allow
me to look at the multi-layers in the SEM. The particles stuck to the
surface very well.

If your particles are sufficiently deagglomerated and the solution is
dilute, I'll bet that this technique will work for you.


-Scott

Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA 92673

US Toll Free: 1-800-728-2233
Tel: (949) 492-2600
Fax: (949) 492-1499

www.southbaytech.com
SWalck-at-SouthBayTech.com


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} Can someone suggest a sample prep method for mounting and/or coating
} 50nm-150nm sized Zinc Oxide particles suspended in water for SEM
}
} Thanks in advance
}
} Fred Hayes
} Erica McKenzie
} UC Davis
}
}
} ==============================Original
} Headers==============================
} 4, 25 -- From fahayes-at-ucdavis.edu Thu Apr 22 21:39:39 2010
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} 4, 25 -- From: "Fred Hayes" {fahayes-at-ucdavis.edu}
} 4, 25 -- To: {Microscopy-at-microscopy.com}
} 4, 25 -- Cc: "Erica McKenzie" {ermckenzie-at-ucdavis.edu}
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From: nizets2-at-yahoo.com
Date: Fri, 23 Apr 2010 02:27:35 -0500
Subject: [Microscopy] Re:

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

Resuspending in Ethanol or methanol will increase the evaporation rate further.
Alternatively, you can try and find a suitable filter.



----- Original Message ----
X-from: "SWalck-at-SouthBayTech.com" {SWalck-at-SouthBayTech.com}
To: nizets2-at-yahoo.com
Sent: Fri, April 23, 2010 7:57:18 AM

I've never done ZnO particles, but I've done small diamond particles.  The
particles were a little bigger than yours.  I took a nebulizer and sprayed
the particles over a hot substrate.  It was a multi-layer coating on glass
that was sitting on a hot plate.  The idea was to dilute the particles
sufficiently and evaporate the water rapidly before the water drops could
touch other droplets.  It worked fairly well.  I then ion milled the
surface at a low angle and the diamond particles acted as a mask to allow
me to look at the multi-layers in the SEM.  The particles stuck to the
surface very well.

If your particles are sufficiently deagglomerated and the solution is
dilute, I'll bet that this technique will work for you.


-Scott

Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA  92673

US Toll Free: 1-800-728-2233
Tel: (949) 492-2600
Fax: (949) 492-1499

www.southbaytech.com
SWalck-at-SouthBayTech.com


}
}
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} Can someone suggest a sample prep method for mounting and/or coating
} 50nm-150nm sized Zinc Oxide particles suspended in water for SEM
}
} Thanks in advance
}
} Fred Hayes
} Erica McKenzie
} UC Davis
}
}
} ==============================Original
} Headers==============================
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From: jacques.faerber-at-ipcms.u-strasbg.fr
Date: Fri, 23 Apr 2010 04:35:42 -0500
Subject: [Microscopy] Re : ZnO particules for SEM

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Hi

If you have access to a plasma cleaner, I would take a piece of polished
dopped silicon (scrapp from microelectronics) and get it hyrdophilic by
a run in the plasma cleaner. Then you put a dropplet of your solution
more or less diluated with ethanol, methanol or acetone. The silicon
will give you a nice flat conductive surface, and there should bee no
need of a metallisation, using low kV for observation (less then 3 keV).
It's propably useful to ultrasonic the solution a bit after dilution. If
there is less enought water in it, the solution eaporate fast and the
particules shouldn't aglomerate too much. You will find big clsuters and
in between isolated particules.

Hope it helps

J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr



fahayes-at-ucdavis.edu a écrit :
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} Can someone suggest a sample prep method for mounting and/or coating
} 50nm-150nm sized Zinc Oxide particles suspended in water for SEM
}
} Thanks in advance
}
} Fred Hayes
} Erica McKenzie
} UC Davis
}
}
} ==============================Original Headers==============================
} 4, 25 -- From fahayes-at-ucdavis.edu Thu Apr 22 21:39:39 2010
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From: wvrenter-at-sckcen.be
Date: Fri, 23 Apr 2010 07:11:23 -0500
Subject: [Microscopy] viaWWW: electropolisher

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Email: wvrenter-at-sckcen.be
Name: Wouter Van Renterghem

Organization: SCKïCEN

Title-Subject: [Filtered] electropolisher

Message: I would like to purchase a new
electropolisher for TEM applications. I am
chosing between a Struers Tenupol-5 and a
Fischione model 110. Can anyone comment on the
performance these instruments?

Thank you,
Wouter Van Renterghem

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From: dstumpf-at-hersheys.com
Date: Fri, 23 Apr 2010 07:11:59 -0500
Subject: [Microscopy] viaWWW: Re: Mu metal and shielding column from EMI

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Email: dstumpf-at-hersheys.com
Name: Dave Stumpf

Organization: The Hershey Company

Title-Subject: [Filtered] Re: Mu metal and shielding column from EMI

Message: I had a mu metal shield on my JSM 6300 that was provided and
installed by JEOL. The installation engineer said it made a
difference. That scope is gone but I still have the shield.

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From: Woody.White-at-areva.com
Date: Fri, 23 Apr 2010 07:19:30 -0500
Subject: [Microscopy] viaWWW: Mu metal and shielding column from EMI

Contents Retrieved from Microscopy Listserver Archives
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Pat,

I have not used mu metal around the column, but was successful in
reducing interference levels, in an past situation, by (mostly)
enclosing the chamber working distance with a mu metal cylinder. Of
course, had to leave some room for signal electron exit.

Along this line, if the interference is significant, shielding only the
column may not be enough. The chamber (or in my case the W.D. electron
path) may need to also be shielded.

FYI: For the most effective mu metal shield, it must not be work
hardened. That is, if bent, it should be re-annealed.

Woody


N.W. (Woody) White Jr.
Senior Electron Microscopist
Chemistry and Materials Center
AREVA NP Inc
An AREVA and Siemens Company
Lynchburg, VA
Lab Phone: 434.832.3004

-----Original Message-----
X-from: patrick.mccurdy-at-colostate.edu
[mailto:patrick.mccurdy-at-colostate.edu]
Sent: Thursday, April 22, 2010 7:25 PM
To: WHITE Norvell (AREVA NP INC)

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Email: patrick.mccurdy-at-colostate.edu
Name: Patrick McCurdy

Organization: Colorado State University

Title-Subject: [Filtered] Mu metal and shielding column from EMI

Message: We recently were forced to move our JSM6500F SEM. We now have
problems both with AC and DC EMI. Has anyone tried to shield their
column from EMI using a sheet of mu metal around the column, instead of
using a field cancelation system? Our system is almost unusable at 1 kV.

Pat McCurdy
Colorado State University

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From: DusevichV-at-umkc.edu
Date: Fri, 23 Apr 2010 09:43:58 -0500
Subject: [Microscopy] RE:

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

I do not know about Zn oxide, but usually for particles I start with the
simplest method: put a droplet of suspension on a glass, or even better
on a plastic coverslip. Plastic is better for BSE visualization. You can
also use TEM grids with carbon film and mount them on carbon substrate
if you want better EDS. Some experimentation with concentration of
suspension may be needed. Coating depends on your equipment: if you
microscope capable of high magnifications at low voltages, do not coat.
Otherwise coat as usual. If you are planning BSE and/or EDS, try carbon
coating.

Good luck,

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

} -----Original Message-----
} From: fahayes-at-ucdavis.edu [mailto:fahayes-at-ucdavis.edu]
} Sent: Thursday, April 22, 2010 9:41 PM
} To: Dusevich, Vladimir
} Subject: [Microscopy]
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}
} Can someone suggest a sample prep method for mounting and/or coating
} 50nm-150nm sized Zinc Oxide particles suspended in water for SEM
}
} Thanks in advance
}
} Fred Hayes
} Erica McKenzie
} UC Davis
}
}
} ==============================Original
} Headers==============================
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From: jteshima-at-dunesciences.com
Date: Fri, 23 Apr 2010 10:18:42 -0500
Subject: [Microscopy] Re:

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


Have a look at Dune Sciences video showing how to prepare nanoparticle
suspended in solution directly on to SMART Grids. The grids have been
functionalized to have a hydrophilic surface that can also be
positively or negatively charged. In the case of gold nanopartilcles
in a citrate solution positively charged Nanoplus grids were used.
The images shown are TEM but we have also imaged particles in the SEM,
this is an especially nice method if you also have a STEM detector in
your microscope. You can use the silicon surface for SE imaging and
get better resolution imaging in STEM mode through the thin part of
the membrane.
http://www.dunesciences.com/prep_video.html

There is also a white paper written by Kurt Langworthy of the
University of Oregon, "Nanoparticle Analysis" that also describes this
technique. http://www.dunesciences.com/

Janet Teshima
Dune Sciences
503-544-7526
jteshima-at-dunesciences.com

On Apr 23, 2010, at 12:37 AM, nizets2-at-yahoo.com wrote:

}
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} Resuspending in Ethanol or methanol will increase the evaporation
} rate further.
} Alternatively, you can try and find a suitable filter.
}
}
}
} ----- Original Message ----
} X-from: "SWalck-at-SouthBayTech.com" {SWalck-at-SouthBayTech.com}
} To: nizets2-at-yahoo.com
} Sent: Fri, April 23, 2010 7:57:18 AM
} Subject: [Microscopy] Re:
}
}
}
}
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} America
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}
} I've never done ZnO particles, but I've done small diamond
} particles. The
} particles were a little bigger than yours. I took a nebulizer and
} sprayed
} the particles over a hot substrate. It was a multi-layer coating on
} glass
} that was sitting on a hot plate. The idea was to dilute the particles
} sufficiently and evaporate the water rapidly before the water drops
} could
} touch other droplets. It worked fairly well. I then ion milled the
} surface at a low angle and the diamond particles acted as a mask to
} allow
} me to look at the multi-layers in the SEM. The particles stuck to the
} surface very well.
}
} If your particles are sufficiently deagglomerated and the solution is
} dilute, I'll bet that this technique will work for you.
}
}
} -Scott
}
} Scott D. Walck, Ph.D.
} Technical Director
} South Bay Technology, Inc.
} 1120 Via Callejon
} San Clemente, CA 92673
}
} US Toll Free: 1-800-728-2233
} Tel: (949) 492-2600
} Fax: (949) 492-1499
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} www.southbaytech.com
} SWalck-at-SouthBayTech.com
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} }
} }
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} }
} } Can someone suggest a sample prep method for mounting and/or coating
} } 50nm-150nm sized Zinc Oxide particles suspended in water for SEM
} }
} } Thanks in advance
} }
} } Fred Hayes
} } Erica McKenzie
} } UC Davis
} }
} }
} } ==============================Original
} } Headers==============================
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Janet Teshima
SMART Grids Product Manager
cell: 503-5447526
office: 541-636-3712
jteshima-at-dunesciences.com




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From: maryflet-at-interchange.ubc.ca
Date: Fri, 23 Apr 2010 11:00:12 -0500
Subject: [Microscopy] viaWWW: electropolisher

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Dear Wouter,
We have run a Struers Tenopol-2 since 1984 and it is still working. It seems
to be very robust. We have replaced the sample holders and repaired it a few
times, but it has lasted very well, considering its constant contact with
corrosive electropolishing solutions and use by students. I have no
experience with the Fischione electropolisher, but I have purchased other
materials from Fischione and they have worked just fine.

Regards,

Mary Fletcher
Electron Microscopist
Materials Eng. UBC
#309 - 6350 Stores Road
Vancouver, B.C. V6T 1Z4
Canada
Tel: 604-822-5648
Fax: 604-822-3619
email: maryflet-at-interchange.ubc.ca


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Email: wvrenter-at-sckcen.be
Name: Wouter Van Renterghem

Organization: SCKïCEN

Title-Subject: [Filtered] electropolisher

Message: I would like to purchase a new
electropolisher for TEM applications. I am
chosing between a Struers Tenupol-5 and a
Fischione model 110. Can anyone comment on the
performance these instruments?

Thank you,
Wouter Van Renterghem

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From: kenconverse-at-qualityimages.biz
Date: Fri, 23 Apr 2010 11:32:48 -0500
Subject: [Microscopy] viaWWW: Mu metal and shielding column from EMI

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Pat,
You say it's almost unusable at 1kV. At what kind of mag? Perhaps you
should try and go after the source of the EMI and see if you can reduce it.
Mu metal around the column (and chamber) can be helpful, but if the problem
is really severe, it may not do what you want. If the source is localized,
it may be easier to shield than the column, although the shielding JEOL
provides works pretty well.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


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Email: patrick.mccurdy-at-colostate.edu
Name: Patrick McCurdy

Organization: Colorado State University

Title-Subject: [Filtered] Mu metal and shielding column from EMI

Message: We recently were forced to move our JSM6500F SEM. We now
have problems both with AC and DC EMI. Has anyone tried to shield
their column from EMI using a sheet of mu metal around the column,
instead of using a field cancelation system? Our system is almost
unusable at 1 kV.

Pat McCurdy
Colorado State University

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From: kjl226-at-vt.edu
Date: Fri, 23 Apr 2010 15:09:17 -0500
Subject: [Microscopy] viaWWW: McDowells fixative

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Email: kjl226-at-vt.edu
Name: Kathy J. Lowe

Organization: Vet. School at Virginia Tech

Title-Subject: [Filtered] McDowells fixative

Message: Does anyone know how to make McDowells fixative?

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From: lamiller-at-illinois.com
Date: Fri, 23 Apr 2010 15:09:52 -0500
Subject: [Microscopy] viaWWW: SEM Prep - particles

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Email: lamiller-at-illinois.com
Name: Lou Ann Miller

Organization: U of Illinois

Title-Subject: [Filtered] SEM Prep - particles

Message: I've done cells and particles before



I put a syringe filter (13 mm) on a 1-3cc syringe, And gently
filter a diluted solution of particles/ cells etc through.


Then I use the same syringe filter setup to put the processing
chemicals through, if I use HMDS, I remove the syringe and
process the filter unit in a scintillation vial, seems to work fine,
though times are not excessive.

Then dry, even in mild vacuum, and either disemble or cut out the
filter to mount and coat.


At some points the filter may crack , or I intentionally use a
hyperdermic needle to create one small hole up the exit port into
the filter, to assist when gently pushing fluids through becomes
hard. This still works fine.

The final product is on the filter, in pretty good quantity even with
scarce samples, though it will have the hills and valleys of the
filter ridges. This could be good or bad depending, sometimes that
even helps.

For coating I stand the stubs at different angles for multiple short coatings.


Hope that Helps,


Lou Ann


{ { { { { { { { {} } } } } } } } } } } } } }
Lou Ann Miller, Service Supervisor
Center for Microscopic Imaging
College of Veterinary Medicine
University of Illinois MC=002

Room 1204 VMBSBld
2001 S Lincoln Ave
Urbana, IL 61821

217-244-1567
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From: W.Muss-at-salk.at
Date: Sat, 24 Apr 2010 04:02:03 -0500
Subject: [Microscopy] Re: McDowells fixative

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Good morning from Europe,

dear Kathy,
perhaps it seems to me you don't have the opportunity to google anyway, since there are a lot of positive sources when searching for " mcdowell AND Fixative " (approx. 48,000 results).
As a Member of MSA in good standing I would like to help just for the case there are no replies on the listserver (as of Sat, 24th of Apr,2010, 10:25 a.m. EST (Greenwich + 1 h) (not knowing wether this is an IT-problem on my side or a problem of the List server system, since I have noticed to have not got some original questions but only some, but not many replies on several subjects within the last days)....


McDOWELL/TRUMP-4(C*))F:1G FIXATIVE is a commonly used fixative for EM (or at least specimen preparation for resin embedded semithin sections)
F stands for "formaldehyde". "G" for glutaraldehyde.
"4" = 4% formaldehyde "1" = 1% glutaraldehyde
[*) "4CF" stands for: 4% "commercial" formaldehyde]

but it seems that is tricky to get rapidly correct recipes for this

on: http://www.biotech.ufl.edu/EM/data/trump.html e.g. you can find:
---------------------------------------------------------------------------
Trumps Fixative **)
Ten ml of 40% formaldehyde and 2 ml of 50% glutaraldehyde were added to 200-mOsm buffer.

This solution could be conveniently prepared for routine use as follows:

1.16 g of NaH[2]PO[4] is added to 88 ml water and dissolved.

Add 0.27 g NaOH followed by the formaldehyde and the glutaraldehyde.
----------------------------------------------------------------------------
**) you perhaps cannot follow this recipe because ist formulation/instruction is not quite complete:
i) ...were added to (a) 200 mOsm buffer to result in 4% formaldehyde and 1 % glutaraldehyde, which is
ii) for 100 ml fixative: solve 1.16 g NaH[2]PO[4] in -say- 80 ml A.dest, take care for a complete dissolution of the powder; if this is accomplished, add 0.27 g NaOH (means sodium hydroxide 'pastilles', which is not quite easy to weigh out) and let solve completely by stirring, then add 10-12 ml of 38% formaldehyde solution (normally you won't have really a 40% FA-solution; if there is a pH - acid stabilizing powder in the bottle it is recommended to filter; don't use old batches of FA as well as GA), stir, then add 2 ml of 50% GA (or 4 ml of 25% GA), mix and then fill up the volume with A.dest up to 100 ml.

This fixative named McDowell/Trump-Fixative has been praised in the late 70ies and until the mid 80ies of last century for routine fixation when performing (correlative) Light-histological paraffin embedded - resin semithin-LM and (T)EM-diagnostic specimens and the original article as well as other sources/references recommend a making of solution in volumes up to some gallons in stock...(depending on the daily demand). Unfortunately it seems that this mixture did not make it to the top...the main fixative in diagnostic light histological specimen preparation and diagnosis still is (at least buffered) 4-6 % formaldehyde solution.

The original article is:

McDowell, E and B. Trump. 1976. Histological fixatives for diagnostic light and electron microscopy. Arch. Pathol. Lab. Med. 100:405-414
If you need the original paper I could provide this within a pdf.

Note added:
=======================================
from MSA-Listserver Archives 2004:

FIXATION -Trump's fixative
I work for a small research lab and we plan to do TEM on mouse tissue. We will fix the tissue in Trumps which is 4% formaldehyde + 1 % glutaraldehyde in a phosphate buffer. Has anyone used this fixative before? Are there any pitfalls that we should avoid?
Scott Duong {sduong-at-bidmc.harvard.edu} 16 Aug 2004
Re:
Phosphate fixes can result in fine calcium precipitates. I prefer HEPES orPIPES.
Tom Phillips {phillipst-at-missouri.edu} 16 Aug 2004

Re:
I use a modified Trump's fixative with 0.1 M sodium cacodylate buffer without problems with most kinds of tissue. I also use 0.1 M phosphate butter without artifact problems, as lang as I wash in buffer 3X after fixation prior to osmification (1 % osmium in 0.1 M buffer for 60 minutes), followed by 3X rinse in buffer and an additional 3X rinse in waterprior to dehydration in ethanol series. My problem had been "osmium peppering" -- a fine precipitate contaminating samples run up in phosphate buffer when not thoroughly washed. Sodium cacodylate avoids this problem, but I always rinse thoroughly when preparing samples tor TEM. I run up tissue at room temperature because of a superstition acquired as a graduate student that cold temperature promotes the disassembly of microtubules. Does anyone have thoughts pro or con about running up tissue on ice vs room temperature?
Dean Abel {dean-abel-at-uiowa.edu} 17 Aug 2004

Re:
I have used a combination fixative of 1 % glutaraldehyde + 1 % Os04 in 0.05 M phosphate buffer at pH 6.2 on ice and kept in the dark for 45 minutes for many tissues. There are many microtubules present.
Perhaps the microtubules were stabilized by the osmium before they were able to disassemble in the cold temperature. The only problem I had with this fixative was with bacteria inside tissue cultured macrophages. It did not wash out as easily/quickly as I expected. As a result, I had the "peppering" of the osmium in and around the bacteria itself when the TEM beam hit the bacteria.
Pat Connelly {psconnel-at-sas.upenn.edu} 17 Aug 2004
============================================

Best wishes, good luck and
sincerely yours,


Wolfgang MUSS
SALZBURG
EM-Lab, Pathology
SALK-LKH/PMU


} -----Ursprüngliche Nachricht-----
} Von: kjl226-at-vt.edu [mailto:kjl226-at-vt.edu]
} Gesendet: Freitag, 23. April 2010 22:14
} An: Muß Wolfgang
} Betreff: [Microscopy] McDowells fixative
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} Does anyone know how to make McDowells fixative?
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From: singinggardenersx2-at-live.com
Date: Sat, 24 Apr 2010 08:00:30 -0500
Subject: [Microscopy] viaWWW: McDowells fixative

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Email: singinggardenersx2-at-live.com
Name: Gigi Kemalyan

Organization: San Joaquin Delta College

Title-Subject: [Microscopy] viaWWW: McDowells fixative‏

Message: Hi Kathy,

If you are referring to McDowell & Trump 4F:1G, an informative
article including the recipe can be found on page 50 in the March
2010 issue of Microscopy Today.

http://content.yudu.com/A1n109/MTOVol18Issue2/resources/index.htm?referrerUrl=http%3A%2F%2Fwww.microscopy-today.com%2F

Sincerely,

Gigi Kemalyan
EM Student, San Joaquin Delta College
Stockton, California

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From: s.h.coetzee-at-gmail.com
Date: Sat, 24 Apr 2010 13:25:54 -0500
Subject: [Microscopy] Re: Re : ZnO particules for SEM

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

As been mentioned, you need a conductive surface and the Si route
should work fine.
There are other quick methods that do work if you have a reasonable
amount of particles to play with.

I have used Formavar coated TEM grids in the past with reasonable
amount of success (Au, and Ag aprticles and clays from suspention)
particles and a lot quicker of you have Holy carbon grids (not lacy)
in your EM unit. I also did some work on plain polished SEM stubs.
One of my favourites is SPI sticky tabs. (from my experience the other
manufacturers are not a sticky). We left some of them for two weeks
in the field to collect dust particles. Just glue down on a stub,
drop the solution on, let it evaporate and if you are fortunate, happy
viewing. We are currently also investigating different filers and we
hope to get to a good suggestion soon.

Have fun (that is what microscopy is all about!)

Stephan Coetzee


On Fri, Apr 23, 2010 at 11:44 AM, {jacques.faerber-at-ipcms.u-strasbg.fr} wrote:
}
}
}
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} Hi
}
} If you have access to a plasma cleaner, I would take a piece of polished
} dopped silicon (scrapp from microelectronics) and get it hyrdophilic by
} a run in the plasma cleaner. Then you put a dropplet of your solution
} more or less diluated with ethanol, methanol or acetone. The silicon
} will give you a nice flat conductive surface, and there should bee no
} need of a metallisation, using low kV for observation (less then 3 keV).
} It's propably useful to ultrasonic the solution a bit after dilution. If
} there is less enought water in it, the solution eaporate fast and the
} particules shouldn't aglomerate too much. You will find big clsuters and
} in between isolated particules.
}
} Hope it helps
}
} J. Faerber
} IPCMS-GSI
} (Institut de Physique et Chimie des Matériaux de Strasbourg
} Groupe Surface et Interfaces)
} 23, rue de Loess ; BP43
} 67034 Strasbourg CEDEX 2
} France
}
} Tel 00 33(0)3 88 10 71 01
} Fax 00 33(0)3 88 10 72 48
} E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr
}
}
}
} fahayes-at-ucdavis.edu a écrit :
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} }
} } Can someone suggest a sample prep method for mounting and/or coating
} } 50nm-150nm sized Zinc Oxide particles suspended in water for SEM
} }
} } Thanks in advance
} }
} } Fred Hayes
} } Erica McKenzie
} } UC Davis
} }
} }
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}



--
Stephan H Coetzee
Chief Technician
Electron Microscope Unit
University of Botswana


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From: protrain-at-emcourses.com
Date: Sat, 24 Apr 2010 13:35:02 -0500
Subject: [Microscopy]

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

Hi

The simple method that I would use is to coat a cover slip with gold and
then to deposit "blobs" of the media having given it a chance to mix through
placing in an ultrasonic bath for a few minutes.

As the glass should be perfectly clean all that you will see are your
particles. With very fine structures and low kV there is often no need to
coat with this method.


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com


-----Original Message-----
X-from: fahayes-at-ucdavis.edu [mailto:fahayes-at-ucdavis.edu]
Sent: 23 April 2010 03:41
To: protrain-at-emcourses.com

Can someone suggest a sample prep method for mounting and/or coating
50nm-150nm sized Zinc Oxide particles suspended in water for SEM

Thanks in advance

Fred Hayes
Erica McKenzie
UC Davis


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From: protrain-at-emcourses.com
Date: Sat, 24 Apr 2010 13:35:07 -0500
Subject: [Microscopy] Re: Additional Monitors

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

Hi All

I would like to thank all those who took the trouble to answer my email,
some of whom clearly passed my question on the their IT department!

A special thanks to the Hitachi boys, on the ball as usual, Bill Roth and
Mark Betts.

But to you all from my client and I, many thanks.

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



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From: kraftpiano-at-gmail.com
Date: Sun, 25 Apr 2010 00:49:41 -0500
Subject: [Microscopy] Moving an SEM.

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

About a year ago, I started talking with a company out in California
who wanted to donate their SEM to a school, but couldn't find someone
willing to help them link it up to the school.  It's a JSM-35C, and
since I've already got so many parts and a lot of experience with
those, I took on the task.

A year later, it was ready to be sent to me.  It was due to arrive at
about the same time the JSM-6100 from Toledo arrived, so I would have
two of them in the same room working on them at the same time.  Not a
problem, I spent two weeks cleaning out the garage and getting it
sealed and ready for the shipments.

The shipping of the 6100 from Toledo was the most pleasant experience
I have ever had moving large equipment.  The guys in Ohio did an
excellent job packing it up and readying it for shipment.

The JSM-35C, however, is still not here.  The shipping company is
making every effort to locate it, but so far has had no luck.  It's
been at least three weeks now.

My question is: Surely I can't be the only one who has had a moving
horror story with an EM, but I can't really imagine how it is possible
to lose 2500 pounds of analytical equipment.

If anyone else has some horror stories, I'd love to hear them.

Thanks,

Justin.

--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar


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From: ivan.baines-at-maxplanckflorida.org
Date: Mon, 26 Apr 2010 07:53:54 -0500
Subject: [Microscopy] viaWWW: Job opportunity for Electron Microscopist

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

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
---------------------------------------------------------------------------
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Email: ivan.baines-at-maxplanckflorida.org
Name: Ivan C. Baines

Organization: Max Planck Florida Institute

Title-Subject: [Filtered] Job opportunity for Electron Microscopist

Message: The Max Planck Florida Institute (MPFI) is a new institute
of the renowned Max Planck Society of Germany and the first Max
Planck Institute in the USA. This is an opportunity for an inspired
and motivated electron microscopist with at least 5 years experience
covering a range of EM techniques to design, setup and run a
state-of-the-art EM Facility. The MPFI intends to be a powerhouse of
imaging and will invest in 3 electron microscopes together with the
appropriate supporting instrumentation to permit TEM, tomography,
HPF, AFS, frozen thin section immuno-EM and the exploration of new
techniques such as serial block facing SEM. More info to be found at
the MPFI www site job listings:

http://www.maxplanckflorida.org/employment.html?jobid=63

Please feel welcome to contact me for further details.

Ivan Baines
Chief Scientific Facilities Officer

Login Host: 99.99.2.115
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From: oshel1pe-at-cmich.edu
Date: Mon, 26 Apr 2010 11:32:00 -0500
Subject: [Microscopy] Fwd: Re: Ask-A-Microscopist

Contents Retrieved from Microscopy Listserver Archives
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==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
(Not to me - Oshel)
==========================================================
I have already suggested ImageJ, but Safari wishes to contact someone
who does particle tracking.
Phil

Thank you so much for answering me. I want to work with "Confocal
microscope"(light microscopy), and the main target of my project is
to use a software to analyze the results taken from microscope for
"nanoparticle tracking" which helps me to calculate some biophysical
properties.

Looking forward to hearing from you,
Regards,
Fatemeh

realname - Fatemeh Safari

Email -
{mailto: {mailto: {mailto:safari.fatemeh-at-gmail.com} safari.fatemeh-at-gmail.com} {mailto:safari.fatemeh-at-gmail.com} safari.fatemeh-at-gmail.com} {mailto: {mailto:safari.fatemeh-at-gmail.com} safari.fatemeh-at-gmail.com} {mailto:safari.fatemeh-at-gmail.com} safari.fatemeh-at-gmail.com


ORGANIZATION - NUS
EDUCATION - Graduate College
LOCATION - Singapore
SUBJECT_OF_QUESTION - Nanoparticle tracking software
QUESTION - Dear Mr/Mrs

I'm novice researcher in nanoparticle imaging and tracking fields.
I'm searching for comercial software which provides the best features
among all existing ones.
could you please kindly help me.

Thank you


--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576
Microscopy Society of America
Ask a Microscopist
http://www.microscopy.org/microscopy/ask.cfm

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From: ian.drucker-at-gmail.com
Date: Tue, 27 Apr 2010 08:26:28 -0500
Subject: [Microscopy] Measurements

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

I'm looking for a free ready to use program that can do measurements
of Tif images from a SEM. I know there's several free image
manipulation packages out there but I haven't found a ready to use
measurement function that can take into account the image
magnification and such.

Anyone with some suggestions?


Thanks

==============================Original Headers==============================
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From: James.Passmore-at-sealedair.com
Date: Tue, 27 Apr 2010 10:00:10 -0500
Subject: [Microscopy] Re: Measurements

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

ian.drucker-at-gmail.com wrote on 04/27/2010 09:28:49 AM:
}
} I'm looking for a free ready to use program that can do measurements
} of Tif images from a SEM. I know there's several free image
} manipulation packages out there but I haven't found a ready to use
} measurement function that can take into account the image
} magnification and such.
}
} Anyone with some suggestions?
}
}
} Thanks
}

ImageJ
Free, extensible, and has a great user community.
http://rsbweb.nih.gov/ij/


----------------------------------------------
Jim Passmore
Research Associate
Sealed Air Corporation
james.passmore-at-sealedair.com
----------------------------------------------



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From: ehaller-at-health.usf.edu
Date: Tue, 27 Apr 2010 10:23:57 -0500
Subject: [Microscopy] Measurements

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

Hi, Ian,

I can't offer much help, I'm afraid. I'm in the same boat. I'm writing to ask that if you receive replies off the listserver that you please post them to the listserver. I have a project of 60 SEM samples of sharkskin where we need to measure dermal scale distance on tiff images. I'm aware of the arguments against accuracy of measurements from SEM images, but we're doing low mag work on basically flat images at essentially zero tilt. We don't have the photos yet. I have a couple of old calibration standards from Ladd and Fullam to calibrate the mag in my SEM. I've checked it before and the readout was accurate.
I was going to fool around with Photoshop, a program called FijiImage, which is an offshoot of ImageJ, and then try to import the tiffs into my TEM where I have Olympus Soft Imaging software (AnalySis), but that software is calibrated for my TEM mag's so, I would have a lot of work there to recalibrate it for the SEM mags before I could use its measure function. Those are the options I have open.

FijiImage can be downloaded at: http://pacific.mpi-cbg.de/wiki/index.php/Main_Page

For those ImageJ users, I've forgotten how to load and open plugins. I know there's a measure plug-in that can be downloaded and a way to calibrate ImageJ for microns. Maybe you can tell us where it is and walk Ian and I through loading and using it in a non-technical way. I'm user-manual challenged :)

Ed

Edward Haller, Lab Manager
Integrative Biology Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-cas.usf.edu

________________________________________
X-from: ian.drucker-at-gmail.com [ian.drucker-at-gmail.com]
Sent: Tuesday, April 27, 2010 9:37 AM
To: Haller, Edward

I'm looking for a free ready to use program that can do measurements
of Tif images from a SEM. I know there's several free image
manipulation packages out there but I haven't found a ready to use
measurement function that can take into account the image
magnification and such.

Anyone with some suggestions?


Thanks

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From: rcsencsits-at-lbl.gov
Date: Tue, 27 Apr 2010 10:41:02 -0500
Subject: [Microscopy] Re: Moving an SEM.

Contents Retrieved from Microscopy Listserver Archives
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Hi Justin,

Two years ago a colleague and I went to work with JEOL engineer to
dismantle and pack up to ship a JEOL 4000. We crated everything and
watched the moving company load their truck, everything arrived a week
later. Sorry no horror. We reattached most things, had JEOL take
care of the high voltage tank and when we turned it on--beam!! As it
should be.

More details upon request.
Best regards,
Roseann


Roseann Csencsits, PhD
Scientist in Charge - Donner TEM Facility
Lawrence Berkeley Lab 01-365
1 Cyclotron Road
Berkeley, CA 94720
510-486-4548







On Apr 24, 2010, at 11:08 PM, kraftpiano-at-gmail.com wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} About a year ago, I started talking with a company out in California
} who wanted to donate their SEM to a school, but couldn't find someone
} willing to help them link it up to the school. It's a JSM-35C, and
} since I've already got so many parts and a lot of experience with
} those, I took on the task.
}
} A year later, it was ready to be sent to me. It was due to arrive at
} about the same time the JSM-6100 from Toledo arrived, so I would have
} two of them in the same room working on them at the same time. Not a
} problem, I spent two weeks cleaning out the garage and getting it
} sealed and ready for the shipments.
}
} The shipping of the 6100 from Toledo was the most pleasant experience
} I have ever had moving large equipment. The guys in Ohio did an
} excellent job packing it up and readying it for shipment.
}
} The JSM-35C, however, is still not here. The shipping company is
} making every effort to locate it, but so far has had no luck. It's
} been at least three weeks now.
}
} My question is: Surely I can't be the only one who has had a moving
} horror story with an EM, but I can't really imagine how it is possible
} to lose 2500 pounds of analytical equipment.
}
} If anyone else has some horror stories, I'd love to hear them.
}
} Thanks,
}
} Justin.
}
} --
} "America believes in education; the average professor earns more money
} in a year than a professional athlete earns in a whole week." Evan
} Esar


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From: frah0010-at-umn.edu
Date: Tue, 27 Apr 2010 12:05:41 -0500
Subject: [Microscopy] Ba in X-glass

Contents Retrieved from Microscopy Listserver Archives
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Microscopy colleagues,

Does anyone know of a measured value for the Ba concentration in the X-Glass analytical standard from Corning?

Thanks,
Ellery

-----------------
Ellery Frahm
Senior Research Fellow, Department of Geology & Geophysics
Manager & Principal Analyst, Electron Microprobe Lab
Doctoral Candidate, Department of Anthropology
University of Minnesota - Twin Cities campus
Lab website: http://probelab.geo.umn.edu/
Personal: http://web.mac.com/elleryfrahm/

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From: ressci1-at-gmail.com
Date: Tue, 27 Apr 2010 14:19:33 -0500
Subject: [Microscopy] Olympus IX70/CARV spinning disk confocal needs new home

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Hi all,
Due to a lab re-sizing, we currently have an Olympus IX70 microscope
equipped with a CARV spinning disk confocal system that needs a new home.
Comes with computer and Metamorph software. We are looking for offers.
Please contact Erin off-list for photos, configuration, and additional
details. System is in San Diego but we can ship anywhere.
Thanks,
Erin
ressci1-at-gmail.com
510-344-6633

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From: hyi-at-emory.edu
Date: Wed, 28 Apr 2010 06:36:18 -0500
Subject: [Microscopy] GLP EM Lab

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Dear All:

We have been approached to take on a major client/project that requires
us to be GLP (Good Laboratory Practice) compliant. Does anyone know a
university EM facility that is a GLP lab. We would like to get some advice.
Thank you in advance.

Hong
Emory Robert P. Apkarian IEMC
(404) 712-8491


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From: mcintyre-at-optics.rochester.edu
Date: Wed, 28 Apr 2010 13:43:17 -0500
Subject: [Microscopy] Student Project Website

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Hi Listers-

We've nearly concluded the semester at Rochester, the trees are
blooming, the birds are singing, its not below freezing at night
anymore...so it must be time for Electron Microscopy Practicum student
projects.

If you care to take a look and comment on their successes or their
"opportunities to excel" please visit here:

http://www.optics.rochester.edu/workgroups/cml/opt307/spr10/

Thanks!
Brian
--

Brian McIntyre
Lab Manager URinc
Univ. of Rochester
Institute of Optics
Wilmot Bldg.
Rochester, NY 14627

585-275-3058 (office)
585-244-4936 (fax)
585-301-3145 (cell)

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From: rbeavers-at-mail.smu.edu
Date: Wed, 28 Apr 2010 13:46:06 -0500
Subject: [Microscopy] Dallas area access to a surface area mesurement tool BET

Contents Retrieved from Microscopy Listserver Archives
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Group,

Have a student searching for a BET tool.

If you have any leads Email me direct.

Thanks

Roy Beavers
Southern Methodist University
Department of Earth Sciences
P.O. Box 750395
Dallas, TX  75275
Voice: 214-768-2756
Fax: 214-768-2701
Email: rbeavers-at-smu.edu



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From: werner1-at-slb.com
Date: Wed, 28 Apr 2010 14:12:50 -0500
Subject: [Microscopy] Measurements

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Ed,

I was waiting for someone who knew what they were doing to reply to your request, but since nobody has done so - here goes.

You can download ImageJ and plugins at http://rsb.info.nih.gov/ij/download.html - read more at the home page at http://rsb.info.nih.gov/ij/ . There is an automatic update plugin that works like a charm - http://rsb.info.nih.gov/ij/plugins/imagej-updater.html .

Assuming the image on which you want to measure features already has a micron marker put there by the microscope, here is how you tell ImageJ how many pixels to the micron so you can make measurements:

Open ImageJ and open the image on which you want to measure. Click on the straight line tool, 5th from the left on the ImageJ toolbar. Draw a straight line using this tool, overlaying (overlying?) the scale bar exactly.

Click on Analyze, move down to Set Scale, and click. Opens a box with spaces for you to put "known distance" (the length of the scalebar you drew the overlay on) and Unit of length - erase "pixel" and put micron, or whatever your scale bar is. Check the box that says Global.

Now, whenever you use the straight line tool between two points on that image, the line length will show up at the bottom of the ImageJ toolbar. When I figure out how to get it to print on the image I'll post that, but don't hold your breath.

Sorry this is not more complete but I'm still an ImageJ novice.

Regards,
Andrew


-----Original Message-----
X-from: ehaller-at-health.usf.edu [mailto:ehaller-at-health.usf.edu]
Sent: Tuesday, April 27, 2010 10:32 AM
To: Andrew Werner

Hi, Ian,

I can't offer much help, I'm afraid. I'm in the same boat. I'm writing to ask that if you receive replies off the listserver that you please post them to the listserver. I have a project of 60 SEM samples of sharkskin where we need to measure dermal scale distance on tiff images. I'm aware of the arguments against accuracy of measurements from SEM images, but we're doing low mag work on basically flat images at essentially zero tilt. We don't have the photos yet. I have a couple of old calibration standards from Ladd and Fullam to calibrate the mag in my SEM. I've checked it before and the readout was accurate.
I was going to fool around with Photoshop, a program called FijiImage, which is an offshoot of ImageJ, and then try to import the tiffs into my TEM where I have Olympus Soft Imaging software (AnalySis), but that software is calibrated for my TEM mag's so, I would have a lot of work there to recalibrate it for the SEM mags before I could use its measure function. Those are the options I have open.

FijiImage can be downloaded at: http://pacific.mpi-cbg.de/wiki/index.php/Main_Page

For those ImageJ users, I've forgotten how to load and open plugins. I know there's a measure plug-in that can be downloaded and a way to calibrate ImageJ for microns. Maybe you can tell us where it is and walk Ian and I through loading and using it in a non-technical way. I'm user-manual challenged :)

Ed

Edward Haller, Lab Manager
Integrative Biology Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-cas.usf.edu

________________________________________
X-from: ian.drucker-at-gmail.com [ian.drucker-at-gmail.com]
Sent: Tuesday, April 27, 2010 9:37 AM
To: Haller, Edward

I'm looking for a free ready to use program that can do measurements
of Tif images from a SEM. I know there's several free image
manipulation packages out there but I haven't found a ready to use
measurement function that can take into account the image
magnification and such.

Anyone with some suggestions?


Thanks

==============================Original Headers==============================snipped


==============================Original Headers==============================
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29, 33 -- From: Andrew Werner {werner1-at-slb.com}
29, 33 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
29, 33 -- Date: Wed, 28 Apr 2010 21:12:46 +0200
29, 33 -- Subject: RE: [Microscopy] RE: Measurements
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From: drix00-at-gmail.com
Date: Wed, 28 Apr 2010 14:41:39 -0500
Subject: [Microscopy] Re: Measurements

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

Hi,

To almost get the value of the measurement on the image with ImageJ
you can follow these step:

- Open "ROI Manager" window: Analyze-} Tools-} ROI Manager
- Draw a line on the feature you want to measure
- Add it to the ROI manager: Click Add or press t
- You can rename the new roi by selecting it and click rename.
- Add all your lines measurement you want.
- Select the measurement you want in Analyze-} Set Measurements, you
should select Display Label and Area for the line length.
- Select all rois in the "ROI Manager" and click measure, a new
windows will show up with the measurement. You can save it in a text
file.
- Check the "Show All" option in "ROI Manager" to see all line on the image.
- To save the image with the line, Flatten the image in "ROI Manager"
and save it.

Good luck,
Hendrix

On Wed, Apr 28, 2010 at 3:16 PM, {werner1-at-slb.com} wrote:
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Ed,
}
} I was waiting for someone who knew what they were doing to reply to your request, but since nobody has done so - here goes.
}
} You can download ImageJ and plugins at http://rsb.info.nih.gov/ij/download.html - read more at the home page at http://rsb.info.nih.gov/ij/ .  There is an automatic update plugin that works like a charm - http://rsb.info.nih.gov/ij/plugins/imagej-updater.html .
}
} Assuming the image on which you want to measure features already has a micron marker put there by the microscope, here is how you tell ImageJ how many pixels to the micron so you can make measurements:
}
} Open ImageJ and open the image on which you want to measure.  Click on the straight line tool, 5th from the left on the ImageJ toolbar.  Draw a straight line using this tool, overlaying (overlying?) the scale bar exactly.
}
} Click on Analyze, move down to Set Scale, and click.  Opens a box with spaces for you to put "known distance" (the length of the scalebar you drew the overlay on) and Unit of length - erase "pixel" and put micron, or whatever your scale bar is.  Check the box that says Global.
}
} Now, whenever you use the straight line tool between two points on that image, the line length will show up at the bottom of the ImageJ toolbar.  When I figure out how to get it to print on the image I'll post that, but don't hold your breath.
}
} Sorry this is not more complete but I'm still an ImageJ novice.
}
} Regards,
} Andrew
}
}
} -----Original Message-----
} X-from: ehaller-at-health.usf.edu [mailto:ehaller-at-health.usf.edu]
} Sent: Tuesday, April 27, 2010 10:32 AM
} To: Andrew Werner
} Subject: [Microscopy] RE: Measurements
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi, Ian,
}
}     I can't offer much  help, I'm afraid. I'm in the same boat. I'm writing to ask that if you receive replies off the listserver that you please post them to the listserver. I have a project of 60 SEM samples of sharkskin where we need to measure dermal scale distance on tiff images. I'm aware of the arguments against accuracy of measurements from SEM images, but we're doing low mag work on basically flat images at essentially zero tilt. We don't have the photos yet. I have a couple of old calibration standards from Ladd and Fullam to calibrate the mag in my SEM. I've checked it before and the readout was accurate.
}     I was going to fool around with Photoshop, a program called FijiImage, which is an offshoot of ImageJ, and then try to import the tiffs into my TEM where I have Olympus Soft Imaging software (AnalySis), but that software is calibrated for my TEM mag's so, I would have a lot of work there to recalibrate it for the SEM mags before I could use its measure function. Those are the options I have open.
}
}  FijiImage can be downloaded at: http://pacific.mpi-cbg.de/wiki/index.php/Main_Page
}
} For those ImageJ users, I've forgotten how to load and open plugins. I know there's a measure plug-in that can be downloaded and a way to calibrate ImageJ for microns. Maybe you can tell us where it is and walk Ian and I through loading and using it in a non-technical way. I'm user-manual challenged :)
}
} Ed
}
} Edward Haller, Lab Manager
} Integrative Biology Electron Microscopy Core
} SCA 110
} 4202 East Fowler Avenue
} Tampa, FL 33620
} (813)974-2676
} ehaller-at-cas.usf.edu
}
} ________________________________________
} X-from: ian.drucker-at-gmail.com [ian.drucker-at-gmail.com]
} Sent: Tuesday, April 27, 2010 9:37 AM
} To: Haller, Edward
} Subject: [Microscopy] Measurements
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} I'm looking for a free ready to use program that can do measurements
} of Tif images from a SEM.   I know there's several free image
} manipulation packages out there but I haven't found a ready to use
} measurement function that can take into account the image
} magnification and such.
}
} Anyone with some suggestions?
}
}
} Thanks
}
} ==============================Original Headers==============================snipped
}
}
} ==============================Original Headers==============================
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From: Ian.Hallett-at-plantandfood.co.nz
Date: Wed, 28 Apr 2010 16:33:16 -0500
Subject: [Microscopy] Image Database

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

Our institute is investigating implementing an image database across the
whole organisation, this would cover all forms of image data,
microscopes, gel scans , studio and field photography, marketing
resources etc. The institute has over 900 staff based at 15 sites
through New Zealand. Clearly this is a somewhat larger undertaking than
the data systems we use on our individual microscopes. How well has
such a system been implemented at other similar sized organisations,
what should I be looking out for (particularly in respect to the
microscope imaging area), any suggestions of software/hardware solutions
that have worked particularly well.

Thanks

Ian

Ian Hallett
Senior Scientist
Team Leader - Microscopy and Cell Walls

Plant & Food Research

T: +64 9 925 7027
F: +64 9 925 7001
ian.hallett-at-plantandfood.co.nz
www.plantandfood.co.nz
The New Zealand Institute for Plant & Food Research Limited

Postal Address: Plant & Food Research Mt Albert
Private Bag 92169, Auckland, 1142, New Zealand
Physical Address: Plant & Food Research Mt Albert
120 Mt Albert Road, Sandringham, Auckland, 1025, New Zealand



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From: hanke-at-mee-inc.com
Date: Wed, 28 Apr 2010 17:35:51 -0500
Subject: [Microscopy] Re: Measurements

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

Ian:

Try the Zeiss AxioVision freeware version. My recollection is that it
can do simple measurements once you have input appropriate calibrations.
Download at
http://www.zeiss.de/C12567BE0045ACF1/ContentsWWWIntern/CBE917247DA02A1CC1256E0000491172

--
Larry D. Hanke, P.E.
Materials Evaluation and Engineering, Inc.
Practical Solutions Through Technology and Innovation
http://www.mee-inc.com (763) 449-8870


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From: naomi_mccallum-at-health.qld.gov.au
Date: Wed, 28 Apr 2010 18:19:39 -0500
Subject: [Microscopy] Fwd: Image Database

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

Hi Ian

Clearly there will be others out there who are more familiar with options available for your situation. We are using Stream DB Management System (Olympus) and I believe it is also used at a large organisation in Victoria, Aus. and also for telepathology in Northern Territory.

There are a number of features that attracted us including ability to use the same software for acquisition and archiving of LM images. The data security was important (as we work in diagnostic pathology) and it provides SQL/Oracle login with the facility for "Permission Sets" to limit who has access to what and what they can do with it. The rep will be able to provide more info on the encryption etc.
Many file types are supported (not just for image files) so all associated data can be archived with the images.

Our DB sits on a central server with the repository for files and access is being rolled out to multiple sites across the state. It may be possible to host the main licence at a central site with repositories at each of the sites. No doubt it will come down to your IT setup and support. It does require an administrator but this is not too imposing for someone who has a day job.

I'm happy to answer further questions (although I caution that I'm an IT novice) or put you in touch with the rep.

regards
Naomi
} } } {Ian.Hallett-at-plantandfood.co.nz} 29/04/2010 7:41 am } } }



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Our institute is investigating implementing an image database across the
whole organisation, this would cover all forms of image data,
microscopes, gel scans , studio and field photography, marketing
resources etc. The institute has over 900 staff based at 15 sites
through New Zealand. Clearly this is a somewhat larger undertaking than
the data systems we use on our individual microscopes. How well has
such a system been implemented at other similar sized organisations,
what should I be looking out for (particularly in respect to the
microscope imaging area), any suggestions of software/hardware solutions
that have worked particularly well.

Thanks

Ian

Ian Hallett
Senior Scientist
Team Leader - Microscopy and Cell Walls

Plant & Food Research

T: +64 9 925 7027
F: +64 9 925 7001
ian.hallett-at-plantandfood.co.nz
www.plantandfood.co.nz
The New Zealand Institute for Plant & Food Research Limited

Postal Address: Plant & Food Research Mt Albert
Private Bag 92169, Auckland, 1142, New Zealand
Physical Address: Plant & Food Research Mt Albert
120 Mt Albert Road, Sandringham, Auckland, 1025, New Zealand



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Naomi McCallum
Supervising Scientist, EM Unit, Anatomical Pathology
Pathology Queensland
_________________________________________________
Clinical and Statewide Services Division| Queensland Health

Block 7 Level 2
RBWH Queensland 4029
Ph: 07 3636 8057
Mob:
Fax: 07 3636 8908
Email: naomi_mccallum-at-health.qld.gov.au
Web: http://www.health.qld.gov.au/qhcss/qhps


********************************************************************************
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From: bvos-at-SCKCEN.BE
Date: Thu, 29 Apr 2010 04:38:34 -0500
Subject: [Microscopy] Conductive cold resin for SEM.

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

Dear all,

The curing time of the conductive cold resin (technovit 5000) we use, is
to short while curing already starts during mixing of both components.
As a result, the viscosity of the resulting paste is to high for our
application. Does there exist an alternative with slow curing property?

Benedict Vos

SCK/CEN
NMS/MNA
LHMA

Boeretang 200
B-2400 Mol
Belgium

Tel. : (+32-14) 33 30 65
Fax :(+32-14) 32 12 16


SCK·CEN Disclaimer: http://www.sckcen.be/en/Legal-aspects/E-mail-disclaimer




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From: curator-at-imaginationgalleries.com
Date: Thu, 29 Apr 2010 07:50:59 -0500
Subject: [Microscopy] viaWWW: Where are all the Microscopic images

Contents Retrieved from Microscopy Listserver Archives
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This Question/Comment was submitted to the Microscopy Listserver
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Email: curator-at-imaginationgalleries.com
Name: Aaron Taylor

Organization: AI

Title-Subject: [Filtered] Where are all the Microscopic images...

Message: There use to be microscopic images that were blown up. What
happened to them? It was so much fun to see images on a large scale
and guess what it was; my friends and I use to have fun with the kids
and it was a great teaching agent.

Best Regards,

Aaron

Login Host: 161.38.221.223
---------------------------------------------------------------------------

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8, 11 -- Subject: viaWWW: Where are all the Microscopic images
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From: ian.drucker-at-gmail.com
Date: Thu, 29 Apr 2010 08:14:43 -0500
Subject: [Microscopy] RE: Measurements

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

Thank You all for your help with my question!

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1, 30 -- Message-ID: {m2gc0bfda951004290614p8709544bpf650e216083e64a8-at-mail.gmail.com}
1, 30 -- Subject: RE: Measurements
1, 30 -- To: Microscopy-at-microscopy.com
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==============================End of - Headers==============================




From: mike.bode-at-resaltatech.com
Date: Thu, 29 Apr 2010 09:22:27 -0500
Subject: [Microscopy] RE: Measurements

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

Hello Ian,

I was struggling with my emails and the listserver, so, perhaps a bit late,
but here is another possibility:

We have a free version of our Scandium software (called Scandium MeasureIT)
which can do some manual measurements, and will keep track of calibration
and magnification, even prints a scale bar if you want it to. Let me know if
you are interested, and I can send it to you. You can find out more here:
http://www.resaltatech.com/scandium_measureit_main.htm

Good luck.

Mike Bode

---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com






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From: mike.bode-at-resaltatech.com
Date: Thu, 29 Apr 2010 11:19:51 -0500
Subject: [Microscopy] Apologies for server down

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

My apologies to everybody who tried to access our server or ftp server
today. I had sent out a message to the list server with an URL for more
information about the Scandium MeasureIT software. Unfortunately, I found
out this morning that the site we are using to host the website has a server
problem and the ResAltaTech.com site is down at the moment, and a generic
page is displayed. We are working to correct this.

Mike


---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com






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From: mpaetkau-at-tru.ca
Date: Fri, 30 Apr 2010 11:42:46 -0500
Subject: [Microscopy] viaWWW: Serial Communication to Hitatchi7600

Contents Retrieved from Microscopy Listserver Archives
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Email: mpaetkau-at-tru.ca
Name: Mark Paetkau

Organization: Thompson Rivers University

Title-Subject: [Filtered] Serial Communication to Hitatchi7600

Message: We currently have an Zeiss EM10 with a CCD camera
(Hammamatsu). The camera is controlled by software from AMT and the
software is set up for a serial port connection to a Hitatchi7600
electron microscope. We would like to interface the EM10 with the
software and therefore require the serial port protocol for the
Hitatchi 7600. I believe it sends magnification info as well as
position, voltage etc. We need to know the data it sends and the
format so we can emulate the Hitatchi serial port and the software
will recognize the data.

So, ideally, a copy of the Hitatchi manual dealing with the serial
port protocols would be ideal...but any info from experienced persons
would be great.

Thanks

Mark

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From: elidre-at-gmail.com
Date: Fri, 30 Apr 2010 11:43:13 -0500
Subject: [Microscopy] viaWWW: TEM of aggrecan cartilage proteoglycan

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Email: elidre-at-gmail.com
Name: Elizabeth Dreher

Organization: Drexel University

Title-Subject: [Filtered] TEM of aggrecan cartilage proteoglycan

Message: I am interested in obtaining TEM images of the aggregate
proteoglycan, aggrecan. I have lyophilized aggrecan that was
purchased in 2006, and I believe it expired in 2008. It has been
stored at -20 C the entire time, so I am wondering to what extent the
aggrecan molecules might have degraded from storage alone.

Moreover, the aggrecan has been reconstituted in deionized distilled
water to 1 mg/ml, then stored back at - 20 C. I am wondering to what
extent 2 freeze-thaw cycles might affect the aggrecan molecule.

To prepare the aggrecan for TEM, a 500 ug/ml solution in DI water was
prepared. A drop of this aggrecan solution was placed onto 400 mesh
copper TEM grid and allowed to sit for 5 minutes. Then the grid was
touched to bibulous paper to wick away the aggrecan solution. The
grid was then stained in uranyl acetate in water [1 mM] for 10
seconds by placing the grid onto a drop of uranyl acetate. Then the
grid was placed on a few drops of water to rinse, after which excess
water was wicked off the grid with bibulous paper. The grids were
allowed to air dry.

TEM imaging was performed at 80 keV or 120 keV. We have not been able
to find aggrecan molecules in our images, and are looking for any
advice on the sample prep or the TEM imaging of aggrecan. Any help
would be greatly appreciated. Thanks in advance!

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From: stefan.diller-at-t-online.de
Date: Fri, 30 Apr 2010 11:57:12 -0500
Subject: [Microscopy] Re: viaWWW: Serial Communication to Hitatchi7600

Contents Retrieved from Microscopy Listserver Archives
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Hi Mark,
the EM10 normally has no serial port. You can add this function by "grabbing" the position of the magnification switch and the
high voltage with this Interface:
http://www.stefan-diller.com/Medien/html/rem_tem4_en.htm
It should be attached to your EM10 by a service technician or somebody, who can carefully do some soldering connections :-)

Best regaards,
Stefan



Am 30.04.10 18:49, schrieb mpaetkau-at-tru.ca:
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} Email: mpaetkau-at-tru.ca
} Name: Mark Paetkau
}
} Organization: Thompson Rivers University
}
} Title-Subject: [Filtered] Serial Communication to Hitatchi7600
}
} Message: We currently have an Zeiss EM10 with a CCD camera
} (Hammamatsu). The camera is controlled by software from AMT and the
} software is set up for a serial port connection to a Hitatchi7600
} electron microscope. We would like to interface the EM10 with the
} software and therefore require the serial port protocol for the
} Hitatchi 7600. I believe it sends magnification info as well as
} position, voltage etc. We need to know the data it sends and the
} format so we can emulate the Hitatchi serial port and the software
} will recognize the data.
}
} So, ideally, a copy of the Hitatchi manual dealing with the serial
} port protocols would be ideal...but any info from experienced persons
} would be great.
}
} Thanks
}
} Mark
}
} Login Host: 206.123.162.220
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--
-----------------------------------------------------
Stefan Diller - Wissenschaftliche Photographie
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

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From: drk-at-shcc.org
Date: Fri, 30 Apr 2010 17:04:17 -0500
Subject: [Microscopy] fixation and embedding of liver for TEM

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Hello All,


We have had consistent problems preparing liver for TEM. The tissue blocks are extremely brittle and nearly impossible to cut good thick sections for LM or ultrathins for TEM. There must be a trick specific for liver that we am not aware of. Would someone who processes liver for TEM please suggest a modification to our protocol? We fix in 1.5 % glut/1.5% phosphate buffered paraformaldehyde containing 0.05% tannic acid for several hours, rinse, postfix in 1% OsO4 then dehydrate to 100% EtOH, rinse in propylene oxide, then infiltrate and embed in Spurrs epoxy and polymerize at 60-70 C overnight.

Thank you!

Doug

Douglas R. Keene
Assistant Investigator
Micro-Imaging Center
Shriners Hospital for Children
3101 S.W. Sam Jackson Park Road
Portland, Oregon 97239
503-221-3434
drk-at-shcc.org


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From: bozzola-at-siu.edu
Date: Fri, 30 Apr 2010 17:15:02 -0500
Subject: [Microscopy] Re: fixation and embedding of liver for TEM

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Hello Doug,

Could there be traces of water in the specimen or the resin? Spurrs
becomes brittle (shatters like glass when you attempt to trim it with
a razor blade) when traces of water are present. Thick sections would
be very difficult under such conditions.

I haven't cut liver in many years, but I do recall that is is not one
of the easier tissues to cut and tends to be crumbly or brittle,
anyways. This would be exacerbated if residual water is present.

Do you have any other embedments available, like Epon (or relatives)?

Let us know what solves the problem.

JB

On Fri, Apr 30, 2010 at 5:05 PM, {drk-at-shcc.org} wrote:
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}
}
} We have had consistent problems preparing liver for TEM.  The tissue blocks are extremely brittle and nearly impossible to cut good thick sections for LM or ultrathins for TEM.  There must be a trick specific for liver that we am not aware of.  Would someone who processes liver for TEM please suggest a modification to our protocol?  We fix in 1.5 % glut/1.5% phosphate buffered paraformaldehyde containing 0.05% tannic acid for several hours, rinse, postfix in 1% OsO4 then dehydrate to 100% EtOH, rinse in propylene oxide, then infiltrate and embed in Spurrs epoxy and polymerize at 60-70 C overnight.
}
} Thank you!
}
} Doug
}
} Douglas R. Keene
} Assistant Investigator
} Micro-Imaging Center
} Shriners Hospital for Children
} 3101 S.W. Sam Jackson Park Road
} Portland, Oregon  97239
} 503-221-3434
} drk-at-shcc.org
}
}
} ==============================Original Headers==============================
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} 7, 23 -- Date: Fri, 30 Apr 2010 15:08:34 -0700
} 7, 23 -- Subject: fixation and embedding of liver for TEM
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--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
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Phone: 618-453-3730


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From: renaudgeological-at-execulink.com
Date: Fri, 30 Apr 2010 17:41:19 -0500
Subject: [Microscopy] willemite and hemimorphite

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

Hello All,

The reason I am writing is that I am currently working on a project
involving willemite and hemimorphite and would like to ask a question. The
rocks I am working on supposedly only contain hemimorphite, however, I have
come across what I believe to be "willemite". I analyzed the hemimorphite
and achieved a total of ~92-93 wt% (66 wt% ZnO and 26 wt% SiO2) which allows
for 7-8wt% water. I analyzed the supposed "willemite" and I get totals of
~100 wt% (73 wt% ZnO and 27 wt% SiO2). As a confirmation, I submitted the
grain for micro-xrd and the structure of the "willemite" came back as the
hemimorphite structure. So it turns out I have several grains which mineral
chemically resemble "willemite" but structurally resemble hemimorphite.
Have you seen this before and can you recommend any papers which may relate
to this.

Thanks in advance.

Jim.


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From: dac-at-research.umass.edu
Date: Sat, 1 May 2010 13:17:54 -0500
Subject: [Microscopy] Re: fixation and embedding of liver for TEM

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Hi Doug,

the "Spurr's Resin" caught my eye. I don't know what vintage of resin
components you are using but that might be a problem. The kits were
reformulated a few years back when VCD was no longer available (at least
for this purpose) and the properties of the replacement component
ERL-4221 is different - much higher viscosity, but more importantly,
chemically different, requiring a different balance of components. E.
Ann Ellis published on this back in 2006. However some vendors shipped
kits called "Spurr's Resin" with the new component and still the
original formulas on the included datasheet and no mention of the
change; in that form it is NOT "Spurr's". I had a spell of resin
problems with "Spurr's" until some people on this list clued me in. The
blocks were miserable. With the Ellis info the blocks are super. I use
the low viscosity variant with Quetol. I am including the basics below,
but can send all the info I collected separately if you are interested.

Hope this helps. (Sorry, this may not format properly...)

Dale

Spurr-replacement - "Spurr - New Formulation" -

E. Ann Ellis, Microscopy Today, Vol 14, No 4, July 2006

Microscopy Today, July 2006, E. Ann Ellis "Solutions to the
Problem of Substitution of ERL 4221 for Vinyl Cyclohexene Dioxide in
Spurr Low Viscosity Embedding Formulations"

All formulas are for a "10g batch" - with reference to (epoxy + acid
anhydride)

Epoxy Anhyride Catalyst Comments
-------- -------- --------- -------- --------

ERL 4221 NSA DER-736 DMAE weights are in grams

4.10 5.90 0.95 0.10 Hard
4.10 5.90 1.43 0.10 Standard
4.10 5.90 1.90 0.10 Soft


Low Viscosity Formula (Half the viscosity of the "Refomulated Spurr's")
-------------------------
ERL 4221 2.22g
Quetol-651 1.40g
NSA 6.38g
DER-736 1.43g
BDMA 0.2g

Comments: Some think BDMA should be less, 0.1g.
I (DAC) used 0.09g DMAE (5 drops from a Samco #241 pipet to avoid
weighing...) and it set nicely in 16h -at- 70C


drk-at-shcc.org wrote:
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} We have had consistent problems preparing liver for TEM. The tissue blocks are extremely brittle and nearly impossible to cut good thick sections for LM or ultrathins for TEM. There must be a trick specific for liver that we am not aware of. Would someone who processes liver for TEM please suggest a modification to our protocol? We fix in 1.5 % glut/1.5% phosphate buffered paraformaldehyde containing 0.05% tannic acid for several hours, rinse, postfix in 1% OsO4 then dehydrate to 100% EtOH, rinse in propylene oxide, then infiltrate and embed in Spurrs epoxy and polymerize at 60-70 C overnight.
}
} Thank you!
}
} Doug
}
} Douglas R. Keene
} Assistant Investigator
} Micro-Imaging Center
} Shriners Hospital for Children
} 3101 S.W. Sam Jackson Park Road
} Portland, Oregon 97239
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} drk-at-shcc.org
}
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From: nizets2-at-yahoo.com
Date: Mon, 3 May 2010 06:00:57 -0500
Subject: [Microscopy] viaWWW: TEM of aggrecan cartilage proteoglycan

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Hi Elizabeth!

Not being a proteoglycan fanatic, I have no idea how big these aggregans can be. Do you have any idea yourself?
Having an idea of the size of an object really help when one has to think about a strategy in microscopy.

Now for the technical side of things: I wonder if you were precise enough with the description of your protocol.
For example, you say that you place a drop onto a copper grid. Well, if your grid is not coated, you probably know that they are full of holes right?
Given that holes are not ideal surface to dry on, your sample will dry not on the holes, but on the grid bars, so you cannot see them!
So first advice: use formvar/carbon-coated grids.

Now when one wants to immobilize a protein, it is often useful to coat the surface with poly L-lysine.
You may want to try.
I have heard that glow-discharging the grids can also help the protein to stick on the surface.

Now for the contrast, it depends on the size of the aggregates. Negative staining (with Uranyle acetate or other salts) is a good idea. I am not sure whether you need to rince after staining though.
You may also want to try shadowing.

As for the storage at -20°C, as I understood it already happened and you cannot turn back time (if you can please contact me I am interested), so you'll probably have to live with it.

Good luck with your project,

Stephane


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Email: elidre-at-gmail.com
Name: Elizabeth Dreher

Organization: Drexel University

Title-Subject: [Filtered] TEM of aggrecan cartilage proteoglycan

Message: I am interested in obtaining TEM images of the aggregate
proteoglycan, aggrecan. I have lyophilized aggrecan that was
purchased in 2006, and I believe it expired in 2008. It has been
stored at -20 C the entire time, so I am wondering to what extent the
aggrecan molecules might have degraded from storage alone.

Moreover, the aggrecan has been reconstituted in deionized distilled
water to 1 mg/ml, then stored back at - 20 C. I am wondering to what
extent 2 freeze-thaw cycles might affect the aggrecan molecule.

To prepare the aggrecan for TEM, a 500 ug/ml solution in DI water was
prepared. A drop of this aggrecan solution was placed onto 400 mesh
copper TEM grid and allowed to sit for 5 minutes. Then the grid was
touched to bibulous paper to wick away the aggrecan solution. The
grid was then stained in uranyl acetate in water [1 mM] for 10
seconds by placing the grid onto a drop of uranyl acetate. Then the
grid was placed on a few drops of water to rinse, after which excess
water was wicked off the grid with bibulous paper. The grids were
allowed to air dry.

TEM imaging was performed at 80 keV or 120 keV. We have not been able
to find aggrecan molecules in our images, and are looking for any
advice on the sample prep or the TEM imaging of aggrecan. Any help
would be greatly appreciated. Thanks in advance!

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From: drk-at-shcc.org
Date: Mon, 3 May 2010 11:25:12 -0500
Subject: [Microscopy] fixation of liver, continued

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

Hello All,

Thank you for those many that have tried to help with my problems with fixation and embedding of liver samples. There have been many good suggestions but so far none solves my problem.

I would like to be a bit more specific in describing our protocol just in case someone might think of another possible cause.

To recap, we have had consistent problems with extremely brittle blocks when cutting thick or ultrathin sections of liver. The tissue (not the surrounding blank epon) fractures like glass, even during trimming with a double edge razor blade. We use the same protocol we use for the many other tissues that we embed well, so this problem is specific to liver. Our protocol includes primary phosphate buffered fixative (1.5% glut/1.5% parform with 0.05% tannic acid) followed by 1% OsO4 (with appropriate buffer rinses), dehydration in EtOH to 100%, a rinse in PO and infiltration and embedding in Spurrs resin, mixed according to Ellis. We have also embedded in EPON 812 substitute with similar brittleness. As I say, we use this protocol successfully for all other tissues.

One suggestion received is that there is some water left in the sample which makes embedded tissue in Spurrs very brittle. I am sure our ethanol, PO and media is dry, but is it necessary to dehydrate liver longer than other tissues?

The tissues are initially fixed in another laboratory, then sent to us. The tissue pieces are about 2mm x 2mm but were fixed in two changes of primary fixative for several hours each; then stored for a couple of weeks, then sent to us. Once in my laboratory, they are cut down so that one face is no larger than 0.5mm. Then they are osmicated, dehydrated and embedded. Could either the initial excessive size of the tissues, excessive fixation time or storage in buffer be a factor?

Does anyone have another suggestion? If you work successfully with liver, would you mind sending a protocol?

Many thanks,

Doug

Douglas R. Keene
Assistant Investigator
Micro-Imaging Center
Shriners Hospital for Children
3101 S.W. Sam Jackson Park Road
Portland, Oregon 97239
drk-at-shcc.org



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12, 23 -- Date: Mon, 3 May 2010 09:29:31 -0700
12, 23 -- Subject: fixation of liver, continued
12, 23 -- Thread-Topic: fixation of liver, continued
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From: joelsheffield-at-gmail.com
Date: Mon, 3 May 2010 12:00:59 -0500
Subject: [Microscopy] Re: fixation of liver, continued

Contents Retrieved from Microscopy Listserver Archives
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It seems to me as if the problem is in the embedding, and not the tissue. So, several questions.

1. Is is a matter of the plastic? If you trim the back end of the block, i.e. the region away from the
tissue, is that end still brittle?

2. Is it in the deyhdration? How are you sure that your dehydration liquids are truly dry? When I
did this, many years ago, we kept molecular sieve pellets in the PO and Absolute Ethanol.

3. Overall: you have mentioned that other tissues do not give this result. Have you processed
other tissues at the same time, with the same reagents?

Best of luck
On 3 May 2010 at 11:33, drk-at-shcc.org wrote:

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} Hello All,
}
} Thank you for those many that have tried to help with my problems with fixation and embedding of liver samples. There have been many good suggestions but so far none solves my problem.
}
} I would like to be a bit more specific in describing our protocol just in case someone might think of another possible cause.
}
} To recap, we have had consistent problems with extremely brittle blocks when cutting thick or ultrathin sections of liver. The tissue (not the surrounding blank epon) fractures like glass, even during trimming with a double edge razor blade. We use the same protocol we use for the many other tissues that we embed well, so this problem is specific to liver. Our protocol includes primary phosphate buffered fixative (1.5% glut/1.5% parform with 0.05% tannic acid) followed by 1% OsO4 (with appropriate buffer rinses), dehydration in EtOH to 100%, a rinse in PO and infiltration and embedding in Spurrs resin, mixed according to Ellis. We have also embedded in EPON 812 substitute with similar brittleness. As I say, we use this protocol successfully for all other tissues.
}
} One suggestion received is that there is some water left in the sample which makes embedded tissue in Spurrs very brittle. I am sure our ethanol, PO and media is dry, but is it necessary to dehydrate liver longer than other tissues?
}
} The tissues are initially fixed in another laboratory, then sent to us. The tissue pieces are about 2mm x 2mm but were fixed in two changes of primary fixative for several hours each; then stored for a couple of weeks, then sent to us. Once in my laboratory, they are cut down so that one face is no larger than 0.5mm. Then they are osmicated, dehydrated and embedded. Could either the initial excessive size of the tissues, excessive fixation time or storage in buffer be a factor?
}
} Does anyone have another suggestion? If you work successfully with liver, would you mind sending a protocol?
}
} Many thanks,
}
} Doug
}
} Douglas R. Keene
} Assistant Investigator
} Micro-Imaging Center
} Shriners Hospital for Children
} 3101 S.W. Sam Jackson Park Road
} Portland, Oregon 97239
} drk-at-shcc.org
}
}
}
} ==============================Original Headers==============================
} 12, 23 -- From drk-at-shcc.org Mon May 3 11:25:12 2010
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--
Joel B. Sheffield, Ph.D.
Biology Department, Temple University
1900 North 12th Street
Philadelphia, PA 19122
jbs-at-temple.edu
(215) 204 8839, fax (215) 204 0486
http://astro.temple.edu/~jbs


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From: rosemary.white-at-csiro.au
Date: Mon, 3 May 2010 17:24:29 -0500
Subject: [Microscopy] fixation of liver, continued

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

Well, I hesitated to respond, but when we see this problem in plant tissues,
it's the embedding - resin infiltration, that's the problem, not fixation or
dehydration. You may want to try infiltrating more slowly - with smaller
resin increments, or for somewhat longer than whatever protocol you're using
at present. In plants, the problem is almost impermeable cell wall (i.e.
extracellular matrix) layers in the tissue that either prevent or greatly
slow down infiltration of resin polymers. Are there any structures in liver
that might slow polymer diffusion?

In algae, for example, some ECM components are very impermeable, especially
after osmication, and to avoid collapsing algal zygotes in some species
(because solvent can diffuse out rapidly but resin can diffuse in only
slowly), a colleague used to increment the resin by 2% per day over the
beginning and end 10-16% steps of infiltration, with larger steps in the
intermediate concentrations - it took a month to infiltrate. It did mean
she had to do this only once and had perfect TEMs.... That's an extreme
example, of course!
good luck,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au



On 4/05/10 3:05 AM, "joelsheffield-at-gmail.com" {joelsheffield-at-gmail.com}
wrote:

}
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} It seems to me as if the problem is in the embedding, and not the tissue. So,
} several questions.
}
} 1. Is is a matter of the plastic? If you trim the back end of the block,
} i.e. the region away from the
} tissue, is that end still brittle?
}
} 2. Is it in the deyhdration? How are you sure that your dehydration liquids
} are truly dry? When I
} did this, many years ago, we kept molecular sieve pellets in the PO and
} Absolute Ethanol.
}
} 3. Overall: you have mentioned that other tissues do not give this result.
} Have you processed
} other tissues at the same time, with the same reagents?
}
} Best of luck
} On 3 May 2010 at 11:33, drk-at-shcc.org wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} } ----------------------------------------------------------------------------
} }
} } Hello All,
} }
} } Thank you for those many that have tried to help with my problems with
} } fixation and embedding of liver samples. There have been many good
} } suggestions but so far none solves my problem.
} }
} } I would like to be a bit more specific in describing our protocol just in
} } case someone might think of another possible cause.
} }
} } To recap, we have had consistent problems with extremely brittle blocks when
} } cutting thick or ultrathin sections of liver. The tissue (not the
} } surrounding blank epon) fractures like glass, even during trimming with a
} } double edge razor blade. We use the same protocol we use for the many other
} } tissues that we embed well, so this problem is specific to liver. Our
} } protocol includes primary phosphate buffered fixative (1.5% glut/1.5% parform
} } with 0.05% tannic acid) followed by 1% OsO4 (with appropriate buffer rinses),
} } dehydration in EtOH to 100%, a rinse in PO and infiltration and embedding in
} } Spurrs resin, mixed according to Ellis. We have also embedded in EPON 812
} } substitute with similar brittleness. As I say, we use this protocol
} } successfully for all other tissues.
} }
} } One suggestion received is that there is some water left in the sample which
} } makes embedded tissue in Spurrs very brittle. I am sure our ethanol, PO and
} } media is dry, but is it necessary to dehydrate liver longer than other
} } tissues?
} }
} } The tissues are initially fixed in another laboratory, then sent to us. The
} } tissue pieces are about 2mm x 2mm but were fixed in two changes of primary
} } fixative for several hours each; then stored for a couple of weeks, then sent
} } to us. Once in my laboratory, they are cut down so that one face is no
} } larger than 0.5mm. Then they are osmicated, dehydrated and embedded. Could
} } either the initial excessive size of the tissues, excessive fixation time or
} } storage in buffer be a factor?
} }
} } Does anyone have another suggestion? If you work successfully with liver,
} } would you mind sending a protocol?
} }
} } Many thanks,
} }
} } Doug
} }
} } Douglas R. Keene
} } Assistant Investigator
} } Micro-Imaging Center
} } Shriners Hospital for Children
} } 3101 S.W. Sam Jackson Park Road
} } Portland, Oregon 97239
} } drk-at-shcc.org
} }
} }
} }
} } ==============================Original Headers==============================
} } 12, 23 -- From drk-at-shcc.org Mon May 3 11:25:12 2010
} } 12, 23 -- Received: from webmail.shcc.org (webmail.shcc.org [64.213.211.212])
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} } 12, 23 -- From: Doug Keene {drk-at-shcc.org}
} } 12, 23 -- To: "Microscopy-at-Microscopy.Com" {Microscopy-at-Microscopy.Com}
} } 12, 23 -- Date: Mon, 3 May 2010 09:29:31 -0700
} } 12, 23 -- Subject: fixation of liver, continued
} } 12, 23 -- Thread-Topic: fixation of liver, continued
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}
}
} --
} Joel B. Sheffield, Ph.D.
} Biology Department, Temple University
} 1900 North 12th Street
} Philadelphia, PA 19122
} jbs-at-temple.edu
} (215) 204 8839, fax (215) 204 0486
} http://astro.temple.edu/~jbs
}
}
} ==============================Original Headers==============================
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} 9, 42 -- From: joelsheffield-at-gmail.com
} 9, 42 -- To: Microscopy-at-microscopy.com, drk-at-shcc.org
} 9, 42 -- Date: Mon, 03 May 2010 13:00:43 -0400
} 9, 42 -- MIME-Version: 1.0
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==============================Original Headers==============================
10, 40 -- From prvs=73212c635=Rosemary.White-at-csiro.au Mon May 3 17:24:28 2010
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10, 40 -- Subject: Re: [Microscopy] Re: fixation of liver, continued
10, 40 -- From: Rosemary White {rosemary.white-at-csiro.au}
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From: vladislav_speransky-at-nih.gov
Date: Tue, 4 May 2010 10:30:39 -0500
Subject: [Microscopy] Re: fixation of liver, continued

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

Hi Doug!..

I also hesitated, knowing that it is unlikely that I might have any
experience beyond that of yours or John's. Since you explained more,
though, this has moved into a Mystery category, so...

(1) I do have some successful experience processing liver. It was ~15
years ago, but I remember the protocol firmly and also that there were
no trouble with blocks. I studied mitochondria as an undergraduate,
and so the ultrastructure was good too. 5% GA in 0.1 M Na PB, 2-3 h.
Bigger pieces were put in this fix and cut down to EM size by slicing
with 2 sharp razor blades. After some short time, the sample was put
in the fridge, and all steps from that point on done in the fridge,
until acetone. 1% OsO4 in same buffer. Buffer washes. 50% Ethanol,
60%, then overnight in 70% with 1.5% UA. Next day, 80%, 90%, 2x96%
(that's what it said on the bottle, pure ethanol but not dried). Then
2xacetone, 15-30' each at RT. Then 3:1, 1:1, 1:3 aceton:Epon (original
Fluka Epon). Then the barbaric step of 12-24 h at 37C in pure Epon,
then 48 h at 60C. The liver came out excellent. You can see that this
protocol is not optimal in terms of infiltration, and it did get me in
trouble with some other objects (like holes in bacteria), but not with
liver. Higher % of GA (4-5%) has always served me very well, BTW. For
the kind of work I was doing then, I never saw any advantage adding FA
to GA. Fine detail of the mitochondria always came out worse.

(2) Doug, is that human liver?.. What I did was mouse and rat of
course. With humans, who knows what they'd been eating and drinking...

(3) One thing that immediately caught my attention in your second post
- that someone else was doing the fixation. Could it be something they
do there?..

(4) Finally, if you can't solve it but still must trim and cut it, may
I recommend my favorite blades? It is # 71930 from EMS catalog,
Solingen Long Blades. Much harder and sharper than anything else.
These will make a cleaner cut.

Vlad

________________________________________________
Vlad Speransky, Staff Scientist
Supramolecular Structure and Function Resource
National Institute of Biomedical Imaging and Bioengineering, NIH
13 South Dr, Rm. 3N17 MSC 5766
Bethesda, MD 20892
301 496-3989
vladislav_speransky-at-nih.gov
http://www.nibib.nih.gov/Research/Intramural/lbps/staff/speransky

Opinions and experiences related are those of Vlad Speransky and do
not represent the NIH. On the good side, this message is not
confidential and can be freely shared and reproduced.

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

Well, I hesitated to respond, but when we see this problem in plant
tissues,
it's the embedding - resin infiltration, that's the problem, not
fixation or
dehydration. You may want to try infiltrating more slowly - with
smaller
resin increments, or for somewhat longer than whatever protocol you're
using
at present. In plants, the problem is almost impermeable cell wall
(i.e.
extracellular matrix) layers in the tissue that either prevent or
greatly
slow down infiltration of resin polymers. Are there any structures in
liver
that might slow polymer diffusion?

In algae, for example, some ECM components are very impermeable,
especially
after osmication, and to avoid collapsing algal zygotes in some species
(because solvent can diffuse out rapidly but resin can diffuse in only
slowly), a colleague used to increment the resin by 2% per day over the
beginning and end 10-16% steps of infiltration, with larger steps in the
intermediate concentrations - it took a month to infiltrate. It did
mean
she had to do this only once and had perfect TEMs.... That's an extreme
example, of course!
good luck,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au



On 4/05/10 3:05 AM, "joelsheffield-at-gmail.com" {joelsheffield-at-gmail.com}
wrote:

}
}
}
}
----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
America
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}
----------------------------------------------------------------------------
}
} It seems to me as if the problem is in the embedding, and not the
tissue. So,
} several questions.
}
} 1. Is is a matter of the plastic? If you trim the back end of the
block,
} i.e. the region away from the
} tissue, is that end still brittle?
}
} 2. Is it in the deyhdration? How are you sure that your
dehydration liquids
} are truly dry? When I
} did this, many years ago, we kept molecular sieve pellets in the PO
and
} Absolute Ethanol.
}
} 3. Overall: you have mentioned that other tissues do not give this
result.
} Have you processed
} other tissues at the same time, with the same reagents?
}
} Best of luck
} On 3 May 2010 at 11:33, drk-at-shcc.org wrote:
}
} }
} }
} }
} }
----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
America
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} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} }
----------------------------------------------------------------------------
} }
} } Hello All,
} }
} } Thank you for those many that have tried to help with my problems
with
} } fixation and embedding of liver samples. There have been many good
} } suggestions but so far none solves my problem.
} }
} } I would like to be a bit more specific in describing our protocol
just in
} } case someone might think of another possible cause.
} }
} } To recap, we have had consistent problems with extremely brittle
blocks when
} } cutting thick or ultrathin sections of liver. The tissue (not the
} } surrounding blank epon) fractures like glass, even during trimming
with a
} } double edge razor blade. We use the same protocol we use for the
many other
} } tissues that we embed well, so this problem is specific to liver.
Our
} } protocol includes primary phosphate buffered fixative (1.5% glut/
1.5% parform
} } with 0.05% tannic acid) followed by 1% OsO4 (with appropriate
buffer rinses),
} } dehydration in EtOH to 100%, a rinse in PO and infiltration and
embedding in
} } Spurrs resin, mixed according to Ellis. We have also embedded in
EPON 812
} } substitute with similar brittleness. As I say, we use this protocol
} } successfully for all other tissues.
} }
} } One suggestion received is that there is some water left in the
sample which
} } makes embedded tissue in Spurrs very brittle. I am sure our
ethanol, PO and
} } media is dry, but is it necessary to dehydrate liver longer than
other
} } tissues?
} }
} } The tissues are initially fixed in another laboratory, then sent
to us. The
} } tissue pieces are about 2mm x 2mm but were fixed in two changes of
primary
} } fixative for several hours each; then stored for a couple of
weeks, then sent
} } to us. Once in my laboratory, they are cut down so that one face
is no
} } larger than 0.5mm. Then they are osmicated, dehydrated and
embedded. Could
} } either the initial excessive size of the tissues, excessive
fixation time or
} } storage in buffer be a factor?
} }
} } Does anyone have another suggestion? If you work successfully
with liver,
} } would you mind sending a protocol?
} }
} } Many thanks,
} }
} } Doug
} }
} } Douglas R. Keene
} } Assistant Investigator
} } Micro-Imaging Center
} } Shriners Hospital for Children
} } 3101 S.W. Sam Jackson Park Road
} } Portland, Oregon 97239
} } drk-at-shcc.org
} }
} }
} }
} } ==============================Original
Headers==============================
} } 12, 23 -- From drk-at-shcc.org Mon May 3 11:25:12 2010
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} } 12, 23 -- Subject: fixation of liver, continued
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}
}
} --
} Joel B. Sheffield, Ph.D.
} Biology Department, Temple University
} 1900 North 12th Street
} Philadelphia, PA 19122
} jbs-at-temple.edu
} (215) 204 8839, fax (215) 204 0486
} http://astro.temple.edu/~jbs
}


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From: dac-at-research.umass.edu
Date: Tue, 4 May 2010 12:02:11 -0500
Subject: [Microscopy] Seeking information about sectioning crystals

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

Hello All,

I mostly work with biological samples. I have a researcher who is
working with crystals formed from ~12nm Si microspheres; this is
somewhat like opal, I think. If forms nice crystals that are easily
cleaved with a little pressure from forceps, etc. Lacking a field
emission SEM, we have shadowed the surface of the crystals and find a
nice hexagonal packing. They are a bit to small to be able to cleave in
a controlled way in a chosen plane. The researcher would like to know if
the crystals have the same order inside as we see at the surface. I
don't think it would make a crystal if it were not similarly ordered
inside, but I guess one has to look. (Get to the point, lad!) So they
would like to cut sections to see an arbitrary surface. I embedded some
crystals in hard epoxy and tried sectioning with a diamond knife; the
crystal shatters and the section separates where the crystal was,
leaving small shattered fragments clinging to the edges of the resin. I
have to admit that I did not use my good diamond knife, but an older one
that produces a few knife marks and is certainly not optimally sharp any
longer - because I didn't know if it would damage the knife. The resin
sectioned fine. I tried various cutting speeds, and thicknesses from
50nm to 90nm, all with the same outcome.

The questions...
Is it possible to section crystals in this way?
Would this likely damage a "standard" biological diamond knife.
Would FE-SEM of a cleaved crystal surface be a better approach?

Is there another approach to take?

Thanks for any ideas!

Dale Callaham



==============================Original Headers==============================
8, 20 -- From dac-at-research.umass.edu Tue May 4 12:02:10 2010
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From: kenconverse-at-qualityimages.biz
Date: Tue, 4 May 2010 12:55:45 -0500
Subject: [Microscopy] Seeking information about sectioning crystals

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

Hi Dale,
I would be inclined to try grinding and polishing rather than microtoming,
if you're using an SEM. The materials folks should be able to help you out
on that end. If you're planning on using a TEM, then you might want to
think about FIB, although I don't know if the ion beam might disturb the
crystal structure.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: dac-at-research.umass.edu [mailto:dac-at-research.umass.edu]
Sent: Tuesday, May 04, 2010 1:04 PM
To: kenconverse-at-qualityimages.biz

Hello All,

I mostly work with biological samples. I have a researcher who is
working with crystals formed from ~12nm Si microspheres; this is
somewhat like opal, I think. If forms nice crystals that are easily
cleaved with a little pressure from forceps, etc. Lacking a field
emission SEM, we have shadowed the surface of the crystals and find a
nice hexagonal packing. They are a bit to small to be able to cleave in
a controlled way in a chosen plane. The researcher would like to know if
the crystals have the same order inside as we see at the surface. I
don't think it would make a crystal if it were not similarly ordered
inside, but I guess one has to look. (Get to the point, lad!) So they
would like to cut sections to see an arbitrary surface. I embedded some
crystals in hard epoxy and tried sectioning with a diamond knife; the
crystal shatters and the section separates where the crystal was,
leaving small shattered fragments clinging to the edges of the resin. I
have to admit that I did not use my good diamond knife, but an older one
that produces a few knife marks and is certainly not optimally sharp any
longer - because I didn't know if it would damage the knife. The resin
sectioned fine. I tried various cutting speeds, and thicknesses from
50nm to 90nm, all with the same outcome.

The questions...
Is it possible to section crystals in this way?
Would this likely damage a "standard" biological diamond knife.
Would FE-SEM of a cleaved crystal surface be a better approach?

Is there another approach to take?

Thanks for any ideas!

Dale Callaham



==============================Original Headers==============================
8, 20 -- From dac-at-research.umass.edu Tue May 4 12:02:10 2010
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From: david.knecht-at-uconn.edu
Date: Tue, 4 May 2010 13:01:46 -0500
Subject: [Microscopy] Filters for TIRF

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

We have a Nikon TI with through the objective TIRF system that we have been using successfully for a year or so. As we have pushed the system harder toward single molecule imaging, we have encountered problems of interference patterns showing up in the images. We have been using a Semrock laser quad filter set, which worked much better than the standard Chroma set we started with. A dealer rep that I discussed this issue with said that the only way he got TIRF to work well, was with a new Chroma extra thick dichroic, glued into a metal cube and single emitter filters in a filter wheel. If others have put together TIRF systems, I would be interested to hear if they have encountered filter problems and what was done to solve the problems. THanks- Dave

Dr. David Knecht
Department of Molecular and Cell Biology
Co-head Flow Cytometry and Confocal Microscopy Facility
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)




==============================Original Headers==============================
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From: jerzy.gazda-at-ceriumlabs.com
Date: Tue, 4 May 2010 14:25:36 -0500
Subject: [Microscopy] Seeking information about sectioning crystals

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

Dale,
The small fragments that are attached to the sections might have the answer for you. Kind of "micro-cleaving" approach. Look at them in your TEM and evaluate the microstructure, near the sharp edges are the best locations to find thin ( {200nm for 100KV instrument) areas. Also diffraction analysis with condensed beam (CBED) might yield some suggestions on the crystallinity of the structure. Other approach would be to use "mortal and pestle", then rinse in DI water and deposit on C-coated grid, then review in bio-TEM.

Give us an update,

Jerzy

***************************************************
Jerzy Gazda Ph.D.
Section Manager - TEM, FIB, SEM
Cerium Laboratories LLC
5204 E. Ben White Blvd. MS-512
Austin, TX 78741

(512) 934-5185 vm
(512) 622-6600 pgr

www.ceriumlabs.com


-----Original Message-----
X-from: dac-at-research.umass.edu [mailto:dac-at-research.umass.edu]
Sent: Tuesday, May 04, 2010 12:11 PM
To: Gazda, Jerzy

Hello All,

I mostly work with biological samples. I have a researcher who is
working with crystals formed from ~12nm Si microspheres; this is
somewhat like opal, I think. If forms nice crystals that are easily
cleaved with a little pressure from forceps, etc. Lacking a field
emission SEM, we have shadowed the surface of the crystals and find a
nice hexagonal packing. They are a bit to small to be able to cleave in
a controlled way in a chosen plane. The researcher would like to know if
the crystals have the same order inside as we see at the surface. I
don't think it would make a crystal if it were not similarly ordered
inside, but I guess one has to look. (Get to the point, lad!) So they
would like to cut sections to see an arbitrary surface. I embedded some
crystals in hard epoxy and tried sectioning with a diamond knife; the
crystal shatters and the section separates where the crystal was,
leaving small shattered fragments clinging to the edges of the resin. I
have to admit that I did not use my good diamond knife, but an older one
that produces a few knife marks and is certainly not optimally sharp any
longer - because I didn't know if it would damage the knife. The resin
sectioned fine. I tried various cutting speeds, and thicknesses from
50nm to 90nm, all with the same outcome.

The questions...
Is it possible to section crystals in this way?
Would this likely damage a "standard" biological diamond knife.
Would FE-SEM of a cleaved crystal surface be a better approach?

Is there another approach to take?

Thanks for any ideas!

Dale Callaham



==============================Original Headers==============================
8, 20 -- From dac-at-research.umass.edu Tue May 4 12:02:10 2010
8, 20 -- Received: from race-3.oit.umass.edu (race-3.oit.umass.edu [128.119.2.108])
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==============================Original Headers==============================
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19, 29 -- Subject: RE: [Microscopy] Seeking information about sectioning crystals
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From: randerson20-at-tampabay.rr.com
Date: Tue, 4 May 2010 16:05:50 -0500
Subject: [Microscopy] Re: Seeking information about sectioning crystals

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Dear Dale,

No. You can't prepare Si specimens with a microtome.

I wrote a short note on the subject in Microscopy Today, 14, 6, November
2006, pg 58, titled "Just Say NO to Microtoming Silicon!" that explains
why you will wind up with a pile of Si dust and possibly damage your knife.

You can download an MT Archive PDF at
http://www.microscopy-today.com/index.jsf;jsessionid=15B61AFA7C53C9E9CAEAE496FEF4BA5E

Polishing and etching your mounted specimens will work, but inasmuch as
your particles will be randomly oriented, you can't count on seeing the
"nice hexagonal packing" in all of the particles. The crystal lattice is
present in all of them--it just won't be visible unless you find
particles oriented in one of the principal crystallographic
orientations. The {111} orientation for a hexagonal surface, etc.

Ron Anderson


dac-at-research.umass.edu wrote:
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} Hello All,
}
} I mostly work with biological samples. I have a researcher who is
} working with crystals formed from ~12nm Si microspheres; this is
} somewhat like opal, I think. If forms nice crystals that are easily
} cleaved with a little pressure from forceps, etc. Lacking a field
} emission SEM, we have shadowed the surface of the crystals and find a
} nice hexagonal packing. They are a bit to small to be able to cleave in
} a controlled way in a chosen plane. The researcher would like to know if
} the crystals have the same order inside as we see at the surface. I
} don't think it would make a crystal if it were not similarly ordered
} inside, but I guess one has to look. (Get to the point, lad!) So they
} would like to cut sections to see an arbitrary surface. I embedded some
} crystals in hard epoxy and tried sectioning with a diamond knife; the
} crystal shatters and the section separates where the crystal was,
} leaving small shattered fragments clinging to the edges of the resin. I
} have to admit that I did not use my good diamond knife, but an older one
} that produces a few knife marks and is certainly not optimally sharp any
} longer - because I didn't know if it would damage the knife. The resin
} sectioned fine. I tried various cutting speeds, and thicknesses from
} 50nm to 90nm, all with the same outcome.
}
} The questions...
} Is it possible to section crystals in this way?
} Would this likely damage a "standard" biological diamond knife.
} Would FE-SEM of a cleaved crystal surface be a better approach?
}
} Is there another approach to take?
}
} Thanks for any ideas!
}
} Dale Callaham
}
}
}
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Subject: [Microscopy] viaWWW: Certificate in Materials Characterization

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From: nizets2-at-yahoo.com
Date: Wed, 5 May 2010 04:08:32 -0500
Subject: [Microscopy] Seeking information about sectioning crystals

Contents Retrieved from Microscopy Listserver Archives
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I definitely second this idea.
However be careful that most often the grid is turned upside down into the microscope and if the particles deposited on it do not firmly attach, they will contamine the microscope.
So take care to shake the grid a bit to get rid of the loosely attached particles before inserting the specimen holder.

Stephane



----- Original Message ----
X-from: "jerzy.gazda-at-ceriumlabs.com" {jerzy.gazda-at-ceriumlabs.com}
To: nizets2-at-yahoo.com
Sent: Tue, May 4, 2010 9:30:16 PM

Dale,
The small fragments that are attached to the sections might have the answer for you. Kind of "micro-cleaving" approach. Look at them in your TEM and evaluate the microstructure, near the sharp edges are the best locations to find thin ( {200nm for 100KV instrument) areas. Also diffraction analysis with condensed beam (CBED) might yield some suggestions on the crystallinity of the structure. Other approach would be to use "mortal and pestle", then rinse in DI water and deposit on C-coated grid, then review in bio-TEM.

Give us an update,

Jerzy

***************************************************
Jerzy Gazda Ph.D.
Section Manager - TEM, FIB, SEM
Cerium Laboratories LLC
5204 E. Ben White Blvd. MS-512
Austin, TX 78741

(512) 934-5185 vm
(512) 622-6600 pgr

www.ceriumlabs.com


-----Original Message-----
X-from: dac-at-research.umass.edu [mailto:dac-at-research.umass.edu]
Sent: Tuesday, May 04, 2010 12:11 PM
To: Gazda, Jerzy

Hello All,

I mostly work with biological samples. I have a researcher who is
working with crystals formed from ~12nm Si microspheres; this is
somewhat like opal, I think. If forms nice crystals that are easily
cleaved with a little pressure from forceps, etc. Lacking a field
emission SEM, we have shadowed the surface of the crystals and find a
nice hexagonal packing. They are a bit to small to be able to cleave in
a controlled way in a chosen plane. The researcher would like to know if
the crystals have the same order inside as we see at the surface. I
don't think it would make a crystal if it were not similarly ordered
inside, but I guess one has to look. (Get to the point, lad!) So they
would like to cut sections to see an arbitrary surface. I embedded some
crystals in hard epoxy and tried sectioning with a diamond knife; the
crystal shatters and the section separates where the crystal was,
leaving small shattered fragments clinging to the edges of the resin. I
have to admit that I did not use my good diamond knife, but an older one
that produces a few knife marks and is certainly not optimally sharp any
longer - because I didn't know if it would damage the knife. The resin
sectioned fine. I tried various cutting speeds, and thicknesses from
50nm to 90nm, all with the same outcome.

The questions...
Is it possible to section crystals in this way?
Would this likely damage a "standard" biological diamond knife.
Would FE-SEM of a cleaved crystal surface be a better approach?

Is there another approach to take?

Thanks for any ideas!

Dale Callaham



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From: jacques.faerber-at-ipcms.u-strasbg.fr
Date: Wed, 5 May 2010 04:39:39 -0500
Subject: [Microscopy] EDS : bulk ot thin film correction

Contents Retrieved from Microscopy Listserver Archives
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Hi all

I have some samples composed most of bacteria and biofilm and more or
less mineral particules like alumino-silicates, illmenite, quartz and a
lot of salts, and I must track some As.The samples are on TEM grids and
analysed by SEM, say at 15-20 keV, mounted in way that avoid substrate
effect. I use most spot mode, soemtimes scanned squares.

What would be the size limit of mineral particules where one will switch
between a thin film correction modele or a bulk one (eg Cliff-Lorimer
versus PhiRoZ or ZAF).

Is a measurement of the transmited current a good criteria. Say, with
90% ( or 95%, 75% ???) of the primary current one use thin film
correction, and with less the bulk model ? Or would it be a other way to
do an estimation by Monte-Carlo simulation of the ratio of transmetted
electrons ?

I've looked at the classical books, but they deal either with bulk or
with thin film (in that case in a TEM at high energy, or in case of the
SEM, with film on substrate), and I found nothing about the way to
evaluate a limit to switch from one to the other. Chap 10.3 of Goldstein
and al (2003) deals with particule analysis, but more in a few tens of
µm in diameter and where the shape factor (spheres) is important. I my
case, the minerals are in a range of 0.1 to 2-5 µm, and mostly flat.

My goal is not do fine quantitative analysis, but to have some good
estimation of the concentrations.
On a reference sample with most bacteria and biofilm, less minerals and
a know concentration of As, I've soon significative differences between
the two models (overestimation of O and As in bulk correction methode).
Of coarse, I do not measure the carbon, what makes a bias too. For that
sample, the thin film model seams to be more correct.

Any advices ?

Jacques

--
J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr


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From: klivi-at-jhu.edu
Date: Wed, 5 May 2010 08:19:34 -0500
Subject: [Microscopy] Re: EDS : bulk ot thin film correction

Contents Retrieved from Microscopy Listserver Archives
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Jacques,
In my experience with TEMs and mineral analyses, it is likely that all
your samples are beyond what is called the Cliff-Lorimer thin-film
criterion. Although some researchers say that once you have absorption
of any measured X-ray by 3% by the bulk, then you are out of the thin-
film region. However, it is actually the ratio of any elements
changing by 3% that matters, since the C-L method relies on ratios.
Thus, the thin-film thickness for fayalite (Fe2SiO4) has a much
thinner thin-film thickness -- in the 10s of nm-- than forsterite
(Mg2SiO4) which is more in the µm range. In the latter case, both Mg
and Si are being absorbed nearly equally. So there is no one answer to
your question. There are two things to do: 1) Calculate the thin-film
criteria for a variety of mineral compositions that you would expect
to find. You can also simulate the effect of absorption with thickness
using the program DTSA from NIST in the USA. The original program was
compiled for old Macs. However, there is a new version being rewritten
for the PC, although I don't know the status of the rewrite. 2) If you
can trust your oxygen analyses and the minerals are stoichiometric,
then you can use the method of E van Cappellen Ultramicroscopy 53
(1994) 343 which tracks absorption by the reduction of the oxygen
analysis from stoichiometric. These are two big assumptions though.

For qualitative analyses, you can trust element ratios of neighboring
(in the periodic table) atoms, but be very suspect of ratios from X-
rays of very different energies!

A third alternative is to use TEM... my choice.

Hope this helps.

Ken

On May 5, 2010, at 5:41 AM, jacques.faerber-at-ipcms.u-strasbg.fr wrote:

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} Hi all
}
} I have some samples composed most of bacteria and biofilm and more or
} less mineral particules like alumino-silicates, illmenite, quartz
} and a
} lot of salts, and I must track some As.The samples are on TEM grids
} and
} analysed by SEM, say at 15-20 keV, mounted in way that avoid substrate
} effect. I use most spot mode, soemtimes scanned squares.
}
} What would be the size limit of mineral particules where one will
} switch
} between a thin film correction modele or a bulk one (eg Cliff-Lorimer
} versus PhiRoZ or ZAF).
}
} Is a measurement of the transmited current a good criteria. Say, with
} 90% ( or 95%, 75% ???) of the primary current one use thin film
} correction, and with less the bulk model ? Or would it be a other
} way to
} do an estimation by Monte-Carlo simulation of the ratio of transmetted
} electrons ?
}
} I've looked at the classical books, but they deal either with bulk or
} with thin film (in that case in a TEM at high energy, or in case of
} the
} SEM, with film on substrate), and I found nothing about the way to
} evaluate a limit to switch from one to the other. Chap 10.3 of
} Goldstein
} and al (2003) deals with particule analysis, but more in a few tens of
} µm in diameter and where the shape factor (spheres) is important. I my
} case, the minerals are in a range of 0.1 to 2-5 µm, and mostly flat.
}
} My goal is not do fine quantitative analysis, but to have some good
} estimation of the concentrations.
} On a reference sample with most bacteria and biofilm, less minerals
} and
} a know concentration of As, I've soon significative differences
} between
} the two models (overestimation of O and As in bulk correction
} methode).
} Of coarse, I do not measure the carbon, what makes a bias too. For
} that
} sample, the thin film model seams to be more correct.
}
} Any advices ?
}
} Jacques
}
} --
} J. Faerber
} IPCMS-GSI
} (Institut de Physique et Chimie des Matériaux de Strasbourg
} Groupe Surface et Interfaces)
} 23, rue de Loess ; BP43
} 67034 Strasbourg CEDEX 2
} France
}
} Tel 00 33(0)3 88 10 71 01
} Fax 00 33(0)3 88 10 72 48
} E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr
}
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|_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
Kenneth JT Livi, PhD
The High-Resolution Analytical Electron Microbeam Facility of the
Integrated Imaging Center
Departments of Earth and Planetary Sciences and Biology
Olin Hall
3400 N Charles Street
Johns Hopkins University
Baltimore, Maryland 21218 USA
| | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| |
| :|









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From: zaluzec-at-aaem.amc.anl.gov
Date: Wed, 5 May 2010 09:42:36 -0500
Subject: [Microscopy] Re: EDS : bulk vs thin film correction

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

Bon Jour Jacques

The criterion for "thin-film" vs "bulk" analysis should
not be based solely on thickness or simple electron transmission.

A "thin-film" analysis by (my) definition means
that the specimen thickness does not disproportionately affect
the signals being measured.

So you must evaluate the parameter you are measuring and
then determine if that parameter (in your case the intensity
of the X-ray Line) is being affected disproportionately relative
to any other signal your measuring and trying to use for
quantification.

For x-rays one typcially calculates XpT where
X=chi= massabsorption coefficient * including
geometrical factors (take of angle, sample tilt...)
p = density of the region of interest
T = sample thickness

if XpT for your ROI is { 0.1 then x-ray absorption is
neglegible and can be neglected as only a minor correction.
The important point to remember is that this is x-ray line dependant.
Thus, if your compound has a higher energy line (say Ni Ka)
it may fufill the thin-film criterion (TFC). If it also
contains low Z elements ( eg Al Ka) that line
might fail the TFC even though it satisfies the TFC for
Ni Ka , thus a correction is necessary
for all lines. I'll also add that there is also a corresponding
TFC for fluoresence corrections but it is rarely discussed.

For EELS the TFC is the T/Lambda { 1, where

T= specimen thickness
Lambda = mean free path for inelastic scattering.

Your welcome to have a look through some short course lecture
notes I have put on line on XEDS and EELS at this WWW site.

http://tpm.amc.anl.gov/Lectures/HomePage.html

Look for the XEDS short course notes.

The section you should look at is near the last quarter
of the notes.

Hope this helps, drop me a message off-line if
you have other questions. I also have a simple
Fortran program that can calculate the the TFC if you
send me your parameters and experimental
conditions (kv, geometry, thickness, nominal
compositions) I can run a few calculations for
you.

Just based upon the limited details in your posting,
I'm guessing that you will need to run a correction protocol.


Nestor
Your Friendly Neighborhood SysOp



At 4:39 AM -0500 5/5/10, jacques.faerber-at-ipcms.u-strasbg.fr wrote:
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From: nancy.hirdt-at-pepsico.com
Date: Wed, 5 May 2010 18:15:52 -0500
Subject: [Microscopy] viaWWW: thin sectioning of fruits, vegetables, seeds

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Email: nancy.hirdt-at-pepsico.com
Name: Nancy Hirdt

Organization: PepsiCo

Title-Subject: [Filtered] thin sectioning of fruits, vegetables, seeds

Message: Hello All,
I am new to the world of thin sectioning plants (fruits, vegetables,
etc) for examination with light microscopy. I would like to know
what is the difference between an ultramicrotome and a cryostat and
when would you choose one over the other.

Also, does anyone know of training that is offered to help me get
started in this area?

Best Regards,
Nancy Hirdt

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From: michelle.gignac-at-duke.edu
Date: Wed, 5 May 2010 18:16:21 -0500
Subject: [Microscopy] viaWWW: Cryo TEM Service

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Name: Michelle Gignac

Organization: Duke University

Title-Subject: [Filtered] Cryo TEM Service

Message: Hello.

Is there anyone that runs Cryo-TEM samples as a service? I have an
scientist that is interested in having one carbon nano-tube samples
imaged with Cryo-TEM.

Thanks for your help.
Michelle

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From: bfoster-at-mme1.com
Date: Wed, 5 May 2010 21:38:18 -0500
Subject: [Microscopy] Re: viaWWW: thin sectioning of fruits,

Contents Retrieved from Microscopy Listserver Archives
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Hi, Nancy

There are a number of alternatives. One thing to consider for seeds is the confocal microscope. They optically section so that you can "look inside" without having to do any cutting. We've also used this technique successfully on things like pollen.

As for the more conventional embedding and sectioning of larger items like fruits and veggies... I'll leave that to the sample prep gurus on the listserver.

Good hunting,
Barbara Foster, President and Sr. Consultant

Microscopy/Microscopy Education
7101 Royal Glen Trail, Suite A
McKinney TX 75070
P: (972)924-5310 Skype: fostermme
W: www.MicroscopyEducation.com

NEWS! Visit the NEW and IMPROVED www.MicroscopyEducation.com! And don't forget: MME is now scheduling customized, on-site courses for the Summer and Fall. Call me for a free assessment and quote.

At 06:43 PM 5/5/2010, nancy.hirdt-at-pepsico.com wrote:



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11, 21 -- Subject: Re: [Microscopy] viaWWW: thin sectioning of fruits,
11, 21 -- vegetables, seeds
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From: donald.gibbon-at-matcoinc.com
Date: Thu, 6 May 2010 15:06:15 -0500
Subject: [Microscopy] Re: viaWWW: thin sectioning of fruits,

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

Nancy: You're clearly working at a pretty strong disadvantage here. The
two things you asked about aren't even in the same category. The first
is an instrument for making thin sections, the second is a device for
keeping something very cold. Thin sections come in a huge variety of
kinds and scales, from layers of soil to bones, to oxide layers on
steels to tissue sections, from multi-centimeters to micrometers. In the
more recent decades crymicrotomes were developed to make frozen
sections... but crysostats are used many other places.

I'd suggest getting a book on the general subject of biological
microscopy (light and electron) and get an idea of the range of
approaches you could use. A good introductory course would probably be
in order too... I'd think Pepsico could afford to send you! I definitely
would NOT start with confocal microscopy if you don't yet know the
difference between a microtome and a cryostat. Confocal microscopy is
for advanced microscopists.

Donald L. Gibbon
MATCO Services

-----Original Message-----
X-from: bfoster-at-mme1.com [mailto:bfoster-at-mme1.com]
Sent: Wednesday, May 05, 2010 10:47 PM
To: Gibbon, Donald L.

Hi, Nancy

There are a number of alternatives. One thing to consider for seeds is
the confocal microscope. They optically section so that you can "look
inside" without having to do any cutting. We've also used this
technique successfully on things like pollen.

As for the more conventional embedding and sectioning of larger items
like fruits and veggies... I'll leave that to the sample prep gurus on
the listserver.

Good hunting,
Barbara Foster, President and Sr. Consultant

Microscopy/Microscopy Education
7101 Royal Glen Trail, Suite A
McKinney TX 75070
P: (972)924-5310 Skype: fostermme
W: www.MicroscopyEducation.com

NEWS! Visit the NEW and IMPROVED www.MicroscopyEducation.com! And don't
forget: MME is now scheduling customized, on-site courses for the
Summer and Fall. Call me for a free assessment and quote.

At 06:43 PM 5/5/2010, nancy.hirdt-at-pepsico.com wrote:



} -----------------------------------------------------------------------
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11, 21 -- vegetables, seeds
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From: delannoy-at-jhmi.edu
Date: Thu, 6 May 2010 15:54:27 -0500
Subject: [Microscopy] EM Microscopy Specialist

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

To the Microscopy Listserver,
We have and opening here at the Johns Hopkins School of Medicine Microscopy Facility for an Electron Microscopy Specialist. The facility has been in operation since 1989, serves over 200 users and provides Transmission, Scanning, Confocal and Multiphoton microscopy, as well as several live and light microscope stations. We have an immediate vacancy for a specialist in standard as well as cryo-ultramicrotomy, immunolabeling and cell/tissue preparation. Qualifications should include a B.S. in biology, chemistry or a related field, a minimum 3 years experience in the above mentioned techniques and strong organizational as well as interpersonal and communicative skills. The successful candidate will be working both independently and with clients who wish to be trained on various techniques and equipment. All interested parties should either contact me or HR Dept at the Johns Hopkins School of Medicine.



Michael Delannoy
Associate Director JHSM Microscope Facility
Physiology G-04
725 N Wolfe St.
Baltimore, MD 21205

(410) 955-1365


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From: c-wilke-at-northwestern.edu
Date: Thu, 6 May 2010 17:01:29 -0500
Subject: [Microscopy] UV/Low Temp Resin and Other Embedding Questions

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

Hello Listers,

I am trying to polymerize LR White at -20 C (no success after 24
hours), in a Leica AFS unit. The UV system consists of 10 UV LEDs that
are ~10 cm above the floor of the chamber. I can get the resin
polymerizes in a 60 C oven in ~12 hours. If the problem is the LR
White, can anyone advise me as to which resins work well with low
temp., UV polymerization?

Other embedding questions; I have a client that is embedding an
agarose enrobed sample in silica. Can SiO2 be sectioned? Or would one
prepare (polish) it for use in a SEM?
Another client would like to section soil by embedding in sulfur.
From what I understand, 1) the use of a cryo microtome is one method
to acquire sections.
2) and a room temperature sulfur embedding process mentions "the
plastic consistancy of the sulfur block, which is limited to 6-10
hours, before it returns to an orthorhombic state", which is not
compatible to sectioning.
Are these extremely complex processes? How does one work with sulfur?
What kinds of safety and waste disposal precautions should I be aware
of?

Thank you,
Charlene


Charlene Wilke
Imaging Specialist
Biological Imaging Facility
Northwestern University
2205 Tech Drive
Evanston, Illinois 60208
847.491.2842





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From: rosemary.white-at-csiro.au
Date: Thu, 6 May 2010 18:10:40 -0500
Subject: [Microscopy] Re: FW: Re: viaWWW: thin sectioning of fruits,

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

Hi Nancy,

I would start first with hand sections using a razor blade. If you want to
look at plants in something close to the living state, this is the way to
go. Most of the work we do here requires no more than this, if pushed we
fix and section - some cryostat sectioning, other resin sectioning, if
pushed further we go to electron microscopy.

A very good place to start is a recently published (2008) spiral-bound book
"Teaching Plant Anatomy" by RL Peterson, CA Peterson and LH Melville, NRC
Press, ISBN 978-0-660-19798-2

It's really terrific, it gives detailed step-by-step instructions and it's
all hand sectioning, staining if required, mostly simple stains, and
observing with light or fluorescence microscopy, with some macro work under
the dissector.

You can look at fruits and seeds this way, I've hand-sectioned dry barley
grains and briefly brushed the cut surface with stain and imaged under the
fluorescence dissector. We've also looked at grapes and oranges, and at the
small end I've even hand-sectioned arabidopsis anthers (a sapphire
dissecting knife is useful here). For hard or large structures, you could
investigate a sledge microtome, which people have used here to section woody
roots, hard tobacco stems, bits of trees, etc.

The only error in this book is that the microscope setup on p. 4 - "raise
and lower the condenser until the most even illumination is achieved" is not
correct, you need to adjust for Koehler illumination. The book below and
numerous websites do a better job telling you how to set up a microscope.

If you really have to, for embedded plant tissues, try "Plant Microtechnique
and Microscopy" by SE Ruzin, Oxford University Press, ISBN 0-19-508956-1

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au


On 7/05/10 6:13 AM, "donald.gibbon-at-matcoinc.com"
{donald.gibbon-at-matcoinc.com} wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Nancy: You're clearly working at a pretty strong disadvantage here. The
} two things you asked about aren't even in the same category. The first
} is an instrument for making thin sections, the second is a device for
} keeping something very cold. Thin sections come in a huge variety of
} kinds and scales, from layers of soil to bones, to oxide layers on
} steels to tissue sections, from multi-centimeters to micrometers. In the
} more recent decades crymicrotomes were developed to make frozen
} sections... but crysostats are used many other places.
}
} I'd suggest getting a book on the general subject of biological
} microscopy (light and electron) and get an idea of the range of
} approaches you could use. A good introductory course would probably be
} in order too... I'd think Pepsico could afford to send you! I definitely
} would NOT start with confocal microscopy if you don't yet know the
} difference between a microtome and a cryostat. Confocal microscopy is
} for advanced microscopists.
}
} Donald L. Gibbon
} MATCO Services
}
} -----Original Message-----
} X-from: bfoster-at-mme1.com [mailto:bfoster-at-mme1.com]
} Sent: Wednesday, May 05, 2010 10:47 PM
} To: Gibbon, Donald L.
} Subject: [Microscopy] Re: viaWWW: thin sectioning of fruits,
}
}
}
}
} ------------------------------------------------------------------------
} ----
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
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} http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ------------------------------------------------------------------------
} ----
}
} Hi, Nancy
}
} There are a number of alternatives. One thing to consider for seeds is
} the confocal microscope. They optically section so that you can "look
} inside" without having to do any cutting. We've also used this
} technique successfully on things like pollen.
}
} As for the more conventional embedding and sectioning of larger items
} like fruits and veggies... I'll leave that to the sample prep gurus on
} the listserver.
}
} Good hunting,
} Barbara Foster, President and Sr. Consultant
}
} Microscopy/Microscopy Education
} 7101 Royal Glen Trail, Suite A
} McKinney TX 75070
} P: (972)924-5310 Skype: fostermme
} W: www.MicroscopyEducation.com
}
} NEWS! Visit the NEW and IMPROVED www.MicroscopyEducation.com! And don't
} forget: MME is now scheduling customized, on-site courses for the
} Summer and Fall. Call me for a free assessment and quote.
}
} At 06:43 PM 5/5/2010, nancy.hirdt-at-pepsico.com wrote:
}
}
}
} } -----------------------------------------------------------------------
} -----
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
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} } This Question/Comment was submitted to the Microscopy Listserver
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} }
} } Email: nancy.hirdt-at-pepsico.com
} } Name: Nancy Hirdt
} }
} } Organization: PepsiCo
} }
} } Title-Subject: [Filtered] thin sectioning of fruits, vegetables, seeds
} }
} } Message: Hello All,
} } I am new to the world of thin sectioning plants (fruits, vegetables,
} } etc) for examination with light microscopy. I would like to know
} } what is the difference between an ultramicrotome and a cryostat and
} } when would you choose one over the other.
} }
} } Also, does anyone know of training that is offered to help me get
} } started in this area?
} }
} } Best Regards,
} } Nancy Hirdt
} }
} } Login Host: 198.231.24.241
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} Headers==============================
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From: rra-at-stowers.org
Date: Thu, 6 May 2010 19:48:47 -0500
Subject: [Microscopy] viaWWW: Zebra Fish Yolk

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Email: rra-at-stowers.org
Name: Rhonda Trimble

Organization: Stowers Institute

Title-Subject: [Filtered] Zebra Fish Yolk

Message: Has anyone out there ever successfully fixed and sectioned
through Zebra Fish Embryo's with the yolk? My researcher wants to
see what is going on in the yolk. I have no experience in the
matter. I am going to do some HPF, but am also looking for a
chemical fixed method newer than the 1960's.

Thanks in advance!
Rhonda Trimble

Login Host: 204.56.6.100
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From: vladislav_speransky-at-nih.gov
Date: Thu, 6 May 2010 20:55:24 -0500
Subject: [Microscopy] UV/Low Temp Resin - Why LR White??!

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

Why???

Hello Charlene,

Is it possible that you somehow have confused LR White with LR Gold?
LR Gold is the one people used to polymerize at -20C (and get endless
ribbons of sections out of it). Or Lowicryls of course. Lowicryls on
Leica AFS = tons of literature.

Sorry I don't know anything regarding your other QQs.
From my experience, one question per one post tends to work better...

Vlad
________________________________________________
Vlad Speransky, Staff Scientist
Supramolecular Structure and Function Resource
National Institute of Biomedical Imaging and Bioengineering, NIH
13 South Dr, Rm. 3N17 MSC 5766
Bethesda, MD 20892
301 496-3989
vladislav_speransky-at-nih.gov
http://www.nibib.nih.gov/Research/Intramural/lbps/staff/speransky

Opinions and experiences related are those of Vlad Speransky and do
not represent the NIH. On the good side, this message is not
confidential and can be freely shared and reproduced.

} ----------------------------------------------------------------------------
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} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hello Listers,
}
} I am trying to polymerize LR White at -20 C (no success after 24
} hours), in a Leica AFS unit. The UV system consists of 10 UV LEDs that
} are ~10 cm above the floor of the chamber. I can get the resin
} polymerizes in a 60 C oven in ~12 hours. If the problem is the LR
} White, can anyone advise me as to which resins work well with low
} temp., UV polymerization?
}
} Other embedding questions; I have a client that is embedding an
} agarose enrobed sample in silica. Can SiO2 be sectioned? Or would one
} prepare (polish) it for use in a SEM?
} Another client would like to section soil by embedding in sulfur.
} From what I understand, 1) the use of a cryo microtome is one method
} to acquire sections.
} 2) and a room temperature sulfur embedding process mentions "the
} plastic consistancy of the sulfur block, which is limited to 6-10
} hours, before it returns to an orthorhombic state", which is not
} compatible to sectioning.
} Are these extremely complex processes? How does one work with sulfur?
} What kinds of safety and waste disposal precautions should I be aware
} of?
}
} Thank you,
} Charlene
}
}
} Charlene Wilke
} Imaging Specialist
} Biological Imaging Facility
} Northwestern University
} 2205 Tech Drive
} Evanston, Illinois 60208
} 847.491.2842

==============================Original Headers==============================
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From: p.ingram-at-cellbio.duke.edu
Date: Thu, 6 May 2010 21:09:54 -0500
Subject: [Microscopy] Re: viaWWW: Zebra Fish Yolk

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

} This recent paper might be of interest to you.....

http://www.biolcell.org/boc/102/boc1020421.htm

Peter

} ---------------------------------------------------------------------------
}
} Email: rra-at-stowers.org
} Name: Rhonda Trimble
}
} Organization: Stowers Institute
}
} Title-Subject: [Filtered] Zebra Fish Yolk
}
} Message: Has anyone out there ever successfully fixed and sectioned
} through Zebra Fish Embryo's with the yolk? My researcher wants to
} see what is going on in the yolk. I have no experience in the
} matter. I am going to do some HPF, but am also looking for a
} chemical fixed method newer than the 1960's.
}
} Thanks in advance!
} Rhonda Trimble
}


--
Peter Ingram
Sr. Physicist
Adj. Professor of Pathology
Duke University Medical Center
PO Box 90319
DURHAM NC 27708-0319

Tel: 919 660-2695
Fax: 919 660-2671
Email: p.ingram-at-cellbio.duke.edu

==============================Original Headers==============================
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From: nizets2-at-yahoo.com
Date: Fri, 7 May 2010 01:28:17 -0500
Subject: [Microscopy] UV/Low Temp Resin and Other Embedding Questions

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

Hi Charlene,

I am no LR white specialist, but I remember some past discussions about it.
First of all you must make sure to avoid contact with oxygen in the air to allow polymerization.
Second, your must make sure that UV light can penetrate your sample (sorry if it seems stupid but sometimes the reasons are really stupid).
I hope that someone else can provide more specific help.

As for SiO2 sectionning, I wouldn't consider using a diamond knife.

As for the sulfur embedding, I am curious about it.
May I ask you to share with us the informations you got offline about it?

Regards,

Stephane



----- Original Message ----
X-from: "c-wilke-at-northwestern.edu" {c-wilke-at-northwestern.edu}
To: nizets2-at-yahoo.com
Sent: Fri, May 7, 2010 12:05:16 AM

Hello Listers,

I am trying to polymerize LR White at -20 C (no success after 24 
hours), in a Leica AFS unit. The UV system consists of 10 UV LEDs that 
are ~10 cm above the floor of the chamber. I can get the resin 
polymerizes in a 60 C oven in ~12 hours. If the problem is the LR 
White, can anyone advise me as to which resins work well with low 
temp., UV polymerization?

Other embedding questions;  I have a client that is embedding an 
agarose enrobed sample in silica. Can SiO2 be sectioned? Or would one 
prepare (polish) it for use in a SEM?
Another client would like to section soil by embedding in sulfur.
X-from what I understand, 1) the use of a cryo microtome is one method 
to acquire sections.
2) and a room temperature sulfur embedding process mentions "the 
plastic consistancy of the sulfur block, which is limited to 6-10 
hours, before it returns to an orthorhombic state", which is not 
compatible to sectioning.
Are these extremely complex processes? How does one work with sulfur? 
What kinds of safety and waste disposal precautions should I be aware 
of?

Thank you,
Charlene


Charlene Wilke
Imaging Specialist
Biological Imaging Facility
Northwestern University
2205 Tech Drive
Evanston, Illinois 60208
847.491.2842





==============================Original Headers==============================
10, 16 -- From c-wilke-at-northwestern.edu Thu May  6 17:01:28 2010
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28, 30 -- From: Stephane Nizet {nizets2-at-yahoo.com}
28, 30 -- Subject: Re: [Microscopy] UV/Low Temp Resin and Other Embedding Questions
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From: nizets2-at-yahoo.com
Date: Fri, 7 May 2010 01:38:41 -0500
Subject: [Microscopy] viaWWW: Zebra Fish Yolk

Contents Retrieved from Microscopy Listserver Archives
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I didn't search much more than 0,24 seconds but I found this protocol which may be worth a look:
http://cshprotocols.cshlp.org/cgi/content/abstract/2007/12/pdb.prot4772

Sorry no experience with zebrafish yolk here BUT
Given that zebrafishes and fixatives didn't change much in 50 years, protocols which worked well in the 60' may still work well today!
 
Good luck,

Stephane (without-an-i)



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Email: rra-at-stowers.org
Name: Rhonda Trimble

Organization: Stowers Institute

Title-Subject: [Filtered] Zebra Fish Yolk

Message: Has anyone out there ever successfully fixed and sectioned
through Zebra Fish Embryo's with the yolk?  My researcher wants to
see what is going on in the yolk.  I have no experience in the
matter.  I am going to do some HPF, but am also looking for a
chemical fixed method newer than the 1960's.

Thanks in advance!
Rhonda Trimble

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From reijma-at-luukku.com Fri May 7 05:28:32 2010





From: Frank_Karl-at-lincolnelectric.com
Date: Fri, 7 May 2010 06:08:50 -0500
Subject: [Microscopy] Re: UV/Low Temp Resin and Other Embedding Questions

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Hi Charlene,
I can't add much to this discussion, but I suspect you will need to quench
the sulfur from a molten state. Sulfur has several xtal states, and it does
form a unstable glassy state that has been described as "flexible". To my
knowledge S is not a hazardous waste and I suspect you can trash can it.
At least we could several years ago. It is flammable, so work in the hood,
watch your temperature and I suspect the embedding material needs to be
free of moisture, organics and oxidizers. Sulfur is soluble to some
degree in many organic solvents including isopropanol, chloroform and
toluene so be careful what you use to lubricate your blade. But you can
recover your media and sample if you need to.

In all the years I worked at a rubber company, we never thought to embed
anything in sulfur for thin-sectioning. Keep us informed!



c-wilke-at-northwest
ern.edu
To
05/06/2010 06:08 frank_karl-at-lincolnelectric.com
PM cc

Subject
Please respond to [Microscopy] UV/Low Temp Resin and
c-wilke-at-northwest Other Embedding Questions
ern.edu












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Hello Listers,

I am trying to polymerize LR White at -20 C (no success after 24
hours), in a Leica AFS unit. The UV system consists of 10 UV LEDs that
are ~10 cm above the floor of the chamber. I can get the resin
polymerizes in a 60 C oven in ~12 hours. If the problem is the LR
White, can anyone advise me as to which resins work well with low
temp., UV polymerization?

Other embedding questions; I have a client that is embedding an
agarose enrobed sample in silica. Can SiO2 be sectioned? Or would one
prepare (polish) it for use in a SEM?
Another client would like to section soil by embedding in sulfur.
From what I understand, 1) the use of a cryo microtome is one method
to acquire sections.
2) and a room temperature sulfur embedding process mentions "the
plastic consistancy of the sulfur block, which is limited to 6-10
hours, before it returns to an orthorhombic state", which is not
compatible to sectioning.
Are these extremely complex processes? How does one work with sulfur?
What kinds of safety and waste disposal precautions should I be aware
of?

Thank you,
Charlene


Charlene Wilke
Imaging Specialist
Biological Imaging Facility
Northwestern University
2205 Tech Drive
Evanston, Illinois 60208
847.491.2842





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From: TindallR-at-missouri.edu
Date: Fri, 7 May 2010 15:01:38 -0500
Subject: [Microscopy] EM Techs salaries

Contents Retrieved from Microscopy Listserver Archives
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Dear Collective,

Any feeling for what a recently graduated MACT or Delta electron microscopy technician might be pulling down as an entry-level salary in a university setting these days?

Just curious.....

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




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From: Vk69-at-Cornell.edu
Date: Fri, 7 May 2010 18:34:32 -0500
Subject: [Microscopy] viaWWW: Buying diamond knife for mcrotoming polymer nanofibers

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Email: Vk69-at-Cornell.edu
Name: Vibha

Organization: Cornell univ

Title-Subject: [Filtered] Buying diamond knife for mcrotoming polymer
nanofibers

Message: I amlooking to buy a diamond knife from electron microscoy
sciences for sectioning polymeric nanofibers. I mostly do room temp.
Microtoming and may do cryogenic at times. I have used a room temp
knife from electron microscopy sciences (35 deg) in the past and it
works very well foR me. Now I am thinking if I should get the
cryogenic wet knife (35 deg) so I can use it for both room temp and
cryogenic. Does anyone know how well does the cryogenic knife section
at room temp compared to the rom temp knife?

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From: kraftpiano-at-gmail.com
Date: Fri, 7 May 2010 23:14:45 -0500
Subject: [Microscopy] Pfeiffer Turbo Pump question.

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

I have a Pfeiffer TPH-270 and a TCP-300 with the connection cable that
joins them. The controller does not have relay K7 in place. I have
the cable that goes to the mains connection, but the cable has been
cut. When I look at the documentation, it tells me to put power to
pins a2 and b1 (a2 being neutral, B1 being the live wire, and A6 being
ground.) There is nothing indicating that the controller unit is
getting any power at all. I've tried everything that I can
comfortably do without further documentation, including checking all
of the fuses, and tracing out the connections inside the controller to
see if there was a fuse I missed somehow.

I can't get the pump to spin up, nor can I even get the unit to power
on. (Not that there's a power switch, but I'd expect I'd see
something- a backlight on the rotation meter perhaps?)

Does anyone have experience with these pumps and can give me a little
guidance on how to get it working? The documentation is really
sketchy. And mostly in German. I only speak English and French.

Thanks,

Justin A. Kraft

--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar

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From: edelmare-at-muohio.edu
Date: Sat, 8 May 2010 08:15:05 -0500
Subject: [Microscopy] Re:Nano Particles for SEM

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


I have treated nano particles for SEM like those for TEM. Simply
place a drop of solution on a carbon coated substraited TEM Grid,
allow to settle, wick off excess solution (from the edge). "Mount"
the grid on an SEM stub with two slightly angled (5-10 degrees)
strips of carbon tape spaced approximately 3mm apart. Stick the grid
on to the tape strips at the point where the grid is just slightly
touching the tape (makes for easy removal). These samples can be
used for both SEM and TEM imaging.



On 22 Apr 2010 at 22:40, fahayes-at-ucdavis.edu wrote:

}
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} Can someone suggest a sample prep method for mounting and/or coating
} 50nm-150nm sized Zinc Oxide particles suspended in water for SEM
}
} Thanks in advance
}
} Fred Hayes
} Erica McKenzie
} UC Davis
}
}
} ==============================Original
} Headers============================== 4, 25 -- From
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Richard E. Edelmann, Ph.D.
EXPO Editor, Microscopy and Microanalysis Supplement
Electron Microscopy Facility Director
364 Pearson Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-muohio.edu
http://www.emf.muohio.edu


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From: allan.mitchell-at-stonebow.otago.ac.nz
Date: Sun, 9 May 2010 16:59:31 -0500
Subject: [Microscopy] Lowicryl HM20 polymerisation problem.

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

Hi All

I have just removed a 'batch' of brain slices embedded in Lowicryl
HM20 resin from our Leica AFS freeze substitution instrument. There
are 24 blocks in total. Most are yellow tinge colour, and 8 are
tinged purple. Of the purple tinged blocks 5 are tinged a light
purple throughout while the remaining 3 are only tinged purple at the
top of the block (opposite end to the tissue sample). The yellow
tinged blocks are evenly tinged. Normally we get transparent, clear
blocks.

Our UV polymerisation schedule in the AFS is;

24 hours at -50oC
Rises to -10oC over four hours then remains at -10oC for a further 20
hours.
Rises to +15oC over 2.5 hours then remains at + 15 for 21.5 hours

Total polymerisation time of 96 hours.

While polymerising the resin (in Riechert special embedding capsules
that are inserted into gelatine capsules and mounted on the spider
holder - the normal method for the AFS ) the blocks are suspended in
an ethanol heat sink. This is normal AFS procedure.

The only difference in this run to all other runs with brain slices we
have done is that we incorporated 0.5% uranyl acetate in the methanol
cryosubstitution fluid. The reason for this is we normally work with
cerebellum and hippocampus, but has started on thalamus and are
getting lousy myelin preservation in the very myelin rich areas of the
thalamus.

Has anybody come across this purple tinge, or yellow tingle with their
polymerised Lowicryl HM20 blocks before? Any thoughts on what may be
going on?

I have not attempted sectioning the blocks yet so have no comment to
make yet about sectioning characteristics, or labeling
characteristics. That may come later in the week.

Thanks

Allan

.


Allan Mitchell
Microscopy Otago - Electron Microscopy
c/- Department of Anatomy and Structural Biology
Otago School of Medical Sciences
P.O. Box 913
Dunedin
New Zealand

Phone (03) 479 5642 or 479 7301
Fax (03) 479 5086 or 479 7254

EM Centre: http://ocem.otago.ac.nz/
Confocal Centre: http://occm.otago.ac.nz/
NZ Microscopy Society: http://microscopynz.co.nz/


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From: dxb430-at-googlemail.com
Date: Mon, 10 May 2010 07:43:31 -0500
Subject: [Microscopy] viaWWW: SEM gas flow drying effect

Contents Retrieved from Microscopy Listserver Archives
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Email: dxb430-at-googlemail.com
Name: Danny Bayliss

Organization: Loughborough University

Title-Subject: [Filtered] SEM gas flow drying effect

Message: Hi All,

I have been looking at bacteria that are filter deposited on to
membrane filters to see how they are distributed across the membrane
and I have been obtaining some good even deposits across the membrane.

I am conducting treatments to these bacteria after they are filter
deposited which involves a gas flow that will directly impact the
bacteria. If I fix and prepare my mambrane samples for SEM after a
gas flow has been applied to the membrane I observe a ring patterning
of the bacteria across the membrane with no damage to the bacterial
structure.

I was just wondering whether anyone has observed such a pattern or
know of published work that show this patterning? All I seem to find
in the literature are drying effects from sample preparations.

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From: RossLM-at-missouri.edu
Date: Mon, 10 May 2010 13:27:20 -0500
Subject: [Microscopy] 8th Annual Image Processing and Analysis Workshop, June 8-11

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

If I understand what you are describing, it sounds like the bacteria dried out along the edges of a bubble or droplet of water.

Thomas E. Phillips, Ph.D.
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
Biological Sciences
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (voice)
573-882-0123 (fax)


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To: Phillips, Thomas E.

This Question/Comment was submitted to the Microscopy Listserver
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Email: dxb430-at-googlemail.com
Name: Danny Bayliss

Organization: Loughborough University

Title-Subject: [Filtered] SEM gas flow drying effect

Message: Hi All,

I have been looking at bacteria that are filter deposited on to
membrane filters to see how they are distributed across the membrane
and I have been obtaining some good even deposits across the membrane.

I am conducting treatments to these bacteria after they are filter
deposited which involves a gas flow that will directly impact the
bacteria. If I fix and prepare my mambrane samples for SEM after a
gas flow has been applied to the membrane I observe a ring patterning
of the bacteria across the membrane with no damage to the bacterial
structure.

I was just wondering whether anyone has observed such a pattern or
know of published work that show this patterning? All I seem to find
in the literature are drying effects from sample preparations.

Login Host: 131.231.202.31
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From tnsct7779-at-india.com Mon May 10 09:16:21 2010
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The Electron Microscopy Core at the University of Missouri is hosting the 8th Annual Short Course and Workshop on Computer-Assisted Image Analysis and Measurement taught by Dr. John C. Russ on June 8-11, 2010. This popular course is intended to familiarize users of image analysis hardware and software with the fundamental principles and methods available to obtain meaningful results.

Image analysis and measurement techniques are utilized in a broad range of applications usually to extract numerical values (number, size, shape, etc.) or the location of objects. In other cases, global structural parameters such as the volume and surface of features are of interest. These measurements may require image processing to correct defects, enhance features, compare multiple images, recognize objects, or other steps. Measurements on individual features, or on the image as a whole, must then be obtained and interpreted in a proper stereological context to obtain useful data. Statistical interpretation of the data allows comparisons of different populations, understanding of distribution plots, and other inferences about the original objects. Structural modeling and geometric probability can be used to develop models for this interpretation.

The course relies heavily on tightly coupled lectures and hands-on exercises covering a wide variety of methodologies, approaches and tools, through a set of practical, step-by-step instructions to minimize the learning curve. Time will also be allotted for students to develop imaging strategies while working on their own micrographs.

No specific background is assumed, although users should already be familiar with microscopy or other imaging technologies. The software platform for the examples and exercises is Adobe Photoshop utilizing a comprehensive set of plug-ins from Reindeer Graphics. The course begins with An Introduction to Adobe Photoshop for Scientific Imaging.

Dr. Russ is the internationally acclaimed author of innumerable articles and several books on image analysis techniques and digital imaging, including the The Image Processing Handbook. He is widely known for his workshops and short courses and has helped to develop software packages to make image analysis methods more accessible to non-specialists. Many of the examples used in the course involve light or electron microscope images, but students are invited to bring their own images (TIFF files) for discussion and analysis. Ample time is provided at the end of the course for individual instruction.

The registration fee is only $1400. Enrollment is limited and there are still a few seats available. More information can be found at: www.emc.missouri.edu/russcourse or by contacting the course coordinator Lou Ross.

Lou Ross
Sr. Research Specialist
University of Missouri
Electron Microscopy Core Facility
W136 Veterinary Medicine Building
Columbia, MO 65211
573.882.4777, fax 573.884.2227
RossLM-at-missouri.edu
http://www.emc.missouri.edu/



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From: DDOE-at-wsc.ma.edu
Date: Mon, 10 May 2010 20:10:07 -0500
Subject: [Microscopy] TEM Zeis EWM 9S2 roughing pump

Contents Retrieved from Microscopy Listserver Archives
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Doug,
I had a similar problem last week with some nerve tissue. It was
embedded in EMbed-812 (Epon alternative). However, the tissue was
brittle and crumbled only in some areas and I am quite certain that it
is related to poor fixation in those areas. I suggest that you
participate in the fixation at least once and make sure that the tissues
blocks are minced quickly and with minimal mechanical distortion to a
size about 1mm X 1mm. The prolonged storage is not helpful but may be
unavoidable. Let us know if you find the solution.

Larry

drk-at-shcc.org wrote:
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} Hello All,
}
} Thank you for those many that have tried to help with my problems with fixation and embedding of liver samples. There have been many good suggestions but so far none solves my problem.
}
} I would like to be a bit more specific in describing our protocol just in case someone might think of another possible cause.
}
} To recap, we have had consistent problems with extremely brittle blocks when cutting thick or ultrathin sections of liver. The tissue (not the surrounding blank epon) fractures like glass, even during trimming with a double edge razor blade. We use the same protocol we use for the many other tissues that we embed well, so this problem is specific to liver. Our protocol includes primary phosphate buffered fixative (1.5% glut/1.5% parform with 0.05% tannic acid) followed by 1% OsO4 (with appropriate buffer rinses), dehydration in EtOH to 100%, a rinse in PO and infiltration and embedding in Spurrs resin, mixed according to Ellis. We have also embedded in EPON 812 substitute with similar brittleness. As I say, we use this protocol successfully for all other tissues.
}
} One suggestion received is that there is some water left in the sample which makes embedded tissue in Spurrs very brittle. I am sure our ethanol, PO and media is dry, but is it necessary to dehydrate liver longer than other tissues?
}
} The tissues are initially fixed in another laboratory, then sent to us. The tissue pieces are about 2mm x 2mm but were fixed in two changes of primary fixative for several hours each; then stored for a couple of weeks, then sent to us. Once in my laboratory, they are cut down so that one face is no larger than 0.5mm. Then they are osmicated, dehydrated and embedded. Could either the initial excessive size of the tissues, excessive fixation time or storage in buffer be a factor?
}
} Does anyone have another suggestion? If you work successfully with liver, would you mind sending a protocol?
}
} Many thanks,
}
} Doug
}
} Douglas R. Keene
} Assistant Investigator
} Micro-Imaging Center
} Shriners Hospital for Children
} 3101 S.W. Sam Jackson Park Road
} Portland, Oregon 97239
} drk-at-shcc.org
}
}
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--
Larry Ackerman, Specialist
Electron Microscopy Lab
UCSF, Dept. of Anatomy, Rm S1355
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

415-999-4758


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From howard.lee-at-guoco.com Mon May 10 14:19:05 2010
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I just went to switch out the 50W/AC L1 bulb on my Zeiss Axiophot's fluores= cence lamp housing. The new bulb looks to be a totally different design. It has a metal disk on one side of the bulb and there are plates above and below the bulb portion in the center. Did the design of these bulbs get changed or do I have the wrong ones? I buy a big batch of bulbs at one time so I don't know when I bought these. Would the disk on the center bulb region go to the right or left orientation (i.e., not front or back)? Thanks for any insight you might have. Tom


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/




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From tmartin4-at-travelers.com Mon May 10 18:12:28 2010
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Is anyone familiar with the operation and repair of the roughing pump for a Zeiss electron microscope model EM 9S2, vintage early 1970's. Ours pumps for awhile after turning the scope on , but then shuts off before the vacuum reaches a level for the diffusion pump to be turned on. We're not sure if the problem is in the roughing pump or perhaps the diffusion pump, but our service tech thinks it's the roughing pump. The pump came with the scope and was made in Germany. The specifications below are stamped on the pump motor and one of the 2 vacuum chambers. Please contact me directly if you can be of assistance. Thank you.

Vacuum pump information

Motor-
Electronmotoren by Werke Kaiser
Model # 1147077
Type DN 71 A/2 J-KL E
220/380 V 1.73A
.5 psi .37 kw 60 hz

There are 2 vacuum chambers
Type RZ-12
Sauvgermogen (vacuum rate- I think) – 12M3/h
Endpartialdruck (lowest pressure I think) – 6X10-5 Torr


Dr. David A. Doe
Biology Department
Westfield State College
577 Western Ave
Westfield, MA 01086
413-572-5291
fax: 413-572-8109


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From: DDOE-at-wsc.ma.edu
Date: Mon, 10 May 2010 20:27:42 -0500
Subject: [Microscopy] correction to subject title - TEM Zeis EM 9S2 roughing pump

Contents Retrieved from Microscopy Listserver Archives
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There was a typo in the subject title of my previous message. The TEM is a Zeiss EM 9S2.

Dr. David A. Doe
Biology Department
Westfield State College
577 Western Ave
Westfield, MA 01086
413-572-5291
fax: 413-572-8109

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From: stefan.diller-at-t-online.de
Date: Tue, 11 May 2010 02:01:40 -0500
Subject: [Microscopy] Re: TEM Zeis EWM 9S2 roughing pump

Contents Retrieved from Microscopy Listserver Archives
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Hello David,
I did a lot of repair on EM 9 because I still have two running at highschools here in Germany :-)

1- To find out what might be wrong, you first need to be sure that the roughing pump goes below 0,1 Torr at least in pressure when
you measure it unhooked from the vacuum system of the EM9.
You wrote that the pump switches itself off after some time. That might indicate overheating the pump motor (try to change the
roughing pump oil with a less viscous one...), because there is no security switch in the EM9 itself to switch off the pump as you
described.
2- If the end pressure of the pump will go below 0,1 Torr you are on the working pressure needs of the diffusion pump.
3- Connect the roughing pump back to the EM9.
4- If your EM9 does have an automatic pump cycle controller (right hand at the rear on the console, take off cover) you can use
the first two plugs (black and red) to measure the vacuum. Atmosphere pressure is 10 volts, switch point pressure for going to
high vac is 1,7 volts, high tension on should be at 1,5 volts or below.
5- You can manually go through the pump cycle by interchanging those large black connectors, one plugged in at the cycle
controller, the other at the EM9 (before doing this, switch off EM9...). Then you can manually rough pump the column by pressing
"1" at the front panel and after 2 minutes finally go to "2" to high vac mode. You should be able to measure the pressure to your
roughing pump doing this. It should go down to at least 0,2 to 0,1 Torr at "1" and "2".
6- If roughing pressure will not go to 0,1 Torr and the roughing pump itself will, you have a leaking magnetic valve (at the left
side of the high vac pump setup. That might be a big problem because it is difficult to get the appropriate o-ring for repair.

If you need any more help or a complete set of electrical and technical plans of the EM9 as PDFs, please contact me offline.

Hope, this helps,
Stefan


--
-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
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-----------------------------------------------------



Am 11.05.10 03:16, schrieb DDOE-at-wsc.ma.edu:
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} Is anyone familiar with the operation and repair of the roughing pump for a Zeiss electron microscope model EM 9S2, vintage early 1970's. Ours pumps for awhile after turning the scope on , but then shuts off before the vacuum reaches a level for the diffusion pump to be turned on. We're not sure if the problem is in the roughing pump or perhaps the diffusion pump, but our service tech thinks it's the roughing pump. The pump came with the scope and was made in Germany. The specifications below are stamped on the pump motor and one of the 2 vacuum chambers. Please contact me directly if you can be of assistance. Thank you.
}
} Vacuum pump information
}
} Motor-
} Electronmotoren by Werke Kaiser
} Model # 1147077
} Type DN 71 A/2 J-KL E
} 220/380 V 1.73A
} .5 psi .37 kw 60 hz
}
} There are 2 vacuum chambers
} Type RZ-12
} Sauvgermogen (vacuum rate- I think) – 12M3/h
} Endpartialdruck (lowest pressure I think) – 6X10-5 Torr
}
}
} Dr. David A. Doe
} Biology Department
} Westfield State College
} 577 Western Ave
} Westfield, MA 01086
} 413-572-5291
} fax: 413-572-8109
}
}
} ==============================Original Headers==============================
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From: TindallR-at-missouri.edu
Date: Tue, 11 May 2010 08:57:20 -0500
Subject: [Microscopy] TEM: collagen fiber size and fixative

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

Hello!
I perform force spectroscopy experiments with AFM and I am trying to
estimate Young's modulus of a sample from a force curve (force vs.
distance). Although there is a huge pile of articles on that topic, I
am currently looking for special software which could reliably
calculate the Young's modulus from AFM data. I don't use SPIP because
it is not freeware. I am currently trying to use IgorPro (I have 4.08
version), but this program seems really complicated.
Could anybody help me with IgorPro or advice another program to
calculate Young's modulus?
--
Sincerely yours,
Dmitry Bagrov

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From c-jason.orial-at-mci.com Tue May 11 05:07:53 2010
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Dear Collective,

A client is looking at tissue from control vs. treated animals and measuring collagen fiber size, and he has observed that the collagen fibers in the area of interest in the tissue is smaller in the controls than in the treated samples. The problem is that it turns out the control samples were collected into formalin (because there was no EM fixative directly at hand at that time) and the treated tissues were collected into EM fix (2% glut/2% paraform in 0.1M cacodylate buffer).

To my antiquated way of thinking, this invalidates a direct comparison of the two tissues, but we are still curious as to whether one would expect the size difference to be attributable to the difference in primary fixations. Does anyone know off the top of their heads if this might be the case? (I'm admittedly being lazy here----busy day and not a lot of time to search around for tissue shrinkage rates in various fixatives.)

BTW, he claims that in one portion of the tissue the fiber sizes are the same, but in the area of interest they are different. Nothing's ever easy.....

Thanks in advance.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: stefan.diller-at-t-online.de
Date: Tue, 11 May 2010 13:09:17 -0500
Subject: [Microscopy] TEM Zeis EWM 9S2 roughing pump

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

Formalin is often stabilized with an alcohol such as 8% methanol. This would clearly affect shrinkage. But see also:
"Optimizing glutaraldehyde crosslinking of collagen: effects of time, temperature and concentration as measured by shrinkage temperature" JM Ruijgrok, JR De Wijn, ME Boon (1994) Journal of Materials Science: Materials in Medicine
5(2):1573-4838.




Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Tuesday, May 11, 2010 8:58 AM
To: Phillips, Thomas E.

Dear Collective,

A client is looking at tissue from control vs. treated animals and measuring collagen fiber size, and he has observed that the collagen fibers in the area of interest in the tissue is smaller in the controls than in the treated samples. The problem is that it turns out the control samples were collected into formalin (because there was no EM fixative directly at hand at that time) and the treated tissues were collected into EM fix (2% glut/2% paraform in 0.1M cacodylate buffer).

To my antiquated way of thinking, this invalidates a direct comparison of the two tissues, but we are still curious as to whether one would expect the size difference to be attributable to the difference in primary fixations. Does anyone know off the top of their heads if this might be the case? (I'm admittedly being lazy here----busy day and not a lot of time to search around for tissue shrinkage rates in various fixatives.)

BTW, he claims that in one portion of the tissue the fiber sizes are the same, but in the area of interest they are different. Nothing's ever easy.....

Thanks in advance.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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21, 34 -- From PhillipsT-at-missouri.edu Tue May 11 10:28:42 2010
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From hnewell-at-usesgroup.com Tue May 11 10:46:30 2010
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...
Attention, I forgot something important in my first email:

When you are taking off the roughing pump vacuum connection to the EM9 but then switch on the pump via the EM9 paneel, you first
have to make sure that the magnetic valve on the left hand side of the high vac pumping system will not switch on after 2-3
minutes delay.
You have to take out the fuse "Si2" on the first electronic paneel on the right hand side. This will cut the voltage for the
magnetic valve.
Otherwise you might risk to fry the coil in the magnetic valve if you are working the valve against atmospheric pressure...

Best wishes,
Stefan



Am 11.05.10 09:06, schrieb stefan.diller-at-t-online.de:
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}
} Hello David,
} I did a lot of repair on EM 9 because I still have two running at highschools here in Germany :-)
}
} 1- To find out what might be wrong, you first need to be sure that the roughing pump goes below 0,1 Torr at least in pressure when
} you measure it unhooked from the vacuum system of the EM9.
} You wrote that the pump switches itself off after some time. That might indicate overheating the pump motor (try to change the
} roughing pump oil with a less viscous one...), because there is no security switch in the EM9 itself to switch off the pump as you
} described.
} 2- If the end pressure of the pump will go below 0,1 Torr you are on the working pressure needs of the diffusion pump.
} 3- Connect the roughing pump back to the EM9.
} 4- If your EM9 does have an automatic pump cycle controller (right hand at the rear on the console, take off cover) you can use
} the first two plugs (black and red) to measure the vacuum. Atmosphere pressure is 10 volts, switch point pressure for going to
} high vac is 1,7 volts, high tension on should be at 1,5 volts or below.
} 5- You can manually go through the pump cycle by interchanging those large black connectors, one plugged in at the cycle
} controller, the other at the EM9 (before doing this, switch off EM9...). Then you can manually rough pump the column by pressing
} "1" at the front panel and after 2 minutes finally go to "2" to high vac mode. You should be able to measure the pressure to your
} roughing pump doing this. It should go down to at least 0,2 to 0,1 Torr at "1" and "2".
} 6- If roughing pressure will not go to 0,1 Torr and the roughing pump itself will, you have a leaking magnetic valve (at the left
} side of the high vac pump setup. That might be a big problem because it is difficult to get the appropriate o-ring for repair.
}
} If you need any more help or a complete set of electrical and technical plans of the EM9 as PDFs, please contact me offline.
}
} Hope, this helps,
} Stefan
}
}

--
-----------------------------------------------------
Stefan Diller - Wissenschaftliche Photographie
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

==============================Original Headers==============================
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From: R.J.Fairhurst-at-exeter.ac.uk
Date: Wed, 12 May 2010 09:47:29 -0500
Subject: [Microscopy] willemite and hemimorphite

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

...
Attention, I forgot something important in my first email:

When you are taking off the roughing pump vacuum connection to the EM9 but then switch on the pump via the EM9 paneel, you first
have to make sure that the magnetic valve on the left hand side of the high vac pumping system will not switch on after 2-3
minutes delay.
You have to take out the fuse "Si2" on the first electronic paneel on the right hand side. This will cut the voltage for the
magnetic valve.
Otherwise you might risk to fry the coil in the magnetic valve if you are working the valve against atmospheric pressure...

Best wishes,
Stefan






Am 11.05.10 09:06, schrieb stefan.diller-at-t-online.de:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} Hello David,
} I did a lot of repair on EM 9 because I still have two running at highschools here in Germany :-)
}
} 1- To find out what might be wrong, you first need to be sure that the roughing pump goes below 0,1 Torr at least in pressure when
} you measure it unhooked from the vacuum system of the EM9.
} You wrote that the pump switches itself off after some time. That might indicate overheating the pump motor (try to change the
} roughing pump oil with a less viscous one...), because there is no security switch in the EM9 itself to switch off the pump as you
} described.
} 2- If the end pressure of the pump will go below 0,1 Torr you are on the working pressure needs of the diffusion pump.
} 3- Connect the roughing pump back to the EM9.
} 4- If your EM9 does have an automatic pump cycle controller (right hand at the rear on the console, take off cover) you can use
} the first two plugs (black and red) to measure the vacuum. Atmosphere pressure is 10 volts, switch point pressure for going to
} high vac is 1,7 volts, high tension on should be at 1,5 volts or below.
} 5- You can manually go through the pump cycle by interchanging those large black connectors, one plugged in at the cycle
} controller, the other at the EM9 (before doing this, switch off EM9...). Then you can manually rough pump the column by pressing
} "1" at the front panel and after 2 minutes finally go to "2" to high vac mode. You should be able to measure the pressure to your
} roughing pump doing this. It should go down to at least 0,2 to 0,1 Torr at "1" and "2".
} 6- If roughing pressure will not go to 0,1 Torr and the roughing pump itself will, you have a leaking magnetic valve (at the left
} side of the high vac pump setup. That might be a big problem because it is difficult to get the appropriate o-ring for repair.
}
} If you need any more help or a complete set of electrical and technical plans of the EM9 as PDFs, please contact me offline.
}
} Hope, this helps,
} Stefan
}
}

--
-----------------------------------------------------
Stefan Diller - Wissenschaftliche Photographie
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

==============================Original Headers==============================
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From john.deland-at-cgi.com Tue May 11 14:26:17 2010
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Email: dainis-at-dauksta.com
Name: Dainis Dauksta

Organization: City and Guilds Art School London

Title-Subject: [Filtered] cheap microscope cameras

Message: Dear All

I'm thinking of purchasing a cheap digital microscope camera for the
school. Has anyone had any experience of the Chinese or Indian made
cameras; any recommendations?

Also does anyone have any ideas to get the quickest results in the
preparation of wood samples for microtomy please?

Regards

Dainis


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From brewstern-at-stginc.com Wed May 12 03:32:30 2010
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Dear colleagues,
Does anyone have experience with the luminix lxsr-scinti? Is it
worth the purchase if the P47 must be changed now?

Regards
Gerd

--
Gerd Mueller von der Haegen
BIOLAB Umweltanalysen GmbH
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tel: +49-(0)531-313000
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www.biolab.de




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I would also be interested in any information on the analysis of Hemimorphite and Willemite. I have recently analysed Hemimorphite (confirmed via optical microscopy) via EPMA and obtained the same results. It was suggested that the results are due to the water being driven off by interaction with the electron beam, is this possible?

Regards
Rob

-----Original Message-----
X-from: [mailto:renaudgeological-at-execulink.com]
Sent: 30 April 2010 23:46
To: Fairhurst, Robert

Hello All,

The reason I am writing is that I am currently working on a project
involving willemite and hemimorphite and would like to ask a question. The
rocks I am working on supposedly only contain hemimorphite, however, I have
come across what I believe to be "willemite". I analyzed the hemimorphite
and achieved a total of ~92-93 wt% (66 wt% ZnO and 26 wt% SiO2) which allows
for 7-8wt% water. I analyzed the supposed "willemite" and I get totals of
~100 wt% (73 wt% ZnO and 27 wt% SiO2). As a confirmation, I submitted the
grain for micro-xrd and the structure of the "willemite" came back as the
hemimorphite structure. So it turns out I have several grains which mineral
chemically resemble "willemite" but structurally resemble hemimorphite.
Have you seen this before and can you recommend any papers which may relate
to this.

Thanks in advance.

Jim.


==============================Original Headers==============================
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From: hstahlberg-at-me.com
Date: Thu, 13 May 2010 08:40:06 -0500
Subject: [Microscopy] [2dx] email server for electron crystallography image processing

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

I have only recently begun using a probe and I have been impressed by the effects of the beam. I operate the SEM well under 1 nA and occasionally see changes due to the beam. I operate the probe at 50 times higher currents and it is quite common to see beam-induced effects in the mounting material.

I would think it entirely possible that you are driving off water. Have you tried the same measurement at lower currents (or bigger spot sizes) to see how the totals turn out?

Warren Straszheim
ISU MARL/Ames Lab
_______________________
X-from: R.J.Fairhurst-at-exeter.ac.uk [R.J.Fairhurst-at-exeter.ac.uk]
Sent: Wednesday, May 12, 2010 9:48 AM
To: wesaia-at-iastate.edu

I would also be interested in any information on the analysis of Hemimorphite and Willemite. I have recently analysed Hemimorphite (confirmed via optical microscopy) via EPMA and obtained the same results. It was suggested that the results are due to the water being driven off by interaction with the electron beam, is this possible?

Regards
Rob

-----Original Message-----
X-from: [mailto:renaudgeological-at-execulink.com]
Sent: 30 April 2010 23:46
To: Fairhurst, Robert

Hello All,

The reason I am writing is that I am currently working on a project
involving willemite and hemimorphite and would like to ask a question. The
rocks I am working on supposedly only contain hemimorphite, however, I have
come across what I believe to be "willemite". I analyzed the hemimorphite
and achieved a total of ~92-93 wt% (66 wt% ZnO and 26 wt% SiO2) which allows
for 7-8wt% water. I analyzed the supposed "willemite" and I get totals of
~100 wt% (73 wt% ZnO and 27 wt% SiO2). As a confirmation, I submitted the
grain for micro-xrd and the structure of the "willemite" came back as the
hemimorphite structure. So it turns out I have several grains which mineral
chemically resemble "willemite" but structurally resemble hemimorphite.
Have you seen this before and can you recommend any papers which may relate
to this.

Thanks in advance.

Jim.


==============================Original Headers==============================
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5, 22 -- From: "Renaud Geological Consulting Ltd." {renaudgeological-at-execulink.com}
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From kristin-at-kristinlong.com Wed May 12 13:05:40 2010
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Hello,
I've got a couple of Leitz microscopes that need service for stage focus and eyepiece motion. Does anyone have recommendations for service on the west coast US?

Thanks,
Glen


Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923 USA
(206) 616-4156
glenmac-at-uw.edu










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From hofbauer-at-atecaudio.com Wed May 12 19:43:33 2010
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{zaluzec-at-microscopy.com} ; Thu, 13 May 2010 02:43:30 +0200


Hi,

We would like to inform about a new [2dx] email list server that we
are hosting at the University of Basel. This email list is primarily
intended for users of the electron crystallography image processing
software 2dx, which is based on the MRC program suite that Richard
Henderson and others develop, and the MRC software itself, and the
IPLT image processing software system. If you are using one of these
software systems to process electron crystallography images of 2D
crystals of membrane proteins, or if you use one of these software
systems for the processing of crystal images in material sciences,
then we invite you to subscribe to this list.

This list should be a useful resource for discussing or posting
questions relating to electron crystallography sample preparation,
data collection, or image processing.

If you once downloaded the 2dx software, then you are one of the 385
people that are already subscribed to this list (it is easy to
unsubscribe).
If not, then we hereby invite you to subscribe to this list, by
accessing this web site:
https://www.maillist.unibas.ch/mailman/listinfo/2dx

We will closely monitor this email list, and promise that you will not
receive spam through this list.

All the best,

Henning

___________________________________________________

Henning Stahlberg
Center for Cellular Imaging and Nano Analytics (C-CINA)
at the Department of Biosystems Science and Engineering (D-BSSE)
Structural Biology and Biophysics, Biozentrum,
WRO-1058, Mattenstrasse 26
University Basel, CH-4058 Basel, Switzerland
Tel: +41-61-387 32 62 (office)
Tel: +41-61-387 32 31 (Karen Bergmann, administrative assistant)
Fax: +41-61-387 39 86
mailto:Henning.Stahlberg-at-unibas.ch
Skype:henningstahlberg
http://c-cina.org
http://2dx.org
___________________________________________________







==============================Original Headers==============================
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From: mcauliff-at-umdnj.edu
Date: Thu, 13 May 2010 12:25:31 -0500
Subject: [Microscopy] osmium + ferrocyanide and EmBed 812

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


Hi,

Two PhD and/or Postdoc positions are available immediately in the 2dx
software development team in our Center for Cellular Imaging and Nano
Analytics in the University of Basel, Switzerland.

The project involves the development of algorithms and implementation
of these for the 3D structure reconstruction of membrane proteins from
electron microscopy images. Several aspects of this task will be
addressed, including analysis and simulation of the image formation
process in the microscope, processing of the very noisy and enormously
large data sets involving maximum likelihood and other algorithmic
approaches, and the creation of user-friendly GUIs front end
interfaces, and parallelized (including GPU support) back-end
processing cores. This software will be integrated into the 2dx
software package that our group has produced, and which now enjoys
over 400 external users (http://2dx.org).
In addition, the candidate(s) would also be involved in structure
determination projects in the fields of single particle cryo-EM and
electron crystallography.
The position is available immediately, long-term funding is secured. C-
CINA is equipped with outstanding equipment. We offer a welcoming
working environment and attractive salaries in a multidisciplinary
group.

Experience in software development (C++) is required. Knowledge of
CUDA, Qt, Matlab, F77, or Electron Microscopy image formation, or
molecular biology basic knowledge is welcome, but can also be acquired
here in the lab.

Basel is a culturally rich and beautiful city at the border between
Switzerland, France and Germany.
The lab language is English.

If you are interested, or if you know of somebody who might, or to
obtain further information, please contact:

Henning Stahlberg

Center for Cellular Imaging and Nano Analytics (C-CINA)
at the Department of Biosystems Science and Engineering (D-BSSE)
Structural Biology and Biophysics, Biozentrum,
WRO-1058, Mattenstrasse 26
University Basel, CH-4058 Basel, Switzerland
Tel: +41-61-387 32 62 (Henning Stahlberg)
Tel: +41-61-387 32 31 (Karen Bergmann, administrative assistant)
mailto:Henning.Stahlberg-at-unibas.ch
http://c-cina.org
http://2dx.org

==============================Original Headers==============================
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9, 25 -- From: Henning Stahlberg {hstahlberg-at-me.com}
9, 25 -- To: Microscopy-at-microscopy.com
9, 25 -- Subject: PhD and PostDoc position in software development for cryo-EM of
9, 25 -- membrane proteins
9, 25 -- Date: Thu, 13 May 2010 15:41:05 +0200
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From cabanapet-at-mail2dallas.com Thu May 13 10:37:44 2010
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{zaluzec-at-microscopy.com} ; Thu, 13 May 2010 17:37:42 +0200

Greetings all:

I embedded routinely processed mouse brain in EmBed 812, no problems.
The following week I embedded similar tissue processed the same EXCEPT
that I used an osmium ferrocynide mix for osmication instead of plain
buffered osmium.
All other steps were identical.
The Os-ferrocyanide brain is impossible to cut, acts like it was not
infiltrated. Pure plastic cuts fine. Is it possible that Os-ferrocyanide
reacts with something in the EmBed 812??

Geoff

--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff-at-umdnj.edu
**********************************************



==============================Original Headers==============================
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From: CLJOHNSON-at-DREXEL.EDU
Date: Fri, 14 May 2010 08:58:30 -0500
Subject: [Microscopy] viaWWW: Image Analysis Software

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

I have done 1000+ blocks of osmium + ferrocyanide fixed tissues into epon-type resins without problem. I haven't used this combo in a while so I can remember whether I used it once the switch to epon 812 substitutes occurred but think I probably did. I doubt that is the problem.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: mcauliff-at-umdnj.edu [mailto:mcauliff-at-umdnj.edu]
Sent: Thursday, May 13, 2010 12:26 PM
To: Phillips, Thomas E.

Greetings all:

I embedded routinely processed mouse brain in EmBed 812, no problems.
The following week I embedded similar tissue processed the same EXCEPT
that I used an osmium ferrocynide mix for osmication instead of plain
buffered osmium.
All other steps were identical.
The Os-ferrocyanide brain is impossible to cut, acts like it was not
infiltrated. Pure plastic cuts fine. Is it possible that Os-ferrocyanide
reacts with something in the EmBed 812??

Geoff

--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
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Email: CLJOHNSON-at-DREXEL.EDU
Name: Craig Johnson

Organization: Drexel University

Title-Subject: [Filtered] Image Analysis Software

Message: Hello all,

I have some users who want information on image analysis software.
They are interested in something that can "automatically" extract
grain size, shape and distribution from SEM, FIB, and/or TEM images
of composite materials and metals. Any information the group might
have on capabilities and quality of available software packages will
be appreciated. One package they are looking at is SIMAGIS.

Thanks in advance,

Craig.


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From: tnkeim-at-sbcglobal.net
Date: Fri, 14 May 2010 08:58:30 -0500
Subject: [Microscopy] viaWWW: Question re Zeiss Epiplan objectives

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Email: tnkeim-at-sbcglobal.net
Name: Tom Keim

Organization: dentist

Title-Subject: [Filtered] Question re Zeiss Epiplan objectives

Message: My microscope uses (the old) 160mm tube length objectives. I
need transmitted light POL objectives with a flat field for use with
slides having standard coverslips. Problem: The Zeiss POL objectives
for transmitted light are not flat field, and the Epiplan POL
objectives are computed for uncovered specimens. Would someone who
has tried using the Epiplan POL objectives for transmitted light with
covered specimens please let me know how well that worked?

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From: paul_hazelton-at-umanitoba.ca
Date: Fri, 14 May 2010 12:02:28 -0500
Subject: [Microscopy] digital micrograph imaging software

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

25 years ago the idea of digital image capture was totally enamouring.
The ability to get rapid prints of clinical and diagnostic material was
fantastic. Even if the prints from the original fibre optic systems were
not to the same resolution, the improvement in Time to Report from
almost a week to less than a day was fantastic. And there was always wet
chemistry for publication prints or if we really needed resolution.

25 years later, as we finally get digital, I find the reality somewhat
less than captivating . But then, I have 40 years of wet chemistry
experience, and at the risk of sounding conceited, I am fairly good at
it. Of course, it could be argued that if you spend 40 years on it, you
should be able to teach a chimpanzee to make a relatively acceptable
print with wet chemistry. So hopefully this is not some closet Luddite
within who is battling his way out.


The problem is - we have had a used Gatan system installed, and are
capturing the images using Gatan DigitalMicrograph software. On the
monitor the images look alright, but just that - alright. Frankly, the
resolution is less than on the focusing screen, but for diagnostic, they
are alright. Hopefully I will be able to figure out how to actually
adjust the different brightness/contrast settings so that we can get
away from ‘averaged’ optimized data capture, which should improve the
original data.

The system falls apart when it is time to take the micrographs away to
process and print them. I have been using PhotoShop CS4 Enhanced, with
64 bit processing on my computer at home, and am not all that impressed.
Neither does Illustrator CS4 excite me. Ignoring the fine detail
resolution - there is no such thing as a fibre, forget it - the
immediate technical problem is getting acceptable prints. The adjustment
of contrast and brightness seems to be highly limited before bloom
effect takes over (Gee, using British/Canadian idiom, may I call it the
Blooming Effect!!!). There simply is not sufficient gray scale variance
to get a good micrograph. The contrast is too extreme, the background is
whited out due to saturation, etc.


There is going to be a need to upgrade the computer and printer hardware
and software. And I really want to do this right the first time. Which
leads to the questions

* What programs are people using that provide for better micrograph
processing,
* What printers are good. Frankly, the best we have here is not much
improved over my very old HP 1200 toner printer in the lab. Again,
as with processing the images, the best I can get is on my home
system, where I use an HP6400 multifunction system. And I don’t
like those prints.



So, thanks in advance. And for my fellow closet Luddites, don’t remind
me that it is, after all, a digital world, and the digital world is
[please insert your appropriate scatiological terminology representing
my favourite viral specimen source (fecal)]. We don't have to simply
accept the issues of pixel size, etc. We can do better.


Paul
--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 William Avenue
Winnipeg, Manitoba, Canada, R3E 0J9
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926


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From: mike.bode-at-resaltatech.com
Date: Fri, 14 May 2010 14:00:21 -0500
Subject: [Microscopy] digital micrograph imaging software (commercial response)

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

I'd like to respond to the recent posting by Paul Hazelton. However, I would
like to first put out a disclaimer:

As a person who has been involved with digital imaging on TEMS and SEMs for
a while, and who is currently selling these systems, I am definitely biased
towards digital imaging systems.

Now, Paul brings up a number of good points. Perhaps I can add some light on
this from another direction. There are many issues that have an effect on
the "quality" of images, and I think it's instructive to look at some of
them.

Let's start by looking at resolution, as it is often stated that "film has a
higher resolution". It turns out, in fact, that it is true. If you look at
the MTF of film compared to that of, for example. P47 (scintillator
material), which has been measured (references available, please ask), we
find that film has around twice the resolution (50% point of the MTF) than
the scintillator has. But the ultimate limit of the entire system is of
course determined by the resolution of the TEM. John Spence measured the MTF
of SO-163 and came up with about 32 lp/mm, or 30 micron, which also depends
on the development procedure. That means that if you have a TEM with 0.2 nm
resolution, you need a magnification of at least 150kx to see the detail on
film. For digital, you need to go to something like 300kx, but then you see
the same detail. You lose some field of view, but you can resolve whatever
the microscope can. So, resolution is clearly not the issue.

Well, then it must be the dynamic range. But that's not it either. Our eyes
have a dynamic range of maybe around 7 bits (we can distinguish perhaps 100
levels of grey). Film can have different dynamic range, depending on the
development process, but typically that would be 6-12 bit (64-4000 levels of
grey, the latter only with significant reduction in resolution). Digital
cameras can have resolutions between 12 and 16 bit, depending on various
factors such as cooling, etc. Now, everybody would agree that 64,000 levels
of grey are better than 200.

So why do some people see digital images as inferior? I think the answer
lies somewhere else. First of all, most people don't use the full dynamic
range of their cameras, and it may not always be possible. The information
may be "bunched" in a small range of grey levels, and then this information
is "stretched" and "shifted" to make it visible to the eye. In extreme
cases, one could wind up with only 50 levels of the 64,000 used, which would
then be visible in areas that have fine grey level details. So, look at the
histogram when you acquire images. If the data is all bunched up at one end
of the histogram, the images will probably not look good. For example, if
the background is all saturated, it is probably a good idea to reduce the
beam current or exposure time.

Then we have the linearity of the digital cameras.. As opposed to film, they
are very linear, which is a good thing, with a sharp cut-off at the low and
high intensities. Film, in contrast, has an S-shaped response curve (gamma).
This makes for pictures that are easier on the eyes, as the sharp cut-offs
at saturation are absent, but makes film much less useful as an analytical
tool. Of course, you can mimic that with a digital camera, as we can usually
set the display to various gamma values.

The last thing I think contributes is the fact that digital images are
usually observed on a monitor and are very easy to manipulate. We CAN go to
the extremes and beyond by simply clicking a button. We CAN magnify the
image on the screen to ridiculous proportions just by selecting it with the
mouse. That's not possible with film. You have to go back to the dark room
and play with chemicals for an hour. Because of that we have all learned
that we can't blow up a negative too much, and so we never see the effects.

Film definitely has an advantage in terms of information density. It can
record more information over a larger area. But digital has the advantage in
almost anything else, and the film's advantage can be overcome with things
like taking serial images and montaging them. It takes a bit longer, but
serves the same purpose.

Now for Paul's questions. I think that Photoshop and Illustrator are fine
products, but they are designed for color photos with the aim of publishing
them. Photoshop alone, for example, does not even have the concept of
magnification, unless you use some plug-ins. I am not so familiar with
Illustrator, but I guess it's the same story (this may be a good point to
remind everybody that I am earning money from selling a competing product
and you should come to your own conclusions). So, the first thing I would
suggest is to look at software that is made for microscopy, and go from
there. And you don't have to stick with what we are selling; there are many
fine products out there.

As far as the printer goes, it is a developing story. My suggestion would be
to look for a high quality ink jet printer and use high quality paper (photo
quality) for the best results. Get a good, but inexpensive laser printer for
the routine pictures. I have had good results with an Epson R800 Ink-jet
printer (I think they don't make them anymore). For further information send
me an email. I can give you a contact who is an expert in the field, but I
don't want to put out his name here and get him swamped with emails :-)
John, if you are listening, perhaps you could say a few words?

Mike Bode
---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode AT ResAltaTech.com
www.ResAltaTech.com






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From: PhillipsT-at-missouri.edu
Date: Mon, 17 May 2010 14:39:08 -0500
Subject: [Microscopy] LM Slide colors

Contents Retrieved from Microscopy Listserver Archives
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Hi Geoff

You raised an interesting topic. Last month, I posted an entry reporting some infiltration problems using EmBed 812. I use a similar protocol for Xenopus tadpole eyes (which are approximately 1mm3), involving osmicating the samples with potassium ferricyanide. As a result, the samples are (again) impossible to cut, although the ultrastructure seemed well preserved, indicating that there is a dehydration/ infiltration problem. I am currently modifying our standard protocol (basically increasing dehydration and infiltration times) and will consider removing the potassium ferricyanide, as I was already advised.

Hope this helps.

Tami



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

} Date: Thu May 13 10:37:17 PDT 2010
} From: mcauliff-at-umdnj.edu
} Subject: [Microscopy] osmium + ferrocyanide and EmBed 812
} To: tbogea-at-interchange.ubc.ca
}
}
}
}
} ----------------------------------------------------------------------------
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} Greetings all:
}
} I embedded routinely processed mouse brain in EmBed 812, no problems.
} The following week I embedded similar tissue processed the same EXCEPT
} that I used an osmium ferrocynide mix for osmication instead of plain
} buffered osmium.
} All other steps were identical.
} The Os-ferrocyanide brain is impossible to cut, acts like it was not
} infiltrated. Pure plastic cuts fine. Is it possible that Os-ferrocyanide
} reacts with something in the EmBed 812??
}
} Geoff
}
} --
} --
} **********************************************
} Geoff McAuliffe, Ph.D.
} Neuroscience and Cell Biology
} Robert Wood Johnson Medical School
} 675 Hoes Lane, Piscataway, NJ 08854
} voice: (732)-235-4583
} mcauliff-at-umdnj.edu
} **********************************************
}
}
}
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} ==============================End of - Headers==============================
--
Dr. Tami Bogea
Research Associate
Ophthalmology & Visual Sciences
University of British Columbia
2550 Willow St. Vancouver, BC
V5Z 3N9 Canada
Tel: 604-875-4648
Fax: 604-875-4663


==============================Original Headers==============================
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From cainfo-at-58degrees.com Fri May 14 14:18:39 2010
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Please note that Ms. Sanchez is asking about **all** types of
compound microscopes, not just the usual case.

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
==========================================================

} realname - Christine Sanchez
} Email - cvjurasinski-at-yahoo.com
} ORGANIZATION - Archdiocese of Philadelphia
} EDUCATION - 6-8th Grade Middle School
} SUBJECT_OF_QUESTION - Image formed in compound microscope
} QUESTION - I'm teaching a lesson using a compound microscope and
} several slides, including a letter "e" slide. The microscopes that
} we use in class produce an image of the letter "e" that is inverted
} and reversed (real image). I and my students are wondering if the
} images formed by ALL compound microscopes are always real. I
} thought I read somewhere that the image may differ depending upon
} whether a mirror, prism or other lenses (infinity systems) is used
} to direct the light between the objective and eyepiece. For
} example, if a prism is used, the letter "e" will appear reversed by
} not inverted.
}
} I'm having trouble finding an answer to this question on the
} internet. Much of what I find talks in generalities about the
} images formed in a compound microscope. Could you help to answer
} this question? Would I be wrong to tell the students that ALL
} compound microscopes produce a reversed, inverted (real) image of
} the specimen?
}
} Thank you,
}

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From reservas-at-danncarlton.com Fri May 14 14:57:31 2010
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Dear All,

I felt compelled to respond to your expression of anguish. I come from a background of photography, both fine art and scientific, and remember how a TEM final print from a point source enlarger could take your breath away. It's all digital now and I simply document everything at double the magnification as was used with film. This is a practical approach and gives all the information required to complete each project. However, for many years now, nothing has taken my breath away and I miss it. This isn’t just an old-timers lack of objectivity. When young people are in the lab and happen to come across an old transmission electron micrograph, it never fails to stop them in their tracks and take their breath away and I realize we really have lost something. The software issue, at times, can be the bit depth of the display being less than the bit depth of the image (if you have a full range image).
If you print the image on a printer capable of rendering the full greyscale it might look much better than what you can see on the computer screen. But it is regrettable that it requires you to guess at what may come out of a high end printer rather than having hands on contol and watch it come up into the soft glow of the safelight to take your breath away.

Robert Underwood
University of Washington/Dermatology







==============================Original Headers==============================
9, 24 -- From underwoo-at-u.washington.edu Fri May 14 16:12:14 2010
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From rhenriquez-at-cbselectpros.com Fri May 14 16:46:04 2010
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{microscopy-at-microscopy.com} ; Fri, 14 May 2010 16:45:56 -0500

I'm with you guys; I miss the beauty of darkroom photography. I
could carry on at length, especially about digital image
manipulation, but we have to move on (and I'm having a busy Friday
afternoon).

The *short* answer to your immediate question is, using Photoshop, go to
Image - Adjustments - Levels (the first thing in the picklist for a
reason). See histogram. Adjust the white triangle and the black triangle
toward the edges of the histogram, then move the gray triangle in the
middle to where you like the results, usually more toward the peak. If you
still want more contrast *after* this, *then* use Contrast and Brightness.

This is the best thing you can immediately do, plus it's just about all
you can do according to the MSA guidelines on the ethics of digital
imaging, which I can't find on the new, redesigned MSA site.

Aloha,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

==============================Original Headers==============================
5, 23 -- From tina-at-pbrc.hawaii.edu Fri May 14 18:34:34 2010
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From resultskatou0819-at-hanshin-fan.com Fri May 14 18:34:51 2010
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I might add a bit more to what Tina suggests. And I also understand and
appreciate the fine points of wet photography. I have a very good 4K CCD
camera on one of our TEMs and I rarely use it. On the other hand I have not
printed in the darkroom for about 5 years and our last Durst enlarger is on
its way to salvage (let me know if you want it). I capture images on film
and then we scan the film at appropriate dpi for it's intended use (usually
600dpi for must images). I find I can capture better grayscale images on
film faster than if I use the digital camera and and it much easier to
manipulate the contrast gamma, and brightness when scanning to end up with a
very reasonable digital image. However, I also can easily stigmat and focus
on the viewing screen and that is also almost a lost art.

The 15min it takes to develop 30-40 negatives is a minor inconvenience for
the larger and higher quality viewing area. Yes, we then have to put in time
scanning but the final outcome is worth the little extra time.

I suggest you use Photoshop and use "Layer } } } adjustment layer } } } levels"
rather than "image } } } adjustment } } } levels". Using the new adjustment layer
lets you modify the original image and then save the image. This does not
alter the original scanned image but rather saves the changes in a new
layer. Then when you next open the image you can easily readjust, again
without changing the original image. When you finally get the image you
want for a specific purpose you can just flatten the image and do a "save
as" to save the modified image. You still have the original unmodified
image to archive.

Printing is another problem. As we all know, different monitors display
images with slightly different contrast/brightness. Best is to play around
until you get one image correct for your printer and then use that as a
reference when trying to print other images. None will look quite like a
darkroom print but then very few prints are used these days. Most people
just want to study digital images on their computer screen. We almost never
print images anymore. Our users just download the digital images to their
memory stick and away they go....

However, bottom line is that us old timers are fighting a loosing battle.
Digital is here to stay and it does work well for diagnostic images and
other uses that do not require that little extra that you get with film.
Doing tomography without a digital camera is not impossible but almost!
Students want fast and digital gives it to them. Unfortunately they do not
understand what is missing so are happily oblivious to the limitations of
digital imaging.

Debby


---
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy




On 5/14/10 7:36 PM, "Tina Carvalho" {tina-at-pbrc.hawaii.edu} wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} I'm with you guys; I miss the beauty of darkroom photography. I
} could carry on at length, especially about digital image
} manipulation, but we have to move on (and I'm having a busy Friday
} afternoon).
}
} The *short* answer to your immediate question is, using Photoshop, go to
} Image - Adjustments - Levels (the first thing in the picklist for a
} reason). See histogram. Adjust the white triangle and the black triangle
} toward the edges of the histogram, then move the gray triangle in the
} middle to where you like the results, usually more toward the peak. If you
} still want more contrast *after* this, *then* use Contrast and Brightness.
}
} This is the best thing you can immediately do, plus it's just about all
} you can do according to the MSA guidelines on the ethics of digital
} imaging, which I can't find on the new, redesigned MSA site.
}
} Aloha,
} Tina
}
} ****************************************************************************
} * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
} * Biological Electron Microscope Facility * (808) 956-6251 *
} * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
} ****************************************************************************
}
} ==============================Original Headers==============================
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==============================Original Headers==============================
14, 29 -- From dsherman-at-purdue.edu Fri May 14 21:35:08 2010
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From kotka.agency-at-johnnurminen.com Fri May 14 22:39:53 2010
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Greetings all,
The Microscopy and Microanalysis meeting in Portland, OR is a little
more than 2 months away*..

To all students (or those of you who might have students) attending the
meeting, please consider the student bursary program offered by MSA.
The purpose of these bursaries is to encourage students to attend the
annual MSA/MAS Microscopy and Microanalysis meeting, where they can meet
and interact with the established microscopy community while defraying
some meeting costs.

The students work for 20 hours (or up to 40 hours) during the meeting
and pre-meeting events and are paid $10 an hour. The jobs involve such
things as providing support in the different symposia (helping with
audio-visual needs, maintaining an attendance count, and helping
speakers set up for their presentation), staffing the MSA Megabooth or
volunteer office, monitoring use of the Internet Café, and helping with
vendor tutorials and poster set-up.

Once the task list has been finalized, each bursary will be contacted
and allowed to choose the times and activities they would like to work.
Many times they end up *working* sessions they would attend anyway.
There is an added bonus of a $10 cash meal allotment for each morning
and/or afternoon sessions worked.

If anyone would like to participate in the bursary program, please
check the *I wish to apply for a student bursary* box in section 2
of the registration form. Bursary space is limited, so sign-up early.
Applicants for the bursaries must be members of MSA or MAS, and enrolled
as students at a recognized educational institution.

For those *non-students* we could always use volunteers to help
with the above mentioned meeting activities as well. Although not paid
on an hourly basis as the student bursaries, volunteers do receive some
compensation along with the same cash allotment for meals. Plus they
also have the opportunity to interact more with the microscopy community
as they assist with meeting tasks.

If anyone has any questions about the bursary/volunteer program, or
would like to participate, please contact:

Amanda Lawrence
Electron Microscope Center
Mississippi State University
662-325-3019
alawrence-at-entomology.msstate.edu



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10, 20 -- From ALawrence-at-entomology.msstate.edu Sun May 16 06:19:20 2010
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From kristen-at-lcstaffing.com Sun May 16 07:20:45 2010
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Hi all!
 
I worked during my thesis exclusively with films but afterward I worked only with digital cameras, so I am just sitting between 2 chairs!
Simply said I do not regret the films.
 
Electron microscopists are highly specialized/trained scientists and their knowledge and know-how are not easily transmitted and the electron microscopists are proud of it.
In addition to be good in this field, one has to be perfectionist because everything has to be perfect to give something meaningful. Just following protocols is not enough; one has to understand each step in order to adapt it to various specimens.
 
That said, films are not for beginners for various reasons, the most obvious being that one cannot verify the quality of the picture until the final development. So you must be good and have enough experience to choose the right field of view, the right magnification and adjust accurately. I think that this is already a good reason why electron microscopists tend to like films: this is THEIR talent!
Another reason reflected by the message of Robert is that films offer more “wow†effect, they look nicer. But this is related to art, not to science!
 
Fact is: we use our microscopes to extract information, which means contrast in images. And cameras are simply good enough at it. It doesn’t really matter if the contrast is better with films than with cameras, as long as the information is there does it?
As for the field of view, you may need a larger camera but if you don’t have one, there is an alternative: decrease the magnification! You may lose resolution, but you rarely look at details when you take a large field of view. If needed you can simply take other pictures at higher mag. Cameras have the big advantage of being able to give an immediate idea of the result and to collect fast as many images as you like.
One advantage I also appreciate is that my 5 years-collection of images does not weight 50 kg and do not take one entire shelf.
Debby also correctly discussed the need to print on paper (vs. watching on a monitor). Think about how many pictures you take for each you publish!
And we don't even discuss about the ease to make simple statistics like particle counting (and mean diameter determination), which can be done in 30 min with a digital camera even automated.
 and Best regards,

Stephane





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From repetitionsft2-at-rdzeng.com Mon May 17 06:36:47 2010
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On 14/05/2010 18:11, "paul_hazelton-at-umanitoba.ca"
{paul_hazelton-at-umanitoba.ca} wrote:

} There is going to be a need to upgrade the computer and printer hardware
} and software. And I really want to do this right the first time. Which
} leads to the questions
}
} * What programs are people using that provide for better micrograph
} processing,

I manage a couple of TEMs used for brain tissue imaging, one has a Gatan
US1000 (2k bottom mount) and one has a Gatan 1k side-mount camera. I have
trained users in contrast adjustment so that they get OK images for analysis
and quantification, usually using ImageJ software, that can open DM3 files
(including magnification, and keeping the white and black points set in DM).
I usually process images after they have been chosen for publication.

I use Photoshop, I am used to using it, and it allows a lot of freedom. I
just refer to the field of view information (usually taken from ImageJ) when
making scale bars. The key is to understand the bit depth of the camera,
the fact that the image you are seeing is a processed form, and does not
have all the recorded grey values. The lowest black value and the highest
white value are chosen by Digital Micrograph, I think by ignoring a certain
percentage of the image that it thinks are outliers (extreme white pixels
and extreme black pixels). You can set this percentage in one of the menus.
You can also adjust the white and black point (just for the current image)
in a menu, so you can recover parts of the image that were 'lost' (that
don't appear on the histogram in DM). Ask your Gatan contact about this, I
think it is one of the most important this to know about DM.

When you export from DM, you can get a 16-bit file that is exactly what was
digitized (this will appear as grey in Photoshop, you may not even see any
image at all), or you can use the displayed image as the basis of an 8-bit
export- you don't want this version if you want to do any alteration to
contrast. ImageJ can also export as a 16-bit file.

When required, I use non-linear contrast adjustments. I always state that
they have been used, and I apply every process to all parts of the image
(something many were not doing with wet printing). In fact, even the
in-build Digital Micrograph Gamma adjustment is non-linear, it is just a lot
slower for me to get the right image tones using it. I use the curves
adustment in Photoshop almost exclusively.

} * What printers are good. Frankly, the best we have here is not much
} improved over my very old HP 1200 toner printer in the lab. Again,
} as with processing the images, the best I can get is on my home
} system, where I use an HP6400 multifunction system. And I don¹t
} like those prints.

I used to run an Epson A3 inkjet with quad black cartridges from Lyson (you
get ink in different shades of grey to match the reflectance of the coloured
inks, and when the printer uses its built-in colour blending to make greys,
you never get a colour shift, and even pale shades do not have dithering.
You have to use a matched paper, else you get rapid fading and discolouring.

I gave up with it, as it clogged when not in use (like all inkjets in my
experience). So now I don't make nice prints very often, and if I do, I use
an online photo printing service that produces a real photographic print,
and they profile their printer so I know what the image will look like. The
ultimate way of printing is to send a digital file to Ilford, who use a
modern light-scanning digital printer to print onto their black and white
paper. In the UK they have an online photo printing service.

We use a standard HP office laserprinter (4250 I think) in each EM room for
everyday record prints.

Hope that is of some use,


Ben

--
Imaging Technician
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271866
{http://mrcanu.pharm.ox.ac.uk/}



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From scott.nickell-at-sumcousa.com Mon May 17 08:50:05 2010
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Paul;
You did not state what sort of "used Gatan system" you have, and
that may be your fundamental problem. If you have one of the older 1KX1K
cameras with 14 bit depth, the images are going to look disappointing no
matter what software, computer, or printer you use.

John Mardinly
Western Digital

-----Original Message-----
X-from: paul_hazelton-at-umanitoba.ca [mailto:paul_hazelton-at-umanitoba.ca]
Sent: Friday, May 14, 2010 10:12 AM
To: John Mardinly

25 years ago the idea of digital image capture was totally enamouring.
The ability to get rapid prints of clinical and diagnostic material was
fantastic. Even if the prints from the original fibre optic systems were
not to the same resolution, the improvement in Time to Report from
almost a week to less than a day was fantastic. And there was always wet
chemistry for publication prints or if we really needed resolution.

25 years later, as we finally get digital, I find the reality somewhat
less than captivating . But then, I have 40 years of wet chemistry
experience, and at the risk of sounding conceited, I am fairly good at
it. Of course, it could be argued that if you spend 40 years on it, you
should be able to teach a chimpanzee to make a relatively acceptable
print with wet chemistry. So hopefully this is not some closet Luddite
within who is battling his way out.


The problem is - we have had a used Gatan system installed, and are
capturing the images using Gatan DigitalMicrograph software. On the
monitor the images look alright, but just that - alright. Frankly, the
resolution is less than on the focusing screen, but for diagnostic, they
are alright. Hopefully I will be able to figure out how to actually
adjust the different brightness/contrast settings so that we can get
away from averaged optimized data capture, which should improve the
original data.

The system falls apart when it is time to take the micrographs away to
process and print them. I have been using PhotoShop CS4 Enhanced, with
64 bit processing on my computer at home, and am not all that impressed.
Neither does Illustrator CS4 excite me. Ignoring the fine detail
resolution - there is no such thing as a fibre, forget it - the
immediate technical problem is getting acceptable prints. The adjustment
of contrast and brightness seems to be highly limited before bloom
effect takes over (Gee, using British/Canadian idiom, may I call it the
Blooming Effect!!!). There simply is not sufficient gray scale variance
to get a good micrograph. The contrast is too extreme, the background is
whited out due to saturation, etc.


There is going to be a need to upgrade the computer and printer hardware
and software. And I really want to do this right the first time. Which
leads to the questions

* What programs are people using that provide for better micrograph
processing,
* What printers are good. Frankly, the best we have here is not much
improved over my very old HP 1200 toner printer in the lab. Again,
as with processing the images, the best I can get is on my home
system, where I use an HP6400 multifunction system. And I dont
like those prints.



So, thanks in advance. And for my fellow closet Luddites, dont remind
me that it is, after all, a digital world, and the digital world is
[please insert your appropriate scatiological terminology representing
my favourite viral specimen source (fecal)]. We don't have to simply
accept the issues of pixel size, etc. We can do better.


Paul
--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 William Avenue
Winnipeg, Manitoba, Canada, R3E 0J9
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926


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From donarta-at-hakiki.com Mon May 17 11:50:53 2010
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Hi folks.

I recall an interesting discussion recently with a curator of photography at a well known museum
here. She pointed out that the physical nature of a photograph is quite different from a print. In a
print, the ink is deposited in a single layer on the surface of the paper. In photography, both in the
film and the paper, the silver is embedded in a deeper matrix in the paper. The end result may
very well be that "breath-catching" moment when looking at a photographic micrograph, since the
image is formed by a combination of simple reflection and interference within the matrix.

So, beauty is a little more than skin deep.

Joel





On 14 May 2010 at 12:11, paul_hazelton-at-umanitoba.ca wrote:

}
}
}
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} 25 years ago the idea of digital image capture was totally enamouring.
} The ability to get rapid prints of clinical and diagnostic material was
} fantastic. Even if the prints from the original fibre optic systems were
} not to the same resolution, the improvement in Time to Report from
} almost a week to less than a day was fantastic. And there was always wet
} chemistry for publication prints or if we really needed resolution.
}
} 25 years later, as we finally get digital, I find the reality somewhat
} less than captivating . But then, I have 40 years of wet chemistry
} experience, and at the risk of sounding conceited, I am fairly good at
} it. Of course, it could be argued that if you spend 40 years on it, you
} should be able to teach a chimpanzee to make a relatively acceptable
} print with wet chemistry. So hopefully this is not some closet Luddite
} within who is battling his way out.
}
}
} The problem is - we have had a used Gatan system installed, and are
} capturing the images using Gatan DigitalMicrograph software. On the
} monitor the images look alright, but just that - alright. Frankly, the
} resolution is less than on the focusing screen, but for diagnostic, they
} are alright. Hopefully I will be able to figure out how to actually
} adjust the different brightness/contrast settings so that we can get
} away from `averaged´ optimized data capture, which should improve the
} original data.
}
} The system falls apart when it is time to take the micrographs away to
} process and print them. I have been using PhotoShop CS4 Enhanced, with
} 64 bit processing on my computer at home, and am not all that impressed.
} Neither does Illustrator CS4 excite me. Ignoring the fine detail
} resolution - there is no such thing as a fibre, forget it - the
} immediate technical problem is getting acceptable prints. The adjustment
} of contrast and brightness seems to be highly limited before bloom
} effect takes over (Gee, using British/Canadian idiom, may I call it the
} Blooming Effect!!!). There simply is not sufficient gray scale variance
} to get a good micrograph. The contrast is too extreme, the background is
} whited out due to saturation, etc.
}
}
} There is going to be a need to upgrade the computer and printer hardware
} and software. And I really want to do this right the first time. Which
} leads to the questions
}
} * What programs are people using that provide for better micrograph
} processing,
} * What printers are good. Frankly, the best we have here is not much
} improved over my very old HP 1200 toner printer in the lab. Again,
} as with processing the images, the best I can get is on my home
} system, where I use an HP6400 multifunction system. And I don´t
} like those prints.
}
}
}
} So, thanks in advance. And for my fellow closet Luddites, don´t remind
} me that it is, after all, a digital world, and the digital world is
} [please insert your appropriate scatiological terminology representing
} my favourite viral specimen source (fecal)]. We don't have to simply
} accept the issues of pixel size, etc. We can do better.
}
}
} Paul
} --
} Paul R. Hazelton, PhD
} Viral Gastroenteritis Study Group
} University of Manitoba
} Department of Medical Microbiology
} 511 Basic Medical Sciences Building
} 745 William Avenue
} Winnipeg, Manitoba, Canada, R3E 0J9
} e-mail: paul_hazelton-at-umanitoba.ca
} paulhazelton-at-mts.net
} Phone: 204-789-3313 (w);
} 204-489-6924 (h)
} Cell: 204-781-6982
} Fax: 204-789-3926
}
}
} ==============================Original Headers==============================
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--
Joel B. Sheffield, Ph.D.
Biology Department, Temple University
1900 North 12th Street
Philadelphia, PA 19122
jbs-at-temple.edu
(215) 204 8839, fax (215) 204 0486
http://astro.temple.edu/~jbs



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From ksd-at-deitzco.com Mon May 17 13:34:57 2010
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{zaluzec-at-microscopy.com} ; Mon, 17 May 2010 19:34:56 +0100

Hello fellow Microscopists,
I don't do a lot of semi-thin sectioning, but a recent request has led me to the following question:
Why would two (different animals) of the same thick (0.25 um) semi-thin sections show different color hues when stained identically?
The stain is Richardson's (1% methylene blue, 1% azure II, 1% borate) stained for 30 sec on a hot plate then water rinsed and sealed with permount. One of the two sample sets shows more purple than blue? Any thoughts will be greatly appreciated.

M Delannoy

==============================Original Headers==============================
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From john.price-at-sandoz.com Mon May 17 14:31:56 2010
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My first comment is that 0.25 um is pretty thin for a semi-thin. I usually use 0.5. Unless you are carefully selecting sections by their interference colors while floating on water, I doubt they are all uniformly 0.25 um and this might alter staining patterns. In addition, the degree of cross-linking of the resin will alter the staining properties so if there are different batches of plastic resin used for embedding or they were heated down in the oven at different shelf levels and the top of the oven was hotter, you may get differences in staining. 30 sec seems short for staining - I might go longer at a lower temperature to get a more even end result but my experience would suggest it is subtle differences in the resin that caused the difference. Differences in fixation duration might also impact staining. Tom


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: delannoy-at-jhmi.edu [mailto:delannoy-at-jhmi.edu]
Sent: Monday, May 17, 2010 1:47 PM
To: Phillips, Thomas E.

Hello fellow Microscopists,
I don't do a lot of semi-thin sectioning, but a recent request has led me to the following question:
Why would two (different animals) of the same thick (0.25 um) semi-thin sections show different color hues when stained identically?
The stain is Richardson's (1% methylene blue, 1% azure II, 1% borate) stained for 30 sec on a hot plate then water rinsed and sealed with permount. One of the two sample sets shows more purple than blue? Any thoughts will be greatly appreciated.

M Delannoy

==============================Original Headers==============================
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From: mcauliff-at-umdnj.edu
Date: Mon, 17 May 2010 15:11:08 -0500
Subject: [Microscopy] Re: digital micrograph imaging software

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

Greetings all:

I am also a old hand at darkroom work, I even mix my own chemicals from
scratch. Nevertheless, I am digital now due to the huge savings in time.
There are no reasons why you cannot get high-quality prints from
digital. You do need a good photo printer with multiple gray inks, Epson
(and others) make such
machines for less than US$1000. Also Photoshop is a fine tool with many
more features than one really needs. I have no knowledge of the Gatan
system you are using but it could
be the source of your troubles. If you cannot find resolution with CS4
it probably is not there to begin with.
I suggest taking several of your images to a professional photo lab and
get their opinion as to resolution. If the resolution is not there you
are wasting your time and money. If it is there a pro
lab with a good printer will find it.

Geoff

paul_hazelton-at-umanitoba.ca wrote:
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
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}
} 25 years ago the idea of digital image capture was totally enamouring.
} The ability to get rapid prints of clinical and diagnostic material was
} fantastic. Even if the prints from the original fibre optic systems were
} not to the same resolution, the improvement in Time to Report from
} almost a week to less than a day was fantastic. And there was always wet
} chemistry for publication prints or if we really needed resolution.
}
} 25 years later, as we finally get digital, I find the reality somewhat
} less than captivating . But then, I have 40 years of wet chemistry
} experience, and at the risk of sounding conceited, I am fairly good at
} it. Of course, it could be argued that if you spend 40 years on it, you
} should be able to teach a chimpanzee to make a relatively acceptable
} print with wet chemistry. So hopefully this is not some closet Luddite
} within who is battling his way out.
}
}
} The problem is - we have had a used Gatan system installed, and are
} capturing the images using Gatan DigitalMicrograph software. On the
} monitor the images look alright, but just that - alright. Frankly, the
} resolution is less than on the focusing screen, but for diagnostic, they
} are alright. Hopefully I will be able to figure out how to actually
} adjust the different brightness/contrast settings so that we can get
} away from ‘averaged’ optimized data capture, which should improve the
} original data.
}
} The system falls apart when it is time to take the micrographs away to
} process and print them. I have been using PhotoShop CS4 Enhanced, with
} 64 bit processing on my computer at home, and am not all that impressed.
} Neither does Illustrator CS4 excite me. Ignoring the fine detail
} resolution - there is no such thing as a fibre, forget it - the
} immediate technical problem is getting acceptable prints. The adjustment
} of contrast and brightness seems to be highly limited before bloom
} effect takes over (Gee, using British/Canadian idiom, may I call it the
} Blooming Effect!!!). There simply is not sufficient gray scale variance
} to get a good micrograph. The contrast is too extreme, the background is
} whited out due to saturation, etc.
}
}
} There is going to be a need to upgrade the computer and printer hardware
} and software. And I really want to do this right the first time. Which
} leads to the questions
}
} * What programs are people using that provide for better micrograph
} processing,
} * What printers are good. Frankly, the best we have here is not much
} improved over my very old HP 1200 toner printer in the lab. Again,
} as with processing the images, the best I can get is on my home
} system, where I use an HP6400 multifunction system. And I don’t
} like those prints.
}
}
}
} So, thanks in advance. And for my fellow closet Luddites, don’t remind
} me that it is, after all, a digital world, and the digital world is
} [please insert your appropriate scatiological terminology representing
} my favourite viral specimen source (fecal)]. We don't have to simply
} accept the issues of pixel size, etc. We can do better.
}
}
} Paul
}


--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff-at-umdnj.edu
**********************************************





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From: nholson-at-ucsd.edu
Date: Mon, 17 May 2010 15:35:49 -0500
Subject: [Microscopy] Re: digital micrograph imaging software

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

I replied to Paul privately but since this thread is continuing I
decided to write. Printing a digital image is world's different than
just sending a MS Word file to the printer. Also, adjusting the
brightness and contrast is not enough. I have been doing fine art
photography and showing my work in galleries and Photoshop is the
program to use regardless if you are doing color or B&W. I don't
print much of my scientific stuff anymore since papers are submitted
electronically but after Friday's discussion I sent one of my DM3s
(converted to 16 bit grayscale tiff) to my home machine where I have
my Epson 7800, a 24 inch carriage archival pigment inkjet. The
print I made from our 2K Gatan camera looked as good, if not better,
than any of my silver gelatin prints of the past.

I purchased my first high-end printer (Epson 2200) about five years
ago. I was disappointed with the results because they weren't as
good as prints from an older printer I had. I then took a course on
digital printing and I took another more extensive course a few years
ago. Photoshop does a good job of color management that a number of
other software programs don't do (PowerPoint for example). My Epson
7800 has eight different inks (3 blacks if I remember right). A new
Canon I saw advertised in one of my photo rags had 12 inks.
Standard CMYK (4 inks) just doesn't do it because the blacks usually
come out with a color cast. To get a good print you need to match
the printer, the inks and the paper. This is done with a color
profile file that is usually available on the paper manufacturer's
web site. You also need to have a color calibrated monitor and work
in a room with subdued light.

One of the newer Epson printers is the Epson 3880 and it runs around
$1500. I keep mentioning Epson because they had the market until a
few years ago and I haven't kept up with the other brands. These are
inkjets and they need to be babied otherwise you will have problems
but they produce fantastic results.

I have a book on printing that I have read cover to cover. It is
Color Confidence: The Digital Photographer's Guide to Color
Management by Tim Grey. Now that says color but the concepts are
similar. I have absolutely no desire to go back into a chemical
printing darkroom. I say good riddance.

Norm Olson

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

--



______________________________________________________________
Norm Olson
Cryoelectron Microscopy Facilities Manager
1510 Bonner Hall
Department of Chemistry & Biochemistry, MC-0378
University of California San Diego
La Jolla, CA 92093-0378
nholson-at-ucsd.edu
http://cryoem.ucsd.edu
Cell: (858)220-2183
(858)822-6718 - Office; (858)534-5846 - Fax
______________________________________________________________

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From: oshel1pe-at-cmich.edu
Date: Mon, 17 May 2010 15:38:38 -0500
Subject: [Microscopy] Re: LM Slide colors

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

One reason is that methylene blue oxidizes in solution to a mixture
of azure A, azure B, and other thiazine dyes. The mixture is
uncontrolled, but this is done deliberately for some polychrome
staining. What are the differences between your samples? Species,
tissue, ... ?
If just different individuals, and all else is the same, I'd look to
section thickness and subtleties of embedding/fixation and the like.

Phil

} Hello fellow Microscopists,
} I don't do a lot of semi-thin sectioning, but a recent request
} has led me to the following question:
} Why would two (different animals) of the same thick (0.25 um)
} semi-thin sections show different color hues when stained
} identically?
} The stain is Richardson's (1% methylene blue, 1% azure II, 1%
} borate) stained for 30 sec on a hot plate then water rinsed and
} sealed with permount. One of the two sample sets shows more purple
} than blue? Any thoughts will be greatly appreciated.
}
} M Delannoy

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: sbras-at-uw.edu
Date: Tue, 18 May 2010 00:23:48 -0500
Subject: [Microscopy] viaWWW: SEM Back-scatter Detector

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

Dear listers,

Perhaps you'll remember my issue with our Tecnai G20 several weeks ago.
I first want to thank all those of you who took time to give me advices.
It took time to solve the issue, mainly because the problem arose any time, sometimes we had to test a solution for 2-3 days.
The issue is now solved and it is a great relief, esp. because this will not cost too much.
I just wanted to share the solution with everybody, so hopefully you'll come to the idea to test this piece if it ever happens to you too.
First of all we needed to bypass the interlocks to make sure that this was not the cause for the microscope to switch off.
But the problem arose even with all interlocks bypassed so they could not be the cause.
Then someone from FEI came to the idea that sometimes a 24V boards fails in their system so we tested the current and we saw that the voltage changed when the TEM switched off. We changed the board and now our TEM works!
I want to aknowledge the professionalism from the FEI personal in Europe, who did their best to provide help and support and to solve the problem while avoiding unnecessary costs.

Regards,

Stephane



----- Original Message ----
X-from: "nizets2-at-yahoo.com" {nizets2-at-yahoo.com}
To: nizets2-at-yahoo.com
Sent: Wed, March 24, 2010 10:19:15 AM

Dear all,

I am having trouble with our Tecnai G20/SF6. I start the microscope and all is fine, it is ready for action.
I start the HT and I let stabilize for 1 hour. I control the chillerwater temperature and it is stable (17-18°C).
Really all looks perfectly normal.
Then I leave the microscope to go and eat at noon and when I come back the display is frozen, nothing works and the microscope is OFF (the main lamp is on OFF).
I start the PC and the microscope and again everything is fine. I leave it for 2 hours, until I go home and all looks fine.

I come back the next day in the morning the display on the monitor is frozen and the microscope is OFF!!!
I started it again and all looks fine until now, I am controlling everything regularly but in the meantime I thought I would send a message in the bottle to the list to know what you think about this story.

Now some side notes:

- The emission chamber has been filled with SF6 on monday up to 6 bar. I had the time (fortunately) to condition the HT just before the error happened. Actually I went all up to 220kV then I left the microscope for the noon pause on 220kV and when I came back all was frozen (microscope, vacuum and HT off).

- There was a recent change in the water chiller: the water was always set to 15°C in charge and the temperature limit (backflow) was set on 24°C. 2 weeks ago the machine stopped working. When I investigated, I set the temperature limit to 30°C and it started again, the water temperature reaching 19°C in charge (without changing the temperature setting). I let it go because these values are OK I think.

- On the "frozen display", there is not a single error message, nothing. It is just as it was when I left it.

- I controlled the log file and, although I understand almost nothing from the hundreds of messages, it seems that no error occured. There are just info messages, no error messages.

- One strange thing: when the vacuum in the column is broken, there is a red LED lighting on the goniometer. When the problem happens and the display is frozen and the microscope is OFF, the red LED is not lighting. However when I start the system new all the vacuum are broken. I don't understand how the red LED was not lit while the vacuum was broken.

I have 2 hypotheses: maybe it is the chiller, but I wonder how it can work fine for 3 hours and then suddenly fail, then work again and so on...
Or it has to do with the PC, which controlls everything. Something in the PC is unstable and crashes and this crashes the whole system. Unfortunately I know too few in computers to investigate this issue myself.

I would be glad if you'd care to share your thoughts about this issue with me.

regards,

Stephane






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From kovnicek-at-spokaneindustries.com Mon May 17 16:13:16 2010
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Well folks

I tried to sit this one out because this might end up being very long
and I just don't type that well: but
First: Digital Micrograph is a great program but not for printing. For
printing you want Photoshop. But you can use almost any version since
Photoshop 5. The trick is simply to understand how to print and what to
print on. Best B?W will be Epson for so many reasons and three major
patents that it isn't worth talking about. All pro photographers use
Epson printers We have tested printers for years now and the Epsons are
clearly superior. The best current model is the Epson 2880. It will
print up to 13"x17" which covers everything except posters (We use an
Epson 9600) The K3 (3 blacks are important because you can print at
higher densities than 300 ppi. This is because the dot pattern can be
denser if you have gray inks. Gray inks are very important for the best
but you can do very well with a C88 plus on good paper. The C88 plus
printer costs $85. The 2880 is about $800. The C88 plus prints the best
images on plain paper.
We have always compared our prints (and still do ) to the same negative
printed on Agfa Brovira paper. If you have enough pixels then the prints
will be very close. The photograph will be sharper but only to a trained
observer. Now I have heard all the arguments about resolution and pixel
size but for a biologist there is an important and rather simple
calculation that I think defines the problem. The old photo prints were
8" by 10 ". A good digital print is about 600 ppi. The simple math
says 4800 x 6000 pixels. That is going to take a rather large montage to
equal with a smaller digital camera. So you either need a larger format
camera, a large number of montage images or record on film and scan.
4k cameras are out of my laboratory's budget but a $700 scanner can do
2000 ppi (we have calibrated it) Epson wouldn't you know. ( D750-M pro
or V700)(also tested 4880 and 4990) It is quite easy to get enough
pixels by scanning at 1200 ppi and then we are totally digital. When you
want the best you can go up to 6000 x 8000 pixels.
in one pass!
Now I don't have alot of experience with conversion of images from
Digital Micrograph but you really never want software to decide on
limits for you. This is especially true if it is trying to get a larger
range by binning values in the white and the black. This will produce
awful prints. You want to follow what Tina outlined and use levels in
Photoshop. ImageJ is a great image processing program but doesn't print
very well at all.
Levels is important. Photoshop uses the center control of levels to
adjust the gamma. The gamma function is a curve that is defined. If you
use curves you are drawing a function freehand. Photoshop has no way
that I know of of reading out that curve. I don't think a lot of
journals would allow you to state that you drew a graph freehand. The
gamma function is a log function. Guess what else responds by a log
function. Human vision for one and display monitors for two. So you want
to use gamma for prints so they look right also. That is also why most
find it easy to adjust images on screen and not for printer. Without
adjusting gamma it really won't work very well. You should also use
gamma adjustment for adjusting images for display.
Now here is an important point. You can never know what another person's
monitor will be like so there is really no standard for adjustment. Just
try it on a couple of "average" computers and that's the best you can
do. The gamma for every display is different. That's why projectors make
everything look so bad. We now bring our own projector if it is critical.
Now it was suggested that color correction is the way to travel with
correction and I could not disagree more. I would refer you to Bruce
Fraser's book on Color correction. While he was alive he was the guru
that Adobe listened to. He points out the severe limitations of color
correction and I have found that for scientific printing it does not
help. This is especially true for black and white (which has no color ro
correct). The diimmed light environment needed can be difficult to
achieve. (The dimmable fluorescent lights in my office cost $2,800 so I
know from experience. Not to mention all the wise cracks I get about
working in a darkened cave) The worst part is that to calibrate a
monitor (CRT or LCD) you must turn the intensity to a linear range.
That means you must reduce the brightness to 50% (For reference most
monitors run at 95% or higher). Even in a dim room, you can have trouble
working.
So the procedure we follow is as Tina said

Levels
Adjust histogram till black arrow and white arrow just touch. Then print.
adjust gamma by at least 1.2 print again
repeat until it is clear you have too much gamma
Why? cause gamma is like focus you don't know that you have too much
until you go past it.
After you have chosen the best gamma value you do a final histogram
stretch to reclaim the contrast. We do this selectively t in the dark
region of the histogram where there is little data that will be masked
out (lost). We can reclaim a huge amount of contrast while binning less
than a couple of percent of the data

The best histogram to start with is one that has no values in the dark
or white regions (about 15-20 values) this insures the image is neither
over or under saturated.
When scanning, we scan in 16 bit mode then do histogram stretch in 16
bit mode in photoshop before converting to 8 bit mode for gamma
adjustment and printing. (Note no print driver known can handle native
16 bit images) Then proceed with Levels adjustment.as outlined

With a good image, we can get publishable quality prints on any Epson on
any paper.
Secondly, you are not doing anything that is not scientifically correct.
(Unless you publish in the one Journal that does not understand gamma
and thinks it is wrong)
I hope I have included enough for this to make sense. There are really
a lot of topics covered and felt I should throw in my two cents.
One of the problems going forward is that scientific digital imaging is
far more restrictive than digital imaging in general and new students
are going to have to unlearn what they think they can do.
As a final note to those who miss photography and photographic resolution.
When the quality got good enough I was only too glad to switch. 3-5
hours in the darkroom was fun when I was a graduate student and was
tedious by the time I was a postdoc. It only became tolerable again when
I had a technician to do it for me. Now we get results in minutes.
If you are not satisfied with the quality then just wait. I've been
using computers since 1965
When I started doing digital imaging in 1982 on an s100 bus computer
running CPM, I was creating these huge 262k files (512 x 512 pixels) My
cheap Sony one shot now has a 12 mega pixel sensor and produces images
that are 36 megapixel. If you look at the dot pattern of the Epsons,
they mimic the photograph emulsion but the dots are, of course, much
bigger than grains. If the digital imaging companies can make the images
clearer and since the storage is free, why wouldn't they? So just wait
soon the cameras on TEMs will be dwarfing the puny 16k by 16k of film.

Just wait it really didn't take us very long to get to here.


John

John M. Mackenzie, Jr. PhD
Coordinator, Center for Electron Microscopy
Professor, Department of Microbology
North Carolina State University
Voice (919) 515-2664 Fax (919) 515-8293

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From toby.rouse-at-lovells.com Mon May 17 23:41:43 2010
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Email: sbras-at-uw.edu
Name: Scott

Organization: University of Washington

Title-Subject: [Filtered] SEM Back-scatter Detector

Message: We are in the market for a back-scatter detector to upgrade
our FEI Sirion SEM. The FEI Centaurus detector is more expensive
than others available, but I wonder if it has some significant
advantages. Does anyone have experience, positive or negative, using
the Centaurus with back-scatter tip? Or do you have any
recommendation for another type or model of back-scatter detector for
geology?

Thanks

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From: underwoo-at-u.washington.edu
Date: Tue, 18 May 2010 12:36:03 -0500
Subject: [Microscopy] Re: Cryo-cutting polymers

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Email: dhorne-at-interchange.ubc.ca
Name: Derrick Horne

Organization: UBC BioImaging Facility

Title-Subject: [Filtered] quantitative EBIC

Message: Not working in a materials lab always raises some
interesting questions when our non-biology counterparts show up. I
have recently had a request for EBIC, actually quantitative EBIC, on
cross sections of catalyst-infused membranes, using our S4700 FESEM.

Having looked into it a little we have determined the cost of an OEM
unit to do this is prohibitively expensive. Does anyone have
experience building a unit to perform the task?

Failing that, would anyone have any insight on where our client could
take or send samples to perform this kind of work? I know he wouldn't
mind going to Europe, but I think the West Coast of North America
would be a better target.

Thanks in advance.

Derrick

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From joerice-at-t1experts.com Tue May 18 03:46:40 2010
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Hi

I am afraid in this world you "get what you pay for" and that generally
means the more they cost the better they are! I am sure others will point
out that BSE detectors are general purpose, but the better their signal
level the easier it will be for the geologist to discriminate the average
density of materials in their specimen.

You should be aware that the top of the range instruments from the major
manufacturers have very advanced backscattered electron detection systems
using the "standard" detectors. This type of detection is pretty
spectacular offering 100,000X plus down to 1kV!

Good luck with your purchase.

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



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Organization: University of Washington

Title-Subject: [Filtered] SEM Back-scatter Detector

Message: We are in the market for a back-scatter detector to upgrade
our FEI Sirion SEM. The FEI Centaurus detector is more expensive
than others available, but I wonder if it has some significant
advantages. Does anyone have experience, positive or negative, using
the Centaurus with back-scatter tip? Or do you have any
recommendation for another type or model of back-scatter detector for
geology?

Thanks

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Dear All,
I just had my first trial run to dry-cut polymers (ABS) at temperatures of -40 deg C for the knife and -25 degress for the
specimen and in a second run with -75 / -40 degrees on glass-knifes.
Finally I got some usable cuts which had been electron transparent :-)

But: The problem I still have, even with the cutting area formed like a two triangles standing on base to each other each cutting
stroke will produce a rolled up cut, like a tube...
Is there a chance to spread the cuts in water or a special solution after picking up?
Do you have any links to share for a beginner in cryo-cutting polymers?


Thanks,
Stefan


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Hi All,

I have cryo thin sectioned polyHEMA and silicone and found that once I get below the glass transition temperature for each (-90 and -110C respectively) the sections start to behave better. But at this point it is the cutting speed that will control some of the curl. It seems that faster is sometimes better } 1mm/sec. You must have a anti-static device as well.

Robert Underwood
University of Washington
Dermatology






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From: bob.price-at-uscmed.sc.edu
Date: Mon, 24 May 2010 14:59:34 -0500
Subject: [Microscopy] viaWWW: Basic Confocal Microscopy Workshop

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

This thread has been interesting, and I would like to add to comments made
by John.

When moving the white and black sliders in the Levels box within Photoshop,
it is sometimes difficult to see where the histogram ends because so few
pixels make up the darkest and brightest parts of the image (on the other
hand, the Photoshop-mimicking free program, GIMP, provides a log scale
histogram so that every pixel tone can be seen).

Here's a tip: if you want to see the parts of the image with the darkest
and brightest pixels, hold down the alt key (PC) or the option key (Mac)
while moving the sliders. For the black slider, the image will turn white
the moment you move the slider; for the white slider, the image will turn
black. Move the slider until significant parts of the image appear, then
back off until these disappear. Note the word "significant:" some parts of
the image may be detritus or artifact. When these parts of the image
appear, it tells you that a move of the slider any closer to the center of
the histogram will result in setting the darkest and whitest parts of the
image outside the dynamic range of the image (clipped or saturated values),
which is okay for known, non-significant parts of the image, but not okay
for significant parts as it will block detail.

No one has addressed the issue of sharpening. I've noticed that some
journals discourage sharpening, but since this thread speaks to printing to
hard copy, sharpening may be an option for bringing out local contrast to
better reveal details. Some call this "enhancement" or "doctoring" while
others call it "resolution." My thought is that any improvement in
revealing detail is something that should be pursued: over-sharpening, or
sharpening to the point at which artifact is confused with
biology/materials must be avoided.
A way to provide local contrast consists in duplicating the background
layer in Photoshop to make a layer above the first layer (Layer} Duplicate
Layer). Then use the High Pass filter (Filter} High Pass) and set the level
between 1 and 4 to the point at which edges are visible but not so much the
features. Set the Layer mode (in the Layers palette) to Hard Light and the
image is sharpened. By clicking on the eye icon (in the Layers palette) to
turn off and on the layer, one can see the subtle effect of sharpening and
consequent improvement in local contrast. This effect works much like
selenium toning on photographs to improve acutance.

Sharpening should be done before setting tonal levels, and to a duplicated
image, not the original or raw image.

Finally, in my own experience, the blackest tones on photographic paper
exceeds that of inkjets, and the darkness of the black tones has a marked
effect on the perception of contrast. Thus the necessity for more than one
shade of black ink in an inkjet printer, as John had mentioned, and the
necessity for sending out to an agency as mentioned in an earlier post. At
one time the U of Minnesota core facility owned a Fuji Pictrography 3000,
which is a dye-sublimation/photographic processing device. This could
achieve nearly the black tones provided on Kodak RC paper, and side-by-side
comparisons with photographic paper could, in some instances, be perceived
as "better" than the wet-processed papers, depending on the sample.

A means for providing a greater perception of contrast can also be done by
blue-shifting the image. As microscopists, we develop a "color memory."
Even though photographic paper is "black and white," the Agfa papers tended
to be bluer than the Kodak papers (which looked more brown under artificial
light), and we became not only accustomed to seeing that color shift, but
became judges of quality based upon the color. Try it sometime: make the
image into RGB Color, and then use the gamma slider in Levels to adjust the
blue channel, and then print.

An anecdote to end this missive: Once I was attempting to convince a
Radiologist to photograph his x-ray films back in the "early" days of
digital imaging. I photographed a long film with a digital camera and
emailed the image. He wrote back to say the image was awful and low in
resolution. I knew that it wasn't a "resolution" problem, that it had to be
something else. I noticed that the x-ray films were blue in color, and so I
took the same image and shifted its color to match that of the xray film
and emailed it to him again. He wrote back and said, "I don't know what you
did, but the resolution is now excellent." It was purely his color memory.

Jerry Sedgewick









john_mackenzie-at-ncsu.edu wrote:
}
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} Well folks
}
} I tried to sit this one out because this might end up being very long
} and I just don't type that well: but
} First: Digital Micrograph is a great program but not for printing. For
} printing you want Photoshop. But you can use almost any version since
} Photoshop 5. The trick is simply to understand how to print and what to
} print on. Best B?W will be Epson for so many reasons and three major
} patents that it isn't worth talking about. All pro photographers use
} Epson printers We have tested printers for years now and the Epsons are
} clearly superior. The best current model is the Epson 2880. It will print
} up to 13"x17" which covers everything except posters (We use an Epson
} 9600) The K3 (3 blacks are important because you can print at higher
} densities than 300 ppi. This is because the dot pattern can be denser if
} you have gray inks. Gray inks are very important for the best but you can
} do very well with a C88 plus on good paper. The C88 plus printer costs
} $85. The 2880 is about $800. The C88 plus prints the best images on plain
} paper.
} We have always compared our prints (and still do ) to the same negative
} printed on Agfa Brovira paper. If you have enough pixels then the prints
} will be very close. The photograph will be sharper but only to a trained
} observer. Now I have heard all the arguments about resolution and pixel
} size but for a biologist there is an important and rather simple
} calculation that I think defines the problem. The old photo prints were
} 8" by 10 ". A good digital print is about 600 ppi. The simple math says
} 4800 x 6000 pixels. That is going to take a rather large montage to equal
} with a smaller digital camera. So you either need a larger format camera,
} a large number of montage images or record on film and scan.
} 4k cameras are out of my laboratory's budget but a $700 scanner can do
} 2000 ppi (we have calibrated it) Epson wouldn't you know. ( D750-M pro or
} V700)(also tested 4880 and 4990) It is quite easy to get enough pixels by
} scanning at 1200 ppi and then we are totally digital. When you want the
} best you can go up to 6000 x 8000 pixels.
} in one pass!
} Now I don't have alot of experience with conversion of images from
} Digital Micrograph but you really never want software to decide on limits
} for you. This is especially true if it is trying to get a larger range by
} binning values in the white and the black. This will produce awful
} prints. You want to follow what Tina outlined and use levels in
} Photoshop. ImageJ is a great image processing program but doesn't print
} very well at all.
} Levels is important. Photoshop uses the center control of levels to
} adjust the gamma. The gamma function is a curve that is defined. If you
} use curves you are drawing a function freehand. Photoshop has no way that
} I know of of reading out that curve. I don't think a lot of journals
} would allow you to state that you drew a graph freehand. The gamma
} function is a log function. Guess what else responds by a log function.
} Human vision for one and display monitors for two. So you want to use
} gamma for prints so they look right also. That is also why most find it
} easy to adjust images on screen and not for printer. Without adjusting
} gamma it really won't work very well. You should also use gamma
} adjustment for adjusting images for display.
} Now here is an important point. You can never know what another person's
} monitor will be like so there is really no standard for adjustment. Just
} try it on a couple of "average" computers and that's the best you can do.
} The gamma for every display is different. That's why projectors make
} everything look so bad. We now bring our own projector if it is critical.
} Now it was suggested that color correction is the way to travel with
} correction and I could not disagree more. I would refer you to Bruce
} Fraser's book on Color correction. While he was alive he was the guru
} that Adobe listened to. He points out the severe limitations of color
} correction and I have found that for scientific printing it does not
} help. This is especially true for black and white (which has no color ro
} correct). The diimmed light environment needed can be difficult to
} achieve. (The dimmable fluorescent lights in my office cost $2,800 so I
} know from experience. Not to mention all the wise cracks I get about
} working in a darkened cave) The worst part is that to calibrate a monitor
} (CRT or LCD) you must turn the intensity to a linear range. That means
} you must reduce the brightness to 50% (For reference most monitors run at
} 95% or higher). Even in a dim room, you can have trouble working.
} So the procedure we follow is as Tina said
}
} Levels
} Adjust histogram till black arrow and white arrow just touch. Then print.
} adjust gamma by at least 1.2 print again
} repeat until it is clear you have too much gamma
} Why? cause gamma is like focus you don't know that you have too much
} until you go past it.
} After you have chosen the best gamma value you do a final histogram
} stretch to reclaim the contrast. We do this selectively t in the dark
} region of the histogram where there is little data that will be masked
} out (lost). We can reclaim a huge amount of contrast while binning less
} than a couple of percent of the data
}
} The best histogram to start with is one that has no values in the dark
} or white regions (about 15-20 values) this insures the image is neither
} over or under saturated.
} When scanning, we scan in 16 bit mode then do histogram stretch in 16
} bit mode in photoshop before converting to 8 bit mode for gamma
} adjustment and printing. (Note no print driver known can handle native 16
} bit images) Then proceed with Levels adjustment.as outlined
}
} With a good image, we can get publishable quality prints on any Epson on
} any paper.
} Secondly, you are not doing anything that is not scientifically correct.
} (Unless you publish in the one Journal that does not understand gamma and
} thinks it is wrong)
} I hope I have included enough for this to make sense. There are really a
} lot of topics covered and felt I should throw in my two cents.
} One of the problems going forward is that scientific digital imaging is
} far more restrictive than digital imaging in general and new students are
} going to have to unlearn what they think they can do.
} As a final note to those who miss photography and photographic resolution.
} When the quality got good enough I was only too glad to switch. 3-5
} hours in the darkroom was fun when I was a graduate student and was
} tedious by the time I was a postdoc. It only became tolerable again when
} I had a technician to do it for me. Now we get results in minutes.
} If you are not satisfied with the quality then just wait. I've been
} using computers since 1965
} When I started doing digital imaging in 1982 on an s100 bus computer
} running CPM, I was creating these huge 262k files (512 x 512 pixels) My
} cheap Sony one shot now has a 12 mega pixel sensor and produces images
} that are 36 megapixel. If you look at the dot pattern of the Epsons, they
} mimic the photograph emulsion but the dots are, of course, much bigger
} than grains. If the digital imaging companies can make the images clearer
} and since the storage is free, why wouldn't they? So just wait soon the
} cameras on TEMs will be dwarfing the puny 16k by 16k of film.
}
} Just wait it really didn't take us very long to get to here.
}
}
} John
}
} John M. Mackenzie, Jr. PhD
} Coordinator, Center for Electron Microscopy
} Professor, Department of Microbology
} North Carolina State University
} Voice (919) 515-2664 Fax (919) 515-8293
}
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--
Jerry (Gerald) Sedgewick
Author: "Scientific Imaging with Photoshop: Methods, Measurement and
Output."
Sedgewick Initiatives
965 Cromwell Avenue
Saint Paul, MN 55114
651-788-2261
jerrysedgewick-at-gmail.com
http://www.quickphotoshop.com
http://www.rawlight.com
http://www.jerrysedgewick.com



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10, 24 -- From zaluzec-at-microscopy.com Wed May 19 00:31:20 2010
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From hmp-at-africa-business.com Wed May 19 00:51:46 2010
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Hello out there,
is there any literature available comparing the P47 and the luminix
scintillators?
What are your thoughts concerning this issue? Can the luminix help
enhance imaging in sem?
Thank you for any comments
Regards
Gerd

--
Gerd Mueller von der Haegen
BIOLAB Umweltanalysen GmbH
Ernst-Boehme-Str. 30
38112 Braunschweig
tel: +49-(0)531-313000
fax: +49-(0)531-313040

gerd-at-biolab.de
www.biolab.de



==============================Original Headers==============================
5, 19 -- From gerd-at-biolab.de Wed May 19 03:45:02 2010
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From koali-at-mail4y.com Wed May 19 04:02:48 2010
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{microscopy-at-microscopy.com} ; Wed, 19 May 2010 13:32:46 +0430

Gerd,
Take a look at http://www.semicro.org/scintillators.aspx
It looks to me like the Luminix is very similar to P47 except for peak
emission and cost. Unless your scintillator is extremely difficult to
change or your PMT is centered at 420nm, I don't see any great advantage.
In fact, all of the single crystal scints need to be reconditioned from time
to time. How that compares to changing a P47 once or twice a year, I don't
know.

Also, some detectors require the scint disk to be a certain thickness to be
held correctly and the 1mm thickness might not work, depending on your
detector.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: gerd-at-biolab.de [mailto:gerd-at-biolab.de]
Sent: Wednesday, May 19, 2010 4:48 AM
To: kenconverse-at-qualityimages.biz

Hello out there,
is there any literature available comparing the P47 and the luminix
scintillators?
What are your thoughts concerning this issue? Can the luminix help
enhance imaging in sem?
Thank you for any comments
Regards
Gerd

--
Gerd Mueller von der Haegen
BIOLAB Umweltanalysen GmbH
Ernst-Boehme-Str. 30
38112 Braunschweig
tel: +49-(0)531-313000
fax: +49-(0)531-313040

gerd-at-biolab.de
www.biolab.de



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From brfyklo-at-spaceshots.com Wed May 19 08:26:15 2010
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Fellow Microscopists,
Thanks to all of you who responded to my question so quickly, I received many helpful insights and suggestions. I would like to summarize for you the responses.

1. Methylene Blue oxidizes in a mix of Azure A and B and other thiazine dyes. It is uncontrolled a my occur over time.
2. pH diferences of the stain, if the sections are of appreciable volumes, pH differences of the two sections may have altered the pH of the stain.
3. Differences in pre and post staining drying times and position on the hot plate.
4. Permount contains organic solvents which elute basic stains from sections.

I overcame everything by matching the interference colors, staining as soon as the sections dried, (30 sec staining), rinse , dry and controlled position on the hot plate. I mounted immediately in permount, THEN RECORDED THE IMAGES IMMEDIATELY. This seemed to work
perfectly. Again thanks to all of you for your immediate responses.

Michael Delannoy


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From doubina-at-funbox3d.com Wed May 19 11:02:21 2010
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Email: Wadowska-at-upei.ca
Name: Dorota Wadowska

Organization: Atlantic Veterinay College at UPEI

Title-Subject: [Filtered] TEM turbo molecular pump

Message: Hello,
Recently we installed a new turbo molecular pump (Edwards STP-301) on
our H7000. I noticed that the speed fluctuates a bit (1%). Is this
something that I should be concerned about or is this acceptable?
TIA
Dorota

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==============================Original Headers==============================
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From managingeditor-at-buzzflash.com Wed May 19 11:49:52 2010
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Dear list,

I know there are free softwares for TEM tomography around. Is there any free 3-D SEM software around too?

Thank you,
Larry

==============================Original Headers==============================
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From donnahulett-at-coldwellbanker.com Wed May 19 13:29:57 2010
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Dear All,

We have the good fortune to run a 300kV FEGTEM with a Mollenstedt-Ducker
biprism that has served us well for quite a few years (about 8). This has
performed well up until recently where we have noticed that the biprism now
starts to charge for even the smallest of beam currents.

Before now, the unbiased biprism would go into the beam and create a nice
dark shadow image (out of focus, of course); now, it creates a thin bright
line indicating some beam overlap caused by some residual voltage. Empty
holograms taken with different spot sizes give different fringe spacings
too (the beam current goes down by a factor of 3-4 between each
measurement):

Spot size Fringe spacing (angstrom)
1 2.44
3 2.55
5 2.67
7 2.75
9 2.82

The effect appears to worsen towards the middle of the biprism, suggesting
that the biprism cannot discharge rapidly enough.

My questions are these. Has anyone had this problem before? If so, what
other tests did you do to confirm that it is the wire itself and not, say,
the biprism voltage supply? Does a biprisms ability to discharge worsen
over time? Is this an oxide layer that thickens as the biprism ages
(despite sitting in high vacuum)?

Thanks.

Jon Barnard


==============================Original Headers==============================
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From kolshire-at-hobbittown.com Fri May 21 04:24:43 2010
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Jon:

8 years? And you're complaining?? ;-)

You need to either re-coat your biprism with something conductive (e.g. sputtered Ir, Au, or the like), or make a new fiber.

e-mail me offline for more details, if you'd like...or maybe we can do a Skype call?

regards,

Larry


Dr. Lawrence F. Allard
Distinguished Research Staff Member
High Temperature Materials Laboratory
Microscopy Group
Materials Science and Technology Division
Oak Ridge National Laboratory
1 Bethel Valley Road
PO Box 2008
Oak Ridge, TN 37831-6064
(note: the last 4 lines are sufficient for mailing or overnight courier service)
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________________________________________
X-from: jsb43-at-cam.ac.uk [jsb43-at-cam.ac.uk]
Sent: Friday, May 21, 2010 4:59 AM
To: Allard Jr, Lawrence Frederick

Dear All,

We have the good fortune to run a 300kV FEGTEM with a Mollenstedt-Ducker
biprism that has served us well for quite a few years (about 8). This has
performed well up until recently where we have noticed that the biprism now
starts to charge for even the smallest of beam currents.

Before now, the unbiased biprism would go into the beam and create a nice
dark shadow image (out of focus, of course); now, it creates a thin bright
line indicating some beam overlap caused by some residual voltage. Empty
holograms taken with different spot sizes give different fringe spacings
too (the beam current goes down by a factor of 3-4 between each
measurement):

Spot size Fringe spacing (angstrom)
1 2.44
3 2.55
5 2.67
7 2.75
9 2.82

The effect appears to worsen towards the middle of the biprism, suggesting
that the biprism cannot discharge rapidly enough.

My questions are these. Has anyone had this problem before? If so, what
other tests did you do to confirm that it is the wire itself and not, say,
the biprism voltage supply? Does a biprisms ability to discharge worsen
over time? Is this an oxide layer that thickens as the biprism ages
(despite sitting in high vacuum)?

Thanks.

Jon Barnard


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From rferguson-at-fsb-law.com Fri May 21 10:32:42 2010
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Dear All,

We have an immediate opening for a permanent staff position as a
scanning electron microscopist in the Electron Probe Instrumentation
Center (EPIC) housed within the NU/ANCE/ Center at Northwestern
University in Evanston, IL. We are seeking someone with a strong
physical science background and some microscopy experience who enjoys
working with people from diverse backgrounds and disciplines.

Please visit: {http://www.nuance.northwestern.edu/jobs.asp} for more
information on this and other open positions in our facility.

Regards,
Ben

--
*Ben Myers*
SEM/FIB Facility Manager
NU/ANCE/ Center

Northwestern University
Mail: 2036 Cook Hall
Office: 1114 Cook Hall
2220 Campus Drive
Evanston, IL 60208-3108

ph: (847) 491-3439
fax: (847) 467-6573

http://www.nuance.northwestern.edu

==============================Original Headers==============================
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From reservations-at-lowellhotel.com Fri May 21 11:17:28 2010
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There has recently been a long thread comparing film with digital
imaging in the electron microscope. Unless I have missed it, this
discussion has missed one of the most important differences between the
two methods of recording images. Film is not a linear medium. In
other words the optical density of film is not proportional to the
exposure. On the other hand, digital recording devices (CCD cameras and
the like) are linear right up to saturation (as far as I am aware).
This means that for any case where the intensity in an image is to be
used for scientific, rather than aesthetic, purposes, digital images are
greatly to be preferred.

A further complication arises in that the use of film in the SEM is not
the same as the use of film in the TEM. In the SEM the film is exposed
to light from a phosphor, and responds in the same way as film used for
conventional light photography. In this case, several photons are
needed to cause a grain to become exposed (developable). In the TEM the
electrons fall directly onto the film. One electron at these energies
is enough to cause a grain to become developable. The response in the
two cases is quite different. In the SEM the response is far from
linear at both low and high exposures, and linear for only a very small
region of moderate exposure. In the TEM by contrast, the exposure is
linear only at low exposures.

Why is it then that so many people find film images more pleasing than
digital images. I can only suppose that there is some form of
correspondence between the non-linearity of the human eye and the
non-linearity of film. So we are left with the challenge of finding a
digital filter that can convert digital images into a form that is more
attractive. The closest I have seen to achieving this are two filters
that do very much the same thing: filters that remove low spatial
frequencies and "unsharp mask" filters. Perhaps someone can devise
something better.

There may be something newer, but the reference I use for understanding
the exposure of film is:
The response of photographic emulsions to electrons,
by R. C. Valentine.
It is in Advances in Optical and Electron Microscopy Volume 1, Edited
by R. Barer and V. E. Cosslett (Academic Press, London 1966) pages 180-203.

--
Alwyn Eades
Lehigh University


==============================Original Headers==============================
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From reneel-at-wesli.com Fri May 21 17:19:27 2010
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Hi,

We would like to invite you to the third Workshop on Electron Crystallography of Membrane Proteins, which will take place August 1-7, 2010, in C-CINA, University Basel, Switzerland.

Aims
The workshop on membrane protein 2D crystallization and electron crystallography is aimed at everybody (PhD students, PostDocs, group leaders, etc.), who is interested in the structure determination of membrane proteins (or other 2D protein crystal arrays) by electron crystallography.

Topics
This workshop is limited to 30 participants, and has two major aims:
- Advancing the technology: To bring together many of the leading groups in electron crystallography to forge innovative collaborations for new technology development.
- Training: To provide a unique forum to train skilled biochemists and electron microscopists (PhD, Post docs and beyond), in membrane protein 2D crystallization, electron crystallographic data collection and data processing.
-- Membrane protein solubilisation and 2D crystallization
-- Sample preparation for electron microscopy
-- Cryo-EM data collection (imaging and electron diffraction)
-- Data processing with the MRC, IPLT and 2dx software packages
-- Data evaluation and model building

Workshop format
The workshop will feature lectures in the morning, practicals in the afternoon, and science talks in the evening. Membrane protein crystallization trials will introduce lipid monolayer crystallization methods, or 2D crystallization with dialysis devices, or with a cyclodextrin robot. We will demonstrate automated 2D crystal screening, and present cryo-EM imaging and electron diffraction on the CM200FEG and Titan Krios. We will lecture image processing using MRC, IPLT and 2DX software systems. Computer practicals will use machines in a local computer room. However, students are also invited to bring their own laptop computer (Linux or OSX), where we can try to install the image processing software packages and run the processing on your own computers during the workshop.

Speakers:
- Anchi Cheng, Scripps, La Jolla (confirmed)
- Andreas Engel, C-CINA, Basel (confirmed)
- Andreas Schenk, Harvard Med. School, Boston (confirmed)
- Ansgar Philippsen, Shaw, NY (invited)
- Bryant Gipson, C-CINA, Basel (confirmed)
- Carsten Sachse, EMBL, Heidelberg (confirmed)
- Catherine Venien-Bryan, Univ. Oxford, UK (confirmed)
- Clemens Schulze-Briese, PSI, Villigen (confirmed)
- Daniel Levy, Inst. Curie, Paris (confirmed)
- Gebhard Schertler, Biology, PSI, Villigen (confirmed)
- Hans Hebert, Karolinska, Stockholm (confirmed)
- Henning Stahlberg, C-CINA, Basel (confirmed)
- Iban Ubarretxena-Belandia, CUNY, NY (confirmed)
- Janet Vonck, MPI, Frankfurt, Germany (invited)
- Juergen Plitzko, MPI, Martinsried, Germany (invited)
- Marcel Arheit, C-CINA, Basel (confirmed)
- Michael Landsberg, Univ. Queensland, Brisbane (confirmed)
- Nicolas Coudray, Univ. Mulouhse, France (confirmed)
- Nobuhiko Gyobu, AIST, Tokyo (confirmed)
- Per Bullough, Univ. Sheffield, UK (confirmed)
- Rafael Abela, SwissFEL, PSI, Villigen (confirmed)
- Rasmus Schroeder, Uni Frankfurt, Germany (invited)
- Richard Henderson, MRC, Cambridge, UK (confirmed)
- Richard Hite, Harvard Med. School, Boston (confirmed)
- Tom Walz, Harvard Med. School, Boston (confirmed)
- Torsten Schwede, Biozentrum, Basel (confirmed)
- Werner Kühlbrandt, MPI Frankfurt (confirmed)
- Xiangyan Zeng, FVSU, Gorgia, USA (confirmed)

Dates
Workshop Lectures: August 2-6, 2010 (Monday-Friday) Arrival: Sunday, August 1, 2010. Departure: Saturday, August 7, 2010

Program
The program is available at
http://www.2dx.unibas.ch/workshop/2010

Workshop Organizer
Karen Bergmann, University of Basel, Switzerland. Phone: +41 61 387 32 31. Email: Karen.Bergmann-at-unibas.ch

Scientific Organizers
Thomas Walz, Harvard Medical School, Boston, MA, USA.
Andreas Engel, University of Basel, Switzerland.
Henning Stahlberg, University of Basel, Switzerland.

Location
C-CINA (Centre for Cellular Imaging and Nano Analytics)
Biozentrum, University Basel
WRO-1058
Mattenstrasse 26
CH-4058 Basel
Switzerland
A description how to find the laboratory can be found at
http://tinyurl.com/cina-basel.


Registration
The workshop is currently limited to 30 participants. The fee for academic participants is CHF 300.-, which covers registration, breakfast and lunch during the workshop. Travel and Lodging are not included in the registration fee.
Registration is open now until June 15, 2010, and is available online at
http://www.2dx.unibas.ch/workshop/2010/workshop-registration

Festivals Before Workshop
If you like to spend some days in Switzerland before the workshop starts, you could
- enjoy the wonderful nature of Switzerland or the historic city of Basel,
- look at the BlueBalls festival in Luzern, one hour by train from Basel, see http://www.blueballs.ch/
- or look at the Stimmen festival in Loerrach, Germany, just accross the border a few minutes by public transport from Basel, see http://www.stimmen.com/
- July 31, the night before the workshop, is the Swiss National Festival with a big firework at 22:40, see http://www.basel.ch/programm_bundesfeier_2009.pdf
That should get you over the jetlag....

We hope to see you here in August,

Tom, Andreas, Karen, and Henning.


___________________________________________________

Henning Stahlberg
Center for Cellular Imaging and Nano Analytics (C-CINA)
at the Department of Biosystems Science and Engineering (D-BSSE)
Structural Biology and Biophysics, Biozentrum,
WRO-1058, Mattenstrasse 26
University Basel, CH-4058 Basel, Switzerland
Tel: +41-61-387 32 62 (office)
Tel: +41-61-387 32 31 (Karen Bergmann, administrative assistant)
Fax: +41-61-387 39 86
mailto:Henning.Stahlberg-at-unibas.ch
Skype:henningstahlberg
http://c-cina.org
http://2dx.org
___________________________________________________







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25, 27 -- 2010, in Basel, Switzerland
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From scott.lindsey-at-steton.com Sat May 22 09:29:03 2010
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Has anybody stored / archived (on the harddisk or somewhere else) the following
article in scanned / digital format?

Umrath W: Berechnung von Gefriertrocknungszeiten fur elektronenmikroskopische
Präparation. Mikroskopie 40, 9 (1983)

If possible, please send it to me - this is much appreciated.
Thank you in advance, and kind regards,
Reinhard


--

PD Dr. Reinhard Rachel
Universitaet Regensburg
Centre for EM - NWF III -
-at-Institute for Anatomy
Universitaetsstr. 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29

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Position Opening Sales Specialist - Western US
(Territory of AK, HI, WA, OR, CA, NV, ID, UT, AZ, MT, WY, CO, NM, TX)

This position will provide direct sales interaction with Bitplane's US
potential customer base for its complete range of software products.
Candidate will have the opportunity to develop the market in their
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and increased earning potential. Position is ideal for candidates that enjoy
travel but can also work easily out of their home office in their region.

The requirements for this position are:

Confocal, widefield, live-cell and/or digital imaging experience
Experience with microscopy image data processing
Outgoing personality with an affinity for self-motivation and responsibility

Excellent communication skills, oral, and written
Ability to vertically interface with multiple levels of diverse professional
clients

Desirable, but not required experience:

Technical knowledge of PC hardware, subsystems, peripheral and networking
equipment
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The duties of this position include:

Customer visits and analysis of customer's imaging needs.
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At least 50% travel will be required. Representative is required to live in
the territory they cover. Benefits include a base salary, performance based
commission, 401K plan, healthcare, and vacation. Representative will work
out of a home office.

Please email CV and cover letter to:

Michael C. Wussow
Vice President and General Manager Bitplane Inc.
mike-at-bitplane.com


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Email: bob.price-at-uscmed.sc.edu
Name: Bob Price

Organization: Univ South Carolina School Medicine

Title-Subject: [Filtered] Basic Confocal Microscopy Workshop

Message: A few slots remain in the Basic Confocal Microscopy Workshop
which will be held from June 14-18, 2010 at the University of South
Carolina. This is a well established workshop having been offered for
the past several years. Details are below:


The University of South Carolina School of Medicine Instrumentation
Resource Facility will be hosting the 6th Annual Workshop on Basic
Confocal Microscopy from June 14th through the 18th.

The workshop is directed towards beginning and intermediate users of
confocal microscopes and involves a series of lectures (specimen
preparation, labeling strategies, proper set-up of instrument
operating parameters, proper handling of 2D and 3D confocal images in
programs such as Adobe Photoshop and AMIRA), hands on specimen
preparation, and time on a number of different point scanning and
spinning disk confocal microscopes.

Companies scheduled to have instruments and applications experts
on-site include BioVision Technologies, Leica, Nikon, Olympus, Perkin
Elmer and Zeiss.

Faculty will include Dr. Jay Jerome of Vanderbilt University, Dr.
Ralph Albrecht of the University of Wisconsin Madison, Dr. John
Mackenzie of North Carolina State University, Dr. Tom Trusk of the
Medical University of Wisconsin, and Dr. Bob Price of the University
of South Carolina.

For further information please see:
http://dba.med.sc.edu/price/irf/irf.htm or contact Anna McNeal
(anna.mcneal-at-uscmed.sc.edu) or Bob Price (bob.price-at-uscmed.sc.edu).


Bob Price
Research Professor
USC School of Medicine
6439 Garner's Ferry Road
Columbia, SC 29209
803-733-3392 (T)
803-733-3212 (F)


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From: Wadowska-at-upei.ca
Date: Mon, 24 May 2010 15:00:18 -0500
Subject: [Microscopy] viaWWW: TEM turbo molecular pump

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Email: Wadowska-at-upei.ca
Name: Dorota Wadowska

Organization: Atlantic Veterinay College at UPEI

Title-Subject: [Filtered] TEM turbo molecular pump

Message: Hello,
Recently we installed a new turbo molecular pump (Edwards STP-301) on
our H7000. I noticed that the speed fluctuates a bit (1%). Is this
something that I should be concerned about or is this acceptable?
TIA
Dorota

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From: yi.liu-at-oregonstate.edu
Date: Mon, 24 May 2010 15:01:10 -0500
Subject: [Microscopy] viaWWW: Postdoctoral Position Available ot Oregon State Univ

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Email: yi.liu-at-oregonstate.edu
Name: Yi Liu

Organization: Oregon State University

Title-Subject: [Filtered] Postdoctoral Position Available ot Oregon State Univ,

Message: Colleagues,
I am sending this message for a professor at
Oregon State University. There is a postdoctoral
research position available immediately. Please
see the details below:

Thanks for your attention.

Postdoctoral Research Associate: Oregon State
Universityís Mid-Columbia Agricultural Research
and Extension Center announces the availability
of a 1.0 FTE 12-month fixed term position that is
part of a collaborative effort with Washington
State University. Requires Ph.D. degree in
horticulture, plant physiology or plant
science-related field. Focus of research will be
to characterize cell division and expansion
cycles throughout sweet cherry fruit ontogeny
through the development and application of
histological techniques, including confocal
microscopy, flow cytometry, and scanning electron
microscopy. The role of plant growth regulators
and modified cropload on cherry fruit size will
also be investigated. The position is located in
the beautiful Hood River Valley. The region is
renowned for its world-class windsurfing,
mountaineering, skiing and snowboarding, and
scenic hikes. Regarded for its local wineries,
brewpubs, and artisan shops, Hood River has a
population of approximately 5,000 people, and
offers quick access to the urban amenities of
Portland (65 miles). To review full posting and
apply, visit http://oregonstate.edu/jobs, posting
#0005618. For full consideration apply on or
before 05/30/10. OSU is an AA/EOE


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From: gary-at-gaugler.com
Date: Mon, 24 May 2010 15:35:38 -0500
Subject: [Microscopy] Re: Film and digital imaging

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

This topic has come up before. Useful reading about film
is Ansel Adams: The Negative. Using his Zone system, the
traditional S-shaped MTF curve of film can be linearized
from below the bottom axis and below the top axis. Digital
can be linear until their bins fill up and are just empty
of content due to saturation.

Fine art film work is/was not easy as one had to experiment
with each emulsion/brand type to maximize the linear region
for each. A densitometer was essential for this.

The next reading is Adam's The Print. This adds another
dimension to the film process. With digital, it is imperative
to evaluate a scene's histogram and be sure that there is
detail in the highlights as well as in the shadows. This is
where the Zone system excelled...especially on E-6. But regular
b/w were the preferred media for fine art pix.

Point and shoot digital cameras are more like point and guess or
point and hope for the best. The higher end cameras display
histograms (RGB, or you can throw color away).

Snappy b/w can be done without the Zone System if exposure is
correct and mates to paper grade. The Zone System produces
more subtle shades of grey with a soft feel--probably not what
you seek.

gary g.



At 02:50 PM 5/21/2010, you wrote:



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From: tbargar-at-unmc.edu
Date: Mon, 24 May 2010 16:24:01 -0500
Subject: [Microscopy] LR White and LR Gold comparisons

Contents Retrieved from Microscopy Listserver Archives
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Can anyone give me a comparison of LR White and LR Gold?


Tom Bargar
University of Nebraska Medical Center
Core Electron Microscopy Research Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

==============================Original Headers==============================
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From: PhillipsT-at-missouri.edu
Date: Mon, 24 May 2010 16:27:48 -0500
Subject: [Microscopy] LR White and LR Gold comparisons

Contents Retrieved from Microscopy Listserver Archives
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It depends on your application. LRG is UV polymerized, LRW is generally polymerized with heat. LRW works with osmicated tissue, LRG doesn't. It is a lot easier to get nice thin sections of LRG compared to UV polymerized Lowicryls. I like LRG for EM immunocytochemistry but if you want to do immuno-staining of 0.5 um sections, nothing compares to BMMA.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: tbargar-at-unmc.edu [mailto:tbargar-at-unmc.edu]
Sent: Monday, May 24, 2010 4:25 PM
To: Phillips, Thomas E.

Can anyone give me a comparison of LR White and LR Gold?


Tom Bargar
University of Nebraska Medical Center
Core Electron Microscopy Research Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

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From: tbargar-at-unmc.edu
Date: Mon, 24 May 2010 16:33:35 -0500
Subject: [Microscopy] TEM protocol for enhancing membranes

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

I have a PostDoc who would like to see the membranes of the vesicles she
is interested in "stand out" better from the cytoplasm in the cells. She
brought me an old paper where the fixation protocol pretty much extracted
everything out the cells, except membranes. Similar to the days of using
Potassium Permaganate, etc. I would appreciate any and all advice on
fixation protocols that may help to enhance the visibility of membranes,
while maintaining a good fixation of the cells without extraction of the
cytoplasm, etc.


Tom Bargar
University of Nebraska Medical Center
Core Electron Microscopy Research Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

==============================Original Headers==============================
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From: PhillipsT-at-missouri.edu
Date: Mon, 24 May 2010 16:43:20 -0500
Subject: [Microscopy] TEM protocol for enhancing membranes

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

I have had good success with the OTO method - osmium thiocarbohydrazide osmium of Seligman et al. 1966. You won't believe how black your tissues turn! See Willingham and Rutherford 1984 J Histochem Cytochem 32:455-460 http://www.jhc.org/cgi/reprint/32/4/455.pdf for a good start.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: tbargar-at-unmc.edu [mailto:tbargar-at-unmc.edu]
Sent: Monday, May 24, 2010 4:34 PM
To: Phillips, Thomas E.

I have a PostDoc who would like to see the membranes of the vesicles she
is interested in "stand out" better from the cytoplasm in the cells. She
brought me an old paper where the fixation protocol pretty much extracted
everything out the cells, except membranes. Similar to the days of using
Potassium Permaganate, etc. I would appreciate any and all advice on
fixation protocols that may help to enhance the visibility of membranes,
while maintaining a good fixation of the cells without extraction of the
cytoplasm, etc.


Tom Bargar
University of Nebraska Medical Center
Core Electron Microscopy Research Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

==============================Original Headers==============================
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15, 37 -- From PhillipsT-at-missouri.edu Mon May 24 16:43:20 2010
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15, 37 -- From: "Phillips, Thomas E." {PhillipsT-at-missouri.edu}
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15, 37 -- "microscopy-at-microscopy.com"
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From: rosemary.white-at-csiro.au
Date: Mon, 24 May 2010 17:24:55 -0500
Subject: [Microscopy] TEM protocol for enhancing membranes

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

We use reduced osmium as a secondary fixative for membranes. Works
especially well on cultured cells.

Pat Kysar
University of California, Davis
Medical School, Pathology
EM Lab

----- Original Message -----
X-from: {PhillipsT-at-missouri.edu}
To: {pekysar-at-ucdavis.edu}
Sent: Monday, May 24, 2010 2:49 PM

I don't know if this works for animal tissues, or works as well as the OTO
method, but a less toxic (yet equally tissue-blackening) protocol involves
incorporating tannic acid into glutaraldehyde and/or formaldehyde fixative,
then "developing" with ferric chloride. Alternatively, just adding tannic
acid to the fixative before osmium staining can also enhance membranes.

For an older review: Chaplin, AJ (1985) Tannic acid in histology: an
historical perspective. Stain Technology 60: 219-231

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au


On 25/05/10 7:47 AM, "PhillipsT-at-missouri.edu" {PhillipsT-at-missouri.edu}
wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} I have had good success with the OTO method - osmium thiocarbohydrazide osmium
} of Seligman et al. 1966. You won't believe how black your tissues turn! See
} Willingham and Rutherford 1984 J Histochem Cytochem 32:455-460
} http://www.jhc.org/cgi/reprint/32/4/455.pdf for a good start.
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://www.biology.missouri.edu/faculty/phillips.html
} http://www.biotech.missouri.edu/mcc/
}
}
} -----Original Message-----
} X-from: tbargar-at-unmc.edu [mailto:tbargar-at-unmc.edu]
} Sent: Monday, May 24, 2010 4:34 PM
} To: Phillips, Thomas E.
} Subject: [Microscopy] TEM protocol for enhancing membranes
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} I have a PostDoc who would like to see the membranes of the vesicles she
} is interested in "stand out" better from the cytoplasm in the cells. She
} brought me an old paper where the fixation protocol pretty much extracted
} everything out the cells, except membranes. Similar to the days of using
} Potassium Permaganate, etc. I would appreciate any and all advice on
} fixation protocols that may help to enhance the visibility of membranes,
} while maintaining a good fixation of the cells without extraction of the
} cytoplasm, etc.
}
}
} Tom Bargar
} University of Nebraska Medical Center
} Core Electron Microscopy Research Facility
} 986395 Nebraska Medical Center
} Omaha, NE 68198-6395
} 402-559-7347
} tbargar-at-unmc.edu
}
} ==============================Original Headers==============================
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10, 40 -- Subject: Re: [Microscopy] RE: TEM protocol for enhancing membranes
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From: s.h.coetzee-at-gmail.com
Date: Tue, 25 May 2010 03:13:04 -0500
Subject: [Microscopy] Film and digital imaging

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

I agree tannic acid is a good approach. It is important to use the "right" type of tannic acid. I think it is the low molecular weight species but I would check this before proceeding. Tom


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: rosemary.white-at-csiro.au [mailto:rosemary.white-at-csiro.au]
Sent: Monday, May 24, 2010 5:26 PM
To: Phillips, Thomas E.

I don't know if this works for animal tissues, or works as well as the OTO
method, but a less toxic (yet equally tissue-blackening) protocol involves
incorporating tannic acid into glutaraldehyde and/or formaldehyde fixative,
then "developing" with ferric chloride. Alternatively, just adding tannic
acid to the fixative before osmium staining can also enhance membranes.

For an older review: Chaplin, AJ (1985) Tannic acid in histology: an
historical perspective. Stain Technology 60: 219-231

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au


On 25/05/10 7:47 AM, "PhillipsT-at-missouri.edu" {PhillipsT-at-missouri.edu}
wrote:

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}
} I have had good success with the OTO method - osmium thiocarbohydrazide osmium
} of Seligman et al. 1966. You won't believe how black your tissues turn! See
} Willingham and Rutherford 1984 J Histochem Cytochem 32:455-460
} http://www.jhc.org/cgi/reprint/32/4/455.pdf for a good start.
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://www.biology.missouri.edu/faculty/phillips.html
} http://www.biotech.missouri.edu/mcc/
}
}
} -----Original Message-----
} X-from: tbargar-at-unmc.edu [mailto:tbargar-at-unmc.edu]
} Sent: Monday, May 24, 2010 4:34 PM
} To: Phillips, Thomas E.
} Subject: [Microscopy] TEM protocol for enhancing membranes
}
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} I have a PostDoc who would like to see the membranes of the vesicles she
} is interested in "stand out" better from the cytoplasm in the cells. She
} brought me an old paper where the fixation protocol pretty much extracted
} everything out the cells, except membranes. Similar to the days of using
} Potassium Permaganate, etc. I would appreciate any and all advice on
} fixation protocols that may help to enhance the visibility of membranes,
} while maintaining a good fixation of the cells without extraction of the
} cytoplasm, etc.
}
}
} Tom Bargar
} University of Nebraska Medical Center
} Core Electron Microscopy Research Facility
} 986395 Nebraska Medical Center
} Omaha, NE 68198-6395
} 402-559-7347
} tbargar-at-unmc.edu
}
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From rex-at-vanwertinc.com Mon May 24 18:19:28 2010
Return-Path: {rex-at-vanwertinc.com}
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I am not a technical guru on film or digital. But have been in EM for
many years. Film is expensive and in the old days your budget limited
the amount of images you could took. Digital no extra cost for more
images. Often the image you took was not perfect and the chance of
finding that exact area often was nearly impossible unless you did
draw maps by hand of your sample as you explored it. (yes that was how
we did it long time ago..) Especially TEM, your plates are with other
peoples images and development was only once once the cassette is
full, a waiting time before you could see your outcome. Then the sad
part was to phone occasionally the user with the news you did
something wrong with the chemicals and the images are blank..., under
or totally over developed. The time fund and energy lost in those odd
times left emotional imprints. I never want to go back to film.
For dissertations all the prints we made, and corrections often meant
reprints. Gluing on of Micronbars, letter transfer from “scratch off
sheets” Cepachrome negatives for slides, a lot of work for conference
presentations. In the early days of PowerPoint, you have to put in a
large amount of work to get your images into PowerPoint.
Nostalgic about film, but converted.


On Mon, May 24, 2010 at 10:41 PM, {gary-at-gaugler.com} wrote:
}
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} This topic has come up before.  Useful reading about film
} is Ansel Adams: The Negative.  Using his Zone system, the
} traditional S-shaped MTF curve of film can be linearized
} from below the bottom axis and below the top axis.  Digital
} can be linear until their bins fill up and are just empty
} of content due to saturation.
}
} Fine art film work is/was not easy as one had to experiment
} with each emulsion/brand type to maximize the linear region
} for each.  A densitometer was essential for this.
}
} The next reading is Adam's The Print.  This adds another
} dimension to the film process.  With digital, it is imperative
} to evaluate a scene's histogram and be sure that there is
} detail in the highlights as well as in the shadows.  This is
} where the Zone system excelled...especially on E-6.  But regular
} b/w were the preferred media for fine art pix.
}
} Point and shoot digital cameras are more like point and guess or
} point and hope for the best.  The higher end cameras display
} histograms (RGB, or you can throw color away).
}
} Snappy b/w can be done without the Zone System if exposure is
} correct and mates to paper grade.  The Zone System produces
} more subtle shades of grey with a soft feel--probably not what
} you seek.
}
} gary g.
}
}
}
} At 02:50 PM 5/21/2010, you wrote:
}
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--
Stephan H Coetzee
EM Scientist
Electron Microscope Unit
University of Botswana


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From: oshel1pe-at-cmich.edu
Date: Tue, 25 May 2010 10:06:12 -0500
Subject: [Microscopy] RE: TEM protocol for enhancing membranes

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

Hi John et al,

Thanks for the interesting quantifying TEM 'film scanning' post and the recent posts. My post is a bit off-target as I have no requirement to scan largish format Kodak 4489 TEM B&W film these days, just 120 B&W film and 35mm colour slide/negatives [and being an optical microscopist unlike the EM guys I do dream in colour]. Looking at my film scans of semi-pro colour slide film used for photos in the 1970s and 1980s, it is clear that my modern digital SLR camera is capturing more detail than my old film SLR camera was thirty-five years ago. However this is partly because I compare the digital photo with a digital scan of my slide [plus there's improved camera optics, sophisticated onboard digital image processing and the age of the film]. Essentially re-photographing the 'photograph' by scanning film always lost some ultrafine detail [although nowhere near as much as the original photo did of the view it was capturing]. Not that you would notice this difference much up to A4 print sizes, where the digitized scan and original colour slide look pretty similar.

Our Epson V700 scans tissue pathology sections on glass slides at the same resolution as our large research Nikon Eclipse TE2000U photo-microscope captures photos using a 1x objective & 5Mp colour camera [total optical magnification 10x - although that 5MP camera will be capturing the optical view in lower detail than actually present and the Nikon microscope scoots ahead resolution wise if you up the mag to a 4x objective]. I guess many modern EM digital cameras are capturing at a significantly lower resolution than the view as well [rather like our expensive low-noise optical microscope 1.3MP 1344x1024 fluorescence B&W cameras], and many EM digital cameras capture at theoretically lower resolution than traditional ultra-fine grain TEM B&W film. Kodak 4489 B&W film has a resolution reputedly significantly higher, perhaps more than 2x the 4000ish 'dpi' of colour slide film, although the cheap Epson V750 flatbed scanners do seem to recover a lot of detail from this Kodak film - Kodak just say 'ultra-fine grain' size for 4489]. A Gatan Ultrascan 4k × 4k CCD model US4000 has pixels of 15 um whereas Agfa 10E56 holographic film has a resolution of nearer 4,000 lines/mm — equivalent to a pixel size of 0.125 micrometres and apparently an active dynamic range of over five orders of magnitude in brightness, compared to typical scientific CCDs. This bodes well for Kodak 4489 compared to a CCD detector in terms of theoretical resolution, which is no doubt why film use has lingered on in niche applications. However lost detail of say a ship or face is easier for us to judge than more abstract images of cell ultrastucture. Plus film is more of pain to handle, easy to expose incorrectly [and you don't discover that until much later], non-reusable, difficult to calibrate and expensive per shot compared to digital capture [although the last quote I had, a few years ago now, for a Gatan 4kx4k TEM digital camera upgrade from film was £40,000+]. I now take 10,000+ digital photos a year compared to a few hundred back in!
the 197
0s, so the number of 'great shots' I get per year has increased significantly and I can now easily create stitched panoramas from multiple shots. Plus film, like the audio cassette and LP before it, is also doomed as manufacturer's simply lose interest in producing it for a contracting market.

Going back to scanning colour film, cheaper flatbed scans like the Epson V750 always seem to have that slightly 'soft-focus' image detail at maximum mag, although this can be improved slightly with light use of Photoshop's Unsharp Mask sharpen tool - I guess you never really know what any scanner is doing [it may be secretly applying subtle image processing algorithms, just as the digital SLR camera does]. Having grown up with 1950s 'red shirt' National Geographic prints, colour correction on a modern PC VDU seems hardly worth the trouble [and considerable expense] - I can set my camera colour tones to Vivid, Cool, 'Normal', etc anyway so it's almost adjust to taste - and I print so few photos outside of the Xmas card inserts these days I send those off to photo-labs so colour correction for my printer is irrelevant. I no longer bother with an expensive (£400) pro multi-grey/black/colour ink photo printer and just use a large HP Office Inkjet which is better than the office Laserjet for images and can still use photo-paper. Being an optical microscopist, even my B&W images are colourised by LUTs to RGB. I guess it is important to set VDU contrast/brightness to ensure you can actually see detail in shadows though [I have accidently reset the monitor to 'default' and not realized for a while that I could no longer see any detail in significant areas of the photo], and generally at work I avoid the budget 'multimedia' twisted-nematic (TN) LCD displays optimized for fast gaming/video [2 to 6 ms response] monitors and go for an IPS type 'professional grade' display [typical 14ms response time]. All my works images start off in the acquisition software and many eventually pass through Photoshop CS5 [I couldn't imagine life without Photoshop CS4/5].

X-from simply looking at the results of a [£10,000+] Hamamatsu Flextight 848 scan of decent prosumer 100 ASA colour slide film [Agfa/Perutz/Kodak] from the 1970s and comparing them with the likes of a 'budget' Epson V700/Canon 9950F scan, the only thing I really noticed was that the film grain seemed in sharper focus with the Flextight 848. It certainly looked like film grain and not some optical artifact, you can see similar comparison images in figure 5, p28:

Scanning film on a budget: http://www.well.ox.ac.uk/_asset/file/infocus-scanning-film-on-a-budget.pdf

Although the budget scanner used was the slightly 'inferior' Canon 9950F scanner, rather than an Epson V750 Pro. Being a photographer I naturally quantified the images by saying to myself 'It looks alright I suppose'. The actual detail within the scanned image wasn't really any different between the scanners, suggesting both scanners were pretty much capturing at or beyond 35mm colour slide film resolution - with the £250 Canon 9950F therefore scanning at a much cheaper [albeit much slower] rate. I also noticed that the original colour slide definitely had more detail than both the slide scans though, if you look at the slide film using a light box and an 8x inspection magnifier - and so see what exactly the scan really should look like [I suppose this view can thus be used as a kind of subjective* 'target' of how 'near' the scan of the film got to the original slide - although not to the original view photographed].

I guess one always wants to recover far more detail from film [and digital images] and mostly that detail simply isn't there, as it got lost when you captured the image in the first place [film and their scans always disappoints at high enlargement - and now this disappointment takes just a few mouse zoom clicks with a digitized image on a PC]. I guess I now wish my father had bought me a Hasselblad 70mm format film camera for my 9th birthday and not a Kodak 104 '126 format' Instamatic - although I soon abandoned that and took over his 35mm SLR.

Sadly I never got to play directly with the Flextight 848 [semi-drum] scanner myself, it was run within a university services by operators that perhaps were naturally more concerned with throughput than ultimate scan quality per slide. Fortunately with any scan of higher resolution B&W TEM film you always have the original negative [that has an estimated archive life of 500 years, far than the digital files produced from it - I doubt if a large digital SEM image file would survive more than 30 years without corruption unless archived very carefully, and I'm probably being optimistic]. Mind you most film, particularly negatives [or details of who/what they were images of], ends up lost in time as well during lab and loft clearances - with even the BBC having to chase after the dustcart on numerous occasions.

Regards

Keith

*Or as Woody Allen put it 'Yes, but objectivity IS subjective'.

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford OX3 7BN,
United Kingdom.

Telephone: +44 (0)1865 287568
Email: kjmorris-at-well.ox.ac.uk
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy

-----Original Message-----
X-from: john_mackenzie-at-ncsu.edu [mailto:john_mackenzie-at-ncsu.edu]
Sent: 15 March 2010 18:52
To: kjmorris-at-well.ox.ac.uk

Folks:

This question has come up before and I have additional data to add.
Let me preface first by stating that my laboratory is not involved in
rigorous testing of images for three dimensional image reconstruction
where I personally believe another level of precision is achieved/required.
I was curious because all of my measurements of the the Epson
4880/4990/D750M, D750M pro indicated that Epson produced images of TEM
negatives scans that were CLEARLY superior to everybody elses (including
older Epson scanners. This was based on examining details of TEM and
comparing them under a microscope with actual photographs of the same
negative printed in a darkroom on a Durst enlarger with Agfa Brovira
film. (For you younger guys film is the stuff we used to use in the dark
ages to record images. The darkroom equipment was "the best"). From this
point forward dots per inch which is a meaningless term, will be
replaced exclusively by pixels per inch.

Although careful one to one examination showed that every detail was
retained, indicating the grayscale resolution and spatial resolution was
sufficient), there was no doubt that a trained microscopist could pick
out the darkroom printed images every time. "They were sharper" (This is
not a quantifiable entity. Before you jump all over me read on)
I decided to "calibrate" my scanner. As noted in the previous post, one
the best measures of the resolution of the system is the MTF. To make an
eight month quest short, Don Wilson at Kodak would wrote the ISO
standard on scanner resolution helped me measure the MTF. It was
difficult to believe that a flatbed scanner that cost $800 was beating
the resolution of drum scanners that cost $27,000- 100,000. These
scanners however had been rigorously tested by "disciples" from the MRC
and were known to produce excellent data.

Film "experts (photographers some of whom were engineers made logical
blunders and confused the MTF of the film with the MTF of the camera)(
see work of Ken Taylor's group on the resolution of film by diffraction
criteria) The "best" photographers argue whether the resolution of film
is 4000 pixels per inch or 8000 pixels per inch.

There are clear answers to this issue that I should be writing up for
Microscopy Today but are not necessary to understand right now. The
important point is that consensus is greater than 4000 pixels per inch.
(Don't bother with your arguments about microns/pixel, etc. I know, I
know .. not now)

So I went out and purchased $3,400 worth of targets and "standards" and
measured the scanners MTF. I thought I would a hard quantifiable number
that was "unbiased" by human variability. Nope.

The MTF requires the user to define the noise limit that is "acceptable"
for discerning gray levels. Photographers have a range that is pretty
well defined because images that fall below a certain contrast level
"look" too grainy and therefore are clearly "unacceptable" BUT a
scientist/microscopist looking at that image sees that it is noisy but
wouldn't reject it until the niose level was two, three, or four times
worse (in rare cases, even worse than that ) Additionally the MTF is
dependent on the size of the scanned pixel and there are real issues
with the number of grayscale values required and the size of the scanned
pixel. (This is a question of whether we are looking at the resolution
of the film as defined by it's ability to resolve diffraction data or 12
bit gray scale data. I think a films spatial resolution is higher when
defined by diffraction critera than by 12 bit imaging criteria. (This
opens yet another can of worms)

Finally, (I love this one) the MTF measures contrast and frequency. It
does not and cannot measure sharpness. whoops. What measures sharpness.
not edges cause the MTF measures edges to get contrast and frequency.
The photographers use that ultra precise measuring tool called the
photographer and he/she says, " Well it looks sharper" The examples
shown in the majority of cases would never pass scientific scrutiny.
My tests were based on the assumption that I needed more than eight bits
so I chose twelve bits per pixel as my necessary lower limit.
Using a target for scanner reflective image resolution measurement based
on contrast versus resolvable lines per inch (which can be
mathematically converted to pixels per inch.

The target showed that all the scanners listed above could achieve 2000
pixels per inch resolution. This is the upper limit of all the targets
including the USAF target and the ISO target that is commercially available.
I had two 4990's that were measured and surprisingly one passed one did
not. The one that didn't was far worse in one axis versus the other. but
both failed.

Working with Don Wilson at Kodak (who by the way authored ALL the iso
specs), we measured all at a little better than 2000 pixels per inch
when using a noise level I figured was acceptable to most microscopists.
His measurements showed clearly and quantifiabley that one 4990 was broken
Those of you who are really paying attention will note that I said
reflective and we are more concerned about transparency scanning. In the
Epsons transparency scanning is not the same as reflective scanning. So
one should test transparency scanning in the same manner. Good luck.
There are NO targets available. Seems there is ZERO demand. I really
mean ZERO. I had some long discussions with the manager at the company
the makes the reflective target and he agreed to make me a custom target
for transparency. (I understood that it might have a minor problem with
certification as a "standard" but I knew that it could tell me if it
coulds see 2000 pixel per inch in transparency mode in both directions
and that that would work for me.

Got the target and all passed but the broken one. Same with the MTF all
passed but the broken one.

Here is my take on the data above.

Specifications listed by the manufacturers are total bull, have no
scientific validity, do not pass any scientific test and are all subject
to human interpretation. (It is that noise ratio thing that I can't get
nailed down for TEM images as we would normally view them in
conventional stained embedded TEM.)

The MTF does not measure sharpness which as you might recall was the
clear difference between the dark room printed image and the scanned
and printed image.

Let me throw in another line of experimentation we did. We wanted to
know if an eight megapixel digital camera printing images at 19 inches
by 13 inches captured and printed images that were limited by the
resolution of either the camera's detector or by the printers
resolution. (Although there is an iso specification on scanning
resolution there has never been one done on printers. Three years ago
when I last spoke with Don he as working on it but he was concerned that
Kodak may not want to continue with that line of experimentation
(externally))

The answer was that we had to more than half the resolution before we
could even detect the effects of a sharpening filter without a microscope.
The final data that is available now comes from the photographic
experts. They don't agree on anything except. The really good people
believe that film can resolve 4000 pixels per inch OR 8000 pixels per
inch if scanned in a high quality drum scanner under liquid immersion.
(NEVER a flat bed)
My take on discussions with groups that scanned film and then used the
data for critical reconstructions was that they were closer to the 4000
to 8000 pixel per inch number. Ken Taylor group has I believe an even
higher number because the spots can achieve a higher resolution than a
12 bit grayscale image pixel. (I'm not going to debate this because it
isn't relevant to the topic here)

I think a vast majority of the experts that were using film for
reconstruction and were active in the measurement of it's limitations
are now using digital acquisition systems that increase their
productivity more than 10 fold over working manually. Because everything
can be automated, the productivity can skyrocket beyond that.. The
scientific fact that the detector can measure all the detail available
has been determined critically. However, because film has a much higher
resolution the field of view is decreased substantially. This fact is
less than irrelevent if you are collecting data at ten, a hundred or
possibly even more than you could prior to digital automation. The
reduction in field of view can be a problem for say a pathologist. The
reconstruction groups love that the detectors are linear. As an imaging
person I can tell you that this fact is the biggest PROBLEM with digital
systems but that's another major tome.

Pulling it together for the big climax.

When we printed negatives in the darkroom we could blow up the images
many fold. A 35 mm film can easily be blown up to 13 x 19. which is
greater than ten. Thats a minimum. By the same logic our film can
therefore be blown up 10 fold to 40in x 52. (3.25 x 4 times 13) We only
print at a maximum of 8 x 10 and much less for the journals. (The 40 in
x 52 in format is quite rare.) So we don't use all that film has. and
under most conditions we can therefore drop the resolution we need from
the discussion of the upper limit at 4000 ppi or 8000 ppi to the 2000
ppi that the scanner can actually do when working properly and operated
properly. (Now I would say calibrated properly but that is problematic.
The target that I got for measuring the MTF of the transparency scanner
was a gift and was produced on a classified, propriety scanner that is
no longer available.Because the Epson scanners use the same scanning
mechanism for both reflective and transparency scanning you can at least
infer that things are right ( My data suggests that there is a strong
correlation of the two and I have never seen the transparency messed up
without the reflective being messed up also. Doesn't mean it can't
happen but it is unlikely.

We are 99% convinced that the film has at least 12 bits of data. We scan
at 16, store at 16 and then convert to 8 for most imaging. Again another
tome but this has no controversy among the groups that have really
looked at the data.

We do nothing but scan negatives for all our digital. We scan routinely
at 1200 ppi, 16 bit grayscale full size and store as tif 16 bit. This
gets us most of the data that 8 x 10 would get us from the past. When we
want high resolution we are normally after a smaller selected area and
can afford to scan at the higher resolution.

The Epson scanners are the only scanners I even consider. Measurements
show that they get at least 2000 ppi and although that is not comparable
to film it is equivalent to a 4k by 4k digital camea. (do a price
comparison and the scanner has a slight advantage). Most TEMs in the
future will take most of their images by digital and less by film. Soon
we will go the same route that the fashion, ad, and high end
professional photographers and the detector will hit film resolution
and then go beyond it. ( Hey, it's only sand and technology after all.)
The price will fall and guys in the future will wonder what the heck
those people from the past were debating.(And what is that film stuff
anyway?)

As I am about to climb on my soapbox, dear readers who are satisfied can
stop reading.

One of the major problems that I see going forward is that students
actually think they know what they are doing. From the standpoint of
understanding digital imaging sufficiently to make critical ethical and
scientific decisions on what they can do and what they can't they are
positively clueless. Clueless people who think they know what they are
doing are bond to make serious mistakes. Is there increased interest in
properly educating them in the science of imaging? No Do students and
advisors think that students need this training ? No

With the increased use in digital data manipulation, how often are
programs tested for scientific validity? Do you know the answer to that
question? Are you using Photoshop? Do you understand that there is an
issue with how Adobe handles 16 bit numbers?

We use transparency scanners and yet no target is commercially available
to measure MTF (Please notify the list if you know a source)
I think that we actually need more data and more serious discussion as I
see it nearly impossible to clearly define what is ethic to do to an
image and what is unethical if you can't define the original data with
any degree of precision.

I stopped Chairing Symposia because too many individuals complained that
the topic had been beaten to death and that there was nothing new. I am
probably not going to give my Digital Imaging Course because students
and colleagues feel that the course is not necessary. I neither want to
Chair the Symposia nor teach the course but would ask that smeone
actually remember that we are doing science and that in science you need
controls and you need to understand how all the instruments that measure
things work. When you don't and you ASSUME you knoe how it works you are
headed for trouble.

Charlie Lyman cleverly put me on the Editorial Board of Microscopy Today
so the above should make at least a three part article in the coming
months . In addition, I am working on a handbook which includes much of
the practical workflow with scientific justification for the Sunday
Short Course. this year. (which will likely be my last) Now if I can
just stay healthy and alive long enough to complete thse tasks, I'll be
happy.

Sorry way too much soapbox but if you followed my directions you are not
reading this anyway.

Now, if this doesn't stir up a hornets nest of controversy, I will be
shocked. BUT please be advised that I didn't have enough time to write
this so that it is unlikely I'm going to get into a battle of verbal
jousting on the list server. Make sure that before you leap into this
you are clear about the MSA statement on ethical digital imaging. Since
the data is scientific data, there is little that you can do without
having to report your manipulations in detail. In addition, since it is
data, how you acquired that data must be understood and should in my
opinion be validated.

John Mackenzie

john_mackenzie-at-ncsu.edu

--

John Mackenzie, Jr. PhD

Coordinator for the Center for Electron Microscopy

Professor of Microbiology

North Carolina State University

Phone (919) 515-2664 Fax (919) 515-8293


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Email: charlesping-at-hotmail.com
Name: Xiaoping Zha

Organization: University of York, UK

Title-Subject: [Filtered] JEOL 6400F SEM circuit diagrams needed

Message: Deal all

We have a JEOL 6400F SEM moved to a new room and it needs baking for
the electron column. However, the baking controller cannot start the
process. We want to check the circuit connections but cant find our
circuit diagrams. Does anyone have a copy of the circuit digrams? It
would be perfect if it is in pdf file. Or if you are in UK, we can go
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Thanks

Dr. Xiaoping Zha
Department of Electronics
Univetrsity of York

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Tom is right: it's important to use the monomeric kind, like Mallinckrodt 1764.

Phil

} I agree tannic acid is a good approach. It is important to use the
} "right" type of tannic acid. I think it is the low molecular weight
} species but I would check this before proceeding. Tom
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
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From: underwoo-at-u.washington.edu
Date: Tue, 25 May 2010 10:32:43 -0500
Subject: [Microscopy] Re: Film and digital imaging

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Dear All,

This thread has been interesting in bringing up many of the changes that have occurred during the transition from film to digital and I feel comfortable with the details. But it makes me wonder about changes other than film to digital media that has affected the final visual impact of TEM images. One change, in particular, is the accelerating voltage of the scopes. I think starting out I was using 20kV and then it progressed to 40, 60, 80, 120kV until the newer scopes that I could access were are no longer usable for biologic specimens at } 200kV and as multi-user facilities it was not encouraged or convenient to reset the voltage down to 40kV and realign. Through this progression of higher energy I have watched an apparent reduction in contrast until I actually can’t find the sample. When still using film I compensated through exposure and development. Then with digital this became a Photoshop
adjustment but does anyone know if the inherent dynamic range of a biologic thin section, especially a lightly stained biologic cryothin section, is reduced as you use higher accelerating voltage to image? If so, this is a loss than can’t be recovered using film or digital sensors.

Robert Underwood
University of Washington
Dermatology







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From: frah0010-at-umn.edu
Date: Tue, 25 May 2010 19:18:23 -0500
Subject: [Microscopy] Co in olivines

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I have been enjoying the discussion about digital and film, but I want to
make comment here: Digital image acquisition is no silver bullet. You cannot
recover digitally what you don’t have physically. To have a good digital
image, one needs to start out with a good physical signal. While digital
imaging can help with difficult situations and make a small contrast more
visible, one has to have a contrast to begin with. What I want to say is
that digital imaging does not absolve the TEM user from knowing the
instrument and its limitations and capabilities.

A TEM observation or experiment involves many steps, from specimen
preparation to TEM calibration, Astigmatism correction, selecting the
appropriate acceleration voltage, to knowledge about electron sample
interaction, contrast mechanisms, to selecting the right imaging conditions
and knowing the imaging parameters, and for film, to choosing the right
paper. For the best results, all steps in this chain must be optimized,
which requires an in-depth knowledge of all these factors and steps.
 Digital imaging is only one step in this whole chain. Yes, it can be used
to widen the parameter space for reasonable images, for example, low
contrast can be overcome to some extent by digital manipulation, but again,
for best results, every step has to be optimized, not just the imaging.

To me, one of the exciting aspects of digital imaging is that it integrates
the imaging tightly with the instrument, enabling not just a replacement of
film, but allows for an extension of the TEM capabilities through feedback
and automation.

---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode at ResAltaTech.com
www.ResAltaTech.com






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From tmanners-at-reveries.com Tue May 25 15:18:25 2010
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Micro folks,

Does anyone know of published (or even unpublished) values for cobalt in San Carlos, AZ olivine? How about Harrat al Kishb, Saudi Arabia forsterite? Both concentrations should be in low three-digit ppm range. I'd like to know because I'm trying to check our accuracy at measuring Co in olivine at such a level.

Thanks,
Ellery

-----------------
Ellery Frahm
Senior Research Fellow, Department of Geology & Geophysics
Manager & Principal Analyst, Electron Microprobe Lab
Doctoral Candidate, Department of Anthropology
University of Minnesota - Twin Cities campus
Lab website: http://probelab.geo.umn.edu/
Personal: http://web.mac.com/elleryfrahm/








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From: Bplowman-at-pacific.edu
Date: Tue, 25 May 2010 19:44:05 -0500
Subject: [Microscopy] viaWWW: Tannic acid for stain enhancenment ( mordant) to lead

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Email: Bplowman-at-pacific.edu
Name: Barbara L. Plowman

Organization: University of the Pacific, Dugoni School of Dentistry

Title-Subject: [Filtered] Tannic acid for stain enhancenment (
mordant) to lead

Message: You can use tannic acid as a mordant to lead to enhance
membranes stain. Simonesque and Simonesque, Journal of Cell
Biology, Jan. 1976 is the reference. I don't know if you can still
get EM grade tannic acid from Mallinquot or not. That's what you
need. That's what the Simonesque's used. It will enhance membranes
beautifully. Your fixation should be a standard glutarladehyde fix.

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From: mikehern-at-nist.gov
Date: Tue, 25 May 2010 19:45:00 -0500
Subject: [Microscopy] viaWWW: TEM Technician Vacancy Announcement

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Email: mikehern-at-nist.gov
Name: Mike Hernandez

Organization: NIST

Title-Subject: [Filtered] TEM Technician Vacancy Announcement

Message: Physical Science Technician
Type of Appointment: FTP
Relocation Expenses are Authorized

Duties:
Incumbent will train users of the Center for
Nanoscale Science and Technologyís (CNSTís)
national shared-use facility, the CNST NanoFab,
in the operation of an analytical scanning
transmission electron microscope STEM (FEI Titan
80-300), including basic microscope operation and
the use of x-ray and electron energy-loss
spectroscopy techniques. Perform preventative
maintenance and fault diagnosis of the microscope
and associated instrumentation, including sample
preparation equipment.
Incumbent will act as a secondary operator and
user trainer of the NanoFab scanning electron
microscopes (SEMs) and focused ion beam (FIB)
systems.
Incumbent will perform preventative maintenance
and fault diagnosis of the CNSTís environmental
scanning transmission microscope (ESTEM) (FEI
Titan 80-300) and associated instrumentation,
including sample preparation equipment. Assist
in the operation of the ESTEM. Assist in the
operation and development of new instrumentation
associated with the ESTEM.
Incumbent maintains and optimizes the electrical,
thermal and mechanical stability of the
microscopesí operating environment and optimize
the microscopesí performance.

Basic Requirements:

Degree in physical sciences, life sciences, or
engineering that included 30 semester hours in
physics, electrical engineering or materials
science, supplemented by course work in
mathematics through differential and integral
calculus, and at least 6 semester hours of
physics.
OR
Combination of education and experience -- course
work equivalent to a major as shown above,
including at least 30 semester hours in physics,
electrical engineering or materials science,
supplemented by mathematics through differential
and integral calculus, and at least 6 semester
hours of physics, plus appropriate experience or
additional education.

In addition to the basic requirements, you must have:

3 years of progressively higher level graduate
education leading to a Masters degree or Ph.D.
degree or equivalent
OR
One year of specialized experience equivalent to
at least the ZT-II (GS-9 equivalent) level.

Specialized experience is defined as experience
in the operation and maintenance of transmission
electron microscopes, including experience with
low- and high-voltage power supplies, and
diagnosis and repair of vacuum and mechanical
systems

Applicants must meet requirements by the closing date of this announcement.

To apply go to USAJOBS.GOV Announcement number: NISTCNST-2010-0005


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From: Bplowman-at-pacific.edu
Date: Tue, 25 May 2010 19:45:31 -0500
Subject: [Microscopy] viaWWW: LRWhite

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Email: Bplowman-at-pacific.edu
Name: Barbara L. Plowman

Organization: Univ. of the Pacific AADugoni School of Dentistry

Title-Subject: [Filtered] LRWhite

Message: I used LRW and polymerized it with heat also. It came with
a little packet of white powdered accelerator, but I didn't use it.
It did polymerize with heat, I believe a 60 degree C oven. I have
never used LrGold because it wqas more expensive.

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From: protrain-at-emcourses.com
Date: Wed, 26 May 2010 05:01:04 -0500
Subject: [Microscopy] LaB6 filament change procedure for Carl-Zeiss SEM

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Email: dhorne-at-interchange.ubc.ca
Name: Derrick Horne

Organization: UBC BioImaging Facility

Title-Subject: [Filtered] quantitative EBIC Auxiliary Video Amplifier for S4700

Message: The search continues for some absorbed current data.

I believe we've sourced an external signal amplifier that will work,
and now we need an Auxiliary Video Amplifier for our Hitachi S-4700.

With the idea of saving the researcher some money, does anybody have
any idea where I might be able to track one down?

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From riadh.benaissa-at-snclavalin.com Tue May 25 21:40:15 2010
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Dear Listers,

I remember there existing a soft-copy procedure around for this job. I
would appreciate if someone can e-mail me a copy. I just can't find my
copy.

Best

EB


=================================
Erman Bengu

Assistant Professor of Chemistry
Department of Chemistry
Bilkent University

Mailing Address:
Bilkent University,
Department of Chemistry,
06800, Bilkent, Ankara
Turkey

Office: SB #311
E-mail: bengu_AT_fen.bilkent.edu.tr
Phone (Office): +90 (312) 290-2153
(Lab1): +90 (312) 290-2663
(Lab2): +90 (312) 290-3332
Fax: +90 (312) 266-4068
Web: http://www.fen.bilkent.edu.tr/~bengu
==================================


==============================Original Headers==============================
11, 22 -- From bengu-at-fen.bilkent.edu.tr Wed May 26 03:28:08 2010
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11, 22 -- Subject: LaB6 filament change procedure for Carl-Zeiss SEM
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From brgn-at-cimbria.com Wed May 26 04:06:35 2010
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Hi

I hope I am able to help, I guess the main problem is manipulation of the
vacuum system?

Once the Gun Chamber is closed, having had a new filament fitted, allow the
system to pump down with the manifold valve, found at the rear of the gun
chamber, OPEN. IGP remains off.

Whilst this is happening from the SEM Control drop down check under GUN that
LaB6 is selected along with Partial Vent on Stan by under VACUUM.

Pump the complete system until you have a System Vacuum level indicated in
the 007Torr area, then switch IGP on. Watch the IGP or Gun Vacuum and the
System Vacuum to see when the IGP starts to dominate after its initial out
gassing period. As soon as the Gun Vacuum is superior to the System Vacuum
close the manifold valve behind the gun chamber. Wait whilst the Gun Vacuum
stabilises then you are ready to go.

Note the original LaB6 filament setting. During the following procedure
watch the Gun Vacuum and hold an action if the vacuum level deteriorates.
Turn down the filament target and apply the EHT at 1kV progressing to 10kV
at 1kV per minute. On reaching 10kV bring the filament supply up very
slowly, minutes, to a point just below the earlier LaB6 setting. Hold at
this level until the Gun Vacuum is stable and then you may progress steadily
to higher kV if you wish, but always watching the Gun vacuum in the early
hours of LaB6 use.

After this initial period set the appropriate gun saturation point and then
allow the instrument to run up the filament as required.

I hope this helps?

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: bengu-at-fen.bilkent.edu.tr [mailto:bengu-at-fen.bilkent.edu.tr]
Sent: 26 May 2010 09:30
To: protrain-at-emcourses.com


Dear Listers,

I remember there existing a soft-copy procedure around for this job. I
would appreciate if someone can e-mail me a copy. I just can't find my
copy.

Best

EB


=================================
Erman Bengu

Assistant Professor of Chemistry
Department of Chemistry
Bilkent University

Mailing Address:
Bilkent University,
Department of Chemistry,
06800, Bilkent, Ankara
Turkey

Office: SB #311
E-mail: bengu_AT_fen.bilkent.edu.tr
Phone (Office): +90 (312) 290-2153
(Lab1): +90 (312) 290-2663
(Lab2): +90 (312) 290-3332
Fax: +90 (312) 266-4068
Web: http://www.fen.bilkent.edu.tr/~bengu
==================================


==============================Original Headers==============================
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From: holpc-at-firstenergycorp.com
Date: Wed, 26 May 2010 05:57:24 -0500
Subject: [Microscopy] Manual for Amray 1200

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

Hello,

I work with two Philips CM series TEMs.

I am trying to help diagnose some filament life issues, and I need to know
what the temperature I am heating the filament to is, in a real unit
(kelvin), instead of just knowing it is heating stage 21. Does anyone know
of a conversion procedure? The same question for the vacuum readings, the
emission current scale, and is it possible to give meaningful units to what
the emission level setting?

Thanks,


Ben



--
Imaging Technician
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271866
{http://mrcanu.pharm.ox.ac.uk/}



==============================Original Headers==============================
11, 29 -- From ben.micklem-at-pharm.ox.ac.uk Wed May 26 05:23:33 2010
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11, 29 -- From: Ben Micklem {ben.micklem-at-pharm.ox.ac.uk}
11, 29 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
11, 29 -- Date: Wed, 26 May 2010 11:23:28 +0100
11, 29 -- Subject: Philips CM heating and vacuum levels in SI units
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From kristen-at-myautumnproperties.com Wed May 26 05:29:32 2010
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Recently a friend got an Amray 1200 for tinkering on at home, and he would
like to get hold of a manual and/or schematic set. If anyone has one
available, it would be much appreciated. Please contact me on or off list,
and I will pass the information along.

Chris Holp
BETA Labs
Mayfield Village OH 44143


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==============================Original Headers==============================
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From: protrain-at-emcourses.com
Date: Wed, 26 May 2010 06:15:24 -0500
Subject: [Microscopy] Philips CM heating and vacuum levels in SI units

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

Hi

May I help on filament life issues? I am not too familiar with the CM
Series but I would have thought they provide an emission current (beam
current) meter as do other manufacturers. However I do not know any
manufacturer in the EM business who even guess at the filament temperature
under the wide range of operating conditions offered.

What puzzles me with your question is the heat applied to the filament is
just part of the gun saturation process, filament current , bias setting
(emission current required) and filament position being the three factors
which relate to the saturation point.

I have trouble shooted filament problems on EMs for over 40 years and have
never had cause to relate to actual filament temperature so may I offer some
ideas?

1. As the filament ages it becomes thinner and will eventually break at
the side which is slightly thinned during the bend. The normal break in a
TEM should be through a gentle taper being shown on the failure area.

2. A poor vacuum in the gun will cause high levels of evaporation and
contamination and the shortening of the filament life. Do not take notice of
the vacuum gauge reading as this is almost certainly well away from the gun
area.

3. From time to time ALL manufacturers suffer from being supplied with
poor quality wire made up of metal and a considerable amount of rubbish.
Short life, a smelly chamber and a very dirty cathode assembly are
indications of this problem.

4. Finally we have situations where the operator just does not know
what they are doing when saturating the gun and they overheat the filament.

I wrote an article for Microscopy Today some years ago that related to "The
Life and Death of the Tungsten Hairpin Filament" which may add information
to your fault finding procedures?

Good luck

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com





I am n

-----Original Message-----
X-from: ben.micklem-at-pharm.ox.ac.uk [mailto:ben.micklem-at-pharm.ox.ac.uk]
Sent: 26 May 2010 11:24
To: protrain-at-emcourses.com

Hello,

I work with two Philips CM series TEMs.

I am trying to help diagnose some filament life issues, and I need to know
what the temperature I am heating the filament to is, in a real unit
(kelvin), instead of just knowing it is heating stage 21. Does anyone know
of a conversion procedure? The same question for the vacuum readings, the
emission current scale, and is it possible to give meaningful units to what
the emission level setting?

Thanks,


Ben



--
Imaging Technician
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271866
{http://mrcanu.pharm.ox.ac.uk/}



==============================Original Headers==============================
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11, 29 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
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11, 29 -- Subject: Philips CM heating and vacuum levels in SI units
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==============================Original Headers==============================
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From: david.knecht-at-uconn.edu
Date: Thu, 27 May 2010 12:54:21 -0500
Subject: [Microscopy] Picric acid fixation

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

Hi

It's me back again, so keen was I to have you running again I forgot you may
not know the shut down procedure?

Closing down you go to the SEM Control drop down, VACCUM where you need to
switch off the IGP and release the Partial Vent on Standby trip. Go to the
hand valve behind the gun and slowly open it.

You may now VENT the system. The cathode is held in place by one or two
small allan screws.

Good luck

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com




-----Original Message-----
X-from: bengu-at-fen.bilkent.edu.tr [mailto:bengu-at-fen.bilkent.edu.tr]
Sent: 26 May 2010 09:30
To: protrain-at-emcourses.com


Dear Listers,

I remember there existing a soft-copy procedure around for this job. I
would appreciate if someone can e-mail me a copy. I just can't find my
copy.

Best

EB


=================================
Erman Bengu

Assistant Professor of Chemistry
Department of Chemistry
Bilkent University

Mailing Address:
Bilkent University,
Department of Chemistry,
06800, Bilkent, Ankara
Turkey

Office: SB #311
E-mail: bengu_AT_fen.bilkent.edu.tr
Phone (Office): +90 (312) 290-2153
(Lab1): +90 (312) 290-2663
(Lab2): +90 (312) 290-3332
Fax: +90 (312) 266-4068
Web: http://www.fen.bilkent.edu.tr/~bengu
==================================


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==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
(Not to me - Oshel)
==========================================================

Listers,
I've sent some ideas, but there should be many more floating around
in the microworld.
Note: Please reply to Lampert! Not just to the list, or he won't see
your reply. List + Lampert is perhaps best.

} Date: Tue, 25 May 2010 14:21:33 -0700 (PDT)
} From: Michael Lampert {mlampert-at-aol.com}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Tuesday, May 25, 2010
} at 02:21:28 PM.
}
} realname - Michael Lampert
} Email - mlampert-at-aol.com
} ORGANIZATION - West Salem High School
} EDUCATION - 9-12th Grade High School
} LOCATION - Salem OR
} SUBJECT_OF_QUESTION - Physics
} QUESTION - I am looking for research opportunities for my high
} school kids using my Olympus Polarizing Microscope. I was
} wondering if anyone would have any ideas for students to pursue a
} project that could lead to the Intel Science Talent Search. I coach
} a research team.
}
} also, are there opportunities for research in microscopes for
} graduate students? Are there schools that specialize in this as a
} career?
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576
Microscopy Society of America
Ask a Microscopist
http://www.microscopy.org/microscopy/ask.cfm

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From tobie-at-hargray.com Wed May 26 08:45:38 2010
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Thanks for your reply Steve. I'm using LaB6 and CeB6 filaments, not
Tungsten. They prematurely failure at around 700 hours, when the
manufacturer says they should be lasting 2000+ hours.

The manufacturer has been helpful so far, and I was just wanting to answer
their questions about operating environment. They may not be that familiar
with TEM applications.

I have put up images on our web site of some filaments after they were
removed after becoming too dim to be usable.

http://mrcanu.pharm.ox.ac.uk/failures/

We increase the bias setting as the filament dims over its life, usually
requiring an increase in heating current to maintain saturation- I think
this is why most fail eventually from the insulators melting. But I don't
know how else to maintain the required beam strength when the filament
becomes dimmer. The emission current is always fairly low- again I only have
the scale the microscope has, but it is around the 5% mark for most of its
life, then this may increase to a maximum of 10% when it has got really dim,
and we are on a high bias setting (e.g. 4 out of 6- see how useful these
numbers are to a non-Philips user!).

We are using 500 µm wehnelt apertures, and set the filament tip 150 µm below
the outer surface of the wehnelt aperture. I have tried using a greater
distance to the wehnelt, but the beam was not strong enough for our uses.

We recently tried CeB6, but they had about the same life as LaB6.

Anyone using LaB6 or CeB6 with long life on CMs (or other Philips/FEI TEMs)
for biological applications, and care to share their operating conditions
and filament lifetimes?

I don't think we have unusual requirements for beam intensity, so I am
puzzled as to why the filaments are not lasting as long as the manufacture
claims.


Thanks,


Ben

--
Imaging Technician
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271866
{http://mrcanu.pharm.ox.ac.uk/}



On 26/05/2010 12:15, "Steve Chapman" {protrain-at-emcourses.com} wrote:

} Hi
}
} May I help on filament life issues? I am not too familiar with the CM
} Series but I would have thought they provide an emission current (beam
} current) meter as do other manufacturers. However I do not know any
} manufacturer in the EM business who even guess at the filament temperature
} under the wide range of operating conditions offered.
}
} What puzzles me with your question is the heat applied to the filament is
} just part of the gun saturation process, filament current , bias setting
} (emission current required) and filament position being the three factors
} which relate to the saturation point.
}
} I have trouble shooted filament problems on EMs for over 40 years and have
} never had cause to relate to actual filament temperature so may I offer some
} ideas?
}
} 1. As the filament ages it becomes thinner and will eventually break at
} the side which is slightly thinned during the bend. The normal break in a
} TEM should be through a gentle taper being shown on the failure area.
}
} 2. A poor vacuum in the gun will cause high levels of evaporation and
} contamination and the shortening of the filament life. Do not take notice of
} the vacuum gauge reading as this is almost certainly well away from the gun
} area.
}
} 3. From time to time ALL manufacturers suffer from being supplied with
} poor quality wire made up of metal and a considerable amount of rubbish.
} Short life, a smelly chamber and a very dirty cathode assembly are
} indications of this problem.
}
} 4. Finally we have situations where the operator just does not know
} what they are doing when saturating the gun and they overheat the filament.
}
} I wrote an article for Microscopy Today some years ago that related to "The
} Life and Death of the Tungsten Hairpin Filament" which may add information
} to your fault finding procedures?
}
} Good luck
}
} Steve Chapman FRMS
} Senior Consultant Protrain
} For consultancy and training in electron microscopy world wide
} Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
} www.emcourses.com
}
}
}
}
}
} I am n
}
} -----Original Message-----
} From: ben.micklem-at-pharm.ox.ac.uk [mailto:ben.micklem-at-pharm.ox.ac.uk]
} Sent: 26 May 2010 11:24
} To: protrain-at-emcourses.com
} Subject: [Microscopy] Philips CM heating and vacuum levels in SI units
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hello,
}
} I work with two Philips CM series TEMs.
}
} I am trying to help diagnose some filament life issues, and I need to know
} what the temperature I am heating the filament to is, in a real unit
} (kelvin), instead of just knowing it is heating stage 21. Does anyone know
} of a conversion procedure? The same question for the vacuum readings, the
} emission current scale, and is it possible to give meaningful units to what
} the emission level setting?
}
} Thanks,
}
}
} Ben



==============================Original Headers==============================
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20, 30 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
20, 30 -- Date: Wed, 26 May 2010 16:44:55 +0100
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From c.sherm-at-indyrr.com Wed May 26 11:10:31 2010
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I have been using the LaB6 mini-Vogel mount you show for many years on a CM30 300KV TEM. My observations are:
1. Almost all failures are operator induced. Generally failure is due to heating to rapidly. The LaB6 chip is very brittle and subject to damage by thermo shock. Do your routine run up as you would a new filament, i.e. Slowly.
2. The LaB6 is very sensitive to the vacuum level. Evaporation is markedly increased with a slightly degraded vacuum level. The most likely leak is SF6 which is most easily detected by the smell when you open the gun chamber and by the amount and color of the deposits on the anode.
3. Improper conditioning may result in a hard flash over. Consider the voltage and the HV cable capacitance and you can see a large quick discharge carries a high energy density, resulting in actual filament shift or work function damage. Either way it requires mechanical realignment of the tip or most of your beam will be coming from the side facets rather than the tip.

Of coarse there are a lot of other possibilities but these are the main culprits in our facility. To answer your direct question, we expect and get about 2 years per filament.

Sincerely
Fran Laabs
Ames Laboratory, ISU

________________________________________
X-from: ben.micklem-at-pharm.ox.ac.uk [ben.micklem-at-pharm.ox.ac.uk]
Sent: Wednesday, May 26, 2010 10:46 AM
To: fclaabs-at-iastate.edu

Thanks for your reply Steve. I'm using LaB6 and CeB6 filaments, not
Tungsten. They prematurely failure at around 700 hours, when the
manufacturer says they should be lasting 2000+ hours.

The manufacturer has been helpful so far, and I was just wanting to answer
their questions about operating environment. They may not be that familiar
with TEM applications.

I have put up images on our web site of some filaments after they were
removed after becoming too dim to be usable.

http://mrcanu.pharm.ox.ac.uk/failures/

We increase the bias setting as the filament dims over its life, usually
requiring an increase in heating current to maintain saturation- I think
this is why most fail eventually from the insulators melting. But I don't
know how else to maintain the required beam strength when the filament
becomes dimmer. The emission current is always fairly low- again I only have
the scale the microscope has, but it is around the 5% mark for most of its
life, then this may increase to a maximum of 10% when it has got really dim,
and we are on a high bias setting (e.g. 4 out of 6- see how useful these
numbers are to a non-Philips user!).

We are using 500 µm wehnelt apertures, and set the filament tip 150 µm below
the outer surface of the wehnelt aperture. I have tried using a greater
distance to the wehnelt, but the beam was not strong enough for our uses.

We recently tried CeB6, but they had about the same life as LaB6.

Anyone using LaB6 or CeB6 with long life on CMs (or other Philips/FEI TEMs)
for biological applications, and care to share their operating conditions
and filament lifetimes?

I don't think we have unusual requirements for beam intensity, so I am
puzzled as to why the filaments are not lasting as long as the manufacture
claims.


Thanks,


Ben

--
Imaging Technician
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271866
{http://mrcanu.pharm.ox.ac.uk/}



On 26/05/2010 12:15, "Steve Chapman" {protrain-at-emcourses.com} wrote:

} Hi
}
} May I help on filament life issues? I am not too familiar with the CM
} Series but I would have thought they provide an emission current (beam
} current) meter as do other manufacturers. However I do not know any
} manufacturer in the EM business who even guess at the filament temperature
} under the wide range of operating conditions offered.
}
} What puzzles me with your question is the heat applied to the filament is
} just part of the gun saturation process, filament current , bias setting
} (emission current required) and filament position being the three factors
} which relate to the saturation point.
}
} I have trouble shooted filament problems on EMs for over 40 years and have
} never had cause to relate to actual filament temperature so may I offer some
} ideas?
}
} 1. As the filament ages it becomes thinner and will eventually break at
} the side which is slightly thinned during the bend. The normal break in a
} TEM should be through a gentle taper being shown on the failure area.
}
} 2. A poor vacuum in the gun will cause high levels of evaporation and
} contamination and the shortening of the filament life. Do not take notice of
} the vacuum gauge reading as this is almost certainly well away from the gun
} area.
}
} 3. From time to time ALL manufacturers suffer from being supplied with
} poor quality wire made up of metal and a considerable amount of rubbish.
} Short life, a smelly chamber and a very dirty cathode assembly are
} indications of this problem.
}
} 4. Finally we have situations where the operator just does not know
} what they are doing when saturating the gun and they overheat the filament.
}
} I wrote an article for Microscopy Today some years ago that related to "The
} Life and Death of the Tungsten Hairpin Filament" which may add information
} to your fault finding procedures?
}
} Good luck
}
} Steve Chapman FRMS
} Senior Consultant Protrain
} For consultancy and training in electron microscopy world wide
} Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
} www.emcourses.com
}
}
}
}
}
} I am n
}
} -----Original Message-----
} From: ben.micklem-at-pharm.ox.ac.uk [mailto:ben.micklem-at-pharm.ox.ac.uk]
} Sent: 26 May 2010 11:24
} To: protrain-at-emcourses.com
} Subject: [Microscopy] Philips CM heating and vacuum levels in SI units
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hello,
}
} I work with two Philips CM series TEMs.
}
} I am trying to help diagnose some filament life issues, and I need to know
} what the temperature I am heating the filament to is, in a real unit
} (kelvin), instead of just knowing it is heating stage 21. Does anyone know
} of a conversion procedure? The same question for the vacuum readings, the
} emission current scale, and is it possible to give meaningful units to what
} the emission level setting?
}
} Thanks,
}
}
} Ben



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From sdume-at-plazalama.com Wed May 26 11:43:35 2010
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Ben,

Being a LaB6 user on a different brand of SEM, I didn't think that I had
any meaningful input regarding your issue, until I saw your pictures of
the failed filaments!

Those are FEI filaments, right?? CamScan tried FEI filaments in our
instrument (2040SL) when new. They never would last. Those resistive
heating blocks seemed to have long-term issues maintaining proper
electrical contact. Tried several filaments; they all ended up failing
prematurely.

Went back to using filaments from Kimball Physics. Never a problem since.
Main reason for filament changes now is routine service every 1 - 1.5
years.

Rick


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From kschristensen-at-meistermedia.com Wed May 26 14:12:26 2010
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Email: m_raven-at-lifesci.ucsb.edu
Name: Mary Raven

Organization: University of California, Santa Barbara

Title-Subject: [Filtered] LM: Olympus AX70

Message: Dear Listers

I have inherited an Olympus AX70 microscope in good working
condition. It has an infinity-corrected optical system and is
equipped with a fluorescent burner, motorized turret and nosepiece
but it lacks objectives and filter cubes. At present it either needs
a good home (for the right price) or to be put into service in my
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I'm struggling with the decision to re-equip the microscope or to sell it.

Do any of the facility managers out there have suggestions? Any
thoughts on the sale price for the system as it stands? I haven't
seen many AX70s for sale so it is tough to gauge its worth.

Best Regards
Mary

Mary Raven
Microscopy Facility Director
Neuroscience Research Institute&
Molecular, Cellular& Developmental Biology
The University of California
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From kscott-at-kristenscott.com Wed May 26 20:40:43 2010
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We've just bought a Zeiss LEO1455 VP SEM (tungsten filament) at a gov't auction and are trying to get it up and running. We have a number of other Zeiss SEM's (all FE 1530's and Ultras) and are somewhat familiar with the software and family of tools but don't have experience with this one.

We have it pumping, can get filament current and think we have the detector's configured properly, but there is no beam/signal. The column also gets quite warm but it does not have obvious cooling lines like our other SEMs (does it have cooling that might be plugged?).

Does someone have the same tool that might be able to answer some basic questions? We're not even sure if we have all the parts (it was missing a pump and other stuff?) and some quick pictures of the plumbing would be very helpful. Same thing with the 'low KV anode'- if we open up the gun how do we recognize what we have?

Does anyone have a source for technical documents for this tool? The online help leaves much to be desired.

thanks,

Chris Pawlowicz, B.Eng
Manager, Lab Development
UBM TechInsights
3000 Solandt Rd Ottawa ON Canada K2K 2X2
T: +1 613 599 5145 x4454
F: +1 613 599 6501
E: cpawlowicz-at-ubmtechinsights.com


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10, 22 -- Subject: SEM Zeiss/LEO 1455VP need technical help
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From resumes-at-qpscanada.com Thu May 27 09:21:26 2010
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X-from: Thomas J Burdsall [mailto:tburdsall-at-3dleap.com]
Hello Chris,
see my comments below . I dont have much time to
{snip}
Thom

} We have it pumping, can get filament current (( do you get Emission
} curent?))

I get an reading of Beam Current (250uA) but wasn't sure if this was measured or derived mathematically somehow.

} detector's configured properly, but there is no beam/signal. The
} column also gets quite warm (( Possible TOO Much current.))but it does
} not have obvious cooling lines like our other SEMs (does it
} have cooling that might be plugged?). NO. How hot is it.

After a few hours of running the tool the bottom of the gun gets too warm to comfortably hold your hand on it.




-----Original Message-----
X-from: Xiang Yang [mailto:xiang.yang-at-SMU.CA]

} I have a LEO1450VP SEM in the lab and I think it may be help with your SEM.

} Certainly we have a water chiller providing cooling water to the SEM. Look at the back (bottom) of your instrument; if you see brass openings with water in/out labels, you need a chiller as well.

Yes, we definitely have water chilling hooked up to the tool and running, what I was not sure about was if there were cooling lines running to the gun / around the lenses. On our other SEMs they are external and obvious. In the 1455 there are no external lines that we can see. I was wondering if they were internal. (Thom's reply indicates that it does not above)


} On our lab website (http://fgsr.smu.ca/emc/res_how_sem.html), there are some operational instructions available on LEO1450VP system. You can check it out. If you have more questions, feel free to contact me.
} Best Regards
---------------------------------
} Xiang (Sean) Yang

That's great! I've had a quick look and the documents are excellent to make sure we're not doing something completely wrong :)

thanks,

Chris Pawlowicz

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==============================Original Headers==============================
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From kristin-at-grandeventrentals.com Thu May 27 10:51:58 2010
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I am trying to find information about the chemistry of picric acid fixation. Specifically, I want to know what functional groups are cross linked and what the pH dependence of the reaction is. Any ideas on where I might find this? Thanks- Dave

Dr. David Knecht
Department of Molecular and Cell Biology
Co-head Flow Cytometry and Confocal Microscopy Facility
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)




==============================Original Headers==============================
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From: muske-dukes-at-bendres.com
Date: Thu, 27 May 2010 17:07:46 -0500
Subject: [Microscopy] viaWWW: Number of TEMs

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

David,

One good source for this sort of information is "Histological &
Histochemical Methods: Theory & Practice" by J.A. Kiernan.
Kiernan states that picric acid is a coagulating fixative that works
by binding with basic groups of proteins, and precipitating them. pH
1.5 - 2. The precipitation is reversed if the solution pH is
neutralized.

Phil

} I am trying to find information about the chemistry of picric acid
} fixation. Specifically, I want to know what functional groups are
} cross linked and what the pH dependence of the reaction is. Any
} ideas on where I might find this? Thanks- Dave
}
} Dr. David Knecht
} Department of Molecular and Cell Biology
} Co-head Flow Cytometry and Confocal Microscopy Facility
} U-3125
} 91 N. Eagleville Rd.
} University of Connecticut
} Storrs, CT 06269
} 860-486-2200
} 860-486-4331 (fax)

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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Email: muske-dukes-at-bendres.com
Name: Annie Muske-Dukes

Organization: Bend Research

Title-Subject: [Filtered] Number of TEMs

Message: Hi everyone,

We have school groups come through our facility on occasion for a
tour, and recently a student asked a question that I couldn't answer.
I was hoping someone on here might have a guess.

She was wondering how many TEMs are currently operating in the world,
or even just the United States. It seems like many large universities
have at least one TEM, but that would leave out any non-academic
facilities so I'm not sure if that would give a good estimate.

Does anyone have any idea what the number of TEMs might be? I'm
rather intrigued by the question myself, if I called manufacturers
would they be likely to give me any sales figures?

Thanks,
Annie Muske-Dukes
Bend Research, Bend OR US


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From: Rob.Bowen-at-caddock.com
Date: Tue, 1 Jun 2010 09:40:49 -0500
Subject: [Microscopy] Re: pdf publication policies

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Email: adel-at-halcyonmolecular.com
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Organization: Halcyon Molecular

Title-Subject: [Filtered] Microscopist opening

Message: Halcyon Molecular is seeking a talented microscopist to take
ownership of operating and maintaining electron microscopes, and
trouble shoot needed parameters for obtaining high quality image
data. The following experience is useful: TEM, diffraction, STEM, SE
and BSE imaging in the SEM, x-ray spectroscopy, AFM, STM, basic
experience with UHV. Ideal candidate will thrive in a fast-paced
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process-oriented and being able to learn more complex operations
(like aberration-corrected STEM).

Please send your inquiries and CVs to futures-at-halcyonmolecular.com

Halcyon Molecular is a start-up company located in Redwood city CA.
We have developed a nanorobotic DNA manipulation technology that will
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orders of magnitude on speed, cost, and quality. This development
will transform biology into an information science and make
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From sean-at-metroblogging.com Thu May 27 17:38:30 2010
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Dear All

A client wishes to embark on a study which requires marking a series of lines through features of a digital image and then measuring the angles of those lines to determine the degree of deviation from parallel.

Could you suggest software that could achieve this?

Many thanks in advance,


Naomi McCallum
Supervising Scientist, EM Unit, Anatomical Pathology
Pathology Queensland
_________________________________________________
Clinical and Statewide Services Division| Queensland Health

Block 7 Level 2
RBWH Queensland 4029
Ph: 07 3636 8057
Mob:
Fax: 07 3636 8908
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11, 25 -- From prvs=17673255a6=naomi_mccallum-at-health.qld.gov.au Mon May 31 01:14:29 2010
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From camd-at-cutterbuck.com Mon May 31 02:18:39 2010
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Hello Naomi,

What you are asking for is pretty straight forward. I would suggest to look
at iTEM or Scandium (depending on the instrument). You can find the links
here: http://www.resaltatech.com/software_overview.htm. The base versions of
both software packages have manual measurements included. You can measure
angles of lines and transfer them to a spreadsheet (Excel) for further
analysis, or you can get statistical information (for example mean value and
standard deviation of the angles) directly in the software.

Let me know if you have any questions.

Mike Bode

---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode at ResAltaTech.com
www.ResAltaTech.com





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{zaluzec-at-microscopy.com} ; Mon, 31 May 2010 12:24:03 -0300

Hi to all,

I am sorry if this message does not talk about microscopy but I have a question about the publication and duplication policies of scientific papers.
When I was student (in my head it was yesterday but it may be a little bit earlier), before the pdf era, when we wanted a paper we had preprinted cards that we sent to the authors and there was a kind of unwritten law that wanted that the authors graciously send a copy of their paper (however if they did not, you couldn't sue them ;-)).
Now the publications are online and if we want a paper we have to graciously pay something between 30 and 50 dollars.
But what about the good old unwritten law? Is it still possible to send an email to an author and ask for a pdf copy of his paper or do we really have to translate everything down on earth in $$?

Regards,

Stephane




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From resa-at-festival-autrans.com Tue Jun 1 03:15:07 2010
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Hi, Stephane,
I've been grumbling about the $30 for a pdf, too. However, I realized
that the big money is if you want the article RIGHT NOW!!!! If I want to
get up from my chair, trundle down to the local library and fill out a paper
form with the citation info, and pay an interlibrary loan fee of $2 or $3,
and wait several days, I can get an adequate quality copy from someplace
that has the publication. The. Inertia still keeps me from getting a lot of
articles this way, but I figure having some tax money go to the library is
definitely a bet buy.

Rob Bowen

} From: {nizets2-at-yahoo.com}
} Reply-To: {nizets2-at-yahoo.com}
} Date: Tue, 1 Jun 2010 02:19:24 -0500
} To: {rob.bowen-at-caddock.com}
} Subject: [Microscopy] pdf publication policies
}
}
}
}
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
}
} Hi to all,
}
} I am sorry if this message does not talk about microscopy but I have a
} question about the publication and duplication policies of scientific papers.
} When I was student (in my head it was yesterday but it may be a little bit
} earlier), before the pdf era, when we wanted a paper we had preprinted cards
} that we sent to the authors and there was a kind of unwritten law that wanted
} that the authors graciously send a copy of their paper (however if they did
} not, you couldn't sue them ;-)).
} Now the publications are online and if we want a paper we have to graciously
} pay something between 30 and 50 dollars.
} But what about the good old unwritten law? Is it still possible to send an
} email to an author and ask for a pdf copy of his paper or do we really have to
} translate everything down on earth in $$?
}
} Regards,
}
} Stephane
}
}
}
}
} ==============================Original Headers==============================
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==============================Original Headers==============================
5, 21 -- From Rob.Bowen-at-caddock.com Tue Jun 1 09:40:49 2010
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From: kenconverse-at-qualityimages.biz
Date: Tue, 1 Jun 2010 11:14:01 -0500
Subject: [Microscopy] Re: Measurement software

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

On Mon, May 31, 2010 at 2:16 AM, {naomi_mccallum-at-health.qld.gov.au} wrote:

} Dear All
}
} A client wishes to embark on a study which requires marking a series of lines through features of a digital image and then measuring the angles of those lines to determine the degree of deviation from parallel.
}
} Could you suggest software that could achieve this?
}
} Many thanks in advance,
}
}
} Naomi McCallum
} Supervising Scientist, EM Unit, Anatomical Pathology
} Pathology Queensland
} _________________________________________________
} Clinical and Statewide Services Division| Queensland Health
}
} Block 7 Level 2
} RBWH  Queensland 4029
} Ph: 07 3636 8057
} Mob:
} Fax: 07 3636 8908
} Email: naomi_mccallum-at-health.qld.gov.au
} Web: http://www.health.qld.gov.au/qhcss/qhps
}


Naomi,

I always suggest ImageJ, which is very capable, can be customized/extended
by the user with macros and plugins (depending on users programming skills
and desired effort) and is distributed freely by the US National Institutes
of Health. Additionally, there is a fantastic user community and listserver
to assist the novice.

What you described should be trivial for just about any image analysis
program. It certainly is for ImageJ. A macro could be recorded to help,
but it can be done relatively easily doing each step manually.

http://rsbweb.nih.gov/ij/

Commercial software certainly has a place, particularly if an end user
requires support or hardware interfacing. Those are areas where it can be
worth paying for the knowledge. For simple tasks, though, especially if
it's a one-time project, I like the economical choice. No sense buying a
bulldozer when a shovel will do the job.


Jim


----------------------------------------------
Jim Passmore
Research Associate
Sealed Air Corporation
james.passmore-at-sealedair.com
864-433-2927 voice
864-433-2205 fax
----------------------------------------------


==============================Original Headers==============================
13, 22 -- From james.passmore-at-sealedair.com Tue Jun 1 10:33:23 2010
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13, 22 -- From: James Passmore {james.passmore-at-sealedair.com}
13, 22 -- Date: Tue, 1 Jun 2010 11:33:01 -0400
13, 22 -- Message-ID: {AANLkTinWzLCcd_BLkAbtriRto89AppmXo_MfvfNZvD8g-at-mail.gmail.com}
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From sdikeman-at-thirousa.com Tue Jun 1 10:53:56 2010
Return-Path: {sdikeman-at-thirousa.com}
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But Jim, the bulldozer is so much more fun!

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: james.passmore-at-sealedair.com [mailto:james.passmore-at-sealedair.com]
Sent: Tuesday, June 01, 2010 11:35 AM
To: kenconverse-at-qualityimages.biz

On Mon, May 31, 2010 at 2:16 AM, {naomi_mccallum-at-health.qld.gov.au} wrote:

} Dear All
}
} A client wishes to embark on a study which requires marking a series of
lines through features of a digital image and then measuring the angles of
those lines to determine the degree of deviation from parallel.
}
} Could you suggest software that could achieve this?
}
} Many thanks in advance,
}
}
} Naomi McCallum
} Supervising Scientist, EM Unit, Anatomical Pathology
} Pathology Queensland
} _________________________________________________
} Clinical and Statewide Services Division| Queensland Health
}
} Block 7 Level 2
} RBWH  Queensland 4029
} Ph: 07 3636 8057
} Mob:
} Fax: 07 3636 8908
} Email: naomi_mccallum-at-health.qld.gov.au
} Web: http://www.health.qld.gov.au/qhcss/qhps
}


Naomi,

I always suggest ImageJ, which is very capable, can be customized/extended
by the user with macros and plugins (depending on users programming skills
and desired effort) and is distributed freely by the US National Institutes
of Health. Additionally, there is a fantastic user community and listserver
to assist the novice.

What you described should be trivial for just about any image analysis
program. It certainly is for ImageJ. A macro could be recorded to help,
but it can be done relatively easily doing each step manually.

http://rsbweb.nih.gov/ij/

Commercial software certainly has a place, particularly if an end user
requires support or hardware interfacing. Those are areas where it can be
worth paying for the knowledge. For simple tasks, though, especially if
it's a one-time project, I like the economical choice. No sense buying a
bulldozer when a shovel will do the job.


Jim


----------------------------------------------
Jim Passmore
Research Associate
Sealed Air Corporation
james.passmore-at-sealedair.com
864-433-2927 voice
864-433-2205 fax
----------------------------------------------


==============================Original Headers==============================
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==============================Original Headers==============================
25, 29 -- From kenconverse-at-qualityimages.biz Tue Jun 1 11:14:01 2010
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From: PhillipsT-at-missouri.edu
Date: Tue, 1 Jun 2010 12:36:49 -0500
Subject: [Microscopy] FS inverse contrast

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

Hello all,
I have been performing HPF/FS on cell culture with vaccinia. Tried a number of FS coctails in an attempt to determine the best ultrastructure freeze media. When I use 1% and 2% OsO4 + 0.1%UA in Acetone, the mature virions are electron dense and I couldn’t resolve fine detail.

I recently tried 0.5% OsO4 + 0.1% UA in acetone with the anticipation that the mature virions would be less dense. Surprisingly, the virions had reversed contrast. The FS schedule was -90C 3 days and gradually warmed to 21C, acetone washed 4X and resin infiltrated.

One suggestion was to use 1% OsO4 + 0.1%UA in acetone and acetone wash at -80C instead of leaving in OsO4 for the duration. Can anyone explain why concentration change of the OsO4 resulted in negative contrast?
Other cocktails used were 0.5% GA +0.1%TA in acetone, 2% UA/MeOH in acetone. Both resulted in patches of cytoplasm loss.

Thank you
Karen

==============================Original Headers==============================
4, 31 -- From vau-at-ufl.edu Tue Jun 1 12:09:25 2010
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From brewsternn-at-softeng.com Tue Jun 1 12:33:30 2010
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In an earlier life I did a ton of freeze substitution although never of viruses. My experience is that the osmication reaction is sensitive to subtle triggers at -80 C. Sometimes after FS in acetone, rinsing the now light brown tissues in 100% ethanol several times and then storing in ethanol overnight led to an intense osmication reaction with the tissue and ethanol solution going pitch black.

I don't have a solution for you but you could try FS in tetrahydrofuran. We got startling different views with this solvent compared to acetone or ethanol. It might be worth a try.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: vau-at-ufl.edu [mailto:vau-at-ufl.edu]
Sent: Tuesday, June 01, 2010 12:10 PM
To: Phillips, Thomas E.

Hello all,
I have been performing HPF/FS on cell culture with vaccinia. Tried a number of FS coctails in an attempt to determine the best ultrastructure freeze media. When I use 1% and 2% OsO4 + 0.1%UA in Acetone, the mature virions are electron dense and I couldn't resolve fine detail.

I recently tried 0.5% OsO4 + 0.1% UA in acetone with the anticipation that the mature virions would be less dense. Surprisingly, the virions had reversed contrast. The FS schedule was -90C 3 days and gradually warmed to 21C, acetone washed 4X and resin infiltrated.

One suggestion was to use 1% OsO4 + 0.1%UA in acetone and acetone wash at -80C instead of leaving in OsO4 for the duration. Can anyone explain why concentration change of the OsO4 resulted in negative contrast?
Other cocktails used were 0.5% GA +0.1%TA in acetone, 2% UA/MeOH in acetone. Both resulted in patches of cytoplasm loss.

Thank you
Karen

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From: perdi001-at-umn.edu
Date: Tue, 1 Jun 2010 21:45:05 -0500
Subject: [Microscopy] viaWWW: Reichert-Jung Ultracut E ultra-microtome

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

I asked a colleague who has a similar microscope in a core lab and he
replied:

I think it's relatively worthless for her to sell it.

If she wants to put it into service, it's going to take a fair amount of
cash to add the objectives--let's say $15K minimum--and filter
cubes--$4K. If really gets down to whether she needs another workhorse
microscope in her facility. If yes, then she can put it together for
$20K rather than spending $60K. If she's really needing something else
in the facility, then she has to think about whether she wants to save
that $20K to put towards some other system.



m_raven-at-lifesci.ucsb.edu wrote:
}
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} Email: m_raven-at-lifesci.ucsb.edu
} Name: Mary Raven
}
} Organization: University of California, Santa Barbara
}
} Title-Subject: [Filtered] LM: Olympus AX70
}
} Message: Dear Listers
}
} I have inherited an Olympus AX70 microscope in good working
} condition. It has an infinity-corrected optical system and is
} equipped with a fluorescent burner, motorized turret and nosepiece
} but it lacks objectives and filter cubes. At present it either needs
} a good home (for the right price) or to be put into service in my
} multiuser facility.
}
} I'm struggling with the decision to re-equip the microscope or to sell it.
}
} Do any of the facility managers out there have suggestions? Any
} thoughts on the sale price for the system as it stands? I haven't
} seen many AX70s for sale so it is tough to gauge its worth.
}
} Best Regards
} Mary
}
} Mary Raven
} Microscopy Facility Director
} Neuroscience Research Institute&
} Molecular, Cellular& Developmental Biology
} The University of California
} Santa Barbara, CA 93106-5060
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} 11, 11 -- From zaluzec-at-microscopy.com Wed May 26 19:57:51 2010
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--
Larry Ackerman, Specialist
Electron Microscopy Lab
Manager, DERC Microscopy Core
UCSF, Dept. of Anatomy, Rm S1355
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

415-999-4758

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From don-at-w00f.com Tue Jun 1 14:25:00 2010
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Hello-

A student in my group is using CBED on our Hitachi H-7000 FA TEM. One thing though: we usually take the condenser aperture out to get a large convergence angle for measuring HOLZ lines. But now we are noticing that we can't form a probe smaller than about 500 nm with the CA out, because of a coma-type of distortion that affects the probe shape, so it doesn't form a circular spot. In fact, the problem seems even worse in Fine-Probe mode (which turns the middle condenser lens on). The probe looks fine with the CA in, but the convergence semi-angle is only about 3 mrad, whereas it is 8 mrad with the CA out. Does anyone you know if there is an alignment procedure for an H-7000 that can reduce this problem (e.g., coma-free alignment), or is it just an inherent artifact? I'd like to buy a CA strip with some bigger holes than what we have, but it's hard to know in advance if that would fix this. Thanks for any input.

-Phil
------------------------------------------
Phil Ahrenkiel, Assistant Professor
Nanoscience and Nanoengineering Ph.D. Program
South Dakota School of Mines and Technology
501 E. Saint Joseph St.
Rapid City, SD 57701 U.S.A.
Office: EP 221
Phone: 605-394-5238, Fax: 605-394-2365
Email: Phil.Ahrenkiel-at-sdsmt.edu




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From mandy.olaker-at-libertysavingsbank.com Tue Jun 1 17:33:38 2010
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Dear All

Many thanks for your helpful responses. The consensus appears to be ImageJ with a couple of other suggestions also.
My next challenge is with the red tape of a government run IT departments to allow this software to be used.

Thanks again
Naomi

} } } {naomi_mccallum-at-health.qld.gov.au} 31/05/2010 4:25 pm } } }



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Dear All

A client wishes to embark on a study which requires marking a series of lines through features of a digital image and then measuring the angles of those lines to determine the degree of deviation from parallel.

Could you suggest software that could achieve this?

Many thanks in advance,


Naomi McCallum
Supervising Scientist, EM Unit, Anatomical Pathology
Pathology Queensland
_________________________________________________
Clinical and Statewide Services Division| Queensland Health

Block 7 Level 2
RBWH Queensland 4029
Ph: 07 3636 8057
Mob:
Fax: 07 3636 8908
Email: naomi_mccallum-at-health.qld.gov.au
Web: http://www.health.qld.gov.au/qhcss/qhps



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Email: perdi001-at-umn.edu
Name: Jorge Perdigao

Organization: University of Minnesota

Title-Subject: [Filtered] Reichert-Jung Ultracut E ultra-microtome

Message: I need a copy (digital or paper) of the manufacturer's
instruction booklet for a Reichert-Jung Ultracut E.

Thank you
JP

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7, 11 -- From zaluzec-at-microscopy.com Tue Jun 1 21:45:05 2010
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From: jfmjfm-at-umich.edu
Date: Thu, 3 Jun 2010 13:23:42 -0500
Subject: [Microscopy] M&M2010: Late Breaking Poster Session now accepting abstracts

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Email: vau-at-ufl.edu
Name: Karen Kelley

Organization: University of Florida

Title-Subject: [Filtered] HPF cocktail result in inverse contrast

Message: Hello all,
I have been performing HPF/FS on cell culture
with vaccinia. Tried a number of FS coctails in
an attempt to determine the best ultrastructure
freeze media. When I use 1% and 2% OsO4 + 0.1%UA
in Acetone, the mature virions are electron dense
and I couldnít resolve fine detail. I recently
tried 0.5% OsO4 + 0.1% UA in acetone with the
anticipation that the mature virions would be
less dense. Surprisingly, the virions had
reversed contrast. The FS schedule was -90C 3
days and gradually warmed to 21C, acetone washed
4X and resin infiltrated. One suggestion was to
use 1% OsO4 + 0.1%UA in acetone and acetone wash
at -80C instead of leaving in OsO4 for the
duration. Can anyone explain what causes
negative contrast?
Other cocktails used were 0.5% GA +0.1%TA in
acetone, 2% UA/MeOH in acetone. Both resulted in
patches of cytoplasm loss.

Thank you
Karen

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8, 13 -- From zaluzec-at-microscopy.com Tue Jun 1 21:45:52 2010
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From hoang_nguyen-at-mail2world.com Tue Jun 1 22:35:38 2010
Return-Path: {hoang_nguyen-at-mail2world.com}
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Hello Jorge,

Since I can't send attachments to the list I am emailing you off list. I
will reply to the list also as is protocol, so you will see a reply w/o
the attachment.

I am attaching the manual as a "reader" application for Windows (an EXE
file). I am renaming it to a .e-- extension so that the mailer/virus
scanners don't go crazy; please change the extension back to EXE and you
might want to scan it to assure yourself it is not an evil virus.

I also have the original scanned pages I could send as a zip document if
this version does not work for you.

Dale Callaham
Umass -at- Amherst MA

perdi001-at-umn.edu wrote:
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} Email: perdi001-at-umn.edu
} Name: Jorge Perdigao
}
} Organization: University of Minnesota
}
} Title-Subject: [Filtered] Reichert-Jung Ultracut E ultra-microtome
}
} Message: I need a copy (digital or paper) of the manufacturer's
} instruction booklet for a Reichert-Jung Ultracut E.
}
} Thank you
} JP
}
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} ==============================Original Headers==============================
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==============================Original Headers==============================
7, 22 -- From dac-at-research.umass.edu Wed Jun 2 09:03:06 2010
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From relojobs-at-relocationcentral.com Wed Jun 2 09:22:49 2010
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Email: ropope-at-gmail.com
Name: Robert

Title-Subject: [Filtered] Question about Sterility of organisms in Epoxy

Message: Has anyone had dealings with the USDA, CDC, or HHS, or other
agencies with respect to proving sterility of agents that have been
fixed (4%paraformaldehyde/1%gluteraldehyde), osmicated(1% osmium),
(potentially irradiated 2X16[6] rads), dehydrated, and embedded in
epoxy.

I find myself getting questions about things that never were
questioned in the past.

Any and all replies are greatly appreciated.
Robert

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7, 11 -- From zaluzec-at-microscopy.com Wed Jun 2 19:35:15 2010
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From toitimtoi-at-tialia.com Wed Jun 2 20:14:38 2010
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Hi all,

Please note that the Late Breaking Poster Session at the M&M2010 Meeting to be held in Portland, Oregon 1-5th of August 2010, is now open for abstract submission.
Further details and the current list of Late Breaking Posters can be see at:

http://www.microscopy.org/MandM/2010/posters.cfm#Late-Breaking

Text from the website:

The Microscopy and Microanalysis 2010 Executive Program Committee invites submission of abstracts for a special late-breaking poster session at M&M 2010. Submissions are encouraged that include progress in microscopy and microanalysis in physical, biological and analytical science. To have your presentation accepted for the late breaking poster session, you MUST compile a full two page abstract in the accepted M&M Conference Paper Template format and submit it to the Program Chair by close of business (5pm EDT) on the 16th of July 2010. Microsoft Word documents will be accepted, however, PDFs are preferred. The object of the two page abstract is to allow the Executive Program Committee to review the proposed presentation and determine if it meets the standards set by that committee.

--
John Mansfield PhD Cphys CSci MInstP
2010 Microscopy & Microanalysis Program Chair
North Campus Electron Microbeam Analysis Laboratory
417 SRB, University of Michigan
2455 Hayward, Ann Arbor MI 48109-2143 USA
Phone: (734) 936-3352 FAX (734) 763-2282
Cell. Phone: (734) 834-3913
(Leaving a phone message at 936-3352 is preferable to 834-3913)
Email: jfmjfm-at-umich.edu
URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
Location: N 42 ° 17' 38.3" (42 ° 17.6391' ) W 83 ° 42' 38.6" (-83 ° 42.6439')
AIM: thejfmjfm
Yahoo: thejfmjfm
Skype: thejfmjfm

Home address:
4304 Spring Lake Boulevard
Ann Arbor MI 48108-9657
Phone (734) 994-3096
Location: N 42 ° 13' 28.8" (42 ° 13.4807') W 83 ° 45' 47.9" 42° 16' 48" (-83 ° 45.7980')

Please note: Electronic Mail is not secure, but should be read several times every day, and should definitely be used for urgent or sensitive issues.






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13, 18 -- Content-Type: text/plain; charset=iso-8859-1
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13, 18 -- Subject: M&M2010: Late Breaking Poster Session now accepting abstracts
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From: mcauliff-at-umdnj.edu
Date: Thu, 3 Jun 2010 14:44:06 -0500
Subject: [Microscopy] Re: osmium + ferrocyanide and EmBed 812, final answer

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

Dear all:

So I took some more of the same brain tissue and used fresh osmium w/o
ferrocyanide AND I embedded 100 micron Vibratome sections in fresh Spurr
(according to Anne Ellis).
Same result!
Badly fixed brains (by another lab) seems to be the problem, not a rxn
between osmium+ferrocyanide and EmBed 812.

Geoff

mcauliff-at-umdnj.edu wrote:
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} Greetings all:
}
} I embedded routinely processed mouse brain in EmBed 812, no problems.
} The following week I embedded similar tissue processed the same EXCEPT
} that I used an osmium ferrocynide mix for osmication instead of plain
} buffered osmium.
} All other steps were identical.
} The Os-ferrocyanide brain is impossible to cut, acts like it was not
} infiltrated. Pure plastic cuts fine. Is it possible that Os-ferrocyanide
} reacts with something in the EmBed 812??
}
} Geoff
}
}


--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff-at-umdnj.edu
**********************************************



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8, 28 -- References: {201005131726.o4DHQcvB021986-at-ns.microscopy.com}
8, 28 -- In-reply-to: {201005131726.o4DHQcvB021986-at-ns.microscopy.com}
==============================End of - Headers==============================




From: martin.roe-at-nottingham.ac.uk
Date: Fri, 4 Jun 2010 08:28:22 -0500
Subject: [Microscopy] viaWWW: Vac System PB for a Mk II 2000FX

Contents Retrieved from Microscopy Listserver Archives
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Email: martin.roe-at-nottingham.ac.uk
Name: Martin Roe

Organization: Notts. University

Title-Subject: [Filtered] Vac System PB for a Mk II 2000FX.

Message: Hello,
Does anybody have a spare vacuum control board for a JEOL 2000FX Mark
II TEM you are willing to donate or sell? The serial no of the board
is MP002784-00.
We have been trying to source a board for some considerable time now.
Any thoughts or help as to where we may be able to get one would be
greatly appreciated too.
Regards
Martin Roe



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==============================Original Headers==============================
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8, 11 -- Date: Fri, 4 Jun 2010 08:28:19 -0500
8, 11 -- To: microscopy-at-microscopy.com
8, 11 -- From: martin.roe-at-nottingham.ac.uk (by way of MicroscopyListserver)
8, 11 -- Subject: viaWWW: Vac System PB for a Mk II 2000FX
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==============================End of - Headers==============================




From: maimai_huang-at-seed.net.tw
Date: Fri, 4 Jun 2010 08:29:19 -0500
Subject: [Microscopy] viaWWW: About script modification in Multiple EELS Acquisition

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Email: maimai_huang-at-seed.net.tw
Name: Michael R. S. Huang

Organization: material science and engineering department

Title-Subject: [Filtered] About script
modification in Multiple EELS Acquisition

Message: Dear All:

We are searching for a script for multiple EELS
acquisition. As previously mentioned, the script
named ìMultiple EELS Acquisitionî proposed by
David Mitchell seemed to meet our requirements.
However, the script did not work for our
JEOL2100F and Enfina, probably due to the script
originally designed for older machine. Another
script ìKsEELSî, proposed by Dr. Koji Kimoto also
had similar function
(http://www.hremresearch.com/Eng/plugin/plugin001/index.html).Unfortunately,
both scripts have the same problem in our system.
We think as long as they can be modified
slightly, they will do us lots of favors in EELS
acquisitions, especially for eliminating the
effects from environment instability.

We are so bothered by this trouble for a long
period of time and really appreciate that any
script experts can provide us with informative
assistance.

Thanks and Best Regards

Michael R. S. Huang
PhD Student
Material Science and Engineering Department,
National Cheng Kung University, Taiwan
E-mail: maimai_huang-at-seed.net.tw


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==============================Original Headers==============================
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12, 13 -- Subject: viaWWW: About script modification in Multiple EELS Acquisition
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From: maloneyb-at-fiu.edu
Date: Fri, 4 Jun 2010 13:25:54 -0500
Subject: [Microscopy] Olympus BH2 Fluorescent scope

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

Dear Group: does anyone have a copy of the manual for this scope?
Thanks so much
Barbara

==============================Original Headers==============================
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1, 18 -- Date: Fri, 4 Jun 2010 14:25:53 -0400
1, 18 -- From: Barbara Maloney {maloneyb-at-fiu.edu}
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From: rosemary.white-at-csiro.au
Date: Mon, 7 Jun 2010 02:18:23 -0500
Subject: [Microscopy] pdf publication policies

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

I've had requests for pdfs of papers, and am happy to send them, and have
found others happy to email copies of their papers. For example, our
library cannot afford to pay for immediate online access to all journals,
and if I want a pdf of a very recent article, they have to photocopy it and
send that, they cannot send the high quality pdf until the waiting time -
6-12 months - is over. So if it's an article in which good quality image
reproduction is essential, I usually email the author. I could wait for 6
months....but by then I'd have forgotten why I wanted to read it.
cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601

T 02 6246 5475
F 02 6246 5334
E rosemary.white-at-csiro.au

On 2/06/10 12:47 AM, "Rob.Bowen-at-caddock.com" {Rob.Bowen-at-caddock.com} wrote:

}
}
}
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}
} Hi, Stephane,
} I've been grumbling about the $30 for a pdf, too. However, I realized
} that the big money is if you want the article RIGHT NOW!!!! If I want to
} get up from my chair, trundle down to the local library and fill out a paper
} form with the citation info, and pay an interlibrary loan fee of $2 or $3,
} and wait several days, I can get an adequate quality copy from someplace
} that has the publication. The. Inertia still keeps me from getting a lot of
} articles this way, but I figure having some tax money go to the library is
} definitely a bet buy.
}
} Rob Bowen
}
} } From: {nizets2-at-yahoo.com}
} } Reply-To: {nizets2-at-yahoo.com}
} } Date: Tue, 1 Jun 2010 02:19:24 -0500
} } To: {rob.bowen-at-caddock.com}
} } Subject: [Microscopy] pdf publication policies
} }
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} } ----------------------------------------------------------------------------
} }
} } Hi to all,
} }
} } I am sorry if this message does not talk about microscopy but I have a
} } question about the publication and duplication policies of scientific papers.
} } When I was student (in my head it was yesterday but it may be a little bit
} } earlier), before the pdf era, when we wanted a paper we had preprinted cards
} } that we sent to the authors and there was a kind of unwritten law that wanted
} } that the authors graciously send a copy of their paper (however if they did
} } not, you couldn't sue them ;-)).
} } Now the publications are online and if we want a paper we have to graciously
} } pay something between 30 and 50 dollars.
} } But what about the good old unwritten law? Is it still possible to send an
} } email to an author and ask for a pdf copy of his paper or do we really have
} } to
} } translate everything down on earth in $$?
} }
} } Regards,
} }
} } Stephane
} }
} }
} }
} }
} } ==============================Original Headers==============================
} } 6, 25 -- From nizets2-at-yahoo.com Tue Jun 1 02:10:09 2010
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} } 6, 25 -- Date: Tue, 1 Jun 2010 00:10:07 -0700 (PDT)
} } 6, 25 -- From: Stephane Nizet {nizets2-at-yahoo.com}
} } 6, 25 -- Subject: pdf publication policies
} } 6, 25 -- To: microscopy-at-microscopy.com
} } 6, 25 -- MIME-Version: 1.0
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}
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} ==============================Original Headers==============================
} 5, 21 -- From Rob.Bowen-at-caddock.com Tue Jun 1 09:40:49 2010
} 5, 21 -- Received: from msg.caddock.com (206-192-252-70.lsnetworks.net
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} 5, 21 -- User-Agent: Microsoft-Entourage/11.3.3.061214
} 5, 21 -- Date: Tue, 01 Jun 2010 07:42:25 -0700
} 5, 21 -- Subject: Re: [Microscopy] pdf publication policies
} 5, 21 -- From: Rob Bowen {Rob.Bowen-at-caddock.com}
} 5, 21 -- To: {nizets2-at-yahoo.com}
} 5, 21 -- CC: {microscopy-at-microscopy.com}
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==============================Original Headers==============================
7, 41 -- From prvs=7675a41d5=Rosemary.White-at-csiro.au Mon Jun 7 02:18:22 2010
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7, 41 -- Subject: Re: [Microscopy] Re: pdf publication policies
7, 41 -- From: Rosemary White {rosemary.white-at-csiro.au}
7, 41 -- To: "Rob.Bowen-at-caddock.com" {Rob.Bowen-at-caddock.com} ,
7, 41 -- {microscopy-at-microscopy.com}
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From: mshaw1-at-lumc.edu
Date: Mon, 7 Jun 2010 08:28:08 -0500
Subject: [Microscopy] viaWWW: deconvolution & confocal microscopy

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Email: mshaw1-at-lumc.edu
Name: Molly Shaw

Organization: Loyola University Chicago

Title-Subject: [Filtered] deconvolution & confocal microscopy

Message: Hello,

I am a first-year graduate student trying to get started with
colocalization analysis of IF images taken by confocal microscopy. I
am working with fixed cells and mainly trying to measure
colocalization of 2 proteins. My main questions are concerning the
use of z-stacks and deconvolution:
1. Are z-stacks always necessary for measurement of colocalization,
or are single images sufficient?
2. I understand that deconvolution is essential for wide-field
images, but does it improve results that much when you're already
using confocal?
3. If you would recommend deconvolution, is the ImageJ plug-in a good
one to use?

Thank you for your time & help. I have read many articles on this
topic, but I keep ending up with conflicting answers.

Thanks again,
Molly Shaw

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==============================Original Headers==============================
9, 11 -- From zaluzec-at-microscopy.com Mon Jun 7 08:28:08 2010
9, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22])
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9, 11 -- Message-Id: {p06240802c832a4564047-at-[206.69.208.22]}
9, 11 -- Date: Mon, 7 Jun 2010 08:28:06 -0500
9, 11 -- To: microscopy-at-microscopy.com
9, 11 -- From: mshaw1-at-lumc.edu (by way of MicroscopyListserver)
9, 11 -- Subject: viaWWW: deconvolution & confocal microscopy
9, 11 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed"
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From: rhsia-at-umaryland.edu
Date: Mon, 7 Jun 2010 08:28:31 -0500
Subject: [Microscopy] viaWWW: flat embedding mold

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Email: rhsia-at-umaryland.edu
Name: Ru-ching Hsia

Organization: U Maryland EM Core Facility

Title-Subject: [Filtered] flat embedding mold

Message: Hi, everyone,

I would like to get some consumer opinion about flexible flat
embedding molds for spurs resin. We routinely use this type of molds
in our core facility for spurs embedding because they require less
resin and also produce smaller blocks to save storage space. We
recently ordered a batch of flexible embedding molds, none of the
blocks came out sectionable. After investigation, we realized the
new molds are the culprits. We contacted the vendor and were told
that we need to bake the mold in the oven before we use them. They
also said it works better for epon instead of spurs. Of course,
there was no information stating this fact in the catalogue or in the
package (they also refuse to let us return them). After thorough
cleaning and baking, the mold worked a couple of times then the
problem occurred again. Right now, we dare not to risk our clients'
samples using these molds again. If you have a favorite flexible
flat mold that is durable and reliable for spurs resin, I will be
grateful if you would share this information with me.

Thank you so much.

All the best,

Ru-ching

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==============================Original Headers==============================
10, 11 -- From zaluzec-at-microscopy.com Mon Jun 7 08:28:31 2010
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10, 11 -- Date: Mon, 7 Jun 2010 08:28:28 -0500
10, 11 -- To: microscopy-at-microscopy.com
10, 11 -- From: rhsia-at-umaryland.edu (by way of MicroscopyListserver)
10, 11 -- Subject: viaWWW: flat embedding mold
10, 11 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed"
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From: mcauliff-at-umdnj.edu
Date: Mon, 7 Jun 2010 09:55:30 -0500
Subject: [Microscopy] Re: viaWWW: flat embedding mold

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

The blue flat embedding molds do not hold up well to the components in
Spurr resin. I bought mine from Ted Pella and their catalogue states
that they will not last very
long using Spurr. I had to use Spurr (the old, low viscosity formula)
because I was embedding whole zebrafish embryos. After about 3 'cycles'
the molds started to fray at
the edges. I just bought more molds.
The white flexible molds do not seem to have this problem but I could
not find them in the size and configuration I needed.

Geoff


rhsia-at-umaryland.edu wrote:
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}
} Email: rhsia-at-umaryland.edu
} Name: Ru-ching Hsia
}
} Organization: U Maryland EM Core Facility
}
} Title-Subject: [Filtered] flat embedding mold
}
} Message: Hi, everyone,
}
} I would like to get some consumer opinion about flexible flat
} embedding molds for spurs resin. We routinely use this type of molds
} in our core facility for spurs embedding because they require less
} resin and also produce smaller blocks to save storage space. We
} recently ordered a batch of flexible embedding molds, none of the
} blocks came out sectionable. After investigation, we realized the
} new molds are the culprits. We contacted the vendor and were told
} that we need to bake the mold in the oven before we use them. They
} also said it works better for epon instead of spurs. Of course,
} there was no information stating this fact in the catalogue or in the
} package (they also refuse to let us return them). After thorough
} cleaning and baking, the mold worked a couple of times then the
} problem occurred again. Right now, we dare not to risk our clients'
} samples using these molds again. If you have a favorite flexible
} flat mold that is durable and reliable for spurs resin, I will be
} grateful if you would share this information with me.
}
} Thank you so much.
}
} All the best,
}
} Ru-ching
}
} Login Host: 134.192.86.174
} ---------------------------------------------------------------------------
}
} ==============================Original Headers==============================
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} 10, 11 -- Subject: viaWWW: flat embedding mold
} 10, 11 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed"
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}
}


--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff-at-umdnj.edu
**********************************************



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From: oshel1pe-at-cmich.edu
Date: Mon, 7 Jun 2010 10:46:41 -0500
Subject: [Microscopy] TEM of fungi in insect cuticle

Contents Retrieved from Microscopy Listserver Archives
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==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
(Not to me - Oshel)
==========================================================

} Date: Mon, 7 Jun 2010 07:19:26 -0700 (PDT)
} From: Jessica {jegibson-at-syr.edu}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Monday, June 07, 2010
} at 07:19:24 AM.
}
} realname - Jessica
} Email - jegibson-at-syr.edu
} EDUCATION - Graduate College
} QUESTION - Hello,
}
} I study a Family of Fungi (Laboulbeniales) that grow in insect
} integument. I am trying to do TEM analysis with them but I am
} having problems with araldite resin not hardening properly. I have
} made up new resin with new DMP-30 and the resin hardens just fine by
} it self. My sample is at most .3 x .5 x .1 mm. I follow the
} general procedure: after fixation EtOH dehydration using new 100%
} EtOH everytime, 100% proplyene oxide for 15min 2x, 1:4 araldite to
} PO followed by 1:2, 2:1, 4:1 araldite to PO, transfer sample to 100%
} araldite for 1 hr 2x. Then I put them in the molds and into the
} heated pressure chamber. When they come out they are hardened but
} when trying to cut a block face the plastic isn't hard throughout,
} it fractures rather then slices when cut.
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: oshel1pe-at-cmich.edu
Date: Mon, 7 Jun 2010 10:50:37 -0500
Subject: [Microscopy] microscopist salaries in path labs?

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

} Date: Mon, 7 Jun 2010 08:29:45 -0700 (PDT)
} From: Neil Medvitz {nmedvitz-at-bostwicklaboratories.com}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Monday, June 07, 2010
} at 08:29:35 AM.
}
} realname - Neil Medvitz
} Email - nmedvitz-at-bostwicklaboratories.com
} ORGANIZATION - BS Biology
} LOCATION - Glen Allen, VA
} SUBJECT_OF_QUESTION - Anatomic Path Lab Salaries
} QUESTION - In order to make sure my staff members are being paid
} competitively, I would like to ask the List a question regarding
} salaries. I currently run a lab that provides TEM, IHC, IF, histo,
} and enzyme histochemistry, and imaging support for a Anatomical
} Pathology lab. I have trained histo techs in these areas and am
} curious if anyone is willing to offer salary ranges of similar jobs.
} I will do a cost of living adjustment once I receive any info.
} Please respond off line, I will post a summary when I have completed
} collecting information.
}
} Thank you,
} Neil
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: rcommon-at-msu.edu
Date: Mon, 7 Jun 2010 10:58:51 -0500
Subject: [Microscopy] Flat embedding molds

Contents Retrieved from Microscopy Listserver Archives
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I don't think the molds are the problem. Spurr's is very sensitive to
moisture, and if you are using flat molds on a humid day that can result
in poor resin quality. When curing Spurr's, I place the flat molds over
a thin layer of DriRite inside a sealed Tupperware container. The cured
resin quality has been much more consistent since I started using this
procedure.

Ralph Common


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From: david.knecht-at-uconn.edu
Date: Mon, 7 Jun 2010 12:36:24 -0500
Subject: [Microscopy] Excitation/emission profiles of dyes

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

I am wondering about the right way to think about excitation/emission
profiles of dyes. I presume that most of them are done by taking an
absorption spectrum for the excitation spectrum, and a emission
profile by exciting at a specific wavelength (peak?) and scanning
across the emission spectrum. Is that right? The question I have is
whether the absorption spectrum gives you an accurate sense of waht
you will see in emission if you excite at a non-peak wavelength. I
have been surprised numerous times by a strong emission signal coming
from a dye that should not be optimally excited based upon the
absorption spectrum.

Today, I was looking at cells labeled with GFP and mRFP. I was
using a dual filter set and exciting either with either a 480 or 590
LED. The 480 LED is significantly more powerful (5x) than the 590,
but according to the excitation profile of mRFP, you should barely
get any absorption/excitation of mRFP at 480nm. However, I got a
strong RFP signal with the 480LED and very little with the 590 LED.
Obviously this is not quantitative, but I am wondering if it is
misleading to think of the low absorption as an indication that you
will get low emission. Perhaps if you did a total emission energy
with constant excitation power at different excitation wavelengths,
what would it look like and would that be a better way to
characterize dyes?

David Knecht
University of Connecticut
91 N. Eagleville Rd.
Storrs, CT 06268
860-486-2200
david.knecht-at-uconn.edu





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7, 18 -- From: David Knecht {david.knecht-at-uconn.edu}
7, 18 -- Subject: Excitation/emission profiles of dyes
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From: jquetel-at-iwthrowterranova.com
Date: Mon, 7 Jun 2010 18:14:46 -0500
Subject: [Microscopy] viaWWW: looking for journal name

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This Question/Comment was submitted to the Microscopy Listserver
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Email: jquetel-at-iwthrowterranova.com
Name: Julie Quetel

Organization: Withrow & Terranova, Raleigh NC

Title-Subject: [Filtered] looking for journal name

Message: I've got the following citation that I'm trying to locate
today. If anyone has any hints as to where I might find it, even just
the journal name I'd really appreciate it.

Bellare, J.R.,; Thomas, E.L. Proc. Electron Microscopy Soc. Am. 1989, 354.

Login Host: 71.30.86.66
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From: mike.bode-at-resaltatech.com
Date: Mon, 7 Jun 2010 19:51:57 -0500
Subject: [Microscopy] RE: viaWWW: looking for journal name

Contents Retrieved from Microscopy Listserver Archives
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That would be the proceedings of the Electron Microscopy Society meeting
(predecessor to MSA) in San Antonio (47th annual meeting). Not a journal. I
might have that book stashed away somewhere.

---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode at ResAltaTech.com
www.ResAltaTech.com




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From: nizets2-at-yahoo.com
Date: Tue, 8 Jun 2010 03:51:12 -0500
Subject: [Microscopy] viaWWW: Question about Sterility of organisms in Epoxy

Contents Retrieved from Microscopy Listserver Archives
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Hi Robert!

I see at least 3 reasons, which means that 2 are supernumerary:

- Formaldehyde have been long used for the desinfection of surfaces. Perhaps they are still used today in the industry I dont know. If someone argue that your specimen are not surfaces, you can argument that a volume is an infinity of surfaces ;-)
- Dehydration is not compatible with life. Nor is ethanol.
- Temperatures } 60°C for 2 days (for polymerization) are not recommended to maintain life.

And this is not counting with osmication, but I don't know if the sterilizing effect of osmium tetroxide has already been scientifically proven.

Regards,
Stephane


 


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Email: ropope-at-gmail.com
Name: Robert

Title-Subject: [Filtered] Question about Sterility of organisms in Epoxy

Message: Has anyone had dealings with the USDA, CDC, or HHS, or other
agencies with respect to proving sterility of agents that have been
fixed (4%paraformaldehyde/1%gluteraldehyde), osmicated(1% osmium),
(potentially irradiated 2X16[6] rads), dehydrated, and embedded in
epoxy.

I find myself getting questions about things that never were
questioned in the past.

Any and all replies are greatly appreciated.
Robert

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From: nizets2-at-yahoo.com
Date: Tue, 8 Jun 2010 04:12:50 -0500
Subject: [Microscopy] viaWWW: deconvolution & confocal microscopy

Contents Retrieved from Microscopy Listserver Archives
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Hi Molly!

1. If the 2 products you are detecting always colocalize (like DNA and RNA pol II f.ex.), you don't need z-stacks because the probability that you have a colocalization in X-Y plane without colocalization in Z plane in all cells observed is very near from null. But then the less likely the event (colocalization) to occur, the more you'll need to prove that the event really takes place in 3D (meaning, taking Z plane into account). I would suggest that you determine yourself the probability of occurence of the event and determine the need to demonstrate the colocalization in 3D. If needed, just take one representative Z-stack.

2. It depends on the size of your structure and on the thickness of the optical slices. If you have a weak signal, you'll have to open the pinhole and then you increase the size of the optical slice. In this case you'll get improvement from deconvolution. Example: For live cell imaging, decreasing the intensity of the laser (hence the intensity of the fluorescence) is desired, to avoid to affect the cells too much. The best way to know what deconvolution can bring in your personal case is to try it!!

3. I cannot help here sorry.

Hope this helps,

Stephane

 


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Email: mshaw1-at-lumc.edu
Name: Molly Shaw

Organization: Loyola University Chicago

Title-Subject: [Filtered] deconvolution & confocal microscopy

Message: Hello,

I am a first-year graduate student trying to get started with
colocalization analysis of IF images taken by confocal microscopy. I
am working with fixed cells and mainly trying to measure
colocalization of 2 proteins. My main questions are concerning the
use of z-stacks and deconvolution:
1. Are z-stacks always necessary for measurement of colocalization,
or are single images sufficient?
2. I understand that deconvolution is essential for wide-field
images, but does it improve results that much when you're already
using confocal?
3. If you would recommend deconvolution, is the ImageJ plug-in a good
one to use?

Thank you for your time & help. I have read many articles on this
topic, but I keep ending up with conflicting answers.

Thanks again,
Molly Shaw

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From: sedge001-at-umn.edu
Date: Tue, 8 Jun 2010 09:47:33 -0500
Subject: [Microscopy] viaWWW: deconvolution & confocal microscopy

Contents Retrieved from Microscopy Listserver Archives
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Molly,

Scientists have published colocalization results using single planes from
confocal microscopes, z-series, z-series with deconvolution and results
from widefield microscopes with thinly sliced specimens. This alone creates
confusion about the "right" method.

Do yourself a favor and go through the effort of deconvolving images. You
will then have a greater likelihood of being published in a high profile
journal, although you might be asked for more robust methods like
Fluorescence Lifetime Imaging Microscopy (FLIM) or Forster Resonance Energy
Transfer (FRET). Confocal microscopes do collect out of focus fluorescence,
and there is elongation in the z-axis, and deconvolving images ameliorates
those problems.

If nothing else, do the following:

Use a high n.a. lens (1.4 or better).
Mount the specimen in a medium that has a refractive index of 1.4 or better.
Use a coverslip thickness intended for the oil objective (0.17mm).
Make sure you don't just place one image over another (red + green =
yellow--presto! colocalization!); show results using scatterplots.
If the feature you are labeling is at near subresolution dimensions, then
use Nyquist rates for collecting (your core microscopy lab can provide info
about Nyquist rates for their equipment, or at least a way to determine
these).

I have tried Volume J a few years back and it crashed with large image
sets. I suspect computers/software have caught up by now. Certainly Image
J/Volume J is good for attempting deconvolution. It's advisable to make a
point spread function (psf) from subresolution beads (0.8 microns, or what
you read in the literature as the bead to use for the objective: I can't
remember offhand what I used way back when) and then deconvolve using the
psf.

Have fun!

Jerry Sedgewick

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From: jfmjfm-at-umich.edu
Date: Tue, 8 Jun 2010 10:26:32 -0500
Subject: [Microscopy] Posting for Peter Crozier at ASU

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Please reply to Peter Crozier if you have questions.

Opening: Post-Doctoral Research Associate On Electron Energy-Loss Spectroscopy of Carbonaceous Aerosols

A post-doctoral research associate position is available to work on the application of electron energy-loss spectroscopy (EELS) to atmospheric aerosols. The primary research focus will be the development and application of electron energy-loss spectroscopy to problems in atmospheric science related to aerosols and global climate change. The successful candidate will use atomic resolution imaging and electron energy-loss spectroscopy to determine the optical properties of carbonaceous aerosols. Candidates should have a Ph.D. in Physics, Materials Science or related disciplines. The position requires extensive experience in transmission electron microscopy and electron energy-loss spectroscopy. Experience working with low-loss EELS and monochromators is also preferred. This provides an opportunity to work on cutting-edge research related to climate change. ASU is a leader in the field of transmission electron microscopy and part of this work will be performed on state-of-the-art aberration corrected, monochromated instruments.

This postdoctoral position is available immediately, will be renewable on an annual basis, and is anticipated for at least two years. Compensation will be commensurate with qualification and experience. Health benefits will also be provided. The position is open until it is filled.

Interested candidates can apply by email by sending a cover letter, CV with a complete list of publications, and contact information of 3 professional references to: Prof. Peter A. Crozier, School of Mechanical, Aerospace, Chemical and Materials Engineering, Arizona State University, P.O. Box 876106, Tempe, AZ 85287-6106, email crozier-at-asu.edu.
--
John Mansfield PhD Cphys MInstP
2010 Microscopy & Microanalysis Program Chair
North Campus Electron Microbeam Analysis Laboratory
417 SRB, University of Michigan
2455 Hayward, Ann Arbor MI 48109-2143 USA
Phone: (734) 936-3352 FAX (734) 763-2282
Cell. Phone: (734) 834-3913
(Leaving a phone message at 936-3352 is preferable to 834-3913)
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From: maryflet-at-interchange.ubc.ca
Date: Tue, 8 Jun 2010 10:38:05 -0500
Subject: [Microscopy] viaWWW: looking for journal name

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Dear Julie,
As Mike bode said, that was the Proceedings of the 1989 EMSA meeting. The
Proceedings published two-page abstracts of the papers presented at the
meeting.
I have the volume. Would you like to have a PDF of the pages?
Regards,

Mary Fletcher
Electron Microscopist
Materials Eng. UBC
#309 - 6350 Stores Road
Vancouver, B.C. V6T 1Z4
Canada
Tel: 604-822-5648
Fax: 604-822-3619
email: maryflet-at-interchange.ubc.ca


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Organization: Withrow & Terranova, Raleigh NC

Title-Subject: [Filtered] looking for journal name

Message: I've got the following citation that I'm trying to locate
today. If anyone has any hints as to where I might find it, even just
the journal name I'd really appreciate it.

Bellare, J.R.,; Thomas, E.L. Proc. Electron Microscopy Soc. Am. 1989, 354.

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From: klivi-at-jhu.edu
Date: Tue, 8 Jun 2010 12:27:29 -0500
Subject: [Microscopy] used XP3 processor

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We are in need of a used Oxford XP3 pulse processor to temporarily
replace our dead processor on an old Philips 420. Repair of this unit
can be expensive and we need to tide ourselves over till a new scope/
analytical system has been acquired. Please, no solicitations for new
units.
Thanks
Ken


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From: lebeau-at-mrl.ucsb.edu
Date: Tue, 8 Jun 2010 13:10:26 -0500
Subject: [Microscopy] Post-doc Position - Posting for Susanne Stemmer at UCSB

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Please reply to Susanne Stemmer if you have questions.

Opening: Postdoctoral Position in Transmission Electron Microscopy at the University of California, Santa Barbara

Projects include diffraction-contrast TEM and scanning transmission electron microscopy studies of interfaces, defects and microstructures in oxide and semiconductor thin films grown by molecular beam epitaxy. Applicants should have demonstrated expertise in problem solving in materials science using a wide range of transmission electron microscopy techniques. Facilities at UCSB include a FEI Titan TEM/STEM, several LaB6 TEMs, FIB, Imago LEAP, etc. The position is available starting July 2010. Duration is about 1-3 years and salary is commensurate with qualifications.

Interested candidates should send their resume and the names and contact information of two to three references to:

Professor Susanne Stemmer
Materials Department
University of California
Santa Barbara, CA 93106-5050
Phone: (805) 893-6128
Fax: (805) 893-8486
stemmer-at-mrl.ucsb.edu
http://www.mrl.ucsb.edu/~stemmer


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From: Bplowman-at-pacific.edu
Date: Tue, 8 Jun 2010 21:43:04 -0500
Subject: [Microscopy] viaWWW: Photography with a dissecting microsope

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Email: Bplowman-at-pacific.edu
Name: Barbara L. Plowman

Organization: Univ. of the Pacific AADugoni School of Dentistry

Title-Subject: [Filtered] Photography with a dissecting microsope

Message: I would like to know which adapter is used with a
camera(digital or otherwise)on a dissecting microscope.

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6, 11 -- To: microscopy-at-microscopy.com
6, 11 -- From: Bplowman-at-pacific.edu (by way of MicroscopyListserver)
6, 11 -- Subject: viaWWW: Photography with a dissecting microsope
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From: seawall90-at-gmail.com
Date: Tue, 8 Jun 2010 21:43:43 -0500
Subject: [Microscopy] viaWWW: Old Parts - Noran Tracor Unix Based Computer

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Email: seawall90-at-gmail.com
Name: Sean Wallace

Organization: Ambient Group, Inc

Title-Subject: [Filtered] Old Parts - Noran Tracor Unix Based Computer

Message: Our old computer has given up the ghost. Does anyone have
any insight into such obsolete technology? Noran system dectector
attached to our Hitachi 600AB TEM

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==============================Original Headers==============================
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6, 11 -- Subject: viaWWW: Old Parts - Noran Tracor Unix Based Computer
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From: lydiaj-at-stanford.edu
Date: Tue, 8 Jun 2010 21:44:31 -0500
Subject: [Microscopy] viaWWW: SEM: cleaning of BSE detector

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Email: lydiaj-at-stanford.edu
Name: Lydia Joubert

Organization: CSIF, Stanford University

Title-Subject: [Filtered] SEM: cleaning of BSE detector

Message: Dear Microscopists

The BSE detector in our Hitachi Variable-Pressure SEM has accumulated
excessive dirt from the spattering of wet biomaterials at short WD,
and this interferes greatly with the quality of detection and imaging.

Any suggestions on cleaning this BSE detector? It is a 5 Quadrant
Silicon chip, and will not tolerate physical wiping. The manufacturer
does not guarantee cleaning operations. If anyone who has already
cleaned this delicate tool successfully can let me know their secret,
it will be greatly appreciated!

Thanks
Lydia


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From: steven.samuelsson-at-sri.com
Date: Tue, 8 Jun 2010 21:44:53 -0500
Subject: [Microscopy] viaWWW: Freeze-Fracture Electron Microscopy

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Email: steven.samuelsson-at-sri.com
Name: Steve Samuelsson

Organization: SRI International, Inc

Title-Subject: [Filtered] Freeze-Fracture Electron Microscopy

Message: Listservers: am interested to know who is actively using FF
in their research or as a service and to what level of application.

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From: vladislav_speransky-at-nih.gov
Date: Tue, 8 Jun 2010 22:26:35 -0500
Subject: [Microscopy] Re: viaWWW: Question about Sterility of organisms in Epoxy

Contents Retrieved from Microscopy Listserver Archives
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Well, I see noone has responded from CDC...
Officially, there are certain hard rules for dealing with really bad
pathogens.
A few years ago, we had a dinner talk at our local Chesapeake Society
for Microscopy. Sorry I forgot the guy's name, but he was describing
his work doing EM of stuff like Ebola virus. That included thin
section EM, tissues of those sick monkeys here in Virginia. The
quality of preservation was pretty bad. And the reason for that?
Because, he said, they had to fix the samples according to certain
very strict rules made by CDC. The protocol prescribed started with
many hours (2 days?) in FA, no GA. Clearly, whoever made those rules,
must have not been primarily an EM person...

-at- OP:
It would help if you were more specific - what kind of "agents" are
you talking about?
One very famous example, mentioned a few times by Sara Miller when a
similar question would pop up, is mammalian prions. That stuff seems
to be infectious even after having been under EM, if I remember right!..

________________________________________________
Vlad Speransky, Staff Scientist
Supramolecular Structure and Function Resource
National Institute of Biomedical Imaging and Bioengineering, NIH
13 South Dr, Rm. 3N17 MSC 5766
Bethesda, MD 20892
301 496-3989
vladislav_speransky-at-nih.gov
http://www.nibib.nih.gov/Research/Intramural/lbps/staff/speransky

Opinions and experiences related are those of Vlad Speransky and do
not represent the NIH. On the good side, this message is not
confidential and can be freely shared and reproduced.

} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi Robert!
}
} I see at least 3 reasons, which means that 2 are supernumerary:
}
} - Formaldehyde have been long used for the desinfection of surfaces.
} Perhaps they are still used today in the industry I dont know. If
} someone argue that your specimen are not surfaces, you can argument
} that a volume is an infinity of surfaces ;-)
} - Dehydration is not compatible with life. Nor is ethanol.
} - Temperatures } 60°C for 2 days (for polymerization) are not
} recommended to maintain life.
}
} And this is not counting with osmication, but I don't know if the
} sterilizing effect of osmium tetroxide has already been
} scientifically proven.
}
} Regards,
} Stephane
}
}
}
}
}
} ----- Original Message ----
} X-from: "ropope-at-gmail.com" {ropope-at-gmail.com}
} To: nizets2-at-yahoo.com
} Sent: Thu, June 3, 2010 2:39:48 AM
} Subject: [Microscopy] viaWWW: Question about Sterility of organisms
} in Epoxy
}
}
}
}
} ----------------------------------------------------------------------------
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} America
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} This Question/Comment was submitted to the Microscopy Listserver
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} please copy both ropope-at-gmail.com as well as the MIcroscopy
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}
} Email: ropope-at-gmail.com
} Name: Robert
}
} Title-Subject: [Filtered] Question about Sterility of organisms in
} Epoxy
}
} Message: Has anyone had dealings with the USDA, CDC, or HHS, or other
} agencies with respect to proving sterility of agents that have been
} fixed (4%paraformaldehyde/1%gluteraldehyde), osmicated(1% osmium),
} (potentially irradiated 2X16[6] rads), dehydrated, and embedded in
} epoxy.
}
} I find myself getting questions about things that never were
} questioned in the past.
}
} Any and all replies are greatly appreciated.
} Robert


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From: vladislav_speransky-at-nih.gov
Date: Tue, 8 Jun 2010 22:34:38 -0500
Subject: [Microscopy] pdf publication policies

Contents Retrieved from Microscopy Listserver Archives
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Stephane, I would just email the request. Then one more time, if they
don't respond ;)
Unless the person is totally jaded/misanthropic, this should be
pleasantly flattering to them!

Good old days...
I still cherish a reprint request I got in early 90s from Bjorn
Afzelius. Will frame it one day...

________________________________________________
Vlad Speransky, Staff Scientist
Supramolecular Structure and Function Resource
National Institute of Biomedical Imaging and Bioengineering, NIH
13 South Dr, Rm. 3N17 MSC 5766
Bethesda, MD 20892
301 496-3989
vladislav_speransky-at-nih.gov
http://www.nibib.nih.gov/Research/Intramural/lbps/staff/speransky

Opinions and experiences related are those of Vlad Speransky and do
not represent the NIH. On the good side, this message is not
confidential and can be freely shared and reproduced.

} ----------------------------------------------------------------------------
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}
} I've had requests for pdfs of papers, and am happy to send them, and
} have
} found others happy to email copies of their papers. For example, our
} library cannot afford to pay for immediate online access to all
} journals,
} and if I want a pdf of a very recent article, they have to photocopy
} it and
} send that, they cannot send the high quality pdf until the waiting
} time -
} 6-12 months - is over. So if it's an article in which good quality
} image
} reproduction is essential, I usually email the author. I could wait
} for 6
} months....but by then I'd have forgotten why I wanted to read it.
} cheers,
} Rosemary
}
} Dr Rosemary White
} CSIRO Plant Industry
} GPO Box 1600
} Canberra, ACT 2601
}
} T 02 6246 5475
} F 02 6246 5334
} E rosemary.white-at-csiro.au
}
} On 2/06/10 12:47 AM, "Rob.Bowen-at-caddock.com" {Rob.Bowen-at-caddock.com}
} wrote:
}
} }
} }
} }
} }
} ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} }
} ----------------------------------------------------------------------------
} }
} } Hi, Stephane,
} } I've been grumbling about the $30 for a pdf, too. However, I
} realized
} } that the big money is if you want the article RIGHT NOW!!!! If I
} want to
} } get up from my chair, trundle down to the local library and fill
} out a paper
} } form with the citation info, and pay an interlibrary loan fee of
} $2 or $3,
} } and wait several days, I can get an adequate quality copy from
} someplace
} } that has the publication. The. Inertia still keeps me from getting
} a lot of
} } articles this way, but I figure having some tax money go to the
} library is
} } definitely a bet buy.
} }
} } Rob Bowen
} }
} } } From: {nizets2-at-yahoo.com}
} } } Reply-To: {nizets2-at-yahoo.com}
} } } Date: Tue, 1 Jun 2010 02:19:24 -0500
} } } To: {rob.bowen-at-caddock.com}
} } } Subject: [Microscopy] pdf publication policies
} } }
} } }
} } }
} } }
} } }
} ----------------------------------------------------------------------------
} } } The Microscopy ListServer -- CoSponsor: The Microscopy Society
} of America
} } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } } On-Line Help http://www.microscopy.com/MicroscopyListserver/
} FAQ.html
} } }
} ----------------------------------------------------------------------------
} } }
} } } Hi to all,
} } }
} } } I am sorry if this message does not talk about microscopy but I
} have a
} } } question about the publication and duplication policies of
} scientific papers.
} } } When I was student (in my head it was yesterday but it may be a
} little bit
} } } earlier), before the pdf era, when we wanted a paper we had
} preprinted cards
} } } that we sent to the authors and there was a kind of unwritten law
} that wanted
} } } that the authors graciously send a copy of their paper (however
} if they did
} } } not, you couldn't sue them ;-)).
} } } Now the publications are online and if we want a paper we have to
} graciously
} } } pay something between 30 and 50 dollars.
} } } But what about the good old unwritten law? Is it still possible
} to send an
} } } email to an author and ask for a pdf copy of his paper or do we
} really have
} } } to
} } } translate everything down on earth in $$?
} } }
} } } Regards,
} } }
} } } Stephane
} } }

==============================Original Headers==============================
5, 23 -- From vladislav_speransky-at-nih.gov Tue Jun 8 22:34:38 2010
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From: schooley-at-mcn.org
Date: Tue, 8 Jun 2010 23:00:38 -0500
Subject: [Microscopy] Re: viaWWW: Photography with a dissecting microsope

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

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

Barbara -

You'll find a simple systems described on Project MICRO's website at
http://www.microscopy.org/education/projectMICRO/camera.cfm or
http://www.einstein.yu.edu/aif/gallery/cheapscope/index.htm

You can also use a short length of tubular black plastic pipe
insulating foam from the hardware store to couple the camera lens to
the eyepiece. Mount the camera on a separate stand to avoid
vibration.

Caroline
--
Caroline Schooley
Project MICRO Coordinator
Microscopy Society of America
45301 Caspar Point Road, Box 117
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.microscopy.org/ProjectMICRO

==============================Original Headers==============================
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5, 18 -- To: Bplowman-at-pacific.edu
5, 18 -- From: Caroline Schooley {schooley-at-mcn.org}
5, 18 -- Subject: Re: [Microscopy] viaWWW: Photography with a dissecting microsope
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From: naomi_mccallum-at-health.qld.gov.au
Date: Wed, 9 Jun 2010 00:11:46 -0500
Subject: [Microscopy] Fwd: Re: viaWWW: Photography with a dissecting microsope

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

Hi Barbara

Unfortunately you have not mentioned the model of the Dissecting Microscope but perhaps the local agent for that brand would be the first point of contact. If they don't stock what you require then perhaps they might have a detailed description. Then you might be lucky enough to find someone that has finished with theirs and happy to help you out.

Kind regards
Naomi

} } } {schooley-at-mcn.org} 9/06/2010 2:07 pm } } }



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

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

Barbara -

You'll find a simple systems described on Project MICRO's website at
http://www.microscopy.org/education/projectMICRO/camera.cfm or
http://www.einstein.yu.edu/aif/gallery/cheapscope/index.htm

You can also use a short length of tubular black plastic pipe
insulating foam from the hardware store to couple the camera lens to
the eyepiece. Mount the camera on a separate stand to avoid
vibration.

Caroline
--
Caroline Schooley
Project MICRO Coordinator
Microscopy Society of America
45301 Caspar Point Road, Box 117
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.microscopy.org/ProjectMICRO

==============================Original Headers==============================
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5, 18 -- To: Bplowman-at-pacific.edu
5, 18 -- From: Caroline Schooley {schooley-at-mcn.org}
5, 18 -- Subject: Re: [Microscopy] viaWWW: Photography with a dissecting microsope
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From: sergei2-at-ornl.gov
Date: Wed, 9 Jun 2010 10:35:43 -0500
Subject: [Microscopy] SPM for Energy Applications - ORNL September 15

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

Dear colleagues

I would like to attract your attention to the 1st Workshop on Scanning
Probe Microscopy for Energy Applications, to be held at the Center for
Nanophase Material Sciences at Oak Ridge National Laboratory, and
jointly organized by CNMS, FIRST center, and Asylum Research, on
September 15-17, 2010. The workshop features a series of keynote and
invited talks that cover recent advances in characterization of energy
materials systems using Scanning Probe Microscopy (SPM) techniques, as
well as the state of the art in energy dissipation and transformation
measurements by SPM. The two-day meeting will also include contributed
talks and a poster session. An equipment lab will be held on the last
day for demonstration of recently-developed dynamic and multi-spectral
SPM modes and hands-on tutorials. Ultimately, the workshop goal is to
build a network of materials scientists centered on the applications of
SPM for energy problems and to promote rapid dissemination of
theoretical knowledge, experimental protocols, and novel technique
development in this rapidly growing area.

The detailed information on the workshop, as well as abstract submission
and pre-registration form are available at
http://www.asylumresearch.com/Energy/

Yours

Sergei V. Kalinin

--
Sergei V. Kalinin
co-Theme Leader for Functional Imaging on the Nanoscale
The Center for Nanophase Materials Sciences
and Materials Sciences and Technology Division
Oak Ridge National Laboratory
Oak Ridge, TN 37922

Section editor, Nanotechnology

Adjunct Associate Professor,
Department of Materials Science and Engineering,
University of Tennessee, Knoxville

Phone: (865) 241-0236
http://imaging.ornl.gov


==============================Original Headers==============================
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From: Phil.Ahrenkiel-at-sdsmt.edu
Date: Wed, 9 Jun 2010 14:56:24 -0500
Subject: RE: probe for CBED with CA out

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

I got two interesting responses to my query about forming a small probe with a large convergence angle on an H-7000 FA TEM (shown below). It seems we found a reasonable solution on our own: Raise the sample above the eucentric height, refocus the objective lens (and make a few other adjustments), then focus the condenser lens to form a convergent probe. The convergence angle increases with increasing height. Above a certain height, the convergence angle (keeping the CA in) turns out to be even bigger than it is at the eucentric height with the CA out, but without all the distortion that prevented us from forming a small probe.

Two problems: the sample is not eucentric, and the camera length seems to be about 30% smaller than usual, but those things we can deal with.

Thanks for the comments and suggestions.

-Phil
------------------------------------------
Phil Ahrenkiel, Assistant Professor
Nanoscience and Nanoengineering Ph.D. Program
South Dakota School of Mines and Technology
501 E. Saint Joseph St.
Rapid City, SD 57701 U.S.A.
Office: EP 221
Phone: 605-394-5238, Fax: 605-394-2365
Email: Phil.Ahrenkiel-at-sdsmt.edu


-----Original Message-----
X-from: Qingfeng Xing [mailto:qxing-at-ameslab.gov]
Sent: Saturday, June 05, 2010 7:48 AM
To: Ahrenkiel, Phil

Hello-

A student in my group is using CBED on our Hitachi H-7000 FA TEM. One thing though: we usually take the condenser aperture out to get a large convergence angle for measuring HOLZ lines. But now we are noticing that we can't form a probe smaller than about 500 nm with the CA out, because of a coma-type of distortion that affects the probe shape, so it doesn't form a circular spot. In fact, the problem seems even worse in Fine-Probe mode (which turns the middle condenser lens on). The probe looks fine with the CA in, but the convergence semi-angle is only about 3 mrad, whereas it is 8 mrad with the CA out. Does anyone you know if there is an alignment procedure for an H-7000 that can reduce this problem (e.g., coma-free alignment), or is it just an inherent artifact? I'd like to buy a CA strip with some bigger holes than what we have, but it's hard to know in advance if that would fix this. Thanks for any input.

-Phil
------------------------------------------
Phil Ahrenkiel, Assistant Professor
Nanoscience and Nanoengineering Ph.D. Program
South Dakota School of Mines and Technology
501 E. Saint Joseph St.
Rapid City, SD 57701 U.S.A.
Office: EP 221
Phone: 605-394-5238, Fax: 605-394-2365
Email: Phil.Ahrenkiel-at-sdsmt.edu





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From: Gregory.Hendricks-at-umassmed.edu
Date: Wed, 9 Jun 2010 20:54:39 -0500
Subject: [Microscopy] viaWWW: rotation grid holder

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Email: Gregory.Hendricks-at-umassmed.edu
Name: Greg Hendricks

Organization: UMass Medical School

Title-Subject: [Filtered] rotation grid holder

Message: Does anyone have a rotation holder for an Philips CM120 or
FEI Tecnai BioTwin that they would be willing to donate, lend for a
few months, or sell second hand? Any help greatly appreciated.
Thanks.

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From: mweschler-at-hotmail.com
Date: Wed, 9 Jun 2010 20:55:03 -0500
Subject: [Microscopy] viaWWW: Wanted: JEOL 1010 TEM

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Email: mweschler-at-hotmail.com
Name: Matt Weschler

Title-Subject: [Filtered] Wanted: JEOL 1010 TEM

Message: I am in need of a JEOL 1010 TEM. If anyone has one they
would like to disposition, please contact me.

Thank you,
Matt Weschler


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From: J.Nailon-at-uq.edu.au
Date: Wed, 9 Jun 2010 20:55:24 -0500
Subject: [Microscopy] [Filtered] MicroscopyListserverviaWWW: Position Vacant

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Email: J.Nailon-at-uq.edu.au
Name: John Nailon

Organization: University Of Queensland

Title-Subject: [Filtered] Position Vacant

Message: The Centre for Microscopy and Microanalysis (CMM) at the
University of Queensland are searching for an expert Cryo-TEM
Scientist/Engineer to manage its Cryo-TEM Facility. The CMM operates
five laboratories across the St.Lucia, Brisbane, Campus. The Cryo-TEM
Facility is equipped with 3 TEMs, the flagship being an FEI T30F
Cryo-TEM.
For the Position Description and Selection Criteria please email me,
applications close soon.

Regards
John V Nailon
Operations Manager
University of Queensland
St.Lucia QLD 4072 Australia

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From: kjmorris-at-well.ox.ac.uk
Date: Thu, 10 Jun 2010 04:00:07 -0500
Subject: [Microscopy] viaWWW: deconvolution & confocal microscopy

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Hi Molly

Just to add to Stephane's and Jerry's comments. You are right that a
confocal will not benefit from deconvolution software anywhere near as much
as a widefield microscope, for the reason you suggest, i.e. much of the
out-of focus information is removed by the confocal iris/pinhole in any
case. I use Improvison's [Perkin Elmer] 3D Volocity software to look at
things like 3D images, 3D cellular volumes and cell morphometry. In order to
get the best from a Z slice you are advised to over sample the z-stack, i.e.
for a 0.9um optical slice capture the z stack at least 0.45um apart [our
Zeiss 510 has a button to automatically 'double oversample' like this]. For
the likes of Volocity deconvolution, Improvision recommended to me that you
massively oversample the confocal z-stack with ten or more slices to every
0.9um z focus step [I think the rep suggested perhaps up to as high as 100x
oversampling]. I rarely use the optional 'Restoration' module
[deconvolution] Volocity module as I only use the confocal for z-stack, and
double oversampling and 3D reconstruction alone with Volocity is adequate
for my needs.

For co-localisation I would agree with Stephane though, and initially try
and tackle the co-localisation quantification in 2D using MetaMorph
[probably just because it's just easier]. I don't use ImageJ very much as I
have a MetaMorph licence, and we don't have the 3D Quantitation module for
Volocity in any case, that might be needed for 3D co-localisation
quantification - if interested just ask Improvision about that as their
support is superb.

Always be suspicious that any co-localisation isn't bleedthrough. You can
check this via spectral un-mixing on a 510 Metahead or just try imaging
single labeled samples of each fluorochrome and check there's no signal in
the other channels where there is now no longer any label [at the same
confocal imaging settings]. Don’t' forget a sample with no label just in
case of autofluorescence.

To quote Improvisions own website: "If you acquire your images from a
widefield microscope and want to volume render or make measurements from the
image data, you will need to use an image restoration technique
[de-convolution] to remove the out-of-focus information - a product of the
optical properties of the microscope. Even if your images are captured using
a laser scanning or spinning disk confocal microscope, the image quality may
benefit from image restoration." The word 'may' is probably significant.

Regards

Keith
---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford  OX3 7BN,
United Kingdom.

Telephone:  +44 (0)1865 287568
Email:  kjmorris-at-well.ox.ac.uk
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy

-----Original Message-----
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Sent: 07 June 2010 14:37
To: kjmorris-at-well.ox.ac.uk

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Email: mshaw1-at-lumc.edu
Name: Molly Shaw

Organization: Loyola University Chicago

Title-Subject: [Filtered] deconvolution & confocal microscopy

Message: Hello,

I am a first-year graduate student trying to get started with
colocalization analysis of IF images taken by confocal microscopy. I
am working with fixed cells and mainly trying to measure
colocalization of 2 proteins. My main questions are concerning the
use of z-stacks and deconvolution:
1. Are z-stacks always necessary for measurement of colocalization,
or are single images sufficient?
2. I understand that deconvolution is essential for wide-field
images, but does it improve results that much when you're already
using confocal?
3. If you would recommend deconvolution, is the ImageJ plug-in a good
one to use?

Thank you for your time & help. I have read many articles on this
topic, but I keep ending up with conflicting answers.

Thanks again,
Molly Shaw

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From: oshel1pe-at-cmich.edu
Date: Thu, 10 Jun 2010 08:48:43 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist

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==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member of MSA,
and any reply should go directly to the poster as well as to the list.
==========================================================

} Date: Sun, 6 Jun 2010 20:34:53 -0700 (PDT)
} From: Joey {yeojoey1986-at-gmail.com}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Sunday, June 06, 2010
} at 08:34:50 PM.
}
} realname - Joey
} Email - yeojoey1986-at-gmail.com
} ORGANIZATION - University of Melbourne
} LOCATION - Melbourne, Victoria, Australia
} SUBJECT_OF_QUESTION - What is optical diffraction limit?
} QUESTION - Hi, I would like to ask what is the exact meaning of
} optical diffraction limit, especially for the case of an IR
} microscope?
}
} THanks!
}
} Joey
}

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From: sarj0007-at-unf.edu
Date: Thu, 10 Jun 2010 11:13:43 -0500
Subject: [Microscopy] Ultra microtome manual

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I was wondering if anyone had an electronic copy of the manual for the Serval Porer-Blum MT-2, or know where to track one down. We have dragged ours out of the closet and want to double-check some of the knobs are working like they should be. Thanks.

-Jason Saredy


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From: p.ingram-at-cellbio.duke.edu
Date: Thu, 10 Jun 2010 16:28:57 -0500
Subject: [Microscopy] Give-away!

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To all Listers,

We are currently housekeeping our lab and would like to give
away the following:

An AGFA DUOSCAN T2500 high resolution scanner. It comes with
all cables, connectors, various sized negative, slide and sheet
holders together with full instruction manuals & documentation; it is
in immaculate condition. The software is for Apple Macintosh
computers (through OS 9 machines equipped with Nubus), although I
believe you can still download Mac OS X and Windows software from the
Agfa website
(see
http://static.agfa.com/digicam_scanner_drivers/scanner/duoscan/index.html)

If you are interested, please contact me off-line. All you
will have to pay for is shipping costs.

If we get no takers by June 25th. this fine instrument will
have to be scrapped.

Peter


--
Peter Ingram
Sr. Physicist
Adj. Professor of Pathology
Duke University Medical Center
PO Box 90319
DURHAM NC 27708-0319

Tel: 919 660-2695
Fax: 919 660-2671
Email: p.ingram-at-cellbio.duke.edu

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From: larry.ackerman-at-ucsf.edu
Date: Thu, 10 Jun 2010 17:42:57 -0500
Subject: [Microscopy] oxidised grids

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Hi,
I am using up a large stock of inherited grids from various vendors and
discovered that many of the copper grids develop a dark black surface
after coating with a support film. I have used Butvar and Ultem
(polyetherimide also known as PEI) with chloroform as the solvent. I
tried lightly etching with 1N HCl and always clean with ethanol. The
"oxidation" continues and after a month or so the grids have "hairy"
edges and the films are contaminated. I have never seen this previously
and it has not happened with nickel grids.

Any suggestions for eliminating the "oxidation?"

Thanks
Larry
--
Larry Ackerman, Specialist
Electron Microscopy Lab
Manager, DERC Microscopy Core
UCSF, Dept. of Anatomy, Rm S1355
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

415-999-4758

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From: drg.mitchell-at-sydney.edu.au
Date: Thu, 10 Jun 2010 21:08:18 -0500
Subject: [Microscopy] Re: oxidised grids

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Dear Larry

Re: [Microscopy] oxidised grids

A soak in some dilute orthophosphoric acid will generally clean up tarnished
copper. A final rinse with ethanol should then see them clean and ready to
go.

Regards

Dave Mitchell


Dr David Mitchell
Senior Microscopist, Laboratory Manager (Acting)

Contact:
PH +61 2 9036 7633
FAX +61 2 9351 7682
drg.mitchell-at-sydney.edu.au

Address:
Australian Centre for Microscopy & Microanalysis
Incorporating:
Australian Microscopy & Microanalysis Research Facility
ARC Centre of Excellence for Design in Light Metals
Madsen Building F09, Room 128A
The University of Sydney
NSW, 2006, Australia
sydney.edu.au/acmm {http://www.sydney.edu.au/acmm}
www.ammrf.org.au {http://www.ammrf.org.au/}

www.dmscripting.com




On 11/6/10 8:51 AM, "larry.ackerman-at-ucsf.edu" {larry.ackerman-at-ucsf.edu}
wrote:

}
}
}
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} Hi,
} I am using up a large stock of inherited grids from various vendors and
} discovered that many of the copper grids develop a dark black surface
} after coating with a support film. I have used Butvar and Ultem
} (polyetherimide also known as PEI) with chloroform as the solvent. I
} tried lightly etching with 1N HCl and always clean with ethanol. The
} "oxidation" continues and after a month or so the grids have "hairy"
} edges and the films are contaminated. I have never seen this previously
} and it has not happened with nickel grids.
}
} Any suggestions for eliminating the "oxidation?"
}
} Thanks
} Larry


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From: maevat-at-udel.edu
Date: Thu, 10 Jun 2010 21:33:27 -0500
Subject: [Microscopy] viaWWW: Preferential staining of PMMA with iodine or other

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Email: maevat-at-udel.edu
Name: Maeva

Organization: University of Delaware

Title-Subject: [Filtered] Preferential staining of PMMA with iodine
or other staining agents

Message: Hello,
I have a poly(ethylene-alt-propylene-b-styrene-b-methyl methacrylate)
[PEP-PS-PMMA] triblock copolymer system where the PEP is in fact a
partially hydrogenated poly(isoprene) at 90%. I have seen in
literature that PMMA can be preferentially stained by iodine in a
PS-PEP-PMMA system. The paper does not give much details on the
iodine staining procedure. I was wondering if anybody have stained
PMMA with iodine or any other staining agent (one that do not stain
the other two blocks) and could give me some tips on how to go about
that?


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From: dmywong-at-ucdavis.edu
Date: Thu, 10 Jun 2010 21:33:49 -0500
Subject: [Microscopy] viaWWW: Algae Cell Fixation

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Email: dmywong-at-ucdavis.edu
Name: Diana Wong

Organization: Chemistry

Title-Subject: [Filtered] Algae Cell Fixation

Message: I would like the view my algae cells with a confocal
microscope. The cell sizes are 1 um and 12 um.

I am interested in seeing:
1.) Cell surface with lipid droplet fluorescing (using BODIPY 505/515
and NIle Red dye)
2.) Cell surface

What is the best method to fix them on a slide?

Thanks you for your help,
Diana

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From: drg.mitchell-at-sydney.edu.au
Date: Thu, 10 Jun 2010 22:23:19 -0500
Subject: [Microscopy] TEM: CCD camera dark field response

Contents Retrieved from Microscopy Listserver Archives
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Dear Listers

I have a query about the dark response of my CCD (Gatan, Ultrascan 2k x 2k).
The camera is superb and I am certainly not criticising it in any way. The
effects I am describing are totally insignificant for routine imaging. The
only reason I found them was because I am doing radial distribution function
work, and even tiny dark offsets are very evident in my results.

What I did:
Camera at stable temperature for many weeks.
No intense illumination on the camera for 12+ hours
Beam off, and no room light entering the column (2200FS TEM)
Gain Calibration performed
Removed all pre-existing dark references.

I then captured a series of images (2k x 2k) - binning 1x for an integrated
exposure of 100s (different numbers of frames at different exposures). These
were gain and dark reference corrected. The total counts in the images were
then summed. The results are shown below:

Frames Exposure/s Sum of counts

200 0.5 4.40E+07
100 1 1.66E+07
50 2 1.19E+07
20 5 8.98E+05
10 10 -8.95E+05
4 25 -5.79E+05
1 100 -1.23E+05



Clearly the dark counts increase significantly as the number of frames
increases. This suggests that there is some kind of readout noise which is
non-random and is always positive. The mean and standard deviation values
for the above scaled in a similar manner to the sum value.

In the second experiment I set up my camera to acquire frames constantly in
view mode using a short exposure 0.0125s, 4x binning. Periodically, I
captured a 1s frame by hitting acquire. This would cause the view mode to
pause, the frame to be acquired and then the view mode would start again
immediately. I repeated this several times and the summed dark counts I
obtained for each image are shown below:

213119
343479
149075
230461

Average=236783
Std Dev=85916


I then stopped the view mode running and acquired single frames every 30s
with a 1s exposure. The summed dark counts I obtained were:

522476
480510
584586

Average=529190
Std Dev=52361

I then started View mode running again and periodically acquired 1s images
as I did earlier. The summed dark counts were

299767
279014

Average=289390
Std Dev=14674s

The difference in the dark counts between the two modes is quite
reproducible.

The conclusion from the above is that if one acquires an image immediately
after the camera has been very recently accessed (in View mode), it produces
fewer dark counts than if it has been idling for a while.

These observations suggest that there may be two separate phenomena at work.
In the first case a constant readout noise accompanies each acquisition,
whereas in the second the act of stripping charge from a CCD results in a
short-lived reduction in the noise the camera produces.

If anyone has any comments on this, I'd be interested. From an experimental
viewpoint none of this is critical. It is of no significance in routine
imaging, and in RDF analysis I can simply correct any offsets by adding a
constant. The motivation for doing this analysis was to understand the
physical reason for needing the constant.

Thanks and regards,

Dave Mitchell


Dr David Mitchell
Senior Microscopist, Laboratory Manager (Acting)

Contact:
PH +61 2 9036 7633
FAX +61 2 9351 7682
drg.mitchell-at-sydney.edu.au

Address:
Australian Centre for Microscopy & Microanalysis
Incorporating:
Australian Microscopy & Microanalysis Research Facility
ARC Centre of Excellence for Design in Light Metals
Madsen Building F09, Room 128A
The University of Sydney
NSW, 2006, Australia
sydney.edu.au/acmm {http://www.sydney.edu.au/acmm}
www.ammrf.org.au {http://www.ammrf.org.au/}

www.dmscripting.com



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From: nizets2-at-yahoo.com
Date: Fri, 11 Jun 2010 03:14:19 -0500
Subject: [Microscopy] viaWWW: deconvolution & confocal microscopy

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

A word of caution about the part of the message from Keith dealing with quantification.
Be VERY careful with quantification in fluorescence. A higher signal doesn't necessarily mean a higher concentration of the antigen, it may also mean a better antibody, a better accessibility for the antibody, a better stability from the laser light and so many other things.
You could trace a line through the structure and show a profile scan (confocals are good at that), this is a nice way to concentrate the attention only on one structure and demonstrate the colocalization of 2 peaks of intensity. The profile also allows a fine analysis of the both signals intensities and who knows, perhaps you'll discover that both signals are not just perfectly superimposed, but are next to each other (very close), or that one signal has more a ring pattern and so on. Small differences in intensities are not easily appreciated with eyes (especially at high intensities), but when they are presented in histograms it is much easier to see.

It may be that both signals cannot be collected at a similar intensity as to observe a nice mix of both (like a nice yellow from red and green mixing). For profile scan it doesn't matter because you just want to show that the position of each peak is similar, whatever the height of the peaks. But for mixed images it is of course better if the structure has a well balanced mix of both signals (say the red is much stronger than the green, the merged image shows a red structure; well it is not a very convincing demonstration of colocalization).
Thus it is often required to increase the intensity of one signal. Take care to increase the overall intensity of the whole picture and not only part of the histogram (like playing with the contrast) or even worse: only a part of the image (this is a misconduct). If playing with the contrast is acceptable, do not forget to state it but I would avoid it.

Good luck
 
Stephane

----- Original Message ----
X-from: "kjmorris-at-well.ox.ac.uk" {kjmorris-at-well.ox.ac.uk}
To: nizets2-at-yahoo.com
Sent: Thu, June 10, 2010 11:07:29 AM

Hi Molly

Just to add to Stephane's and Jerry's comments. You are right that a
confocal will not benefit from deconvolution software anywhere near as much
as a widefield microscope, for the reason you suggest, i.e. much of the
out-of focus information is removed by the confocal iris/pinhole in any
case. I use Improvison's [Perkin Elmer] 3D Volocity software to look at
things like 3D images, 3D cellular volumes and cell morphometry. In order to
get the best from a Z slice you are advised to over sample the z-stack, i.e.
for a 0.9um optical slice capture the z stack at least 0.45um apart [our
Zeiss 510 has a button to automatically 'double oversample' like this]. For
the likes of Volocity deconvolution, Improvision recommended to me that you
massively oversample the confocal z-stack with ten or more slices to every
0.9um z focus step [I think the rep suggested perhaps up to as high as 100x
oversampling]. I rarely use the optional 'Restoration' module
[deconvolution] Volocity module as I only use the confocal for z-stack, and
double oversampling and 3D reconstruction alone with Volocity is adequate
for my needs.

For co-localisation I would agree with Stephane though, and initially try
and tackle the co-localisation quantification in 2D using MetaMorph
[probably just because it's just easier]. I don't use ImageJ very much as I
have a MetaMorph licence, and we don't have the 3D Quantitation module for
Volocity in any case, that might be needed for 3D co-localisation
quantification - if interested just ask Improvision about that as their
support is superb.

Always be suspicious that any co-localisation isn't bleedthrough. You can
check this via spectral un-mixing on a 510 Metahead or just try imaging
single labeled samples of each fluorochrome and check there's no signal in
the other channels where there is now no longer any label [at the same
confocal imaging settings]. Don’t' forget a sample with no label just in
case of autofluorescence.

To quote Improvisions own website: "If you acquire your images from a
widefield microscope and want to volume render or make measurements from the
image data, you will need to use an image restoration technique
[de-convolution] to remove the out-of-focus information - a product of the
optical properties of the microscope. Even if your images are captured using
a laser scanning or spinning disk confocal microscope, the image quality may
benefit from image restoration." The word 'may' is probably significant.

Regards

Keith
---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford  OX3 7BN,
United Kingdom.

Telephone:  +44 (0)1865 287568
Email:  kjmorris-at-well.ox.ac.uk
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy

-----Original Message-----
X-from: mshaw1-at-lumc.edu [mailto:mshaw1-at-lumc.edu]
Sent: 07 June 2010 14:37
To: kjmorris-at-well.ox.ac.uk

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Email: mshaw1-at-lumc.edu
Name: Molly Shaw

Organization: Loyola University Chicago

Title-Subject: [Filtered] deconvolution & confocal microscopy

Message: Hello,

I am a first-year graduate student trying to get started with
colocalization analysis of IF images taken by confocal microscopy. I
am working with fixed cells and mainly trying to measure
colocalization of 2 proteins. My main questions are concerning the
use of z-stacks and deconvolution:
1. Are z-stacks always necessary for measurement of colocalization,
or are single images sufficient?
2. I understand that deconvolution is essential for wide-field
images, but does it improve results that much when you're already
using confocal?
3. If you would recommend deconvolution, is the ImageJ plug-in a good
one to use?

Thank you for your time & help. I have read many articles on this
topic, but I keep ending up with conflicting answers.

Thanks again,
Molly Shaw

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From: thoward-at-unm.edu
Date: Fri, 11 Jun 2010 12:43:03 -0500
Subject: [Microscopy] Definiens Tissue Studio?

Contents Retrieved from Microscopy Listserver Archives
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Apologies to everyone on the cross-post for this question,
but I'm looking for feedback on this image analysis
package (Definiens Tissue Studio). One of our faculty
members is interested in it & I know nothing about it,
other than what the company website has to offer; I'd
rather hear from people who actually use it! Their group
looks at a variety of sample types - whole mounts,
cultured cells, & sections by both fluorescence (epi &
confocal) and brightfield microscopy. Pros, cons, any
insights will be appreciated. Feel free to reply off-line;
I won't post any replies I receive in confidence.

Thanks!

Tamara

***************************
Tamara Howard
Cell Biology & Physiology
UNM-HSC
Albuquerque, NM
***************************


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From: sarj0007-at-unf.edu
Date: Fri, 11 Jun 2010 15:06:27 -0500
Subject: [Microscopy] Ultra microtome manual

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

Thank you everyone who has already responded. We tracked down sections 2-5 so should be all set now. Have a great weekend.

-Jason

________________________________________
X-from: sarj0007-at-unf.edu [sarj0007-at-unf.edu]
Sent: Thursday, June 10, 2010 12:18 PM
To: Saredy, Jason

I was wondering if anyone had an electronic copy of the manual for the Serval Porer-Blum MT-2, or know where to track one down. We have dragged ours out of the closet and want to double-check some of the knobs are working like they should be. Thanks.

-Jason Saredy


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From: gtuser-at-comcast.net
Date: Sat, 12 Jun 2010 18:34:20 -0500
Subject: [Microscopy] viaWWW: Quantify the Backscatter Electron intensities

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Email: gtuser-at-comcast.net
Name: Tommy Derflinger

Organization: URS at NSWC Crane

Title-Subject: [Filtered] Quantify the Backscatter Electron intensities

Message: From a BSE image can the yield or intensities be quantified
to give a fairly accurate number?

It will be on a Bianary alloy of metals.

Does anyone know if that can be done?

Thank you

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From: gary-at-gaugler.com
Date: Sat, 12 Jun 2010 19:57:30 -0500
Subject: [Microscopy] Re: viaWWW: Quantify the Backscatter Electron

Contents Retrieved from Microscopy Listserver Archives
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I would think so but only for that particular capture.
Which would mean that it is actually relative to the
other elements in the image. One would need a known
standard along with the specimen to make any headway
towards a meaningful quantification. In any case,
quantification would just mean what the pixel intensity
is at any point on the image. I don't see what use this
would be other than to say that element x is twice the
intensity of element y. Interesting, but so what?

Just as with film and a densitometer, contrast and brightness
will change all elements' intensity. Of course one can obtain
the same type of "quantitative" results but again, they are
relative to that piece of film and/or print.

What is the end purpose? I would think that EDS mapping
is going to provide true quantitative results for a specimen.
Having counts provided at each pixel is quantitative.

gary g.


At 04:36 PM 6/12/2010, you wrote:



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From: kqolizadeh-at-yahoo.com
Date: Sun, 13 Jun 2010 09:09:08 -0500
Subject: [Microscopy] viaWWW: EPMA problems

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Email: kqolizadeh-at-yahoo.com
Name: kazem

Organization: IMPRC

Title-Subject: [Filtered] EPMA problems

Message: Hi dear friends
We've bought a SX 100 EPMA(tungsten filament) at a gov't auction and
are trying to get it up and running. We dont have any devices like
EPMA and now it doesnt work and all electronics are off. Nobody knows
it exactly in our country but most electronicion guess cause of
chiller.(does it have cooling that might be plugged or not?).
We have it pumping, can get filament current and think we have the
detector's configured properly, but there is no beam/signal.

Does someone have the same tool that might be able to answer some
basic questions? We're not even sure if we have all the parts (it was
missing a pump and other stuff?) and some quick pictures of the
plumbing would be very helpful. Same thing with the 'low KV anode'-
if we open up the gun how do we recognize what we have?

Does anyone have a source for technical documents for this tool? The
online help leaves much to be desired.

thanks,


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From: hstahlberg-at-me.com
Date: Mon, 14 Jun 2010 05:30:00 -0500
Subject: [Microscopy] Reminder: Workshop on Electron Crystallography of Membrane Proteins:

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


Hi,
A reminder for the upcoming third Workshop on Electron Crystallography
of Membrane Proteins, August 1-7, 2010, in C-CINA, University Basel,
Switzerland.
Registration is open until June 15, 2010.

More info and Registration is at
http://www.2dx.unibas.ch/workshop/2010

all the best,

Tom, Andreas, Karen, and Henning.


___________________________________________________

Henning Stahlberg
Center for Cellular Imaging and Nano Analytics (C-CINA)
at the Department of Biosystems Science and Engineering (D-BSSE)
Structural Biology and Biophysics, Biozentrum,
WRO-1058, Mattenstrasse 26
University Basel, CH-4058 Basel, Switzerland
Tel: +41-61-387 32 62 (office)
Tel: +41-61-387 32 31 (Karen Bergmann, administrative assistant)
Fax: +41-61-387 39 86
mailto:Henning.Stahlberg-at-unibas.ch
Skype:henningstahlberg
http://c-cina.org
http://2dx.org
___________________________________________________

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From: DusevichV-at-umkc.edu
Date: Mon, 14 Jun 2010 09:38:27 -0500
Subject: [Microscopy] RE: viaWWW: Quantify the Backscatter Electron intensities

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

You may want to check this paper:

Validation of quantitative backscattered electron imaging for the
measurement of mineral density distribution in human bone biopsies
P. Roschger, P. Fratzl, J. Eschberger and K. Klaushofer,
Bone, Volume 23, Issue 4, October 1998, Pages 319-326

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784


} -----Original Message-----
} From: gtuser-at-comcast.net [mailto:gtuser-at-comcast.net]
} Sent: Saturday, June 12, 2010 6:36 PM
} To: Dusevich, Vladimir
} Subject: [Microscopy] viaWWW: Quantify the Backscatter Electron
} intensities
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} Title-Subject: [Filtered] Quantify the Backscatter Electron
intensities
}
} Message: From a BSE image can the yield or intensities be quantified
} to give a fairly accurate number?
}
} It will be on a Bianary alloy of metals.
}
} Does anyone know if that can be done?
}
} Thank you
}
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intensities
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From: jfmjfm-at-umich.edu
Date: Mon, 14 Jun 2010 09:49:48 -0500
Subject: [Microscopy] Queen's Birthday Honours List 2010

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

I have not seen anyone note this yet, but two microscopists were honoured in the UK on the Queen's Birthday Honours List.
For a full list see the Daily Telegraph page here:

http://www.telegraph.co.uk/news/newstopics/honours-list/7822829/Queens-Birthday-honours-the-full-list.html

But the two of note to microscopy folks are:

Order of the British Empire
DBE Professor Athene Margaret Donald, FRS. Deputy head, Cavendish Laboratory and director, Women in Science, Engineering and Technology Initiative, University of Cambridge. For services to Physics. (Cambridgeshire)

and

Knights Bachelor

Professor Colin John Humphreys, CBE. Director of Research, Department of Materials Science and Metallurgy, University of Cambridge. For services to Science.

Congratulations to them both.

--
John Mansfield PhD Cphys MInstP
2010 Microscopy & Microanalysis Program Chair
North Campus Electron Microbeam Analysis Laboratory
417 SRB, University of Michigan
2455 Hayward, Ann Arbor MI 48109-2143 USA
Phone: (734) 936-3352 FAX (734) 763-2282
Cell. Phone: (734) 834-3913
(Leaving a phone message at 936-3352 is preferable to 834-3913)
Email: jfmjfm-at-engin.umich.edu
URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
Location: N 42 ° 17' 38.3" (42 ° 17.6391' ) W 83 ° 42' 38.6" (-83 ° 42.6439')
AIM: thejfmjfm
Yahoo: thejfmjfm
Skype: thejfmjfm

Home address:
4304 Spring Lake Boulevard
Ann Arbor MI 48108-9657
Phone (734) 994-3096
Location: N 42 ° 13' 28.8" (42 ° 13.4807') W 83 ° 45' 47.9" 42° 16' 48" (-83 ° 45.7980')

Please note: Electronic Mail is not secure, but should be read several times every day, and should definitely be used for urgent or sensitive issues.



==============================Original Headers==============================
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From: b-myers3-at-northwestern.edu
Date: Mon, 14 Jun 2010 10:50:13 -0500
Subject: [Microscopy] Re: viaWWW: Quantify the Backscatter Electron intensities

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

I designed a lab exercise for one of our courses here that uses BSE
contrast to quantitatively determine the thickness of a metal film on a
substrate. Keep in mind that this is intended as an educational
exercise, but I've been surprised by the accuracy of the results.

The idea in the lab is to determine the thickness of a Au/Pd film
(patterned by eBL on silicon) by measuring the relative contrast from
the Si substrate, Au/Pd film on substrate and bulk Au/Pd. The students
can determine the effective BSE coefficient for the film-on-substrate by
using the calculated coefficient from the bulk materials and the
relative contrast values. Then by using a Monte Carlo simulation for a
film-on-substrate, they can back out the film thickness by varying the
thickness in the simulation and matching the BSE coefficient to the
experimentally determined value. Using this method we have been able to
determine the thickness of this ~25nm film within 15 %...

Here are some pitfalls:

1) You need to reference a standard(s) with the SAME imaging conditions
(contrast/brightness, WD, kV, beam current, etc.)
2) Any surface roughness will alter the average contrast values due to
BSE variation with incident angle.
3) You need a surface free of any contamination.
4) The dynamic range in your contrast measurements will strongly impact
the error in your measurements. If you can capture 16-bit images, this
would help.
5) One thing I've never been sure about is the linearity of contrast vs.
BSE intensity for different detectors. However, it should be relatively
easy to calibrate..

Hope that helps to some extent.

Regards,
Ben


On 6/12/2010 6:46 PM, gtuser-at-comcast.net wrote:
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} Email: gtuser-at-comcast.net
} Name: Tommy Derflinger
}
} Organization: URS at NSWC Crane
}
} Title-Subject: [Filtered] Quantify the Backscatter Electron intensities
}
} Message: From a BSE image can the yield or intensities be quantified
} to give a fairly accurate number?
}
} It will be on a Bianary alloy of metals.
}
} Does anyone know if that can be done?
}
} Thank you
}
} Login Host: 98.223.31.103
} ---------------------------------------------------------------------------
}
} ==============================Original Headers==============================
} 9, 11 -- From zaluzec-at-microscopy.com Sat Jun 12 18:34:20 2010
} 9, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22])
} 9, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id o5CNYId6021819
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} 9, 11 -- Date: Sat, 12 Jun 2010 18:34:18 -0500
} 9, 11 -- To: microscopy-at-microscopy.com
} 9, 11 -- From: gtuser-at-comcast.net (by way of MicroscopyListserver)
} 9, 11 -- Subject: viaWWW: Quantify the Backscatter Electron intensities
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}

--
*Ben Myers*
SEM/FIB Facility Manager
NU/ANCE/ Center

Northwestern University
Mail: 2036 Cook Hall
Office: 1114 Cook Hall
2220 Campus Drive
Evanston, IL 60208-3108

ph: (847) 491-3439
fax: (847) 467-6573

http://www.nuance.northwestern.edu

==============================Original Headers==============================
12, 18 -- From b-myers3-at-northwestern.edu Mon Jun 14 10:50:13 2010
12, 18 -- Received: from mailp7.ci.northwestern.edu (casbah.it.northwestern.edu [129.105.16.97])
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From: kenconverse-at-qualityimages.biz
Date: Mon, 14 Jun 2010 15:02:39 -0500
Subject: [Microscopy] Re: viaWWW: Quantify the Backscatter Electron

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

It seems to me that between 20 and 30 years ago there was a LOT of work done
on quantitative backscatter with many results showing discrimination of
compounds down to about 0.1 atomic mass units. As I recall it was quite
reproducible and quantitative.

My guess is that it was not pursued more due to the performance improvements
and cost reductions in EDS.

I'd love to see comments posted from those who have done some of this.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: b-myers3-at-northwestern.edu [mailto:b-myers3-at-northwestern.edu]
Sent: Monday, June 14, 2010 11:53 AM
To: kenconverse-at-qualityimages.biz

I designed a lab exercise for one of our courses here that uses BSE
contrast to quantitatively determine the thickness of a metal film on a
substrate. Keep in mind that this is intended as an educational
exercise, but I've been surprised by the accuracy of the results.

The idea in the lab is to determine the thickness of a Au/Pd film
(patterned by eBL on silicon) by measuring the relative contrast from
the Si substrate, Au/Pd film on substrate and bulk Au/Pd. The students
can determine the effective BSE coefficient for the film-on-substrate by
using the calculated coefficient from the bulk materials and the
relative contrast values. Then by using a Monte Carlo simulation for a
film-on-substrate, they can back out the film thickness by varying the
thickness in the simulation and matching the BSE coefficient to the
experimentally determined value. Using this method we have been able to
determine the thickness of this ~25nm film within 15 %...

Here are some pitfalls:

1) You need to reference a standard(s) with the SAME imaging conditions
(contrast/brightness, WD, kV, beam current, etc.)
2) Any surface roughness will alter the average contrast values due to
BSE variation with incident angle.
3) You need a surface free of any contamination.
4) The dynamic range in your contrast measurements will strongly impact
the error in your measurements. If you can capture 16-bit images, this
would help.
5) One thing I've never been sure about is the linearity of contrast vs.
BSE intensity for different detectors. However, it should be relatively
easy to calibrate..

Hope that helps to some extent.

Regards,
Ben


On 6/12/2010 6:46 PM, gtuser-at-comcast.net wrote:
}
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Listserver
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---------------------------------------------------------------------------
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} Email: gtuser-at-comcast.net
} Name: Tommy Derflinger
}
} Organization: URS at NSWC Crane
}
} Title-Subject: [Filtered] Quantify the Backscatter Electron intensities
}
} Message: From a BSE image can the yield or intensities be quantified
} to give a fairly accurate number?
}
} It will be on a Bianary alloy of metals.
}
} Does anyone know if that can be done?
}
} Thank you
}
} Login Host: 98.223.31.103
}
---------------------------------------------------------------------------
}
} ==============================Original
Headers==============================
} 9, 11 -- From zaluzec-at-microscopy.com Sat Jun 12 18:34:20 2010
} 9, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com
[206.69.208.22])
} 9, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id
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} 9, 11 -- To: microscopy-at-microscopy.com
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} 9, 11 -- Subject: viaWWW: Quantify the Backscatter Electron intensities
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}

--
*Ben Myers*
SEM/FIB Facility Manager
NU/ANCE/ Center

Northwestern University
Mail: 2036 Cook Hall
Office: 1114 Cook Hall
2220 Campus Drive
Evanston, IL 60208-3108

ph: (847) 491-3439
fax: (847) 467-6573

http://www.nuance.northwestern.edu

==============================Original Headers==============================
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(casbah.it.northwestern.edu [129.105.16.97])
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From: sedge001-at-umn.edu
Date: Tue, 15 Jun 2010 09:24:04 -0500
Subject: [Microscopy] 1st Annual Imaging in Research Course (commercial-academic announcement)

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

Here's why the 1st Annual Imaging in Research Course is being given in
August, 2010:

* Visual data in the form of images is often 50 percent of a manuscript's
content. It's content is a reflection of the lab, and so the quality and
accuracy of images is as important as the written content.

* 44 percent of cases investigated by the Office of Research Integrity in
2005-6 involved accusations of image fraud, compared with about 6 percent a
decade before that (1). These cases are rising, and most involve graduate
and post-doctoral students.

* Students and staff who self-report familiarity with imaging programs like
Photoshop may be using it improperly.

In response to this, the "1st Annual Imaging in Research Course: Ethics,
Acquisition, Post-Processing, Output and Segmenting" is being held at the
University of Minnesota Continuing Education Center in Minneapolis/St.
Paul, Minnesota, August 16 - 19, 2010, sponsored by the Histochemical
Society and the Adobe Corporation. Attendees can choose to attend the
course for 1- to 4-days. This workshop will educate those in science,
medicine and engineering about correct techniques when acquiring,
post-processing, and adjusting images for outputs; along with techniques
that work for segmenting complex, biological images (for subsequent image
analysis).

Other benefits of taking the course will likely result in:

Faster acceptance of submitted manuscripts
Authors better able to demonstrate outcomes to their target audience
Faster results from quantitation, with improved ability to segment desired
features
Better documentation of imaging procedures
Standardization of post-processing
Learning to adjust and modify images minimally and through the objective
use of numbers.

Jerry Sedgewick will present, along with invited speakers. Jerry directed a
core light microscopy and imaging facility for 15 years at the University
of Minnesota, published 2 books on Photoshop and digital imaging, and his
quantitative work has led to FDA approval for start up companies.

Please go to http://www.imagingandanalysis.com/seminars.html for more
information. There is a limit of 30 seats.

The cost ranges from $195 for 1-day to $840 for 4-days. It includes lunch,
beverages and snacks. Registrations for those who received information
about this course via their core facility will receive discounts. Here's
the summary for the days:

Day1: Ethics of digital imaging, sample preparation, calibration, best
acquisition practices on light microscopes.
Day2: Post Processing I: setting up Photoshop, opening image stacks/12 bit
images, rotate/crop, uneven illumination correction, color correction,
histogram (tone) matching, gamma corrections, correcting noise, scale bars,
extended focus, extended dynamic range, pseudocolor, tonal adjustment.
These functions also covered for the free programs GIMP and Image J (when
applicable).
Day 3: Post Processing II: De-colorizing/colorizing fluorescent samples,
merging images, colocalization, adjusting tones for 3D reconstructions,
saving images, resetting pixel resolution (resampling), creating automated
steps (macros), image stitching, making figures, better methods for
sharpening, digital video, images to various outputs. These functions also
covered for the free programs GIMP and Image J (when applicable).
Day 4: Segmenting in Photoshop for image analysis (quantitation): optical
density and intensity measurements, creating binary files through 3
methods, setting threshold at consistent value, unbiased sampling
(stereology), automating steps, measurement in Image J and Excel.

1. "Journals Find Many Images in Research Are Faked," Jeffrey R. Young, The
Chronicle of Higher Education, September 9, 2009)

All the best,

Jerry Sedgewick

==============================Original Headers==============================
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From: maloneyb-at-fiu.edu
Date: Wed, 16 Jun 2010 13:39:57 -0500
Subject: [Microscopy] Cambridge Galen III - light microscope

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

Dear Group - would anyone by chance have the manual for this? Would
anyone know the name of the tool that tightens the flush knob for the
condenser?
Thanks so much.
Barbara

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From: kenconverse-at-qualityimages.biz
Date: Fri, 18 Jun 2010 16:29:46 -0500
Subject: [Microscopy] SEM

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

The Facilities Operation and Management Focus Interest Group of MSA shall
have its Annual Luncheon, Business Meeting and Round Table Discussion this
year during M&M 2010 on August 3 in Room A108 Oregon Convention Center,
Exhibit Level.

All in attendance at M&M 2010 are welcome to participate in the Round Table
Discussion starting at 12:30. Owen Mills, who was elected Leader of the
FOM FIG for 2010 - 2011, will lead these discussions.

The topics for the 2010 Roundtable Discussion are:
1. What happens when a user breaks something that is not covered by service
contracts?
2. How do we best educate administrators as to the needs of our facilities
3.Conflict Management

If you are wondering if you are a current member of the FOM FIG you ARE IF
you already received a similar message that included information on the
Luncheon.
If you did not receive that message please join MSA and sign up for our FIG.
If you have paid your MSA dues for this year, you can join the FOM FIG by
sending in the $10.00 additional dues to MSA and by sending an email message
to {psconnelly-at-gmail.com} so I will know to ask for an updated membership
list and be able to send you the additional information on the luncheon and
business meeting.

Early Registration for M&M 2010 ends on JUNE 21st!

Pat Connelly

FOM FIG Secretary
{psconnelly-at-gmail.com}
(please use this email address and do not reply to my posting address)



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From kefwysoccydiz-at-wysoccy.com Fri Jun 18 05:39:44 2010
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We have a s-510 Hitachi SEM. We have no high voltage from our power
supply to the crt. We have tested all the voltages on the board for the
high voltage and everything tests correctly. The high voltage supply is
oil filled and we were trying to avoid opening it up. Has any one
encountered this and if so how did you resolve.


Aaron Davison
Manufacturing Support Technician
Resonant Microsystems
2322 Alpine Rd
Mailbox #4
Eau Claire, Wi 54703
715-874-6800
adavison-at-resonantmicro.com




==============================Original Headers==============================
6, 19 -- From adavison-at-resonantmicro.com Fri Jun 18 08:30:11 2010
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From kefwynonnadiz-at-wynonna.com Fri Jun 18 09:21:53 2010
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Aaron,
I'm not terribly familiar with the S510, and some SEMs do put all their HV
supplies in the oil tank, but many (most?) don't because the CRTs require a
positive 10-12 kV. The gun requires a negative 25-40 (sometimes 50)kV so
now you're having to isolate up to 60kV in one unit.

Have you traced the CRT HV lead to the oil tank? If you have, then you may
have to go into the tank. It shouldn't be too bad. First check and make
sure that the S510 is post-PCB ban, which it probably is. After that, get
lots of paper towels (bench pads are terrific if you have them) and just
move slowly so most of the oil drains right back into the tank. You should
have pretty good access to the high voltage side of things.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: adavison-at-resonantmicro.com [mailto:adavison-at-resonantmicro.com]
Sent: Friday, June 18, 2010 9:38 AM
To: kenconverse-at-qualityimages.biz

We have a s-510 Hitachi SEM. We have no high voltage from our power
supply to the crt. We have tested all the voltages on the board for the
high voltage and everything tests correctly. The high voltage supply is
oil filled and we were trying to avoid opening it up. Has any one
encountered this and if so how did you resolve.


Aaron Davison
Manufacturing Support Technician
Resonant Microsystems
2322 Alpine Rd
Mailbox #4
Eau Claire, Wi 54703
715-874-6800
adavison-at-resonantmicro.com




==============================Original Headers==============================
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==============================Original Headers==============================
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From: kwilliam-at-mun.ca
Date: Sat, 19 Jun 2010 01:37:39 -0500
Subject: [Microscopy] viaWWW: Digital cmaera for JEOL 1200EX and quality control in EM

Contents Retrieved from Microscopy Listserver Archives
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Email: kwilliam-at-mun.ca
Name: Kate Williams

Organization: Memorial University

Title-Subject: [Filtered] Digital cmaera for JEOL 1200EX and quality
control in EM Lab

Message: I operate a JEOL 1200EX, we are interested in purchacing a
digital camera for this instrument. I have narowed my wish list down
to two criterion side mounted and at least 4 megapixel.I have the
manufactures literature but am looking for comments and suggestions
based on personal practical experience. Does anyone have suggestions
that would help me make a final wish list . We do clinical and
biological research.
On another completely unrelated topic does anyone know of a quality
assurance program for clinical electron microscopy labs. I can not
find anything on line but am interested in what other labs may do as
internal controls.
Thank you
Kate

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From: dsherman-at-purdue.edu
Date: Sun, 20 Jun 2010 21:34:02 -0500
Subject: [Microscopy] Yeast slides for LM

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Email: jacqueline.ayotte-at-ticona.com
Name: Jackie Ayotte

Organization: Ticona Engineering Polymers

Title-Subject: [Filtered] SEM of air sensitive samples

Message: Hello All,

I am challenged with the task of performing SEM on air sensitive
catalysts. Can anyone share their method for doing this? I have seen
SEM images of the catalysts that I am interested in analyzing.
However the company that generated those micrographs can not share
their procedure with me.

I can provide more information regarding the catalyst off-line.

Much Thanks -

Jackie

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From jadams-at-nls.k12.la.us Sun Jun 20 05:14:12 2010
Return-Path: {jadams-at-nls.k12.la.us}
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Hi all,

Our beginning Biology course has a lab that requires slides for counting
cells using a micrometer eyepiece on a light microscope. They want to use
yeast but found that preparing fresh samples during the lab took too much
time and was so variable that the students did not get to complete the lab
on time. Thus they would like to make permanent slides that can be used in
the labs for a number of years.

Suggestions as to how to prepare slides (about 50 of them) containing a
monolayer of yeast stained with hemotoxylin would be appreciated.

Debby


---
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy




==============================Original Headers==============================
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From: naomi_mccallum-at-health.qld.gov.au
Date: Sun, 20 Jun 2010 22:47:51 -0500
Subject: [Microscopy] Fwd: Yeast slides for LM

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

Hi Debby

I realise that you are probably referring to growing the monolayer on the slides but here's a response from left field. Perhaps a cytology lab could assist with preparing Cytospin or ThinPrep slides (depending on the no. of slides required). You may have to adjust the yeast solution cell count slightly to achieve the desired monolayer. And perhaps the standard PAP stain may suit your work but I think they usually use Periodic Acid Schiff stain in diagnosis.

Naomi

} } } {dsherman-at-purdue.edu} 21/06/2010 12:44 pm } } }



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Hi all,

Our beginning Biology course has a lab that requires slides for counting
cells using a micrometer eyepiece on a light microscope. They want to use
yeast but found that preparing fresh samples during the lab took too much
time and was so variable that the students did not get to complete the lab
on time. Thus they would like to make permanent slides that can be used in
the labs for a number of years.

Suggestions as to how to prepare slides (about 50 of them) containing a
monolayer of yeast stained with hemotoxylin would be appreciated.

Debby


---
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy




==============================Original Headers==============================
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9, 28 -- Date: Sun, 20 Jun 2010 22:33:59 -0400
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==============================Original Headers==============================
20, 26 -- From prvs=17887cd7f6=naomi_mccallum-at-health.qld.gov.au Sun Jun 20 22:47:51 2010
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20, 26 -- To: {Microscopy-at-microscopy.com}
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From: kamlennon-at-yahoo.com
Date: Mon, 21 Jun 2010 11:01:33 -0500
Subject: [Microscopy] SEM-Materials Demo

Contents Retrieved from Microscopy Listserver Archives
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Dear Debby,

If you just want to count small things, you could use pollen. I think you
can just mount it in nailpolish, coverslip, and let it dry. These last
quite a while. Any abundantly productive plant will do. Mushroom spores
are pretty abundant and hardy as well and also small - reasonably fresh
mushroom cap, gills down, on white paper overnight or a bit longer will
provide plenty of spores.

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601

T 02 6246 5475
F 02 6246 5334
E rosemary.white-at-csiro.au


On 21/06/10 1:53 PM, "naomi_mccallum-at-health.qld.gov.au"
{naomi_mccallum-at-health.qld.gov.au} wrote:

}
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}
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} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi Debby
}
} I realise that you are probably referring to growing the monolayer on the
} slides but here's a response from left field. Perhaps a cytology lab could
} assist with preparing Cytospin or ThinPrep slides (depending on the no. of
} slides required). You may have to adjust the yeast solution cell count
} slightly to achieve the desired monolayer. And perhaps the standard PAP stain
} may suit your work but I think they usually use Periodic Acid Schiff stain in
} diagnosis.
}
} Naomi
}
} } } } {dsherman-at-purdue.edu} 21/06/2010 12:44 pm } } }
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
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} Hi all,
}
} Our beginning Biology course has a lab that requires slides for counting
} cells using a micrometer eyepiece on a light microscope. They want to use
} yeast but found that preparing fresh samples during the lab took too much
} time and was so variable that the students did not get to complete the lab
} on time. Thus they would like to make permanent slides that can be used in
} the labs for a number of years.
}
} Suggestions as to how to prepare slides (about 50 of them) containing a
} monolayer of yeast stained with hemotoxylin would be appreciated.
}
} Debby
}
}
} ---
} Debby Sherman, Director Phone: 765-494-6666
} Life Science Microscopy Facility FAX: 765-494-5896
} Purdue University E-mail: dsherman-at-purdue.edu
} S-052 Whistler Building
} 170 S. University Street
} West Lafayette, IN 47907
} http://www.ag.purdue.edu/facilities/microscopy
}
}
}
}
} ==============================Original Headers==============================
} 9, 28 -- From dsherman-at-purdue.edu Sun Jun 20 21:34:02 2010
} 9, 28 -- Received: from mailhub130.itcs.purdue.edu (mailhub130.itcs.purdue.edu
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} 9, 28 -- To: "message to: MSA list" {microscopy-at-microscopy.com}
} 9, 28 -- Date: Sun, 20 Jun 2010 22:33:59 -0400
} 9, 28 -- Subject: Yeast slides for LM
} 9, 28 -- Thread-Topic: Yeast slides for LM
} 9, 28 -- Thread-Index: AcsQ6jPwtA3LKK+gJ06LJ5sNOAyjmg==
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} including any attachment sent with it, may be subject to a statutory duty of
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} 20, 26 -- From prvs=17887cd7f6=naomi_mccallum-at-health.qld.gov.au Sun Jun 20
} 22:47:51 2010
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} 20, 26 -- From: "Naomi McCallum" {naomi_mccallum-at-health.qld.gov.au}
} 20, 26 -- To: {Microscopy-at-microscopy.com}
} 20, 26 -- Subject: Fwd: [Microscopy] Yeast slides for LM
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From bridgette_carey-at-mail.cider.com Mon Jun 21 03:32:04 2010
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Hi Stephane and Molly,

Sorry I didn't intend to comment on 'quantifying' colocalisation at all in my last post, just on the confocal and deconvolution 'image sharpening' aspects of the question [Stephane covered the colocalisation aspects in any case]. I may as well add my thoughts though on colocalisation [also see the links at the end, e.g. the article "Quantitative analysis of fluorescently labeled nuclear structures: Problems, methods, pitfalls" - and I assume the 'IF' in your post stands for immunofluorescence]:

We have various 'colocalisation' 2D image analysis software packages here that still probably more 'suggest' rather than accurately 'quantify' colocalisation, despite the colocalisation 'results' being given as an exact figure to one decimal place [see 'intensity measurements' below]. Our MetaMorph software for example offers "Colocalization tools to provide intensity measurements of the region of overlap between two fluorescent probes", although there's many other image analysis packages [I've not used] as well, such as Imaris [ImarisColoc], IMAJIN_COLOC, Image Pro Plus Colocalization module, ImageJ's colocalization plug-ins, and often the confocal itself has specific tools for visualizing 'colocalisation' as well [ask the reps]. Also try Stephane's suggestions, and with some software you can draw your intensity profile lines in 3D through the z-stack.

You want to be quite precise with your z-plane optical slice [Airy=1] to ensure that the regions of fluorochrome overlap are genuinely in pretty much the same place [so a 63x Zeiss Plan Apochromat objective would definitely be preferred to a 40x Zeiss Plan Neofluar objective]. I personally don't see a problem with analyzing in 2D, i.e. one z plane only, as you are just sampling a small subset of the cell population so why not a subset of the cell as well - if your cells are plated down flat you could measure 2D colocalisation in a few of the z planes as there probably won't be so many of them. Or if you're looking for a new hobby you could analyse all the z slices [MetaMorph does the analysis pretty quickly], in effect giving you the 3D volume co-localisation as you know the XY pixel size and approx z depth slice thickness in um]. Number of cells to look at, well 30 should be adequate, but the more the better for the stats. You can also use the likes of Volocity's quantitation module to analyse co-localisation in the 3D volume or 4D [volume+time] directly, and other 3D packages probably offer similar. ImageJ has freebie colocalisation plug-ins, sorry I don't use ImageJ but check out the ImageJ link below [and it's likely a licensed copy of MetaMorph or Image Pro Plus will be about on any biomedical research campus as well]. There's Image J's Colocalization, RG2B Colocalization, and Colocalization_Finder plugins available.

Just seeing a yellow region in the red/green overlap area image isn't really telling you a lot about colocalisation [and it's not quantifying it, i.e. how much of the total red and green intensity signal throughout the image is actually located in these areas of overlap and how much isn't]. Colocalisation measurements try to quantify both the fluorochrome emission intensities as well as their location in the sample, and colocalisation applications work through the image or z-stack pixel by pixel, giving the results not just for the regions of overlap but also the regions where only one flourochrome or the other is present at a significant concentration [as determined by emission signal intensity]. Often the results are simply presented as percentage overlap, i.e. how much of the total A [green] + B [red] fluorescent intensity is or isn't located within the same pixels. For example MetaMorph's colocalisation app reports colocalisation as A overlapping B, A not overlapping B, B overlapping A, and B not overlapping A [all values as either percentage or um2 area]. It also gives the pixel intensity measurements as either mean pixel intensity [i.e. 'concentration'] or 'integrated: the sum of all pixel intensities in the bright regions' [i.e. 'mass'], and it reports the total areas of A and B in um2. User input is often required for selecting [thresholding] the bright regions and capturing them in the first place, so 'quantitative' colocalisation results from the same sample may well vary between users, but hopefully all would show a similar trend. Some microscope and image analysis software presents these measurements of regions of overlap as 'colocalisation coefficients'.

Search on-line for more info on colocalisation [e.g. see links below]. Back in 2004 I posted comments on the listserver about quantifying intra-cellular fluorescence intensity to compare brightness between different samples. It's not about colocalisation measurement as such but the problems discussed are still relevant, although Colocalisation adds in the additional complication of collecting two or perhaps three quite different excitation/emission channels into each pixel so you have to plan your sample preparation, image acquisition and sampling/analysis regime carefully and consistently:

Intensity measurements

"It's not really possible to get quantitative measurements of epi-fluorescence. The gel fluorescence standards commonly suggested are often used to check that your camera/PMT detector and electronics are genuinely linear in response (something quite important to know) - they are no use as standards for calibrating fluorescence within the cell (although you can use things like BCECF in solution at known pH's to roughly calibrate intracellular pH).

To properly calibrate cellular fluorescence you would need standards that are essentially known concentrations and masses of labelled protein etc.. not something that's easy to get. When comparing intensities between samples you will always have problems with confocal laser power variation, detector linearity, confocal electronic stability, biological variation, differential bleaching, uneven inconsistent labelling or background, non-specific labelling, optical effects, internal quenching, etc.. so you will probably never be able to say that one cell has twice the mass (total pixel brightness) or concentration (mean pixel brightness) of a given labelled protein or whatever compared to another. However it is reasonable to assume, given identical preparation and imaging procedures, that a very brightly labelled cell has definitely more labelled fluorochrome in it than a weakly fluorescent cell or intracellular region (and you could say something like 'suggesting' x times concentration etc.). Also try measuring something simple like concentrations of fluorescent beads to see how (badly) the image analysis results work out (these do have the advantage that you can actually count them as well if they are big enough).

You will probably have to use regions of interest to include darker as well as bright areas if comparing intensity between samples, plus you can use peak values as described by Stephane (but often the bright sample fluorescence area has to be clearly defined to one region to get this to work [note this refers to a 2004 post]). You can also try things like what % of the cell is brighter than a set grey level. Co-localisation (i.e. the % of bright pixels in both the red and green channel across the cell) is also useful, MetaMorph has a simple app to calculate this. Generally each set of samples require their own image analysis protocols depending on what you want to find out, plus you need simple stats to say if the difference is significant to 95% level. As always consistent specimen preparation quality is particularly important.

When looking at optical density, this can in theory be more easily calibrated - here you are measuring optical white light transmission though a sample. So you can create black (metal disk), white (free space) and have a selection of known density materials like polythene (grey) etc. standards within the image. You can therefore assign a grey level to a particular density. We have used this to estimate things like bone density, assuming you can get very evenly sliced samples [i.e. using a small annular histological bone saw]."

You would have to be careful about consistent image acquisition settings and clipping [not over exposing any pixels at all] within the image for 'accurate' intensity measurements and capture in 12-bit [4096 grey levels, as any image analysis program can distinguish all those grey levels - we can only distinguish around 191 grey levels]. Plus you would have to be sure your image acquisition sampling procedure isn't adding any bias, as it's so tempting just to investigate the 'nicely labelled cells' that show 'good' colocalisation, whilst ignoring the rest [with the majority of the 'poorly labelled cells' being rejected as being a bit too typical] - although I guess you could analyse subsets of cell populations if your sampling is suitably randomised and you quantify the relative size of the subset population. However you must make sure any 'colocalisation' observed does not have any significant bleedthrough component [see my last post].

Regards

Keith

As usual there's a wealth on colocalisation [colocalization, co-localization] info online, e.g.
http://web.path.ox.ac.uk/~bioimaging//Stuff/Charges.html
In particular see the link to "Spatial quantitative analysis of fluorescently labeled nuclear structures: Problems, methods, pitfalls"

ImageJ and co-localisation apps:
http://www.uhnresearch.ca/facilities/wcif/imagej/colour_analysis.htm

Other example links
http://www.bitplane.com/go/products/imariscoloc?gclid=CLDJ483YnKICFQI9lAodbXfIuw
http://bioinformatics.oxfordjournals.org/cgi/reprint/21/15/3248
http://en.wikipedia.org/wiki/Colocalisation
http://www.olympusfluoview.com/applications/colocalization.html

and see the likes of the Zeiss Spectral separation & Emission fingerprinting [bleedthrough] guides at
http://www.well.ox.ac.uk/zeiss-confocal
if you suspect any bleedthrough component in the 'colocalisation'.



---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford OX3 7BN,
United Kingdom.

Telephone: +44 (0)1865 287568
Email: kjmorris-at-well.ox.ac.uk
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy

-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: 11 June 2010 09:22
To: kjmorris-at-well.ox.ac.uk

A word of caution about the part of the message from Keith dealing with quantification.
Be VERY careful with quantification in fluorescence. A higher signal doesn't necessarily mean a higher concentration of the antigen, it may also mean a better antibody, a better accessibility for the antibody, a better stability from the laser light and so many other things.
You could trace a line through the structure and show a profile scan (confocals are good at that), this is a nice way to concentrate the attention only on one structure and demonstrate the colocalization of 2 peaks of intensity. The profile also allows a fine analysis of the both signals intensities and who knows, perhaps you'll discover that both signals are not just perfectly superimposed, but are next to each other (very close), or that one signal has more a ring pattern and so on. Small differences in intensities are not easily appreciated with eyes (especially at high intensities), but when they are presented in histograms it is much easier to see.

It may be that both signals cannot be collected at a similar intensity as to observe a nice mix of both (like a nice yellow from red and green mixing). For profile scan it doesn't matter because you just want to show that the position of each peak is similar, whatever the height of the peaks. But for mixed images it is of course better if the structure has a well balanced mix of both signals (say the red is much stronger than the green, the merged image shows a red structure; well it is not a very convincing demonstration of colocalization).
Thus it is often required to increase the intensity of one signal. Take care to increase the overall intensity of the whole picture and not only part of the histogram (like playing with the contrast) or even worse: only a part of the image (this is a misconduct). If playing with the contrast is acceptable, do not forget to state it but I would avoid it.

Good luck
Â
Stephane

----- Original Message ----
X-from: "kjmorris-at-well.ox.ac.uk" {kjmorris-at-well.ox.ac.uk}
To: nizets2-at-yahoo.com
Sent: Thu, June 10, 2010 11:07:29 AM

Hi Molly

Just to add to Stephane's and Jerry's comments. You are right that a
confocal will not benefit from deconvolution software anywhere near as much
as a widefield microscope, for the reason you suggest, i.e. much of the
out-of focus information is removed by the confocal iris/pinhole in any
case. I use Improvison's [Perkin Elmer] 3D Volocity software to look at
things like 3D images, 3D cellular volumes and cell morphometry. In order to
get the best from a Z slice you are advised to over sample the z-stack, i.e.
for a 0.9um optical slice capture the z stack at least 0.45um apart [our
Zeiss 510 has a button to automatically 'double oversample' like this]. For
the likes of Volocity deconvolution, Improvision recommended to me that you
massively oversample the confocal z-stack with ten or more slices to every
0.9um z focus step [I think the rep suggested perhaps up to as high as 100x
oversampling]. I rarely use the optional 'Restoration' module
[deconvolution] Volocity module as I only use the confocal for z-stack, and
double oversampling and 3D reconstruction alone with Volocity is adequate
for my needs.

For co-localisation I would agree with Stephane though, and initially try
and tackle the co-localisation quantification in 2D using MetaMorph
[probably just because it's just easier]. I don't use ImageJ very much as I
have a MetaMorph licence, and we don't have the 3D Quantitation module for
Volocity in any case, that might be needed for 3D co-localisation
quantification - if interested just ask Improvision about that as their
support is superb.

Always be suspicious that any co-localisation isn't bleedthrough. You can
check this via spectral un-mixing on a 510 Metahead or just try imaging
single labeled samples of each fluorochrome and check there's no signal in
the other channels where there is now no longer any label [at the same
confocal imaging settings]. Don’t' forget a sample with no label just in
case of autofluorescence.

To quote Improvisions own website: "If you acquire your images from a
widefield microscope and want to volume render or make measurements from the
image data, you will need to use an image restoration technique
[de-convolution] to remove the out-of-focus information - a product of the
optical properties of the microscope. Even if your images are captured using
a laser scanning or spinning disk confocal microscope, the image quality may
benefit from image restoration." The word 'may' is probably significant.

Regards

Keith
---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford OX3 7BN,
United Kingdom.

Telephone:Â +44 (0)1865 287568
Email:Â kjmorris-at-well.ox.ac.uk
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy

-----Original Message-----
X-from: mshaw1-at-lumc.edu [mailto:mshaw1-at-lumc.edu]
Sent: 07 June 2010 14:37
To: kjmorris-at-well.ox.ac.uk

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://microscopy.com/MLFormMail.html
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Email: mshaw1-at-lumc.edu
Name: Molly Shaw

Organization: Loyola University Chicago

Title-Subject: [Filtered] deconvolution & confocal microscopy

Message: Hello,

I am a first-year graduate student trying to get started with
colocalization analysis of IF images taken by confocal microscopy. I
am working with fixed cells and mainly trying to measure
colocalization of 2 proteins. My main questions are concerning the
use of z-stacks and deconvolution:
1. Are z-stacks always necessary for measurement of colocalization,
or are single images sufficient?
2. I understand that deconvolution is essential for wide-field
images, but does it improve results that much when you're already
using confocal?
3. If you would recommend deconvolution, is the ImageJ plug-in a good
one to use?

Thank you for your time & help. I have read many articles on this
topic, but I keep ending up with conflicting answers.

Thanks again,
Molly Shaw

 Login Host: 67.174.84.137
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Hi listers,

In a turn of events that will make many of you smile knowingly, I've been asked to give a half day, hands-on "workshop" on "EM" as a professional development activity for some local high school science teachers. It's a great idea, but, as you all well know, there is just no such thing as throwing something on the SEM or TEM for a "quick pic"! Thankfully, we have an SEM with LV capabilities so that we can look at unfixed samples!

Some of these teachers are physics teachers and so probably not so excited about looking at the biological samples with which I am well-acquainted and bringing the micrographs back to their students. Do any of the materials scientists out there have a suggestion for an easy (and easily available) materials sample to look at with these teachers?

Thanks in advance for any advice that you can offer,
Kristen Lennon
Frostburg State University
Dept of Biological Sciences
Frostburg, MD 21532
kalennon-at-frostburg.edu
http://www.frostburg.edu/dept/biol/





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From: Frank_Karl-at-lincolnelectric.com
Date: Mon, 21 Jun 2010 12:34:02 -0500
Subject: [Microscopy] Re: SEM-Materials Demo

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

Kristen,

Several years ago a group from Dupont in Delaware gave a presentation at the Philadelphia Society for Microscopy, mostly biological focused. They showed SEM images of failure analysis that was of interest to all in that they could not show failure analysis of their work projects. Some that I remember were a screw that had to top twisted off and a broken flusher handle from a toilet. These showed force and corrosion. Perhaps you have something broken like these at home.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E - Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not represent the NIH. This message is not confidential and can be freely shared and reproduced.

________________________________
X-from: {kamlennon-at-yahoo.com}
Reply-To: {kamlennon-at-yahoo.com}

Hi Kristen,

2 low prep samples that show a nice fusion of biology and
physics/engineering principles are a butterfly wingscale and the
structure of a chicken eggshell. I have images on my website. Neither
requires critical point drying; just mount and sputter. The eggshell is
the surface exposed when the inner tough membrane layer is peeled off
(bet someone knows the official name....), but both sides are
interesting - outer surface shows the various fine CaCO3 beads that
scatter light so nicely...

The xylem image is a confocal one, but could be prepped for SEM. It is
made by finding the nice weed common here - Plantago _ and nicking the
petiole with a razor or fingernail and pulling apart - stripping out the
vascular bundles (like celery strings - that would do also...); these
wall thickenings support the xylem from collapsing due to atmospheric
pressure when there is greatly lowered pressure from transpiration in
the leaves (how much pressure does it take to get water to the top of a
redwood?)

http://www.bio.umass.edu/microscopy/gallery.htm


Hope this helps,

Dale

kamlennon-at-yahoo.com wrote:
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} Hi listers,
}
} In a turn of events that will make many of you smile knowingly, I've been asked to give a half day, hands-on "workshop" on "EM" as a professional development activity for some local high school science teachers. It's a great idea, but, as you all well know, there is just no such thing as throwing something on the SEM or TEM for a "quick pic"! Thankfully, we have an SEM with LV capabilities so that we can look at unfixed samples!
}
} Some of these teachers are physics teachers and so probably not so excited about looking at the biological samples with which I am well-acquainted and bringing the micrographs back to their students. Do any of the materials scientists out there have a suggestion for an easy (and easily available) materials sample to look at with these teachers?
}
} Thanks in advance for any advice that you can offer,
} Kristen Lennon
} Frostburg State University
} Dept of Biological Sciences
} Frostburg, MD 21532
} kalennon-at-frostburg.edu
} http://www.frostburg.edu/dept/biol/
}
}
}
}
}
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8, 22 -- From dac-at-research.umass.edu Mon Jun 21 11:41:10 2010
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From mine-at-tr.ibm.com Mon Jun 21 12:08:48 2010
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Microscope light bulb filaments are always of interested, especially burnt
out ones. Be careful cutting the glass envelope with a scriber and
breaking it. Ball point pen tips are interesting (make sure you clean the
ink out) as are tips of hypodermic needles, especial one with compound
bevels, metal savings (file a penny and nickel to met a mixed metal to demo
EDS if you have it).

What you need next is a little fictional story I which the investigator
solves some problem/crime/mystery with the sample you have in the scope. I
have always found the trick, it is a trick is to slant the story to involve
the audience.

Have fun...
Frank




kamlennon-at-yahoo.c
om
To
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Please respond to [Microscopy] SEM-Materials Demo
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Hi listers,

In a turn of events that will make many of you smile knowingly, I've been
asked to give a half day, hands-on "workshop" on "EM" as a professional
development activity for some local high school science teachers. It's a
great idea, but, as you all well know, there is just no such thing as
throwing something on the SEM or TEM for a "quick pic"! Thankfully, we have
an SEM with LV capabilities so that we can look at unfixed samples!

Some of these teachers are physics teachers and so probably not so excited
about looking at the biological samples with which I am well-acquainted and
bringing the micrographs back to their students. Do any of the materials
scientists out there have a suggestion for an easy (and easily available)
materials sample to look at with these teachers?

Thanks in advance for any advice that you can offer,
Kristen Lennon
Frostburg State University
Dept of Biological Sciences
Frostburg, MD 21532
kalennon-at-frostburg.edu
http://www.frostburg.edu/dept/biol/





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From: werner1-at-slb.com
Date: Mon, 21 Jun 2010 12:43:56 -0500
Subject: [Microscopy] SEM-Materials Demo

Contents Retrieved from Microscopy Listserver Archives
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Dr. Lennon,

According to your web site, your University has a Materials Engineering department. They probably have a tensile tester and may have a Charpy Impact tester. In any case, they have the capacity to break steel samples.

So ask for two pieces of the same steel, one fractured at room temperature and one at depressed (LN) temperature. People realize that things can get brittle at low temperature. The vivid difference between microvoid coalescence (overload) and cleavage (brittle) as seen in the SEM, at relatively low magnification, is something a Physics teacher or student will find immediately captivating.

And if they ask you why the steel behaves like that, just look smug and inform them that they need to attend classes to find out.

Regards,
Andrew Werner
Chief Metallurgist, Perforating
Schlumberger Reservoir Completions
14910 Airline Road
Rosharon, TX 77583

-----Original Message-----
X-from: kamlennon-at-yahoo.com [mailto:kamlennon-at-yahoo.com]
Sent: Monday, June 21, 2010 11:13 AM
To: Andrew Werner

Hi listers,

In a turn of events that will make many of you smile knowingly, I've been asked to give a half day, hands-on "workshop" on "EM" as a professional development activity for some local high school science teachers. It's a great idea, but, as you all well know, there is just no such thing as throwing something on the SEM or TEM for a "quick pic"! Thankfully, we have an SEM with LV capabilities so that we can look at unfixed samples!

Some of these teachers are physics teachers and so probably not so excited about looking at the biological samples with which I am well-acquainted and bringing the micrographs back to their students. Do any of the materials scientists out there have a suggestion for an easy (and easily available) materials sample to look at with these teachers?

Thanks in advance for any advice that you can offer,
Kristen Lennon
Frostburg State University
Dept of Biological Sciences
Frostburg, MD 21532
kalennon-at-frostburg.edu
http://www.frostburg.edu/dept/biol/





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From: DusevichV-at-umkc.edu
Date: Mon, 21 Jun 2010 12:54:55 -0500
Subject: [Microscopy] RE: viaWWW: SEM of air sensitive samples

Contents Retrieved from Microscopy Listserver Archives
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Somewhere I saw people using air-tight box with a
cover that was kept in place by suction (atmospheric pressure),
so that specimen in the box was kept under vacuum.
Small spring, attached to the cover, was used to move it away
when air pressure in a SEM specimen chamber was low enough.
Specimen was conductive and the box was made from metal.

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

} -----Original Message-----
} From: jacqueline.ayotte-at-ticona.com
} [mailto:jacqueline.ayotte-at-ticona.com]
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} Subject: [Microscopy] viaWWW: SEM of air sensitive samples
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} Email: jacqueline.ayotte-at-ticona.com
} Name: Jackie Ayotte
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} Organization: Ticona Engineering Polymers
}
} Title-Subject: [Filtered] SEM of air sensitive samples
}
} Message: Hello All,
}
} I am challenged with the task of performing SEM on air sensitive
} catalysts. Can anyone share their method for doing this? I have seen
} SEM images of the catalysts that I am interested in analyzing.
} However the company that generated those micrographs can not share
} their procedure with me.
}
} I can provide more information regarding the catalyst off-line.
}
} Much Thanks -
}
} Jackie
}
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5, 25 -- From DusevichV-at-umkc.edu Mon Jun 21 12:54:54 2010
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From: swalck-at-southbaytech.com
Date: Mon, 21 Jun 2010 13:38:28 -0500
Subject: [Microscopy] viaWWW: SEM of air sensitive samples

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

Jacqueline,
I have recommended the following procedure to people using our
SampleSaver(TM) storage containers. These containers are compatible with
glove boxes, so if you are using a glove box, you simply close your sample
up in the container without the need to purge the unit.

The SampleSaver(TM) container is used to transport the sample from wherever
you are processing/preparing the sample under shielded conditions to the
microscope. There are glove bags with large open ends that you can place
the unopened SampleSaver(TM) container in and then tie the glove bag over
the open end of the SEM. You then can fill the glove bag with nitrogen and
then take the sample out of the SampleSaver(TM) container in the glove bag,
place the sample in the SEM stage, close the stage and pump it out. I
would, of course, bring the SEM chamber up to atmosphere with nitrogen and
purge it while the glove bag is on prior to opening the SampleSaver(TM).

Disclaimer: South Bay Technology, Inc. manufactures and sells the
SampleSaver(TM) Storage Container. We do not sell the glove bag.

-Scott
 
Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA  92673
 
US Toll Free: 1-800-728-2233
Tel: (949) 492-2600
Fax: (949) 492-1499
 
www.southbaytech.com
swalck-at-southbaytech.com

-----Original Message-----
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Sent: Friday, June 18, 2010 11:50 PM
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Email: jacqueline.ayotte-at-ticona.com
Name: Jackie Ayotte

Organization: Ticona Engineering Polymers

Title-Subject: [Filtered] SEM of air sensitive samples

Message: Hello All,

I am challenged with the task of performing SEM on air sensitive
catalysts. Can anyone share their method for doing this? I have seen
SEM images of the catalysts that I am interested in analyzing.
However the company that generated those micrographs can not share
their procedure with me.

I can provide more information regarding the catalyst off-line.

Much Thanks -

Jackie

Login Host: 148.163.178.12
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From: RossLM-at-missouri.edu
Date: Mon, 21 Jun 2010 13:39:27 -0500
Subject: [Microscopy] EM Research Lab Technician immediate opening at the Univ of Missouri

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

The Electron Microscopy Core (EMC) at the University of Missouri in Columbia, MO has an opening for an entry-level electron microscopy technician. The EMC is a campus-wide service facility housing 2 SEM's (Hitachi S-4700 and FEI Quanta 600F) and a JEOL 1400 TEM. Additionally, a new HVTEM will be installed in the next year. More about the lab can be found at: http://www.emc.missouri.edu/.

The primary responsibilities of the new staff member would be in the area of biological and materials sample preparation for both SEM and TEM. Duties include basic and advanced sample preparation techniques as well as the development of new and specialized techniques for unique client applications. We are seeking an individual that is organized, meticulous, creative and service oriented. Laboratory experience is required, although not necessarily EM lab experience. For more information and to apply for the Research Lab Technician position, go to https://jobs.missouri.edu/vacdetails.php?vac=1017122.

Lou Ross
--
Sr. Research Specialist
University of Missouri
Electron Microscopy Core Facility
W136 Veterinary Medicine Building
Columbia, MO 65211
573.882.4777, fax 573.884.2227
RossLM-at-missouri.edu
http://www.emc.missouri.edu/



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From: DusevichV-at-umkc.edu
Date: Mon, 21 Jun 2010 17:51:54 -0500
Subject: [Microscopy] RE: SEM-Materials Demo

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


On Jun 21, 2010, at 9:12 AM, kamlennon-at-yahoo.com wrote:

} Hi listers,
}
} In a turn of events that will make many of you smile knowingly, I've
} been asked to give a half day, hands-on "workshop" on "EM" as a
} professional development activity for some local high school science
} teachers. It's a great idea, but, as you all well know, there is
} just no such thing as throwing something on the SEM or TEM for a
} "quick pic"! Thankfully, we have an SEM with LV capabilities so that
} we can look at unfixed samples!
}
} Some of these teachers are physics teachers and so probably not so
} excited about looking at the biological samples with which I am well-
} acquainted and bringing the micrographs back to their students. Do
} any of the materials scientists out there have a suggestion for an
} easy (and easily available) materials sample to look at with these
} teachers?
}
} Thanks in advance for any advice that you can offer,
} Kristen Lennon
} Frostburg State University
} Dept of Biological Sciences
} Frostburg, MD 21532
} kalennon-at-frostburg.edu
} http://www.frostburg.edu/dept/biol/


Dear Kristen,
I am not a materials person, but I did have a project looking at
sediment from river bottoms. The samples are very easy to prepare--
just suspend the mud in a fairly dilute suspension, let the larger
particles settle, then put a drop (a few ul will do) onto a carbon/
formvar coated grid and let dry. If my experience was typical, you
will see diatom skeletons, mineral fragments, crystalline and not, and
other items in a TEM image. EDS (if available) will also show some
surprises for some of the particles. I saw U and Au in addition to
the usual suspects, Na, K, Si, Al, Ti, Ca, etc. Good luck.
Yours,
Bill
PS. I am no longer working for FEI.

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From kefxenaholicdiz-at-xenaholic.com Mon Jun 21 15:59:58 2010
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Kristen,

You can use paper. Place on the stub two pieces of paper: writing
paper and filter paper. You can see paper fibers and filler particles in
writing paper (no particles in filter paper). You can check distribution
of filler with BSE and composition with EDS (usually some salts of Ca).

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784


} -----Original Message-----
} From: kamlennon-at-yahoo.com [mailto:kamlennon-at-yahoo.com]
} Sent: Monday, June 21, 2010 11:03 AM
} To: Dusevich, Vladimir
} Subject: [Microscopy] SEM-Materials Demo
}
}
}
}
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}
} Hi listers,
}
} In a turn of events that will make many of you smile knowingly, I've
} been asked to give a half day, hands-on "workshop" on "EM" as a
} professional development activity for some local high school science
} teachers. It's a great idea, but, as you all well know, there is just
} no such thing as throwing something on the SEM or TEM for a "quick
} pic"! Thankfully, we have an SEM with LV capabilities so that we can
} look at unfixed samples!
}
} Some of these teachers are physics teachers and so probably not so
} excited about looking at the biological samples with which I am well-
} acquainted and bringing the micrographs back to their students. Do any
} of the materials scientists out there have a suggestion for an easy
} (and easily available) materials sample to look at with these teachers?
}
} Thanks in advance for any advice that you can offer,
} Kristen Lennon
} Frostburg State University
} Dept of Biological Sciences
} Frostburg, MD 21532
} kalennon-at-frostburg.edu
} http://www.frostburg.edu/dept/biol/
}
}
}
}
}
} ==============================Original
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7, 25 -- From DusevichV-at-umkc.edu Mon Jun 21 17:51:54 2010
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From: oshel1pe-at-cmich.edu
Date: Tue, 22 Jun 2010 07:16:38 -0500
Subject: [Microscopy] Demo sample

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

I encountered the same situation on a number of occasions over the
years I was teaching electron microscopy. I am reluctant to admit
it, but the SEM sample that got the greatest attention from high
school students was an ant, and next best (and more difficult to
prepare) was a mosquito. I hate to admit it, but beautifully etched
specimens of pearlitic steel mostly left them cold.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From ghenry-at-ghowardassociates.com Mon Jun 21 23:47:28 2010
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When I was at PPG's Glass Technology Center, we had an open house for family
members. We did a demo on the SEM that was kind of neat. We set them up
for a punch line at the end of the demo. Here's what we did:

We took a dead fly and heavily coated it with gold. It was a really thick
layer of gold. This was done prior to the demonstration. Our real sample
was a cross section of enamel on glass that we wanted to show the layers and
components and measure the different phases with the XEDS to show how we
measure composition.

To introduce them to the SEM, we showed them the fly. Scanned it all over
and explained how the image was formed. We did not tell them it was coated.
Of course, they thought that it was neat.

Then we moved to the cross section sample and showed how we measured the
composition of the different phases with the X-ray system. Oh-hum, kind of
boring.

Then we asked them "what was the composition of the fly?" Some would say
C,H,O or whatever their guesses were. We then said, "Well, let's find out
for sure." And, of course, the only thing that came up was gold in the
spectrum. We then asked, "Why is it gold?" and then let them guess. At the
end, if they didn't know, we asked them if we could interest them in stock
certificates in the "PPG Lost Gold Bug Mine" which were certificates that we
made up with a big yellow faux colored SEM image of the fly, the X-ray
spectrum in the background, and the signed signatures of our group members
as officers in the "PPG Lost Gold Bug Mine Corporation" . I think that it
went over fairly well and the kids were carrying around their stock
certificates all day.

-Scott
 
Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA  92673
 
US Toll Free: 1-800-728-2233
Tel: (949) 492-2600
Fax: (949) 492-1499
 
www.southbaytech.com
swalck-at-southbaytech.com




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From internet-at-euroforum.com Tue Jun 22 03:10:27 2010
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Hi
When demonstrating to 6th formers I've asked for volunteers to provide
hair samples, stuck them on a stub & imaged them (in esem uncoated) Try
& get a variety of hair types to see differences.
Good luck with the demo
Nikki
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Hmm, fun. I run a couple of "Grandparents
University" sessions here in the summer
(tomorrow) -- grandparents and their grandkids.
This would be fun, but the cool purple glow of
the sputter coater always gets the kids excited
for the rest of the session. Which is "bring a
bug, and we'll look at it in the SEM". A big
favorite.
I've thought of sawn/cut metal vs brittle
fracture in LN2, but really, looking at bugs they
caught gets the kids the most excited, and that's
where all our obsessions started, yes?

Phil

} When I was at PPG's Glass Technology Center, we had an open house for family
} members. We did a demo on the SEM that was kind of neat. We set them up
} for a punch line at the end of the demo. Here's what we did:
}
} We took a dead fly and heavily coated it with gold. It was a really thick
} layer of gold. This was done prior to the demonstration. Our real sample
} was a cross section of enamel on glass that we wanted to show the layers and
} components and measure the different phases with the XEDS to show how we
} measure composition.
}
} To introduce them to the SEM, we showed them the fly. Scanned it all over
} and explained how the image was formed. We did not tell them it was coated.
} Of course, they thought that it was neat.
}
} Then we moved to the cross section sample and showed how we measured the
} composition of the different phases with the X-ray system. Oh-hum, kind of
} boring.
}
} Then we asked them "what was the composition of the fly?" Some would say
} C,H,O or whatever their guesses were. We then said, "Well, let's find out
} for sure." And, of course, the only thing that came up was gold in the
} spectrum. We then asked, "Why is it gold?" and then let them guess. At the
} end, if they didn't know, we asked them if we could interest them in stock
} certificates in the "PPG Lost Gold Bug Mine" which were certificates that we
} made up with a big yellow faux colored SEM image of the fly, the X-ray
} spectrum in the background, and the signed signatures of our group members
} as officers in the "PPG Lost Gold Bug Mine Corporation" . I think that it
} went over fairly well and the kids were carrying around their stock
} certificates all day.
}
} -Scott
} Ý
} Scott D. Walck, Ph.D.
} Technical Director
} South Bay Technology, Inc.
} 1120 Via Callejon
} San Clemente, CAÝ 92673
} Ý
} US Toll Free: 1-800-728-2233
} Tel: (949) 492-2600
} Fax: (949) 492-1499
} Ý
} www.southbaytech.com
} swalck-at-southbaytech.com

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576


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From: tina-at-pbrc.hawaii.edu
Date: Tue, 22 Jun 2010 21:49:19 -0500
Subject: [Microscopy] Laser capture microdissection

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

Bugs are always a big hit, but one set of samples I usually get a lot of
interest in from kids of all ages is record grooves vs. pits on a CD.
Usually the older generation gets involved describing to their
grandchildren what exactly a record was, and a lively discussion of
analog versus digital music develops. Seeing one at relatively low
magnification vs the other at high mag really drives home the advance of
technology. Having a computer chip in the scope at the same time usually
goes over well. The hassle of dissolving the CD and burning off the chip
packaging is well worth it, and once prepared, they can last for years.

Oh, there's also this: take the back off of an old (but working)
mechanical wristwatch. Watching the spring and gears move at TV rates is
oddly hypnotic. Dropping into slow scan mode also works into the
explanation of how the image is formed.

Over the years, I seem to get the best reaction from microscopical
novices if they look at things they have some sort of connection with at
a scale they are used to in normal life.

Hope this helps,

Jim

--

James M. Ehrman
Digital Microscopy Facility
Mount Allison University
63B York St.
Sackville, NB E4L 1G7
CANADA

phone: 506-364-2519
fax: 506-364-2505
email: jehrman-at-mta.ca
www: http://www.mta.ca/dmf

For people who like peace and quiet
- a phoneless cord.


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Hi,



We need set up the whole TEM sample preparation lab mainly for thin films specimen. I heard that Gantan PIPS can do ion milling in the last stage. Where can I buy used PIPS or else like PIPS? Please give some suggestion in setting up a TEM sample preparation lab involving thin films samples, materials samples, protein samples, et al. Thank you very much.



Fengli



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From international-at-sunderland.ac.uk Tue Jun 22 11:23:46 2010
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Folks,
Please note that today the M&M2010 EXPO that describes the program of the meeting is available on-line at:
http://journals.cambridge.org/action/displayIssue?jid=MAM&volumeId=16&issueId=S1&iid=7816538

Like the rest of our journal it is searchable, but the search engine is not too intuitive. If you are looking for presentations by colleagues and friends, I suggest that you download the "Scientific Program" and search that with Acrobat or similar.


http://journals.cambridge.org/action/displayFulltext?type=1&pdftype=1&fid=7816585&jid=MAM&volumeId=16&issueId=S1&aid=7816584

--
John Mansfield PhD Cphys CSci MInstP
2010 Microscopy & Microanalysis Program Chair
North Campus Electron Microbeam Analysis Laboratory
417 SRB, University of Michigan
2455 Hayward, Ann Arbor MI 48109-2143 USA
Phone: (734) 936-3352 FAX (734) 763-2282
Cell. Phone: (734) 834-3913
(Leaving a phone message at 936-3352 is preferable to 834-3913)
Email: jfmjfm-at-umich.edu
URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
Location: N 42 ° 17' 38.3" (42 ° 17.6391' ) W 83 ° 42' 38.6" (-83 ° 42.6439')
AIM: thejfmjfm
Yahoo: thejfmjfm
Skype: thejfmjfm

Home address:
4304 Spring Lake Boulevard
Ann Arbor MI 48108-9657
Phone (734) 994-3096
Location: N 42 ° 13' 28.8" (42 ° 13.4807') W 83 ° 45' 47.9" 42° 16' 48" (-83 ° 45.7980')

Please note: Electronic Mail is not secure, but should be read several times every day, and should definitely be used for urgent or sensitive issues.






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Hi, All-

Someone bought a Zeiss PALM laser capture microdissection and laser
tweezers system and put it in our core facility, so now I'm "in charge" of
it. Unfortunately, I have zero experience, so I can't even begin to
predict what problems I will run into. The first one is (after I figured
out how to turn it on) that the user wants to capture diatoms with
their attendant microbes for PCR. We can't see the diatoms in droplets on
the membrane slides, which are hydrophobic, because the (high salt) media
beads up like mad. If we dry it down, all we see is salt. If we wash the
beasties too much, they rupture and the microbes fall off.

Can any of you offer advice and/or point me to resources for this
microscope system? I know they're out there, but I'd like to find someone
with experience with marine microbes or similar.

Aloha,
Tina

http://www5.pbrc.hawaii.edu/bemf/site/
http://www5.pbrc.hawaii.edu/microangela/

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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From: Peter.Steele-at-allkids.org
Date: Wed, 23 Jun 2010 00:37:58 -0500
Subject: [Microscopy] viaWWW: Ultracut S ultramicrotome controller

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Email: Peter.Steele-at-allkids.org
Name: P. Steele

Organization: All Children's Hospital

Title-Subject: [Filtered] Ultracut S ultramicrotome controller.

Message: I am looking for the Controller for a
Reichert Ultracut S ultramicrotome. The machine
is approximately 20 years old and Leica reports
parts are no longer available for this workhorse.
This is our backup ultramicrotome. The controller
is defective but the ultramicrotome is in good
shape.


If you have an old Ultracut S ultramicrotome
Controller that you are would like to donate
please contact me offline.

Thank you.


Peter O. Steele, PhD, PMIAC, CLDir
Pathology and Laboratory Medicine
All Children's Hospital
St. Petersburg, FL,
USA, 33731-8920

voice: 727 767-4465
beeper: 727 825-5131


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From: R.G.Phillips-at-sussex.ac.uk
Date: Wed, 23 Jun 2010 05:53:11 -0500
Subject: [Microscopy] MRC Studentships in Imaging MSc at University of Sussex

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Email: innap-at-savion.huji.ac.il
Name: Inna Popov

Organization: The Hebrew University of Jerusalem

Title-Subject: [Filtered] SEM Materials - Demo

Message: Hi,
Once in the past I used to demonstrate SEM to 7 years old children.
Observation of a coin had attracted their attention. But, to be
honest they were too young.
Later I took colledge students to SEM&EDS lesson and used two metal
soliers from chocolate egg
( see the link:
http://www.soldatiki.ru/2008/soldatiki.php?page=10&lang=en). Such
objects attracts a lot of interest, because they have a story behind
(ancient art of metallurgy, history of Roman Empire - depends on what
you show). They story is a key point which will remain in the memory
of pupils. And you simultaneously demonstrate 1)typical defects of
casting; 2) chemical composition; 3)brittle fracture of casting (no
problem to break some tiny part of a soldier) and whatever you wish
to add.
Alternatively, you may take a plastic toy from a KinderJoy egg, use
LV mode, show magnified SE image of the object, explain why they do
not see colors, how to reconstruct a preparation way, i.e. story.
The last attractive thing I may imagine is simply corrosion deposite.
A day before the lesson put a drop of saline on a piece of steel
(magnesium alloy is better). At the lesson you will demonstrate
beautiful aestetic oxide-hydroxide flower-like flakes and give a
story.
Good luck,
Inna

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From jadams-at-nw.hidta.org Wed Jun 23 01:57:54 2010
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The Medical Research Council is offering five fully funded studentships for
study commencing October 2010 in the University of Sussex Master's Degree
programme in Imaging in Biomedical Research. This is a one year 'taught'
programme in the Biochemistry Department. It includes lecture, practical and
individual research components designed to introduce the student to a broad
range of biological and medical imaging techniques and to develop advanced
skills in image acquisition, analysis and applications in biomedical
research.
UK applicants are eligible for fees plus living allowance award
while other EU applicants are eligible for fees award only.
Your application must be submitted by 16th July 2010 to be considered for
these studentships.

For further information see our prospectus:
http://www.sussex.ac.uk/study/pg/subjects/areasofstudy/Biochemistry/22196
and our postgraduate application system:
https://www.sussex.ac.uk/pgapplication/.
For additional details about the programme and the Sussex Centre for
Advanced microscopy refer to our website at:
http://www.sussex.ac.uk/lifesci/1-4-30.html.


Dr Roger Guy Phillips
Centre for Advanced Microscopy,
University of Sussex
School of Life Sciences
John Maynard Smith Building
Falmer, Brighton & Hove
BN1 9QG
United Kingdom

phone:44 (0)1273 877585
fax: 44 (0)1273 678433
email: R.G.Phillips-at-sussex.ac.uk
room:2C9 (ext 7585)/lab 4C2 (ext 2734)



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From: milesd-at-us.ibm.com
Date:
Subject:

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

Peter,

If you don't get a donation and want to get it repaired, I have been
trying to connect people with aftermarket service for these older
microtomes and have a web page with some resources:

http://people.umass.edu/dac/projects/Open_Cut/Ultracut_Service.html

I recommend contacting MOC; Helmut Patzig formerly worked for Leica. He
knows people in Austria and has told me they can still supply the
controller boards - they aren't cheap, but neither is a new microtome.

I also have an "S" with dead controller. The main circuit board has
three separate microprocessors and memories of a now-obsolete types. The
functionality can today be provided by a $3 micro-controller; if only I
had more time....

Cheers!

Dale


Peter.Steele-at-allkids.org wrote:
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} Title-Subject: [Filtered] Ultracut S ultramicrotome controller.
}
} Message: I am looking for the Controller for a
} Reichert Ultracut S ultramicrotome. The machine
} is approximately 20 years old and Leica reports
} parts are no longer available for this workhorse.
} This is our backup ultramicrotome. The controller
} is defective but the ultramicrotome is in good
} shape.
}
}
} If you have an old Ultracut S ultramicrotome
} Controller that you are would like to donate
} please contact me offline.
}
} Thank you.
}
}
} Peter O. Steele, PhD, PMIAC, CLDir
} Pathology and Laboratory Medicine
} All Children's Hospital
} St. Petersburg, FL,
} USA, 33731-8920
}
} voice: 727 767-4465
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} Floridaís west coast. Founded in 1926, All
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==============================Original Headers==============================
9, 20 -- From dac-at-research.umass.edu Wed Jun 23 06:38:34 2010
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9, 20 -- Date: Wed, 23 Jun 2010 07:39:30 -0400
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From matt-at-jaysseptic.com Wed Jun 23 07:28:00 2010
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Hi all!

I am looking for protocols to label bacterial biofilm for detection in SEM/EDX.
We are examining irregular surfaces and the characterization of the biofilm only by morphology is not easy.
Given that biofilms are composed of polysaccharides, I wondered if it could not just be labeled with iodine (like lugol) ? It would be perfect because the substrate contains absolutely no trace of iodine, however we would probably not be able to make the difference between the bacteria themselves and the biofilm.
Any experience or food for thought to share?

Stephane




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From ghuman-at-add.mb00.net Wed Jun 23 10:23:47 2010
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Peter, and all.

Reading this post shook loose an old memory of a company I once located
that specializes in repairs of electronic circuit boards and equipment.
Part of their specialty is repairing obsolete electronics. I found them
when I had an old, no longer supported test system. I ended up not using
them (didn't repair system), so I can't vouch for them to that extent, but
they are BBB accredited, give free evaluations, and warranty their work
for one year. They are ACS Industrial, located in Maryland, USA, and
their web site is:

http://www.acsindustrial.com/index.php

I realized this could be useful information to others, keeping vintage,
unsupported equipment alive. Their site shows some nasty looking circuits
that they have repaired.

Good luck,
Darrell





X-from:
Peter.Steele-at-allkids.org
To:
Darrell Miles/Fishkill/IBM-at-IBMUS






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Email: Peter.Steele-at-allkids.org
Name: P. Steele

Organization: All Children's Hospital

Title-Subject: [Filtered] Ultracut S ultramicrotome controller.

Message: I am looking for the Controller for a
Reichert Ultracut S ultramicrotome. The machine
is approximately 20 years old and Leica reports
parts are no longer available for this workhorse.
This is our backup ultramicrotome. The controller
is defective but the ultramicrotome is in good
shape.


If you have an old Ultracut S ultramicrotome
Controller that you are would like to donate
please contact me offline.

Thank you.


Peter O. Steele, PhD, PMIAC, CLDir
Pathology and Laboratory Medicine
All Children's Hospital
St. Petersburg, FL,
USA, 33731-8920

voice: 727 767-4465
beeper: 727 825-5131


Confidentiality Notice: This e-mail message,
including any attachments, is for the sole use of
intended recipient(s) and may contain
confidential and privileged information. Any
unauthorized review, use, disclosure or
distribution is prohibited. If you are not the
intended recipient, please contact the sender by
reply e-mail and destroy all copies of the
original message.

Note: All Childrenís Hospital is the only
specialty licensed childrenís hospital on
Floridaís west coast. Founded in 1926, All
Childrenís has grown into a leading pediatric
referral center that is dedicated to advancing
treatment, education, research and advocacy in
child health. See www.allkids.org .


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From: vwporsche-at-verizon.net
Date: Wed, 23 Jun 2010 11:16:38 -0500
Subject: [Microscopy] viaWWW: Service on JSM 6100

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From: AJBowling-at-dow.com
Date: Wed, 23 Jun 2010 12:05:53 -0500
Subject: [Microscopy] Biofilm detection

Contents Retrieved from Microscopy Listserver Archives
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Here are some ideas for you:

You might get something useful with osmium vapor (or maybe OTO), if your
substrate doesn't also react with it.

Maybe do a PATAg and look for silver deposition on the polysaccharides
of the biofilm.

Another possibility is that there might be a polysaccharide-degrading
enzyme that can degrade the polysaccharides of the biofilm. If so, that
enzyme could be conjugated to colloidal gold and then used to probe your
sample.

Good luck,

Andy



-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Wednesday, June 23, 2010 9:32 AM
To: Bowling, Andrew (AJ)

Hi all!

I am looking for protocols to label bacterial biofilm for detection in
SEM/EDX.
We are examining irregular surfaces and the characterization of the
biofilm only by morphology is not easy.
Given that biofilms are composed of polysaccharides, I wondered if it
could not just be labeled with iodine (like lugol) ? It would be perfect
because the substrate contains absolutely no trace of iodine, however we
would probably not be able to make the difference between the bacteria
themselves and the biofilm.
Any experience or food for thought to share?

Stephane




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From: rstiger-at-ppg.com
Date: Thu, 24 Jun 2010 14:40:01 -0500
Subject: [Microscopy] viaWWW: Optical Microscopy: Phase, Nomarski

Contents Retrieved from Microscopy Listserver Archives
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Jim,
The preparation of a CD is quite simple. You don't have to burn off the
plastic at all. Take a commercially prepared CD, not a recordable one, and
cut a square in the top label side of the CD with a razor blade. Then take
Scotch® tape and firmly press it onto the area that you scored with the
razor blade. When you remove the tape, the recording will come up and you
can put it in the microscope.

-Scott
 
Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA  92673
 
US Toll Free: 1-800-728-2233
Tel: (949) 492-2600
Fax: (949) 492-1499
 
www.southbaytech.com
swalck-at-southbaytech.com

-----Original Message-----
X-from: jehrman-at-mta.ca [mailto:jehrman-at-mta.ca]
Sent: Tuesday, June 22, 2010 6:03 AM
To: swalck-at-southbaytech.com

Bugs are always a big hit, but one set of samples I usually get a lot of
interest in from kids of all ages is record grooves vs. pits on a CD.
Usually the older generation gets involved describing to their
grandchildren what exactly a record was, and a lively discussion of
analog versus digital music develops. Seeing one at relatively low
magnification vs the other at high mag really drives home the advance of
technology. Having a computer chip in the scope at the same time usually
goes over well. The hassle of dissolving the CD and burning off the chip
packaging is well worth it, and once prepared, they can last for years.

Oh, there's also this: take the back off of an old (but working)
mechanical wristwatch. Watching the spring and gears move at TV rates is
oddly hypnotic. Dropping into slow scan mode also works into the
explanation of how the image is formed.

Over the years, I seem to get the best reaction from microscopical
novices if they look at things they have some sort of connection with at
a scale they are used to in normal life.

Hope this helps,

Jim

--

James M. Ehrman
Digital Microscopy Facility
Mount Allison University
63B York St.
Sackville, NB E4L 1G7
CANADA

phone: 506-364-2519
fax: 506-364-2505
email: jehrman-at-mta.ca
www: http://www.mta.ca/dmf

For people who like peace and quiet
- a phoneless cord.


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From jacky-at-gdmit.net Wed Jun 23 14:51:59 2010
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For any kind of student demo, you want to have a "hook" to engage the
students. We used a "CSI" theme and used light bulb filaments as our
samples. Our "story" was about a car accident where there was a
disagreement about whether the driver had his headlights on at the time
of the accident. The driver claimed his headlights were on and the
other person claimed that the driver turned on his lights after the
accident to make it LOOK like they were on. I showed 2 burned-out light
bulb filaments; one of which was burning when I broke the glass, the
other of which I broke the glass envelope and and then excited the
filament.

The filament which was hot when broken had glass embedded in the
filament wire, while the other was just oxidized. The glass globules
were quite visible embedded in the tungsten wire and I could then use
EDX to identify the particles. In the ensuing discussion, I had the
students deduce the actual scenario.

Note: be careful on breaking the bulbs. Glass can go flying
everywhere. I put mine in a paper bag and then used a vise to slowly
crack the glass envelope (so I didn't damage the filament).

Cheers,
Henk

At 6/21/2010 12:03 PM, kamlennon-at-yahoo.com wrote:
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} Hi listers,
}
} In a turn of events that will make many of you smile knowingly, I've been asked to give a half day, hands-on "workshop" on "EM" as a professional development activity for some local high school science teachers. It's a great idea, but, as you all well know, there is just no such thing as throwing something on the SEM or TEM for a "quick pic"! Thankfully, we have an SEM with LV capabilities so that we can look at unfixed samples!
}
} Some of these teachers are physics teachers and so probably not so excited about looking at the biological samples with which I am well-acquainted and bringing the micrographs back to their students. Do any of the materials scientists out there have a suggestion for an easy (and easily available) materials sample to look at with these teachers?
}
} Thanks in advance for any advice that you can offer,
} Kristen Lennon
} Frostburg State University
} Dept of Biological Sciences
} Frostburg, MD 21532
} kalennon-at-frostburg.edu
} http://www.frostburg.edu/dept/biol/
}
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--
Hendrik O. Colijn
www.ceof.ohio-state.edu
OSU Campus Electron Optics Facility colijn.1-at-osu.edu
040 Fontana Labs (614) 292-0674 (V)
116 W. 19th Ave. (614) 292-7523 (F)
Columbus, OH 43210

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

==============================Original Headers==============================
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Dear listers,
Below is the announcement for a postdoctoral position to start October 1
(negotiable). Please forward to any potentially interested parties.
Albina Borisevich
--

*Combined Scanning Transmission and Scanning Probe Microscopy of Oxides*

*Materials Science and Technology Division *

*Oak Ridge National Laboratory *

*Oak Ridge, Tennessee *

* *

*Project Description: *

The Materials Science and Technology Division at Oak Ridge National
Laboratory (ORNL) is seeking a candidate to fill a postdoctoral position
in the field of combined scanning transmission electron microscopy and
scanning probe microscopy of oxide thin films and nanostructures. The
position is available October 1 2010. This program takes advantage of
ORNL’s suite of advanced electron microscopes, including 5
aberration-corrected instruments, as well as (S)TEM/STM and (S)TEM/AFM
capabilities.

The successful applicant must demonstrate experience in electron
microscope operation, preferably FEI microscopes, as well as skills in
analysis and interpretation of microscopic and spectroscopic data. This
position provides an opportunity to join an experienced team working in
a highly collaborative environment. Interactions with the scanning probe
microscopy program at ORNL’s Center for Nanophase Materials Sciences
(CNMS) are anticipated on a daily basis.

*Qualifications: PhD degree required *

This position requires a Ph.D. in Materials Science, Physics, or related
field, with an emphasis on advanced TEM or STEM. Knowledge of oxide
crystal chemistry is a plus. Excellent oral and written communication
skills are required, and presentations and publication of scientific
results in peer-reviewed journals are expected. The applicant must have
the ability to work in a team and interact effectively with a broad
range of colleagues. Applicants cannot have received the most recent
degree more than five years prior to the date of application and must
complete all degree requirements before starting their appointment.

*Interested applicants please write to* to Drs. Albina Y. Borisevich,
albinab-at-ornl.gov {mailto:albinab-at-ornl.gov} , Stephen J. Pennycook,
pennycooksj-at-ornl.gov {mailto:pennycooksj-at-ornl.gov} , or Sergei V.
Kalinin, sergei2-at-ornl.gov {mailto:sergei2-at-ornl.gov} . This appointment is
offered through the ORNL Postgraduate Research Participation Program and
is administered by the Oak Ridge Institute for Science and Education
(ORISE). The program is open to all qualified U.S. and non-U.S. citizens
without regard to race, color, age, religion, sex, national origin,
physical or mental disability, or status as a Vietnam-era veteran or
disabled veteran.

--
Albina Y. Borisevich
R&D Staff
Electron Microscopy Group
Oak Ridge National Laboratory
Materials Science and Technology Division
PO Box 2008
Oak Ridge TN 37831-6031

http://stem.ornl.gov/

For express mail add: 1 Bethel Valley Road
phone: (865) 576-4060
fax: (865) 574-4143

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Email: rstiger-at-ppg.com
Name: Rebecca Stiger

Organization: PPG

Title-Subject: [Filtered] Optical Microscopy: Phase, Nomarski

Message: We're in a situation where we could really use some training
in theory and practice of Nomarski and Phase Contrast optical
imaging. I've been in touch with McCrone, but they seem to focus on
Polarized Light Microscopy and it's applications.

Any suggestions on good training for Nomarski and/or Phase? It would
be nice to get all three techniques bundled into one event, but I
don't know if it's possible.

Many thanks,

Becca



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From: bozzola-at-siu.edu
Date: Thu, 24 Jun 2010 14:57:49 -0500
Subject: [Microscopy] SEM: Spectral Engine II Card

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

A colleague of mine needs a 4pi Spectral Engine II card for a SEM
system in his lab.

Does anyone have such a used, working card for sale?

Thank you.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

==============================Original Headers==============================
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From: baskin-at-bio.umass.edu
Date: Thu, 24 Jun 2010 17:08:53 -0500
Subject: [Microscopy] Re: viaWWW: Optical Microscopy: Phase, Nomarski

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

Becca,
You did not indicate where PPG is. But if you are near a
university then look for someone in a biology department who teaches
microscopy or does a bunch of light microscopy (check their web
pages) and they might be able to help. Depends a little on how much
information you are after.

Good luck,
Tobias




} Email: rstiger-at-ppg.com
} Name: Rebecca Stiger
}
} Organization: PPG
}
} Title-Subject: [Filtered] Optical Microscopy: Phase, Nomarski
}
} Message: We're in a situation where we could really use some training
} in theory and practice of Nomarski and Phase Contrast optical
} imaging. I've been in touch with McCrone, but they seem to focus on
} Polarized Light Microscopy and it's applications.
}
} Any suggestions on good training for Nomarski and/or Phase? It would
} be nice to get all three techniques bundled into one event, but I
} don't know if it's possible.
}
} Many thanks,
}
} Becca
}
}
}
} Login Host: 141.189.251.1
} ---------------------------------------------------------------------------
}

--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Biology Department
/_ / __ /__ \ \ \__ 611 N. Pleasant St.
/ / / \ \ \ University of Massachusetts
/ / / \ \ \ Amherst, MA, 01003
/ / ___ / \ \__/ \ ____
www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

==============================Original Headers==============================
7, 21 -- From baskin-at-bio.umass.edu Thu Jun 24 17:08:52 2010
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7, 21 -- To: rstiger-at-ppg.com
7, 21 -- From: Tobias Baskin {baskin-at-bio.umass.edu}
7, 21 -- Subject: Re: [Microscopy] viaWWW: Optical Microscopy: Phase, Nomarski
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From: taylor_cavanah-at-dcgsystems.com
Date: Thu, 24 Jun 2010 18:00:30 -0500
Subject: [Microscopy] viaWWW: Product Application Engineer

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

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: taylor_cavanah-at-dcgsystems.com
Name: Taylor Cavanah

Organization: DCG Systems - nanoInstruments Division

Title-Subject: [Filtered] Product Application Engineer

Message: There is an open position for a Product Applications
Engineer to work in our nanoInstruments Division. The position
involves heavy use of our SEM based Nanoprobing system. A microscopy
background is highly valuable as well as an electrical
characterization background. Full details can be found through this
link:
http://www.dcgsystems.com/positions_texas_Product%20Application%20Engineer.022210.html

Thank you for your time and any recommendations are appreciated.

Taylor Cavanah
General Manager, nI Division
469 226 2494

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8, 11 -- Subject: viaWWW: Product Application Engineer
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From: dac-at-research.umass.edu
Date: Fri, 25 Jun 2010 08:44:41 -0500
Subject: [Microscopy] Re: viaWWW: Optical Microscopy: Phase, Nomarski - more

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

Becca,

I can't say enough about the excellent resources for microscopy methods
available for self-study on the Molecular Expressions website, the Nikon
MicroscopyU site, and the nice documents that have been released by
Olympus and Zeiss. There are good explanations, graphics and Java
animations (some interactive).

One link for the Molecular Expressions is the "Primer" page
http://micro.magnet.fsu.edu/primer/
but click on the Special Techiques and other buttons at the left of that
page; that's where you will find the DIC and Phase Contrast material.
ALL of the material is first class. Much of this material is echoed on
the Nikon website under license and includes info on their microscope
systems as well. http://www.microscopyu.com/

Olympus has the publication "Microscope Basics and Beyond.pdf
(MortimerAbramowitz). This is 20Mb and is probably found on the Olympus
website. I can send a copy if you can't locate it.

And Zeiss has the publication "Microscopy - From The Very
Beginning.pdf", again, probably found on their website.

A good course can be helpful to some, but these materials are an
unbelievable resource and all for free.

Hope this helps,

Dale Callaham
Umass-at-Amherst

Email: rstiger-at-ppg.com
} Name: Rebecca Stiger
}
} Organization: PPG
}
} Title-Subject: [Filtered] Optical Microscopy: Phase, Nomarski
}
} Message: We're in a situation where we could really use some training
} in theory and practice of Nomarski and Phase Contrast optical
} imaging. I've been in touch with McCrone, but they seem to focus on
} Polarized Light Microscopy and it's applications.
}
} Any suggestions on good training for Nomarski and/or Phase? It would
} be nice to get all three techniques bundled into one event, but I
} don't know if it's possible.
}
} Many thanks,
}
} Becca
}


==============================Original Headers==============================
10, 18 -- From dac-at-research.umass.edu Thu Jun 24 20:11:20 2010
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10, 18 -- From: Dale Callaham {dac-at-research.umass.edu}
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From bridge-at-cools.com Thu Jun 24 22:57:28 2010
Return-Path: {bridge-at-cools.com}
Received: from host-70-45-82-92.onelinkpr.net (host-70-45-82-92.onelinkpr.net [70.45.82.92])
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Thu, 24 Jun 2010 22:57:26 -0500
Received: from [70.45.82.92] by cools.com; Thu, 24 Jun 2010 23:26:51 -0430

Becca,

I forgot to mention that DIC depends totally on polarization and it
could be that the McCrone Inst course deals with it that way. The
resources I mentioned below are excellent to generally understand the
workings of DIC and Phase Contrast. For a more complete and quantitative
approach, the books by Elizabeth Slayter are excellent. It is possible
to find new and used copies on Amazon, the used ones are priced very
reasonably. I think that the earlier (1976) edition of the Slayter book
covers all you need and there was recently a copy for $18.

Dale


I can't say enough about the excellent resources for microscopy methods
available for self-study on the Molecular Expressions website, the Nikon
MicroscopyU site, and the nice documents that have been released by
Olympus and Zeiss. There are good explanations, graphics and Java
animations (some interactive).

One link for the Molecular Expressions is the "Primer" page
http://micro.magnet.fsu.edu/primer/
but click on the Special Techiques and other buttons at the left of that
page; that's where you will find the DIC and Phase Contrast material.
ALL of the material is first class. Much of this material is echoed on
the Nikon website under license and includes info on their microscope
systems as well. http://www.microscopyu.com/

Olympus has the publication "Microscope Basics and Beyond.pdf
(MortimerAbramowitz). This is 20Mb and is probably found on the Olympus
website. I can send a copy if you can't locate it.

And Zeiss has the publication "Microscopy - From The Very
Beginning.pdf", again, probably found on their website.

A good course can be helpful to some, but these materials are an
unbelievable resource and all for free.

Hope this helps,

Dale Callaham
Umass-at-Amherst

Email: rstiger-at-ppg.com
} Name: Rebecca Stiger
}
} Organization: PPG
}
} Title-Subject: [Filtered] Optical Microscopy: Phase, Nomarski
}
} Message: We're in a situation where we could really use some training
} in theory and practice of Nomarski and Phase Contrast optical
} imaging. I've been in touch with McCrone, but they seem to focus on
} Polarized Light Microscopy and it's applications.
}
} Any suggestions on good training for Nomarski and/or Phase? It would
} be nice to get all three techniques bundled into one event, but I
} don't know if it's possible.
}
} Many thanks,
}
} Becca
}


==============================Original Headers==============================
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From: kenconverse-at-qualityimages.biz
Date: Fri, 25 Jun 2010 09:22:57 -0500
Subject: [Microscopy] viaWWW: Service on JSM 6100

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

Rod,
Any hints as to where you're located? It sounds like you want physical help
with the installation, so location is important.

How do you know you got to 8 mT? What gauging were you using? The JEOL
gauge will show 8 µA, which is different. If the turbo was running, 8 mT is
a pretty poor showing.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


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Email: vwporsche-at-verizon.net
Name: Rod Rowland

Organization: MATSYS, Inc.

Title-Subject: [Filtered] Service on JSM 6100

Message: I have a JEOL JSM 6100 that I bought on auction.
So far I have rebuilt the original mechanical pump.
Removed, checked, and reinstalled the turbo pump. Pulled
a vacuum to 8 millitorr. Now I'm looking for someone who
can help finish the installation. Please contact me via email.
Thanks
Rod

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From: ALawrence-at-entomology.msstate.edu
Date: Mon, 28 Jun 2010 05:33:31 -0500
Subject: [Microscopy] Microscopy and Microanalysis meeting in Portland

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

Hi, Becca

I wrote an article in American Lab way back in 1988 which explains (a) how DIC works (b) how to set up it and (c) how to avoid artifacts and mis-interpreting the results. You can find a PDF of a copy (my apologies.. a bit yellow with age) on our website: www.MicroscopyEducation.com.

As for phase... I can send you some of the teaching materials I've used for years, if that would be helpful.

Best regards,
Barbara Foster, President and Sr. Consultant

Microscopy/Microscopy Education
7101 Royal Glen Trail, Suite A
McKinney TX 75070
P: (972)924-5310 Skype: fostermme
W: www.MicroscopyEducation.com

NEWS! Visit the NEW and IMPROVED www.MicroscopyEducation.com! And don't forget: MME is now scheduling customized, on-site courses for the Summer and Fall. Call me for a free assessment and quote.

At 09:00 AM 6/25/2010, dac-at-research.umass.edu wrote:



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From nflodge-at-nccray.com Fri Jun 25 13:06:31 2010
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Email: stephen-at-Protochips.com
Name: Stephen Mick

Organization: Protochips, Inc

Title-Subject: [Filtered] Job Opening

Message: Protochips, a manufacturer and distributor of accessory
systems for environmental in situ Electron Microscopy is hiring an
additional Field Applications Specialist to support our product sales
efforts. This will be a technical position with significant customer
interaction working with researchers desiring to bring a working
environment (heat, electrical, liquid and gas), into their real-time
EM research. For more information please go to
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From kefwyomingbankdiz-at-wyomingbank.com Sat Jun 26 04:55:17 2010
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Kristen:

Simple crystals mimic their atomic arrangement by their shape or surface
texture. Salt and sugar are easy to come by. Some sand can be
interesting. Mineral samples also often have interesting shapes that
mimic their crystal structure.

Velcro is a also fun, especially if you can find a sand burr for
comparison. This shows a functional microscopic shape and how man can
mimic nature.

I have found that most high school teachers don't get that excited about
the fractures and microstructures that I think are cool. That said,
polished metal specimens with distinct phases with greatly different
density can be great for demonstrating backscattered electron images.
Brass with lead inclusions, copper pipe with a solder joint, cross
sections of circuit boards, all have high contrast BSE images. Maybe one
your colleagues in the engineering dept can find something laying around
for you.

Good luck and have fun.

--
Larry D. Hanke, P.E.
Materials Evaluation and Engineering, Inc.
Practical Solutions Through Technology and Innovation
http://www.mee-inc.com (763) 449-8870

} Hi listers,
}
} In a turn of events that will make many of you smile knowingly, I've been asked to give a half day, hands-on "workshop" on "EM" as a professional development activity for some local high school science teachers. It's a great idea, but, as you all well know, there is just no such thing as throwing something on the SEM or TEM for a "quick pic"! Thankfully, we have an SEM with LV capabilities so that we can look at unfixed samples!
}
} Some of these teachers are physics teachers and so probably not so excited about looking at the biological samples with which I am well-acquainted and bringing the micrographs back to their students. Do any of the materials scientists out there have a suggestion for an easy (and easily available) materials sample to look at with these teachers?
}
} Thanks in advance for any advice that you can offer,
} Kristen Lennon
} Frostburg State University
} Dept of Biological Sciences
} Frostburg, MD 21532
} kalennon-at-frostburg.edu
} http://www.frostburg.edu/dept/biol/


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The 2010 Microscopy and Microanalysis meeting is 1 month away and we
need your help to make sure things run smoothly. Student bursaries or
volunteers are needed to do things such as provide support in the
different symposia (helping with audio-visual needs, maintaining an
attendance count, and helping speakers set up for their presentation),
staff the MSA MegaBooth or volunteer office, monitor use of the Internet
Café, and help with vendor tutorials

Students will be paid $10/hour for assisting with any of the tasks
mentioned above (paid by check at the end of the meetings). There is an
added bonus of $10 cash for each morning and/or afternoon session worked
to help with meals. Volunteers will also receive some compensation to
help with meeting expenses and are eligible for the $10 cash for meals.

We can*t do it without help from you, so please consider serving as a
student bursary or volunteer. If anyone has any questions about the
bursary/volunteer program, or would like to participate, please
contact:

Amanda Lawrence
Electron Microscope Center
Mississippi State University
662-325-3019
alawrence-at-entomology.msstate.edu




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From: cpawlowicz-at-ubmtechinsights.com
Date: Mon, 28 Jun 2010 13:04:52 -0500
Subject: [Microscopy] TEM JEOL 2011 STEM/EDS

Contents Retrieved from Microscopy Listserver Archives
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Email: heller-at-uni.hohenheim.de
Name: Heller, Anne

Organization: Universit”t Hohenheim

Title-Subject: [Filtered] TEM Preparation of chromoplasts

Message: Has anybody experience in the
preparation of chromoplasts for transmission
electron microscopy?

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From ngpaulson-at-hotmail.com Mon Jun 28 07:58:04 2010
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Hi,

We have a JEOL 2011 TEM and are looking to add STEM and EDS.

JEOL no longer offers this accessory so we're looking for alternatives- can anybody suggest third party vendors that might offer this? or does anybody have a STEM system sitting unused that they'd be interested in selling (with or without EDS)?

thanks!

Chris Pawlowicz, B.Eng
Manager, Lab Development
UBM TechInsights
3000 Solandt Rd Ottawa ON Canada K2K 2X2
T: +1 613 576 0150
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From: sgkcck-at-aol.com
Date: Mon, 28 Jun 2010 22:52:31 -0500
Subject: [Microscopy] viaWWW: Aurion ImmunoGold Silver Staining Workshop

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Hi listers,
I have heard about tabletop SEM developed in Oslo University Medical
School somewhere in nineteen sixties or seventies.
Any information about this instrument and its developers will be highly
appreciated.
Thanks,
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784


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From intake-at-dntxlegal.com Mon Jun 28 20:57:32 2010
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Email: sgkcck-at-aol.com
Name: Stacie Kirsch

Organization: Electron Microscopy Sciences

Title-Subject: [Filtered] Aurion ImmunoGold Silver Staining Workshop

Message: The Aurion Immuno Gold Silver Staining Workshop
Pre Microscopy & Microanalysis 2010 Workshop
July 28-30 , 2010

Aurion and Electron Microscopy Sciences will hold their workshop on
Immuno Gold Silver Staining at the House Ear Institute, Los Angeles
California from July 28-30, 2010 prior to Microscopy & Microanalysis
2010. The course objectives are to provide researchers with the
opportunity to learn the theory and practice of immunogold labeling
and to promote technology exchange and research collaboration by
allowing participants to process their own samples under expert
guidance.

The Immuno Gold Silver Staining Workshop is designed and taught by
Mr. Peter van de Plas who has been with Aurion since 1991. Mr. van de
Plas worked closely together with Dr. Leunissen in founding a firm
basis for Aurion including development of product applications. He
has been invited to many international microscopy conferences and
workshops and is especially experienced in providing hands-on
training. Dr. Paul Webster, head of Ahmanson Advanced EM & Imaging
Center at the at the House Ear Institute, will be providing expertise
in immunolabeling and applications of electron microscopy to
biomedical research.

The three day course will cover the properties of gold particles and
their protein conjugates, theories underlying immunogold labeling
protocols, and silver enhancement of gold particles. Also covered in
the course will be immunogold labeling on a variety of sample
preparations for LM, immunogold labeling for EM, pre/post-embedding
double immunogold labeling, and background minimization in immunogold
labeling.
Participants are encouraged to bring their own samples to maximize
learning. Please visit the course site for additional details:
http://www.emsdiasum.com/microscopy/products/immunogold/workshop.aspx

For more information contact:
Stacie Kirsch
stacie-at-ems-secure.com
215-412-8402
www.emsdiasum.com/microscopy


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11, 11 -- To: microscopy-at-microscopy.com
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11, 11 -- Subject: viaWWW: Aurion ImmunoGold Silver Staining Workshop
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From: cgsellers-at-temple.edu
Date: Mon, 28 Jun 2010 22:52:54 -0500
Subject: [Microscopy] viaWWW: TEM preparation of Dinoflagellates, Haptophyte algae

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Email: cgsellers-at-temple.edu
Name: C. Grier Sellers

Organization: Dept. of Biology, Temple University

Title-Subject: [Filtered] TEM preparation of Dinoflagellates, Haptophyte algae

Message: I wonder if anyone has any experience with preparation of
dinoflagellates, in particular, unarmored marine dinoflagellates for
TEM.

Also, if you have any experience with marine Haptophyte (Prymnesiophyte) algae.

Thanks,

Grier

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9, 11 -- Subject: viaWWW: TEM preparation of Dinoflagellates, Haptophyte algae
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From: kbaldwin-at-howard.edu
Date: Mon, 28 Jun 2010 22:53:38 -0500
Subject: [Microscopy] viaWWW: surplus osmium

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Email: kbaldwin-at-howard.edu
Name: Dr. Kate Baldwin

Organization: Howard University

Title-Subject: [Filtered] surplus osmium

Message: I am a former longñtime member of CSM
and am about to retire from Howard University.
Over the years I have acquired (inherited,
bought) a lot of osmium in 1 g sealed ampoules
(15 g total).

I would like to find someone in the Washington DC
area who could use it rather than dispose of it
in the hazardous waste. According to a post by
Tom Phillips in the MSA newsletter, he has used
even older osmium and it was fine.

I also have a couple liters of old Epon 812. My
recollection is that this also has a very long
shelf life and should be good as well.

I work in DC, but live in Bethesda so could
easily deliver it to pretty much anyone in the
area. I would be grateful if anyone who knows of
a lab in the DC area that could use these
chemicals would let me know.

Kate Baldwin


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From: tina-at-pbrc.hawaii.edu
Date: Tue, 29 Jun 2010 14:20:57 -0500
Subject: [Microscopy] 3D modeling from SEM

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Email: ahmad_ds-at-yahoo.com
Name: Ahmad Ashkaibi

Organization: Al-Balqa Applied University

Title-Subject: [Filtered] RE: table top SEM

Message: Hi Vladimir,

You can get a lot of information about table top SEM at the following website:

http://www.fei.com/products/scanning-electron-microscopes/phenom.aspx

you can also watch a movie talking about it.

Regards,

Ahmad Ashkaibi

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From jacksom3-at-ocps.k12.fl.us Tue Jun 29 02:57:15 2010
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Hi Debby:

If you 'insist' to use yeast, I have yeast cells embedded in resin (Embed 812) and I'd be happy to send you a couple of blocks. You can cut 2-3 um sections and the sections would stay easily for several years, if not more.

Let me know if you are interested.

Zhaojie


Zhaojie Zhang, Ph. D.
Director, Microscopy Core Facility
University of Wyoming
Laramie, WY 82071
PHONE: 307-766-3038
FAX: 307-766-5625



-----Original Message-----
X-from: dsherman-at-purdue.edu [mailto:dsherman-at-purdue.edu]
Sent: Sunday, June 20, 2010 8:39 PM
To: Z.J. Zhang

Hi all,

Our beginning Biology course has a lab that requires slides for counting
cells using a micrometer eyepiece on a light microscope. They want to use
yeast but found that preparing fresh samples during the lab took too much
time and was so variable that the students did not get to complete the lab
on time. Thus they would like to make permanent slides that can be used in
the labs for a number of years.

Suggestions as to how to prepare slides (about 50 of them) containing a
monolayer of yeast stained with hemotoxylin would be appreciated.

Debby


---
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy




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9, 28 -- To: "message to: MSA list" {microscopy-at-microscopy.com}
9, 28 -- Date: Sun, 20 Jun 2010 22:33:59 -0400
9, 28 -- Subject: Yeast slides for LM
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23, 29 -- From ZZhang-at-uwyo.edu Tue Jun 29 09:26:59 2010
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From kefxanthdiz-at-xanth.net Tue Jun 29 13:40:41 2010
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Hi, All-

It seems to me this has been discussed in the fairly recent past, but I
can't find the posts. Two mechanical engineering students (whose major
professors have disappeared for the summer) have been tasked with making a
3D model of a larval copepod with images taken in the SEM. I was told they
had appropriate software. In fact, after I trained them on the SEM and
then asked them how many degrees between shots and at what angles they
needed (and I know darn well I should have insisted on doing this first),
it turns out the two software packages they had been given are, according
to the software manufacturers, not appropriate.

The best we can figure after Googling around is that this might be
possible somewhere down the line, with each image being made up of stereo
pairs to help compensate for "unusual optical properties", by which I
suspect they mean foreshortening. I haven't investigated any further,
since troubleshooting this is what the students are supposed to do all
summer, but I'm still inclined to help.

Any ideas?

Oh, and after the modeling, they are supposed to figure out what happens
as this organism swims through water... If you've ever seen these hairy
little beasts, you'll know this is a bit crazy. So far what I've done is
have them take images every 45 degrees around a small hex nut, tilted so
that is also shows the inside threads. The stereo pairs look cool. I told
them to model what happens as the nut moves through water, especially as
the water goes through the threaded hole. I told them to look for the ad
on TV about the spiral neck on some beer bottle that affects the flow of
the beer out of the bottle to enhance your enjoyment...

Aloha,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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From: holpc-at-firstenergycorp.com
Date: Tue, 29 Jun 2010 14:38:46 -0500
Subject: [Microscopy] Cathodic cleaning of oxidized surfaces

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


I am working on cleaning some heavily oxidized fracture surface(s) and have
been asked to try and clean cathodically since neither alkaline solutions
nor inhibited acid has worked satisfactorily.
I'm using a sodium carbonate solution right now with marginal success, so I
was wondering if anyone had any favorite or recommended solutions. I would
appreciate any and all suggestions.

Thanks,

Chris Holp
FirstEnergy Corp.
BETA Labs

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From: bozzola-at-siu.edu
Date: Tue, 29 Jun 2010 15:14:20 -0500
Subject: [Microscopy] EM: Life Expectancy of an EM Unit

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

I had a lively discussion recently with an administrator regarding the
useful life expectancy of an electron beam instrument.

It would be great of such instruments lasted a really long time, but
they don't.

Here are some questions:

1. What has been your personal experience with useable lifetimes of
electron microscopes (SEM and/or TEM)? Feel free to rant and rave
(especially if you email me directly).

2. Do newer instruments become obsolete at a faster rate than did the
older instruments? Observations and comments.

To preserve the good will of our sponsors, please email me directly
(if brand names are given) and I will publish a generic version here.

Thank you.



--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

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From: holpc-at-firstenergycorp.com
Date: Tue, 29 Jun 2010 15:53:06 -0500
Subject: [Microscopy] Cathodic cleaning of oxidized surfaces: base material

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


My apologies to the group.

The base material for this oxidized fracture is a carbon steel (0.27% C).
There are some weldments involved with similar composition.

Chris

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From: kenconverse-at-qualityimages.biz
Date: Tue, 29 Jun 2010 17:11:23 -0500
Subject: [Microscopy] Cathodic cleaning of oxidized surfaces: base material

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

Chris,
Have you tried 70% phosphoric acid? It'll remove the oxide without touching
the base metal. Alternatively, you can use Coca Cola and lose a little of
the base metal. It's not quite 70%.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: holpc-at-firstenergycorp.com [mailto:holpc-at-firstenergycorp.com]
Sent: Tuesday, June 29, 2010 4:55 PM
To: kenconverse-at-qualityimages.biz


My apologies to the group.

The base material for this oxidized fracture is a carbon steel (0.27% C).
There are some weldments involved with similar composition.

Chris

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From: kenconverse-at-qualityimages.biz
Date: Tue, 29 Jun 2010 17:28:39 -0500
Subject: [Microscopy] EM: Life Expectancy of an EM Unit

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

Hi John,
My experience has been that 30-40 years is not unreasonable for some of the
ETECs, Amrays and JEOLs that were sold in the 70s (and maybe Hitachis and
some others).

30-40 years is probably not unreasonable for the "hardware" side of the
newer systems, but can you keep a computer running it for that amount of
time? I doubt it.

Given that a PC, when installed, is already obsolete, you're down to about a
5 year life span, what with how operating systems get changed almost as
often as your socks. And the OS companies are fighting to shorten their
support cycle for their previous disasters.

Now, consider that SEM manufacturers are not writing software for the latest
OS, but probably 1 or 2 generations back, and they may, or may not, write
software for a newer OS. If they do, you're going to pay about $25k for a
$600 desktop computer and some new software for their SEM.

No one, and I mean no one, is going to be talking about their SEM that they
bought new in 2010 and is still doing yeoman's service in 2030, let alone
2050. On the other hand, there may still be a few '70s vintage scopes still
running then, because they are not dependent on a PC (and they can also be
serviced by third party service companies). You and Paul Robinson got rid
of your ETEC Autoscan way too early. Such is life.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: bozzola-at-siu.edu [mailto:bozzola-at-siu.edu]
Sent: Tuesday, June 29, 2010 4:16 PM
To: kenconverse-at-qualityimages.biz

I had a lively discussion recently with an administrator regarding the
useful life expectancy of an electron beam instrument.

It would be great of such instruments lasted a really long time, but
they don't.

Here are some questions:

1. What has been your personal experience with useable lifetimes of
electron microscopes (SEM and/or TEM)? Feel free to rant and rave
(especially if you email me directly).

2. Do newer instruments become obsolete at a faster rate than did the
older instruments? Observations and comments.

To preserve the good will of our sponsors, please email me directly
(if brand names are given) and I will publish a generic version here.

Thank you.



--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

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From: mlibbee-at-gmail.com
Date: Tue, 29 Jun 2010 20:08:49 -0500
Subject: [Microscopy] viaWWW: AuPd v. Pt grain size

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Email: mlibbee-at-gmail.com
Name: Marissa Libbee

Title-Subject: [Filtered] AuPd v. Pt grain size

Message: In regards to sputter coating... will someone please inform
me if anyone has ever compared the grain size of Pt to AuPd,
particularly in terms of HRES SEM analysis?

Thanks!

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From: tylerjhayden-at-gmail.com
Date: Tue, 29 Jun 2010 20:09:18 -0500
Subject: [Microscopy] rviaWWW: Microfibril preparation for SEM/ESEM

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Email: tylerjhayden-at-gmail.com
Name: Tyler Hayden

Organization: University Lincoln-Nebraska

Title-Subject: [Filtered] Microfibril preparation for SEM/ESEM

Message: I'm looking for a method to prepare a leaf sample for the
SEM to visualize the microfibril orientation. Any ideas or past
experience on the subject?

Tyler

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From: r-holdford-at-ti.com
Date: Tue, 29 Jun 2010 22:40:14 -0500
Subject: [Microscopy] Re: viaWWW: AuPd v. Pt grain size

Contents Retrieved from Microscopy Listserver Archives
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Marissa: I presented on this very topic at SCANNING in 2006. The Pt
grain size is almost undetectable at 2kV, 300KX magnification in a
Hitachi S-5200 FE-SEM. AuPd (as is Au) is very noticeable under these
conditions. Pt, Cr, and Ir all have about the same surface quality. In
my notes I have the approximate grain size of AuPd to be around 1.8nm
and Pt to be around 1.5nm using the conditions in my Emitech K575X
sputter coater. Sputter coating variables will have an effect on the
grain sizes of the metals.

On 6/29/10 8:09 PM, mlibbee-at-gmail.com wrote:
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} Email: mlibbee-at-gmail.com
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} Title-Subject: [Filtered] AuPd v. Pt grain size
}
} Message: In regards to sputter coating... will someone please inform
} me if anyone has ever compared the grain size of Pt to AuPd,
} particularly in terms of HRES SEM analysis?
}
} Thanks!
}
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--
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Becky Holdford (r-holdford-at-ti.com)
214-567-0360
SC Packaging FA
Texas Instruments, Inc.
13536 N. Central Expressway MS 940
Dallas, TX 75243
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From: gas19-at-chrysler.com
Date: 06/29/2010 03:28PM
Subject: [Microscopy] 3D modeling from SEM

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

A seemingly simple question but a not simple answer.

Au coating is like Spiderman. Au/Pd is good but not
as good at Pt, Pd or Ir. But the big variable is the
terminal vacuum. Poor vacuum (30mT) is not going to be
able to put down an invisible coating that would or could
be done at 15mT. That is just vacuum. Longer coating
time at lower current will improve the coating.

At 200KX-300KX mag on a Zeiss FESEM I can see the Au/Pd
coating. Bad. Not so with Pd or Ir at 15mT. But I do find
that it takes a longer coating to achieve a good result.
This is using a Denton Desk IV TSC turbo coater with
tilt and rotate.

In conclusion, I would prefer the Pt over Au/Pd but search
for Pd or Ir in its replacement. Recovery in Las Vegas
has all of these targets. Good prices.

gary g.


At 06:11 PM 6/29/2010, you wrote:
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8, 20 -- From gary-at-gaugler.com Tue Jun 29 23:25:13 2010
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From gguson-at-tgedu.net Tue Jun 29 23:59:50 2010
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Hi Ken

I 100% agree with you, that it is the dreaded computer that has not only
caused a very rapid advance in electron microscope performance but also has
caused the death of so many!

As a standard procedure I train service engineers that when installing a new
instrument they should make everyone conversant with problems in the future.
We point out that during the life of the new microscope the staff will
surely change their desk top computers several times and that by the time
the microscope computer starts to give problems it may be almost impossible
to source the parts required. The solution is not to throw away the E.M.
unit computers being used when the installation took place, but save the
CPU, keyboard and even the mouse, as one day they may well be a very
valuable source of spare parts as your instrument ages.

A simple but only partial solution to a problem that becomes even bigger
with each new generation of computers!

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com

-----Original Message-----
X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz]
Sent: 29 June 2010 23:29
To: protrain-at-emcourses.com

Hi John,
My experience has been that 30-40 years is not unreasonable for some of the
ETECs, Amrays and JEOLs that were sold in the 70s (and maybe Hitachis and
some others).

30-40 years is probably not unreasonable for the "hardware" side of the
newer systems, but can you keep a computer running it for that amount of
time? I doubt it.

Given that a PC, when installed, is already obsolete, you're down to about a
5 year life span, what with how operating systems get changed almost as
often as your socks. And the OS companies are fighting to shorten their
support cycle for their previous disasters.

Now, consider that SEM manufacturers are not writing software for the latest
OS, but probably 1 or 2 generations back, and they may, or may not, write
software for a newer OS. If they do, you're going to pay about $25k for a
$600 desktop computer and some new software for their SEM.

No one, and I mean no one, is going to be talking about their SEM that they
bought new in 2010 and is still doing yeoman's service in 2030, let alone
2050. On the other hand, there may still be a few '70s vintage scopes still
running then, because they are not dependent on a PC (and they can also be
serviced by third party service companies). You and Paul Robinson got rid
of your ETEC Autoscan way too early. Such is life.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: bozzola-at-siu.edu [mailto:bozzola-at-siu.edu]
Sent: Tuesday, June 29, 2010 4:16 PM
To: kenconverse-at-qualityimages.biz

I had a lively discussion recently with an administrator regarding the
useful life expectancy of an electron beam instrument.

It would be great of such instruments lasted a really long time, but
they don't.

Here are some questions:

1. What has been your personal experience with useable lifetimes of
electron microscopes (SEM and/or TEM)? Feel free to rant and rave
(especially if you email me directly).

2. Do newer instruments become obsolete at a faster rate than did the
older instruments? Observations and comments.

To preserve the good will of our sponsors, please email me directly
(if brand names are given) and I will publish a generic version here.

Thank you.



--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

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From mitzi.mclaurine-at-brewtech.org Wed Jun 30 06:47:45 2010
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I have seen a demonstration of how a 3D reconstruction can be made using the results from a 4 quadrant BSE detector. Somehow the computer can take the results from each of the quadrants of the detector and make a 3D model where measurements can be made for depth and angle. I think I remember it was from Hitachi, so you might check their website for an online demo. An example was a screw where measurement of the threads could be made. Pretty cool, just too expensive to consider.

Maybe something like this could help them with their 3D model of the critter.

Gerry Shulke
Materials Engineering Specialist
Materials Characterization Labs
Chrysler Group LLC

-----tina-at-pbrc.hawaii.edu wrote: -----


To: gas19-at-chrysler.com
X-from: tina-at-pbrc.hawaii.edu




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Hi, All-

It seems to me this has been discussed in the fairly recent past, but I
can't find the posts. Two mechanical engineering students (whose major
professors have disappeared for the summer) have been tasked with making a
3D model of a larval copepod with images taken in the SEM. I was told they
had appropriate software. In fact, after I trained them on the SEM and
then asked them how many degrees between shots and at what angles they
needed (and I know darn well I should have insisted on doing this first),
it turns out the two software packages they had been given are, according
to the software manufacturers, not appropriate.

The best we can figure after Googling around is that this might be
possible somewhere down the line, with each image being made up of stereo
pairs to help compensate for "unusual optical properties", by which I
suspect they mean foreshortening. I haven't investigated any further,
since troubleshooting this is what the students are supposed to do all
summer, but I'm still inclined to help.

Any ideas?

Oh, and after the modeling, they are supposed to figure out what happens
as this organism swims through water... If you've ever seen these hairy
little beasts, you'll know this is a bit crazy. So far what I've done is
have them take images every 45 degrees around a small hex nut, tilted so
that is also shows the inside threads. The stereo pairs look cool. I told
them to model what happens as the nut moves through water, especially as
the water goes through the threaded hole. I told them to look for the ad
on TV about the spiral neck on some beer bottle that affects the flow of
the beer out of the bottle to enhance your enjoyment...

Aloha,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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From: DennisH658-at-aol.com
Date: Wed, 30 Jun 2010 08:51:35 -0500
Subject: [Microscopy] Re: 3D modeling from SEM

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Wed, 30 Jun 2010 08:51:35 -0500

Tina:

If you're referring to 3-D, involving 2 pictures and (normally) a special
pair of glasses, StereoPhoto Maker is highly recommended.
(stereo.jpn.org/eng/stphmkr/). The two pictures need photographed at approximately 16
degrees apart, though a slightly greater angle will do. (Increasing the angle
gives the picture more depth.)

The two pictures can be "freeviewed," if they're approximately 65mm from
center-to-center (the distance between the pupils of the eyes) and each eye
focuses straight ahead at the appropriate picture. Or switch the pictures
around and you can look at them cross-eyed.

Dennis
=========================
In a message dated 6/30/2010 8:43:28 A.M. Eastern Daylight Time,
gas19-at-chrysler.com writes:
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Hi, All-

It seems to me this has been discussed in the fairly recent past, but I
can't find the posts. Two mechanical engineering students (whose major
professors have disappeared for the summer) have been tasked with making a
3D model of a larval copepod with images taken in the SEM. I was told they
had appropriate software. In fact, after I trained them on the SEM and
then asked them how many degrees between shots and at what angles they
needed (and I know darn well I should have insisted on doing this first),
it turns out the two software packages they had been given are, according
to the software manufacturers, not appropriate.

The best we can figure after Googling around is that this might be
possible somewhere down the line, with each image being made up of stereo
pairs to help compensate for "unusual optical properties", by which I
suspect they mean foreshortening. I haven't investigated any further,
since troubleshooting this is what the students are supposed to do all
summer, but I'm still inclined to help.

Any ideas?

Oh, and after the modeling, they are supposed to figure out what happens
as this organism swims through water... If you've ever seen these hairy
little beasts, you'll know this is a bit crazy. So far what I've done is
have them take images every 45 degrees around a small hex nut, tilted so
that is also shows the inside threads. The stereo pairs look cool. I told
them to model what happens as the nut moves through water, especially as
the water goes through the threaded hole. I told them to look for the ad
on TV about the spiral neck on some beer bottle that affects the flow of
the beer out of the bottle to enhance your enjoyment...

Aloha,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu
*
* Biological Electron Microscope Facility * (808) 956-6251
*
* University of Hawaii at Manoa *
http://www.pbrc.hawaii.edu/bemf*
****************************************************************************



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From: b-myers3-at-northwestern.edu
Date: Wed, 30 Jun 2010 09:21:04 -0500
Subject: [Microscopy] Re: 3D modeling from SEM

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

Hi Tina-

You should check out the Mex software from Alicona.

http://www.alicona.com/englisch/main-navigation/products/mex/index.html

We bought this software (which is not particularly cheap) and it does a
reasonably good job of rendering 3-D surfaces from stereo pairs. It has
a wide range of measurement tools for various types of analysis.
However, as you might imagine it does not do a very good job with high
aspect ratio or complex 3-D structures.

You might also check out Scandium Solution Height or 3D_TOPx, although
I've used neither of these.

Regards,
Ben



On 6/29/2010 2:31 PM, tina-at-pbrc.hawaii.edu wrote:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi, All-
}
} It seems to me this has been discussed in the fairly recent past, but I
} can't find the posts. Two mechanical engineering students (whose major
} professors have disappeared for the summer) have been tasked with making a
} 3D model of a larval copepod with images taken in the SEM. I was told they
} had appropriate software. In fact, after I trained them on the SEM and
} then asked them how many degrees between shots and at what angles they
} needed (and I know darn well I should have insisted on doing this first),
} it turns out the two software packages they had been given are, according
} to the software manufacturers, not appropriate.
}
} The best we can figure after Googling around is that this might be
} possible somewhere down the line, with each image being made up of stereo
} pairs to help compensate for "unusual optical properties", by which I
} suspect they mean foreshortening. I haven't investigated any further,
} since troubleshooting this is what the students are supposed to do all
} summer, but I'm still inclined to help.
}
} Any ideas?
}
} Oh, and after the modeling, they are supposed to figure out what happens
} as this organism swims through water... If you've ever seen these hairy
} little beasts, you'll know this is a bit crazy. So far what I've done is
} have them take images every 45 degrees around a small hex nut, tilted so
} that is also shows the inside threads. The stereo pairs look cool. I told
} them to model what happens as the nut moves through water, especially as
} the water goes through the threaded hole. I told them to look for the ad
} on TV about the spiral neck on some beer bottle that affects the flow of
} the beer out of the bottle to enhance your enjoyment...
}
} Aloha,
} Tina
}
} ****************************************************************************
} * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
} * Biological Electron Microscope Facility * (808) 956-6251 *
} * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
} ****************************************************************************
}
} ==============================Original Headers==============================
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--
*Ben Myers*
SEM/FIB Facility Manager
NU/ANCE/ Center

Northwestern University
Mail: 2036 Cook Hall
Office: 1114 Cook Hall
2220 Campus Drive
Evanston, IL 60208-3108

ph: (847) 491-3439
fax: (847) 467-6573

http://www.nuance.northwestern.edu

==============================Original Headers==============================
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From: TindallR-at-missouri.edu
Date: Wed, 30 Jun 2010 09:32:14 -0500
Subject: [Microscopy] EM: Life Expectancy of an EM Unit

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

Dr. B,

I wholeheartedly agree with those who say that the limitation is the computer and software. We have one instrument running on Windows NT Workstation 4.0. To upgrade it to an OS from this century would cost an estimated $30-40K (yes, you read that right).

On the other hand, you might still have the old Hitachi S570 running that I started on back in 1987. The Hitachi HU11's, H500, and probably the H7100 would have run as long as parts and service are available. And our JEOL 1200EX here was still going after 20+ years of hard use----getting creaky, but still running and producing good results.

I think one of the biggest problems the OEMs should be addressing now is making the software and computer components of their instruments more upgradeable and updatable. I suspect there are financial as well as technical aspects to this problem, as in more turnover, more profits, but I'll let the OEMs comment on that aspect.

Rando

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




-----Original Message-----
X-from: bozzola-at-siu.edu [mailto:bozzola-at-siu.edu]
Sent: Tuesday, June 29, 2010 3:15 PM
To: Tindall, Randy D.

I had a lively discussion recently with an administrator regarding the
useful life expectancy of an electron beam instrument.

It would be great of such instruments lasted a really long time, but
they don't.

Here are some questions:

1. What has been your personal experience with useable lifetimes of
electron microscopes (SEM and/or TEM)? Feel free to rant and rave
(especially if you email me directly).

2. Do newer instruments become obsolete at a faster rate than did the
older instruments? Observations and comments.

To preserve the good will of our sponsors, please email me directly
(if brand names are given) and I will publish a generic version here.

Thank you.



--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730

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From: kraftpiano-at-gmail.com
Date: Wed, 30 Jun 2010 09:59:08 -0500
Subject: [Microscopy] EM: Life Expectancy of an EM Unit

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

Re: the "dreaded computer" problem of obsolescence of new machines -
is anyone working on the hardware integration/ engineering problems
required to get around this? It sounds very much like the kind of thing
a engineering department somewhere would probably be doing, collaborating
with manufacturing type people.

Surely, instead of having the computing architecture completely integrated
into the EM in many different and complex ways, it would actually be easy
to have just a few custom parts specific to the microscope, that were
integrated with the computer in very standard ways that are not likely to
go out of date soon. These custom parts driving the specific microscope
engineering could be connected to a bog-standard computer running some
nice stable (open-source) OS like Linux, that could be updated easily.
Then all that has to be updated with a new OS or computer is the
integration of the custom parts. This shouldn't be too expensive to get
right, given the cost of producing an EM anyway.

For connecting custom bits of computing/engineering to the actual computer
- by standard ways not likely to go out of date, I mean using firewire
connections instead of custom data cables, and obeying IEEE standards for
everything, say. Make it possible for the computing to be open-source,
and you'll get your product development done for free by other people...
probably faster...

Built-in 5-year obsolescence might be a good business model for computers,
but it must be prohibitively expensive for a lot of university departments
deciding whether to replace their old workhorse EMs. Surely the EM
companies would do better - from a business point of view - to provide an
EM as outlined above, that people could be relatively sure would keep
working for 10-20 years, and then lock users into expensive service
contracts because of the custom software/ drivers of the engineering,
needed with every new OS update for the computer? You sell
proportionately less to each customer, but I be the number of customers
more than makes up for it....

(Isn't this what happens with LMs anyway these days?)


Giselle


On Wed, 30 Jun 2010, protrain-at-emcourses.com wrote:

}
}
}
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} Hi Ken
}
} I 100% agree with you, that it is the dreaded computer that has not only
} caused a very rapid advance in electron microscope performance but also has
} caused the death of so many!
}
} As a standard procedure I train service engineers that when installing a new
} instrument they should make everyone conversant with problems in the future.
} We point out that during the life of the new microscope the staff will
} surely change their desk top computers several times and that by the time
} the microscope computer starts to give problems it may be almost impossible
} to source the parts required. The solution is not to throw away the E.M.
} unit computers being used when the installation took place, but save the
} CPU, keyboard and even the mouse, as one day they may well be a very
} valuable source of spare parts as your instrument ages.
}
} A simple but only partial solution to a problem that becomes even bigger
} with each new generation of computers!
}
} Steve Chapman FRMS
} Senior Consultant Protrain
} For consultancy and training in electron microscopy world wide
} Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
} www.emcourses.com
}
} -----Original Message-----
} X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz]
} Sent: 29 June 2010 23:29
} To: protrain-at-emcourses.com
} Subject: [Microscopy] RE: EM: Life Expectancy of an EM Unit
}
}
}
}
} ----------------------------------------------------------------------------
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}
} Hi John,
} My experience has been that 30-40 years is not unreasonable for some of the
} ETECs, Amrays and JEOLs that were sold in the 70s (and maybe Hitachis and
} some others).
}
} 30-40 years is probably not unreasonable for the "hardware" side of the
} newer systems, but can you keep a computer running it for that amount of
} time? I doubt it.
}
} Given that a PC, when installed, is already obsolete, you're down to about a
} 5 year life span, what with how operating systems get changed almost as
} often as your socks. And the OS companies are fighting to shorten their
} support cycle for their previous disasters.
}
} Now, consider that SEM manufacturers are not writing software for the latest
} OS, but probably 1 or 2 generations back, and they may, or may not, write
} software for a newer OS. If they do, you're going to pay about $25k for a
} $600 desktop computer and some new software for their SEM.
}
} No one, and I mean no one, is going to be talking about their SEM that they
} bought new in 2010 and is still doing yeoman's service in 2030, let alone
} 2050. On the other hand, there may still be a few '70s vintage scopes still
} running then, because they are not dependent on a PC (and they can also be
} serviced by third party service companies). You and Paul Robinson got rid
} of your ETEC Autoscan way too early. Such is life.
}
} Ken Converse
} owner
}
} QUALITY IMAGES
} Servicing Scanning Electron Microscopes
} Since 1981
} 474 So. Bridgton Rd.
} Bridgton, ME 04009
} 207-647-4348
} Fax 207-647-2688
} kenconverse-at-qualityimages.biz
} qualityimages.biz
}
}
} -----Original Message-----
} X-from: bozzola-at-siu.edu [mailto:bozzola-at-siu.edu]
} Sent: Tuesday, June 29, 2010 4:16 PM
} To: kenconverse-at-qualityimages.biz
} Subject: [Microscopy] EM: Life Expectancy of an EM Unit
}
}
}
}
} ----------------------------------------------------------------------------
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} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} I had a lively discussion recently with an administrator regarding the
} useful life expectancy of an electron beam instrument.
}
} It would be great of such instruments lasted a really long time, but
} they don't.
}
} Here are some questions:
}
} 1. What has been your personal experience with useable lifetimes of
} electron microscopes (SEM and/or TEM)? Feel free to rant and rave
} (especially if you email me directly).
}
} 2. Do newer instruments become obsolete at a faster rate than did the
} older instruments? Observations and comments.
}
} To preserve the good will of our sponsors, please email me directly
} (if brand names are given) and I will publish a generic version here.
}
} Thank you.
}
}
}
} --
} John J. Bozzola, Ph.D., Director
} I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
} Southern Illinois University
} 750 Communications Drive
} Carbondale, IL 62901
} Phone: 618-453-3730
}
} ==============================Original Headers==============================
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} {microscopy-at-microscopy.com}
} 26, 28 -- Subject: RE: [Microscopy] EM: Life Expectancy of an EM Unit
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} 37, 27 -- From protrain-at-emcourses.com Wed Jun 30 04:50:13 2010
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10, 26 -- From gw265-at-hermes.cam.ac.uk Wed Jun 30 09:47:35 2010
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From newsletter-at-deidox.com Wed Jun 30 09:51:37 2010
Return-Path: {newsletter-at-deidox.com}
Received: from [118.32.90.144] ([118.32.90.144])
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Received: from [118.32.90.144] by aspmx.l.google.com; Wed, 30 Jun 2010 23:50:45 +0900

I've been reading this thread with some interest, as in a previous
life, I was a computer consultant (Although I don't like admitting it
in public.) There are ways to make older software run on newer
operating systems and computers, and I wonder if the marriage between
the EM software and the computer is more of an artificial one.

For example, I received a JEOL DSG unit to use with the 35 that was
installed where I was teaching, and despite the software being written
for windows 3.1, it is now working perfectly on windows XP. The only
limitation that I've found with that system is that the hardware is an
old ISA board, but I'm sure that with the proper schematics (Which I
have) and time (Which I don't have) I could re-wire it for a more
modern PCI card and get similar results without too much hassle.

I don't see any reason why a modern EM, computer equipped with at
least a PCI card shouldn't be end-user upgradable in both hardware and
OS, as long as you still use the same card.

I used to see this a lot with hardware vendors in the computing world
where one vendor would create a new version of the software which they
claimed required you to use updated hardware- they were full of it.
The new software worked fine on old hardware, and likewise, the old
software worked fine on new hardware.

I think it would be worth an experiment by someone with a slightly
older computer controlled scope to try installing the software on
another machine with a newer OS.

--Justin.

On Wed, Jun 30, 2010 at 4:36 PM, {TindallR-at-missouri.edu} wrote:
}
}
}
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} Dr. B,
}
} I wholeheartedly agree with those who say that the limitation is the computer and software.  We have one instrument running on Windows NT Workstation 4.0.  To upgrade it to an OS from this century would cost an estimated $30-40K (yes, you read that right).
}
} On the other hand, you might still have the old Hitachi S570 running that I started on back in 1987.  The Hitachi HU11's, H500, and probably the H7100 would have run as long as parts and service are available.   And our JEOL 1200EX here was still going after 20+ years of hard use----getting creaky, but still running and producing good results.
}
} I think one of the biggest problems the OEMs should be addressing now is making the software and computer components of their instruments more upgradeable and updatable.  I suspect there are financial as well as technical aspects to this problem, as in more turnover, more profits, but I'll let the OEMs comment on that aspect.
}
} Rando
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway:  http://www.sofn.com
}
}
}
}
} -----Original Message-----
} X-from: bozzola-at-siu.edu [mailto:bozzola-at-siu.edu]
} Sent: Tuesday, June 29, 2010 3:15 PM
} To: Tindall, Randy D.
} Subject: [Microscopy] EM: Life Expectancy of an EM Unit
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} I had a lively discussion recently with an administrator regarding the
} useful life expectancy of an electron beam instrument.
}
} It would be great of such instruments lasted a really long time, but
} they don't.
}
} Here are some questions:
}
} 1.  What has been your personal experience with useable lifetimes of
} electron microscopes (SEM and/or TEM)? Feel free to rant and rave
} (especially if you email me directly).
}
} 2.  Do newer instruments become obsolete at a faster rate than did the
} older instruments? Observations and comments.
}
} To preserve the good will of our sponsors, please email me directly
} (if brand names are given) and I will publish a generic version here.
}
} Thank you.
}
}
}
} --
} John J. Bozzola, Ph.D., Director
} I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
} Southern Illinois University
} 750 Communications Drive
} Carbondale, IL  62901
} Phone: 618-453-3730
}
} ==============================Original Headers==============================
} 10, 16 -- From bozzola-at-siu.edu Tue Jun 29 15:14:20 2010
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} 10, 16 -- Subject: EM: Life Expectancy of an EM Unit
} 10, 16 -- From: John Bozzola {bozzola-at-siu.edu}
} 10, 16 -- To: MSAListserver {Microscopy-at-microscopy.com}
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} ==============================Original Headers==============================
} 24, 36 -- From TindallR-at-missouri.edu Wed Jun 30 09:32:14 2010
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} 24, 36 --  09:32:08 -0500
} 24, 36 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu}
} 24, 36 -- To: "bozzola-at-siu.edu" {bozzola-at-siu.edu}
} 24, 36 -- CC: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
} 24, 36 -- Date: Wed, 30 Jun 2010 09:32:07 -0500
} 24, 36 -- Subject: RE: [Microscopy] EM: Life Expectancy of an EM Unit
} 24, 36 -- Thread-Topic: [Microscopy] EM: Life Expectancy of an EM Unit
} 24, 36 -- Thread-Index: AcsXx8ZOPEXKmqHaRWCSxI11lCqYUwAl53kw
} 24, 36 -- Message-ID: {9422E68616A7C648A281C0B5CD22A4B82A80420797-at-UM-EMAIL06.um.umsystem.edu}
} 24, 36 -- References: {201006292015.o5TKF8t9009228-at-ns.microscopy.com}
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}



--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
Esar


==============================Original Headers==============================
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From: adavison-at-resonantmicro.com
Date: Wed, 30 Jun 2010 10:39:48 -0500
Subject: [Microscopy] SEM

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

We are resurrecting a Hitachi S-510 SEM. The machine is just about fully
functional. We are trying to optimize the filament current but are
having a problem with the waveform modulation. To optimize the current
we focus on a green line on the crt. The width of this green line is
controlled by the contrast knob; we are unable to change this line with
the contrast control. We are trying to determine what sends the signal
to the crt and the contrast control displaying the waveform modulation.
The drawings of the electronic schematics we have are very distorted and
some portions are unreadable. If someone has encountered this or a
similar situation and found a solution to the problem I would appreciate
input?


Aaron Davison
Manufacturing Support Technician
Resonant Microsystems
2322 Alpine Rd
Mailbox #4
Eau Claire, Wi 54703
715-874-6800
adavison-at-resonantmicro.com




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From: jal490-at-nyu.edu
Date: Wed, 30 Jun 2010 14:24:33 -0500
Subject: [Microscopy] Correlative microscopy - registration of fluorescence and EM

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

Dear members.
I'm a researcher doing correlative microscopy. Do you know of a method to correctly register the images gathered from fluorescence microscopy with the ones from electron microscopy. I don't have the exact details but both cameras have a different pixel size, magnifications are different,etc.
Any help would be greatly appreciated.
Thanks in advance.

Jaime Llodra

==============================Original Headers==============================
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From: tina-at-pbrc.hawaii.edu
Date: Wed, 30 Jun 2010 14:45:01 -0500
Subject: [Microscopy] Re: Correlative microscopy - registration of fluorescence

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

Hi, Jaime-

I was browsing for something else and saw this yesterday:

Handbook of Biological Confocal Microscopy, Third edition, edited by James
Pawley, Chapter 49.

Aloha,
Tina



} Dear members.
} I'm a researcher doing correlative microscopy. Do you know of a method to correctly register the images gathered from fluorescence microscopy with the ones from electron microscopy. I don't have the exact details but both cameras have a different pixel size, magnifications are different,etc.
} Any help would be greatly appreciated.
} Thanks in advance.
}
} Jaime Llodra
}
} ==============================Original Headers==============================
} 2, 24 -- From jal490-at-nyu.edu Wed Jun 30 14:24:33 2010
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} 2, 24 -- images
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}

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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From: ZZhang-at-uwyo.edu
Date: Wed, 30 Jun 2010 16:45:27 -0500
Subject: [Microscopy] Confocal Microscope for sale

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

Dear Listservers,
I have a client who wants to improve their GFP fluorescence with microwave fixation. I remember an article on just this subject in Microscopy Today a few years back, does anyone have that reference
or a similar one? Thank you in advance.

Michael Delannoy

==============================Original Headers==============================
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From mitchstyers-at-banzetlaw.com Wed Jun 30 15:04:59 2010
Return-Path: {mitchstyers-at-banzetlaw.com}
Received: from p30174-adsau12honb1-acca.tokyo.ocn.ne.jp (p30174-adsau12honb1-acca.tokyo.ocn.ne.jp [220.98.16.174])
by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id o5UK4uxg032121;
Wed, 30 Jun 2010 15:04:58 -0500
Received: from [220.98.16.174] by mx00-dom.earthlink.net; Thu, 1 Jul 2010 05:04:02 +0900

The University of Wyoming  recently purchased  a new confocal microscope and plans to  sell  its current Leica SP2.  
 
Details of the  LEICA TCS SP2 Laser Scanning Confocal Microscope  are: 

Microscope: Leica DM IRBE inverted microscope with 10X, 20X, 40X, 63X and 100X objectives; 50 W Fluorescence lamp; filters include DAPI, FITC and TRITC.

Laser lines: 458 nm, 488 nm, 543 nm and 633 nm.

Detectors (PMTs): One transmitted light detector and three fluorescence detectors.

The microscope was purchased in  the year 2000 and it is fully functional. It had been under  a  service contract until 2008, but we haven't had any major problem since then. The microscope is as is and with no warranties. The University of Wyoming reserves the right to refuse any and all offers.
 
Please contact me offline if you are interested.

Thank you,

Zhaojie

Zhaojie Zhang, Ph. D.
Director, Microscopy Core Facility
University of Wyoming
Laramie, WY 82071
PHONE: 307-766-3038
FAX: 307-766-5625
http://www.uwyo.edu/microscopy




==============================Original Headers==============================
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From: zaluzec-at-aaem.amc.anl.gov
Date: Wed, 30 Jun 2010 18:04:38 -0500
Subject: [Microscopy] Administrivia: Reminder No "selling" on the Listserver

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

Colleagues

Just a friendly reminder.

Selling items via the Microscopy Listserver is contrary to our
established netiquette &
operating rules. Items can be posted to be given away for free or at
most to cover shipping costs.

I realize that there are ocassions where individuals and/or
organization would like to recoup some costs of items which are still
servicable. That is not the role of this forum.

In such a situation, feel free make use of the Surplus Equipment WWW
site. Here items can be listed for sale, barter or what ever. That
database for that function can be found at

http://www.microscopy.com

Look for the link on the left hand column, called Surplus Equipment.

Remember, if there is any question or doubt just access the FAQ
(http://www.microscopy.com/FAQ.html)
or just send me a private Email message (zaluzec-at-microscopy.com) and
I will be happy to clarify any questions or ambiguities.


Cheers,

Nestor
Your Friendly Neighborhood SysOp


--

===========================================
The box said ...
"This program requires Win 95/98/NT/XP/Vista/Windows 7 or better..."
So I bought a Mac !

===========================================

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15, 24 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov}
15, 24 -- Subject: Administrivia: Reminder No "selling" on the Listserver
15, 24 -- Cc: ZZhang-at-uwyo.edu
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From: paluritamu-at-tamu.edu
Date: Wed, 30 Jun 2010 20:12:24 -0500
Subject: [Microscopy] viaWWW: Microscope Cameras- suggestions

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Email: paluritamu-at-tamu.edu
Name: Rajeshwari

Organization: Texas A&M University

Title-Subject: [Filtered] Microscope Cameras- suggestions

Message: Dear All

I am Rajeshwari, graduate research assistant at Texas A&M University,
USA. I would like some suggestions regarding microscope cameras
(model names or numbers or websites of dealers) for research. What we
need is good resolution (5 MegaPixel) along with a reasonably good
speed(frames/sec) that can capture images and videos and also which
can save the output as raw data (not only as a single video, but as a
bunch of pictures in a file). Our budget is about 500-600USD. We also
prefer a camera with a good software features. Please post your
suggestions.

Thank you
Rajeshwari

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From: mlibbee-at-gmail.com
Date: Wed, 30 Jun 2010 20:12:55 -0500
Subject: [Microscopy] viaWWW: AuPd v. Pt grain size

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Email: mlibbee-at-gmail.com
Name: Marissa Libbee

Title-Subject: [Filtered] AuPd v. Pt grain size

Message: Thanks for the suggestions and comments. Each response is
valuable for the guidance I get on where to look to expand my
understanding and refine my skill set. I am grateful for the advice
on how to improve upon/test my own methods and am forever thankful
that a resource, such as this, exists.



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From: rowland3-at-umbc.edu
Date: Wed, 30 Jun 2010 20:14:37 -0500
Subject: [Microscopy] viaWWW: Imaging In the specimen chamber

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Email: rowland3-at-umbc.edu
Name: Cody Rowland

Organization: Matsys

Title-Subject: [Filtered] Imaging In the specimen chamber

Message: I have a old Jeol JSM-6100 that I'm trying to get running. I
am currently trying to get the infrared chamber scope to feed me a
visible image of the specimen chamber. The infrared camera is a GW
Electronic, type 43. I am not sure how to wire the auto iris to SEM
illumination panel. I called them (and a few other GW electronics)
and got nobody who knew this camera, also I cannot access their
website. I was wondering if anyone knew if they closed down or merged
with another company.

Thanks,
Cody

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From: mckeown3-at-llnl.gov
Date: Wed, 30 Jun 2010 20:15:28 -0500
Subject: [Microscopy] viaWWW: Looking for JEOL 2000 EX/FX microscope

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Email: mckeown3-at-llnl.gov
Name: Joe McKeown

Organization: LLNL

Title-Subject: [Filtered] Looking for JEOL 2000 EX/FX microscope

Message: Hi. I'm looking for a JEOL 200EX or 2000FX TEM, or parts for
this microscope. If anyone is selling/decommissioning one of these
microscopes, or knows of someone who is selling/decommissioning one,
please contact me. It would be much appreciated.

Thanks,

Joe McKeown

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From: gary-at-gaugler.com
Date: Wed, 30 Jun 2010 21:29:48 -0500
Subject: [Microscopy] Re: viaWWW: Microscope Cameras- suggestions

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Good luck. If you find one at this price point,
please let us know.

gary g.


At 06:14 PM 6/30/2010, you wrote:



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From: oshel1pe-at-cmich.edu
Date: Thu, 1 Jul 2010 09:17:07 -0500
Subject: [Microscopy] Re: Preparing figures for publication --PPI vs DPI

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In price range of US$500 - US$600 for professional 5MP video camera made
for microscopy you are, probably, out of luck. Try to look at Nikon
D300/D500/D3000/D5000 camera body and see if it may fit your needs.

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com

On 6/30/2010 9:13 PM, paluritamu-at-tamu.edu wrote:
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} Email: paluritamu-at-tamu.edu
} Name: Rajeshwari
}
} Organization: Texas A&M University
}
} Title-Subject: [Filtered] Microscope Cameras- suggestions
}
} Message: Dear All
}
} I am Rajeshwari, graduate research assistant at Texas A&M University,
} USA. I would like some suggestions regarding microscope cameras
} (model names or numbers or websites of dealers) for research. What we
} need is good resolution (5 MegaPixel) along with a reasonably good
} speed(frames/sec) that can capture images and videos and also which
} can save the output as raw data (not only as a single video, but as a
} bunch of pictures in a file). Our budget is about 500-600USD. We also
} prefer a camera with a good software features. Please post your
} suggestions.
}
} Thank you
} Rajeshwari
}
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From jacobsen-at-terrapins.com Thu Jul 1 00:49:41 2010
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Another editor stepping in ...
First, I have to agree with an earlier point made by Dan White:
digital images are not really images or photographs. They are data
arrays. Tables of numbers that just happen to represent intensity
values at a given (x,y) location on a computer monitor or printed
page. The images we're speaking of are just representations of these
data. (Another way to represent the data is to do a 3D plot, (x,y,z),
where z is the value at point (x,y) in the data file -- a 2D
histogram of the data. An equally valid presentation of the data, but
not an image -- the point being, a digital image is data
presentation, not a picture.)
Doing any image manipulation changes either the data values or the
presentation of the data values. Obviously, anything that changes the
data values is wrong. Changing the presentation can be argued to be
allowable, if how the presentation is changed is made clear.
(Think of any other data table, say, a table of cell survival vs
concentration of a metabolite, and the graph plotted from that table.
Neither can be changed without manipulating the data. Image files are
the same way, we're just (too) used to photomicrographs to have
broken the picture habit.)

Second, I agree completely that there needs to be some depository for
the raw images **with** attached metadata. This latter is very likely
soon to be a matter of law and regulation anyway, at least for
forensic and pharmaceutical related images.
The problem is, who's going to pay for this? How will such a database
(or databases) be maintained, and file formats kept compatible for
years? Not to mention Joel's point about keeping the links to the raw
images valid into the future.

Having said that, we would like to invite someone to write (or more
than one to co-author) an Opinion piece on this topic for publication
in Microscopy Today. If someone would like to do this (don't make me
point), please contact me or Charlie Lyman, MT Editor-in-Chief
(cel1-at-lehigh.edu).

Phil

Folks,

It seems to me that we are talking about purity here. After forty+
years in this biz, I have become quite used to the loss of quality in
printed versions of my micrographs. Once, in the 60's, there was a
French "Journal de Microscopie" that attempted to print high quality
glossies of electron micrographs --beautiful, but unsustainable.
Beyond that, it is clear that there is a significant loss of detail
in printing on paper, regardless of the demands of publishers. --and
forget about dynamic range! The solution that many of us have used is
to show a low magnification image for orientation, with a cutout at
higher magnification to illustrate the fine details. --and forget
about dynamic range!

What I find encouraging is the practice among some journals of
posting online high resolution versions of the low-res images that
they are forced to use in print. This allows one to really examine
the images in detail. Unfortunately, the pdf versions of the papers
generally use the print versions of the images, in order to manage
file sizes, etc. As a result, I often find myself downloading not
only the pdf, but also the detailed figures, for papers where this
matters.

If we want to approach "purity", I would join the chorus of those who
ask publishers to maintain online access to the original high
resolution images that we provide.

In the broader scheme, though, aren't most of our published images
supposed to be representative of a set of experimental results? It
is only very rarely that a single picture representing a single
instance of a phenomenon is worthy of publication. (I recall some
virologists saying "One particle makes an article", but that's a
different story} ) In that case, what we are really after is to
present clarity, so that the reader is able to follow our argument.
Since we are already selecting images for presentation, that is, by
itself, a transform of the original results, and we should be aware
of that.

rant over.

Joel

On Thu, Jul 1, 2010 at 8:31 AM, Guy Cox
{ {mailto:guy.cox-at-sydney.edu.au} guy.cox-at-sydney.edu.au} wrote:

I don't accept that. Read my chapter in the Pawley book. When we
resample, we are trying to get the closest result we can to sampling the
original specimen at the higher resolution. Bicubic interpolation does
that, and is what we should use. We always have to remember that our
digital image is not a precise representation of the microscope image,
it is a set of sample points. Our task is to re-map that set of sample
points to the best possible representation of the image. There are in
principle, I think, even better algorithms for mapping but they are not
likely to be available in your typical image processing software.


Guy

Optical Imaging Techniques in Cell Biology
by Guy Cox CRC Press / Taylor & Francis
{http://www.guycox.com/optical.htm} http://www.guycox.com/optical.htm
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon)
Australian Centre for Microscopy & Microanalysis,
Madsen Building F09, University of Sydney, NSW 2006

Phone +61 2 9351 3176 Fax +61 2 9351 7682
Mobile 0413 281 861
______________________________________________
{http://www.guycox.net} http://www.guycox.net

It would be simpler if journals would specify the exact width(s) of
images that they can accommodate in pixels - you can treat dpi as a
number of pixels.

Making small adjustments to the size of images creates a difference
between the image you submit and the image as published since the need
to increase or decrease the number of pixels involves fabricating new
pixels from the old.

Ideally there should be an integer relationship between the size of
the original images in pixels as it comes off the microscope and the
number of pixels in the submitted image. When magnifying an image by
increasing the number of pixels you should only magnify by an integer
and specify (Photoshop) that interpolation is by using the nearest
neighbour. Then one of your original pixels will still appear to be a
single pixel when printed.
If necessary pad the final image out with a border but don't give the
journal an excuse for fiddling with the number of pixels.

Jeremy Adler
Uppsala U
Sweden

--
Joel B. Sheffield, Ph.D
Department of Biology
Temple University
Philadelphia, PA 19122
Voice: 215 204 8839
e-mail: {mailto:jbs-at-temple.edu} jbs-at-temple.edu
URL: {http://astro.temple.edu/~jbs} http://astro.temple.edu/~jbs


--
Philip Oshel
Technical Editor, Microscopy Today
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576
http://www.microscopy-today.com/

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From: eliceiri-at-wisc.edu
Date: Thu, 1 Jul 2010 09:33:14 -0500
Subject: [Microscopy] Preparing figures for publication --PPI vs DPI

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

with regards to the issue of having access to the original file with metadata with the journal article, it might be of interest to look at the JCB DataViewer project, see
http://jcb-dataviewer.rupress.org/jcb/page/about/

This project from the Open Microscopy Environment (www.openmicroscopy.org) consortium is being used regularly by JCB and allows full viewing of the original microscopy file (uses Bio-Formats to fully parse the proprietary pixel and metadata info). We wrote a recent article on this, see

http://jcb.rupress.org/content/189/5/777.abstract

best,
kevin
}
} On 07/01/10, oshel1pe-at-cmich.edu wrote:
}
} } ----------------------------------------------------------------------------
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} }
} } Another editor stepping in ...
} } First, I have to agree with an earlier point made by Dan White:
} } digital images are not really images or photographs. They are data
} } arrays. Tables of numbers that just happen to represent intensity
} } values at a given (x,y) location on a computer monitor or printed
} } page. The images we're speaking of are just representations of these
} } data. (Another way to represent the data is to do a 3D plot, (x,y,z),
} } where z is the value at point (x,y) in the data file -- a 2D
} } histogram of the data. An equally valid presentation of the data, but
} } not an image -- the point being, a digital image is data
} } presentation, not a picture.)
} } Doing any image manipulation changes either the data values or the
} } presentation of the data values. Obviously, anything that changes the
} } data values is wrong. Changing the presentation can be argued to be
} } allowable, if how the presentation is changed is made clear.
} } (Think of any other data table, say, a table of cell survival vs
} } concentration of a metabolite, and the graph plotted from that table.
} } Neither can be changed without manipulating the data. Image files are
} } the same way, we're just (too) used to photomicrographs to have
} } broken the picture habit.)
} }
} } Second, I agree completely that there needs to be some depository for
} } the raw images **with** attached metadata. This latter is very likely
} } soon to be a matter of law and regulation anyway, at least for
} } forensic and pharmaceutical related images.
} } The problem is, who's going to pay for this? How will such a database
} } (or databases) be maintained, and file formats kept compatible for
} } years? Not to mention Joel's point about keeping the links to the raw
} } images valid into the future.
} }
} } Having said that, we would like to invite someone to write (or more
} } than one to co-author) an Opinion piece on this topic for publication
} } in Microscopy Today. If someone would like to do this (don't make me
} } point), please contact me or Charlie Lyman, MT Editor-in-Chief
} } (cel1-at-lehigh.edu).
} }
} } Phil
} }
} } Folks,
} }
} } It seems to me that we are talking about purity here.  After forty+
} } years in this biz, I have become quite used to the loss of quality in
} } printed versions of my micrographs.  Once, in the 60's, there was a
} } French "Journal de Microscopie" that attempted to print high quality
} } glossies of electron micrographs --beautiful, but unsustainable.
} } Beyond that, it is clear that there is a significant loss of detail
} } in printing on paper, regardless of the demands of publishers. --and
} } forget about dynamic range! The solution that many of us have used is
} } to show a low magnification image for orientation, with a cutout at
} } higher magnification to illustrate the fine details.  --and forget
} } about dynamic range!
} }
} } What I find encouraging is the practice among some journals of
} } posting online high resolution versions of the low-res images that
} } they are forced to use in print.  This allows one to really examine
} } the images in detail.  Unfortunately, the pdf versions of the papers
} } generally use the print versions of the images, in order to manage
} } file sizes, etc.  As a result, I often find myself downloading not
} } only the pdf, but also the detailed figures, for papers where this
} } matters.
} }
} } If we want to approach "purity", I would join the chorus of those who
} } ask publishers to maintain online access to the original high
} } resolution images that we provide.
} }
} } In the broader scheme, though, aren't most of our published images
} } supposed to be representative of a set of experimental results?  It
} } is only very rarely that a single picture representing a single
} } instance of a phenomenon is worthy of publication.  (I recall some
} } virologists saying "One particle makes an article", but that's a
} } different story} )  In that case, what we are really after is to
} } present clarity, so that the reader is able to follow our argument.
} } Since we are already selecting images for presentation, that is, by
} } itself, a transform of the original results, and we should be aware
} } of that.
} }
} } rant over.
} }
} } Joel
} }
} } On Thu, Jul 1, 2010 at 8:31 AM, Guy Cox
} } { {mailto:guy.cox-at-sydney.edu.au(javascript:main.compose()(javascript:main.compose()} guy.cox-at-sydney.edu.au} wrote:
} }
} } I don't accept that.  Read my chapter in the Pawley book.  When we
} } resample, we are trying to get the closest result we can to sampling the
} } original specimen at the higher resolution.  Bicubic interpolation does
} } that, and is what we should use.  We always have to remember that our
} } digital image is not a precise representation of the microscope image,
} } it is a set of sample points.  Our task is to re-map that set of sample
} } points to the best possible representation of the image.  There are in
} } principle, I think, even better algorithms for mapping but they are not
} } likely to be available in your typical image processing software.
} }
} }
} }                                                         Guy
} }
} } Optical Imaging Techniques in Cell Biology
} } by Guy Cox    CRC Press / Taylor & Francis
} }      {http://www.guycox.com/optical.htm} http://www.guycox.com/optical.htm
} } ______________________________________________
} } Associate Professor Guy Cox, MA, DPhil(Oxon)
} } Australian Centre for Microscopy & Microanalysis,
} } Madsen Building F09, University of Sydney, NSW 2006
} }
} } Phone +61 2 9351 3176     Fax +61 2 9351 7682
} }              Mobile 0413 281 861
} } ______________________________________________
} }       {http://www.guycox.net} http://www.guycox.net
} }
} } It would be simpler if journals would specify the exact width(s) of
} } images that they can accommodate in pixels - you can treat dpi as a
} } number of pixels.
} }
} } Making small adjustments to the size of images creates a difference
} } between the image you submit and the image as published since the need
} } to increase or decrease the number of pixels involves fabricating new
} } pixels from the old.
} }
} } Ideally there should be an integer relationship between the size of
} } the original images in pixels as it comes off the microscope and the
} } number of pixels in the submitted image. When magnifying an image by
} } increasing the number of pixels you should only magnify by an integer
} } and specify (Photoshop) that interpolation is by using the nearest
} } neighbour. Then one of your original pixels will  still appear to be a
} } single pixel when printed.
} } If necessary pad the final image out with a border but don't give the
} } journal an excuse for fiddling with the number of pixels.
} }
} } Jeremy Adler
} } Uppsala U
} } Sweden
} }
} } --
} } Joel B. Sheffield, Ph.D
} } Department of Biology
} } Temple University
} } Philadelphia, PA 19122
} } Voice: 215 204 8839
} } e-mail: {mailto:jbs-at-temple.edu(javascript:main.compose()(javascript:main.compose()} jbs-at-temple.edu
} } URL:  {http://astro.temple.edu/~jbs} http://astro.temple.edu/~jbs
} }
} }
} } --
} } Philip Oshel
} } Technical Editor, Microscopy Today
} } Microscopy Facility Supervisor
} } Biology Department
} } 024C Brooks Hall
} } Central Michigan University
} } Mt. Pleasant, MI 48859
} } (989) 774-3576
} } http://www.microscopy-today.com/
} }  
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--
Kevin W. Eliceiri
Director
Laboratory for Optical and Computational Instrumentation
http://www.loci.wisc.edu
Room 271 Animal Sciences
1675 Observatory Drive
Madison, WI 53706
Phone: 608-263-6288
Fax: 608-262-4570


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From: jae5-at-lehigh.edu
Date: Thu, 1 Jul 2010 10:37:04 -0500
Subject: [Microscopy] Electron optics of thermionic electron guns

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

This is a question about thermionic electron guns in scanning and
transmission electron microscopes. The question is about tungsten
filaments or LaB6 filaments (not about field emission or Schottky sources).

To illustrate the function of the electron gun, standard textbooks
(Reimer; Goldstein et al; De Graaf; Williams and Carter) all show
diagrams with a cross over between the filament and the anode.
Somewhere along the way, I have acquired the impression that real
microscopes do not work this way.

I have long believed (though I can not recall where the belief comes
from) that the manufacturers design the gun so that there is no cross
over in front of the anode. I thought that the gun was designed to
operate with a virtual source just behind the filament tip. With no
real cross over until below the condenser lens. The point being that,
if there were a cross over where the electrons are traveling rather
slowly and before an aperture has removed a lot of the beam, the Boersch
effect would increase the energy spread unnecessarily.

Am I right? Is this way it is done in practice or are the books right?
I would greatly appreciate an answer to this question from someone who
really knows.

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


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From: hkonishi-at-wisc.edu
Date: Thu, 1 Jul 2010 10:43:58 -0500
Subject: [Microscopy] Off line calibration of ED distortion

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

Hello,
I would like to know if there is a software that can calibrate the distortion of electron diffraction patterns that were recorded by Digital Micrograph. I want to calibrate the distortion in all the directions from the central spot. I will use a ring pattern (Au) for a standard.

Thank you,
Hiromi Konishi
UW-Madison


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From: kenconverse-at-qualityimages.biz
Date: Thu, 1 Jul 2010 11:30:56 -0500
Subject: [Microscopy] viaWWW: Imaging In the specimen chamber

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

Hi Cody,
GW Electronics was bought a number of years ago by K. E. Developments. They
can be found at
http://www.kedev.co.uk/
or contact Graham Wardall [gwardall-at-kedev.com].

I ran across one of these recently and wasn't able to get any documentation
on it but you might try contacting Vincent Difilippo at Agiltron in Woburn,
MA [vdifilippo-at-agiltron.com]. He has a Type 43, that works, on a Hitachi
S-2700.

Ken Converse
owner


QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
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Email: rowland3-at-umbc.edu
Name: Cody Rowland

Organization: Matsys

Title-Subject: [Filtered] Imaging In the specimen chamber

Message: I have a old Jeol JSM-6100 that I'm trying to get running. I
am currently trying to get the infrared chamber scope to feed me a
visible image of the specimen chamber. The infrared camera is a GW
Electronic, type 43. I am not sure how to wire the auto iris to SEM
illumination panel. I called them (and a few other GW electronics)
and got nobody who knew this camera, also I cannot access their
website. I was wondering if anyone knew if they closed down or merged
with another company.

Thanks,
Cody

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From: mike.bode-at-resaltatech.com
Date: Thu, 1 Jul 2010 12:18:28 -0500
Subject: [Microscopy] Preparing figures for publication --PPI vs DPI

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

Hi,

I think that this is the approach being taken by SemTech Solutions (Billerica, MA), which makes much of its living updating Amray SEMs. You might give them a shout about expanding to other brands and models.

Good hunting,
Barbara Foster, President and Sr. Consultant

Microscopy/Microscopy Education
7101 Royal Glen Trail, Suite A
McKinney TX 75070
P: (972)924-5310 Skype: fostermme
W: www.MicroscopyEducation.com

NEWS! Visit the NEW and IMPROVED www.MicroscopyEducation.com! And don't forget: MME is now scheduling customized, on-site courses for the Summer and Fall. Call me for a free assessment and quote.


At 10:57 AM 6/30/2010, kraftpiano-at-gmail.com wrote:



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From kefxebecllcdiz-at-xebecllc.com Thu Jul 1 11:52:35 2010
Return-Path: {kefxebecllcdiz-at-xebecllc.com}
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As someone who has been in the "imaging business" for a while, I'd like to
comment on this thread. I think there are at least 2 issues that are being
discussed here:

1) publishing images in journals
2) dealing with meta data

Regarding issue 1, I don't really see a big issue, but perhaps I am
mistaken. When I published, I never thought of the images in the publication
as anything more than visual data to prove a point, which would be described
in the text. I never thought that someone would look at the image in the
magazine at the pixel level and try to draw any conclusions from that. Given
that, a resampling of the image using a bi-cubic spline (or other) doesn't
really matter that much (provided no artificial artifacts like Moire are
introduced). It's even easier for online publications: provide a link to the
original data. Am I totally off the mark here? I mean, if you need to show
data at the pixel level, I would not rely on people using a magnifying glass
to see the data.

Dealing with meta data is a more complex issue. There are standards (TIF,
for example) that allow meta data to be stored with the image in a (sort of)
standardized way. The problem is that microscopy is a tiny little fraction
of the imaging world, and the standard bearers are not focused on microscopy
(no pun intended). For example, TIF has a tag for "Resolution". The problem
is that it is used differently from the way a microscopist would use it. It
is essentially the print resolution. So, if you specify that as, say, 10-6,
or a micron, a program such as Word will try to print it at that resolution.
A 1kx1k image would simply print out as 1mm x 1mm. Not what is intended.
There are really no standard microscopy tags in the TIF definition. That
leaves the other option: use private tags. That works fine as long as you
stay in the same software, but software from another manufacturer cannot
read the tags, so there is no interoperability.

With the microscopy community so small (relatively speaking), I think it is
up to us (perhaps MSA?) to define some internal standards and possibly try
to get them embedded in a general standard. Even if it the acceptance within
the TIF (or other) standard is a long shot, if MSA would publish a list of
keywords and tags to be used, I think that most software manufacturers would
quickly adopt it and at least provide interoperability among microscopy
software. Most manufacturers of cameras and software, for example, collect a
whole range of data (camera data, microscope data, imaging conditions, etc.)
which are then stored somehow (sometimes in private tiff tags, others use a
separate text file, etc.). the problem is getting everybody on the same
page.

Mike Bode
---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com








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From: mike.bode-at-resaltatech.com
Date: Thu, 1 Jul 2010 12:40:45 -0500
Subject: [Microscopy] EM: Life Expectancy of an EM Unit

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

Frankly, I don't think it is that easy. Computers and Operating systems
evolve, and it is not always possible to simply install software on a new
OS. If you are talking just about software, I think it might be possible,
although even there I have my doubts. Hardware is where the problem lies.
Let's simply take a look at the standards for add-on cards: ISA, EISA, PCI,
PCI-E. None of these is backward compatible, and you can't find any PCs with
the first 2 anymore. Even PCI is hard to find nowadays.
Now, remember, the cards that you find to control a microscope are most
likely custom made cards, and a microscope manufacturer will likely not
re-develop a PCI-E card for a customer who needs some functionality that
they bought 15 years ago on an EISA card. Most people will not be able to do
that themselves, so they are stuck: buy a new microscope, or pay the
microscope manufacturer for their help. You may balk at the prices they
quote you, but they (the column manufacturers) either have to embark on a
re-development, which can take a lot of man-hours, or they had the foresight
to stock some of the older computers and keep them and the software in
storage for decades in case someone needs a computer in the future. This is
not a cheap option for the manufacturers, so don't be surprised at the price
tag.

TEMs are driven by technology just as much as they are by user demand. That
means automation and capabilities that are unthinkable without the help of
computers. But computers are driven by a completely different set of demands
(Remember Moore's Law?). And that is what causes the problem, I think.

For the user, I think there is something they can do: First, keep the
computer that runs the microscope as lean as possible. Don't weigh it down
with applications like word processing or other applications. If the
computer runs the microscope adequately, then it will do so still in 10
years. General computing may have surpassed the computer and OS a long time
ago, but it will run the microscope just as well as it did the first day.
Use a second computer to run software that is not necessary for running the
microscope itself. Some manufacturers do that anyway.
And third, if you are really concerned about the microscope computer
breaking down: Buy a second, identical computer and put it in storage. If
the first computer breaks, transfer the cards and hard disk, and you should
be up again in no time. If down the road one of the cards breaks, you only
need to find a replacement, which is a lot easier than having the board and
software re-engineered for a different standard.

These are just my 2 cents.

Mike Bode
---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com






-----Original Message-----
X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com]
Sent: Wednesday, June 30, 2010 9:10 AM
To: mike.bode-at-resaltatech.com

I've been reading this thread with some interest, as in a previous
life, I was a computer consultant (Although I don't like admitting it
in public.) There are ways to make older software run on newer
operating systems and computers, and I wonder if the marriage between
the EM software and the computer is more of an artificial one.

For example, I received a JEOL DSG unit to use with the 35 that was
installed where I was teaching, and despite the software being written
for windows 3.1, it is now working perfectly on windows XP. The only
limitation that I've found with that system is that the hardware is an
old ISA board, but I'm sure that with the proper schematics (Which I
have) and time (Which I don't have) I could re-wire it for a more
modern PCI card and get similar results without too much hassle.

I don't see any reason why a modern EM, computer equipped with at
least a PCI card shouldn't be end-user upgradable in both hardware and
OS, as long as you still use the same card.

I used to see this a lot with hardware vendors in the computing world
where one vendor would create a new version of the software which they
claimed required you to use updated hardware- they were full of it.
The new software worked fine on old hardware, and likewise, the old
software worked fine on new hardware.

I think it would be worth an experiment by someone with a slightly
older computer controlled scope to try installing the software on
another machine with a newer OS.

--Justin.



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From: marisa_ote-at-yahoo.com
Date: Thu, 1 Jul 2010 12:43:25 -0500
Subject: [Microscopy] UV bulb replacement for Leica AFS1

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

Hello,

I just heard from my local Leica sales representative that Leica does not provide customers with UV bulbs for the AFS1 anymore. The only available option is to buy a new attachment (a modified version of the UV LED attachment of the newer AFS2) for $2,200!!!! Does anyone know about other suppliers selling this type of UV bulb or a cheaper solution that does not involved spending $2,200? Thanks

Marisa

Marisa Otegui
Department of Botany
University of Wisconsin-Madison





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From: smithj-at-winthrop.edu
Date: Thu, 1 Jul 2010 13:06:15 -0500
Subject: [Microscopy] EM: Life Expectancy of an EM Unit

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

All good advice.
The additional problem we've run into is that the "control" computer
eventually falls off of our IT/Networking dept's radar. We have two
instruments (an SEM and a gel imager) for which we have to transfer
files using Zip disks--neither computer is compatible with our network
infrastructure.
Julian

mike.bode-at-resaltatech.com wrote:
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Frankly, I don't think it is that easy. Computers and Operating systems
} evolve, and it is not always possible to simply install software on a new
} OS. If you are talking just about software, I think it might be possible,
} although even there I have my doubts. Hardware is where the problem lies.
} Let's simply take a look at the standards for add-on cards: ISA, EISA, PCI,
} PCI-E. None of these is backward compatible, and you can't find any PCs with
} the first 2 anymore. Even PCI is hard to find nowadays.
} Now, remember, the cards that you find to control a microscope are most
} likely custom made cards, and a microscope manufacturer will likely not
} re-develop a PCI-E card for a customer who needs some functionality that
} they bought 15 years ago on an EISA card. Most people will not be able to do
} that themselves, so they are stuck: buy a new microscope, or pay the
} microscope manufacturer for their help. You may balk at the prices they
} quote you, but they (the column manufacturers) either have to embark on a
} re-development, which can take a lot of man-hours, or they had the foresight
} to stock some of the older computers and keep them and the software in
} storage for decades in case someone needs a computer in the future. This is
} not a cheap option for the manufacturers, so don't be surprised at the price
} tag.
}
} TEMs are driven by technology just as much as they are by user demand. That
} means automation and capabilities that are unthinkable without the help of
} computers. But computers are driven by a completely different set of demands
} (Remember Moore's Law?). And that is what causes the problem, I think.
}
} For the user, I think there is something they can do: First, keep the
} computer that runs the microscope as lean as possible. Don't weigh it down
} with applications like word processing or other applications. If the
} computer runs the microscope adequately, then it will do so still in 10
} years. General computing may have surpassed the computer and OS a long time
} ago, but it will run the microscope just as well as it did the first day.
} Use a second computer to run software that is not necessary for running the
} microscope itself. Some manufacturers do that anyway.
} And third, if you are really concerned about the microscope computer
} breaking down: Buy a second, identical computer and put it in storage. If
} the first computer breaks, transfer the cards and hard disk, and you should
} be up again in no time. If down the road one of the cards breaks, you only
} need to find a replacement, which is a lot easier than having the board and
} software re-engineered for a different standard.
}
} These are just my 2 cents.
}
} Mike Bode
} ---
} Mike Bode, Ph.D.
} ResAlta Research Technologies Corp.
}
} 2102 Beech Ct
} Golden, CO 80401
} USA
}
} p: +1-303-748-4346
} f: +1-303-202-6350
} Mike.Bode-at-ResAltaTech.com
} www.ResAltaTech.com
}
}
}
}
}
}
} -----Original Message-----
} X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com]
} Sent: Wednesday, June 30, 2010 9:10 AM
} To: mike.bode-at-resaltatech.com
} Subject: [Microscopy] EM: Life Expectancy of an EM Unit
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} I've been reading this thread with some interest, as in a previous
} life, I was a computer consultant (Although I don't like admitting it
} in public.) There are ways to make older software run on newer
} operating systems and computers, and I wonder if the marriage between
} the EM software and the computer is more of an artificial one.
}
} For example, I received a JEOL DSG unit to use with the 35 that was
} installed where I was teaching, and despite the software being written
} for windows 3.1, it is now working perfectly on windows XP. The only
} limitation that I've found with that system is that the hardware is an
} old ISA board, but I'm sure that with the proper schematics (Which I
} have) and time (Which I don't have) I could re-wire it for a more
} modern PCI card and get similar results without too much hassle.
}
} I don't see any reason why a modern EM, computer equipped with at
} least a PCI card shouldn't be end-user upgradable in both hardware and
} OS, as long as you still use the same card.
}
} I used to see this a lot with hardware vendors in the computing world
} where one vendor would create a new version of the software which they
} claimed required you to use updated hardware- they were full of it.
} The new software worked fine on old hardware, and likewise, the old
} software worked fine on new hardware.
}
} I think it would be worth an experiment by someone with a slightly
} older computer controlled scope to try installing the software on
} another machine with a newer OS.
}
} --Justin.
}
}
}
} ==============================Original Headers==============================
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} 25, 31 -- Subject: RE: [Microscopy] EM: Life Expectancy of an EM Unit
} 25, 31 -- Date: Thu, 1 Jul 2010 11:40:29 -0600
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}


--
Julian P.S. Smith III
Director, Winthrop Microscopy Facility
Dept. of Biology
Winthrop University
520 Cherry Rd.
Rock Hill, SC 29733

803-323-2111 x6427 (vox)
803-323-3448 (fax)
803-524-2347 (cell)


==============================Original Headers==============================
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From: PhillipsT-at-missouri.edu
Date: Thu, 1 Jul 2010 13:11:39 -0500
Subject: [Microscopy] EM: Life Expectancy of an EM Unit

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

Sure a PC limits the life of a EM but at least you don't have to buy a whole new one when the light bulb burns out (for those of you who saw Marisa Otegui's recent posting on her Leica AFS) so stop bitching! Tom


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: smithj-at-winthrop.edu [mailto:smithj-at-winthrop.edu]
Sent: Thursday, July 01, 2010 1:07 PM
To: Phillips, Thomas E.

All good advice.
The additional problem we've run into is that the "control" computer
eventually falls off of our IT/Networking dept's radar. We have two
instruments (an SEM and a gel imager) for which we have to transfer
files using Zip disks--neither computer is compatible with our network
infrastructure.
Julian

mike.bode-at-resaltatech.com wrote:
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Frankly, I don't think it is that easy. Computers and Operating systems
} evolve, and it is not always possible to simply install software on a new
} OS. If you are talking just about software, I think it might be possible,
} although even there I have my doubts. Hardware is where the problem lies.
} Let's simply take a look at the standards for add-on cards: ISA, EISA, PCI,
} PCI-E. None of these is backward compatible, and you can't find any PCs with
} the first 2 anymore. Even PCI is hard to find nowadays.
} Now, remember, the cards that you find to control a microscope are most
} likely custom made cards, and a microscope manufacturer will likely not
} re-develop a PCI-E card for a customer who needs some functionality that
} they bought 15 years ago on an EISA card. Most people will not be able to do
} that themselves, so they are stuck: buy a new microscope, or pay the
} microscope manufacturer for their help. You may balk at the prices they
} quote you, but they (the column manufacturers) either have to embark on a
} re-development, which can take a lot of man-hours, or they had the foresight
} to stock some of the older computers and keep them and the software in
} storage for decades in case someone needs a computer in the future. This is
} not a cheap option for the manufacturers, so don't be surprised at the price
} tag.
}
} TEMs are driven by technology just as much as they are by user demand. That
} means automation and capabilities that are unthinkable without the help of
} computers. But computers are driven by a completely different set of demands
} (Remember Moore's Law?). And that is what causes the problem, I think.
}
} For the user, I think there is something they can do: First, keep the
} computer that runs the microscope as lean as possible. Don't weigh it down
} with applications like word processing or other applications. If the
} computer runs the microscope adequately, then it will do so still in 10
} years. General computing may have surpassed the computer and OS a long time
} ago, but it will run the microscope just as well as it did the first day.
} Use a second computer to run software that is not necessary for running the
} microscope itself. Some manufacturers do that anyway.
} And third, if you are really concerned about the microscope computer
} breaking down: Buy a second, identical computer and put it in storage. If
} the first computer breaks, transfer the cards and hard disk, and you should
} be up again in no time. If down the road one of the cards breaks, you only
} need to find a replacement, which is a lot easier than having the board and
} software re-engineered for a different standard.
}
} These are just my 2 cents.
}
} Mike Bode
} ---
} Mike Bode, Ph.D.
} ResAlta Research Technologies Corp.
}
} 2102 Beech Ct
} Golden, CO 80401
} USA
}
} p: +1-303-748-4346
} f: +1-303-202-6350
} Mike.Bode-at-ResAltaTech.com
} www.ResAltaTech.com
}
}
}
}
}
}
} -----Original Message-----
} X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com]
} Sent: Wednesday, June 30, 2010 9:10 AM
} To: mike.bode-at-resaltatech.com
} Subject: [Microscopy] EM: Life Expectancy of an EM Unit
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} I've been reading this thread with some interest, as in a previous
} life, I was a computer consultant (Although I don't like admitting it
} in public.) There are ways to make older software run on newer
} operating systems and computers, and I wonder if the marriage between
} the EM software and the computer is more of an artificial one.
}
} For example, I received a JEOL DSG unit to use with the 35 that was
} installed where I was teaching, and despite the software being written
} for windows 3.1, it is now working perfectly on windows XP. The only
} limitation that I've found with that system is that the hardware is an
} old ISA board, but I'm sure that with the proper schematics (Which I
} have) and time (Which I don't have) I could re-wire it for a more
} modern PCI card and get similar results without too much hassle.
}
} I don't see any reason why a modern EM, computer equipped with at
} least a PCI card shouldn't be end-user upgradable in both hardware and
} OS, as long as you still use the same card.
}
} I used to see this a lot with hardware vendors in the computing world
} where one vendor would create a new version of the software which they
} claimed required you to use updated hardware- they were full of it.
} The new software worked fine on old hardware, and likewise, the old
} software worked fine on new hardware.
}
} I think it would be worth an experiment by someone with a slightly
} older computer controlled scope to try installing the software on
} another machine with a newer OS.
}
} --Justin.
}
}
}
} ==============================Original Headers==============================
} 25, 31 -- From mike.bode-at-resaltatech.com Thu Jul 1 12:40:44 2010
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--
Julian P.S. Smith III
Director, Winthrop Microscopy Facility
Dept. of Biology
Winthrop University
520 Cherry Rd.
Rock Hill, SC 29733

803-323-2111 x6427 (vox)
803-323-3448 (fax)
803-524-2347 (cell)


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From: drg.mitchell-at-sydney.edu.au
Date: Thu, 1 Jul 2010 19:42:03 -0500
Subject: [Microscopy] viaWWW: Off line calibration of ED distortion

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

Please respond directly to DJ Alwatter and not to the list.

X-from: DJ Alwattar [mailto:djalw-at-northlandlabs.com]
Sent: Monday, June 28, 2010 2:52 PM
To: AssociationManagement

Hello everyone,

Just to add a comment to one of Phil's points a few messages ago, there is a repository in progress; The Cell: An Image Library is a new project of the American Society for Cell Biology (ASCB) and is funded through an NIH/NIGMS grant. And we need your images.

We also use the Open Microscopy Environment platform and Bio-Formats which allow us to accept 75 different formats for the repository. The microscopy data submitted to The Cell will be made available to the research community and the public as public domain works. So while you may present a single image in a journal, we are very interested in making available all the "sister" images you generated in your research. The value to others could be immeasurable. The data in the website will be available to download either in the OME-TIF format or in the format of the original submission, so that researchers can download and analyze the data as they choose.

Please visit www.cellimagelibrary.org for more information and see www.cellimagelibrary.org/participate to learn how to submit images. (It's easy.)

Please do remember though, that if you are going to be publishing an image, you would not want to submit it to The Cell. Unfortunately, many publishers would consider this prior publication. The sister images, however, can be submitted to The Cell.

For more information of with any questions, please contact David Orloff, dorloff-at-ascb.org, Manager, Image Library at the ASCB.

David




-----Original Message-----
X-from: mike.bode-at-resaltatech.com [mailto:mike.bode-at-resaltatech.com]
Sent: Thursday, July 01, 2010 1:26 PM
To: David Orloff

As someone who has been in the "imaging business" for a while, I'd like to
comment on this thread. I think there are at least 2 issues that are being
discussed here:

1) publishing images in journals
2) dealing with meta data

Regarding issue 1, I don't really see a big issue, but perhaps I am
mistaken. When I published, I never thought of the images in the publication
as anything more than visual data to prove a point, which would be described
in the text. I never thought that someone would look at the image in the
magazine at the pixel level and try to draw any conclusions from that. Given
that, a resampling of the image using a bi-cubic spline (or other) doesn't
really matter that much (provided no artificial artifacts like Moire are
introduced). It's even easier for online publications: provide a link to the
original data. Am I totally off the mark here? I mean, if you need to show
data at the pixel level, I would not rely on people using a magnifying glass
to see the data.

Dealing with meta data is a more complex issue. There are standards (TIF,
for example) that allow meta data to be stored with the image in a (sort of)
standardized way. The problem is that microscopy is a tiny little fraction
of the imaging world, and the standard bearers are not focused on microscopy
(no pun intended). For example, TIF has a tag for "Resolution". The problem
is that it is used differently from the way a microscopist would use it. It
is essentially the print resolution. So, if you specify that as, say, 10-6,
or a micron, a program such as Word will try to print it at that resolution.
A 1kx1k image would simply print out as 1mm x 1mm. Not what is intended.
There are really no standard microscopy tags in the TIF definition. That
leaves the other option: use private tags. That works fine as long as you
stay in the same software, but software from another manufacturer cannot
read the tags, so there is no interoperability.

With the microscopy community so small (relatively speaking), I think it is
up to us (perhaps MSA?) to define some internal standards and possibly try
to get them embedded in a general standard. Even if it the acceptance within
the TIF (or other) standard is a long shot, if MSA would publish a list of
keywords and tags to be used, I think that most software manufacturers would
quickly adopt it and at least provide interoperability among microscopy
software. Most manufacturers of cameras and software, for example, collect a
whole range of data (camera data, microscope data, imaging conditions, etc.)
which are then stored somehow (sometimes in private tiff tags, others use a
separate text file, etc.). the problem is getting everybody on the same
page.

Mike Bode
---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com








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Email: drg.mitchell-at-sydney.edu.au
Name: David Mitchell

Organization: University of Sydney

Title-Subject: [Microscopy] Off line calibration of ED distortion

Message: Dear Hiromi

Capitani et al. wrote an interesting paper on the topic of
diffraction pattern distortion (Ultramicroscopy 106 (2006) 64-74).
They measured the distortion using an Excel spreadsheet. You might
contact the authors and see if they will share their spreadsheet with
you. I am unaware of a DigitalMicrograph-based solution for this
measurement. That said it might make an interesting addition to
DiffTools (www.dmscripting.com/difftools). I'll have a think about
developing one. For simply estimating whether visible distortion is
present you might experiment with the 'Locate SADP Centre' tool which
overlays a series of rings on the pattern. The CHT Diffraction
Analysis tool (available from the above web site) can also be used to
find the centre of a polycrystalline pattern and if elliptical
distortion is present in the diffraction rings, then this will be
very evident in the Hough transform images. Unfortunately, it does
not quantify the distortion.

Regards, Dave Mitchell

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From: microwink-at-gmail.com
Date: Thu, 1 Jul 2010 20:07:53 -0500
Subject: [Microscopy] Re: Off line calibration of ED distortion

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

Hi Hiromi,

I found a script on the Digital MicroGraph Script Database hosted by
the Graz University of Technology that does what you are asking. The
link to the script to calculate distortions in the diffraction pattern
introduced by the projector lens stack is here:

http://felmpc14.tu-graz.ac.at/dm_scripts/freeware/programs/Diffraction-Rings-Distortion-Analysis.htm

A link to the database is here:
http://www.felmi-zfe.tugraz.at/dm_scripts/dm_scripts/

The summary of the script's function is as follows: "This is a script
package to characterize diffraction ring distortion in TEM. Due to the
distortion introduced by the TEM projection lens, diffraction rings
obtained from a fine crystalline sample are of elliptical shape
instead of round." I have no connection to this script nor have I
tried it so I can't comment on its function, but it seems to do what
you want. There are many other scripts in the database should this one
not be what you're looking for.

Cheers,
Chris

Christopher Winkler
Dynamic Characterization Group
http://www.materials.drexel.edu/dcg/
Drexel University
267-496-0587


} On Thu, Jul 1, 2010 at 11:50 AM, {hkonishi-at-wisc.edu} wrote:
} }
} }
} }
} }
} } ----------------------------------------------------------------------------
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} }
} } Hello,
} } I would like to know if there is a software that can calibrate the
} } distortion of electron diffraction patterns that were recorded by Digital
} } Micrograph. I want to calibrate the distortion in all the directions from
} } the central spot. I will use a ring pattern (Au) for a standard.
} }
} } Thank you,
} } Hiromi Konishi
} } UW-Madison
} }
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} } 3, 34 -- From: HIROMI KONISHI {hkonishi-at-wisc.edu}
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} --
} Christopher Winkler
} Dynamic Characterization Group
} http://www.materials.drexel.edu/dcg/
} Drexel University
} 267-496-0587
}



--
Christopher Winkler
Dynamic Characterization Group
http://www.materials.drexel.edu/dcg/
Drexel University
267-496-0587


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From: bigelow-at-umich.edu
Date: Thu, 1 Jul 2010 22:00:29 -0500
Subject: [Microscopy] RE: Electron gun optics

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

Haine, et. al. carried out the defining studies of the
characteristics of the biased electron gun in a series of papers
published in the 1950s. These studies are nicely summarized in
Chapter 6 of Haine's book, The Electron Microscope (Interscience
Publishers, 1961).

In discussing the establishment of the saturation condition on page
131, Haine states "Farther out there is a strong radially inward
component to the field, which acts on the relatively slow electrons
and guides them across the axis. This radial field increases rapidly
with radius having a large component increasing with the cube of the
radius, that acts like a fourth order plate in close proximity to the
cathode. The electron rays produce a caustic in front of the cathode,
and somewhere in their image is a minimum cross section. This is the
virtual source." Also, " Haine shows plots of this focusing
arrangement on page 130, and gives a plot of "Cathode Image
Position" in Fig. 6.7 on page 127. Later, while discussing the
design of the illumination system, Haine presents a diagram (Fig.
8.3, page 162) that clearly shows that he believes there is a
crossover outside the grid cap that serves as the source in the
image-forming system. Although I have not operated an electron
microscope for a number of years now, I seem to remember that when
going from the image of the tip of the filament with its surrounding
halo to the final beam spot, it is necessary to increase the strength
of the condenser lens slightly to bring the final beam spot into good
focus. This would also indicate that the final spot is located
closer to the condenser lens than the filament.

Concerning space charge effects, on pages 129 and 130 Haine states,
"From the experimental results already described, and other
considerations, it is possible to deduce a reasonably accurate
picture of the mechanism of beam formation in the electron gun. The
experiments show that, within the useable range of cathode
temperatures, space charge has negligible effect on the beam which,
therefore, is determined only by geometric considerations".Also, on
page 127 he states, "The fact that beam divergence angle is
independent of temperature shows that no beam divergence due to space
charge effects is present, Thus, contrary to all previous belief,
space charge effects were proved to be unimportant in this type of
electron gun".

Now for some reservations. In most of their experiments Haine and
his coauthors used a flat-faced grid cap with a hole that ranged from
0.76 to 2.3 mm in diameter, and "Vee"-shaped hairpin filaments made
of 0.005 inch (0,127 mm) diameter tungsten wire that were located
from 0.25 to 1.0 mm behind the face of the grid cap.. I believe that
most modern guns have conical grid caps, a geometry that was not
investigated by Haine, and so it is remotely possible that the
mechanism of beam formation in these guns is different from that
described by Haine. It would therefore be very interesting to hear
from someone who has had direct experience with the design and
characterization of these modern electron guns.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From: protrain-at-emcourses.com
Date: Fri, 2 Jul 2010 05:02:10 -0500
Subject: [Microscopy] Electron optics of thermionic electron guns

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

Hi Alwyn,

I think you are confusing field emission, where the virtual source is in the
tip, with thermionic emission from a biased electron gun, where the virtual
source is between the cathode and the anode.

Following on from Prof Bigelow's comment, in the dim and distant past I had
a conical cathode cap made for a Hitachi instrument that had a flat cathode
cap; I could not see any improvement in the quality of the source.

The anode to cathode distance plays a big part in the creation of the
virtual source, which would further indicate that the source is between
these components. Lowering the accelerating voltage from its maximum weakens
this field allowing the source to expand in size which contributes to the
fall off in resolution particularly in the SEM. 1970s Hitachi and Philips
TEM had variable anode height adjustable either automatically (Hitachi 1mm
for every 4kV) or externally (Philips). Philips introduced a low kV anode
and cathode at one time, whilst Cambridge/Leo/Zeiss have had available a low
kV anode for many years. I have always encouraged operators of SEM to take
advantage of this area of the gun, trying to work near to the "optimum" SEM
spacing of 1mm for every 2kV by raising the anode. I once modified a JEOL
840 with a Lab6 source such that we could only run safely (beware of
discharge) up to 5kV. However the peak performance was achieved at 4.5kV
whilst we used the instrument a great deal at 200v, yes volts

X-from my 46 years playing with biased electron guns I am totally convinced
that the virtual source is between the anode and cathode!

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



X-from: jae5-at-lehigh.edu [mailto:jae5-at-lehigh.edu]
Sent: 01 July 2010 16:38
To: protrain-at-emcourses.com

This is a question about thermionic electron guns in scanning and
transmission electron microscopes. The question is about tungsten
filaments or LaB6 filaments (not about field emission or Schottky sources).

To illustrate the function of the electron gun, standard textbooks
(Reimer; Goldstein et al; De Graaf; Williams and Carter) all show
diagrams with a cross over between the filament and the anode.
Somewhere along the way, I have acquired the impression that real
microscopes do not work this way.

I have long believed (though I can not recall where the belief comes
from) that the manufacturers design the gun so that there is no cross
over in front of the anode. I thought that the gun was designed to
operate with a virtual source just behind the filament tip. With no
real cross over until below the condenser lens. The point being that,
if there were a cross over where the electrons are traveling rather
slowly and before an aperture has removed a lot of the beam, the Boersch
effect would increase the energy spread unnecessarily.

Am I right? Is this way it is done in practice or are the books right?
I would greatly appreciate an answer to this question from someone who
really knows.

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


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From: nizets2-at-yahoo.com
Date: Fri, 2 Jul 2010 05:38:53 -0500
Subject: [Microscopy] EM: Life Expectancy of an EM Unit

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

I completely agree with that. I think it is a mistake to buy a 1 million $ worth machine and think that you won't need any additional expenses during the next 20 years. When you buy the microscope (and not 5-10 years later), it is wise to calculate the financing of the necessary updates.
When you have a budget for 1 Million $ (or half of it if you like), it seems reasonable to keep 30-40 k$ for future upgrades. Or as Mike said, just buy 3 identical computers and you have enough harware at your disposal. Preventing instead of curing has always been a winner strategy ;-)

Stephane

 


----- Original Message ----
X-from: "mike.bode-at-resaltatech.com" {mike.bode-at-resaltatech.com}
To: nizets2-at-yahoo.com
Sent: Thu, July 1, 2010 7:44:09 PM

Frankly, I don't think it is that easy. Computers and Operating systems
evolve, and it is not always possible to simply install software on a new
OS. If you are talking just about software, I think it might be possible,
although even there I have my doubts. Hardware is where the problem lies.
Let's simply take a look at the standards for add-on cards: ISA, EISA, PCI,
PCI-E. None of these is backward compatible, and you can't find any PCs with
the first 2 anymore. Even PCI is hard to find nowadays.
Now, remember, the cards that you find to control a microscope are most
likely custom made cards, and a microscope manufacturer will likely not
re-develop a PCI-E card for a customer who needs some functionality that
they bought 15 years ago on an EISA card. Most people will not be able to do
that themselves, so they are stuck: buy a new microscope, or pay the
microscope manufacturer for their help. You may balk at the prices they
quote you, but they (the column manufacturers) either have to embark on a
re-development, which can take a lot of man-hours, or they had the foresight
to stock some of the older computers and keep them and the software in
storage for decades in case someone needs a computer in the future. This is
not a cheap option for the manufacturers, so don't be surprised at the price
tag.

TEMs are driven by technology just as much as they are by user demand. That
means automation and capabilities that are unthinkable without the help of
computers. But computers are driven by a completely different set of demands
(Remember Moore's Law?). And that is what causes the problem, I think.

For the user, I think there is something they can do: First, keep the
computer that runs the microscope as lean as possible. Don't weigh it down
with applications like word processing or other applications. If the
computer runs the microscope adequately, then it will do so still in 10
years. General computing may have surpassed the computer and OS a long time
ago, but it will run the microscope just as well as it did the first day.
Use a second computer to run software that is not necessary for running the
microscope itself. Some manufacturers do that anyway.
And third, if you are really concerned about the microscope computer
breaking down: Buy a second, identical computer and put it in storage. If
the first computer breaks, transfer the cards and hard disk, and you should
be up again in no time. If down the road one of the cards breaks, you only
need to find a replacement, which is a lot easier than having the board and
software re-engineered for a different standard.

These are just my 2 cents.

Mike Bode
---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com






-----Original Message-----
X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com]
Sent: Wednesday, June 30, 2010 9:10 AM
To: mike.bode-at-resaltatech.com

I've been reading this thread with some interest, as in a previous
life, I was a computer consultant (Although I don't like admitting it
in public.)  There are ways to make older software run on newer
operating systems and computers, and I wonder if the marriage between
the EM software and the computer is more of an artificial one.

For example, I received a JEOL DSG unit to use with the 35 that was
installed where I was teaching, and despite the software being written
for windows 3.1, it is now working perfectly on windows XP.  The only
limitation that I've found with that system is that the hardware is an
old ISA board, but I'm sure that with the proper schematics (Which I
have) and time (Which I don't have) I could re-wire it for a more
modern PCI card and get similar results without too much hassle.

I don't see any reason why a modern EM, computer equipped with at
least a PCI card shouldn't be end-user upgradable in both hardware and
OS, as long as you still use the same card.

I used to see this a lot with hardware vendors in the computing world
where one vendor would create a new version of the software which they
claimed required you to use updated hardware- they were full of it.
The new software worked fine on old hardware, and likewise, the old
software worked fine on new hardware.

I think it would be worth an experiment by someone with a slightly
older computer controlled scope to try installing the software on
another machine with a newer OS.

--Justin.



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From: ben.micklem-at-pharm.ox.ac.uk
Date: Mon, 5 Jul 2010 10:46:30 -0500
Subject: [Microscopy] Re: UV bulb replacement for Leica AFS1

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

Again I agree with Mike.
Let's put everything in context: what we publish is a condensed report of what we did during the last 3-6-36 monthes (depending if you are married with children or single, in which case you can spend 16 hours a day in the lab). Obviously you cannot show everything you saw during the 100+ hours you spent on the TEM. So you have to (wisely if possible) choose one or a few pictures which represent what you have observed. You can see it as a set of data or as a picture, still it is incomplete (it's just a sample from lots of pictures) inaccurate (which technique, especially in EM, does show "the truth"? They all introduce artifacts, extractions and so on) and subjective (you choose it as "representative") even if you do your best. Knowing that, do you think that the displacement of a few pixels in a whole picture/set of data really matters? I don't think so.
If you show a nice colocalization in fluorescence microscopy, resampling the picture won't change the message.

Have a nice hot summer week-end (or chilly winter week-end for those in the southern hemisphere).

Stephane






 
----- Original Message ----
X-from: "mike.bode-at-resaltatech.com" {mike.bode-at-resaltatech.com}
To: nizets2-at-yahoo.com
Sent: Thu, July 1, 2010 7:21:26 PM

As someone who has been in the "imaging business" for a while, I'd like to
comment on this thread. I think there are at least 2 issues that are being
discussed here:

1) publishing images in journals
2) dealing with meta data

Regarding issue 1, I don't really see a big issue, but perhaps I am
mistaken. When I published, I never thought of the images in the publication
as anything more than visual data to prove a point, which would be described
in the text. I never thought that someone would look at the image in the
magazine at the pixel level and try to draw any conclusions from that. Given
that, a resampling of the image using a bi-cubic spline (or other) doesn't
really matter that much (provided no artificial artifacts like Moire are
introduced). It's even easier for online publications: provide a link to the
original data. Am I totally off the mark here? I mean, if you need to show
data at the pixel level, I would not rely on people using a magnifying glass
to see the data.

Dealing with meta data is a more complex issue. There are standards (TIF,
for example) that allow meta data to be stored with the image in a (sort of)
standardized way. The problem is that microscopy is a tiny little fraction
of the imaging world, and the standard bearers are not focused on microscopy
(no pun intended). For example, TIF has a tag for "Resolution". The problem
is that it is used differently from the way a microscopist would use it. It
is essentially the print resolution. So, if you specify that as, say, 10-6,
or a micron, a program such as Word will try to print it at that resolution.
A 1kx1k image would simply print out as 1mm x 1mm. Not what is intended.
There are really no standard microscopy tags in the TIF definition. That
leaves the other option: use private tags. That works fine as long as you
stay in the same software, but software from another manufacturer cannot
read the tags, so there is no interoperability.

With the microscopy community so small (relatively speaking), I think it is
up to us (perhaps MSA?) to define some internal standards and possibly try
to get them embedded in a general standard. Even if it the acceptance within
the TIF (or other) standard is a long shot, if MSA would publish a list of
keywords and tags to be used, I think that most software manufacturers would
quickly adopt it and at least provide interoperability among microscopy
software. Most manufacturers of cameras and software, for example, collect a
whole range of data (camera data, microscope data, imaging conditions, etc.)
which are then stored somehow (sometimes in private tiff tags, others use a
separate text file, etc.). the problem is getting everybody on the same
page.

Mike Bode
---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com








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From kefwyprawydiz-at-wyprawy.net Fri Jul 2 07:33:25 2010
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Well, I think this is a valid perspective, however I also think that if a $30-40K maintenance upgrade can be prevented by engineering an easy, economical way to do it into the instrument up front, that would be the way to go. When I think of the word "upgrade" I tend to think of additional accessories and capabilities being added on to the basic platform, rather than simply maintaining usability.

I mean, would you buy a new car and stock a couple extra engines for it as a hedge against not being able to drive it five years down the road because now everybody uses a different type of engine. (Not a great analogy, I admit, but you catch my drift.)

Not being an hardware or software engineer, I really don't know the feasibility of all this. Makes for interesting discussion, though.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Friday, July 02, 2010 5:40 AM
To: Tindall, Randy D.

I completely agree with that. I think it is a mistake to buy a 1 million $ worth machine and think that you won't need any additional expenses during the next 20 years. When you buy the microscope (and not 5-10 years later), it is wise to calculate the financing of the necessary updates.
When you have a budget for 1 Million $ (or half of it if you like), it seems reasonable to keep 30-40 k$ for future upgrades. Or as Mike said, just buy 3 identical computers and you have enough harware at your disposal. Preventing instead of curing has always been a winner strategy ;-)

Stephane

 


----- Original Message ----
X-from: "mike.bode-at-resaltatech.com" {mike.bode-at-resaltatech.com}
To: nizets2-at-yahoo.com
Sent: Thu, July 1, 2010 7:44:09 PM

Frankly, I don't think it is that easy. Computers and Operating systems
evolve, and it is not always possible to simply install software on a new
OS. If you are talking just about software, I think it might be possible,
although even there I have my doubts. Hardware is where the problem lies.
Let's simply take a look at the standards for add-on cards: ISA, EISA, PCI,
PCI-E. None of these is backward compatible, and you can't find any PCs with
the first 2 anymore. Even PCI is hard to find nowadays.
Now, remember, the cards that you find to control a microscope are most
likely custom made cards, and a microscope manufacturer will likely not
re-develop a PCI-E card for a customer who needs some functionality that
they bought 15 years ago on an EISA card. Most people will not be able to do
that themselves, so they are stuck: buy a new microscope, or pay the
microscope manufacturer for their help. You may balk at the prices they
quote you, but they (the column manufacturers) either have to embark on a
re-development, which can take a lot of man-hours, or they had the foresight
to stock some of the older computers and keep them and the software in
storage for decades in case someone needs a computer in the future. This is
not a cheap option for the manufacturers, so don't be surprised at the price
tag.

TEMs are driven by technology just as much as they are by user demand. That
means automation and capabilities that are unthinkable without the help of
computers. But computers are driven by a completely different set of demands
(Remember Moore's Law?). And that is what causes the problem, I think.

For the user, I think there is something they can do: First, keep the
computer that runs the microscope as lean as possible. Don't weigh it down
with applications like word processing or other applications. If the
computer runs the microscope adequately, then it will do so still in 10
years. General computing may have surpassed the computer and OS a long time
ago, but it will run the microscope just as well as it did the first day.
Use a second computer to run software that is not necessary for running the
microscope itself. Some manufacturers do that anyway.
And third, if you are really concerned about the microscope computer
breaking down: Buy a second, identical computer and put it in storage. If
the first computer breaks, transfer the cards and hard disk, and you should
be up again in no time. If down the road one of the cards breaks, you only
need to find a replacement, which is a lot easier than having the board and
software re-engineered for a different standard.

These are just my 2 cents.

Mike Bode
---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com






-----Original Message-----
X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com]
Sent: Wednesday, June 30, 2010 9:10 AM
To: mike.bode-at-resaltatech.com

I've been reading this thread with some interest, as in a previous
life, I was a computer consultant (Although I don't like admitting it
in public.)  There are ways to make older software run on newer
operating systems and computers, and I wonder if the marriage between
the EM software and the computer is more of an artificial one.

For example, I received a JEOL DSG unit to use with the 35 that was
installed where I was teaching, and despite the software being written
for windows 3.1, it is now working perfectly on windows XP.  The only
limitation that I've found with that system is that the hardware is an
old ISA board, but I'm sure that with the proper schematics (Which I
have) and time (Which I don't have) I could re-wire it for a more
modern PCI card and get similar results without too much hassle.

I don't see any reason why a modern EM, computer equipped with at
least a PCI card shouldn't be end-user upgradable in both hardware and
OS, as long as you still use the same card.

I used to see this a lot with hardware vendors in the computing world
where one vendor would create a new version of the software which they
claimed required you to use updated hardware- they were full of it.
The new software worked fine on old hardware, and likewise, the old
software worked fine on new hardware.

I think it would be worth an experiment by someone with a slightly
older computer controlled scope to try installing the software on
another machine with a newer OS.

--Justin.



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From international-at-uli.ca Sat Jul 3 01:16:08 2010
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Colleagues,
For the record, in reply to this questioner, I sent a reprint
of our work doing this with cucumber hypocotyls. Not a leaf but
perhaps of use. The citation is to Marga et al. 2005 Plant Journal
43: 181-190. I didn't want to send an attachment to the list. Anyone
wanting a reprint of this is welcome to ask me.
Tobias
}
}
} Email: tylerjhayden-at-gmail.com
} Name: Tyler Hayden
}
} Organization: University Lincoln-Nebraska
}
} Title-Subject: [Filtered] Microfibril preparation for SEM/ESEM
}
} Message: I'm looking for a method to prepare a leaf sample for the
} SEM to visualize the microfibril orientation. Any ideas or past
} experience on the subject?
}
} Tyler
}



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www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

==============================Original Headers==============================
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From news-at-bulletinnewspapers.com Sun Jul 4 22:25:55 2010
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I've just looked, and our last bulb was made by Cathodeon. Their website now
re-directs to {http://www.heraeus-noblelight.com/en/home.html} so maybe they
have been taken over. Looking through that site, you can get to this form,
that seems promising:

{http://www.heraeus-noblelight.com/en/uv-curing-exposure/products-for-uv-cur
ing-and-exposure/replacement-lamps.html}

Ben

--
Imaging Technician
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271866
{http://mrcanu.pharm.ox.ac.uk/}


On 01/07/2010 18:50, "marisa_ote-at-yahoo.com" {marisa_ote-at-yahoo.com} wrote:

}
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} Hello,
}
} I just heard from my local Leica sales representative that Leica does not
} provide customers with UV bulbs for the AFS1 anymore. The only available
} option is to buy a new attachment (a modified version of the UV LED attachment
} of the newer AFS2) for $2,200!!!! Does anyone know about other suppliers
} selling this type of UV bulb or a cheaper solution that does not involved
} spending $2,200? Thanks
}
} Marisa
}
} Marisa Otegui
} Department of Botany
} University of Wisconsin-Madison



==============================Original Headers==============================
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From: yu_zongsen-at-yahoo.com
Date: Mon, 5 Jul 2010 11:28:42 -0500
Subject: [Microscopy] EDS quantification in DM

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

Dear Colleagues,


We encountered a problem when we quantified EDS spectrum in Gatan Digital Micrograph. The problem is: we got a very different result when we use the K-line or L-line for EDS quantification. For instance, Nb-L line and Nb-K line, using Nb-K line for quantification gives a reasonable result while using Nb-L gives a wrong result. I am attaching two quantification results (using Nb-L and Nb-K line), respectively .


Does any one know what makes so large discrepancies ?



Material: Ti50 % -Al 45% -Nb 5%, nominal composition (at %), EDS spectra were acquired in TEM mode.

------------------------------------------------------------------
quantification Mode: Standardless
Correction Mode: Cliff Lorimer
Fit index (chi**2): 99.4
Number of iterations: 1
Total atoms: 15.1
Total weight: 100.0
Element Line KRatio Correction Overvoltage Weight Atoms %
Ti Ka 0.628±0.0069 1.0000 0.00 0.63±0.01 53.08±0.58
Al Ka 0.288±0.0040 1.0000 0.00 0.29±0.00 43.27±0.61
Nb Ka 0.084±0.0085 1.0000 0.00 0.08±0.01 3.65±0.37


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

Quantification Mode: Standardless
Correction Mode: Cliff Lorimer
Fit index (chi**2): 99.4
Number of iterations: 1
Total atoms: 5.1
Total weight: 100.0
Element Line KRatio Correction Overvoltage Weight Atoms %
Ti Ka 0.153±0.0017 1.0000 0.00 0.15±0.00 22.55±0.25 Al Ka 0.070±0.0010 1.0000 0.00 0.07±0.00 18.39±0.26
Nb La 0.777±0.0332 1.0000 0.00 0.78±0.03 59.06±2.52
--------------------------------------------------------------------





==============================Original Headers==============================
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15, 27 -- From: zongsen yu {yu_zongsen-at-yahoo.com}
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From: charlotte.goodman-at-jhsmh.org
Date: Tue, 6 Jul 2010 19:39:00 -0500
Subject: [Microscopy] viaWWW: LOOKING FOR A USED ULTRAMICROTOME

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

Dear Colleagues,


We encountered a problem when we quantified EDS spectrum in Gatan Digital Micrograph. The problem is: we got a very different result when we use the K-line or L-line for EDS quantification. For instance, Nb-L line and Nb-K line, using Nb-K line for quantification gives a reasonable result while using Nb-L gives a wrong result. I am attaching two quantification results (using Nb-L and Nb-K line), respectively .


Does any one know what makes so large discrepancies ?


Best regards,

Zaoli


-----------------------------------------------
Material: Ti50 % -Al 45% -Nb 5%, nominal composition (at %), EDS spectra were acquired in TEM mode.

------------------------------------------------------------------
quantification Mode: Standardless
Correction Mode: Cliff Lorimer
Fit index (chi**2): 99.4
Number of iterations: 1
Total atoms: 15.1
Total weight: 100.0
Element Line KRatio Correction Overvoltage Weight Atoms %
Ti Ka 0.628±0.0069 1.0000 0.00 0.63±0.01 53.08±0.58
Al Ka 0.288±0.0040 1.0000 0.00 0.29±0.00 43.27±0.61
Nb Ka 0.084±0.0085 1.0000 0.00 0.08±0.01 3.65±0.37


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

Quantification Mode: Standardless
Correction Mode: Cliff Lorimer
Fit index (chi**2): 99.4
Number of iterations: 1
Total atoms: 5.1
Total weight: 100.0
Element Line KRatio Correction Overvoltage Weight Atoms %
Ti Ka 0.153±0.0017 1.0000 0.00 0.15±0.00 22.55±0.25 Al Ka 0.070±0.0010 1.0000 0.00 0.07±0.00 18.39±0.26
Nb La 0.777±0.0332 1.0000 0.00 0.78±0.03 59.06±2.52
--------------------------------------------------------------------





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17, 27 -- From: zongsen yu {yu_zongsen-at-yahoo.com}
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From matt-at-sturgisboatworks.com Mon Jul 5 19:08:07 2010
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Message-ID: {01cb1ca7$82cde980$eae4af4f-at-matt}

Questions have recently been posted on this list server about the
characteristics and functioning of the ordinary self-biased electron
gun. If any Listers are not too confident about their understanding
of the functioning and use of the common self-biased electron gun I
might suggest reading two articles that were recently published in
"Microscopy Today". The first is an article I wrote that just
appeared in the May 2010 issue (pages 26 to 30) in which contains a
simplified description of the mechanism by which saturation is
produced. The second is an article written by Steve Chapman that
appeared in the May 2009 issue (pages 46 to 48) , in which he gives
detailed descriptions of several methods for accurately achieving
saturation.

--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

==============================Original Headers==============================
2, 15 -- From bigelow-at-umich.edu Mon Jul 5 22:11:57 2010
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2, 15 -- Authuser bigelow;
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2, 15 -- To: Microscopy Listserver {microscopy-at-microscopy.com}
2, 15 -- From: Wil Bigelow {bigelow-at-umich.edu}
2, 15 -- Subject: [Microscopy]Electron Gun Info
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From matt-at-rume.com Tue Jul 6 03:00:05 2010
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Hi all,

To add to what Wil has written, Fred Schamber wrote an article for
_Microscopy_Today_ in 1999 entitled "Emission Myths" discussing the
operation of thermionic electron guns. I converted the the article to
pdf and posted it (with permission):

http://www.mse.eng.ohio-state.edu/~COLIJN/emission_myths.pdf

Cheers,
Henk

At 7/5/2010 11:14 PM, bigelow-at-umich.edu wrote:
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} Questions have recently been posted on this list server about the
} characteristics and functioning of the ordinary self-biased electron
} gun. If any Listers are not too confident about their understanding
} of the functioning and use of the common self-biased electron gun I
} might suggest reading two articles that were recently published in
} "Microscopy Today". The first is an article I wrote that just
} appeared in the May 2010 issue (pages 26 to 30) in which contains a
} simplified description of the mechanism by which saturation is
} produced. The second is an article written by Steve Chapman that
} appeared in the May 2009 issue (pages 46 to 48) , in which he gives
} detailed descriptions of several methods for accurately achieving
} saturation.
}
}

--
Hendrik O. Colijn
www.ceof.ohio-state.edu
OSU Campus Electron Optics Facility colijn.1-at-osu.edu
040 Fontana Labs (614) 292-0674 (V)
116 W. 19th Ave. (614) 292-7523 (F)
Columbus, OH 43210

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

==============================Original Headers==============================
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Title-Subject: [Filtered] ULTRAMICROTOME

Message: LOOKING FOR A USED ULTRAMICROTOME IN EXCELLENT CONDITION. I
PREFER A LEICA UC6 ULTRAMICROTOME BUT WILL CONSIDER OTHER
ULTRAMICROTOMES. ALSO, WILL NEED NECESSARY ACCESSORIES.

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From: cross-at-tru.ca
Date: Wed, 7 Jul 2010 19:15:56 -0500
Subject: [Microscopy] viaWWW: Enivironmental SEMS: Lab6 filaments vs. Schottky, etc.

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


On Jul 6, 2010, at 5:35 AM, colijn.1-at-osu.edu wrote:

} Hi all,
}
} To add to what Wil has written, Fred Schamber wrote an article for
} _Microscopy_Today_ in 1999 entitled "Emission Myths" discussing the
} operation of thermionic electron guns. I converted the the article to
} pdf and posted it (with permission):
}
} http://www.mse.eng.ohio-state.edu/~COLIJN/emission_myths.pdf
}
} Cheers,
} Henk
}
} At 7/5/2010 11:14 PM, bigelow-at-umich.edu wrote:
} }
} } Questions have recently been posted on this list server about the
} } characteristics and functioning of the ordinary self-biased electron
} } gun. If any Listers are not too confident about their understanding
} } of the functioning and use of the common self-biased electron gun I
} } might suggest reading two articles that were recently published in
} } "Microscopy Today". The first is an article I wrote that just
} } appeared in the May 2010 issue (pages 26 to 30) in which contains a
} } simplified description of the mechanism by which saturation is
} } produced. The second is an article written by Steve Chapman that
} } appeared in the May 2009 issue (pages 46 to 48) , in which he gives
} } detailed descriptions of several methods for accurately achieving
} } saturation.
} }

Hi Henk,
Thanks for pointing out Fred's article. I spent 20+ years working on
a TEM that had an independent bias on the gun and a set of NiCd
batteries in the HT tank to provide the filament voltage, and I always
adjusted the voltage so that the W filament tip looked the smallest
and was uniform. At lower filament voltage, the tip did not look
uniform, which, I was taught, indicated that some parts of the tip did
not emit electrons that made it into the beam--presumably due to lower
temperature, higher work function, or both. In addition, we would
plug the tips into a jig that provided good electrical contact, but
not rigid mechanical contact and heated them briefly to white heat
(under vacuum, of course) before the tips were installed into Wehnelt
assemblies. This was done to remove any stresses that might have
built up during manufacture of the tip. (After a time, all W tips we
bought had been treated this way, so we no longer had to do this on
our own.) When operated under these conditions, the filaments lasted
several hundred hours, and often for 1000 hours or more.
When I worked on a TEM with a LaB6 tip, I was able to see several
regimes as the voltage was increased. At a relatively low filament
voltage, only the flat part of the truncated pyramid of the tip
emitted. The beam was pretty dim, but very coherent--good for
diffraction, which does not require high beam current. As the
filament voltage increased, four lobes appeared, which corresponded to
emission from the four sides of the pyramid, then the lobes coalesced
into a uniform small spot--the usual operating condition--as emission
from the edges of the pyramid also contributed to the beam. I was
told that these regimes were due to the differences in work function
for the different crystal orientations in the tip.
Although Fred says in his article that he does not like the term
"saturation" to describe these phenomena, If what I had been told is
correct, then the term is, indeed, appropriate, since it would refer
to uniform emission from all parts of the tip, at which point further
increase in filament voltage would not produce higher beam current.
Yours,
Bill


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From mitch.hagen-at-rhi.com Wed Jul 7 01:23:38 2010
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Email: drg.mitchell-at-sydney.edu.au
Name: David Mitchell

Organization: University of Sydney

Title-Subject: [Filtered] TEM: CCD counts into electrons

Message: Dear Listers

I have a Gatan Ultrascan camera (2k x 2k). I am using
DigitalMicrograph 1.82. I understand how to convert from counts in an
image to electrons (via the Target control panel). However, the
problem I have is that the calibration does not appear to have been
done at installation. I have a conversion efficiency of 6.262 - which
I presume is counts per electron. The problem is I do not know where
to plug in this number in order to enable the conversion from counts
to electrons. I have trawled through the tags in the Global Info
several times, but have not found anything likely. Any advice would
be greatly appreciated.

Thanks and regards, Dave Mitchell

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From jackiee-at-wiktel.com Wed Jul 7 08:21:40 2010
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Hi Folks.

I would appreciate any advice, suggestions, and words of caution concerning taking apart a Phillips EM410 TEM that will be disposed of or auctioned off for parts.

Cheers
Deb

Debra L. Moreau, Ph.D.
Microscopy & Image Analysis | Microscopie et analyse d'images
Food Safety & Quality | Salubrité et qualité des aliments
Agriculture and Agri-Food Canada | Agriculture et Agroalimentaire Canada
32 Main Street | 32 Rue Main
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Debra.Moreau-at-agr.gc.ca {mailto:Debbie.Moreau-at-agr.gc.ca}
Telephone | Téléphone 902-679-5710
Facsimile | Télécopieur 902-679-2311
Teletypewriter | Téléimprimeur 613-759-7470
Government of Canada | Gouvernement du Canada




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Email: cross-at-tru.ca
Name: Cynthia Ross Friedman

Organization: Thompson Rivers University

Title-Subject: [Filtered] Enivironmental SEMS: Lab6 filaments vs.
Schottky, etc.

Message: Hello - I am in the process of trying to choose an
environmental, high resolution (2nm) SEM (ideally for unprepared
biological specimens). Does anyone have any first-hand experience
with any "brands" and types of these? I suppose I cannot actively
ask for a suggestion on the ones I am considering, just because I
want to remain unbiased, but any comments -- positive and negative --
about any system you have used would be very helpful. If you can
pass on the names of end-users or other references, that would be
helpful as well.

If my posting is related to any thread that has been started before,
I apologize, as I am not really familiar with "list servs" and how
they operate, so a direction to any pre-existing threads would be
appreciated. I would be happy to take your comments and suggestions
outside of this forum, too - my e-mail is cross-at-tru.ca

Thanks for your help,
Cindy

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From: oshel1pe-at-cmich.edu
Date: Thu, 8 Jul 2010 08:30:34 -0500
Subject: [Microscopy] Re: Ask-A-Microscopist

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

I don't think that you are going to find an ESEM
with real resolution of 2nm. Others can correct me
of course.

As far as I see it, the high resolution figures are
at 25KV-30KV...hardly what conditions you seek. Unless
you coat the specimens, in which case, why have an ESEM?

IMO, highest resolution is going to be achieved at short
WD and with an in-lens detector. This seems to be counter
to what you are looking for. I would suggest that you
change 2nm to 20nm resolution. How much do you really need?

I think that the difference between thermionic sources and
Schottky sources is stability--thermal Schottky sources
are way more stable than thermionic ones, IMO. Perhaps you
can find some magic out there. I just don't see how you are
going to get 2nm resolution from any ESEM. It takes high
vac to achieve this level of resolution.

The difference between W and LaB6 is night and day. TFE
is at least ten times more bright than LaB6. But the contamination
and poisoning issue is serious for TFE sources, IMO.

If you are doing low mag imaging (you did not state your
needs in this regard), then LaB6 might work. But I do
not think that you are going to get TFE resolution at high
vacuum with a LaB6 ESEM.

My view on this. Choose wisely.

gary g.


At 05:18 PM 7/7/2010, you wrote:



} ----------------------------------------------------------------------------
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14, 21 -- From gary-at-gaugler.com Thu Jul 8 00:11:11 2010
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14, 21 -- Subject: Re: [Microscopy] viaWWW: Enivironmental SEMS: Lab6 filaments
14, 21 -- vs. Schottky, etc.
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From hameedsaedi-at-web2mail.com Thu Jul 8 00:47:29 2010
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Reply-To: {hameedalsaedi55-at-gmail.com}

Hi

I have to agree with Gary about performance and the environmental style SEM.
Whilst carrying out your survey be aware that they are not all called ESEM,
each manufacturer having its own special name for this type of imaging.

My only additional comment is that if you want resolution and
"natural/unprepared" specimens the solution is cryo which may be carried out
at high resolution. Perhaps you see cryo as being prepared but at least the
specimen is very near to its natural state.


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com]
Sent: 08 July 2010 06:13
To: protrain-at-emcourses.com

I don't think that you are going to find an ESEM
with real resolution of 2nm. Others can correct me
of course.

As far as I see it, the high resolution figures are
at 25KV-30KV...hardly what conditions you seek. Unless
you coat the specimens, in which case, why have an ESEM?

IMO, highest resolution is going to be achieved at short
WD and with an in-lens detector. This seems to be counter
to what you are looking for. I would suggest that you
change 2nm to 20nm resolution. How much do you really need?

I think that the difference between thermionic sources and
Schottky sources is stability--thermal Schottky sources
are way more stable than thermionic ones, IMO. Perhaps you
can find some magic out there. I just don't see how you are
going to get 2nm resolution from any ESEM. It takes high
vac to achieve this level of resolution.

The difference between W and LaB6 is night and day. TFE
is at least ten times more bright than LaB6. But the contamination
and poisoning issue is serious for TFE sources, IMO.

If you are doing low mag imaging (you did not state your
needs in this regard), then LaB6 might work. But I do
not think that you are going to get TFE resolution at high
vacuum with a LaB6 ESEM.

My view on this. Choose wisely.

gary g.


At 05:18 PM 7/7/2010, you wrote:



} ---------------------------------------------------------------------------
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14, 21 -- Subject: Re: [Microscopy] viaWWW: Enivironmental SEMS: Lab6
filaments
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==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member the
listserver, and any reply should go directly to the poster as well as
to the list.
(Not to me - Oshel)
==========================================================
} Below is the result of your form, submitted on Thursday, July 08,
} 2010 at 12:24:18 AM.
}
} realname - Beth Lampert
} Email - elisabethlampert-at-gmail.com
} ORGANIZATION - Oregon State University
} EDUCATION - Graduate College
} LOCATION - Oregon, USA
} SUBJECT_OF_QUESTION - Educational Programs in Microscopy
} QUESTION - Hi,
}
} I am curious to know if you know of any graduate level programs
} available for someone interested in biological microscopy, or any
} other educational resources to learn about microscopy. I am
} interested primarily in electron microscopy but haven't had too much
} experience with it and would love to learn more. Any advice would
} be much appreciated!
}
} Thanks!
} Beth


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From: kemmishtree-at-gmail.com
Date: Thu, 8 Jul 2010 09:03:04 -0500
Subject: [Microscopy] viaWWW: HD-2000 and/or HD-2300

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From: gantz-at-bu.edu
Date: Thu, 8 Jul 2010 11:05:00 -0500
Subject: [Microscopy] Darkroom Equipment Donations

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Dear Gary and All:

First, thank everyone so much for your rapid and helpful responses. This is an unbelievable forum and community!

I think I had better be a little clearer on my objectives, and I think Gary and others who replied to me privately are correct: no one SEM is "one size fits all".

In brief, I am a botanist interested in floral development and real-time floral events. As you can guess, I don't need 2 nm resolution to look at flowers! However, I want to make my SEM available to others besides myself, and I want it to be very easy to use. I am at a small institution, and will have a lot of students using the microscope. I need a SEM that is extremely user-friendly, and will require little specimen prep for the sake of the biology students. However, some of my colleagues are interested in nano-scale biological molecules, so I wanted to ensure they could use the microscope as well. I also want to ultimately couple to EDX for X-ray microanalysis. Hmmm...maybe I am asking for too much... However, I do see that I would probably have to ask those interested in the fine-scale work to prepare their samples.

I did not want to name names, and I hope this is OK, but I am now between two SEMS: the
Zeiss LS Evo 15 with Lab6 and the
Quanta 250 FEG (with Shottky emitter)

If any of you could comment on the suitability of either of these SEMs for the work I propose, or on the user-friendliness and cost-efficiency of either instruments, I would be thrilled. if you want to contact me off-list, my e-mail is: cross at tru.ca

Thanks again very much, and thanks for all of your help. I am so happy I posted here, and apologize for my obvious newbie status.

Cheers,
Cindy


**********************************
Cynthia Ross Friedman, Ph.D.
Associate Professor
Department of Biological Sciences
Thompson Rivers University
Box 3010, 900 McGill Road
Kamloops, British Columbia
V2C 5N3

Voicemail/phone: (250) 828-5424
Email: cross-at-tru.ca
Homepage: http://tru.ca/faculty/cross
**********************************


} } } Gary Gaugler {gary-at-gaugler.com} 7/7/2010 10:11 PM } } }
I don't think that you are going to find an ESEM
with real resolution of 2nm. Others can correct me
of course.

As far as I see it, the high resolution figures are
at 25KV-30KV...hardly what conditions you seek. Unless
you coat the specimens, in which case, why have an ESEM?

IMO, highest resolution is going to be achieved at short
WD and with an in-lens detector. This seems to be counter
to what you are looking for. I would suggest that you
change 2nm to 20nm resolution. How much do you really need?

I think that the difference between thermionic sources and
Schottky sources is stability--thermal Schottky sources
are way more stable than thermionic ones, IMO. Perhaps you
can find some magic out there. I just don't see how you are
going to get 2nm resolution from any ESEM. It takes high
vac to achieve this level of resolution.

The difference between W and LaB6 is night and day. TFE
is at least ten times more bright than LaB6. But the contamination
and poisoning issue is serious for TFE sources, IMO.

If you are doing low mag imaging (you did not state your
needs in this regard), then LaB6 might work. But I do
not think that you are going to get TFE resolution at high
vacuum with a LaB6 ESEM.

My view on this. Choose wisely.

gary g.


At 05:18 PM 7/7/2010, you wrote:



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From matt.wetherbee-at-alaskaair.com Thu Jul 8 10:53:47 2010
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Hello Everyone,
We are cleaning out our darkroom and would like to give away the
following equipment. We would prefer that a local/regional high school or
college pick up the equipment as shipping could be cumbersome.

1. Mohr Pro 8 Automatic Print Processor in good condition and in working
order.

2. Jobo CPE2 Plus Color Print Processor, Type 4065 which looks to be in good
shape but can't vouch for its functionality.

3. Arkay drum print washer, Model 1620A.

If you have interest please contact off-line.

Donald Gantz
Research Histologist/Electron Microscopist
Dept. Physiology and Biophysics
Center for Advanced Biomedical Research
Boston University School of Medicine
Email: gantz-at-bu.edu


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From: shafermr-at-whitman.edu
Date: Thu, 8 Jul 2010 13:35:54 -0500
Subject: [Microscopy] SEM - need help imaging bacteria

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

We recently purchased an FEI Quanta 250 SEM that is capable of ESEM. We are trying to image bacterial cultures (E. coli and B. cereus) and can't seem to see anything. We've looked at many papers to get starting parameters and tips, and still no luck. We've tried the following:

1) using the cold stage and ESEM mode, we've tried the following parameters: temp 4-5C, 300-600 Pa, RH 50-90%, 5-20 KV, spot size 4-5. We've tried a) smearing the culture on a coverslip and attaching it to the cold stage, b) adding the culture to filter paper (0.4 and 0.2 um), and c) directly dropping culture into the cold stage stub.

2) using low vac, smeared the culture onto a coverslip. Parameters were 10 KV, 130 Pa, and spot size of 4. We also tried adjusting all these parameters up and down and still no bacteria.

3) using high vac, smeared culture onto a coverslip and coated with Au. We also coated filter paper with sample on it. Parameters were 12.5 KV and spot size of 4. We tried adjusting these parameters up and down and still no bacteria.

Does anyone have any suggestions? Any input is greatly appreciated!! Ideally, we don't want to have to stain, fix, or wash the bacteria. Please feel free to email me directly at shafermr-at-whitman.edu

Thanks!
Michelle


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From: DusevichV-at-umkc.edu
Date: Thu, 8 Jul 2010 15:07:11 -0500
Subject: [Microscopy] RE: SEM - need help imaging bacteria

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Michelle,
Before going with ESEM, you need to get reliable results in high vac. It
looks like your problem is your culture medium. When you smear culture
on a substrate, after drying out you will have bacteria embedded in
dried salts, sugars, proteins. Since SEM is capable to see only a
surface of a specimen, bacteria will be blocked by a layer of dried
media components. The same is true for ESEM: we can see only surface. If
you specimen is submerged in water solution, you will see only surface
of solution. You need to dry out solution, so that bacteria will be
above it, but then again, it will be covered with dry components of
media. If your culture can survive for a while in distilled water, then
you have good chances to see it in the ESEM mode. Put a drop of
distilled water with bacteria on specimen stage (keep the stage far from
objective lens and EDS detector during initial pumping, otherwise
droplets from degassing water may damage or contaminate them). Changing
pressure or temperature let drop evaporate slowly, and you can observe
your specimen. May be for best results you will need to fix it. If the
culture cannot tolerate pure water, then you have to fix it.
Good luck,
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784


} -----Original Message-----
} From: shafermr-at-whitman.edu [mailto:shafermr-at-whitman.edu]
} Sent: Thursday, July 08, 2010 1:37 PM
} To: Dusevich, Vladimir
} Subject: [Microscopy] SEM - need help imaging bacteria
}
}
}
}
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}
} We recently purchased an FEI Quanta 250 SEM that is capable of ESEM.
We
} are trying to image bacterial cultures (E. coli and B. cereus) and
} can't seem to see anything. We've looked at many papers to get
starting
} parameters and tips, and still no luck. We've tried the following:
}
} 1) using the cold stage and ESEM mode, we've tried the following
} parameters: temp 4-5C, 300-600 Pa, RH 50-90%, 5-20 KV, spot size 4-5.
} We've tried a) smearing the culture on a coverslip and attaching it to
} the cold stage, b) adding the culture to filter paper (0.4 and 0.2
um),
} and c) directly dropping culture into the cold stage stub.
}
} 2) using low vac, smeared the culture onto a coverslip. Parameters
were
} 10 KV, 130 Pa, and spot size of 4. We also tried adjusting all these
} parameters up and down and still no bacteria.
}
} 3) using high vac, smeared culture onto a coverslip and coated with
Au.
} We also coated filter paper with sample on it. Parameters were 12.5 KV
} and spot size of 4. We tried adjusting these parameters up and down
and
} still no bacteria.
}
} Does anyone have any suggestions? Any input is greatly appreciated!!
} Ideally, we don't want to have to stain, fix, or wash the bacteria.
} Please feel free to email me directly at shafermr-at-whitman.edu
}
} Thanks!
} Michelle
}
}
} ==============================Original
} Headers==============================
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} 7, 24 -- Date: Thu, 8 Jul 2010 11:35:52 -0700 (PDT)
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5, 25 -- From DusevichV-at-umkc.edu Thu Jul 8 15:07:10 2010
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5, 25 -- From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu}
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From: wtivol-at-verizon.net
Date: Thu, 8 Jul 2010 15:38:54 -0500
Subject: [Microscopy] SEM - need help imaging bacteria

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

Dear Michelle & Vladimir,
If the bacteria cannot tolerate distilled water due to ionic strength
and/or pH, you could try NH4HCO3 or NH4Ac, which are volatile buffers
that evaporate along with the water, so there is no residue.
Yours,
Bill

On Jul 8, 2010, at 1:17 PM, DusevichV-at-umkc.edu wrote:

} Michelle,
} Before going with ESEM, you need to get reliable results in high
} vac. It
} looks like your problem is your culture medium. When you smear culture
} on a substrate, after drying out you will have bacteria embedded in
} dried salts, sugars, proteins. Since SEM is capable to see only a
} surface of a specimen, bacteria will be blocked by a layer of dried
} media components. The same is true for ESEM: we can see only
} surface. If
} you specimen is submerged in water solution, you will see only surface
} of solution. You need to dry out solution, so that bacteria will be
} above it, but then again, it will be covered with dry components of
} media. If your culture can survive for a while in distilled water,
} then
} you have good chances to see it in the ESEM mode. Put a drop of
} distilled water with bacteria on specimen stage (keep the stage far
} from
} objective lens and EDS detector during initial pumping, otherwise
} droplets from degassing water may damage or contaminate them).
} Changing
} pressure or temperature let drop evaporate slowly, and you can observe
} your specimen. May be for best results you will need to fix it. If the
} culture cannot tolerate pure water, then you have to fix it.
} Good luck,
} Vladimir
}
} Vladimir M. Dusevich, Ph.D.
} Electron Microscope Lab Manager
} 371 School of Dentistry
} 650 E. 25th Street
} Kansas City, MO 64108-2784
}
}
} } -----Original Message-----
} } From: shafermr-at-whitman.edu [mailto:shafermr-at-whitman.edu]
} } Sent: Thursday, July 08, 2010 1:37 PM
} } To: Dusevich, Vladimir
} } Subject: [Microscopy] SEM - need help imaging bacteria
} }
} }
} }
} }
} }
} -----------------------------------------------------------------------
} } -----
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} } America
} } To Subscribe/Unsubscribe --
} } http://www.microscopy.com/MicroscopyListserver
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} }
} } We recently purchased an FEI Quanta 250 SEM that is capable of ESEM.
} We
} } are trying to image bacterial cultures (E. coli and B. cereus) and
} } can't seem to see anything. We've looked at many papers to get
} starting
} } parameters and tips, and still no luck. We've tried the following:
} }
} } 1) using the cold stage and ESEM mode, we've tried the following
} } parameters: temp 4-5C, 300-600 Pa, RH 50-90%, 5-20 KV, spot size 4-5.
} } We've tried a) smearing the culture on a coverslip and attaching it
} } to
} } the cold stage, b) adding the culture to filter paper (0.4 and 0.2
} um),
} } and c) directly dropping culture into the cold stage stub.
} }
} } 2) using low vac, smeared the culture onto a coverslip. Parameters
} were
} } 10 KV, 130 Pa, and spot size of 4. We also tried adjusting all these
} } parameters up and down and still no bacteria.
} }
} } 3) using high vac, smeared culture onto a coverslip and coated with
} Au.
} } We also coated filter paper with sample on it. Parameters were 12.5
} } KV
} } and spot size of 4. We tried adjusting these parameters up and down
} and
} } still no bacteria.
} }
} } Does anyone have any suggestions? Any input is greatly appreciated!!
} } Ideally, we don't want to have to stain, fix, or wash the bacteria.
} } Please feel free to email me directly at shafermr-at-whitman.edu
} }
} } Thanks!
} } Michelle




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From: LettJ-at-ent.wustl.edu
Date: Thu, 8 Jul 2010 15:39:50 -0500
Subject: [Microscopy] TEM & SEM - Phosphate Buffer versus PBS

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

Can Sodium Phosphate Buffer be used interchangeably with
Phosphate-Buffered Saline for TEM and SEM processing purposes? (And
does it matter whether or not the PBS contains potassium?) We typically
use 0.1M Sodium Phosphate Buffer at pH 7.4.

Thank you,

Jaci

Jaclynn Lett
Senior Research Technician, EM Facility
Research Center for Auditory and Vestibular Studies Department of
Otolaryngology Washington University School of Medicine 660 S Euclid
Ave., Campus Box 8115 St. Louis, MO 63110

Office: 314-747-7257
Fax: 314-747-7230
Email: lettj-at-ent.wustl.edu
Website: http://otocore.wustl.edu



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From: ehaller-at-health.usf.edu
Date: Mon, 12 Jul 2010 10:16:18 -0500
Subject: [Microscopy] Philips EM300 Vacuum & Sorvall MT2B

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

The short answer is no. For one thing they are significantly different osmolarity (~200 vs 300 mOsm roughly). The higher phosphate buffer will also lead to more severe precipitation of calcium which looks ugly in the TEM. The lack of potassium found in physiological buffers is probably unimportant for fixation buffers.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: LettJ-at-ent.wustl.edu [mailto:LettJ-at-ent.wustl.edu]
Sent: Thursday, July 08, 2010 3:40 PM
To: Phillips, Thomas E.

Can Sodium Phosphate Buffer be used interchangeably with
Phosphate-Buffered Saline for TEM and SEM processing purposes? (And
does it matter whether or not the PBS contains potassium?) We typically
use 0.1M Sodium Phosphate Buffer at pH 7.4.

Thank you,

Jaci

Jaclynn Lett
Senior Research Technician, EM Facility
Research Center for Auditory and Vestibular Studies Department of
Otolaryngology Washington University School of Medicine 660 S Euclid
Ave., Campus Box 8115 St. Louis, MO 63110

Office: 314-747-7257
Fax: 314-747-7230
Email: lettj-at-ent.wustl.edu
Website: http://otocore.wustl.edu



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From jade6705-at-gmail.com Thu Jul 8 18:35:56 2010
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You are invited to submit an abstract to the "Microanalysis in
Geoscience: Advances and Challenges" session at the AGU Winter
Meeting Dec 13-17 meeting in San Francisco.

V29: Microanalysis in Geoscience: Advances and Challenges
Description: Advances in microanalytic technology and
instrumentation have given geoscientists new possibilities in
examining earth materials. Low kV electron probe microanalysis with
field emission guns, femto-second lasers, more versatile CL
detectors, high precision Ar/Ar mass spectrometry for geochronology,
EBSD-detectors hooked to SEMs, and improved SIMS analysis of smaller
volumes, to name some. Challenges exist, e.g. interpreting and
quantifying the results. There is an increasing need for development
of well characterized standards. Sample preparation may be a
stumbling block (e.g. EBSD). We particularly invite abstracts which
document unresolved problems. Co-sponsor: Microbeam Analysis Society

The deadline for abstracts is Sept 2 (online site for submissions
opens July 21).

Conveners
John Fournelle, University of Wisconsin, johnf-at-geology.wisc.edu
Brian Jicha, University of Wisconsin, bjicha-at-geology.wisc.edu
Heather Lowers, USGS, hlowers-at-usgs.gov
Alan Koenig, USGS, akoenig-at-usgs.gov

Please contact any of the conveners for more information. Thank you.

==============================Original Headers==============================
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From jackie-at-marjacq.com Fri Jul 9 16:53:07 2010
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Dear Colleagues,

I am seeking some advice on two issues:

EM300:
My Philips EM300 is experiencing a vacuum problem. At initial startup
the water valves open and the diffusion pumps heat up - the PVL and
HV indicator lamps are illuminated along with the Vacuum "ON" light -
when I press the "ON" button the PVL, PV1 and HV indicators are lit
and the forepump starts. After about 30 seconds a valve opens and the
rotary pump can be heard displacing air (the PV meter drops but then
returns to 100). The vacuum "ON" light goes dark after about 1-2
minutes.

On the meter, HV setting remains at 0 and the PV remains at 100. The
rotary pump just runs on without any other valves opening.

For about a year I had noticed that the scope pumped down to
operational vacuum in about 10 minutes - much faster than I thought
possible (main valve open and the HV light dark - the HV meter was
just above zero and in the past it was barely able to get to 20 - I'm
wondering if the meter is actually functioning). The filament
energized typically and did not oxidize so I assumed high vacuum was
achieved? Could the Penning tube be dirty (if so is there a typical
way to clean it)?

Any suggestions? Also, can anyone recommend a good third party
service engineer if I cannot resolve the problem - I am located on
Long Island, NY.

Sorvall Porter-Blum MT-2B:
I have a couple of these older ultramicrotomes that are experiencing
drive belt and position/duration belt issues (cracked and broken
belts). Does anyone know if these are simply o-rings? Does anyone
know of a replacement source? They seem easy enough to replace.

Thanks in advance for your assistance!

Steve
--
Stephen J. Beck
Professor
Bio-Imaging Center/Electron Microscopy
Department of Biology
Nassau Community College
Garden City, NY 11530
Voice Mail: (516) 572-7829
Email: {becks-at-sunynassau.edu}
URL: {http://www3.ncc.edu/faculty/BIO/becks/BECKS.HTM}

==============================Original Headers==============================
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9, 21 -- Subject: Philips EM300 Vacuum & Sorvall MT2B
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From internationalx-at-chester.ac.uk Sat Jul 10 14:03:29 2010
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Dear Colleagues,

We will be offering a Sunday short course at the Microscopy & Microanalysis
meeting 2010 in Portland OR. The course, although assigned to the Biology
section, could also be very beneficial to the Material Science community as it
was, when we taught a similar topic at the "Euro-summer school on Electron
Crystallography and Cryo-Electron Microscopy on Inorganic Materials and Organic
and Biological Molecules" held in 2001 in Barcelona.

Course description:

X12 3D Electron Microscopy of Macromolecular Assemblies
Teresa Ruiz, Michael Radermacher, and Stefan Birmanns.

This short course will provide a comprehensive description of the methods used
for 3D structure determination of macromolecular complexes from electron
micrographs. First specimen preparation techniques for single particles (deep
stain, vitreous ice), will be presented and the selection of imaging conditions
including low-dose imaging. This will be followed by a detailed explanation of
image processing techniques with special emphasis on the random conical
reconstruction technique. In the last part structure interpretation and docking
of X-ray structures to 3D EM densities will be demonstrated.

--

Michael Radermacher, Ph.D., Prof.
University of Vermont
Dept. Molecular Physiology and Biophysics
HSRF 120 / 149 Beaumont Ave
Burlington, VT 05405
USA

Tel: + 802 656-4834
Fax: + 802 656-0747


==============================Original Headers==============================
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From inmotiza-at-inmotiza.com Sun Jul 11 20:29:26 2010
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Hi, Stephen,

There's a gentleman named Bill McGee in Liverpool, New York that carries belts for Sorval microtomes. It's easy to replace the drive belts. You need basic tools, Super Glue Gel, a razor blade and the instructions that Bill will provide. The belt kits were $35 last year, part#16802 for an MT-2. Bill's phone# is 315-452-0828. Microtome Service Co., 7568 Florian Way, Liverpool, NY 13088. I have no financial ties to the company, just some old MT-2's :)

Ed Haller

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-cas.usf.edu
________________________________________
X-from: Stephen.Beck-at-ncc.edu [Stephen.Beck-at-ncc.edu]
Sent: Saturday, July 10, 2010 1:25 PM
To: Haller, Edward

Dear Colleagues,

I am seeking some advice on two issues:

EM300:
My Philips EM300 is experiencing a vacuum problem. At initial startup
the water valves open and the diffusion pumps heat up - the PVL and
HV indicator lamps are illuminated along with the Vacuum "ON" light -
when I press the "ON" button the PVL, PV1 and HV indicators are lit
and the forepump starts. After about 30 seconds a valve opens and the
rotary pump can be heard displacing air (the PV meter drops but then
returns to 100). The vacuum "ON" light goes dark after about 1-2
minutes.

On the meter, HV setting remains at 0 and the PV remains at 100. The
rotary pump just runs on without any other valves opening.

For about a year I had noticed that the scope pumped down to
operational vacuum in about 10 minutes - much faster than I thought
possible (main valve open and the HV light dark - the HV meter was
just above zero and in the past it was barely able to get to 20 - I'm
wondering if the meter is actually functioning). The filament
energized typically and did not oxidize so I assumed high vacuum was
achieved? Could the Penning tube be dirty (if so is there a typical
way to clean it)?

Any suggestions? Also, can anyone recommend a good third party
service engineer if I cannot resolve the problem - I am located on
Long Island, NY.

Sorvall Porter-Blum MT-2B:
I have a couple of these older ultramicrotomes that are experiencing
drive belt and position/duration belt issues (cracked and broken
belts). Does anyone know if these are simply o-rings? Does anyone
know of a replacement source? They seem easy enough to replace.

Thanks in advance for your assistance!

Steve
--
Stephen J. Beck
Professor
Bio-Imaging Center/Electron Microscopy
Department of Biology
Nassau Community College
Garden City, NY 11530
Voice Mail: (516) 572-7829
Email: {becks-at-sunynassau.edu}
URL: {http://www3.ncc.edu/faculty/BIO/becks/BECKS.HTM}

==============================Original Headers==============================
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From: spurgeon-at-drexel.edu
Date: Mon, 12 Jul 2010 17:46:15 -0500
Subject: [Microscopy] TEM - Sample suggestions for two-day outreach course

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

Hi

What's your favorite vendor for recoating fluorescent viewing screen for a
TEM? Thanks.

--
Pang (Wai Pang Chan, wpchan-at-u.washington.edu, PAB A087, 206-685-1519)
The Biology Imaging Facility (http://depts.washington.edu/if/)

==============================Original Headers==============================
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From jacquelynda.cook-at-stereotaxis.com Mon Jul 12 16:35:44 2010
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Hello everyone,

My university's materials science department will be hosting a two-day
short course designed to introduce undergraduate freshmen to
materials. I will be organizing a session on TEM and I am looking for
suggestions as to what samples to show the students. Since they are
just entering college and probably have never used a TEM, I think it
would be best to use a sample that is visually striking and will
capture their attention. The goal isn't really to teach microscopy,
but rather to motivate the students to pursue a career in materials.

I am considering some carbon nanotube samples but I am open to
alternatives, such as metals or ceramics. I would prefer samples that
are easy to prepare or ones that can be purchased online at a
reasonable price. Any suggestions would be much appreciated. Thank
you.

--
Steven Spurgeon
Dynamic Characterization Group
Department of Materials Science & Engineering
Drexel University

Office: Bossone 102
Email: spurgeon-at-drexel.edu
Web: http://www.materials.drexel.edu/dcg/


==============================Original Headers==============================
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From: wtivol-at-verizon.net
Date: Mon, 12 Jul 2010 19:01:52 -0500
Subject: [Microscopy] Re: TEM - Sample suggestions for two-day outreach course

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


On Jul 12, 2010, at 3:58 PM, spurgeon-at-drexel.edu wrote:

} Hello everyone,
}
} My university's materials science department will be hosting a two-day
} short course designed to introduce undergraduate freshmen to
} materials. I will be organizing a session on TEM and I am looking for
} suggestions as to what samples to show the students. Since they are
} just entering college and probably have never used a TEM, I think it
} would be best to use a sample that is visually striking and will
} capture their attention. The goal isn't really to teach microscopy,
} but rather to motivate the students to pursue a career in materials.
}
} I am considering some carbon nanotube samples but I am open to
} alternatives, such as metals or ceramics. I would prefer samples that
} are easy to prepare or ones that can be purchased online at a
} reasonable price. Any suggestions would be much appreciated. Thank
} you.
}
} --
} Steven Spurgeon
} Dynamic Characterization Group
} Department of Materials Science & Engineering
} Drexel University
}
} Office: Bossone 102
} Email: spurgeon-at-drexel.edu
} Web: http://www.materials.drexel.edu/dcg/
}
Dear Steven,
I posted this suggestion last month.
Yours,
Bill

} I am not a materials person, but I did have a project looking at
} sediment from river bottoms. The samples are very easy to prepare--
} just suspend the mud in a fairly dilute suspension, let the larger
} particles settle, then put a drop (a few ul will do) onto a carbon/
} formvar coated grid and let dry. If my experience was typical, you
} will see diatom skeletons, mineral fragments, crystalline and not,
} and other items in a TEM image. EDS (if available) will also show
} some surprises for some of the particles. I saw U and Au in
} addition to the usual suspects, Na, K, Si, Al, Ti, Ca, etc. Good
} luck.

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From: susan.trant-at-viha.ca
Date: Mon, 12 Jul 2010 19:47:54 -0500
Subject: [Microscopy] viaWWW: Decontaminating Diamond Knives

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Name: Sue Trant

Organization: Vancouver Island Health Authority

Title-Subject: [Filtered] Decontaminating Diamond Knives

Message: Hello all

I was fortunate enough to aquire two new diamond knives. There is a
decontamination certificate to send the old scratched knives back to
Leica. Is there some sort of protocol to decontaminate/clean diamond
knives? There does not appear to be a protocol on the web. An email
to Leica has not be answered as yet.

Thank you in advance

Sue Trant
EM Technologist
Department of Pathology
Vancouver Island Health Authority

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From: Frank_Karl-at-lincolnelectric.com
Date: Tue, 13 Jul 2010 06:17:11 -0500
Subject: [Microscopy] Re: TEM - Sample suggestions for two-day outreach course

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Email: connie-at-ultrapathimaging.com
Name: Connie Cummings

Organization: UltraPath Imaging Clinic Inc

Title-Subject: [Filtered] Zeiss SEM Model DSM960

Message: Hi all!
I am writing today to ask if anyone out there has a need for a Zeiss
SEM Model DSM960 for parts only. If interested please get in touch
with me and I'll let you know more information.
Thanks,
Connie Cummings
email:connie-at-ultrapathimaging.com

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From ghj-at-sdfs.com Tue Jul 13 02:57:59 2010
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I would suggest carbon black, say an high structure HAF. The prep is
simple. A small, no make that a tiny, amount of carbon black
ultrasonicated in ChCl3 (you just need a ultrasonic cleaning) and place a
micro drop on a carbon film grid. CB has a grape cluster-like appearance.
Morphology of CB has a direct relationship to it's reinforcing properties
in rubber. The original air filled tubed tires were, I've read, compounded
with Mg oxide and lasted 3-400 miles! Now, thanks largely to CB you feel
robbed if you don't get 26-30,000 miles with high speed driving and wet
traction!!

Consider precipitated silica and kaolin clay as well. The preps the same
but each morphology is unique and easy to recognize.

I once burnt a small ribbon of Mg and caught some of the fumes on a carbon
coated grid. The morphology was very amazing. Again less is more at this
level of sample prep so don't encrust your grid.

Let us know what you chose and how it works out.

Frank Karl



spurgeon-at-drexel.e
du
To
07/12/2010 06:55 frank_karl-at-lincolnelectric.com
PM cc

Subject
Please respond to [Microscopy] TEM - Sample
spurgeon-at-drexel.e suggestions for two-day outreach
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Hello everyone,

My university's materials science department will be hosting a two-day
short course designed to introduce undergraduate freshmen to
materials. I will be organizing a session on TEM and I am looking for
suggestions as to what samples to show the students. Since they are
just entering college and probably have never used a TEM, I think it
would be best to use a sample that is visually striking and will
capture their attention. The goal isn't really to teach microscopy,
but rather to motivate the students to pursue a career in materials.

I am considering some carbon nanotube samples but I am open to
alternatives, such as metals or ceramics. I would prefer samples that
are easy to prepare or ones that can be purchased online at a
reasonable price. Any suggestions would be much appreciated. Thank
you.

--
Steven Spurgeon
Dynamic Characterization Group
Department of Materials Science & Engineering
Drexel University

Office: Bossone 102
Email: spurgeon-at-drexel.edu
Web: http://www.materials.drexel.edu/dcg/


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From: swalck-at-southbaytech.com
Date: Tue, 13 Jul 2010 12:36:51 -0500
Subject: [Microscopy] TEM - Sample suggestions for two-day outreach course

Contents Retrieved from Microscopy Listserver Archives
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Sue,

Unless you have used the knives for cryo sectioning, why should they be decontaminated? If sectioning resin embedded tissues, there should not be a problem with infectious agents, unless you have sectioned prions, then long term high temperatures are needed. If you truly need to decontaminate the knife, soaking in 3% glut for a week should be acceptable.

Ed


Edward P. Calomeni
Director EM Lab - Pathology
The Ohio State University
M018 Starling Loving Hall
320 W. 10th Ave.
Columbus, OH 43210
614-293-5580 (office)
614-293-8806 (lab)
edward.calomeni-at-osumc.edu
-----Original Message-----
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Email: susan.trant-at-viha.ca
Name: Sue Trant

Organization: Vancouver Island Health Authority

Title-Subject: [Filtered] Decontaminating Diamond Knives

Message: Hello all

I was fortunate enough to aquire two new diamond knives. There is a decontamination certificate to send the old scratched knives back to Leica. Is there some sort of protocol to decontaminate/clean diamond knives? There does not appear to be a protocol on the web. An email to Leica has not be answered as yet.

Thank you in advance

Sue Trant
EM Technologist
Department of Pathology
Vancouver Island Health Authority

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From jacobsen-at-sarcos.com Tue Jul 13 07:34:05 2010
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You can prepare thin sections of MoS2 single crystals very easily. MoS2 is
a dry lubricant that works because of its crystal structure. You can buy
flakes of the material. What I did was to take a good size flake and wax it
down on a glass slide with low temperature wax. (SBT Quickstick 135 would
work). Then take Scotch® tape and repeatedly press it onto the flake and
remove layers. Examine the sample with transmitted light through the glass
until the sample is thin. Completely dissolve the wax in acetone and
collect the sample on holey carbon grid. You get a neat structure of
dislocation networks in a BF image.

-Scott
 
Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA  92673
 
US Toll Free: 1-800-728-2233
Tel: (949) 492-2600
Fax: (949) 492-1499
 
www.southbaytech.com
swalck-at-southbaytech.com


-----Original Message-----
X-from: spurgeon-at-drexel.edu [mailto:spurgeon-at-drexel.edu]
Sent: Monday, July 12, 2010 3:57 PM
To: swalck-at-southbaytech.com

Hello everyone,

My university's materials science department will be hosting a two-day
short course designed to introduce undergraduate freshmen to
materials. I will be organizing a session on TEM and I am looking for
suggestions as to what samples to show the students. Since they are
just entering college and probably have never used a TEM, I think it
would be best to use a sample that is visually striking and will
capture their attention. The goal isn't really to teach microscopy,
but rather to motivate the students to pursue a career in materials.

I am considering some carbon nanotube samples but I am open to
alternatives, such as metals or ceramics. I would prefer samples that
are easy to prepare or ones that can be purchased online at a
reasonable price. Any suggestions would be much appreciated. Thank
you.

--
Steven Spurgeon
Dynamic Characterization Group
Department of Materials Science & Engineering
Drexel University

Office: Bossone 102
Email: spurgeon-at-drexel.edu
Web: http://www.materials.drexel.edu/dcg/


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From: z.zhou-at-sheffield.ac.uk
Date: Tue, 13 Jul 2010 20:11:06 -0500
Subject: [Microscopy] viaWWW: electro-polishing Ni-based superalloy

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The following request is being posted on behalf of an individual who is looking for microscopy images for an upcoming project he is considering. Please reply to him directly if you wish to participate.

******************************************
A Call for Participation
Greetings electron microscopists! Scott Whittaker has kindly offered to forward this message to you. My name is Michael Benson, and among other projects over the last few years I have been writing and image-processing for a series of art books for Abrams, the leading publisher of illustrated books in the US. Until now they have focused on space themes (the first, 'Beyond: Visions of the Interplanetary Probes,' was a 320 page volume of solar system landscape photography; the second, 'Far Out: A Space Time Chronicle,' essentially treated the rest of the universe - meaning nebulae, star clusters, galaxies, and galaxy clusters, etc). For more on my work please check out my website, www.kinetikonpictures.com.
The reason I'm writing you is that my next book proposes to investigate micro-worlds, and in fact will be called 'Nanocosmos.' And I have always been fascinated by the weird compelling beauty of what you are doing with the SEM. So the purpose of this mail is to announce the project and invite your participation in whichever form might be feasible. I am looking for compelling images, of course, be they of animal plant or mineral (though the first two will likely predominate). I am also looking for larger file sizes, as the book will be 11 x 11," with many spreads being 11 x 22" (across two pages). Also, this is an art book, and so reproduction quality will be extremely high, which of course benefit image sizes of, say, just under 2,000 pixels on the horizontal axis or more (with 2,576, to name one dimension I saw in Scott's lab last week, very nice indeed). So I'm seeking stunning images of large file size.
But just as much as large file sizes, I am very interested in finding large quantities of images to go through - hundreds of thousands, if possible. This is because it is rare to find that specific set of characteristics that comprise a truly superb image that can be understood as superb even by people outside of science (people in the arts, etc). I am very willing to do the work to plow through great quantities of images - in fact, I enjoy it thoroughly.
I hasten to add, I am also a science writer, and apart from interesting imagery I will be researching and writing about what is being investigated, and why.

What do you get out of it? Well, this is (I would submit) a superb outreach opportunity, and should you elect to participate of course your institutions and you yourselves by name will be handsomely acknowledged. (Publishing, like journalism is shrinking, and my advance is barely enough to keep me in coffee - so there is no money in this, just glory.!) I think what you are doing with the SEM is very little-known, even less understood, and this is a real opportunity to get some attention to the field. For one, you don't have a single, PR-savvy organization such as NASA behind you, but are more atomized among various research centers. (I also hope to have an exhibit of prints in the end, but I'm getting way ahead of myself.)
So, I very much hope you will contact me. My e-mail address is kinetikonpictures-at-yahoo.com. I am leaving for a relatively long summer trip to Europe today, and won't be back until August 9th, but hope you will write me anyway - I will certainly respond in August. I very much hope you will consider participation, which doesn't have to involve much of your time at all - I am simply seeking access to pictures. I should add that I will be happy to sign non-disclosure agreements that specify that when I have isolated the images I am truly interested in using, I need to contact the researchers who have made the images directly to ask for permission to use them, and the like. We can work out the modalities! The important thing is to get the project going -- and ultimately, shine a light on the superb work you have been doing. And that's what Nanocosmos will do.
Thanks a lot for your time and attention (and consideration).
All best,
Michael Benson


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


Scott Whittaker
Head NMNH Imaging
Manager SEM Lab
Smithsonian Institution
National Museum of Natural History
PO Box 37012   MRC104
Washington DC 20013-7012
202-633-0891




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Email: z.zhou-at-sheffield.ac.uk
Name: Zhou

Organization: The University of Sheffield

Title-Subject: [Filtered] electro-polishing Ni-based superalloy

Message: Dear Folks,

Can I ask for some advice on TEM sample prep using jet electro-polishing?

Sampe is Ni based super alloy with a content of
gamma prime. We were electro polishing in 5%
perchloric acid in methanol, using a Struers
Tenu-pol, - 40ƒC, flow rate of 10 or 8 and a
voltage of 30 or 35 V.

We are struggling to get a large enough
transparent area for any useful TEM. We are
either getting many small holes or 1 massive
hole... very little transparent area.

Any advice is highly appreciated, such as which
parameter we should change to make it better.

Zhou


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From: gary-at-perfendo.com
Date: Tue, 13 Jul 2010 20:11:40 -0500
Subject: [Microscopy] viaWWW:Looking for Used Philips 201 electron microscope

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Email: gary-at-perfendo.com
Name: Gary B. Carr

Organization: Pacific Endodontic Research Foundation

Title-Subject: [Filtered] Used Philips 201 electron microscope

Message: I just had the electronic box and HT tank and rotary pump
stolen from my research foundationfor my Philps 201. I am looking to
purchase either an intact Philips 201 or just those items that were
stolen. Anyone having an old Philips 201 can find a good home for it
here!.

Pacific Endodontic Research Foundation
6235 Lusk Blve
San Diego, CA 92121

858-558-3636
Dr. Gary B. Carr


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From: drg.mitchell-at-sydney.edu.au
Date: Tue, 13 Jul 2010 22:14:46 -0500
Subject: [Microscopy] viaWWW: Re: TEM: electro-polishing Ni-based superalloy

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Email: drg.mitchell-at-sydney.edu.au
Name: David Mitchell

Organization: University of Sydney

Title-Subject: [Filtered] TEM: electro-polishing Ni-based superalloy

Message: Dear Zhou

The recipe you are using should give you good results. I have used it
on pure nickel and austenitic stainless steels of many different
kinds with considerable success. However, a couple of things you can
play with are listed below:

You mention using a voltage of 30 to 35V. Many electropolishing
recipes suggest a particular voltage, when in fact they should
specify a current. It is current which will control the rate of
electrochemical dissolution of your specimen rather than voltage. In
fact the voltage will vary with the age of your solution. As you
polish more specimens the metal ion concentration will increase and
the resistance of the solution will change (generally fall).
Therefore, if you are keeping voltage constant during your
preparation, you will be using higher currents as you continue
polishing.

The current you to use should be around 120 - 180mA for double sided
thinning. Generally I start at around 120mA and then for subsequent
specimens I increase the current in steps of 20mA until I hit a sweet
spot. I then stick at that current and during electropolishing
endeavour to keep the current constant by varying the voltage.

If you use too little current, you will effectively anodise your
specimen and it will be covered in oxide and be generally pretty
poor. If you use too much current it will perforate very rapidly -
you won't be able to control it, and you'll end up with a specimen
which looks like an aperture - large hole, no thin areas.

Nickel is ferromagnetic, and your superalloy may be too. If so,
you'll want to make your blank foils as thin as possible. I generally
aim for around 100um for non-ferromagnetics but grind to 60-80um for
ferromagnetics. This helps minimise abberations in the microscope.
You do need good metallographic practice to make sure you do not
inject a lot of mechanical damage into such thin specimens and
obviously handle with care to avoid bending. These foils should be
perforating after perhaps 20-40s of electropolishing. If this is
occurring in much shorter than this then you may be using too much
current.

In terms of flow and stop sensitivity. I always leave the termination
sensitivity on maximum. Flow is of second order importance. On a
Tenupol 3 I set the flow rate to about 1/3rd full scale.

A couple of other points - as soon as the termination alarm sounds,
remove the specimen and plunge it into a beaker of methanol and then
wash in fresh methanol as quickly as possible. Thin areas of foil
will corrode very quickly. Finally, single phase materials in the
annealed condition generally electropolish beautifully. Introduce
second phases or cold work and you can get local perforations around
the second phases or pitting along dislocations. Your foil can end up
looking like an Australian road sign in the outback (peppered with
bullet holes). You may have to settle for what you can get.

Finally, even when you get a really good recipe, what works today,
may not work so well tomorrow. Electropolishing is one of the more
perplexing mystic arts.

Regards

Dave Mitchell

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From: oshel1pe-at-cmich.edu
Date: Wed, 14 Jul 2010 15:21:02 -0500
Subject: [Microscopy] Tri-Iodide/KOH etch

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==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member the
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(Not to me - Oshel)
==========================================================
realname - Carter Nakashima
Email - carter.nakashima-at-maxim-ic.com
SUBJECT_OF_QUESTION - Tri-iodide vs KOH
QUESTION - What is the difference between Tri-iodide and KOH when
used to etch/decorate integrated circuit layers? They both attack Al
but is there a difference in selectivity in other materials? Does
Tri-iodide attack oxides?



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From: spurgeon-at-drexel.edu
Date: Wed, 14 Jul 2010 15:44:20 -0500
Subject: [Microscopy] TEM - Postdoctoral Position Available in Drexel University's

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Postdoctoral Position Available Immediately: In Situ TEM

Dynamic Characterization Group - Department of Materials Science &
Engineering, Drexel University

The Dynamic Characterization Group (http://dcg.materials.drexel.edu/),
in the Department of Materials Sciences and Engineering at Drexel
University, is seeking a Ph.D. scientist with expertise in
transmission electron microscopy (TEM) and a background in either
materials science or condensed matter physics for a postdoctoral
position to perform in situ TEM experiments on electronic, structural,
and catalytic materials (most notably, ferroelectric/ multiferroic
ceramics, ultra high strength steels and next generation nuclear
reactor materials).

The candidate is required to be experienced in microscopy techniques
such as HRTEM, STEM and STEM-EDS, and FIB. Experience in
instrumentation development, in situ TEM, and EELS are highly
desirable. Applications will be accepted until the position is filled.

Anticipated start-date: September 2010

Please send a cover letter, curriculum vita, and names of three
references to: Dr. Mitra Taheri (mtaheri-at-coe.drexel.edu)

--
Steven Spurgeon
Dynamic Characterization Group
Department of Materials Science & Engineering
Drexel University

Cell: 719.330.0441
Office: Bossone 102
Email: spurgeon-at-drexel.edu
Web: http://www.materials.drexel.edu/dcg/

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From: Jerzy.Gazda-at-ceriumlabs.com
Date: Wed, 14 Jul 2010 16:59:56 -0500
Subject: [Microscopy] Tri-Iodide/KOH etch

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-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Wednesday, July 14, 2010 3:30 PM
To: Gazda, Jerzy

==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member the
listserver, and any reply should go directly to the poster as well as
to the list.
(Not to me - Oshel)
==========================================================
realname - Carter Nakashima
Email - carter.nakashima-at-maxim-ic.com
SUBJECT_OF_QUESTION - Tri-iodide vs KOH
QUESTION - What is the difference between Tri-iodide and KOH when
used to etch/decorate integrated circuit layers? They both attack Al
but is there a difference in selectivity in other materials? Does
Tri-iodide attack oxides?



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From: gary-at-gaugler.com
Date: Wed, 14 Jul 2010 17:12:54 -0500
Subject: [Microscopy] Re: Tri-Iodide/KOH etch

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I am not familiar with these chemicals. I use 50:1
BOE for decorating polished cross sections. It needs
to be at a consistent and stable temperature. Some
experimentation is needed for each type of die based
on metal and ILD.

For planar etch, I use plasma with SF6, Cl and O2 with
inclusion and exclusion, ratios and time based on the makeup
of the die. More experimentation.

BOE eats Al quickly but not so fast at all for Cu.

What are you trying to do/see? It seems to me that Maxim
is mostly a linear house. In this case, why not just use
LM DIC?

gary g.


At 01:22 PM 7/14/2010, you wrote:



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From: anmiv-at-drexel.edu
Date: Wed, 14 Jul 2010 19:51:52 -0500
Subject: [Microscopy] viaWWW: Sample heating under effect of electron beam in SEM

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Name: Anmiv S Prabhu

Organization: Drexel University

Title-Subject: [Filtered] Sample heating under effect of electron beam in SEM

Message: Hello all,

I was wondering if anyone here has carried out a systametic study of
the how samples heat up under the effect of the electron beam in an
SEM? And given the beam parameters (diameter, accelerating voltage,
scan rate etc.) and the properties of the sample (geometry, thermal
conductivity, geometry etc)is it possible to predict/calculate the
rise in temperature of the samlpe?

I thank you in advance.

Regards
Anmiv S Prabhu
PhD Candidate
School of Biomedical Science and Engineering
Drexel University
Philadelphia

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From: mlibbee-at-gmail.com
Date: Wed, 14 Jul 2010 19:52:20 -0500
Subject: [Microscopy] viaWWW: Intro to Focused Ion Beams

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From: ALawrence-at-entomology.msstate.edu
Date: Mon, 19 Jul 2010 05:55:40 -0500
Subject: [Microscopy] M&M 2010 note

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Email: tiancu-at-tx.technion.ac.il
Name: THEODORE IANCU

Organization: Rappaport Faculty of Medicine

Title-Subject: [Filtered] Breakdown TEM

Message: To: microscopy-at-microscopy.com


The meeting is just two weeks away and we still need more student
bursaries or volunteers to do things such as provide support in the
different symposia, monitor use of the Internet Café, or staff the
volunteer office.

Student bursaries are paid $10/hour for their assistance (paid by check
at the end of the meetings) with an added bonus of $10 cash for each
morning and/or afternoon session worked to help with meals Volunteers
are also eligible for the $10 cash for meals.

We can*t do it without help from you, so please consider serving as a
student bursary or volunteer. If anyone has any questions about the
bursary/volunteer program, or would like to participate, please
contact:

Amanda Lawrence
Electron Microscope Center
Mississippi State University
662-325-3019
alawrence-at-entomology.msstate.edu



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From: kpruessner-at-gmail.com
Date: Mon, 19 Jul 2010 07:34:26 -0500
Subject: [Microscopy] Position available: Director of Microscopy and Microanalysis Unit,

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Director - Microscopy and Microanalysis Unit (MMU)

The University is one of the major research and teaching institutions
in South Africa, with an international reputation in many fields. The
University aspires to move into the Top 100 ranking of World
Universities and to this end has in recent years invested heavily in
infrastructural development, including the acquisition of five new
electron microscopes. The instruments currently located in the MMU are
transmission and scanning electron microscopes, scanning probe and
confocal microscopes. The latest microscopes are a FEI Tecnai G2
Spirit TEM, FEI Quanta 400F Environmental SEM and a FEI Nova 600
NanoLab FIB SEM.

The University is seeking a Director for the Unit, to replace the
retiring incumbent, to take up the post not later than 3 January 2011,
but preferably sooner than that.

The Director of the Unit will operate a University facility which
services users in Life Sciences, Geology, Chemistry, Physics,
Materials, Metallurgy and Engineering, coming from both inside and
outside of the University. His or her duties will include the day to
day running of the Unit, the maintenance and service arrangements for
the instruments, the management of a staff of six, future planning for
replacement instruments and the infrastructure to house them and
budgetary responsibilities. He or she will periodically serve on
appropriate University committees. The Director is encouraged to
remain research active, to conduct his or her own research and to
collaborate with other researchers on microscopy-based projects.

The Director, with the support of his or her staff, will be expected
to train research students and staff on the how to use the
instruments. Other duties include facility management, technician
supervision and training in analysis of collected data. Demonstrations
for undergraduates should be given where appropriate. The Director
must be able to lecture on electron microscopy. He or she is expected
to be active in local and international Microscopy Societies and to
attend and participate in appropriate conferences, both locally and
abroad, in order to stay abreast with developments in technology and
instrumentation.

Experience with sample preparation and electron microscopy
investigations of biological and physical science materials is
essential, together with interpersonal skills for interaction with a
broad user base. Significant experience using scanning and
transmission electron microscopes for research is required.  A Ph.D.
in an appropriate discipline (Physics, Chemistry, Materials Science,
Chemical Engineering, Polymer Science, or one of the Life Sciences) is
essential. Prior experience of managing a similar facility would be a
strong recommendation.

A job description may be obtained from Iain.Burns-at-wits.ac.za, who can
also answer non-technical questions about the post and the Unit.

Questions of a scientific or technical nature should be directed to any one of:

Lesley.Cornish-at-wits.ac.za  (materials science and chemical engineering)
Karin.Pruessner-at-wits.ac.za (physics)
Paul.Franklyn-at-wits.ac.za (chemistry)
David.Mycock-at-wits.ac.za (life sciences)

Applications should include a full curriculum vitae, including a
publications list, the names and electronic contact details of four
referees and a supporting motivation written by the candidate. This is
an academic post with a professorial salary package, which includes
medical aid and pension.

Applications may be lodged in electronic or hard copy format and
should be sent to Yule.Banda-at-wits.ac.za, or to Mr Y Banda, Head:
Central HR Office, University of the Witwatersrand, Johannesburg,
Private Bag 3, PO Wits 2050, South Africa.

The closing date for applications is 27 August 2010.


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13, 35 -- Subject: Position available: Director of Microscopy and Microanalysis Unit,
13, 35 -- University of the Witwatersrand
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From: j.janssen-at-nki.nl
Date: Mon, 19 Jul 2010 07:40:57 -0500
Subject: [Microscopy] viaWWW: job opening in Amsterdam

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Email: j.janssen-at-nki.nl
Name: Hans Janssen

Organization: the Netherlands Cancer Institute

Title-Subject: [Filtered] job opening in Amsterdam

Message: A post-doctoral position and a PhD candidate position are
available in the group of Prof. Peter J. Peters, Department of Cell
Biology, Netherlands Cancer Institute, Amsterdam.
{http://www.nki.nl/research/peters}

The candidate will join an on-going project, which has the ultimate
goal to image biological nano-machines and their mode of action
within cells at macromolecular-scale by pushing the current limits of
nanotechnologies.

As cryo electron tomography is approaching macromolecular resolution,
creating maps of individual macromolecular complexes within cells is
becoming real. X-ray and NMR data can be 'docked' or fitted into the
lower resolution particle density maps to create a macromolecular
atlas of the cell under normal and pathological conditions. The
majority of cells, however, are too thick to be imaged in an intact
state and therefore we have built nano-chambers of 200 nm thickness
in which mammalian cells voluntarily migrate before they are
vitrified.

In particular, we now aim:

(1) To apply and further improve the technologies developed in the
course of the project so far for correlative cryo-LM.
(2) To use and develop methods for visualising vitreous biological
samples for cryo-TEM
(3) To use cryo-electron tomography to examine the sub-cellular
distribution and organisation of macromolecular structures involved
in cellar migration, phagocytosis of mycobacteria or CTL / target
cell interactions, depending on personal interest.
Cell. 2007 Jun 29;129(7):1287 , J Struct Biol. 2010 Feb;169(2):2

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10, 11 -- From: j.janssen-at-nki.nl (by way of MicroscopyListserver)
10, 11 -- Subject: viaWWW: job opening in Amsterdam
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From: Lucy.Collinson-at-cancer.org.uk
Date: Mon, 19 Jul 2010 08:22:47 -0500
Subject: [Microscopy] Job Opportunity: Scientific Officer Electron Microscopy

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

Scientific Officer
Electron Microscopy Core Technology Facility
London Research Institute
Holborn, London

Our London Research Institute is looking for an enthusiastic Scientific
Officer to work on biological electron microscopy projects in collaboration
with multiple research groups within the institute.

About the Role
As the Scientific Officer in the Electron Microscopy Unit you will take on a
range of projects using both transmission and scanning electron microscopy
in collaboration with multiple research groups within the institute. Under
the guidance of the lab head you will also research and implement new
imaging techniques and technologies. You will aid in day-to-day running of
the unit including training scientists in electron microscopy, optimisation
of protocols and maintenance of equipment.

About You
You will have a degree and preferably a PhD in biological sciences. Previous
knowledge of cell biology and light microscopy is essential. Experience of
use and routine maintenance of transmission and scanning electron
microscopes, basic specimen preparation techniques for electron microscopy
and digital image acquisition/ analysis are highly desirable.

About Cancer Research UK
The Cancer Research UK London Research Institute (LRI) is part of the
largest independent research organisation in the world and is committed to
training the next generation of cancer research scientists. The Electron
Microscopy Unit is a Core Technology Facility providing state-of-the-art
high resolution imaging for the research scientists at the London Research
Institute.

How to apply
All applications should be made via Cancer Research UK¹s careers website.
http://jobs.cancerresearchuk.org/view_vacancy.php?requirementId=10167&work_s
phere=Scientific
Closing date: 27th July 2010

--
Lucy Collinson PhD
Head of Electron Microscopy
London Research Institute
Cancer Research UK
44 Lincoln's Inn Fields
London
WC2A 3PX

Tel: 020 7269 3416
Fax: 020 7269 2805


This communication is from Cancer Research UK. Our website is at www.cancerresearchuk.org. We are a charity registered under number 1089464 and a company limited by guarantee registered in England & Wales under number 4325234. Our registered address is 61 Lincoln's Inn Fields, London WC2A 3PX. Our central telephone number is 020 7242 0200.

This communication and any attachments contain information which is confidential and may also be privileged. It is for the exclusive use of the intended recipient(s). If you are not the intended recipient(s) please note that any form of disclosure, distribution, copying or use of this communication or the information in it or in any attachments is strictly prohibited and may be unlawful. If you have received this communication in error, please notify the sender and delete the email and destroy any copies of it.

E-mail communications cannot be guaranteed to be secure or error free, as information could be intercepted, corrupted, amended, lost, destroyed, arrive late or incomplete, or contain viruses. We do not accept liability for any such matters or their consequences. Anyone who communicates with us by e-mail is taken to accept the risks in doing so.


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10, 30 -- Date: Mon, 19 Jul 2010 14:21:57 +0100
10, 30 -- Subject: Job Opportunity: Scientific Officer Electron Microscopy
10, 30 -- From: Lucy Collinson {Lucy.Collinson-at-cancer.org.uk}
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From: paul.gerroir-at-xrcc.xeroxlabs.com
Date: Mon, 19 Jul 2010 08:42:01 -0500
Subject: [Microscopy] LM - Illuminators

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

July 19, 2010

Hello All,
When using our PLM in reflected light mode, unwanted reflections from a relatively featureless surface mar image quality. To remedy this situation I have a fibre optic system with gooseneck illuminators but it is rather bulky and difficult to position the light guides in close where the gap between the objective and sample is small. Does anyone have a suggestion to overcome this problem? Are there now available smaller, more compact illuminating systems better suited for this application?

Thanks,
Paul

Paul J. Gerroir

Microscopy
Materials Characterization
Xerox Research Centre of Canada
2660 Speakman Drive
Mississauga, Ontario L5K 2L1

Phone: 905-823-7091, ext.216
FAX: 905-822-7022
e-mail: paul.gerroir-at-xerox.com




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From: qizhang-at-mail.ucf.edu
Date: Mon, 19 Jul 2010 11:26:56 -0500
Subject: [Microscopy] Post Doctoral Researcher opening at Penn State Univ

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

JOB TITLE: Post Doctoral Researcher
LOCATION: Penn State Materials Research Institute, University Park PA 16802

POSITION SUMMARY

Perform transmission electron microscopy to aid in the development of new electronic materials and metallurgical alloys. Work closely with researchers with expertise in semiconductor device processing and computational materials science.

SPECIFIC DUTIES

1. Perform TEM and scanning TEM (STEM) characterization of electronic and structural materials, with special emphasis on the interfaces between metals and semiconductors and the grain boundaries of metal alloys.

2. Prepare TEM specimens via FIB and possibly electropolishing and/or mechanical thinning and ion milling.

3. Perform phase identification via electron diffraction.

4. Perform compositional analysis using EDS and possibly EELS.

5. Interact (along with PIs) with industrial and government sponsors through conference calls and program review presentations.

TO APPLY: Please send a cover letter and CV by e-mail to Prof. Suzanne Mohney (mohney-at-ems.psu.edu).




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13, 17 -- From: "Qi Zhang" {qizhang-at-mail.ucf.edu}
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13, 17 -- Subject: Post Doctoral Researcher opening at Penn State Univ
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From: dave-at-boeckeler.com
Date: Mon, 19 Jul 2010 17:09:57 -0500
Subject: [Microscopy] High Pressure Freezer User Group Meeting

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


Dear Microscopists,

If you are planning to attend the forthcoming M&M 2010
meeting in Portland,
and have an interest in High Pressure Freezing,
please consider attending the following user group
meeting:

High Pressure Freezer User Group Meeting

Monday, August 2, 2010
5:30pm until around 7:30pm
Doubletree Hotel Portland, Mt. St. Helens Room
1000 NE Multnomah
Portland, OR 97232
(just a short walk from the Convention Center)

Meeting Philosophy: An informal, relaxed, discussion
group, sharing
experiences with high pressure freezing of biological
samples. Although
this meeting is primarily focused on experiences with
the HPM-010 High
Pressure Freezing Machine, it is open to anyone with
an interest in
cryoimmobilization by high pressure freezing or with
experiences to
share using other freezing devices.

Meeting Co-Chairs: Andres Kaech, PhD
Center for Microscopy and Image
Analysis
University of Zurich

Frank Macakuso
Director, Analytical Imaging
Facility
Albert Einstein College of
Medicine, Bronx, NY
Meeting Agenda:

5:30pm Welcome & Introduction:

5:30pm - 6:00pm 'A brief history of the
HPM-010 High Pressure Freezing
Machine and the birth of high
pressure freezing in Zurich'.
Andres Kaech, PhD

6:00pm - 6:30pm 'Preparative methods for
C.elegans ultrastructure and
immunoEM'
David Hall, PhD
Center for C. elegans Anatomy
Dominick P. Purpura Department
of Neuroscience
Albert Einstein College of
Medicine, Bronx, NY

6:30pm - 7:30pm Open discussion

Complimentary soft drinks, light refreshments, coffee
and tea will be served plus a cash bar will be open
during the meeting serving wine, beer and cocktails.

Please drop me an e-mail if you would like to reserve
a place.

See you in Portland

Dave Roberts
Director-RMC Products
Boeckeler Instruments Inc
4650 S. Butterfield Drive
Tucson, Arizona 85714
Tel: 520-745-0001
Fax: 520-745-0004
www.rmcproducts.com
dave-at-boeckeler.com

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From: doc.vrdoljak-at-gmail.com
Date: Mon, 19 Jul 2010 18:20:18 -0500
Subject: [Microscopy] fluorescent imaging stains compatible with Alkanes

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

Hello,
I've never worked extensively with alkanes, but does anyone know of
fluorescent stains I could use for imaging that are compatible,
soluble, and stable in 100% C10-C12 alkanes such as decane or
dodecane?

A quick search in google suggests something like Nile Red or DFSB-C0
Clear Blue Fluorescent Dye. This will be used to mix with the alkane
and monitor penetration of the alkane through a porous material by
fluorescence.
thanks.

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From: bumajdad-at-kuc01.kuniv.edu.kw
Date: Tue, 20 Jul 2010 09:20:13 -0500
Subject: [Microscopy] viaWWW: Job Vacancy Kuwait University

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This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at
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Email: bumajdad-at-kuc01.kuniv.edu.kw
Name: Ali Bumajdad

Organization: Kuwait University

Title-Subject: [Filtered] Job Vacancy

Message: Dear Colleague, I would like to bring your attention to the
following job available at the Electron Microscopy Unit at Kuwait
University. The unit is a well established one since more than 35
years.


Position vailable: Chief Technician for Electron Microscopy Unit,
Faculty of Science

Kuwait University
The Faculty of Science at Kuwait University is seeking to appoint a
chief technician position for Electron Microscopy Unit (EMU). The
applicant should hold an academic qualification of a good first
degree in material science or related subject preferably with a
master or doctorate degree. This must be accompanied with at least 5
years hand-on experience on Electron Microscopy operational
techniques but applicants with more than 10 years of experience in
practical microscopy or research are also recommended. Applicants
with substantial knowledge in crystallographic image processing of
electron diffraction micrographs and pattern analyses are also
appreciable. Applicants with experience of using SPM/AFM/STM for
sample analysis are appreciable. Successful candidates must be
capable of supervising a team of staff technicians and capable of
conducting training and research activities. Applicants also must
have good spoken and written English communication skills with
scientific publications.
The position is expected to be filled by september 2010. Salary is
commensurate with skills and experience.

Interested individuals should send a letter of interest, a CV, and
the contact information of three references to the Director of
Electron Microscopy Unit (EMU), Faculty of Science, University of
Kuwait (P.O. Box 5969 Safat 13060, e mail: emu-at-sci.ku.edu.kw)

For Further details please visit our website http://emu.kuniv.edu


Please also include a letter that refer to Advertisement in Kuwait
Times dated 17 July.

This is a good permanent position in faculty of science. Salary could
be in the neighbourhood of KD 1,200/- (1KD about 3.2 US dollars). 2
months paid leave. We are looking for someone with Ph.D and at least
5 + years experience in HRTEM and AFM (Material science)




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17, 11 -- From: bumajdad-at-kuc01.kuniv.edu.kw (by way of MicroscopyListserver)
17, 11 -- Subject: viaWWW: Job Vacancy Kuwait University
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==============================End of - Headers==============================




From: Lucy.Collinson-at-cancer.org.uk
Date: Tue, 20 Jul 2010 10:06:00 -0500
Subject: [Microscopy] Job Opportunity: Scientific Officer - Super Resolution LM

Contents Retrieved from Microscopy Listserver Archives
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Scientific Officer/Senior Scientific Officer
Super Resolution Microscopy Facility
Cancer Research UK London Research Institute
Holborn, London


Overview

We are looking for a Scientific officer/Senior Scientific Officer to join
our Super Resolution Microscopy Facility at the London Research Institute.

Details of the role
As part of the establishment of a new Super Resolution Microscopy Facility
at the London Research Institute we are recruiting a Scientific Officer /
Senior Scientific Officer to a key new role. We wish to recruit an
enthusiastic and very dedicated person who will take a prominent role in
setting up, running and further developing this emerging area of imaging.

The primary focus of the Super Resolution Microscopy Facility is to
facilitate the development and broad usage of super-resolution imaging and
data analysis to research laboratories within the LRI. The team will provide
state of the art support and advice regarding the design and execution of
experiments involving super-resolution imaging. The post holder will also
develop image analysis protocols for general implementation at the LRI.

About you
You will take a leading role in the day to day running of super resolution
and other high performance microscopes in consultation with the Head of LRI
Research Facilities and group leaders involved in high-resolution microscopy
and high content analysis. A good understanding of the latest applications
and technologies pertaining to advanced microscopy is essential. Previous
knowledge of super resolution microscopy and instrumentation would be
desirable.

About Cancer Research UK
The Cancer Research UK London Research Institute (LRI) is part of the
largest independent research organisation in the world and is committed to
training the next generation of cancer research scientists. The LRI conducts
innovative basic biological research to improve our understanding of cancer
and has made significant breakthroughs since its inception, and continues to
be at the forefront of its field. With more than 550 staff and students, the
LRI also has a strong academic programme and is committed to training the
next generation of world-leading scientists.

How to Apply
All applications should be made via Cancer Research UK¹s careers website.
For more information and to apply online please visit
http://jobs.cancerresearchuk.org/view_vacancy.php?requirementId=10189&work_s
phere=Scientific

Ref 10189

Closing date: 3rd August 2010

---
Lucy Collinson PhD
Head of Electron Microscopy
London Research Institute
Cancer Research UK
44 Lincoln's Inn Fields
London
WC2A 3PX

Tel: 020 7269 3416
Fax: 020 7269 2805


This communication is from Cancer Research UK. Our website is at www.cancerresearchuk.org. We are a charity registered under number 1089464 and a company limited by guarantee registered in England & Wales under number 4325234. Our registered address is 61 Lincoln's Inn Fields, London WC2A 3PX. Our central telephone number is 020 7242 0200.

This communication and any attachments contain information which is confidential and may also be privileged. It is for the exclusive use of the intended recipient(s). If you are not the intended recipient(s) please note that any form of disclosure, distribution, copying or use of this communication or the information in it or in any attachments is strictly prohibited and may be unlawful. If you have received this communication in error, please notify the sender and delete the email and destroy any copies of it.

E-mail communications cannot be guaranteed to be secure or error free, as information could be intercepted, corrupted, amended, lost, destroyed, arrive late or incomplete, or contain viruses. We do not accept liability for any such matters or their consequences. Anyone who communicates with us by e-mail is taken to accept the risks in doing so.


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9, 30 -- Subject: Job Opportunity: Scientific Officer - Super Resolution LM
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From: reinhard.rachel-at-biologie.uni-regensburg.de
Date: Tue, 20 Jul 2010 14:27:32 -0500
Subject: [Microscopy] Sterility of organisms in Epoxy

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Although this issue was raised early in June, I still think it is ok to post my opinion, my reply - which was left "unsent" in the folder "in progress" ...

} Message: Has anyone had dealings with the USDA, CDC, or HHS, or other
} agencies with respect to proving sterility of agents that have been
} fixed (4%paraformaldehyde/1%gluteraldehyde), osmicated(1% osmium),
} (potentially irradiated 2X16[6] rads), dehydrated, and embedded in
} epoxy.
}
} I find myself getting questions about things that never were
} questioned in the past.

as a microbiologist and EM specialist in Germany, I am lucky that nobody raised this question to me, so far (no USDA, CDC, HHS, ...).
I would bet that no (or hardly any?) microorganism = cell will survive the usual preparation / embedding protocol (usual: at RT). But who knows?!? what about viruses, spores? extremophiles? I am working with those extremophiles since 20 years now, none of which was so far shown to be infectious to human beings. In all studies, we were and are surprised to learn what they do tolerate.
For me, the worst step in EM preparation is not necessarily, or not only, the FA/GA and Os fixation/oxidation (do these substances fully penetrate and react with those molecules which are "important" for infectivity? in viruses, in spores, in vegetative cells?), but in particular the stepwise dehydration at RT in Ethanol or Acetone, and the final infiltration of the (hydrophobic, monomeric) resin. During these steps, most (all?) cells are very likely (!?!) to be destroyed, as the hydrophobic part of the membranes, the lipids, are extracted to great extent, and the membrane is not intact any more. During the final polymerization, almost all (?) bio-molecules are altered, inevitably.
But, there are question marks; and, what happens to the cells during freeze substitution? during embedding in LR white or gold, or Lowicryls? we electron microscopists are happy about the good, much improved structural preservation. Full stop. Does this - vice versa - mean that (few of) these cells are in a status to survive? at least their DNA? some mRNA? how to test this? And what about those few cells which were not fully embedded in the resin, due to "infiltration problems", and fall out of the section during the sectioning process? is this "cell" / cell debris / protein / ?? potentially infectious, or of harm to the operator or experimentator? How to test this, who wants to test this?

I do not want to make the sitution complicated. Understanding of all these processes is necessary, awareness is needed. - For me, personally, we have to protect ourselves from all the dangerous chemicals (FA, GA, Os, resin, resin dust, etc.). There is the real hazard potential, everyday.

And, yes, my bet is that not a single cell does survive, in particular the "routine embedding at room temperature". - But, which microorganism, which officer, takes care of my bet?

kind regards,
Reinhard


--
PD Dr. Reinhard Rachel
Universitaet Regensburg
Centre for EM / Anatomy
Faculty for Biology&Preclin. Med.
Universitaetsstr. 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29



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From: swalck-at-southbaytech.com
Date: Tue, 20 Jul 2010 17:32:42 -0500
Subject: [Microscopy] first published paper on ion milling

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Does anybody know what the first published paper was that used ion milling
to prepare a TEM sample? If you have it, is it possible that I can get the
reference and if possible a copy of the paper?

-Scott
 
Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA  92673
 
US Toll Free: 1-800-728-2233
Tel: (949) 492-2600
Fax: (949) 492-1499
 
www.southbaytech.com
swalck-at-southbaytech.com






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From: gradice-at-richmond.edu
Date: Tue, 20 Jul 2010 20:26:31 -0500
Subject: [Microscopy] viaWWW: Director of Microscopy and Imaging -University of Richmond

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This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: gradice-at-richmond.edu
Name: Gary Radice

Organization: University of Richmond

Title-Subject: [Filtered] Position available, Richmond, VA

Message: Director of Microscopy and Imaging

University of Richmond, Department of Biology

The University of Richmond invites applications for a Director of
Microscopy and Imaging, a continuing appointment that is not eligible
for tenure but includes faculty status. Applicants should have an MS
or PhD degree, expertise with light and electron microscopy, facility
with digital imaging, and the equivalent of two years experience
managing microscope facilities. The candidate must meet these
criteria by the time of selection for the position.

This position requires strong organizational and technical skills, an
ability to work with a diverse body of scientists, and an interest in
participating in a variety of research projects in the sciences.
Duties including include operation and routine maintenance of a TEM,
SEM, and laser scanning confocal microscope and associated equipment,
and other departmental microscopes. The Director will train and
supervise faculty and student users, assist with specimen preparation
and imaging for light and electron microscopy, and provide advice on
analysis of results. The Director will also work with faculty to
develop and maintain appropriate classroom activities and equipment
for undergraduates.

Applicants should apply online at https://www.urjobs.org using the
Faculty (Instructional/Research) link. Applicants must submit
electronically a letter of application and a curriculum vita. Final
candidates will be asked to submit three letters of reference. Review
of applications will begin on October 1, 2010 and continue until the
position is filled.

The University of Richmond is a highly selective private university
with approximately 3000 undergraduates located on a beautiful campus
six miles west of the heart of Richmond and in close proximity to the
ocean, mountains, and Washington, D.C. The University of Richmond is
committed to developing a diverse workforce and student body and to
supporting an inclusive campus community. We strongly encourage
applications from candidates who will contribute to these goals. For
more information, please see http://biology.richmond.edu or contact
Dr. Gary Radice (gradice-at-richmond.edu), Department of Biology, 28
Westhampton Way, University of Richmond, VA, 23015.


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From: admin-at-protochips.com
Date: Tue, 20 Jul 2010 20:26:59 -0500
Subject: [Microscopy] viaWWW: Job Opening @ Protochips

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Email: admin-at-protochips.com
Name: Kristen McLamb

Organization: Protochips

Title-Subject: [Filtered] Job Opening -at- Protochips

Message: Description:
Using technical expertise with customer facing skills to join the
Protochips' Applications Specialist team. In this role you will
travel up to 50% to perform customer demonstrations of Protochips'
electron microscopy equipment, give technical presentations, and
compose applications notes and sales support material in the field of
in situ electron microscopy.

For more information, please see:
http://protochips.com/en/about-us/positions.html

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From: oshel1pe-at-cmich.edu
Date: Wed, 21 Jul 2010 15:06:59 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist

Contents Retrieved from Microscopy Listserver Archives
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==========================================================
Forwarded from "Ask a Microscopist"
Please remember that the original poster is likely not a member the
listserver, and **any reply should go directly to the poster** as
well as to the list.
(Not to me - Oshel)
==========================================================
} Below is the result of your form, submitted on Wednesday, July 21,
} 2010 at 12:29:47 PM.
}
} realname - John Katahara
} Email - john_katahara-at-brown.edu
} EDUCATION - Graduate College
} SUBJECT_OF_QUESTION - Treating Polymer for TEM analysis
} QUESTION - I"m trying to get a TEM image of quantum dots embedded in
} polyacrylate polymer. I wanted to use a microtome to get thin
} sections, but the manager I contacted thought the resin might
} dissolve or in some way alter the acrylate.
}
} What would be the best way to go about treating the sample so that I
} can analyze it?
}

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From: ejb1176-at-gmail.com
Date: Wed, 21 Jul 2010 15:55:16 -0500
Subject: [Microscopy] FEI DB235 ion pump issues - seeking advice

Contents Retrieved from Microscopy Listserver Archives
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We are experiencing problems with starting the LMIG on our FIB after
experiencing a power outage last week.

We have been able to bring the E column and E gun back up and operational.
However, there is now a distinct clicking sound emanating from the I
column IGP (arcing?) when the IGP is energized.
Although the vacuum is within specs (3.5 to 3.8e10-9 range) it is
fluctuating slightly.

Attempts at running up the LMIG gun generate error messages and induce
larger column pressure changes (e-7 to e-9).
While running up the LMIG, almost continuous clicking sounds are heard.
Transient LMIG emission occurs but then the ion HT IGSU SPY intervenes
and shuts it down.

Short of venting the ion column and disassembling the IGP for
inspection and cleaning, or replacing the IGP completely, we are at a
loss for getting past this problem.
We are off contract due to budgetary reasons and pay as needed for FIB service.

If anyone has any tips for this type of problem, suggestions are appreciated.

Off-list responses preferred. I will post a summary of suggestions.....

thanks
ed

--
Edward Basgall, Ph.D.
Drexel University
Central Research Facility
office: (215) 895-2379
cell: (610) 574-4007

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From: sergey-at-seas.ucla.edu
Date: Wed, 21 Jul 2010 19:02:39 -0500
Subject: [Microscopy] viaWWW: old SEMs available

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Email: sergey-at-seas.ucla.edu
Name: Sergey Prikhodko

Organization: UCLA

Title-Subject: [Filtered] old SEMs

Message: Dear all,

We have two old SEMs we would like to get rid off 1) Cambridge
Stereoscan 250 and 2) Philips 525. Both are free (OBO) and should be
removed asap.

You can contact me directly.

Thanks,

Login Host: 128.97.83.27
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From: dsherman-at-purdue.edu
Date: Thu, 22 Jul 2010 13:45:22 -0500
Subject: [Microscopy] Low background holder for Philips CM microscope

Contents Retrieved from Microscopy Listserver Archives
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Hi all,

We are hunting for a low background holder for a CM-series microscope to use
for EDX. I would appreciate your contacting me if you have one you are not
using or plan to decommission a Philips CM microscope in the near future
that may free up one.

Thanks,
Debby


---
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy




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From: doc.vrdoljak-at-gmail.com
Date: Thu, 22 Jul 2010 14:38:42 -0500
Subject: [Microscopy] adapters to put standard pin mounts into Hitachi S4300 SEM

Contents Retrieved from Microscopy Listserver Archives
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Hello,
I'm having trouble figuring out which adapter that I need. I want to
be able to use a standard pin mount SEM stub inside of a Hitachi S4300
SEM which uses a screw on stub that mounts on a pin in the holder.
Looking over TedPella's website, there isn't a clear choice for me.
Anyone have a recommendation?

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1, 30 -- Subject: adapters to put standard pin mounts into Hitachi S4300 SEM
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From: wtivol-at-verizon.net
Date: Thu, 22 Jul 2010 14:57:12 -0500
Subject: [Microscopy] Re: Treating Polymer for TEM analysis

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}
} realname - John Katahara
} Email - john_katahara-at-brown.edu
} EDUCATION - Graduate College
} SUBJECT_OF_QUESTION - Treating Polymer for TEM analysis
} QUESTION - I"m trying to get a TEM image of quantum dots embedded in
} polyacrylate polymer. I wanted to use a microtome to get thin
} sections, but the manager I contacted thought the resin might
} dissolve or in some way alter the acrylate.
}
} What would be the best way to go about treating the sample so that I
} can analyze it?
}
Dear John,
You do not specify the form of the polyacrylate, but if it is a
block, I assume you would just have tried to section it. If the
polyacrylate is in small beads, which would need to be further
embedded in order to be sectioned, and if the shape of the
polyacrylate needs to be preserved (hence the manager's concern), one
possible solution is to embed the specimen in amorphous ice using a
high-pressure freezer and cryo-sectioning it. I think that there is
such equipment within a few hours drive of Brown, and it may be
possible to arrange to have your specimen prepared.
Yours,
Bill


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From: spurgeon-at-drexel.edu
Date: Thu, 22 Jul 2010 16:50:20 -0500
Subject: [Microscopy] FIB / LEAP - Preparation of MgO Nanopillars

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Hello everyone,

I am interested in preparing a thin film of Fe epitaxially grown on an
(001)-oriented MgO single crystal for local electrode atom probe
(LEAP). I have read studies in which authors etch an array of
nanopillars into the substrate, sputter their metal film onto the
pillars, and then directly proceed to LEAP. However, most of the
studies have been on Si and I have found little literature on creating
similar structures in MgO.

Is it possible to form ordered arrays of MgO nanopillars using a mask
in conjunction with etching/lithography? I'd like to create pillars
with a rather large aspect ratio (50-150 nm diameter vs. microns in
height). I have considered using a focused ion beam to mill them but
I'm worried about gallium implanation, as well as the length of time
needed to make them. Once I have the array it should (in principle) be
easy to deposit Fe and conduct LEAP measurements. I am worried about
comparing these results to a metal film deposited on a large, flat
substrate. Does anyone have experience with this?

Thank you for your help.
--
Steven Spurgeon
Dynamic Characterization Group
Department of Materials Science & Engineering
Drexel University

Cell: 719.330.0441
Office: Bossone 102
Email: spurgeon-at-drexel.edu
Web: http://www.materials.drexel.edu/dcg/

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From: tom_pella-at-tedpella.com
Date: Fri, 23 Jul 2010 12:53:26 -0500
Subject: [Microscopy] viaWWW: Bang Nguyen

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Email: sgkcck-at-aol.com
Name: Stacie Kirsch

Organization: Electron Microscopy Sciences

Title-Subject: [Filtered] Bang Nguyen

Message: It is with great sadness that we report the untimely passing
of our dear friend and colleague, Bang Nguyen. Bang was involved in
a fatal automobile accident on June 26 2010. We at Electron
Microscopy Sciences and Diatome have been devastated by the loss of
our friend of over 30 years.
On behalf of the entire staff of Electron Microscopy Sciences, thank
you for your support in this most difficult time.

Stacie Kirsch
Electron Microscopy Sciences


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8, 11 -- To: microscopy-at-microscopy.com
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From news-at-irinjalakudapressclub.com Fri Jul 23 05:52:45 2010
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Dear listers,

I have a potential customer with a Zeiss EM902A with EELS and STEM but no
schematics. Does anyone have a set that I might copy? I have the user's
manuals which will be digitized. Thanks.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz





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From kefxdarwindiz-at-xdarwin.com Fri Jul 23 11:01:15 2010
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Stacy,

I am so sorry to hear about the passing of Bang Nguyen. This is also a
loss to the microscopy community. I had become acquainted with Bang
through many exhibits we both attended. Bang was a person of integrity:
He was a gentleman, sharp as a tack, friendly even to his competitors, a
quick wit, skilled in his profession, stalwart in serving the needs both
of your company and the EM community. I respected him immensely.

I pray that you and your company, as well as his family, be comforted in
the mourning of this tragic loss.

Sincerely,

Tom Pella
General Manager
Ted Pella, Inc.
P.O. Box 492477
Redding, CA 96049
Tel: 800-237-3526
Fax: 530-243-3761
http://www.tedpella.com




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Email: sgkcck-at-aol.com
Name: Stacie Kirsch

Organization: Electron Microscopy Sciences

Title-Subject: [Filtered] Bang Nguyen

Message: It is with great sadness that we report the untimely passing of
our dear friend and colleague, Bang Nguyen. Bang was involved in a
fatal automobile accident on June 26 2010. We at Electron Microscopy
Sciences and Diatome have been devastated by the loss of our friend of
over 30 years.
On behalf of the entire staff of Electron Microscopy Sciences, thank you
for your support in this most difficult time.

Stacie Kirsch
Electron Microscopy Sciences


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From: rfklie-at-uic.edu
Date: Mon, 26 Jul 2010 10:42:16 -0500
Subject: [Microscopy] Post-doctoral position available

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

Dear Stacy,

Please extend our most sincere condolences from the entire MME staff to both our colleagues at EMS Diatome and to Bang's family. He will be warmly remembered as a treasured colleague.

Best regards,
Barbara Foster, President and Sr. Consultant

Microscopy/Microscopy Education
7101 Royal Glen Trail, Suite A
McKinney TX 75070
P: (972)924-5310 Skype: fostermme
W: www.MicroscopyEducation.com

At 08:20 PM 7/22/2010, sgkcck-at-aol.com wrote:



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From kefxastelloydiz-at-xastelloy.com Sat Jul 24 01:24:56 2010
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Dear Colleagues,

The Voyles Lab in the Department of Materials Science and Engineering at
the University of Wisconsin, Madison, seeks a post-doctoral research
associate to work on electron nanodiffraction of metal liquids. This
project will make extensive use of an aberration-corrected FEI Titan
STEM and other instruments in the UW Materials Science Center.
Extensive experience with materials characterization by TEM and STEM is
required; preference will be given to candidates with a background in
quantitative electron diffraction or in situ microscopy. A Ph.D. in
Materials Science, Physics, or related discipline is required.

For additional information, see tem.msae.wisc.edu and msc.engr.wisc.edu.
Duration is 1-3 years, and salary will be commensurate with
qualifications. To apply, please send a cover letter and CV including
contact information for three references to:

Paul Voyles
Materials Science and Engineering
1509 University Ave
Madison, WI 53706-1595
voyles-at-engr.wisc.edu
fax: 608-262-8353


==============================Original Headers==============================
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From kefxerosoxdiz-at-xerosox.com Sun Jul 25 16:34:40 2010
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Dear Colleagues,

A post-doctoral research associate position is available in the
Nanoscale Physics Group at the University of Illinois at Chicago to work
on the application of atomic-resolution Z-contrast imaging and electron
energy-loss spectroscopy (EELS) to thermoelectric oxide materials. This
position will be part of a larger collaboration between the University
of Illinois at Chicago (UIC) and the University of Alabama focusing on
the development of highly efficient high-temperature thermoelectric oxides.



The successful candidate will use atomic-resolution Z-contrast imaging
and electron energy-loss spectroscopy to determine the composition and
electronic structure of misfit-layered cobalt-oxides. Candidates should
have a Ph.D. in Physics, Materials Science or related disciplines. The
position requires extensive experience in transmission electron
microscopy and electron energy-loss spectroscopy. Experience working
with aberration-corrected scanning transmission electron microscopy is
also preferred. This position provides an opportunity to work on
cutting-edge research related to alternative energy production. UIC is
in the process of purchasing a new state-of-the-art aberration-corrected
cold field-emission scanning transmission electron microscope, and the
successful candidate will be directly involved in working with this new
instrument.

This postdoctoral position is available immediately, will be renewable
on an annual basis, and is anticipated for at least two years.
Preference will be given to recent PhD graduates who are currently in
the US and have a record of quality publication. Compensation will be
commensurate with qualification and experience. Health benefits will
also be provided. The position is open until it is filled.

Interested candidates can apply by email or by sending a cover letter,
CV with a complete list of publications, and contact information of 3
professional references to:

Professor Robert F Klie

Department of Physics

University ofIllinois atChicago

845 W Taylor Street, M/C 273

Chicago,IL 60607

email:rfklie-at-uic.edu



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From: Frank_Karl-at-lincolnelectric.com
Date: Mon, 26 Jul 2010 13:31:21 -0500
Subject: [Microscopy] measuring spot size

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


Hello everyone,
I want to run an little test on my JEOL 5800 SEM. I would like to measure
my spot size. Oh, sure the little spot size gauge works just fine , if you
like knowing that } } } } } } is smaller than } } } } } } } } } } .

I vaguely remember a test with a line scan and a Faraday trap but
everything else is gone. Any ideas on how I can actually measure the spot
size?

thanks in advance..........

Frank Karl
Lincoln Electric Co........

--
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From: jgbegl2-at-email.uky.edu
Date: Mon, 26 Jul 2010 14:27:32 -0500
Subject: [Microscopy] TEM processing of orchid seeds

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

Does anyone have experience with or a protocol for TEM processing of orchid seeds?  These are "immature" seeds so they are supposed to have a pore at one end that will allow liquid to enter, however, I still assume that prolonged fixation, washes, osmication, etc. would be the way to go.  If anyone has actual experience with them and could pass along some information that would be great.

Thanks,

Jim

Jim Begley
Research Facility Manager
Room 074 BBSRB
741 South Limestone Street
University of Kentucky
Lexington, KY 40506-0509
Phone: 859-323-2701
Fax: 859-257-1581
E-mail: jgbegl2-at-uky.edu

Other Contact:
Room 001 HSRB
Phone: 859-323-6108
Fax: 859-323-8089



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From: jdabbott-at-byu.net
Date: Mon, 26 Jul 2010 14:28:00 -0500
Subject: [Microscopy] Re: measuring spot size

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

As long as the diameter of the faraday cup aperture is bigger than
your beam spot size you measure the current in the faraday cup as a
function of beam position. Far from the cup edge the current should be
constant, at the edge of the cup the current will be changing as more
(or less) of the spot is in the cup. The width of that transition
region gives the width of the beam spot.


Jonathan Abbott
jdabbott-at-byu.net

It's kind of fun to do the impossible.
- Walt Disney

On Jul 26, 2010, at 12:43 PM, Frank_Karl-at-lincolnelectric.com wrote:

}
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}
} Hello everyone,
} I want to run an little test on my JEOL 5800 SEM. I would like to
} measure
} my spot size. Oh, sure the little spot size gauge works just fine ,
} if you
} like knowing that } } } } } } is smaller than } } } } } } } } } } .
}
} I vaguely remember a test with a line scan and a Faraday trap but
} everything else is gone. Any ideas on how I can actually measure
} the spot
} size?
}
} thanks in advance..........
}
} Frank Karl
} Lincoln Electric Co........
}
} --
} *************************************************************
} Note:
} The information contained in this message may be
} privileged and confidential and protected from disclosure. If
} the reader of this message is not the intended recipient, or
} an employee or agent responsible for delivering this message
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} dissemination, distribution or copying of this communication
} is strictly prohibited. If you have received this
} communication in error, please notify us immediately by
} replying to the message and deleting it from your computer.
} Thank you,
} The Lincoln Electric Company
} **************************************************************
}
}
} ==============================Original
} Headers==============================
} 7, 21 -- From frank_karl-at-lincolnelectric.com Mon Jul 26 13:31:20 2010
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} 7, 21 -- From: Frank_Karl-at-lincolnelectric.com
} 7, 21 -- X-MIMETrack: CD-MIME by Router on Notescom1/Lincoln
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From: amsamuel-at-uqac.ca
Date: Mon, 26 Jul 2010 19:08:05 -0500
Subject: [Microscopy] viaWWW: Accessory needed for Phillips XL-120 SEM

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Email: amsamuel-at-uqac.ca
Name: A.M. Samuel

Organization: University of Quebec at Chicoutimi (UQAC)

Title-Subject: [Filtered] Accessory needed for Phillips XL-120 SEM

Message: Hello,

I would like to find out if you know of anyone
who can supply a computer video connector board
for use with a Phillips XL-120 scanning electron
microscope? My PhD supervisor in India asked me
if I could manage to get it, as it is made in
Canada, and their suppliers in India ñ the
Phillips EM Co. Eindhoven is now dissolved, so
they have no means of getting it.

The part needed is the Oculus-TCX/MX Part number OC-TCXM-INST0 Edition 2.02.

I would be grateful to received any possible leads.

Thanks in advance.

A.M. Samuel



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From: ech-at-uvic.ca
Date: Mon, 26 Jul 2010 19:08:39 -0500
Subject: [Microscopy] viaWWW: the plot thickens: Family Affair

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Email: ech-at-uvic.ca
Name: Elaine Humphrey

Organization: University of Victoria

Title-Subject: [Filtered] the plot thickens: Family Affair

Message: The story so far!

CSI Portland: The Family Affair at M&M

A man walked in to the Oregon Museum of Science
and Industry (OSMI) gift store and passed an
envelope over to the volunteer behind the
counter. On the envelope it said ìPut all the
money in the till in the envelope. Don't say
anything, I've got a gun!î She complied as she
has been told to do if something like this
happens but as soon as he had gone, she called
the police. She told the policeman that she saw
him go off on a bicycle. As he rode away he
scraped against a rock with his bike and left
some metal scrapings behind. He also left some
thread from his trousers and a bolt broke off
from the bike. In hurrying to escape, he rode
through a patch of sandy building material and
over a bed of flowers.

That evening the police had six persons in
custody all with bikes with scraped sides and
sandy wheels and their trousers were dusted with
pollen. They each had dubious alibis easily
proved or disproved with microscopy.

Family Affair is for families and friends of
delegates to have a fun time with microscopy and
a chance to visit the Exhibition Hall. As well as
participating in "CSI Portland," other activities
include "Microscopic Explorations" which uses
microscopy to take a closer look at everyday
objects and whatever live critters volunteered
that morning! Wednesday 1.30 pm room B112


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From: gary-at-gaugler.com
Date: Mon, 26 Jul 2010 19:26:17 -0500
Subject: [Microscopy] Re: viaWWW: Accessory needed for Phillips XL-120

Contents Retrieved from Microscopy Listserver Archives
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Try here:

http://www.murphyjunk.bizland.com/id29.html

$389US. Not sure if it is the revision you need.

gary g.


At 05:10 PM 7/26/2010, you wrote:



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From: r.sims-at-auckland.ac.nz
Date: Mon, 26 Jul 2010 19:52:35 -0500
Subject: [Microscopy] measuring spot size

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Unfortunately, that's not the only necessary condition.

The edge of the aperture (or of whatever other contrast edge you're planning on using) has to be known independently to be sharp, without any rounding to at least a tenth or so of the spot diameter that you're trying to measure.

Any suggestions as to what to use will be gratefully received.
cheers
Ritchie


On 27 Jul 2010 at 7:28, jdabbott-at-byu.net wrote:


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Jonathan Abbott
jdabbott-at-byu.net
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- Walt Disney
On Jul 26, 2010, at 12:43 PM, Frank_Karl-at-lincolnelectric.com wrote:
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} Hello everyone,
} I want to run an little test on my JEOL 5800 SEM. I would like to
} measure
} my spot size. Oh, sure the little spot size gauge works just fine ,
} if you
} like knowing that } } } } } } is smaller than } } } } } } } } } } .
}
} I vaguely remember a test with a line scan and a Faraday trap but
} everything else is gone. Any ideas on how I can actually measure
} the spot
} size?
}
} thanks in advance..........
}
} Frank Karl
} Lincoln Electric Co........
}

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10, 47 -- Subject: Re: [Microscopy] Re: measuring spot size
10, 47 -- Thread-Topic: [Microscopy] Re: measuring spot size
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From: zaluzec-at-aaem.amc.anl.gov
Date: Mon, 26 Jul 2010 20:15:02 -0500
Subject: [Microscopy] Re: MgO cubes for measuring spot size

Contents Retrieved from Microscopy Listserver Archives
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On the STEM side of things , I have used Mg burned to form MgO smoke and captured with a holey carbon film to measure larger (nm size) probes.

The MgO "smoke" particles are generally in the form of very small cubes. You can easily find a few that are nearly atomically sharp, at least to the extent that you can use them to measure the signal rise profile.

Of course, I've only used this for larger STEM probes not SEM, but if you have a STEM attachment in your SEM it should also work. In purley SEM mode the contrast profile will not be so advantageous as you can get secondary emission from both the top and bottom surfaces of the cube depending upon, your accelerating voltage, the cube size and how you have tilted it to orient its edges.

Remember to wear the appropriate safety goggles and associated equipment when burning the Mg, and only use very very small amounts of Mg. i.e. check your MSDS documentation. Lastly the MgO doesn't last long in humid environments.

Nestor
Your Friendly Neighborhood SysOp







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}
}
} Unfortunately, that's not the only necessary condition.
}
} The edge of the aperture (or of whatever other contrast edge you're planning on using) has to be known independently to be sharp, without any rounding to at least a tenth or so of the spot diameter that you're trying to measure.
}
} Any suggestions as to what to use will be gratefully received.
} cheers
} Ritchie
}
}
} On 27 Jul 2010 at 7:28, jdabbott-at-byu.net wrote:
}
}
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} As long as the diameter of the faraday cup aperture is bigger than
} your beam spot size you measure the current in the faraday cup as a
} function of beam position. Far from the cup edge the current should be
} constant, at the edge of the cup the current will be changing as more
} (or less) of the spot is in the cup. The width of that transition
} region gives the width of the beam spot.
}
} Jonathan Abbott
} jdabbott-at-byu.net
} It's kind of fun to do the impossible.
} - Walt Disney
} On Jul 26, 2010, at 12:43 PM, Frank_Karl-at-lincolnelectric.com wrote:
} }
} }
} }
} }
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} --------------------------------------------------------------------------
} --
} }
} }
} } Hello everyone,
} } I want to run an little test on my JEOL 5800 SEM. I would like to
} } measure
} } my spot size. Oh, sure the little spot size gauge works just fine ,
} } if you
} } like knowing that } } } } } } is smaller than } } } } } } } } } } .
} }
} } I vaguely remember a test with a line scan and a Faraday trap but
} } everything else is gone. Any ideas on how I can actually measure
} } the spot
} } size?
} }
} } thanks in advance..........
} }
} } Frank Karl
} } Lincoln Electric Co........
} }
}
} ==============================Original Headers==============================
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} 10, 47 -- Date: Tue, 27 Jul 2010 12:52:16 +1200
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===========================================
Dr. Nestor J. Zaluzec
Argonne National Laboratory
Electron Microscopy Center
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Senior Scientist - Argonne National Laboratory
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Senior Fellow of the Computational Institute - University of Chicago
E.P. Wigner Fellow - Oak Ridge National Laboratory
President-Elect: Microscopy Society of America
===========================================






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From: vray-at-partbeamsystech.com
Date: Mon, 26 Jul 2010 23:25:29 -0500
Subject: [Microscopy] Re: measuring spot size

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

Came up in Google:

http://www.abeamtech.com/?dir=products/BEAMETR&pg=about

No any affiliation here.

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com

On 7/26/2010 8:53 PM, r.sims-at-auckland.ac.nz wrote:
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} Unfortunately, that's not the only necessary condition.
}
} The edge of the aperture (or of whatever other contrast edge you're planning on using) has to be known independently to be sharp, without any rounding to at least a tenth or so of the spot diameter that you're trying to measure.
}
} Any suggestions as to what to use will be gratefully received.
} cheers
} Ritchie
}
}
} On 27 Jul 2010 at 7:28, jdabbott-at-byu.net wrote:
}
}
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} As long as the diameter of the faraday cup aperture is bigger than
} your beam spot size you measure the current in the faraday cup as a
} function of beam position. Far from the cup edge the current should be
} constant, at the edge of the cup the current will be changing as more
} (or less) of the spot is in the cup. The width of that transition
} region gives the width of the beam spot.
}
} Jonathan Abbott
} jdabbott-at-byu.net
} It's kind of fun to do the impossible.
} - Walt Disney
} On Jul 26, 2010, at 12:43 PM, Frank_Karl-at-lincolnelectric.com wrote:
} }
} }
} }
} }
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} }
} }
} } Hello everyone,
} } I want to run an little test on my JEOL 5800 SEM. I would like to
} } measure
} } my spot size. Oh, sure the little spot size gauge works just fine ,
} } if you
} } like knowing that} } } } } } is smaller than} } } } } } } } } } .
} }
} } I vaguely remember a test with a line scan and a Faraday trap but
} } everything else is gone. Any ideas on how I can actually measure
} } the spot
} } size?
} }
} } thanks in advance..........
} }
} } Frank Karl
} } Lincoln Electric Co........
} }
}
} ==============================Original Headers==============================
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From: oshel1pe-at-cmich.edu
Date: Tue, 27 Jul 2010 08:04:34 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist

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



ASTM E986 - Standard
Practice for Scanning Electron Microscope Beam Size Characterization gives
a procedure using the Y-deflection waveform which would be the line-scan
image mode on the JSM 5800.

Larry D. Hanke,
P.E.
Materials Evaluation and Engineering, Inc.
Practical
Solutions Through Technology and Innovation
http://www.mee-inc.com
(763) 449-8870


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} Hello everyone,
} I want to run an
little test on my JEOL 5800 SEM. I would like to measure
} my
spot size. Oh, sure the little spot size gauge works just fine , if
} you
} like knowing that } } } } } } is smaller
than } } } } } } } } } } .
}
} I
vaguely remember a test with a line scan and a Faraday trap but
}
everything else is gone. Any ideas on how I can actually measure the
spot
} size?
}
} thanks in advance..........
}
} Frank Karl
} Lincoln Electric Co........
}
} --
}
*************************************************************
}
Note:
} The information contained in this message may be
} privileged and confidential and protected from disclosure. If
} the reader of this message is not the intended recipient, or
} an employee or agent responsible for delivering this message
} to the intended recipient, you are hereby notified that any
} dissemination, distribution or copying of this communication
} is strictly prohibited. If you have received this
}
communication in error, please notify us immediately by
}
replying to the message and deleting it from your computer.
}
Thank you,
} The Lincoln Electric Company
}
**************************************************************
}

}
} ==============================Original
}
Headers==============================
} 7, 21 --
X-from
frank_karl-at-lincolnelectric.com Mon Jul 26 13:31:20 2010
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} Date: Mon, 26 Jul 2010 16:03:04 -0700 (PDT)
} From: Andrew Kustas {abkustas-at-rams.colostate.edu}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Monday, July 26, 2010
} at 04:02:57 PM.
}
} realname - Andrew Kustas
} Email - abkustas-at-rams.colostate.edu
} ORGANIZATION - Colorado State University
} EDUCATION - Undergraduate College
} LOCATION - Parker, CO, 80521
} SUBJECT_OF_QUESTION - Cleaving sample
} QUESTION - Hello, I am trying to view the thickness of a Si/SiC
} superlattice that I have been laying down on some Silicon wafers.
} The images from the SEM show a messy surface (debris of material)
} near the superlattice as a result of the method used to break the
} silicon along its cross-section. The current method I use is a
} diamond scribe to scratch the backside of the substrate and then a
} corresponding force to break the silicon piece along the scribed
} line. What is a good method for breaking this silicon substrate to
} view the cross-section and get adequately clear images of the
} coating thickness without having much debris from the break?
}
--
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From: jwheckman-at-earthlink.net
Date: Tue, 27 Jul 2010 08:52:23 -0500
Subject: [Microscopy] RE: Cleaving sample

Contents Retrieved from Microscopy Listserver Archives
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Andrew-
A method that has worked on occasion for me is to scribe (diamond scribe) a
short line ( 4-5 mm) on the front side oriented, as best as I can eye-ball,
to a {100} . Then cleave with firm but constantly increasing pressure
(surprise break style) on a pair of glazier's pliers (e.g. Fletcher-Terry
Co 6" Glass Cutter Pliers No. 06-111). For smaller pieces, just a nick on
the edge with thumb pressure at the ends and a piece of W wire as a fulcrum
works well. This approach works well with a number of thin (and not so
thin) films on Si wafers.
Good luck.
John


} [Original Message]
} From: {oshel1pe-at-cmich.edu}
} To: {jwheckman-at-earthlink.net}
} Date: 7/27/2010 6:09:41 AM
} Subject: [Microscopy] Fwd: Ask-A-Microscopist
}
}
}
}
}
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} } Date: Mon, 26 Jul 2010 16:03:04 -0700 (PDT)
} } From: Andrew Kustas {abkustas-at-rams.colostate.edu}
} } To: oshel1pe-at-cmich.edu
} } Subject: Ask-A-Microscopist
} }
} } Below is the result of your form, submitted on Monday, July 26, 2010
} } at 04:02:57 PM.
} }
} } realname - Andrew Kustas
} } Email - abkustas-at-rams.colostate.edu
} } ORGANIZATION - Colorado State University
} } EDUCATION - Undergraduate College
} } LOCATION - Parker, CO, 80521
} } SUBJECT_OF_QUESTION - Cleaving sample
} } QUESTION - Hello, I am trying to view the thickness of a Si/SiC
} } superlattice that I have been laying down on some Silicon wafers.
} } The images from the SEM show a messy surface (debris of material)
} } near the superlattice as a result of the method used to break the
} } silicon along its cross-section. The current method I use is a
} } diamond scribe to scratch the backside of the substrate and then a
} } corresponding force to break the silicon piece along the scribed
} } line. What is a good method for breaking this silicon substrate to
} } view the cross-section and get adequately clear images of the
} } coating thickness without having much debris from the break?
} }
} --
}
****************************************************************************
*****************
} Forwarded from "Ask a Microscopist"
} Please remember that the person asking the question is likely not a
} member the listserver, and **any reply should go directly to the
} poster** as well as to the list.
} Using the "reply" function in your email does *not* send your answer
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}
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*****************
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} ==============================Original
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From: jmircheski-at-us.es
Date: Thu, 29 Jul 2010 09:18:12 -0500
Subject: [Microscopy] TEM reusing OsO4

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

Dear microscopists,
Our CM200 STEM still runs quite well, but our Emispec digital beam control + spectrum imaging system is rapidly reaching the end of its life. Does anyone have any experience or insight regarding putting modern analytical control onto a CM-series STEM?
The two possibilities I've investigated are EDAX Genesis (since we have a relatively new EDAX Si(Li) in place) and Gatan Digiscan (for EDS and EELS spectrum imaging, plus filter control).
I'd appreciate any insight, advice, or comments the community may have on this topic.

Thanks
Chad Parish ( parishcm-at-ornl.gov )


==============================Original Headers==============================
3, 34 -- From parishcm-at-ornl.gov Tue Jul 27 14:25:02 2010
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3, 34 -- From: "Parish, Chad M." {parishcm-at-ornl.gov}
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From jadher.dominguez-at-unops.org Tue Jul 27 14:53:41 2010
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Dear listers,
Below is the repeat announcement for a postdoctoral position to start
October 1 (can be earlier if needed). The work involves in-situ
microscopy as well as high resolution aberration corrected STEM and
EELS. Please forward to any potentially interested parties.
Albina Borisevich


Combined Scanning Transmission and Scanning Probe Microscopy of Oxides

Materials Science and Technology Division

Oak Ridge National Laboratory

Project Description:

The Materials Science and Technology Division at Oak Ridge National
Laboratory (ORNL) is seeking a candidate to fill a postdoctoral position
in the field of combined scanning transmission electron microscopy and
scanning probe microscopy of oxide thin films and nanostructures. The
position is available October 1 2010. This program takes advantage of
ORNL’s suite of advanced electron microscopes, including 5
aberration-corrected instruments, as well as (S)TEM/STM and (S)TEM/AFM
capabilities.

The successful applicant must demonstrate experience in electron
microscope operation, preferably FEI microscopes, as well as skills in
analysis and interpretation of microscopic and spectroscopic data. This
position provides an opportunity to join an experienced team working in
a highly collaborative environment. Interactions with the scanning probe
microscopy program at ORNL’s Center for Nanophase Materials Sciences
(CNMS) are anticipated on a daily basis.

Qualifications: PhD degree required

This position requires a Ph.D. in Materials Science, Physics, or related
field, with an emphasis on advanced TEM or STEM. Knowledge of oxide
crystal chemistry is a plus. Excellent oral and written communication
skills are required, and presentations and publication of scientific
results in peer-reviewed journals are expected. The applicant must have
the ability to work in a team and interact effectively with a broad
range of colleagues. Applicants cannot have received the most recent
degree more than five years prior to the date of application and must
complete all degree requirements before starting their appointment.

Technical Questions:

Questions regarding the position can be directed to Drs. Albina Y.
Borisevich, albinab-at-ornl.gov, Stephen J. Pennycook,
pennycooksj-at-ornl.gov, and Sergei V. Kalinin, sergei2-at-ornl.gov. Please
include the requisition number and title when corresponding.

How to Apply:

Qualified applicants must apply online at
https://www2.orau.gov/ORNL_POST/. All applicants will need to register
before they can begin the online application. For complete instructions,
on how to apply, please see the instructions at
http://www.orau.gov/orise/edu/ornl/ornl-pdpm/application.htm.

This appointment is offered through the ORNL Postgraduate Research
Participation Program and is administered by the Oak Ridge Institute for
Science and Education (ORISE). The program is open to all qualified U.S.
and non-U.S. citizens without regard to race, color, age, religion, sex,
national origin, physical or mental disability, or status as a
Vietnam-era veteran or disabled veteran.
--
Albina Y. Borisevich
R&D Staff
Electron Microscopy Group
Oak Ridge National Laboratory
Materials Science and Technology Division
PO Box 2008
Oak Ridge TN 37831-6031

http://stem.ornl.gov/

For express mail add: 1 Bethel Valley Road
phone: (865) 576-4060
fax: (865) 574-4143

==============================Original Headers==============================
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From sales-us-at-iopinc.com Wed Jul 28 03:37:30 2010
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} Below is the result of your form, submitted on Wednesday, July 28,
} 2010 at 12:12:29 PM.
}
} realname - Mark Twigg
} Email - twigg-at-estd.nrl.navy.mil
} EDUCATION - Graduate College
} LOCATION - Washington, DC
} SUBJECT_OF_QUESTION - HRTEM Siting Near Laser
} QUESTION - My organization is slated to be moved to a new building
} sometime in the next several years. Of course that raises the
} question concerning where to site my HRTEM. Currently I have a top
} entry Hitachi H9000UHR. By the time this building is ready (maybe
} six years in the future) we may have a new HRTEM instrument with
} more demanding requirements. The problem is that the floor plan for
} the building has just been altered with my lab positioned right next
} to an optics lab with a high-powered laser designed to blast
} electronic devices as a way of simulating radiation effects.
} Anybody with horror tales about living next door to one of these
} things?
}
} Thanks,
}
} Mark
--
***************************************************************************************
Forwarded from "Ask a Microscopist"
Please remember that the person asking the question is likely
not a member the listserver, and
**any reply should go directly to the poster**
as well as to the list.
Using the "reply" function in your email does *not* send your answer
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Please copy their email address from their question.
****************************************************************************************

==============================Original Headers==============================
1, 23 -- From oshel1pe-at-cmich.edu Wed Jul 28 14:29:21 2010
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From mist-at-csumb.edu Thu Jul 29 00:04:11 2010
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Message-ID: {01cb2f0d$71c4e300$6ff1bccb-at-mist}

Dear microscopists,

I am currently performing negative staining with lipid vesicles and
sometimes I get dark circular shadows, about 100-200nm in size which seem to
lie above my vesicles. We are wondering if this can be caused
by high air humidity during the drying process, since the weather was quite
hot and moist during the last weeks. Has anyone an idea about the reason for this phenomenon and how to get rid of that?

Thanks in advance,
Kerstin












--
GMX.at - Österreichs FreeMail-Dienst mit über 2 Mio Mitgliedern
E-Mail, SMS & mehr! Kostenlos: http://portal.gmx.net/de/go/atfreemail

==============================Original Headers==============================
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From intern-at-marineland.net Thu Jul 29 06:47:11 2010
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Hi All,

I have some memory of reading somewhere or hearing from someone that the
OsO4 (used for fixing TEM samples) can be reused by adding some hydrogen
peroxide. Could someone tell me if this is true and if anyone has experience
in reusing the OsO4? It is quite valuable, so if this is true I’d like to
save as much as possible of it.
Looking forward to your information!

Regards,

Dr. Josif Mircheski
____________________________________________________________________________
___
Instituto de Biomedicina de Sevilla (IBIS)
Hosp.Univ. Virgen del Rocío/CSIC/Univ. de Sevilla y
Dpto. Fisiologia Médica y Biofísica
Universidad de Sevilla
Facultad de Medicina
Avda. Sánchez-Pizjuán 4
41009-Sevilla
 
Phone:+34-954556103
Fax:+34-954551769
e-mail: jmircheski-at-us.es





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From: larry.ackerman-at-ucsf.edu
Date: Thu, 29 Jul 2010 12:06:29 -0500
Subject: [Microscopy] Re: Sterility of organisms in Epoxy

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

Cells and microorganisms are the predominate samples in biological
microscopy but there are sub-cellular components that may present a
health and safety issue. In particular, prions are not easily
inactivated and when they are the tissue is useless for microscopy. If I
have such a sample I do not grind it or saw it which might create
potentially "infectious" dust. I also avoid direct contact with
sections. Maybe in 20 years we will know the efficacy of these procedures.

Larry

reinhard.rachel-at-biologie.uni-regensburg.de wrote:
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} Although this issue was raised early in June, I still think it is ok to post my opinion, my reply - which was left "unsent" in the folder "in progress" ...
}
} } Message: Has anyone had dealings with the USDA, CDC, or HHS, or other
} } agencies with respect to proving sterility of agents that have been
} } fixed (4%paraformaldehyde/1%gluteraldehyde), osmicated(1% osmium),
} } (potentially irradiated 2X16[6] rads), dehydrated, and embedded in
} } epoxy.
} }
} } I find myself getting questions about things that never were
} } questioned in the past.
}
} as a microbiologist and EM specialist in Germany, I am lucky that nobody raised this question to me, so far (no USDA, CDC, HHS, ...).
} I would bet that no (or hardly any?) microorganism = cell will survive the usual preparation / embedding protocol (usual: at RT). But who knows?!? what about viruses, spores? extremophiles? I am working with those extremophiles since 20 years now, none of which was so far shown to be infectious to human beings. In all studies, we were and are surprised to learn what they do tolerate.
} For me, the worst step in EM preparation is not necessarily, or not only, the FA/GA and Os fixation/oxidation (do these substances fully penetrate and react with those molecules which are "important" for infectivity? in viruses, in spores, in vegetative cells?), but in particular the stepwise dehydration at RT in Ethanol or Acetone, and the final infiltration of the (hydrophobic, monomeric) resin. During these steps, most (all?) cells are very likely (!?!) to be destroyed, as the hydrophobic part of the membranes, the lipids, are extracted to great extent, and the membrane is not intact any more. During the final polymerization, almost all (?) bio-molecules are altered, inevitably.
} But, there are question marks; and, what happens to the cells during freeze substitution? during embedding in LR white or gold, or Lowicryls? we electron microscopists are happy about the good, much improved structural preservation. Full stop. Does this - vice versa - mean that (few of) these cells are in a status to survive? at least their DNA? some mRNA? how to test this? And what about those few cells which were not fully embedded in the resin, due to "infiltration problems", and fall out of the section during the sectioning process? is this "cell" / cell debris / protein / ?? potentially infectious, or of harm to the operator or experimentator? How to test this, who wants to test this?
}
} I do not want to make the sitution complicated. Understanding of all these processes is necessary, awareness is needed. - For me, personally, we have to protect ourselves from all the dangerous chemicals (FA, GA, Os, resin, resin dust, etc.). There is the real hazard potential, everyday.
}
} And, yes, my bet is that not a single cell does survive, in particular the "routine embedding at room temperature". - But, which microorganism, which officer, takes care of my bet?
}
} kind regards,
} Reinhard
}
}

--
Larry Ackerman, Specialist
Electron Microscopy Lab
Manager, DERC Microscopy Core
UCSF, Dept. of Anatomy, Rm S1355
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

415-999-4758

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From: rcmoretz-at-gmail.com
Date: Thu, 29 Jul 2010 14:05:19 -0500
Subject: [Microscopy] Re: TEM reusing OsO4

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

There is a lot of old literature on this from the 1960s & 70s.
Unfortunately I no longer have the references as they evaporated along
with all of my reprints when I retired. A search of PubMed should
bring up some references.

Roger Moretz, Ph.D.

On Thu, Jul 29, 2010 at 10:23 AM, {jmircheski-at-us.es} wrote:
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} Hi All,
}
} I have some memory of reading somewhere or hearing from someone that the
} OsO4 (used for fixing TEM samples) can be reused by adding some hydrogen
} peroxide. Could someone tell me if this is true and if anyone has experience
} in reusing the OsO4? It is quite valuable, so if this is true I’d like to
} save as much as possible of it.
} Looking forward to your information!
}
} Regards,
}
} Dr. Josif Mircheski
} ____________________________________________________________________________
} ___
} Instituto de Biomedicina de Sevilla (IBIS)
} Hosp.Univ. Virgen del Rocío/CSIC/Univ. de Sevilla y
} Dpto. Fisiologia Médica y Biofísica
} Universidad de Sevilla
} Facultad de Medicina
} Avda. Sánchez-Pizjuán 4
} 41009-Sevilla
}
} Phone:+34-954556103
} Fax:+34-954551769
} e-mail: jmircheski-at-us.es
}
}
}
}
}
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} 8, 39 -- Subject: TEM reusing OsO4
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From: rcmoretz-at-gmail.com
Date: Thu, 29 Jul 2010 14:15:12 -0500
Subject: [Microscopy] Sterility of organisms in Epoxy

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

I always have understood that once biological materials are exposed to
OsO4 that they are totally inactivated. When it comes to "sawing"
epoxy blocks I am more concerned with the dust that is created, not
the organisms in the epoxy. I worked with prions for years & never
had the fear of prions I see expressed today. Simply handle with
care--but OsO4 will inactivate prions as surely as it inactivates any
other biological organism. My opinion.

Roger Moretz Ph.D.

On Thu, Jul 29, 2010 at 1:10 PM, {larry.ackerman-at-ucsf.edu} wrote:
}
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} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
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} ----------------------------------------------------------------------------
}
} Cells and microorganisms are the predominate samples in biological
} microscopy but there are sub-cellular components that may present a
} health and safety issue. In particular, prions are not easily
} inactivated and when they are the tissue is useless for microscopy. If I
} have such a sample I do not grind it or saw it which might create
} potentially "infectious" dust.  I also avoid direct contact with
} sections. Maybe in 20 years we will know the efficacy of these procedures.
}
} Larry
}
} reinhard.rachel-at-biologie.uni-regensburg.de wrote:
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} } To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } Although this issue was raised early in June, I still think it is ok to post my opinion, my reply - which was left "unsent" in the folder "in progress" ...
} }
} } } Message: Has anyone had dealings with the USDA, CDC, or HHS, or other
} } } agencies with respect to proving sterility of agents that have been
} } } fixed (4%paraformaldehyde/1%gluteraldehyde), osmicated(1% osmium),
} } } (potentially irradiated 2X16[6] rads), dehydrated, and embedded in
} } } epoxy.
} } }
} } } I find myself getting questions about things that never were
} } } questioned in the past.
} }
} } as a microbiologist and EM specialist in Germany, I am lucky that nobody raised this question to me, so far (no USDA, CDC, HHS, ...).
} } I would bet that no (or hardly any?) microorganism = cell will survive the usual preparation / embedding protocol (usual: at RT). But who knows?!? what about viruses, spores? extremophiles? I am working with those extremophiles since 20 years now, none of which was so far shown to be infectious to human beings. In all studies, we were and are surprised to learn what they do tolerate.
} } For me, the worst step in EM preparation is not necessarily, or not only, the FA/GA and Os fixation/oxidation (do these substances fully penetrate and react with those molecules which are "important" for infectivity? in viruses, in spores, in vegetative cells?), but in particular the stepwise dehydration at RT in Ethanol or Acetone, and the final infiltration of the (hydrophobic, monomeric) resin. During these steps, most (all?) cells are very likely (!?!) to be destroyed, as the hydrophobic part of the membranes, the lipids, are extracted to great extent, and the membrane is not intact any more. During the final polymerization, almost all (?) bio-molecules are altered, inevitably.
} } But, there are question marks; and, what happens to the cells during freeze substitution? during embedding in LR white or gold, or Lowicryls? we electron microscopists are happy about the good, much improved structural preservation. Full stop. Does this - vice versa - mean that (few of) these cells are in a status to survive? at least their DNA? some mRNA? how to test this? And what about those few cells which were not fully embedded in the resin, due to "infiltration problems", and fall out of the section during the sectioning process? is this "cell" / cell debris / protein / ?? potentially infectious, or of harm to the operator or experimentator? How to test this, who wants to test this?
} }
} } I do not want to make the sitution complicated. Understanding of all these processes is necessary, awareness is needed. - For me, personally, we have to protect ourselves from all the dangerous chemicals (FA, GA, Os, resin, resin dust, etc.). There is the real hazard potential, everyday.
} }
} } And, yes, my bet is that not a single cell does survive, in particular the "routine embedding at room temperature". - But, which microorganism, which officer, takes care of my bet?
} }
} } kind regards,
} } Reinhard
} }
} }
}
} --
} Larry Ackerman, Specialist
} Electron Microscopy Lab
} Manager, DERC Microscopy Core
} UCSF, Dept. of Anatomy, Rm S1355
} 513 Parnassus Ave., Box 0452
} San Francisco, CA 94143
}
} larry.ackerman-at-ucsf.edu
}
} 415-999-4758
}
} ==============================Original Headers==============================
} 6, 36 -- From btv1==826201a3f60==Larry.Ackerman-at-ucsf.edu Thu Jul 29 12:06:29 2010
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From: joe.kintz-at-stress.com
Date: Thu, 29 Jul 2010 14:57:24 -0500
Subject: [Microscopy] viaWWW: Job Opportunity for SEM Operator/Metallurgical Technician

Contents Retrieved from Microscopy Listserver Archives
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This Question/Comment was submitted to the Microscopy Listserver
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Email: joe.kintz-at-stress.com
Name: Joseph Kintz

Organization: Stress Engineering Services, Inc.

Title-Subject: [Filtered] Job Opportunity for SEM
Operator/Metallurgical Technician

Message: Stress Engineering Services, Inc. in
northwest Houston, TX is seeking an SEM operator
and metallurgical technician. The ideal
candidate will have at least 10 years experience
performing scanning electron microscopy of metals
and nonmetals for a commercial laboratory. He or
she will also have at least 10 years experience
performing metallography, photography, and other
metallurgical tests and evaluations for a
commercial laboratory.

If youíre interested and meet these
qualifications, please send me your resume and
three references. No phone calls please.

Joe


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From: Phil.Ahrenkiel-at-sdsmt.edu
Date: Thu, 29 Jul 2010 18:22:20 -0500
Subject: [Microscopy] floor stability for TEM

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

It is clear from the evolution of the methodology by which microorganisms are treated for viewing with both light and electron microscopes that it (the methodology) was and remains intended for membrane-bound organisms (included would be membrane encapsulated virions). The aldehydes and osmium surely have extremely deleterious effects on living cells and organisms - even prokaryotes and archaea. But the words 'killing', 'fixing' and 'sterilizing' each has its own definition. Achieving one does not necessarily achieve them all.

The question that bears on prions should be considered in the context of CDC warnings on the subject buried in the page at the URL below.
URL: http://www.cdc.gov/ncidod/eid/vol9no3/02-0327.htm

The dust from sawing plastics/resins has always been 'suggested' to be hazardous, so......

Inactivation vs. sterility is another matter altogether and should be addressed by an expert.

To be blunt, it is the responsibility of the laboratory director and higher administrators to generate policies for handling various chemical, material, and biological specimens - especially where they are all investigated in the same core facility. I have yet to meet the PhD (or other) who is sufficiently adept to discount the potential hazards of work in such environments as those found in chemical, material and biological laboratories, and for the most part, these are NOT deluged with specimens characterized as highly hazardous. A PhD is a certificate of intelligence, but the greatest lessons of that journey are insights gained from the knowledge of what is NOT known by the holder rather than the expertise in one small niche - no matter its immediate or future potential.

Sterility is as variable as the means by which its achievement can be claimed/alleged. Thus, it is always to be mistrusted.

Regards to all,

Fred Monson


Frederick C. Monson, PhD
Tecnhical Director
Center for Microanalysis and Imaging Research and Training Center (CMIRT)
Large Scientific Instrument Core
West Chester University S. Church St. and W. Rosedale Ave.
West Chester, PA, 19383
610-738-0437

Frederick C. Monson, PhD
Tecnhical Director
Center for Microanalysis and Imaging Research and Training Center (CMIRT)
Large Scientific Instrument Core
West Chester University S. Church St. and W. Rosedale Ave.
West Chester, PA, 19383
610-738-0437
fmonson-at-wcupa.edu



==============================Original Headers==============================
13, 25 -- From FMonson-at-wcupa.edu Thu Jul 29 15:37:05 2010
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13, 25 -- Subject: Re: sterility of organisms in Epoxy
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From mississauga-at-roberthalftechnology.com Thu Jul 29 15:38:06 2010
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We will be installing a new 200-KV LaB6 TEM within the next few months, and our university has allocated a nice lab in the basement of our older (ca. 1972) building for the instrument. But the environmental survey showed an on-going, 18-Hz, 25-um vertical oscillation in that room due to the building's HVAC. Also, the floor lacks resiliency in some areas, maybe due to shifting over time of the soil below, so occasional, nearby foot traffic could affect image stability.

The survey company offers an active vibration-cancellation system to remove the low-frequency noise. Though I am not opposed to this, my first priority is to address any issues with the building foundation before installation. So I have spoken with several contractors about the possibility of forming an isolated region of the floor to support the TEM.

My understanding is that the vibration propagates readily through the rigid cement floor, which lacks sufficient mass to damp the oscillation. It would seem that the ideal floor has 1) large mass and 2) weak coupling to the driving force.

One practical possibility I investigated is to install a detached, separate section of floor on concrete piers, set deep (12-20 ft) into the ground, upon which the TEM column would rest. Now, I am not a civil/mechanical engineer, but my naïve impression is that if the piers extend deep enough, the earth would essentially act as a massive load. The underlying soil is relatively soft, so the vibrations from the building would not substantially propagate through the earth from the building to the TEM. Plus, this should decouple the column from the higher-frequency traffic in the building.

The employee of the microscope manufacturer with whom I spoke was adamant that this was a ridiculous idea, and would actually make the vibration worse. He believes it would require an impractically massive foundation to impede these oscillations, that the earth would not damp the vibration, and that the active cancellation system is our only option.

Of course, we will consult with engineering experts before cracking into the building foundation, but I am sure a number of people out there have dealt with these issues, and have opinions on how to handle such things. Sure, I would like a new, custom-designed building - heck, throw in an aberration-corrected FE-TEM while you're at it - but that isn't reality. All I am looking for is one small patch (~1 m^2) of vibration-free floor. Thanks.
------------------------------------------
Phil Ahrenkiel, Assistant Professor
Nanoscience and Nanoengineering Ph.D. Program
South Dakota School of Mines and Technology
501 E. Saint Joseph St.
Rapid City, SD 57701 U.S.A.
Office: EP 221
Phone: 605-394-5238, Fax: 605-394-2365
Email: Phil.Ahrenkiel-at-sdsmt.edu



==============================Original Headers==============================
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From: allan.mitchell-at-stonebow.otago.ac.nz
Date: Thu, 29 Jul 2010 22:19:51 -0500
Subject: [Microscopy] Immuno-labeling Lowicryl HM20 sections

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

Hi All

In the ongoing quest to improve our immuno-results , and learn more
about working with Lowicryl HM20, we have come across a number of
papers recently where people have pretreated their Lowicryl HM20
sections before immuno-labeling with saturated NaOH in absolute
ethanol, for 2 to 3 seconds. This is followed by a wash then
preincubation in 0.1% sodium borohydride. This does seem like quite
harsh treatment, and I am curious as to the purpose of the saturated
NaOH step, and its origins. Have any of you come across this
before? Has anybody got the original reference?

Many thanks for your time, I hope you all have a great weekend

Allan


Allan Mitchell
Microscopy Otago - Electron Microscopy
c/- Department of Anatomy and Structural Biology
Otago School of Medical Sciences
P.O. Box 913
Dunedin
New Zealand

Phone (03) 479 5642 or 479 7301
Fax (03) 479 5086 or 479 7254

EM Centre: http://ocem.otago.ac.nz/
Confocal Centre: http://occm.otago.ac.nz/
NZ Microscopy Society: http://microscopynz.co.nz/




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From: bengu-at-fen.bilkent.edu.tr
Date: Thu, 29 Jul 2010 23:56:21 -0500
Subject: [Microscopy] Problem with VPSE G3 detector

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


Dear All,

We are having problems with a VPSE G3 type low vacuum detector on our C-Z
EVO 40 SEM.

When the detector is turned on (EP mode, Dry or Wet) we get periodic white
horizantal stripes on the screen. These resemble charging stripes but
actually are very uniform in shape and position. The image quality is very
poor actually can only "see" something at 40-100X, that is all. The
bias/gain does not work on the detector tab. We can get of the stripes if
we increase the pressure above 300 Pa, but still image quality is bad
enough.

EMServer log has this constant message "VPSE Bias = 1350". We have went
back and checked logs from previous succesful sessions with VPSE and did
not encounter any such message...

In HV we can turn on SE and BSE, so we think the issue is isolated to the
VPSE detector.

Any ideas would help a lot !!!

Best

Erman Bengu



=================================
Erman Bengu

Assistant Professor of Chemistry
Department of Chemistry
Bilkent University

Mailing Address:
Bilkent University,
Department of Chemistry,
06800, Bilkent, Ankara
Turkey

Office: SB #311
E-mail: bengu_AT_fen.bilkent.edu.tr
Phone (Office): +90 (312) 290-2153
(Lab1): +90 (312) 290-2663
(Lab2): +90 (312) 290-3332
Fax: +90 (312) 266-4068
Web: http://www.fen.bilkent.edu.tr/~bengu
==================================


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From: beaurega-at-westol.com
Date: Fri, 30 Jul 2010 00:02:31 -0500
Subject: [Microscopy] TEM reusing OsO4

Contents Retrieved from Microscopy Listserver Archives
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 Josif,

It is possible to reoxidize OsO2 or osmium chloride back up to OsO4.

Years ago, the late Dr. Chuck Garber told me it was "impossible". I read an item on the Internet from the Asian subcontinent that said essentially that an unopened bottle of OsO2 powder had a black Os stained PE bottle cap liner. So it appeared to me that he was wrong and the cap was slowly stained with OsO4 over a few years.

In my research, I found that the reaction will proceed but it is extremely slow with even strong oxidizers. There is some rate determining step that is slowing the reaction down, IMO. I had no proof back in 2001 other than a very slow reaction rate. Is this practical? Not really and I surely tried all sorts of common oxidizers.

As for using hydrogen peroxide with Os, I wouldn't! For me, osmium dioxide catalyzed the decomposition of 30% H2O2 as a very violet foaming reaction back in 2001. That's just a little heads up safety tip. Mixing Os salts and the oxidizer H2O2 is a not a good idea.

HTH,

Paul Beauregard


At 02:05 PM 7/29/10 -0500, rcmoretz-at-gmail.com wrote:
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From: leunissen-at-aurion.nl
Date: Fri, 30 Jul 2010 00:37:11 -0500
Subject: [Microscopy] Re: TEM reusing OsO4

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

Hello Josif,

I agree with Paul: because of the redox potential for the OsO4 {} OsO2 reaction a very strong oxidizing agent would have to be used to regenerate OsO4 from reduced solutions (used fixative). Even if this were to work, this would require serious safety precautions. Secondly, the mix would have to be purified after the regeneration.
However, there is a procedure to recover "unused" OsO4 from used fixative by distillation. I have done this on a regular basis in the grey past, would have to search for the detailed procedure. Let me know if this would be of interest, I could have a go at it. Before embarking on such an adventure, please be aware that such distillations need to be well-controlled as well because of the chemical hazard posed by OsO4, especially in the vapor phase.

Cheers

Jan Leunissen
AURION


On 30/07/2010, at 5:02 PM, beaurega-at-westol.com wrote:

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} Josif,
}
} It is possible to reoxidize OsO2 or osmium chloride back up to OsO4.
}
} Years ago, the late Dr. Chuck Garber told me it was "impossible". I read an item on the Internet from the Asian subcontinent that said essentially that an unopened bottle of OsO2 powder had a black Os stained PE bottle cap liner. So it appeared to me that he was wrong and the cap was slowly stained with OsO4 over a few years.
}
} In my research, I found that the reaction will proceed but it is extremely slow with even strong oxidizers. There is some rate determining step that is slowing the reaction down, IMO. I had no proof back in 2001 other than a very slow reaction rate. Is this practical? Not really and I surely tried all sorts of common oxidizers.
}
} As for using hydrogen peroxide with Os, I wouldn't! For me, osmium dioxide catalyzed the decomposition of 30% H2O2 as a very violet foaming reaction back in 2001. That's just a little heads up safety tip. Mixing Os salts and the oxidizer H2O2 is a not a good idea.
}
} HTH,
}
} Paul Beauregard
}
}
} At 02:05 PM 7/29/10 -0500, rcmoretz-at-gmail.com wrote:
} }
} }
} }
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} }
} } There is a lot of old literature on this from the 1960s & 70s.
} } Unfortunately I no longer have the references as they evaporated along
} } with all of my reprints when I retired. A search of PubMed should
} } bring up some references.
} }
} } Roger Moretz, Ph.D.
} }
} } On Thu, Jul 29, 2010 at 10:23 AM, {jmircheski-at-us.es} wrote:
} } }
} } }
} } }
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} } }
} } } Hi All,
} } }
} } } I have some memory of reading somewhere or hearing from someone that the
} } } OsO4 (used for fixing TEM samples) can be reused by adding some hydrogen
} } } peroxide. Could someone tell me if this is true and if anyone has experience
} } } in reusing the OsO4? It is quite valuable, so if this is true I‚d like to
} } } save as much as possible of it.
} } } Looking forward to your information!
} } }
} } } Regards,
} } }
} } } Dr. Josif Mircheski
} } } ____________________________________________________________________________
} } } ___
} } } Instituto de Biomedicina de Sevilla (IBIS)
} } } Hosp.Univ. Virgen del Rocío/CSIC/Univ. de Sevilla y
} } } Dpto. Fisiologia Médica y Biofísica
} } } Universidad de Sevilla
} } } Facultad de Medicina
} } } Avda. Sánchez-Pizjuán 4
} } } 41009-Sevilla
} } }
} } } Phone:+34-954556103
} } } Fax:+34-954551769
} } } e-mail: jmircheski-at-us.es
} } }
} } }
} } }
} } }
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From: nizets2-at-yahoo.com
Date: Fri, 30 Jul 2010 05:58:01 -0500
Subject: [Microscopy] Immuno-labeling Lowicryl HM20 sections

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

a detailled description how to re-destillate used OsO4 (" Os-waste") is
given in:
Swiss Chem *8 * (12), 43--44, 46 (1986)
However, in my opinion, the apparative and practical efforts are in no
proportion to the price you have to pay for new one.
I recall that in the 70th we got quite a amount of money for our
Os-waste from Degussa (a German company specialized in rare metals) but
nowadays in my experience no company even takes the waste back for free.

greetings,
Peter Heimann

--
====================================================

Dr. Peter Heimann Raum: W7-107 / Tel.: +49-(0)521-106-5628
Universitaet Bielefeld Fax: +49-(0)521-106-5654
"Zellbiologie / Cell Biology" W7-107
33501 Bielefeld, Germany www.uni-bielefeld.de/biologie/cellbio



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From ggoodwin-at-northlink.net Fri Jul 30 04:03:16 2010
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Hi Allan!

I am probably not the best IHC specialist of this list But here is what I
understand:

Ethanolic solutions of NaOH are used for etching, that is revealing the
antigenic sites embedded in the resin.
This is indeed a harsh treatement because it is not easy to etch the resin! For
this reason the treatment must be very brief and must be followed by thourough
washing. All treatments have a downside and clearly etching may destroy some
antigenic sites too, so for some it works and some others not.
As for borohydride, I am not sure but this is a reducing agent and it may be
used to reduce the free aldehyde groups present from the fixation with
glutaraldehyde. Apart from that it may also simply open certain bonds that can
hamper the immunological reaction.
I am sorry I have no references to offer, these infos just come from the far end
of my memory and I cannot say anymore where I found them.
I would be happy to be corrected if necessary.

Regards,

Stephane

 


----- Original Message ----
X-from: "allan.mitchell-at-stonebow.otago.ac.nz"
{allan.mitchell-at-stonebow.otago.ac.nz}
To: nizets2-at-yahoo.com
Sent: Fri, July 30, 2010 5:23:45 AM

Hi All

In the ongoing quest to improve our immuno-results , and learn more 
about working with Lowicryl HM20, we have come across a number of 
papers recently where people have pretreated their Lowicryl HM20 
sections before immuno-labeling with saturated NaOH in absolute 
ethanol, for 2 to 3 seconds.  This is followed by a wash then 
preincubation in 0.1% sodium borohydride.  This does seem like quite 
harsh treatment, and I am curious as to the purpose of the saturated 
NaOH step, and its origins.    Have any of you come across this 
before? Has anybody got the original reference?

Many thanks for your time, I hope you all have a great weekend

Allan


Allan Mitchell
Microscopy Otago - Electron Microscopy
c/- Department of Anatomy and Structural Biology
Otago School of Medical Sciences
P.O. Box 913
Dunedin
New Zealand

Phone  (03) 479 5642 or 479 7301
Fax    (03) 479 5086 or 479 7254

EM Centre: http://ocem.otago.ac.nz/
Confocal Centre: http://occm.otago.ac.nz/
NZ Microscopy Society: http://microscopynz.co.nz/




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From: PhillipsT-at-missouri.edu
Date: Fri, 30 Jul 2010 06:50:50 -0500
Subject: [Microscopy] Immuno-labeling Lowicryl HM20 sections

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

Sodium borohydride is usually used to reduce autofluorescence. I have had modest success with it in the distant past.

I have etched lots of Epon sections with sodium ethoxide (NaOH saturated ethanol). I usually did this to increase staining of osmicated tissue with hematoxylin and PAS or safranin O. I have tried it with Epon for colloidal gold labeling and it may have increased the signal but it increased the noise much more. I have never tried it with the Lowicryls. I believe HM20 was originally conceived as a more hydrophobic version of K4M so theoretically shouldn't be as good for immuno-staining but I also found it as good as K4M and much easier to section. But if you are doing LM work with 0.5 um sections, I strongly recommend butyl-methylmethacrylate since you can extract all the resin with acetone. I haven't done LM immuno on sections with Lowicryl or LR White or LR Gold resins since I have discovered BMMA. It may work at the EM level but my very limited tries weren't especially promising. For EM, I usually use LR Gold but often try them all. Good luck.


Thomas E. Phillips, Ph.D.
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
Biological Sciences
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (voice)
573-882-0123 (fax)

-----Original Message-----
X-from: allan.mitchell-at-stonebow.otago.ac.nz [mailto:allan.mitchell-at-stonebow.otago.ac.nz]
Sent: Thursday, July 29, 2010 10:21 PM
To: Phillips, Thomas E.

Hi All

In the ongoing quest to improve our immuno-results , and learn more
about working with Lowicryl HM20, we have come across a number of
papers recently where people have pretreated their Lowicryl HM20
sections before immuno-labeling with saturated NaOH in absolute
ethanol, for 2 to 3 seconds. This is followed by a wash then
preincubation in 0.1% sodium borohydride. This does seem like quite
harsh treatment, and I am curious as to the purpose of the saturated
NaOH step, and its origins. Have any of you come across this
before? Has anybody got the original reference?

Many thanks for your time, I hope you all have a great weekend

Allan


Allan Mitchell
Microscopy Otago - Electron Microscopy
c/- Department of Anatomy and Structural Biology
Otago School of Medical Sciences
P.O. Box 913
Dunedin
New Zealand

Phone (03) 479 5642 or 479 7301
Fax (03) 479 5086 or 479 7254

EM Centre: http://ocem.otago.ac.nz/
Confocal Centre: http://occm.otago.ac.nz/
NZ Microscopy Society: http://microscopynz.co.nz/




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From: contact-at-ibilabs.com
Date: Fri, 30 Jul 2010 08:28:00 -0500
Subject: [Microscopy] viaWWW: xray diffraction

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From: orgomezdu-at-unal.edu.co
Date: Fri, 30 Jul 2010 12:42:36 -0500
Subject: [Microscopy] viaWWW: help needed JEOL JEM 100C series

Contents Retrieved from Microscopy Listserver Archives
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Dear Collective,

I'm trying to label some root nodules and have noted in 4 separate samples that I'm getting apparently specific labeling of what appears to be starch granules, with manageable background in other parts of the tissue. That part is fine. However in all samples when I get off the sections (LR White, UV polymerized in an icebox), I am finding very even and heavy immunogold binding to the carbon support film. No formvar, just carbon. Nickel grids. The controls incubated in buffer instead of primary antibody are clean. Any idea what's going on here?

Another, kinda related, question: I'd be interested in people's thoughts on the deep, philosophical and practical ramifications of floating grids on drops vs. full immersion of grids in drops during the labeling process. I come from a long line of floaters, but some of my best friends are immersers. Is there a true way or is it all good?

Thanks and have a great weekend. Rainy and hot here. Hard to tell where the water leaves off and the air begins.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
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Fax: (573) 884-2227
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On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
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Message: We are looking for vacuum pumps for JEOL
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for the same model. Ours is serial EM1 56008-95.
With no vacuum box, no air compressor, no
manuals, no blueprints whereas mechanical, nor
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From: blai-at-aps.anl.gov
Date: Fri, 30 Jul 2010 13:29:27 -0500
Subject: [Microscopy] viaWWW: Postdoc openings

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Email: blai-at-aps.anl.gov
Name: Barry Lai

Organization: Argonne National Laboratory

Title-Subject: [Filtered] Postdoc openings

Message: The Advanced Photon Source (APS) at
Argonne National Laboratory is the USAís premier
facility for hard x-ray research. The x-ray
microscopy group at the APS is seeking highly
qualified applicants for two different
postdoctoral research associate positions aimed
at pioneering the use of cryo methods for high
resolution microscopy and trace element analysis.
An instrumentation-focused cryo-microscopy
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positions are to explore new scientific
opportunities, rather than to support user
operations at the beamlines. Interactions
(including some teaching opportunities if
desired) with Northwestern University can be
arranged.

Argonne is a U.S. Department of Energy laboratory
managed by UChicago Argonne, LLC. Argonne`s site
is approximately 25 miles southwest of Chicago on
a beautiful 1500-acre campus-like environment.
Argonne is an equal opportunity employer and we
value diversity in our workforce. Interested
candidates should send a detailed CV, list of
publications, three references, abstracts and
significant presentations through the Argonne
website at http://www.anl.gov/jobs job search for
the following requisitions:

XSD 316237 (for technical information contact Dr.
Barry Lai at blai-at-aps.anl.gov)
XSD 316253 (for technical information contact Dr.
Lydia Finney at lfinney-at-aps.anl.gov)


Barry Lai
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From: leunissen-at-aurion.nl
Date: Fri, 30 Jul 2010 13:49:57 -0500
Subject: [Microscopy] Re: Immuno TEM: Gold binding to C film

Contents Retrieved from Microscopy Listserver Archives
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Hi Randy,

X-from one floater to another...
if I understand correctly, there is specific and 'clean' labeling on the sections, but in areas next to the sections there is a persistent background.
Seems your carbon films are fond of your primaries, more fond than your sections are. Do you use a blocking step? You may have to adjust that. Does it happen with that particular primary only?

Re floaters and immersers.... Prof. Moise Bendayan in the 80's used sections on bare grids (no film). That offers the possibility of both sides of sections to become labeled when immersing grids and he used this (albeit floating) for double labeling experiments even with two primaries from the same species. When filmed grids are used there is no advantage, as long as antibodies can reach the section surface equally well between floating and immersing.
Theoretically, immersing may achieve homogeneous exposure more easily, especially with short incubations as the immunoreagent droplets are more or less stirred when grids are inserted mixing adhering liquid with reagent. This is is mimicked in the Leica IGL where grids are moved up and down on the droplet without breaking contact. Nickel grids can be made to rotate on the incubation droplet when they are incubated on a drop of Parafilm on a magnetic stirrer. Works very well, but please do not curse me when the grids stick to your tweezers or when developing an astigmatism related migraine.

A disadvantage of immersing is that on the 'hollow' side of the grid reagents may be hard to remove next to the grid bars, so increasing background.. you know how the interesting parts in your sections are always so close to or preferably even on the grid bar.....

Cheers

Jan

AURION
Honorary Fellow - University of Otago
New Zealand


On 31/07/2010, at 1:41 AM, TindallR-at-missouri.edu wrote:

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} I'm trying to label some root nodules and have noted in 4 separate samples that I'm getting apparently specific labeling of what appears to be starch granules, with manageable background in other parts of the tissue. That part is fine. However in all samples when I get off the sections (LR White, UV polymerized in an icebox), I am finding very even and heavy immunogold binding to the carbon support film. No formvar, just carbon. Nickel grids. The controls incubated in buffer instead of primary antibody are clean. Any idea what's going on here?
}
} Another, kinda related, question: I'd be interested in people's thoughts on the deep, philosophical and practical ramifications of floating grids on drops vs. full immersion of grids in drops during the labeling process. I come from a long line of floaters, but some of my best friends are immersers. Is there a true way or is it all good?
}
} Thanks and have a great weekend. Rainy and hot here. Hard to tell where the water leaves off and the air begins.
}
} Cheers,
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
}
}
} ==============================Original Headers==============================
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} 8, 30 -- Subject: Immuno TEM: Gold binding to C film
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From: PhillipsT-at-missouri.edu
Date: Fri, 30 Jul 2010 15:39:28 -0500
Subject: [Microscopy] Immuno TEM: Gold binding to C film

Contents Retrieved from Microscopy Listserver Archives
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Randy - When I was doing immunocytochemistry on LRG thin sections I found a significant advantage of immersing over floating. I don't know if it was 2x as much labeling but maybe.

Have you tried acetylated BSA or cold water fish gelatin in your blocking buffer? I also like to add 1% normal serum from whatever animal the secondary was made in. I am a big fan of long incubations for primary and secondary staining and often use 4 hrs to block before starting overnight incubation in the primary. Unrelated to the problem you asked about but often with polyclonal primaries on plant material, it pays to pre-absorb the primary on control tissue extract if the antigen is a transgenic expressed protein or some bacterium not present on normal nodules.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/



On 31/07/2010, at 1:41 AM, TindallR-at-missouri.edu wrote:

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} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
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} Dear Collective,
}
} I'm trying to label some root nodules and have noted in 4 separate samples that I'm getting apparently specific labeling of what appears to be starch granules, with manageable background in other parts of the tissue. That part is fine. However in all samples when I get off the sections (LR White, UV polymerized in an icebox), I am finding very even and heavy immunogold binding to the carbon support film. No formvar, just carbon. Nickel grids. The controls incubated in buffer instead of primary antibody are clean. Any idea what's going on here?
}
} Another, kinda related, question: I'd be interested in people's thoughts on the deep, philosophical and practical ramifications of floating grids on drops vs. full immersion of grids in drops during the labeling process. I come from a long line of floaters, but some of my best friends are immersers. Is there a true way or is it all good?
}
} Thanks and have a great weekend. Rainy and hot here. Hard to tell where the water leaves off and the air begins.
}
} Cheers,
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
}
}
} ==============================Original Headers==============================
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From: wtivol-at-verizon.net
Date: Fri, 30 Jul 2010 15:40:02 -0500
Subject: [Microscopy] Re: Fwd: Ask-A-Microscopist

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On Jul 28, 2010, at 12:45 PM, oshel1pe-at-cmich.edu wrote:

} realname - Mark Twigg
} Email - twigg-at-estd.nrl.navy.mil
} EDUCATION - Graduate College
} LOCATION - Washington, DC
} SUBJECT_OF_QUESTION - HRTEM Siting Near Laser
} QUESTION - My organization is slated to be moved to a new building
} sometime in the next several years. Of course that raises the
} question concerning where to site my HRTEM. Currently I have a top
} entry Hitachi H9000UHR. By the time this building is ready (maybe
} six years in the future) we may have a new HRTEM instrument with
} more demanding requirements. The problem is that the floor plan for
} the building has just been altered with my lab positioned right next
} to an optics lab with a high-powered laser designed to blast
} electronic devices as a way of simulating radiation effects.
} Anybody with horror tales about living next door to one of these
} things?
}
} Thanks,
}
} Mark
--
Dear Mark,
The only time I've been at a TEM next to a laser, it was an ultrafast
TEM, where the laser produced the beam. It was not high resolution,
so my experience doesn't really apply. If the laser you're concerned
with is operating at present, I would suggest making the appropriate
mechanical and electrical measurements at various places near and/or
in the laser lab. I'm sure that Hitachi will have the necessary
equipment, and I expect they'd be motivated to help you. If your
measurements indicate any problems, you do seem to have significant
lead time to work out solutions. Good luck.
Yours,
Bill


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From: rbeavers-at-mail.smu.edu
Date: Fri, 30 Jul 2010 16:23:52 -0500
Subject: [Microscopy] EDAX data export

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Group

Does anyone know if you are able to export raw EDS data from a EDAX system.

Would like to export raw energy vs counts spectrum data to a program one of our faculty has written to test its accuracy.

Thanks

Roy Beavers
Southern Methodist University
Department of Earth Sciences
P.O. Box 750395
Dallas, TX  75275
Voice: 214-768-2756
Fax: 214-768-2701
Email: rbeavers-at-smu.edu



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From: gary-at-gaugler.com
Date: Fri, 30 Jul 2010 17:04:00 -0500
Subject: [Microscopy] Re: EDAX data export

Contents Retrieved from Microscopy Listserver Archives
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Raw data files for EDAX is their .spc file. They provide
a utility called "EDAX Spectral Processing Utility" which
has a feature for SPC} MSA. I have not been able to get
this to work but I have had no reason to export. I work
with their .spc and .spd files and that is about it, other
than .tif image files.

There is probably some other way to do this or perhaps I
am simply doing something wrong. This helper app is under
Genesis 5.2 and earlier versions of Genesis.

Hope this helps.

gary g.

At 02:25 PM 7/30/2010, you wrote:



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From: ejb1176-at-gmail.com
Date: Tue, 3 Aug 2010 16:12:26 -0500
Subject: [Microscopy] DB235 ion pump issues - results...

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Hi, Randy:

I think this type of non-specific “sticking” may have something to do with the static or hydrophobic nature of carbon film. What happened might be that when one did not block or did not block sufficiently before primary incubation, primary would stick to the carbon film (by hydrophobic interaction for example) and in turn be bound by the secondary. In the case of the control grid, if one used the same solution minus primary, the carbon surface could be further blocked by the protein additives in the incubation buffer and in the subsequential rinse buffer, which would help prevent the sticking of the secondary.

Do you treat your grids by glow discharge before putting sections on? I have never compared the results with and without glow discharge, but would like to know if you want to try.

Thank you. See you in Portland?

Hong
________________________________________
X-from: TindallR-at-missouri.edu [TindallR-at-missouri.edu]
Sent: Friday, July 30, 2010 9:42 AM
To: Yi, Hong

Dear Collective,

I'm trying to label some root nodules and have noted in 4 separate samples that I'm getting apparently specific labeling of what appears to be starch granules, with manageable background in other parts of the tissue. That part is fine. However in all samples when I get off the sections (LR White, UV polymerized in an icebox), I am finding very even and heavy immunogold binding to the carbon support film. No formvar, just carbon. Nickel grids. The controls incubated in buffer instead of primary antibody are clean. Any idea what's going on here?

Another, kinda related, question: I'd be interested in people's thoughts on the deep, philosophical and practical ramifications of floating grids on drops vs. full immersion of grids in drops during the labeling process. I come from a long line of floaters, but some of my best friends are immersers. Is there a true way or is it all good?

Thanks and have a great weekend. Rainy and hot here. Hard to tell where the water leaves off and the air begins.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From ggins-at-insur-e.net Sat Jul 31 00:51:08 2010
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EDAX software has a "save as" function "Spectrum as text" that saves the
data as two column text of counts v. energy. The extension is ".csv". Which
can be opened in any spreadsheet.

Roger

Roger A. Ristau, PhD
Electron Microscopy Specialist
Institute of Materials Science
97 North Eagleville Road
University of Connecticut
Storrs, CT 06269
vox: 860-486-5453
fax: 860-486-4745


} From: {rbeavers-at-mail.smu.edu}
} Reply-To: {rbeavers-at-mail.smu.edu}
} Date: Fri, 30 Jul 2010 16:26:57 -0500
} To: {raristau-at-ims.uconn.edu}
} Subject: [Microscopy] EDAX data export
}
}
}
}
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} Does anyone know if you are able to export raw EDS data from a EDAX system.
}
} Would like to export raw energy vs counts spectrum data to a program one of
} our faculty has written to test its accuracy.
}
} Thanks
}
} Roy Beavers
} Southern Methodist University
} Department of Earth Sciences
} P.O. Box 750395
} Dallas, TX  75275
} Voice: 214-768-2756
} Fax: 214-768-2701
} Email: rbeavers-at-smu.edu
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Dear Phil,

We've had a lot of discussion with Michael Gendreau of Colin Gordon &
Associates about the merits (& demerits) of isolation slabs, i.e. small
patches of floor that are mechanically 'decoupled' by use of some medium
with a very different mechanical impedance e.g. elastomer, air, etc .
However, I know our circumstances are different (new building vs already
existing building), but there are a few simple principles that Michael's
analysis demonstrated beautifully (using data taken from our already
existing isolation slabs).

An isolated slab is, in itself, a resonator with resonant frequencies
determined by the density and elasticity of the material as well as the
spatial extent (modes of oscillation). Generally, the bigger and heavier
the mass, the lower the resonant frequency: a large single mass is better
than lots of little isolated slabs if you need the resonant frequency
shifted downwards.

Isolated slabs are pretty ineffectual below their resonant frequencies (the
wavelength of the vibrations are much longer the slab dimensions and the
slab moves easily), they amplify the vibrations in the vicinity of the
resonant frequency* before giving attenuation with approximately a 1/f
dependence for high frequencies.

Given the 18 Hz oscillation you have, you'd probably need a very large and
deep mass to get a resonant frequency low enough to get some attenuation,
probably something much larger than the 1 by 1 m^2 you're suggesting.

If the noise is a relatively sharp tone (single frequency), the active
noise cancelling system would probably work. If the noise is spread over a
wide band of frequencies, it is going to be much harder to remove.

You said that source appears to be HVAC. Could the air-ducts be
retro-fitted with noise-damping baffles? Or the duct-work re-configured to
remove most of the flow away from your workspace?

Yours, Jon Barnard

* In our case we had amplification factors of between 3 and 10 times for
different microscope/slab systems.

P.S. I have no financial interest with Colin Gordon & Associates.

}
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}
} We will be installing a new 200-KV LaB6 TEM within the next few months,
} and our university has allocated a nice lab in the basement of our older
} (ca. 1972) building for the instrument. But the environmental survey
} showed an on-going, 18-Hz, 25-um vertical oscillation in that room due to
} the building's HVAC. Also, the floor lacks resiliency in some areas,
} maybe due to shifting over time of the soil below, so occasional, nearby
} foot traffic could affect image stability.
}
} The survey company offers an active vibration-cancellation system to
} remove the low-frequency noise. Though I am not opposed to this, my first
} priority is to address any issues with the building foundation before
} installation. So I have spoken with several contractors about the
} possibility of forming an isolated region of the floor to support the
} TEM.
}
} My understanding is that the vibration propagates readily through the
} rigid cement floor, which lacks sufficient mass to damp the oscillation.
} It would seem that the ideal floor has 1) large mass and 2) weak coupling
} to the driving force.
}
} One practical possibility I investigated is to install a detached,
} separate section of floor on concrete piers, set deep (12-20 ft) into the
} ground, upon which the TEM column would rest. Now, I am not a
} civil/mechanical engineer, but my na�ve impression is that if the piers
} extend deep enough, the earth would essentially act as a massive load.
} The underlying soil is relatively soft, so the vibrations from the
} building would not substantially propagate through the earth from the
} building to the TEM. Plus, this should decouple the column from the
} higher-frequency traffic in the building.
}
} The employee of the microscope manufacturer with whom I spoke was adamant
} that this was a ridiculous idea, and would actually make the vibration
} worse. He believes it would require an impractically massive foundation
} to impede these oscillations, that the earth would not damp the
} vibration, and that the active cancellation system is our only option.
}
} Of course, we will consult with engineering experts before cracking into
} the building foundation, but I am sure a number of people out there have
} dealt with these issues, and have opinions on how to handle such things.
} Sure, I would like a new, custom-designed building - heck, throw in an
} aberration-corrected FE-TEM while you're at it - but that isn't reality.
} All I am looking for is one small patch (~1 m^2) of vibration-free floor.
} Thanks.
} ------------------------------------------
} Phil Ahrenkiel,
} Assistant Professor Nanoscience and Nanoengineering Ph.D. Program South
} Dakota School of Mines and Technology 501 E. Saint Joseph St. Rapid City,
} SD 57701 U.S.A. Office: EP 221 Phone: 605-394-5238, Fax: 605-394-2365
} Email: Phil.Ahrenkiel-at-sdsmt.edu



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13, 30 -- From jsb43-at-hermes.cam.ac.uk Tue Aug 3 08:15:01 2010
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From anmeldung-at-gebirgsverein.at Tue Aug 3 13:48:22 2010
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Received: from [87.218.250.152] by mxlb.ispgateway.de; Tue, 3 Aug 2010 19:46:03 +0100

Many thanks to all who responded to our ion pump dilemma.

With ListServe respondents' advice the problem was eventually tracked
down to a bad cold cathode Penning gauge.
{e-7 mbar is not a valid reading on this instrument.

Once the gauge was replaced and the system allowed to pump overnight,
a valid vacuum reading was obtained.
It is noteworthy that the vacuum readout was still {e-7 mbar even with
the CCG disconnected, indicating CCG failure.

The column isolation valve was able to be opened under the service
login....in order to bring the ion column vacuum down to the e-7 mbar
range.
The ion pump re-started with no arcing....
It took some effort to get the LMIS to fire and stay on, but
eventually it was coaxed to stability.

We are back in operation with only having to re-align the ion column apertures.

Thanks again to all who sent suggestions.

cheers
ed


--
Edward Basgall
103 N. Drexel Ave.
Havertown, PA 19083

c: 610-574-4007

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From: rowland3-at-umbc.edu
Date: Thu, 5 Aug 2010 00:55:19 -0500
Subject: [Microscopy] viaWWW: Analog to Digital conversion for Jeol jsm-6100

Contents Retrieved from Microscopy Listserver Archives
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Hi everybody!

We have in our lab an ULVAC ionization gauge controller, GI-N3.
We recently aquired another vacuummeter, and it shows almost double of the
pressure measured with the ionization gauge.

Until now we have set the emission of the ionization gauge to 5 when
the emission check
button was depressed. But now I wonder if it should set to 10 the same
way, as it reads then
almost the same pressure as the other gauge.

Does anybody have a user manual for this type of controller? If so,
could provide me the
necessary details from it?

Thanks

Jakab-Farkas Laszlo

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From anna-at-hive.dk Wed Aug 4 23:07:06 2010
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This Question/Comment was submitted to the Microscopy Listserver
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Email: rowland3-at-umbc.edu
Name: Cody Rowland

Organization: Matsys

Title-Subject: [Filtered] Analog to Digital conversion for Jeol jsm-6100

Message: Hello I'm an SEM newbie.

Attempting to revive an abandoned jsm-6100.
To date I've connected a roughing pump, nitrogen supply, and water
cooling. I used JEOL circuit diagrams to aid me in reconnection of
the original cabling. This SEM was originally fitted with a diffusion
pump and at some stage had been switched over to a Pfeiffer TPH240
turbo pump.

Currently, I'm in the process of trouble shooting the turbo pump
controller. But, when powering up the SEM the diaplay and all other
non-vacuum functions appear to be working.

At some point in the future I will be interested in converting the
Analog signal to a digital, so I can control the SEM more easily
using some common software to handle the digital image. Does anyone
have any suggestions on where to look or if they have done a similar
conversion what kind of things I should watch out for.

Thanks
Cody

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From: ldemp-at-mse.ufl.edu
Date: Thu, 5 Aug 2010 08:18:23 -0500
Subject: [Microscopy] viaWWW: On-line Materials Characterization Certificate

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

This Question/Comment was submitted to the Microscopy Listserver
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Email: ldemp-at-mse.ufl.edu
Name: Luisa Amelia Dempere

Organization: University of Florida

Title-Subject: [Filtered] On-line Materials Characterization Certificate

Message: Registration is open for the courses "Survey of Materials
Characterization Techniques" and "Transmission Electron Microscopy"
offered this fall 2010. Both courses count towards the on-line
graduate certificate in Materials Characterization offered by the
Materials Science and Engineering Department at the University of
Florida through UF EDGE www.ufedge.ufl.edu



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From: ibarke2-at-uwo.ca
Date: Thu, 5 Aug 2010 15:55:43 -0500
Subject: [Microscopy]

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



Hello,

We have an older Hitachi S2500 Tungsten SEM that's hooked up to an old Windows98 computer,C with two ISA-slot control boards. Recently the computer's hard drive failed and we had to replace it. However, we do not have the original Quartz PCI program (I believe it's version 6), or the drivers
that are for the ISA control boards that are inside the computer. We have
contacted Hitachi, but they do not seem to have the original software that's compatible with our setup. The reason we do not upgrade the computer,
from my understanding, is that the ISA boards are not compatible with
Windows XP therefore it would be an expensive upgrade to upgrade the boards to PCI-slotted.
Does anyone have an older version of Quartz PCI and the control board drive
rs that are compatible with Windows 98 and the ISA boards that they could send us?
Please send me an email back at: ibarke2-at-uwo.ca
Thanks for your time.


Ivan R. Barker
UWO Earth Sciences MSc
ZAPLab Geochronology Research Technician
T: 519-661-2111 ext. 88397
E: ibarke2-at-uwo.ca






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From: ibarke2-at-uwo.ca
Date: Fri, 6 Aug 2010 08:50:59 -0500
Subject: [Microscopy] SEM - Need Quartz PCI & Drivers for Hitachi S2500 - Win98 & ISA

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Hello,

Sorry for the duplicate email, I sent this email yesterday, but forgot a subject.
We have an older Hitachi S2500 Tungsten SEM that's hooked up to an old Windows98 computer, with two ISA-slot control boards. Recently the computer's hard drive failed and we had to replace it. However, we do not have the original Quartz PCI program (I believe it's version 6), or the drivers that are for the ISA control boards that are inside the computer. We have contacted Hitachi, but they do not seem to have the original software that's compatible with our setup. The reason we do not upgrade the computer,from my understanding, is that the ISA boards are not compatible with Windows XP therefore it would be an expensive upgrade to upgrade the boards to PCI-slotted.Does anyone have an older version of Quartz PCI and the control board drivers that are compatible with Windows 98 and the ISA boards that they could send us?
Please send me an email back at: ibarke2-at-uwo.ca
Thanks for your time.


Ivan R. Barker
UWO Earth Sciences MSc
ZAPLab Geochronology Research Technician
T: 519-661-2111 ext. 88397
E: ibarke2-at-uwo.ca
 





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From: mshaw1-at-lumc.edu
Date: Fri, 6 Aug 2010 12:13:35 -0500
Subject: [Microscopy] viaWWW: colocalization software

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Email: mshaw1-at-lumc.edu
Name: Molly Shaw

Organization: Loyola University Chicago

Title-Subject: [Filtered] colocalization software

Message: Hi there,

My lab is looking to purchase colocalization software, so I was
wondering if any of you have recommendations. We are using a Zeiss
LSM 510, so we don't need imaging capabilities, just colocalization
quantification.

We have been using ImageJ, but it doesn't seem to be able to do
everything we need. Our main issue is that we're trying to colocalize
a protein with a non-protein sphere. The protein does not actually
enter the sphere, but we know they do associate. However, this
association is not being reflected in the coefficients generated by
ImageJ (Pearson is only about 0.3). We're wondering if it's necessary
to use a program that could treat our protein as a sphere (or
something similar) and thus quantify colocalization more accurately.

Any suggestions are welcome.

Thank you,
Molly

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From: kenconverse-at-qualityimages.biz
Date: Fri, 6 Aug 2010 14:31:04 -0500
Subject: [Microscopy] viaWWW: Analog to Digital conversion for Jeol jsm-6100

Contents Retrieved from Microscopy Listserver Archives
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Cody,
Are you planning on having any EDS capability? That can include digital
imaging either as standard or as an option.

There are a number of systems out there. I'm not going to even try to hit
them all, but 4 Pi, Orion, Quartz PCI are possibilities. Dale Callaham at
UMass Amherst has Image Thief on line and may come up with a USB version
http://people.umass.edu/dac/projects/ImageThief/ImageThief.html

If you're good at programming, Data Translation and National Instruments
have USB modules for analog and digital I/O so you can make your own passive
or active capture system.

I'm not familiar with the JSM-6100, but the 6400 and 6300 each have a
digital screen (right side) that could be tapped into for lower resolution
images.

There's lots of info out there.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


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Email: rowland3-at-umbc.edu
Name: Cody Rowland

Organization: Matsys

Title-Subject: [Filtered] Analog to Digital conversion for Jeol jsm-6100

Message: Hello I'm an SEM newbie.

Attempting to revive an abandoned jsm-6100.
To date I've connected a roughing pump, nitrogen supply, and water
cooling. I used JEOL circuit diagrams to aid me in reconnection of
the original cabling. This SEM was originally fitted with a diffusion
pump and at some stage had been switched over to a Pfeiffer TPH240
turbo pump.

Currently, I'm in the process of trouble shooting the turbo pump
controller. But, when powering up the SEM the diaplay and all other
non-vacuum functions appear to be working.

At some point in the future I will be interested in converting the
Analog signal to a digital, so I can control the SEM more easily
using some common software to handle the digital image. Does anyone
have any suggestions on where to look or if they have done a similar
conversion what kind of things I should watch out for.

Thanks
Cody

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From: jmircheski-at-us.es
Date: Fri, 6 Aug 2010 17:38:24 -0500
Subject: [Microscopy] Re: viaWWW: colocalization software

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Dear Molly,

I am not an expert in analysing colocalization, nor I have much experience in it, but I know that there different ways of doing it, depending on what do you need. There is one version of ImageJ, called MacBiophotonics ImageJ with integrated sets of different plugins. I think that it has more than one option of analysing colocalization. The website of MBF ImageJ has some good explanation of almost all of the plugins, how they work and how to set them (http://www.macbiophotonics.ca/imagej/). In addition, there should be some additional plugins on the ImageJ website. I know that I got different results once when I tried to analyse one of my images with different plugins (which ones I used I don't know).
Hope this helps.

Regards,

Josif Mircheski, MD

Dpt. of Physiology
University of Seville, Spain

----- Mensaje original -----
De: mshaw1-at-lumc.edu
Fecha: Viernes, Agosto 6, 2010 7:24 pm
Asunto: [Microscopy] viaWWW: colocalization software
A: jmircheski-at-us.es

}
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} Email: mshaw1-at-lumc.edu
} Name: Molly Shaw
}
} Organization: Loyola University Chicago
}
} Title-Subject: [Filtered] colocalization software
}
} Message: Hi there,
}
} My lab is looking to purchase colocalization software, so I was
} wondering if any of you have recommendations. We are using a
} Zeiss
} LSM 510, so we don't need imaging capabilities, just
} colocalization
} quantification.
}
} We have been using ImageJ, but it doesn't seem to be able to do
} everything we need. Our main issue is that we're trying to
} colocalize
} a protein with a non-protein sphere. The protein does not
} actually
} enter the sphere, but we know they do associate. However, this
} association is not being reflected in the coefficients generated
} by
} ImageJ (Pearson is only about 0.3). We're wondering if it's
} necessary
} to use a program that could treat our protein as a sphere (or
} something similar) and thus quantify colocalization more accurately.
}
} Any suggestions are welcome.
}
} Thank you,
} Molly
}
}   Login Host: 147.126.254.237
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From: cpawlowicz-at-ubmtechinsights.com
Date: Mon, 9 Aug 2010 07:45:19 -0500
Subject: [Microscopy] RE: small valves

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Email: katev-at-uwm.edu
Name: Kathryn Valerius

Organization: Univ. of Wisconsin-Milwaukee, Dept. of Physics

Title-Subject: [Filtered] Posting a Job
Announcement for Senior Research Scientist

Message: Kindly share/post the following for a
position for a Senior Research Scientist:

THE UNIVERSITY OF WISCONSIN ñ MILWAUKEE

Senior Research Scientist

The Department of Physics at the University of
Wisconsin ñ Milwaukee seeks outstanding
applicants for a Senior Scientist position for
the AY 2010/11, at a 70% appointment. Candidates
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Responsibilities will include: conducting
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data analysis of current and new research
projects; performing experimental work in HRTEM
lab in support of ongoing research; providing
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Marija Gajdardziska-Josifovska via email at
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From apatel15-at-utk.edu Sat Aug 7 10:36:22 2010
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I need some really small ball or plug valves for 1/8" or 1/4" tubing
for a gadget I'm designing. Does anyone know of a convenient source
for such items? I've done a bit of searching on the Internet, and
found some that appear to be standard size, but nothing small enough
to be appealing.

Wil Bigelow
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From andreas.turwitt-at-tua-electronics.com Sat Aug 7 20:39:47 2010
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We use a bunch of different 1/8 and 1/4 valves from McMaster-Carr.. nitrogen lines, compressed air, vacuum etc.

http://www.mcmaster.com/#ball-valves

Chris Pawlowicz, B.Eng
Manager, Lab Development
UBM TechInsights
3000 Solandt Rd Ottawa ON Canada K2K 2X2
T: +1 613 599 5145 x4454
F: +1 613 599 6501
E: cpawlowicz-at-ubmtechinsights.com

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I need some really small ball or plug valves for 1/8" or 1/4" tubing for a gadget I'm designing. Does anyone know of a convenient source for such items? I've done a bit of searching on the Internet, and found some that appear to be standard size, but nothing small enough to be appealing.

Wil Bigelow
--

CONFIDENTIALITY NOTICE: This e-mail transmission, and any documents, files or previous e-mail messages attached to it may contain confidential information that is legally privileged. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any disclosure, copying, distribution or use of any of the information contained in or attached to this transmission is STRICTLY PROHIBITED. If you have received this transmission in error, please immediately notify the sender. Please destroy the original transmission and its attachments without reading or saving in any manner. Thank you in advance for your cooperation.


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From: kenconverse-at-qualityimages.biz
Date: Mon, 9 Aug 2010 08:10:27 -0500
Subject: [Microscopy] small valves

Contents Retrieved from Microscopy Listserver Archives
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Will,
Are you looking for standard pipe thread or something smaller? I found this
http://www.specialtymfg.com/ball_valves_brass/default.asp
that uses 10-32 gasketed fittings (Clippard type). Maybe they are small
enough.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: bigelow-at-umich.edu [mailto:bigelow-at-umich.edu]
Sent: Saturday, August 07, 2010 5:39 PM
To: kenconverse-at-qualityimages.biz

I need some really small ball or plug valves for 1/8" or 1/4" tubing
for a gadget I'm designing. Does anyone know of a convenient source
for such items? I've done a bit of searching on the Internet, and
found some that appear to be standard size, but nothing small enough
to be appealing.

Wil Bigelow
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From: Kylie.Brown2-at-jcu.edu.au
Date: Mon, 9 Aug 2010 08:30:23 -0500
Subject: [Microscopy] viaWWW: Problem with JEOL JSM-6300 Emission Current Light

Contents Retrieved from Microscopy Listserver Archives
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Email: Kylie.Brown2-at-jcu.edu.au
Name: Kylie

Organization: James Cook University- AUSTRALIA

Title-Subject: [Filtered] Problem with JEOL JSM-6300 Emission Current Light

Message: I am currently experiencing problems with our JEOL JSM-6300.
Let me first say that I have a love for our old clunker of a machine
and I am the only one that uses it to take photographs/help others
photograph/research, however I am not an equipped technician when it
comes to the electronics of the machine and I think I have bitten off
more than I can chew. (insert very stressed face here)

I have just had a technician (from moran scientific) to look at the
SEM and service it (april 2010) and it was working fine up until 2
weeks ago.
My emission current filament light is not "lighting up" however if I
push the FILA button there is current and there is a picture on the
screens. This picture however is just vertical stripes (not the
katydid wings i need to photograph). This indicates to me that there
is a scanning problem???
So....
I have checked the RP, DP, Water and that is all fine
There is no error LED's on the vacuum display (so chamber is at vacuum)
I have also checked the fuses at the back of the console (the 13
that are visible).
The technician from Moran told me that there are another 40 or so
fuses to check but I cannot located them without taking panels off
the machine.
Does anyone know where the fuses might be located?
JEOL once did a service on the machine and indicated replacing a
certain fuse with a higher volume as the current going through it was
too high...
Also what could the possibilities be why the emission's current light
itself is not working.

Any advice/ideas would be greatly appreciated.
Many Thanks
Kylie



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From: kenconverse-at-qualityimages.biz
Date: Mon, 9 Aug 2010 09:38:21 -0500
Subject: [Microscopy] viaWWW: Problem with JEOL JSM-6300 Emission Current

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

Kylie,
The emission/filament meter may just have burnt out bulbs. There are a pair
of 28V, 35mA lamps. There may also be a broken wire going to the lamps.
There is also the possibility of a bad opto-isolator on the HT I/O PB (sheet
2 upper left). This is inconvenient, but not a show-stopper and has nothing
to do with the missing scan.

You have no vertical (Y) scan to the column. There are many possibilities
but if you shut everything down and pull the wall plug or the wall
disconnect box, then you can take the panels off the power unit (if you need
to) and check for F36, F37, F38 and F39. They are 5A fuses. (If these
fuses are visible without removing the panels, then just turn off the main
breaker and remove the Plexiglass covers to change any that are blown.)

If none of them are blown, then you probably need an oscilloscope to be able
to check the mag control board and the water-cooled mag power amp. This is
getting much deeper into the system.

Good luck!

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
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Sent: Monday, August 09, 2010 9:33 AM
To: kenconverse-at-qualityimages.biz

This Question/Comment was submitted to the Microscopy Listserver
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Email: Kylie.Brown2-at-jcu.edu.au
Name: Kylie

Organization: James Cook University- AUSTRALIA

Title-Subject: [Filtered] Problem with JEOL JSM-6300 Emission Current Light

Message: I am currently experiencing problems with our JEOL JSM-6300.
Let me first say that I have a love for our old clunker of a machine
and I am the only one that uses it to take photographs/help others
photograph/research, however I am not an equipped technician when it
comes to the electronics of the machine and I think I have bitten off
more than I can chew. (insert very stressed face here)

I have just had a technician (from moran scientific) to look at the
SEM and service it (april 2010) and it was working fine up until 2
weeks ago.
My emission current filament light is not "lighting up" however if I
push the FILA button there is current and there is a picture on the
screens. This picture however is just vertical stripes (not the
katydid wings i need to photograph). This indicates to me that there
is a scanning problem???
So....
I have checked the RP, DP, Water and that is all fine
There is no error LED's on the vacuum display (so chamber is at vacuum)
I have also checked the fuses at the back of the console (the 13
that are visible).
The technician from Moran told me that there are another 40 or so
fuses to check but I cannot located them without taking panels off
the machine.
Does anyone know where the fuses might be located?
JEOL once did a service on the machine and indicated replacing a
certain fuse with a higher volume as the current going through it was
too high...
Also what could the possibilities be why the emission's current light
itself is not working.

Any advice/ideas would be greatly appreciated.
Many Thanks
Kylie



Login Host: 137.219.155.111
---------------------------------------------------------------------------

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From: Artur.irani-at-yahoo.com
Date: Tue, 10 Aug 2010 08:10:38 -0500
Subject: [Microscopy] viaWWW: EPMA SX100 sample exchange

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Email: Artur.irani-at-yahoo.com
Name: kazem

Organization: IMPRC

Title-Subject: [Filtered] Q

Message: hi dear friends
I have really problems with EPMA SX100 when I want to bring out the
sample. Is there any one to help me about some technical problems of
this machine.
regards

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From: mark.riggs-at-asml.com
Date: Tue, 10 Aug 2010 08:53:16 -0500
Subject: [Microscopy] viaWWW: Hitachi S-4800 capability for 6"/150mm reticle inspection

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Email: mark.riggs-at-asml.com
Name: Mark Riggs

Organization: ASML Wilton, CT

Title-Subject: [Filtered] Hitachi S-4800 capability for 6"/150mm
reticle inspection

Message: i understand Type II stage for this instrument offers only
110mm x-y stage travel. Our immediate need is 150mm reticle
inspection/particle identification, but we like the s-4800 image
capability for other submissions. has anyone adapted this stage for
6" reticles, or is the chamber internal dimension too restrictive?

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From: oshel1pe-at-cmich.edu
Date: Tue, 10 Aug 2010 10:17:42 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist EMs for C nanotubes

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} Below is the result of your form, submitted on
} Monday, August 09, 2010 at 07:53:02 AM.
}
} realname - María Luján Ibarra
} Email - mlibarra27-at-gmail.com
} SUBJECT_OF_QUESTION - SEM/TEM
} QUESTION - Hello
} I®º searrching for a TEM and a SEm for my
} research on carbon nanotubes with very low
} budget. I found a SEM S-806 and a Zeiss EM-10,
} both used. I want to now your opinion about both
} equipment and if they can be useful for my
} investigation. Do you have the manuals or the
} specifications of the microscopes?
} thank you
}
} María Luján Ibarra
}
--
***************************************************************************************
Forwarded from "Ask a Microscopist"
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From: bnross-at-interchange.ubc.ca
Date: Tue, 10 Aug 2010 15:53:01 -0500
Subject: [Microscopy] TEM beam drift after filament change.

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

Hello fellow listserver colleagues,

I installed a new filament in our Hitachi H7600 yesterday and all seemed well after gun alignment etc. However, today it appears that the beam just wants to wander its merry way around the screen as if we were merely suggesting it stayed centered, rather than it being steadfastly forced into place by the strict electrostatic and magnetic forces the microscope imposes on it. The wandering isn't a great distance mind you, just a few cm this way or that, tending to fluctuate in a roughly diagonal path across the screen. The drift is mostly a very smooth and slow motion, but occasionally the beam will jump 5mm or so, and then continue on its lazy drift. Also, it doesn't seem to drift back and forth from one point to another, just away from center when you place it there with the beam shifts. It then goes off in one direction and stays put when it moves as far as it wants/needs to.

This causes a bit of a problem when imaging at higher mags as you might imagine. We suspect that we may have a box of questionable filaments, since we are only getting between 60 and 140 hours out of them without any obvious change in microscope operating conditions. (Gun vacuum seems good at around 4x10^-7 mbar.)
Are there any other obvious things things I can check that might be causing this? Help me pin down my wayward beam!

Thanks,
--
Bradford Ross

Microscopy Technician
BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996


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From: kenconverse-at-qualityimages.biz
Date: Tue, 10 Aug 2010 16:08:04 -0500
Subject: [Microscopy] TEM beam drift after filament change.

Contents Retrieved from Microscopy Listserver Archives
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Bradford,
I strongly suspect that you have a bit of "crud" (technical term) in your
column that is charging. It may be as simple as piece of dust or lint that
entered when you were changing the filament, or it may be something that was
in a harmless place but got moved when the gun was vented and/or repumped.

There are 2 hints in your description. The first is that the beam drifts,
you recenter it and it drifts again (in the same direction?). The second is
the occasional jump in position. Sounds like a discharge from some
nonconductive "crud".

Clean the column.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: bnross-at-interchange.ubc.ca [mailto:bnross-at-interchange.ubc.ca]
Sent: Tuesday, August 10, 2010 4:55 PM
To: kenconverse-at-qualityimages.biz

Hello fellow listserver colleagues,

I installed a new filament in our Hitachi H7600 yesterday and all seemed
well after gun alignment etc. However, today it appears that the beam just
wants to wander its merry way around the screen as if we were merely
suggesting it stayed centered, rather than it being steadfastly forced into
place by the strict electrostatic and magnetic forces the microscope imposes
on it. The wandering isn't a great distance mind you, just a few cm this way
or that, tending to fluctuate in a roughly diagonal path across the screen.
The drift is mostly a very smooth and slow motion, but occasionally the beam
will jump 5mm or so, and then continue on its lazy drift. Also, it doesn't
seem to drift back and forth from one point to another, just away from
center when you place it there with the beam shifts. It then goes off in one
direction and stays put when it moves as far as it wants/needs to.

This causes a bit of a problem when imaging at higher mags as you might
imagine. We suspect that we may have a box of questionable filaments, since
we are only getting between 60 and 140 hours out of them without any obvious
change in microscope operating conditions. (Gun vacuum seems good at around
4x10^-7 mbar.)
Are there any other obvious things things I can check that might be causing
this? Help me pin down my wayward beam!

Thanks,
--
Bradford Ross

Microscopy Technician
BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996


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From: baskin-at-bio.umass.edu
Date: Tue, 10 Aug 2010 17:56:19 -0500
Subject: [Microscopy] TEM sample storage before embed

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Greetings,
I am asking this question on behalf of a graduate student in
my department, Carrie Thurber.

Carrie needs to store a large number of rice florets for 0ne
half to one year prior to embedding in Epon Araldite. The eventual
purpose is to examine histology with standard heavy metal staining in
TEM. She is particularly interested in certain abscission layers.
Harvesting will be done in the greenhouse, attached to the lab (so
heroics for field sampling are not required). For her usual preps
(ie, those that do not require storage), she fixes in buffered glut,
rinses, dehydrates in ethanol and so on. Is it possible to put the
samples in a fixative (FAA, if not glut??) and leave them in the cold
for the duration? Or take them out of fix and rinse and store in an
aqueous buffer? If anyone has a good protocol for this, Carrie will
be delighted. Her e-mail is cthurber-at-cns.umass.edu. Please be sure to
cc her because she is not a member of this list.
Many thanks,
Tobias


--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Biology Department
/_ / __ /__ \ \ \__ 611 N. Pleasant St.
/ / / \ \ \ University of Massachusetts
/ / / \ \ \ Amherst, MA, 01003
/ / ___ / \ \__/ \ ____
www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

==============================Original Headers==============================
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From: sjrobin-at-illinois.edu
Date: Tue, 10 Aug 2010 19:27:34 -0500
Subject: [Microscopy] viaWWW: Microscopist/Spectroscopist Job Posting

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Email: sjrobin-at-illinois.edu
Name: Scott J. Robinson

Organization: Beckman Institute, University of Illinois

Title-Subject: [Filtered] Microscopist/Spectroscopist Job Posting

Message: Light Microscopist/Spectroscopist
The Imaging Technology Group at the Beckman
Institute for Advanced Science and Technology,
University of Illinois at UrbanañChampaign, is
searching for an energetic, knowledgeable, and
personable individual to fill the position of
Light Microscopist/Spectroscopist in the
Microscopy Suite.

The Light Microscopist/Spectroscopist provides
direct support (training, consultation, help with
optimization of imaging/spectroscopy) to
Microscopy Suite users (mostly graduate students
and postdoctoral researchers) in all disciplines
(primarily biology, materials, and biomaterials)
as well as maintenance, enhancement, and planning
for a wide range of optical imaging (laser
scanning confocal microscopy, multiphoton
microscopy, fluorescence microscopy, reflected
light microscopy, computer-based stereology and
nerve-tracing on an upright microscope) and
spectroscopy (UV/Vis/NIR and Raman, fluorescence
correlation spectroscopy [FCS]) instrumentation.

For full consideration, applications must be
received by August 31, 2010. To view the full job
posting and apply to this position, please visit
the Illinois Job Board at
https://jobs.illinois.edu/. UIUC is an EEAAO
Employer.

Thank you
Scott Robinson
--
Microscopy Suite | Imaging Technology Group
Beckman Institute for Advanced Science and Technology
University of Illinois at Urbana-Champaign
405 North Mathews Avenue, Urbana IL 61801
217 265 5071 217 244 6219 (fax)
sjrobin-at-illinois.edu http://itg.beckman.illinois.edu/


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From: andrewb-at-vsl.cua.edu
Date: Tue, 10 Aug 2010 20:55:55 -0500
Subject: [Microscopy] Mapping and "Micro"analysis on Rigaku Primus II XRF

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

Fellow Listserver Members,
We at Catholic University are in the final process of evaluating
wavelength XRF spectrometers in preparation for a purchase to replace our
present unit. Although the majority of our work is bulk samples, The Rigaku
Primus II has a mapping option available with a resolution down to 0.5 mm
which has some appeal to me, perhaps less for the mapping capability than
for the ability to do coarse resolution microanalysis on some of our glass
samples which have already been prepared for SEM work. As you all know,
mobile akali ions is glasses present a chalenge to analyze accurately under
the electron beam because of space charge induced migration. I'm
wondering if there any users of this unit on the list, and if they would be
willing to share their experience with the mapping/microanalysis feature?
Also, comments on ease of use, reliability, software adequacy, and Rigaku's
service support would be welcome as we don't have much of a user base to
contact in our immediate geographical area. Please reply directly to me off-
list. Thanks in advance.


Sincerely yours,
Andy Buechele
Andrew C. Buechele, Ph.D.
The Catholic University of America - VSL
409 Hannan Hall
Washington, D.C. 20064
Phone: 202-319-4995 Fax: 202-319-4469


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From: protrain-at-emcourses.com
Date: Wed, 11 Aug 2010 03:13:55 -0500
Subject: [Microscopy] TEM beam drift after filament change.

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

Hi

This looks like a classic charge situation. There must be a piece of dirt
charging in your column which will act as a deflection coil as the charge
grows and as it discharges the beam will fall back to its original position.
The more erratic the movement is the poorer the contact between the dirt and
the column; a distinction between a hair and a lump of material.

A column clean would be my suggestion.

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: bnross-at-interchange.ubc.ca [mailto:bnross-at-interchange.ubc.ca]
Sent: 10 August 2010 21:54
To: protrain-at-emcourses.com

Hello fellow listserver colleagues,

I installed a new filament in our Hitachi H7600 yesterday and all seemed
well after gun alignment etc. However, today it appears that the beam just
wants to wander its merry way around the screen as if we were merely
suggesting it stayed centered, rather than it being steadfastly forced into
place by the strict electrostatic and magnetic forces the microscope imposes
on it. The wandering isn't a great distance mind you, just a few cm this way
or that, tending to fluctuate in a roughly diagonal path across the screen.
The drift is mostly a very smooth and slow motion, but occasionally the beam
will jump 5mm or so, and then continue on its lazy drift. Also, it doesn't
seem to drift back and forth from one point to another, just away from
center when you place it there with the beam shifts. It then goes off in one
direction and stays put when it moves as far as it wants/needs to.

This causes a bit of a problem when imaging at higher mags as you might
imagine. We suspect that we may have a box of questionable filaments, since
we are only getting between 60 and 140 hours out of them without any obvious
change in microscope operating conditions. (Gun vacuum seems good at around
4x10^-7 mbar.)
Are there any other obvious things things I can check that might be causing
this? Help me pin down my wayward beam!

Thanks,
--
Bradford Ross

Microscopy Technician
BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996


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From: nizets2-at-yahoo.com
Date: Wed, 11 Aug 2010 08:19:45 -0500
Subject: [Microscopy] viaWWW: colocalization software

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

Hi Molly,

I hope I understood you well, if not please apologize.
If your protein is expected to associate but not enter the sphere, it is no
surprise that ImageJ cannot colocalize them, because perhaps they simply do not
colocalize! Depending on the size of the sphere and the scale of the
interaction, it may simply be that both signals are next to each other. On the
good side, demonstrating that 2 signals are consistently next to each other may
be enough to indicate that there is an interaction.
Sorry if I did not directly reply to your question.

Best regards,
Stephane

 


----- Original Message ----
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To: nizets2-at-yahoo.com
Sent: Fri, August 6, 2010 7:17:48 PM

This Question/Comment was submitted to the Microscopy Listserver
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Email: mshaw1-at-lumc.edu
Name: Molly Shaw

Organization: Loyola University Chicago

Title-Subject: [Filtered] colocalization software

Message: Hi there,

My lab is looking to purchase colocalization software, so I was
wondering if any of you have recommendations. We are using a Zeiss
LSM 510, so we don't need imaging capabilities, just colocalization
quantification.

We have been using ImageJ, but it doesn't seem to be able to do
everything we need. Our main issue is that we're trying to colocalize
a protein with a non-protein sphere. The protein does not actually
enter the sphere, but we know they do associate. However, this
association is not being reflected in the coefficients generated by
ImageJ (Pearson is only about 0.3). We're wondering if it's necessary
to use a program that could treat our protein as a sphere (or
something similar) and thus quantify colocalization more accurately.

Any suggestions are welcome.

Thank you,
Molly

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From: bnross-at-interchange.ubc.ca
Date: Wed, 11 Aug 2010 15:34:29 -0500
Subject: [Microscopy] Re: TEM beam drift after filament change.

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

Hello again,

Thanks to everyone for their suggestions! We had some people working late yesterday, so I didn't have a chance to open up the gun and check for dirt/fuzz/hair. However, after being left on standby overnight as it usually is, the scope (and beam) seem to be stable now. I know for sure that the anode and the column in general need a good cleaning, so I will get around to that after I'm back from vacation.

For summary's sake, the general consensus seemed to be something, (dirt/fuzz/hair etc.) in the gun or on the anode or one of the apertures that was charging and altering the path of the beam. Some people also agreed that it could be a bad filament with a poor solder joint/weld to the posts in the insulator. A loose set screw holding the filament in place was also mentioned as a possibility.

Thanks again,
--
Bradford Ross

Microscopy Technician
BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996


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From: ibarke2-at-uwo.ca
Date: Thu, 12 Aug 2010 11:08:29 -0500
Subject: [Microscopy] Sample Prep- EBSD Vibratory Polisher - Alumina Suspension Growing

Contents Retrieved from Microscopy Listserver Archives
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Hi there,
Recently, we polished some samples for EBSD analysis using a Buehler Vibratory Polisher and a solution of MasterPrep Agglomerate free Seeded-gel Alumina Suspension (0.05micron pH8.5).  I've been told that after we pour in the solution into the polishing bowl, we can just keep the microcloth pad wet and reuse the solution that's on the pad on a later date.  It has been a couple weeks since we last used the pad (however we have been keeping it wet) and now there's a black mold creeping across the surface of the polishing solution. Is this solution still usable, or should we dump it and replace the cloth?
Please reply to ibarke2-at-uwo.ca
Thank you.



Ivan R. Barker
UWO Earth Sciences MSc
ZAPLab Geochronology Research Technician
T: 519-661-2111 ext. 88397
E: ibarke2-at-uwo.ca
 





==============================Original Headers==============================
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7, 20 -- Subject: Sample Prep- EBSD Vibratory Polisher - Alumina Suspension Growing
7, 20 -- Mold?
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From: smalinskas-at-yahoo.com
Date: Thu, 12 Aug 2010 13:10:27 -0500
Subject: [Microscopy] Re: Sample Prep- EBSD Vibratory Polisher - Alumina Suspension Growing

Contents Retrieved from Microscopy Listserver Archives
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Ivan,

I've used a vibratory polisher for years and never had a problem with mold. All I used was levigated 0.05 micron) alumina and water. I let it dry out between each use, and replenished with water just before using. I don't believe you have to keep it wet.

Stu Smalinskas, P.E.
Metallurgist
SKF
(734) 414-6862

--- On Thu, 8/12/10, ibarke2-at-uwo.ca {ibarke2-at-uwo.ca} wrote:

} From: ibarke2-at-uwo.ca {ibarke2-at-uwo.ca}
} Subject: [Microscopy] Sample Prep- EBSD Vibratory Polisher - Alumina Suspension Growing
} To: smalinskas-at-yahoo.com
} Date: Thursday, August 12, 2010, 12:10 PM
}
}
}
} ----------------------------------------------------------------------------
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}
} Hi there,
} Recently, we polished some samples for EBSD analysis using
} a Buehler Vibratory Polisher and a solution of MasterPrep
} Agglomerate free Seeded-gel Alumina Suspension (0.05micron
} pH8.5).  I've been told that after we pour in the solution
} into the polishing bowl, we can just keep the microcloth pad
} wet and reuse the solution that's on the pad on a later
} date.  It has been a couple weeks since we last used the
} pad (however we have been keeping it wet) and now there's a
} black mold creeping across the surface of the polishing
} solution. Is this solution still usable, or should we dump
} it and replace the cloth?
} Please reply to ibarke2-at-uwo.ca
} Thank you.
}
}
}
} Ivan R. Barker
} UWO Earth Sciences MSc
} ZAPLab Geochronology Research Technician
} T: 519-661-2111 ext. 88397
} E: ibarke2-at-uwo.ca
}  






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9, 29 -- Subject: Re: [Microscopy] Sample Prep- EBSD Vibratory Polisher - Alumina Suspension Growing
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From: bbandli-at-d.umn.edu
Date: Thu, 12 Aug 2010 13:21:24 -0500
Subject: [Microscopy] Re: Sample Prep- EBSD Vibratory Polisher - Alumina Suspension Growing

Contents Retrieved from Microscopy Listserver Archives
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Hi Ivan,
I've been using a Buehler Vibratory polisher for EBSD prep with
colloidal silica (MasterMet 2) to polish mainly geological samples.
What I do is once I am finished with a batch of samples I use running
tap water to thoroughly rinse the bowl and cloth (I use adhesive backed
cloths and leave them in the bowl) and let dry. I store the bowl on the
machine with the cover over it until my next batch. Then I add fresh
polishing suspension and repeat the process. I've been able to use the
same cloth about a dozen times before it starts to show signs of wear
and/or diminished polishing quality.
Best,
Bryan

ibarke2-at-uwo.ca wrote:
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
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}
} Hi there,
} Recently, we polished some samples for EBSD analysis using a Buehler Vibratory Polisher and a solution of MasterPrep Agglomerate free Seeded-gel Alumina Suspension (0.05micron pH8.5). I've been told that after we pour in the solution into the polishing bowl, we can just keep the microcloth pad wet and reuse the solution that's on the pad on a later date. It has been a couple weeks since we last used the pad (however we have been keeping it wet) and now there's a black mold creeping across the surface of the polishing solution. Is this solution still usable, or should we dump it and replace the cloth?
} Please reply to ibarke2-at-uwo.ca
} Thank you.
}
}
}
} Ivan R. Barker
} UWO Earth Sciences MSc
} ZAPLab Geochronology Research Technician
} T: 519-661-2111 ext. 88397
} E: ibarke2-at-uwo.ca
}
}
}


--
~~~~~~~~~~~~~~~~~~~~
Bryan Bandli
SEM Laboratory Manager
University of Minnesota Duluth
Life Science 93

Mail:
UMD Geological Sciences
229 Heller Hall
1114 Kirby Dr.
Duluth, MN 55812-3036

Phone: 218-726-7362
Fax: 218-726-8275

www.d.umn.edu/SEM/


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From: Frank_Karl-at-lincolnelectric.com
Date: Thu, 12 Aug 2010 13:31:14 -0500
Subject: [Microscopy] Re: Sample Prep- EBSD Vibratory Polisher - Alumina Suspension

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

I would suggest picking a small droplet up and placing it on a glass slide
(and a coverslip!) and looking at it with the light microscope. If it's
mold, fungus, or other living stuff you should get some information. I'd
discard anything with mold or fungus...

I use to use this system for polishing ebonited rubber samples. When my
solutions dried out, the Al oxides formed little hard lumps that didn't
disperse. I'd dump the solution anytime it was going to sit for a day or
two.


stay safe..........
Frank Karl.........

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From: Adena.Rollins-at-sandisk.com
Date: Fri, 13 Aug 2010 08:35:16 -0500
Subject: [Microscopy] viaWWW: Ion-Slicer

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Email: Adena.Rollins-at-sandisk.com
Name: Adena Rollins

Organization: SanDisk

Title-Subject: [Filtered] Ion-Slicer

Message: Does anyone know of any service labs in the Silicon Valley
that have the JEOL ion-slicer or anything similar to that? We need
to cut a package of stacked die with a defect and the FIB technique
is taking too long!

Thank you for any information

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From: pmachado-at-igc.gulbenkian.pt
Date: Fri, 13 Aug 2010 08:35:47 -0500
Subject: [Microscopy] viaWWW: Ultramicrotome illumination system problem

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Email: pmachado-at-igc.gulbenkian.pt
Name: Pedro Machado

Organization: Instituto Gulbenkian de Ciencia

Title-Subject: [Filtered] Ultramicrotome illumination system problem

Message: Dear all,

Our ultramicrotome is a 14 years old Reichert Ultracut S (Leica).

It has some problems with the illumination system that the
manufacturers can't solve because it seems the needed parts are not
produced anymore.

I am trying to get from them which part of the illumination system is
really not working.

I was wondering if any of you had a similar problem and can give me
some advice.

I am also interested in buying a second hand similar model (as I have
the cryo setup for this ultramcrotome) if by chance you have one that
is not used anymore.

Thank you very much.

Kind regards,
Pedro Machado


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From: bozzola-at-siu.edu
Date: Fri, 13 Aug 2010 16:53:57 -0500
Subject: [Microscopy] SEM: current striking specimen

Contents Retrieved from Microscopy Listserver Archives
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Has anyone measured the current striking a specimen in the SEM? (We do
not have a meter to make such measurements.)

I realize this is rather vague and depends on kV, emission current,
spot size, vacuum and other variables. But, if someone has done such a
measurement and would provide the operational conditions, it would be
useful to know.

Thank you.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


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5, 18 -- Subject: SEM: current striking specimen
5, 18 -- From: John Bozzola {bozzola-at-siu.edu}
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From: bernard-at-berkeleyrc.com
Date: Fri, 13 Aug 2010 17:26:34 -0500
Subject: [Microscopy] Re: SEM: current striking specimen

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

} Has anyone measured the current striking a specimen in the SEM? (We do
} not have a meter to make such measurements.)

See Goldstein:
"Probe Current. The most straightforward quantity to measure is the
incident beam current i, since this can be done with a 'Faraday cup'
which is simply a container completely closed except for a small entrance
aperture (see Fig. 2.25)..."

You will, however, need an ammeter that measures less than a microamp,
commonly referred to as a picoammeter.

--
Bernard R. Cuzzillo, Ph.D., P.E.
Berkeley Research Company (BRC)
600 Addison Street
Berkeley, CA  94710-1920
USA

www.berkeleyrc.com

bernard-at-berkeleyrc.com

Cell phone: 510.821.2499
Office phone: 510.868.4333
Fax: 510.868.4351


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From: Jessica.Riesterer-at-empa.ch
Date: Mon, 16 Aug 2010 08:53:04 -0500
Subject: [Microscopy] help with stage drift in the FIB at tilt

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

Total current on the sample varies by several orders of magnitude, from less than a pA to a few μA.

Bearing in mind that these currents in some FE-SEMs can be going into very small spots, current density can be very high. Thinking about it, what's surprising is not that some samples damage but that so many survive!

On 13 Aug 2010, at 23:01, bozzola-at-siu.edu wrote:

}
}
}
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}
} Has anyone measured the current striking a specimen in the SEM? (We do
} not have a meter to make such measurements.)
}
} I realize this is rather vague and depends on kV, emission current,
} spot size, vacuum and other variables. But, if someone has done such a
} measurement and would provide the operational conditions, it would be
} useful to know.
}
} Thank you.
}
} --
} John J. Bozzola, Ph.D., Director
} I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
} Southern Illinois University
} 750 Communications Drive
} Carbondale, IL 62901
} Phone: 618-453-3730
}

Larry Stoter
(Working on a Microsoft-free computer)



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The primary electron beam current is dependent on the selections of
objective aperture, filament current (for thermal emission) or extractor
voltage (for field emission), condense and objective lens setting and beam
energy, etc.
The vacuum does not affect the primary electron beam current in a SEM. Spot
size only change the intensity (or brightness),not the beam current.

Dr. Xiaoping Zha
Department of Electronics
University of York

--------------------------------------------------
X-from: {bernard-at-berkeleyrc.com}
Sent: Friday, August 13, 2010 11:31 PM
To: {charlesping-at-hotmail.com}

I have a custom-built AFM that I mount inside our Tescan Lyra dual beam FIB. I mount it by removing the standard peg-type stage plate and screwing the mounting plate directly to the stage mechanism with two screws. Unfortunately, I have enough drift when tilted to 55 degrees that milling the same location is not possible after scanning with the AFM (30-40 minutes later). I know that I am well below the stage's weight limit at 0 degree tilt. Does anyone have suggestions on minimizing this drift?
Thanks!
Jessica

____________________________________________________
Jessica L. Riesterer, Ph.D.
Mechanics of Materials and Nanostructures Laboratory
Empa - Materials Science & Technology
Feuerwerkerstr. 39
CH-3602 Thun, Switzerland
Jessica.Riesterer-at-empa.ch
p:+41 33 228 46 07
f:+41 33 228 44 90
http://www.empa.ch/abt128
____________________________________________________





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From: benada-at-biomed.cas.cz
Date: Mon, 16 Aug 2010 08:58:11 -0500
Subject: [Microscopy] C100 trouble

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

Hello,
Our Philips CM100 does not start after late night blackout. I have checked all
fuses (F1 thru F10) on front panel of the mains cabinet and all were OK, but
all signal light and LEDs were off.
Auxiliary mains panel located down on left side of the mains cabinet is alive.
Please, can anybody give me some hints?

Thanking you in advance
Oldrich

--
Oldrich Benada
Institute of Microbiology
Acad. Sci. CR
Videnska 1024
CZ-142 20 Prague 4
Czech Republic

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From: kenconverse-at-qualityimages.biz
Date: Mon, 16 Aug 2010 09:26:04 -0500
Subject: [Microscopy] SEM: current striking specimen

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

Charles, I agree with the first part of your post, but I take exception with the spot size statement.

"Spot size" can change the beam current. It depends on the instrument. On my JEOL microprobe, spot size only changes the diameter of the beam. The current stays constant. On my new FEI SEM (and most other SEMs I know of), spot size does change beam current and my understanding is that it is synonymous with the condensor lens control. I rather wish in those cases the control was simply called beam current.

Does anyone know off the top of their head how closely "spot size" values as used on SEMs are related to the true spot size? Certainly, the values are without units and some ratio is needed, at least. Is the relation anything close to linear? I know that "beam current" on one of our SEMs is definitely NOT a linear relation to real beam current.

Warren Straszheim
Iowa State University
________________________________________
X-from: charlesping-at-hotmail.com [charlesping-at-hotmail.com]
Sent: Sunday, August 15, 2010 5:07 AM
To: wesaia-at-iastate.edu


The primary electron beam current is dependent on the selections of
objective aperture, filament current (for thermal emission) or extractor
voltage (for field emission), condense and objective lens setting and beam
energy, etc.
The vacuum does not affect the primary electron beam current in a SEM. Spot
size only change the intensity (or brightness),not the beam current.

Dr. Xiaoping Zha
Department of Electronics
University of York

--------------------------------------------------
X-from: {bernard-at-berkeleyrc.com}
Sent: Friday, August 13, 2010 11:31 PM
To: {charlesping-at-hotmail.com}

Hi John,
I would suggest making a Faraday cup and then beg, borrow or steal a
picoameter from Physics or Electrical Engineering for a day or so and make
yourself a table of various working conditions and the resulting current.
Don't forget that lens hysteresis must be normalized for reproducibility.

You haven't said why you need this info. If it's going to be important in
on-going work, buy a picoameter and permanently hook it up to your stage.
There are so many variables that frequent comparison readings are important.

As for using someone else's settings, you haven't said what instrument
you're using and settings from a different model from the same manufacturer
are going to give different results, let alone settings from a different
manufacturer. I would expect that you'll even get different results (but
closer) from the same model. There were some Kiethly meters on Ebay for
about $250.

Good luck!

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: bozzola-at-siu.edu [mailto:bozzola-at-siu.edu]
Sent: Friday, August 13, 2010 5:57 PM
To: kenconverse-at-qualityimages.biz

Has anyone measured the current striking a specimen in the SEM? (We do
not have a meter to make such measurements.)

I realize this is rather vague and depends on kV, emission current,
spot size, vacuum and other variables. But, if someone has done such a
measurement and would provide the operational conditions, it would be
useful to know.

Thank you.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


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From: bbandli-at-d.umn.edu
Date: Mon, 16 Aug 2010 10:06:19 -0500
Subject: [Microscopy] SEM: current striking specimen

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

Hello John,

As others, including yourself, have stated, the answer is "it depends"
and the range is from picoamps to microamps. But...

I have run our JEOL JSM-6490LV (W-filament) at a range of operating
conditions and measured the probe current with a stage mounted Faraday
cup and Kiethly picoammeter. The range is from ~1 pA -at- 5 kV (lowest
acc. volt I tested, 30 µm obj. aperture, and at the minimum condenser
lens setting, which isn't useful for imaging) to 3.5 µA -at- 30 kV (highest
acc. volt, 100 µm obj and max. condenser setting, which also isn't very
useful for imaging). Also, the current increases exponentially with
increasing spot size setting, not linearly.

Hope this helps,

Bryan

--
~~~~~~~~~~~~~~~~~~~~
Bryan Bandli
SEM Laboratory Manager
University of Minnesota Duluth
Life Science 93

Mail:
UMD Geological Sciences
229 Heller Hall
1114 Kirby Dr.
Duluth, MN 55812-3036

Phone: 218-726-7362
Fax: 218-726-8275

www.d.umn.edu/SEM/


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From: eschumacher-at-mccrone.com
Date: Mon, 16 Aug 2010 10:43:46 -0500
Subject: [Microscopy] Meeting Announcement: Midwest Microscopy and Microanalysis Society

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

Greetings All,

The Midwest Microscopy and Microanalysis Society will hold its fourth annual Optical Microscopy Workshop on Wednesday, September 29th, at the Hooke College of Applied Sciences (formerly the College of Microscopy) in Westmont, IL. A two-topic program will highlight optical microscopy of sands and minerals, and optical microscope QA/calibration. The day-long workshop will include presentations, demonstrations and hands-on instruction. Please check the Meetings page on our website for further details and registration information:

www.midwestmicroscopy.org

We look forward to seeing you on September 29th.

Sincerely,

Elaine Schumacher
Program Coordinator, M3S



*********************************************************************
Elaine F.Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com



*********************************************************************
This message and any attachments are solely for the
intended recipient. If you are not the intended recipient,
disclosure, copying, use or distribution of the information
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From: vitalylazar-at-att.net
Date: Mon, 16 Aug 2010 12:06:13 -0500
Subject: [Microscopy] Re: C100 trouble

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

Open the door on the left side of power supply cabinet and see a number
of LED lights - all should be off until you depress Mains ON button on
TEM control panel.

Watch LED lights carefully while pressing that button. The (all) 4 green
lights on the top row must illuminate. These are +5V; +24V; +15V; and
-15V power supplies. Additionally 2 green LEDs must illuminate on the
next row down (+/- 30V doubler).

Any of the upper 4 lights remaining off or illuminating momentarily and
then going off indicate(s) defective power supply that need(s) replacement.

The above is the most likely problem but is not the only one possible.

Vitaly Feingold
SIA
2773 Heath Lane
Duluth GA 30096
Ph. 770-232-7785
Fax 770-232-1791
www.sia-cam.com
vitaly-at-sia-cam.com

On 8/16/2010 9:59 AM, benada-at-biomed.cas.cz wrote:
} ----------------------------------------------------------------------------
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} Hello,
} Our Philips CM100 does not start after late night blackout. I have checked all
} fuses (F1 thru F10) on front panel of the mains cabinet and all were OK, but
} all signal light and LEDs were off.
} Auxiliary mains panel located down on left side of the mains cabinet is alive.
} Please, can anybody give me some hints?
}
} Thanking you in advance
} Oldrich
}

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From: fahayes-at-ucdavis.edu
Date: Mon, 16 Aug 2010 15:14:59 -0500
Subject: [Microscopy] how to properly cleave a Si wafer

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

Whats the best way to cleave a Si wafer for edge retention on a fractured
edge?

Fred Hayes
UC Davis


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From: s.h.coetzee-at-gmail.com
Date: Tue, 17 Aug 2010 05:28:09 -0500
Subject: [Microscopy] SEM: current striking specimen

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

Hi

Spot size and probe current values specified on the microscope display in
SEM are often a joke!

Changing the filament position, the bias setting and the accelerating
voltage all change the size of the virtual source. This source is the
starting point for the condenser system, which simply demagnifies the source
up to about 10,000X. However change the source size and the condenser
system has no indication of the change and therefore will be unable to
compensate.

The instrument designer starts at the electron gun and positions the
filament in what he believes is the perfect position; how many operators
know which is the perfect position? At this setting the spot size and probe
current readouts are as near to correct as they will ever be. Only if the
manufacturer programs their control system to compensate for an accelerating
voltage change under these conditions, will any fixed figure of spot size or
probe current remain correct.

Designers run the instrument much harder than the average operator in the
vast majority of cases. Running the instrument harder means a different
filament position and a different bias setting, both of which change the
virtual source size, the spot size and of course the probe current. The
designer is after resolution and convenience but is unable to predict how
the day to day operator will use the instrument.

Should you want to calibrate the spot size for future use you will first
need to set a standard filament operating position and decide upon an
emission current and therefore a bias setting. However you should calibrate
at each of the accelerating voltages that will be used.

Now with a FEG when you change accelerating voltage the source size also
tends to change, once again making spot size and probe current incorrect
unless the manufacturer has adjusted the condenser system to compensate, but
perhaps that is asking for too much?

When I teach students, to deflect them away from believing the data
presented by the manufactures, I tell them to consider the readout is in
bananas. You use the same number of bananas to obtain the same quality of
result, under the same conditions, on the same sample in the future, that is
the best use of the information provided on spot size or probe current.

The only way to trust a probe current reading is to use a Faraday Cup!

I am happy for a manufacturer to shoot me down if I am leading you all
astray, because my days of involvement in design are over?

Regards

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com





-----Original Message-----
X-from: wesaia-at-iastate.edu [mailto:wesaia-at-iastate.edu]
Sent: 16 August 2010 15:03
To: protrain-at-emcourses.com

Charles, I agree with the first part of your post, but I take exception with
the spot size statement.

"Spot size" can change the beam current. It depends on the instrument. On my
JEOL microprobe, spot size only changes the diameter of the beam. The
current stays constant. On my new FEI SEM (and most other SEMs I know of),
spot size does change beam current and my understanding is that it is
synonymous with the condensor lens control. I rather wish in those cases the
control was simply called beam current.

Does anyone know off the top of their head how closely "spot size" values as
used on SEMs are related to the true spot size? Certainly, the values are
without units and some ratio is needed, at least. Is the relation anything
close to linear? I know that "beam current" on one of our SEMs is definitely
NOT a linear relation to real beam current.

Warren Straszheim
Iowa State University
________________________________________
X-from: charlesping-at-hotmail.com [charlesping-at-hotmail.com]
Sent: Sunday, August 15, 2010 5:07 AM
To: wesaia-at-iastate.edu


The primary electron beam current is dependent on the selections of
objective aperture, filament current (for thermal emission) or extractor
voltage (for field emission), condense and objective lens setting and beam
energy, etc.
The vacuum does not affect the primary electron beam current in a SEM. Spot
size only change the intensity (or brightness),not the beam current.

Dr. Xiaoping Zha
Department of Electronics
University of York

--------------------------------------------------
X-from: {bernard-at-berkeleyrc.com}
Sent: Friday, August 13, 2010 11:31 PM
To: {charlesping-at-hotmail.com}

I cannot agree more with Steve.
You can make a "cheap" faraday cup by drilling a 1mm hole in a stub
without going through the stub and glue a used Pt aperture with a hole
diameter of 500 micron or less over the hole. Relatively Cheap pico
amp meters are available on the market. Then there is another matter.
Some days the beam stabilize in ~30 min (less than 5% drift per
15min) and some days under the same settings with the same operator,
same holder, same sample, same filament same acceleration voltage it
can take up to 90 min to get the same stability.
That is microscopy;)

Stephan H Coetzee
EM Scientist
Electron microscope Unit
University of Botswana
Gaborone
Tell: +267 355 5222/2462
coetzees-at-mopipi.ub.bw


On Mon, Aug 16, 2010 at 10:27 PM, {protrain-at-emcourses.com} wrote:
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi
}
} Spot size and probe current values specified on the microscope display in
} SEM are often a joke!
}
} Changing the filament position, the bias setting and the accelerating
} voltage all change the size of the virtual source.  This source is the
} starting point for the condenser system, which simply demagnifies the source
} up to about 10,000X.  However change the source size and the condenser
} system has no indication of the change and therefore will be unable to
} compensate.
}
} The instrument designer starts at the electron gun and positions the
} filament in what he believes is the perfect position; how many operators
} know which is the perfect position? At this setting the spot size and probe
} current readouts are as near to correct as they will ever be.  Only if the
} manufacturer programs their control system to compensate for an accelerating
} voltage change under these conditions, will any fixed figure of spot size or
} probe current remain correct.
}
} Designers run the instrument much harder than the average operator in the
} vast majority of cases.  Running the instrument harder means a different
} filament position and a different bias setting, both of which change the
} virtual source size, the spot size and of course the probe current. The
} designer is after resolution and convenience but is unable to predict how
} the day to day operator will use the instrument.
}
} Should you want to calibrate the spot size for future use you will first
} need to set a standard filament operating position and decide upon an
} emission current and therefore a bias setting. However you should calibrate
} at each of the accelerating voltages that will be used.
}
} Now with a FEG when you change accelerating voltage the source size also
} tends to change, once again making spot size and probe current incorrect
} unless the manufacturer has adjusted the condenser system to compensate, but
} perhaps that is asking for too much?
}
} When I teach students, to deflect them away from believing the data
} presented by the manufactures, I tell them to consider the readout is in
} bananas.  You use the same number of bananas to obtain the same quality of
} result, under the same conditions, on the same sample in the future, that is
} the best use of the information provided on spot size or probe current.
}
} The only way to trust a probe current reading is to use a Faraday Cup!
}
} I am happy for a manufacturer to shoot me down if I am leading you all
} astray, because my days of involvement in design are over?
}
} Regards
}
} Steve
}
} Steve Chapman FRMS
} Senior Consultant Protrain
} For consultancy and training in electron microscopy world wide
} Tel +44 (0)1280 816512  Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
} www.emcourses.com
}
}
}
}
}
} -----Original Message-----
} X-from: wesaia-at-iastate.edu [mailto:wesaia-at-iastate.edu]
} Sent: 16 August 2010 15:03
} To: protrain-at-emcourses.com
} Subject: [Microscopy] SEM: current striking specimen
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Charles, I agree with the first part of your post, but I take exception with
} the spot size statement.
}
} "Spot size" can change the beam current. It depends on the instrument. On my
} JEOL microprobe, spot size only changes the diameter of the beam. The
} current stays constant. On my new FEI SEM (and most other SEMs I know of),
} spot size does change beam current and my understanding is that it is
} synonymous with the condensor lens control. I rather wish in those cases the
} control was simply called beam current.
}
} Does anyone know off the top of their head how closely "spot size" values as
} used on SEMs are related to the true spot size? Certainly, the values are
} without units and some ratio is needed, at least. Is the relation anything
} close to linear? I know that "beam current" on one of our SEMs is definitely
} NOT a linear relation to real beam current.
}
} Warren Straszheim
} Iowa State University
} ________________________________________
} X-from: charlesping-at-hotmail.com [charlesping-at-hotmail.com]
} Sent: Sunday, August 15, 2010 5:07 AM
} To: wesaia-at-iastate.edu
} Subject: [Microscopy] Re: SEM: current striking specimen
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
}
} The primary electron beam current is dependent on the selections of
} objective aperture, filament current (for thermal emission) or extractor
} voltage (for field emission), condense and objective lens setting and beam
} energy, etc.
} The vacuum does not affect the primary electron beam current in a SEM. Spot
} size only change the intensity (or brightness),not the beam current.
}
} Dr. Xiaoping Zha
} Department of Electronics
} University of York
}
} --------------------------------------------------
} X-from: {bernard-at-berkeleyrc.com}
} Sent: Friday, August 13, 2010 11:31 PM
} To: {charlesping-at-hotmail.com}
} Subject: [Microscopy] Re: SEM: current striking specimen
}
} }
} }
} }
} }
} ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} } To  Subscribe/Unsubscribe --
} } http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} }
} ----------------------------------------------------------------------------
} }
} } } Has anyone measured the current striking a specimen in the SEM? (We do
} } } not have a meter to make such measurements.)
} }
} } See Goldstein:
} } "Probe Current. The most straightforward quantity to measure is the
} } incident beam current i, since this can be done with a 'Faraday cup'
} } which is simply a container completely closed except for a small entrance
} } aperture (see Fig. 2.25)..."
} }
} } You will, however, need an ammeter that measures less than a microamp,
} } commonly referred to as a picoammeter.
} }
} } --
} } Bernard R. Cuzzillo, Ph.D., P.E.
} } Berkeley Research Company (BRC)
} } 600 Addison Street
} } Berkeley, CA?94710-1920
} } USA
} }
} } www.berkeleyrc.com
} }
} } bernard-at-berkeleyrc.com
} }
} } Cell phone: 510.821.2499
} } Office phone: 510.868.4333
} } Fax: 510.868.4351
} }
}
}
} ==============================Original Headers==============================
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From: dufresne-at-ms.umanitoba.ca
Date: Tue, 17 Aug 2010 08:16:42 -0500
Subject: [Microscopy] viaWWW: LKB Pyramitome 11800

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Email: dufresne-at-ms.umanitoba.ca
Name: Andre Dufresne

Organization: University of Manitoba

Title-Subject: [Filtered] Pyramitome

Message: Hi All.
I was wondering if anyone might the instruction manual for a LKB
Pyramitome 11800. If so, E-mail me and we can make arragements to
get a copy. Cheers.

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From: delannoy-at-jhmi.edu
Date: Tue, 17 Aug 2010 08:23:48 -0500
Subject: [Microscopy] IEM bacteria

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Hello Microscopists,
I have a client that wants to do the following:
use anti-cAMP antibodies and electron
microscopy to visualize where cAMP localizes in the bacterium.
Question is will an LR white (post-embed labeling) work with this?
Any comments appreciated.

M Delannoy

==============================Original Headers==============================
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From: Cross-at-tru.ca
Date: Tue, 17 Aug 2010 12:46:34 -0500
Subject: [Microscopy] Re: IEM bacteria

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Hello! I have definitely had LR White work for both plant material and gram-positive bacteria. I used LR white Medium grade, accelerator hardened. Standard prep. Don't have the bacteria paper published yet, but here is a link to our plant one (http://onlinelibrary.wiley.com/doi/10.1111/j.1438-8677.2009.00223.x/pdf ); same general idea. Worked like a charm both times.

Cheers,
Cindy

} } } {delannoy-at-jhmi.edu} 08/17/10 6:34 AM } } }



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Hello Microscopists,
I have a client that wants to do the following:
use anti-cAMP antibodies and electron
microscopy to visualize where cAMP localizes in the bacterium.
Question is will an LR white (post-embed labeling) work with this?
Any comments appreciated.

M Delannoy

==============================Original Headers==============================
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2, 29 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
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==============================Original Headers==============================
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From: kret-at-ifpan.edu.pl
Date: Wed, 18 Aug 2010 07:05:16 -0500
Subject: [Microscopy] viaWWW: Postdoctoral position

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Email: kret-at-ifpan.edu.pl
Name: Slawomir Kret

Organization: Institute of Physics Polish Academy of Science in Warsaw, Poland

Title-Subject: [Filtered] Postdoctoral position
ìAdvanced TEM study of local electronic
properties of indium rich nitrides.î

Message: Postdoctoral position in Laboratory of
X-Ray and Electron Microscopy Research in
Institute of Physics Polish Academy of Science in
Warsaw, Poland,
ìAdvanced TEM study of local electronic properties of indium rich nitrides.î
More informations:
http://info.ifpan.edu.pl/index_en.php


**************************************************
dr Sławomir Kret
Instytut Fizyki PAN
Al. LotnikÛw 32/46
02-668 Warszawa
Poland
(+48-22)-8436601-3382
http://tem.ifpan.edu.pl
http://info.ifpan.edu.pl/index_en.php
Email: kret-at-ifpan.edu.pl
**************************************************



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From: gunter.moeller-at-arkemagroup.com
Date: Wed, 18 Aug 2010 07:05:46 -0500
Subject: [Microscopy] viaWWW: Safe handling of LN2 for EDS

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Email: gunter.moeller-at-arkemagroup.com
Name: Gunter Moeller

Organization: Arkema Inc.

Title-Subject: [Filtered] Safe handling of LN2 for EDS

Message: Hello,

I'm in the process of writing a safe operating procedure (SOP) for
filling EDS dewars with liquid nitrogen.

How do you minimize safety risks associated with LN2 in your lab? Do
you know of any information resources regarding safe handling of LN2
for EDS systems?

If there's any information you would like to share I'd love to hear from you.

Thanks,

Gunter Moeller

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From: TindallR-at-missouri.edu
Date: Wed, 18 Aug 2010 13:39:55 -0500
Subject: [Microscopy] TEM: Dilated Intercellular Space Travel

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Dear Collective,

One of our esteemed clients is wondering if dilated intercellular spaces in his tissue samples (horse hoof) might be due to chemical fixation artifact or if it's the real thing. I seem to recall hearing something in a presentation once about increased spacing between cells or separation of membranes showing up in conventional samples, but not in high-pressure frozen and freeze-substituted ones of the same tissue. Due to increased dilation of my mental membranes over time, I'm no longer sure....

A quick Google search indicates that most researchers seem to think that the dilation is real, rather than artifact, and might be diagnostic of certain conditions in esophageal tissue. I'm not aware of much (or anything) out there on hoof, but I'll keep looking.

In the meantime, any thoughts are welcome. Many thanks, as usual!

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




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From: wtivol-at-verizon.net
Date: Wed, 18 Aug 2010 15:19:21 -0500
Subject: [Microscopy] Re: viaWWW: Safe handling of LN2 for EDS

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On Aug 18, 2010, at 5:21 AM, gunter.moeller-at-arkemagroup.com wrote:

} This Question/Comment was submitted to the Microscopy Listserver
} using the WWW based Form at
} http://microscopy.com/MicroscopyListserver/MLFormMail.html
} ---------------------------------------------------------------------------
} Remember this posting is most likely not from a Subscriber, so when
} replying
} please copy both gunter.moeller-at-arkemagroup.com as well as the
} MIcroscopy Listserver
} ---------------------------------------------------------------------------
}
} Email: gunter.moeller-at-arkemagroup.com
} Name: Gunter Moeller
}
} Organization: Arkema Inc.
}
} Title-Subject: [Filtered] Safe handling of LN2 for EDS
}
} Message: Hello,
}
} I'm in the process of writing a safe operating procedure (SOP) for
} filling EDS dewars with liquid nitrogen.
}
} How do you minimize safety risks associated with LN2 in your lab? Do
} you know of any information resources regarding safe handling of LN2
} for EDS systems?
}
} If there's any information you would like to share I'd love to hear
} from you.
}
} Thanks,
}
} Gunter Moeller


Dear Gunter,
There are two ways that LN2 can potentially cause harm, cooling of
skin, and displacement of oxygen. The latter is unlikely to be an
issue for filling EDS dewars from small LN2 containers (4 liters,
say), but is a concern when filling the small LN2 container from a
large supply container (} 100 liters). Make sure that there is plenty
of ventilation so that the LN2 vapor can be diluted sufficiently into
the ambient air space, and be sure that vapor near the ground is well
mixed, since if one collapses due to lack of oxygen, a pool of cold N2
vapor on the floor can be fatal. In many places I've worked fire
regulations required that large LN2 containers be placed inside small
rooms, which is dangerous unless the door can be left open to a much
larger space, such as a hallway. The danger from LN2 cooling the skin
is not severe unless the liquid is held against the skin for an
extended time. This can be minimized by wearing clothing that has no
cuffs or other features that can trap the LN2. Sandals that are form-
fitted to the foot have depressions in which LN2 can collect, for
example. It is important to train everyone who will be filling the
EDS system how to recognize when N2 vapor can collect and when LN2 can
be trapped. Common sense should do the rest.
Yours,
Bill


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From: raristau-at-ims.uconn.edu
Date: Wed, 18 Aug 2010 16:58:50 -0500
Subject: [Microscopy] Re: viaWWW: Safe handling of LN2 for EDS

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

Gunter

I presume that you already are familiar with safe handling of LN2, but the
need to write the SOP has you concerned about getting the details just
right.

Entering the terms "safe handling ln2" in a popular search engine brought up
several promising links to government, and commercial suppliers of cryogenic
liquids safety guides. You may do well to have a look at some of these.

What might be missing in some of these guides are the dumb things that
people do in actual practice. A couple of notable ones that I can think of:

-Storing LN2 in a tightly closed container will result in the container
bursting. The actual force of the explosion is directly related to the
strength of the container.

-I have received painful burns from small drops of LN2 splashing onto hands
and arms although it was in contact with skin for very brief time. Much more
dangerous is getting clothing wet with LN2: the cold liquid is then in
prolonged contact with the skin. Proper cryogenic gloves, aprons and
footwear are always recommended.

-Some have the practice of demonstrating the "fun" effects of LN2 to novice
or young audiences. For example, freezing and eating marshmellows; pouring
LN2 on the hands; I even witnessed one fellow "drink" LN2. These practices
can give a very misleading message about LN2 safety.

I prefer to treat LN2 as any other potentially dangerous substance: Proper
safety clothing (e.g. close-fitting goggles--NOT safety glasses), proper
handling technique at all times, and proper application of its uses. (OK, I
will indulge in a demonstration of dipping a flower in LN2 and then breaking
it like glass. But this can also give a strong message about the dangers of
mishandling: "Just imagine your hand breaking like glass!")

Cheers and Safe Handling,
Roger



==============================Original Headers==============================
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From: arnec-at-bio.umass.edu
Date: Wed, 18 Aug 2010 17:38:45 -0500
Subject: [Microscopy] viaWWW: Nikon Diaphot slider for multiple cubes

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Email: arnec-at-bio.umass.edu
Name: Arne Christensen

Title-Subject: [Filtered] Nikon Diaphot slider for multiple cubes

Message: Dear listeners,

We have a Nikon Diaphot equipped with a slider that can hold one
filter set at a time. To facilitate switching filter sets, we would
like to purchase a slider that can hold multiple cubes.

Can anyone recommend a good source?

Thanks in advance,
A

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From: nizets2-at-yahoo.com
Date: Thu, 19 Aug 2010 01:56:08 -0500
Subject: [Microscopy] Re: viaWWW: Safe handling of LN2 for EDS

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

I've never ever been burnt by splashes of LN2 on my skin. Actually I prefer to
have LN2 directly on my skin than got caught inside clothes (or "protections").
I am talking of indirect splashes here, not directly pouring LN2 on the skin,
which may be very different and also requires much less care and attention. One
principle to keep in mind in that considering the huge difference of temperature
between the body and LN2, the liquid will evaporate instantly (as long as the
volume is limited), so the vapor needs a way out.
We also have lots of SOPs here, it seems to me that the only SOP missing is a
SOP for opening doors! ;-)

In summary, here are important things to consider:

- Educate the personal about the manipulation and potential danger of LN2. Only
educated personal should be allowed to manipulate LN2.
- LN2 containers should be stored in a ventilated area (cold chambers are not a
good idea).
- Always wears glasses, but those one with the sides open. Always wear cryogenic
gloves. Don't use closed shoes, I would recommend the ones with a holy upper
part (I had to struggle to get these shoes because the chemists want closed
shoes, which make sense when one works with chemicals or radioactivity)
- Don't pour at head height. I am 2 meters tall which brings my visage exactly
at the height of the EDS container. So I climb on a chair to fill in so I am
higher than the liquid (which tends to fall down as you probably know).
- Don't climb on unstable furniture, like chairs, to fill in the container ;-)
- Use common sense and keep concentrated. This includes : thinking before acting
(don't stop while acting though!) and don't do stupid things like :
    * think about the last evening you spent with your girlfriend while pouring
    * try to get the last drop by shaking the container violently
    * verify the height of the liquid with your finger (the finger of your
colleague is not a good idea, either) or any other body part
    * pour LN2 on hands to see what it does
    * look somewhere else while pouring LN2

Basically (don't write this in your SOP) one could really handle LN2 naked if
correctly educated and concentrated. The protections are there to limit the
consequences of human mistakes.

Happy SOPing

Stephane

 


----- Original Message ----
X-from: "raristau-at-ims.uconn.edu" {raristau-at-ims.uconn.edu}
To: nizets2-at-yahoo.com
Sent: Thu, August 19, 2010 12:02:55 AM

Gunter

I presume that you already are familiar with safe handling of LN2, but the
need to write the SOP has you concerned about getting the details just
right.

Entering the terms "safe handling ln2" in a popular search engine brought up
several promising links to government, and commercial suppliers of cryogenic
liquids safety guides. You may do well to have a look at some of these.

What might be missing in some of these guides are the dumb things that
people do in actual practice. A couple of notable ones that I can think of:

-Storing LN2 in a tightly closed container will result in the container
bursting. The actual force of the explosion is directly related to the
strength of the container.

-I have received painful burns from small drops of LN2 splashing onto hands
and arms although it was in contact with skin for very brief time. Much more
dangerous is getting clothing wet with LN2: the cold liquid is then in
prolonged contact with the skin. Proper cryogenic gloves, aprons and
footwear are always recommended.

-Some have the practice of demonstrating the "fun" effects of LN2 to novice
or young audiences. For example, freezing and eating marshmellows; pouring
LN2 on the hands; I even witnessed one fellow "drink" LN2. These practices
can give a very misleading message about LN2 safety.

I prefer to treat LN2 as any other potentially dangerous substance: Proper
safety clothing (e.g. close-fitting goggles--NOT safety glasses), proper
handling technique at all times, and proper application of its uses. (OK, I
will indulge in a demonstration of dipping a flower in LN2 and then breaking
it like glass. But this can also give a strong message about the dangers of
mishandling: "Just imagine your hand breaking like glass!")

Cheers and Safe Handling,
Roger



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From: jrminter-at-rochester.rr.com
Date: Thu, 19 Aug 2010 08:40:50 -0500
Subject: [Microscopy] Re: viaWWW: Safe handling of LN2 for EDS

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

I agree with Stephane here. Because human skin is significantly warmer than the boiling point of liquid nitrogen and the liquid is non-viscous, when a small amount of liquid nitrogen impinges on skin, one observes the Leidenfrost phenomenon - where an insulating gas layer is formed between the liquid drops and the skin. The important point is to make sure that the liquid nitrogen is not trapped near the skin but can evaporate. Most of the cryo gloves I have seen trap the liquid near the skin. When we fill Dewars, we use a "Hot hand" device - a rubber device with nubby standoffs to grab the cold hose. It has lots of open area where the fingers insert and if any liquid splashes, it can get out. We also wear safety glasses without side shields. I have been doing cryo-TEM since 1990 and filling EDS dewars since 1986, and have never been burned by liquid nitrogen but have had small splashes several times. In all cases I was following essentially the suggestions made by Stephane.

My understanding is that cases of where people have been burned with liquid nitrogen have all been where it was trapped near the skin. When I took Prof. Sitte's cryomicrotomy training in Austria, they told us of a case where two researchers were carrying a large Dewar of the liquid. One was wearing high boots with his pants tucked into the boot. The liquid sloshed and flowed into the boot and soaked his pants. The burn was very nasty. The course organizers admonished us to follow procedures quite similar to those recommended by Stephane.

I should add that liquid ethane is another matter entirely, Liquid ethane forms a more viscous liquid and has a much higher boiling point. Several years ago, I was attempting to equilibrate a gel on a grid in a humidity chamber for an hour before plunging into liquid ethane. Just after I plunged the grid into the liquid ethane, some overflowed from the reservoir into the surrounding liquid nitrogen, solidified and blocked the grid holder. I took the cryo-tweezers to grab the solidified ethane and toss it into my nearby exhaust line like I had done many times before. This time I made the mistake of crossing over my hand holding the tweezers with the grid and a small amount fell on my hand. I got a nasty burn. I never made that mistake again. By the way, we typically wear full face shields when preparing grids for cryo-TEM. This lets us breathe freely without sending humid air into the cryogen holder and because our cryogen holders are made from large open Dewars and we had one implode.

Best Regards,
John


On Aug 19, 2010, at 2:56 AM, nizets2-at-yahoo.com wrote:

}
}
}
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}
} I've never ever been burnt by splashes of LN2 on my skin. Actually I prefer to
} have LN2 directly on my skin than got caught inside clothes (or "protections").
} I am talking of indirect splashes here, not directly pouring LN2 on the skin,
} which may be very different and also requires much less care and attention. One
} principle to keep in mind in that considering the huge difference of temperature
} between the body and LN2, the liquid will evaporate instantly (as long as the
} volume is limited), so the vapor needs a way out.
} We also have lots of SOPs here, it seems to me that the only SOP missing is a
} SOP for opening doors! ;-)
}
} In summary, here are important things to consider:
}
} - Educate the personal about the manipulation and potential danger of LN2. Only
} educated personal should be allowed to manipulate LN2.
} - LN2 containers should be stored in a ventilated area (cold chambers are not a
} good idea).
} - Always wears glasses, but those one with the sides open. Always wear cryogenic
} gloves. Don't use closed shoes, I would recommend the ones with a holy upper
} part (I had to struggle to get these shoes because the chemists want closed
} shoes, which make sense when one works with chemicals or radioactivity)
} - Don't pour at head height. I am 2 meters tall which brings my visage exactly
} at the height of the EDS container. So I climb on a chair to fill in so I am
} higher than the liquid (which tends to fall down as you probably know).
} - Don't climb on unstable furniture, like chairs, to fill in the container ;-)
} - Use common sense and keep concentrated. This includes : thinking before acting
} (don't stop while acting though!) and don't do stupid things like :
} * think about the last evening you spent with your girlfriend while pouring
} * try to get the last drop by shaking the container violently
} * verify the height of the liquid with your finger (the finger of your
} colleague is not a good idea, either) or any other body part
} * pour LN2 on hands to see what it does
} * look somewhere else while pouring LN2
}
} Basically (don't write this in your SOP) one could really handle LN2 naked if
} correctly educated and concentrated. The protections are there to limit the
} consequences of human mistakes.
}
} Happy SOPing
}
} Stephane


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From: Marta.Jarquin-Pardo-at-STJUDE.ORG
Date: Thu, 19 Aug 2010 11:38:22 -0500
Subject: [Microscopy] Job Opening Cell Imaging Shred Resources St. Jude Children's

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

Hello,

Currently we have an opening for a Director of Light Microscopy (Job Number 18238) in the Cell and Tissue Imaging Shared Resource at St. Jude Children's Research Hospital

The Cell and Tissue Imaging (CTI) Shared Resource at St. Jude Children's Research Hospital combines technology with teamwork to facilitate studies that advance our understanding of fundamental biological processes and our ability to treat or prevent catastrophic diseases of childhood.

The Director of the Light Microscopy Facility is responsible for managing the light microscopy facility within the institution's Cell and Tissue Imaging (CTI) shared resource and for collaborating with faculty and staff on research projects. Light microscopy includes co focal laser scanning microscopy, multiphoton microscopy, live cell imaging and microinjection. The Director of the Light Microscopy Facility coordinates experimental design for advanced light microscopy; trains staff on experimental design, light microscopy and data processing and analysis; and maintains image archives.

Requirements:

A bachelor's degree in a relevant scientific field area and 10 years of laboratory experience utilizing advanced fluorescent technologies and instrumentation is required.

Demonstrated excellence in the management of a light microscopy resource facility is required. Demonstrated progression in continuing education in advanced light microscopy techniques is required.

TO FIND OUT MORE ABOUT THIS POSTION AN TO APPLY GO TO www.stjude.org/jobs

St. Jude offers a positive working culture, professional advancement and competitive compensation.

St. Jude is an Equal Opportunity Employer and a Drug-Free Workplace.

Candidates receiving offers of employment will be subject to pre-employment drug testing and background checks.

Regards,

Marta


Email Disclaimer: www.stjude.org/emaildisclaimer



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From: mcauliff-at-umdnj.edu
Date: Thu, 19 Aug 2010 12:45:19 -0500
Subject: [Microscopy] section adhesives for TEM grids

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

Having been visiting a dermatologist for the last 10 years or so to have pre-cancerous lesions removed by burning with liquid nitrogen I can tell you that the burn is tolerable if the LN2 is directed at a single spot for only one to two seconds but that is long enough for a blister to form. Beyond that, say about four seconds, the burn brings tears to one's eyes, I was ready to scream STOP!. In the latter case I no longer have any melanocytes where the burn took place and it took well over a month for the area to heal. This is better than cancer but not recommended unless it is needed.

In a lab setting I have had LN2 flow across my hand and also into one of my shoes twice that I can remember. A little of the liquid on the back of my hand was easy to shake off immediately. It just was a bit too cold. Luckily I could kick off the loafer type shoe quickly enough that there was no damage to my foot. Now I wear athletic, sneaker type shoes and I do not know if I could get one of them off quickly enough not to get a burn. Note that I have not been accidently burned in the last 25 years. One does get wiser with age!

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E - Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not represent the NIH. This message is not confidential and can be freely shared and reproduced.


________________________________
X-from: {jrminter-at-rochester.rr.com}
Reply-To: {jrminter-at-rochester.rr.com}

Dear List:

I seem to remember that one can dissolve "Scotch tape" in acetone (or
??) to make a glue for helping to adhere sections to grids.
Does anyone remember how much tape per unit of solvent?
Thanks!

Geoff

--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff-at-umdnj.edu
**********************************************



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From: guenter.resch-at-imba.oeaw.ac.at
Date: Thu, 19 Aug 2010 15:37:55 -0500
Subject: [Microscopy] Re: viaWWW: Safe handling of LN2 for EDS

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

Re: Palade, D.C. Histologic Techniques for Electron Microscopy, 2d Ed., 1964, Academic Press, NY, NY., p. 208.
"Unfortunately, carbon films do not adhere well to copper grids. However, it is possible to precoat the grid wires with an adhesive that helps. This author has, from time to time, used an extremely dilute solution of rubber cement in carbon tetrachloride. Or, the adhesive on a short length (about 4 in.) of cellophane tape, can be dissolved in an ounce of chloroform. The grids should be dipped in this and quickly dried on filter paper. The adhjisive must be so dilute that there is no tendency for it to form films or strands across the grid openings."

[I have seen this recipe using xylene/toluene on double-stick tape by immersing the tape and removing it after some time(?) - but I cannot remember the source.]

Hope this helps,

Fred Monson

Frederick C. Monson, PhD
Technical Director, CMIRT
West Chester University
West Chester, PA, 19383
610-738-0437
New Web Site:           http://cmirt.wcupa.edu/index.html


-----Original Message-----
X-from: mcauliff-at-umdnj.edu [mailto:mcauliff-at-umdnj.edu]
Sent: Thursday, August 19, 2010 1:53 PM
To: Monson, Frederick

Hi everyone,

while I agree with most things that were pointed out by Stephane and the
others, I was a little surprised by the following:

} Don't use closed shoes, I would recommend the
} ones with a holy upper part (I had to struggle to get these shoes
} because the chemists want closed shoes, which make sense when one
} works with chemicals or radioactivity)

I would recommend to do just the contrary when working with LN2: using
closed instead of holey shoes. The problem with holes on top is that
LN2 can enter, getting trapped in the shoe and causing serious burns as
prolonged exposure is guaranteed. As Pat already pointed out, this is
also the case for all kinds of sports shoes, that have some kind of
"mesh" on top, that is very easily penetrated by LN2.

As mentioned earlier, wearing boots with the trouser legs inside is not
a very good idea, either ...

By the way, there is a very nice poster from Leica Microsystems that
summarizes all those things ... maybe I can get permission to post it
somewhere.

Best,

Guenter

--
Dr. Guenter Resch, IMP-IMBA-GMI Electron Microscopy Facility
Email: guenter.resch-at-imba.oeaw.ac.at; Web: http://cores.imp.ac.at/em
Phone: +43 (1) 79044-4250; Fax/Voice Mail: +43 (1) 79044-224250
Post: Dr. Bohr-Gasse 3, 1030 Vienna, Austria, European Union

==============================Original Headers==============================
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From: kenconverse-at-qualityimages.biz
Date: Thu, 19 Aug 2010 16:07:19 -0500
Subject: [Microscopy] Re: viaWWW: Safe handling of LN2 for EDS

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

Guenter,
This thread seems to run every couple of years, so you can find earlier
threads in the archives. Generally speaking, the conclusion each time is
that one should handle lN2 in the nude so that it doesn't get trapped.
Somehow, I've never seen that posted in a lab even though it is the
considered opinion of many who handle the stuff frequently. ;-)

Actually, what bothers me the most is when people are required to use gloves
and what they are given are short, wide-cuffed "cryo" gloves that are a
funnel waiting to be used. Gloves with cuffs that go up to the elbow I
don't have such a problem with. Either way, one should be able to shake
them off instantly.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: guenter.resch-at-imba.oeaw.ac.at [mailto:guenter.resch-at-imba.oeaw.ac.at]
Sent: Thursday, August 19, 2010 4:42 PM
To: kenconverse-at-qualityimages.biz

Hi everyone,

while I agree with most things that were pointed out by Stephane and the
others, I was a little surprised by the following:

} Don't use closed shoes, I would recommend the
} ones with a holy upper part (I had to struggle to get these shoes
} because the chemists want closed shoes, which make sense when one
} works with chemicals or radioactivity)

I would recommend to do just the contrary when working with LN2: using
closed instead of holey shoes. The problem with holes on top is that
LN2 can enter, getting trapped in the shoe and causing serious burns as
prolonged exposure is guaranteed. As Pat already pointed out, this is
also the case for all kinds of sports shoes, that have some kind of
"mesh" on top, that is very easily penetrated by LN2.

As mentioned earlier, wearing boots with the trouser legs inside is not
a very good idea, either ...

By the way, there is a very nice poster from Leica Microsystems that
summarizes all those things ... maybe I can get permission to post it
somewhere.

Best,

Guenter

--
Dr. Guenter Resch, IMP-IMBA-GMI Electron Microscopy Facility
Email: guenter.resch-at-imba.oeaw.ac.at; Web: http://cores.imp.ac.at/em
Phone: +43 (1) 79044-4250; Fax/Voice Mail: +43 (1) 79044-224250
Post: Dr. Bohr-Gasse 3, 1030 Vienna, Austria, European Union

==============================Original Headers==============================
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From: jkrupp-at-deltacollege.edu
Date: Thu, 19 Aug 2010 17:16:52 -0500
Subject: [Microscopy] Sand

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

Hi

I am trying to get a lesson together for some local high schools. I am
using the LHS GEMS Microscopic Explorations for inspiration.

One of the exercises uses sand to tie in microscopy, geography and
observation. I have two samples of sand, one from Hawaii's North Shore
and maybe on or two from around the Santa Cruz, CA area.

Anybody have some from a collection to spare or willing to get some
and send it to me?

Thanks

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu




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From: vcsolomon-at-ysu.edu
Date: Thu, 19 Aug 2010 17:29:18 -0500
Subject: [Microscopy] viaWWW: Instrumentation Scientist Position

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Email: vcsolomon-at-ysu.edu
Name: Virgil C. Solomon

Organization: Youngstown State University

Title-Subject: [Filtered] Instrumentation Scientist Position

Message: Dear Colleagues,

In conjunction with the new Electron Microscopy Facility at
Youngstown State University, Youngstown, Ohio, we have an opening for
an Instrumentation Scientist. Below please find the position
description. More information and the on-line application (Posting
Number: 0600017) is available at https://jobs.ysu.edu.

Best regards,

Virgil C. Solomon

Position Description:

Maintenance, day-to-day operations, instruction (including
undergraduate students), and repair of equipment in the electron
microscopy facility including TEM and FIB/SEM instruments and sample
preparation laboratory. Instrumentation scientist will also provide
research support for internal and external EM users.
Operate and maintain the transmission electron microscope, focused
ion beam/scanning electron microscope and associated sample
preparation laboratory. Prepare TEM samples, primarily for samples
such as advanced composite materials.
Assist in the training of faculty, post-doctoral fellows,
undergraduate and graduate students, and industrial users in the
proper use of analytical equipment and instruments in the EM facility.
Assist researchers (both internal and external to YSU) and students
in the collection and interpretation of analytical data.
Maintain all necessary logs for the EM facility: operation, repair,
safety, etc.
Ensure adherence to University safety and chemical hygiene plans
where applicable.
Assist in the operation and maintenance of other analytical
instrumentation used for undergraduate laboratories in science and
engineering departments as needed.
Perform other related duties as assigned.

More information (Posting Number: 0600017) is available at https://jobs.ysu.edu



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From: schooley-at-mcn.org
Date: Thu, 19 Aug 2010 17:57:17 -0500
Subject: [Microscopy] Re: Sand

Contents Retrieved from Microscopy Listserver Archives
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} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America

MSA's Project MICRO has a sandpile, ably managed by Heidi Ullberg of
McCrone. You'll find it at
http://www.microscopy.org/education/projectMICRO/classroom.cfm

Caroline

--
Caroline Schooley
Project MICRO Coordinator
Microscopy Society of America
45301 Caspar Point Road, Box 117
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.microscopy.org/ProjectMICRO

==============================Original Headers==============================
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4, 17 -- To: jkrupp-at-deltacollege.edu, {Microscopy-at-microscopy.com}
4, 17 -- From: Caroline Schooley {schooley-at-mcn.org}
4, 17 -- Subject: Re: [Microscopy] Sand
4, 17 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed"
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From: jkrupp-at-deltacollege.edu
Date: Thu, 19 Aug 2010 18:10:26 -0500
Subject: [Microscopy] Fwd: Sand

Contents Retrieved from Microscopy Listserver Archives
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Begin forwarded message:

} From: Justin Kraft {kraftphysics-at-gmail.com}
} Date: August 19, 2010 3:30:08 PM PDT
} To: jkrupp-at-deltacollege.edu
} Subject: Re: [Microscopy] Sand
}
} Contact Caroline Schooley at the MSA. (schooley-at-mcn.org). She
} knows who maintains the sand supply for the MSA education
} department. They'll send you samples of sands from all over the
} world specifically for use in educational programs.
}
} --Justin A. Kraft



Indeed, I have been humbled. Here is Caroline's reply:

{MSA's Project MICRO has a sandpile, ably managed by Heidi Ullberg of
McCrone. You'll find it at

{http://www.microscopy.org/education/projectMICRO/SandCollection.cfm

Take a look at the list available, it will blow you away..

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu




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From: benada-at-biomed.cas.cz
Date: Fri, 20 Aug 2010 01:32:43 -0500
Subject: [Microscopy] Re: section adhesives for TEM grids

Contents Retrieved from Microscopy Listserver Archives
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Hello,
Here is the prescription from "Electron microscopy in molecular biology":

Dissolve the glue from 10 cm of adhesive tape in 50 ml of chloroform. Store
the solution in a dark bottle with tight-fitting stopper. ...

cit:
Sommerville, J.; Scheer, U., eds. Electron Microscopy in Molecular Biology: A
Practical Approach. Oxford: IRL Press, 1987. pp.147-148

Some comments:
We use Scotch Magic Tape and dissolve the glue from the tape for one or two
minutes. The rest of the tape is then removed with tweezers. Finally, the
"sticky solution" is filtered through dry filter paper.

Hope this works for you.

Best regards
Oldrich

--
Oldrich Benada
Institute of Microbiology
Acad. Sci. CR
Videnska 1024
CZ-142 20 Prague 4
Czech Republic


On Thursday 19 of August 2010 19:47:27 mcauliff-at-umdnj.edu wrote:
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} Dear List:
}
} I seem to remember that one can dissolve "Scotch tape" in acetone (or
} ??) to make a glue for helping to adhere sections to grids.
} Does anyone remember how much tape per unit of solvent?
} Thanks!
}
} Geoff

==============================Original Headers==============================
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From: nizets2-at-yahoo.com
Date: Fri, 20 Aug 2010 03:32:43 -0500
Subject: [Microscopy] section adhesives for TEM grids

Contents Retrieved from Microscopy Listserver Archives
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Hi!
 
This is a home-made recipe.
 
1. Take 15 cm tape as it is used by decorators and cut it in 1cm parts.
(Don't stick them together).
2. Fill a beaker with 15 ml choroform and place the 1cm-pieces inside under
stirring. Stirr for 1 min.
3. Cover the beaker with Alu foil and keep it in the hood for 90 mins.
4. Decant the solution and centrifuge in a resistant tube (Falcon) at 1300 rpm
at a bench centrifuge to get cellulose cut off.
5. Take the supernatant with a glass pipette and transfer it in a glass for
storage.
6. Dip the grid shorrtly in the solution and let it dry while you hold it in
horizontal position.(Dilute the solution if it clogs the grid space)





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From: j.r.thorpe-at-sussex.ac.uk
Date: Fri, 20 Aug 2010 03:33:02 -0500
Subject: [Microscopy] Re: section adhesives for TEM grids

Contents Retrieved from Microscopy Listserver Archives
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Hi Geoff,
About 10cm in (absolute) EtOH works for me. Just shake for a few seconds
and you can remove the tape.
Cheers,
Jules

Dr. Julian R. Thorpe
(Office 2C9/Lab 2C11-13)
Electron Microscope Division,
The Sussex Centre for Advanced Microscopy,
John Maynard-Smith Building,
School of Life Sciences,
University of Sussex,
Falmer,
Brighton BN1 9QG
Tel.: ext +44 (0)1273 877585
int 7585

URLs:
(home)
http://www.sussex.ac.uk/biology/profile2686.html
(lab)
http://www.lifesci.susx.ac.uk/Home/Julian_Thorpe/cover.htm
(research)
http://www.lifesci.susx.ac.uk/Home/Julian_Thorpe/ad_cover.htm
--On 19 August 2010 12:52 -0500 mcauliff-at-umdnj.edu wrote:

}
}
}
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} Dear List:
}
} I seem to remember that one can dissolve "Scotch tape" in acetone (or
} ??) to make a glue for helping to adhere sections to grids.
} Does anyone remember how much tape per unit of solvent?
} Thanks!
}
} Geoff
}
} --
} --
} **********************************************
} Geoff McAuliffe, Ph.D.
} Neuroscience and Cell Biology
} Robert Wood Johnson Medical School
} 675 Hoes Lane, Piscataway, NJ 08854
} voice: (732)-235-4583
} mcauliff-at-umdnj.edu
} **********************************************
}
}
}
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From: j.r.thorpe-at-sussex.ac.uk
Date: Fri, 20 Aug 2010 03:34:38 -0500
Subject: [Microscopy] Re: section adhesives for TEM grids

Contents Retrieved from Microscopy Listserver Archives
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Sorry, I meant in chloroform!!

--On 19 August 2010 12:52 -0500 mcauliff-at-umdnj.edu wrote:

} microscopy-at-microscopy.com



Dr. Julian R. Thorpe
(Office 2C9/Lab 2C11-13)
Electron Microscope Division,
The Sussex Centre for Advanced Microscopy,
John Maynard-Smith Building,
School of Life Sciences,
University of Sussex,
Falmer,
Brighton BN1 9QG
Tel.: ext +44 (0)1273 877585
int 7585

URLs:
(home)
http://www.sussex.ac.uk/biology/profile2686.html
(lab)
http://www.lifesci.susx.ac.uk/Home/Julian_Thorpe/cover.htm
(research)
http://www.lifesci.susx.ac.uk/Home/Julian_Thorpe/ad_cover.htm

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From: lherault-at-bu.edu
Date: Fri, 20 Aug 2010 08:08:08 -0500
Subject: [Microscopy] Re: section adhesives for TEM grids

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

It would be a good idea to practice this using modern tape, unless you have
some original 60s vintage tape, of course. I'm pretty sure the glue
formulation, not to mention the tape itself has changed radically since
then.

Ron L

-----Original Message-----
X-from: benada-at-biomed.cas.cz [mailto:benada-at-biomed.cas.cz]
Sent: Friday, August 20, 2010 2:36 AM
To: lherault-at-bu.edu

Hello,
Here is the prescription from "Electron microscopy in molecular biology":

Dissolve the glue from 10 cm of adhesive tape in 50 ml of chloroform. Store
the solution in a dark bottle with tight-fitting stopper. ...

cit:
Sommerville, J.; Scheer, U., eds. Electron Microscopy in Molecular Biology:
A
Practical Approach. Oxford: IRL Press, 1987. pp.147-148

Some comments:
We use Scotch Magic Tape and dissolve the glue from the tape for one or two

minutes. The rest of the tape is then removed with tweezers. Finally, the
"sticky solution" is filtered through dry filter paper.

Hope this works for you.

Best regards
Oldrich

--
Oldrich Benada
Institute of Microbiology
Acad. Sci. CR
Videnska 1024
CZ-142 20 Prague 4
Czech Republic


On Thursday 19 of August 2010 19:47:27 mcauliff-at-umdnj.edu wrote:
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} Dear List:
}
} I seem to remember that one can dissolve "Scotch tape" in acetone (or
} ??) to make a glue for helping to adhere sections to grids.
} Does anyone remember how much tape per unit of solvent?
} Thanks!
}
} Geoff

==============================Original Headers==============================
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From: benada-at-biomed.cas.cz
Date: Fri, 20 Aug 2010 08:53:04 -0500
Subject: [Microscopy] Re: section adhesives for TEM grids

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

Hello,
I agree with you that glue formulation has changed. I am sorry for vague
specification of the tape. We are using quite recent brand of Scotch tape:
Green box, Scotch Magic No. 810 (invisible); 19mm x 33m.
It seems to work.

Best regards from Prague
Oldrich

On Friday 20 of August 2010 15:10:43 you wrote:
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} --------------------------------------------------------------------------
} --
}
} It would be a good idea to practice this using modern tape, unless you have
} some original 60s vintage tape, of course. I'm pretty sure the glue
} formulation, not to mention the tape itself has changed radically since
} then.
}
} Ron L
}
} -----Original Message-----
} X-from: benada-at-biomed.cas.cz [mailto:benada-at-biomed.cas.cz]
} Sent: Friday, August 20, 2010 2:36 AM
} To: lherault-at-bu.edu
} Subject: [Microscopy] Re: section adhesives for TEM grids
}
}
}
}
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} --------------------------------------------------------------------------
} --
}
} Hello,
} Here is the prescription from "Electron microscopy in molecular biology":
}
} Dissolve the glue from 10 cm of adhesive tape in 50 ml of chloroform. Store
} the solution in a dark bottle with tight-fitting stopper. ...
}
} cit:
} Sommerville, J.; Scheer, U., eds. Electron Microscopy in Molecular Biology:
} A
} Practical Approach. Oxford: IRL Press, 1987. pp.147-148
}
} Some comments:
} We use Scotch Magic Tape and dissolve the glue from the tape for one or
} two
}
} minutes. The rest of the tape is then removed with tweezers. Finally, the
} "sticky solution" is filtered through dry filter paper.
}
} Hope this works for you.
}
} Best regards
} Oldrich
}
}
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}
} } - The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} } America To Subscribe/Unsubscribe --
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From: spurgeon-at-drexel.edu
Date: Fri, 20 Aug 2010 10:28:02 -0500
Subject: [Microscopy] TEM - Suggestions for wire saw

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

Hello everyone,

My group is interested in purchasing a diamond wire saw for sectioning
of metal and ceramic thin films (1-3 cm squares). We are considering
various options, such as Well (http://www.welldiamondwiresaws.com/)
and MTI (http://www.mtixtl.com/stx-602precisionwiresawwithsamplestageanddiamondwire-6diamaxsectioningcapability.aspx).
I would like to be able to rotate the sample in the x-y plane and
measure cuts precisely with a micrometer. Cost is a consideration, but
we are not averse to spending more for a model that will save us time
and frustration.

Those of you who use wire saws, what should we be looking for? Are
there any vendors or features you would recommend? Thank you for your
help.
--
Steven Spurgeon
Dynamic Characterization Group
Department of Materials Science & Engineering
Drexel University

Cell: 719.330.0441
Office: Bossone 102
Email: spurgeon-at-drexel.edu
Web: http://www.materials.drexel.edu/dcg/


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From: gw265-at-cam.ac.uk
Date: Sun, 22 Aug 2010 05:36:54 -0500
Subject: [Microscopy] slot grids, pioloform & formvar film

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

I'm doing a biological TEM project that involves a lot of serial fine
sectioning, so have to use slot grids (bar grids aren't practical
because of the number of sections and the size of the cells).

I am having difficulties with the film on my slot grids breaking under the
beam after about 10 minutes. In some cases it's possibly because the film
isn't really attached well to the grid in the first place.

Can anyone comment on how to make copper slot grids stick to pioloform or
formvar film? Is there a method that doesn't involve chloroform exposure?
Also, does carbon coating help to make film on slot grids more durable
under the beam?


Normally I would make pioloform in chloroform, cast films from slides that
have been washed in acetone, collect films (that show up uniformly pale
silver/gold) on metal rack holders (like a "formvar bridge" or "domino
rack" or whatever you call it
http://www.2spi.com/catalog/grids/domino.shtml); I wick the excess water
off with filter paper held gently at the edge of each hole on the
underside of the rack. At this stage I used to carbon coat the films
(back in the distant past when I had open access to someone to do the
carbon coating for me - I stopped doing this step when I moved labs to
somewhere that didn't carbon coat).

When sectioning, immediately prior to picking up sections, I dip cleaned
slot grids in the same solution of pioloform (to make them sticky) and
blow on them to get rid of any film forming across the slot. I pick up
the sections, then put the slot grid + wet sections down on the film. A
day or more later, I (gently) punch out the grids from the underside of
the film, pricking around the edges of the grid with fine forceps to
remove bits of film hanging off the sides of the grid.

If possible I'd like to change the method used for making grids & films
stick together, since by having to open the pot of chloroform/pioloform
solution repeatedly, I'm getting exposed to too much chloroform while
sectioning. This isn't an open invitation to debate health and safety
rules about chloroform: I, personally, would like to be exposed to less of
the stuff as it interferes massively with my ability to get work done.

thanks for any suggestions

Giselle

Dr Giselle Walker
University of Cambridge
UK



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12, 23 -- Subject: slot grids, pioloform & formvar film
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From: beth-at-plantbio.uga.edu
Date: Tue, 24 Aug 2010 10:14:17 -0500
Subject: [Microscopy] carbon-coating Formvar films

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

Hi all,
Has anyone successfully carbon-coated Formvar films using a sputter-
coater with a carbon filament?
I hear the carbon can be uneven and leave a chunky mess. If you have
refined this technique I'd like to hear from you.
The vacuum evaporators on campus are down at the moment so I am
seeking an alternative method.
thanks in advance for any advice,
Beth


==============================Original Headers==============================
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From: protrain-at-emcourses.com
Date: Tue, 24 Aug 2010 11:51:44 -0500
Subject: [Microscopy] carbon-coating Formvar films

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

Hi

Just a quick note about carbon film quality. Many years ago I worked with
Edwards high vacuum on a joint project where we tested all the different
vacuum methods for making a carbon film. One point stood out above all
others, the better the vacuum the better the film!

Run your sputter coater to try to obtain the highest vacuum possible prior
to coating.

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: beth-at-plantbio.uga.edu [mailto:beth-at-plantbio.uga.edu]
Sent: 24 August 2010 16:16
To: protrain-at-emcourses.com

Hi all,
Has anyone successfully carbon-coated Formvar films using a sputter-
coater with a carbon filament?
I hear the carbon can be uneven and leave a chunky mess. If you have
refined this technique I'd like to hear from you.
The vacuum evaporators on campus are down at the moment so I am
seeking an alternative method.
thanks in advance for any advice,
Beth


==============================Original Headers==============================
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==============================Original Headers==============================
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From: bozzola-at-siu.edu
Date: Tue, 24 Aug 2010 18:30:46 -0500
Subject: [Microscopy] EM: ultramicrotome repair

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

We are attempting to resurrect two older ultramicrotomes, a Sorvall
MT-2B and a Reichert Ultracut E, to help several graduate students in
their research projects.

The MT2B has a disintegrated drive belt that is essentially an O-ring.
Does anyone have information on how to change the belt?

The Ultracut may need professional servicing, so I am looking for a
reliable technical person to contact.

Isn't it amazing that these two instruments are still being used by so
many researchers! Our 40 year old LKB IIIs that are still cutting
away!

Cheers and thanks.
--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


==============================Original Headers==============================
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From: ngbrowning-at-ucdavis.edu
Date: Tue, 24 Aug 2010 19:24:00 -0500
Subject: [Microscopy] viaWWW: Facility Manager Position

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

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: ngbrowning-at-ucdavis.edu
Name: Nigel Browning

Organization: University of California, Davis

Title-Subject: [Filtered] Facility Manager Position

Message: Could you please post the following position? Thank you.

FACILITY MANAGER (82%-$40,000 annual)- start date to be determined.

Please submit curriculum vitae and list of
references via email to Dr. Nigel Browning,
nbrowning-at-ucdavis.edu, Department of Molecular
and Cellular Biology and the Department of
Chemical Engineering and Material Sciences

Job Summary:
Under the direction of the Facility Director,
provide day-to-day management and training for
three transmission electron microscopes within
the Interdisciplinary Center for Electron
Microscopy (ICEM). These three microscopes are
an integral part of the recharge-self supporting
ICEM facility and represent the microscopes in
the center that are primarily aimed at structural
biology. These microscopes are available to
registered and trained users on a 24 hour/7days a
week basis.

This position is a critical position and subject
to a background check. Employment is contingent
upon successful completion of a background
investigation including criminal history and
identity check. Non smoking work environment.
Work occasional overtime to meet operational needs

List positions supervised by this position:
None

JOB DESCRIPTION

45% A. Facility Management:

-Under the direction of the Director, provide
facility administration and management.
-Make recommendations in consultation with
customers of the Facility to the Facility
Director, and ICEM advisory committee, to assist
in determining long-and short-term goals for
operations.
-Ensure that the instruments are in safe working
condition at all times and that all users receive
safety and operatorís training. This will
include, but is not limited to, providing the
required safety training, documenting the
training, ensuring that users use safe work
practices; writing and maintaining a Statement of
Purpose (SOP) for each instrument and tool,
perform periodic safety inspections and
self-audits, and writing an annual safety report.
-Responsible for upgrades and improvement projects.
-Work closely with equipment service engineers to
reduce maintenance costs by trouble shooting
equipment issues prior to expensing costly
service visits.
-Maintain all facility related records,
instrument logbooks and service contracts.
-Develop user guides for users, reviewing and
updating equipment operating manuals
-Ensure that the user database is up-to-date.
-Provide monthly recharge activity.
-Assist the Management Services Officer (MSO)
with new and updated recharge rates
-Prepare statistical analytical reports to the
Facility Director, Department Chair and MSO and
Faculty Director when requested.
-Work closely with the MSO to maintain facility
budget, establish fiscal year budget closeout
reports, and project and anticipate future income
and expenses.
-Attend conferences/meetings, host new equipment
demonstrations and promote good working
relationships with users and outside vendors.
-Duties as assigned.

45% B. Technical and Research Support
-Responsible for the operation, maintenance, and
repair (routine and minor repairs) of the JEOL
1230, JEOL 2100 and JEOL 3000 TEMs. Also
responsible for maintaining standard shared
specimen preparation facilities for structural
biology. Back-up responsibilities for the other
four microscopes in the ICEM (primarily focused
on materials science), the aberration corrected
JEOL 2100, the JEOL 2500, the JEOL DTEM, and the
CM12. Also back-up responsibilities for shared
specimen preparation facilities for materials
science.
-Perform analyses and support users with one on one personal training.
-Responsible for supporting users with their
digital imaging, photoshop, and image and data
database analytical needs.
-Prepare reports and SOPs using word processors
and spreadsheets, and will use web-authoring
programs such as Microsoft Frontpage to maintain
and update content on the Facilityís web page.
-Interact and work closely with people from many
scientific disciplines, and from many different
cultures, and who have technical knowledge and
experience in electron microscopy ranging from
minimal to expert.
-When requested, assist users with the
preparation of instrumentation acquisition
grants, which utilize the electron microscopes.

10% C. Outreach
Serve as representative of the faculty by way of
incumbentís contact and interactions with
faculty, staff, researchers, and students, as
well as, with non-university clients,
particularly those from industry. Interact and
communicate with visitors from industry and
academia, international visitors and
representatives of professional societies on
operations, services, and equipment in the
Central Facility.

Skills, knowledge, and abilities, which are
essential for successful performance of this
position.

-Demonstrated knowledge with electron microscope
processing operations, including but not limited
to laboratory safety, hazardous waste storage and
disposal, electrical safety, safe use of cryogens
and compressed gases, and radiation safety.
-Advanced engineering experience and knowledge of
mechanical design practices to support research
projects/proposals.
-Degree in Engineering or an equivalent
combination of specialized education and training.
-Ability to work within the Principles of
Community; effective interpersonal communication
skills to work with staff and graduate students
in a laboratory environment.
-Skills in transmission electron microscopy
(TEM), TEM-based analyses and related sample
preparation techniques.
-Knowledge of the principles of operation of
vacuum pumps and gages, high-voltage supplies and
electron guns such as those employed in electron
microscopes, design and operation of cooling
systems, and the ability to make minor repairs
and conduct routine preventative maintenance of
electron microscopes.
-Skills in optical microscopy, including polarized and DIC techniques.
-Skills in digital darkroom techniques such as
image enhancement using Adobe Photoshop, basic
stereological techniques such as measuring size
and volume fraction, shapes, and orientation.
-Skills in electronics, such as the use of an
oscilloscope and multimeter, and basic
trouble-shooting.
-Skills in ultramicrotomy, staining, freeze-drying, and critical point drying.
-Skills in programming skills, such as Visual
BASIC, Delphi/Pascal, C++ or another high-level
programming language is preferred.
-Ability to develop protocols for procedures and
to train users in their application.
-Ability to articulate and work effectively with
others, including but not limited to
administrators, faculty, students, staff
(technical and administrative), researchers,
visiting scholars, non-university clients and
vendors.
-Ability to articulate the needs of the facility;
possess excellent written, oral and interpersonal
communication skills; excellent collaboration and
problem solving skills.
-Ability to set priorities and timelines for the
successful and timely completion of projects in a
cost effective manner. Strong organization
skills.




Login Host: 169.237.24.17
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From: oshel1pe-at-cmich.edu
Date: Wed, 25 Aug 2010 07:16:08 -0500
Subject: [Microscopy] Re: EM: ultramicrotome repair

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

John,

Try this guy:
MOC (Microscopical Optical Consulting Inc.) in Valley Cottage, NY,
for aftermarket microtome service! Thank You!

MOC
Helmut Patzig
Ph: 845-268-6450
Fax: 845-268-0172

Retired, I think, but still happy to help. An
MT-2 guru, used to service them when they were
The Microtome.
Patzig gave me this trick to replacing the drive
belt is *don't* remove anything so you can get
the old belt off and the new on on!
Cross-section the new belt - an o-ring, yes - at
45 degrees, thread through the pulleys, then
carefully superglue together the cut ends. Make
sure they meet properly and don't form a bump.
Hold until set, wait a bit, then start cutting.
"If it holds for 15 minutes, it will hold for
years."

Great machines. Work really well, stand up to
temporary moves to avoid construction and floods,
and annual undergrad TEM classes. Wish someone
would make them again. (Hey RMC - you bought the
Sorvall line, so that's a hint.)

The Ultracut I can't help with, sorry.

Phil

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

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576


==============================Original Headers==============================
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10, 28 -- To: bozzola-at-siu.edu
10, 28 -- From: Philip Oshel {oshel1pe-at-cmich.edu}
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From: kenconverse-at-qualityimages.biz
Date: Wed, 25 Aug 2010 07:32:35 -0500
Subject: [Microscopy] EM: ultramicrotome repair

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

Hi John,
Isn't it amazing how long and well something can work when there is no
computer involved? Of course it hasn't done a lot for sales, but why
consume when you don't have to. Keep that reliable vintage equipment
running!

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: bozzola-at-siu.edu [mailto:bozzola-at-siu.edu]
Sent: Tuesday, August 24, 2010 7:34 PM
To: kenconverse-at-qualityimages.biz

We are attempting to resurrect two older ultramicrotomes, a Sorvall
MT-2B and a Reichert Ultracut E, to help several graduate students in
their research projects.

The MT2B has a disintegrated drive belt that is essentially an O-ring.
Does anyone have information on how to change the belt?

The Ultracut may need professional servicing, so I am looking for a
reliable technical person to contact.

Isn't it amazing that these two instruments are still being used by so
many researchers! Our 40 year old LKB IIIs that are still cutting
away!

Cheers and thanks.
--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


==============================Original Headers==============================
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From: ehaller-at-health.usf.edu
Date: Wed, 25 Aug 2010 08:05:35 -0500
Subject: [Microscopy] Re: viaWWW: Ultracut S ultramicrotome controller

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

Hi, John,

I saved this message about Ultracuts from an earlier time on the listserver. Also, concerning MT-2's, contact Bill McGee, 7568 Florian Way, Liverpool, NY 13088. His number is 315-452-0828. He sells belt kits for MT-2's and 2-B's. The price for a 2-B kit last year was $35, part#16802. I have an MT2-B that finally gave up the ghost. It's giving me chatter problems that a belt refurb will not correct. I guess the lube in the unit went dry. I have the broken MT-2B, 2 working MT-2's, 3 MT-1's, and an LKB-3 up for grabs if anyone is interested ;) USF is moving me to new quarters, and I need to downsize. I'm getting new labspace, no new money for equipment, though :( , so I can't bring all of my microtomes with me.

Ed Haller

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
________________________________________
X-from: dac-at-research.umass.edu [dac-at-research.umass.edu]
Sent: Wednesday, June 23, 2010 7:45 AM
To: Haller, Edward

Peter,

If you don't get a donation and want to get it repaired, I have been
trying to connect people with aftermarket service for these older
microtomes and have a web page with some resources:

http://people.umass.edu/dac/projects/Open_Cut/Ultracut_Service.html

I recommend contacting MOC; Helmut Patzig formerly worked for Leica. He
knows people in Austria and has told me they can still supply the
controller boards - they aren't cheap, but neither is a new microtome.

I also have an "S" with dead controller. The main circuit board has
three separate microprocessors and memories of a now-obsolete types. The
functionality can today be provided by a $3 micro-controller; if only I
had more time....

Cheers!

Dale


Peter.Steele-at-allkids.org wrote:
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} Email: Peter.Steele-at-allkids.org
} Name: P. Steele
}
} Organization: All Children's Hospital
}
} Title-Subject: [Filtered] Ultracut S ultramicrotome controller.
}
} Message: I am looking for the Controller for a
} Reichert Ultracut S ultramicrotome. The machine
} is approximately 20 years old and Leica reports
} parts are no longer available for this workhorse.
} This is our backup ultramicrotome. The controller
} is defective but the ultramicrotome is in good
} shape.
}
}
} If you have an old Ultracut S ultramicrotome
} Controller that you are would like to donate
} please contact me offline.
}
} Thank you.
}
}
} Peter O. Steele, PhD, PMIAC, CLDir
} Pathology and Laboratory Medicine
} All Children's Hospital
} St. Petersburg, FL,
} USA, 33731-8920
}
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9, 20 -- From dac-at-research.umass.edu Wed Jun 23 06:38:34 2010
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From: oshel1pe-at-cmich.edu
Date: Wed, 25 Aug 2010 08:13:53 -0500
Subject: [Microscopy] Re: EM: ultramicrotome repair

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

John,

Ed Haller's email reminded me: there's also the
Open Cut web page for old microtomes.
http://people.umass.edu/dac/projects/Open_Cut/index.html

Dale Callaham runs this site, so I expect he will write in about it.

Phil

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(989) 774-3576


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From: oshel1pe-at-cmich.edu
Date: Wed, 25 Aug 2010 12:14:13 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist

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

Hi Beth

Been thinking a little more on your behalf. Use the type of string that is
like an old fashioned boot lace, it's almost a round tube. They normally
burn out in about 4 seconds so run for just 3; in the last second the string
throws carbon rocks at the specimen!

Hope this helps?

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: Beth Richardson [mailto:beth-at-plantbio.uga.edu]
Sent: 24 August 2010 18:50
To: protrain-at-emcourses.com

} Date: Tue, 24 Aug 2010 21:17:03 -0700 (PDT)
} From: Mohammed Tashkandi {tashkandi-at-gmail.com}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Tuesday, August 24,
} 2010 at 09:17:02 PM.
}
} realname - Mohammed Tashkandi
} Email - tashkandi-at-gmail.com
} ORGANIZATION - Colorado State University
} EDUCATION - Graduate College
} LOCATION - Fort Collins, Co, USA
} SUBJECT_OF_QUESTION - SEM images of CdS deposited on TCO glass
} QUESTION - Hi,
} I am trying to characterize some CdS thin film deposited on a
} transparent conductive oxide (TCO) coated glass as a part of CdTe
} thin film solar cells. I am using 30keV and 30,000x magnification.
} The SEM we have at CSU uses a secondary electron detector. The
} composition of the TCO is SnO2:F and its thickness according to the
} manufacturer is around 5000 angstroms.
This is a part of an effort to visualize discontinuities in the CdS
thin film. I was able to see some black spots under SEM and I am
trying to confirm that these are actual discontinuities. The CdS
regions look as bright white and I could see individual grains. Upon
moving to the dark spots, nothing seem to appear even if I try to
increase magnification or adjust brightness and contrast. One would
expect to see TCO grains if these black regions are actual
discontinuities but I could not see anything, just black regions with
no details or whatsoever. I am wondering whether there is a reason
why I could not see TCO grains in these black regions, is there any
issue regarding having two conductive layers on top of each other so
that what's beneath would not appear? I have not coated my samples
with gold since both CdS and TCO are conductive. We appreciate your
help and cooperation.
} Best regards.

--
***************************************************************************************
Forwarded from "Ask a Microscopist"
Please remember that the person asking the question is likely
not a member the listserver, and
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From: dsherman-at-purdue.edu
Date: Wed, 25 Aug 2010 15:32:40 -0500
Subject: [Microscopy] Determining wavelength for kVs

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Hi all,

There is a relatively simple formula for determining wavelength of x-rays
when the kinetic energy (KeV) is known. This is simply the wavelength =
12.3983 divided by the energy (KeV).

Is there a similar simple formula to determine wavelength of electrons
accelerated at a specific kV? I know that DeBroglie's formula of:
wavelength = Planck's constant divided by the mass of the electron x kV

should work but obviously I am confused by the different units. Math is
certainly not my strong suit! I just want to use this in examples when
explaining the Abbe equation and how wavelength relates to resolution.

Debby
---
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
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From: zaluzec-at-aaem.amc.anl.gov
Date: Wed, 25 Aug 2010 16:04:48 -0500
Subject: [Microscopy] Re: Determining wavelength for kVs

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Debby

Try this site it has a Java Tool for calculating what you asked for.

http://tpm.amc.anl.gov/NJZTools/EnergytoLambda.html

Nestor
Your Friendly Neighborhood SysOp



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From: bigelow-at-umich.edu
Date: Wed, 25 Aug 2010 17:38:01 -0500
Subject: [Microscopy] Re: Wavelength vs kV

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There is no really simple equation for
calculating the wavelength of electrons, because
for the accelerating voltages used in most TEMS
the electrons travel at a significant fraction of
the speed of light, and so their mass must be
corrected for the relativistic change. Thus we
have to contend with the relatively complicated
equation:

l =[SqRt (149.9/V)/SqRt(1 + 9.782 x 10-7 V)]x(1.002 x 10-8) cm


In this equation "SqRt" means take the square
root of the quantities in the following
parentheses (), the accelerating voltage is to
be expressed in Volts, not kilovolts, and the
answer comes out in centimeters. Thus, the
wavelength of electrons accelerated by 100 kV
(100,000 Volts) comes out to be 0.037 Å. For
accelerating voltages below about 20 kV
relativistic effects can be neglected and the
wavelength calculated approximately by SqRt(150/V)
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731


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From: chou_msa-at-yahoo.com
Date: Wed, 25 Aug 2010 18:28:49 -0500
Subject: [Microscopy] viaWWW: cut Silica glass using FIB

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Email: chou_msa-at-yahoo.com
Name: Alex Chou

Organization: Stevens Institute of Technology

Title-Subject: [Filtered] cut Silica glass using FIB

Message: Any suggestion of cutting a 30um thick Silica glass that has
been coated on a Si wafer using the FIB?

Thanks in advance,

Alex

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From: gunter.moeller-at-arkema.com
Date: Wed, 25 Aug 2010 18:29:13 -0500
Subject: [Microscopy] viaWWW: 2D Stylus Profilometer measurements

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Title-Subject: [Filtered] 2D Stylus Profilometer measurements

Message: We have a need to do stylus profilometer measurements
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Does anybody know a lab that can do such experiments (for a fee).

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Gunter Moeller, Arkema Inc.

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From: amgusman-at-ucdavis.edu
Date: Wed, 25 Aug 2010 18:29:39 -0500
Subject: [Microscopy] viaWWW: JEOL Service Technicians

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Name: Andrea Gusman

Organization: University of California, Davis

Title-Subject: [Filtered] JEOL Service Technicians

Message: Hi,

I was wondering if there is anyone that may have experience working
on older JEOL SEMs that are near the Davis, CA area?
We have a JEOL 848A SEM that is having image display problems.

The CRT screen and power supply appear to be in working order and we
currently do not have a budget to bring a service technician from
JEOL out to look at it to figure out the problem.

Thanks!
-Andrea

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From: larry.ackerman-at-ucsf.edu
Date: Wed, 25 Aug 2010 18:48:08 -0500
Subject: [Microscopy] Re: viaWWW: JEOL Service Technicians

Contents Retrieved from Microscopy Listserver Archives
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Contact Steve Kuzmic to help you with your JEOl SEM

Steve Kuzmic {kuzmics-at-sbcglobal.net}

Larry

amgusman-at-ucdavis.edu wrote:
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} Name: Andrea Gusman
}
} Organization: University of California, Davis
}
} Title-Subject: [Filtered] JEOL Service Technicians
}
} Message: Hi,
}
} I was wondering if there is anyone that may have experience working
} on older JEOL SEMs that are near the Davis, CA area?
} We have a JEOL 848A SEM that is having image display problems.
}
} The CRT screen and power supply appear to be in working order and we
} currently do not have a budget to bring a service technician from
} JEOL out to look at it to figure out the problem.
}
} Thanks!
} -Andrea
}
} Login Host: 169.237.55.116
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--
Larry Ackerman, Specialist
Electron Microscopy Lab
Manager, DERC Microscopy Core
UCSF, Dept. of Anatomy, Rm S1355
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

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From: Jerzy.Gazda-at-ceriumlabs.com
Date: Wed, 25 Aug 2010 19:49:06 -0500
Subject: [Microscopy] viaWWW: cut Silica glass using FIB

Contents Retrieved from Microscopy Listserver Archives
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Alex,
The thickness of the film will provide challenge with the milling time. Using enhancing etch chemistry can lead to speeding up the process. Depending on requirements for precision of your cut, and the size of the work-piece there might be several solutions:

1) IEEE etch to speed up removal of trench in SiO2 if the FIB cut is desired that is not much wider than the thickness (ie 30-50um cut)
2) If you need to produce a cross-section at a precise site, perhaps micro-cleaving the wafer piece to the site or nearby can lead to suitable result
3) There are fast speed ion milling tools that use hard-mask approach to produce cross-sections
4) Wafering saw could be used with micron-scale precisions to cut the film near your feature

Please contact me off-line to discuss those approaches in detail.

Hope this helps,

Jerzy


Jerzy Gazda

www.ceriumlabs.com

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Email: chou_msa-at-yahoo.com
Name: Alex Chou

Organization: Stevens Institute of Technology

Title-Subject: [Filtered] cut Silica glass using FIB

Message: Any suggestion of cutting a 30um thick Silica glass that has
been coated on a Si wafer using the FIB?

Thanks in advance,

Alex

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From: bigelow-at-umich.edu
Date: Wed, 25 Aug 2010 20:43:32 -0500
Subject: [Microscopy] Elect. Wavelengths

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

Since the question came up I thought I'd refresh my own memory, and
do something I haven't done for about 25 years: I calculated electron
wavelengths for several commonly-used accelerating voltages (using
Excel, of course). Here are the results:

5 kV 0.173 Angstroms
7 0.147
10 0.123
15 0.100
20 0.087
60 0.050
80 0.043
100 0.039
200 0.027
500 0.017
1000 0.012

Now, I feel almost like a graduate student, again.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From: bapinbhai-at-yahoo.co.in
Date: Thu, 26 Aug 2010 02:26:15 -0500
Subject: [Microscopy] Problem related to CrystalKitX and macTemPasX software

Contents Retrieved from Microscopy Listserver Archives
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In CrystalKitX I am making an interface of two crystal structures. Now it is not possible to save it as an executable file in MacTemPasX. In chapter 7 of CrystalKitX manual an example is shown where the void structure in InP was saved using "Write U. Cell to File" within File option (Page 41). But this option is inactive in our case (Version 1.9.5). If somebody is using the same software then please let me know the procudure to save an interface structure in CrystalKitX so that I can open it in MacTemPasX for image simulation.

Thank you.

Somnath Bhattacharyya





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From: bigelow-at-umich.edu
Date: Thu, 26 Aug 2010 04:08:02 -0500
Subject: [Microscopy] Wavelength MISTAKE!

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

I apologize, but I made a mistake in entering some of the arguments
for the functions used in my Excel spread sheet to calculate
electron wavelengths - I used a colon instead of a comma. As a
result, the values I gave in my previous E-mail are all off a bit.
Essentially, my mistakes cancelled the effect of the relativistic
correction. I corrected my spread sheet, and the correct value are:.

5 kV 0.1731 Angstroms (0.1732)
7 0.1461 (0.1464)
10 0.1221 (0.1225)
15 0.0994 (0.1000).
20 0.0859 (0.0866)
60 0.0487 (0.0500)
80 0.418 (0.0433)
100 0.370 (0.0387)
200 0.0251 (0.0274)
300 0.0197 (0.0224)
500 0.0142 (0.0173)
1000 0.0087 (0.0122)

The values in parentheses are the non-relativistic wavelengths
calculated by the simple formula SQRT(150/V). Essentially, it gives
pretty good values for accelerating voltages below about 10 kV, but
is rather far off for the higher voltages that are used in most TEMs.

My apologies, again, I am getting old!

--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From: protrain-at-emcourses.com
Date: Thu, 26 Aug 2010 05:17:23 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist

Contents Retrieved from Microscopy Listserver Archives
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Hi Mohammed

I am not familiar with your specimen but I may be able to help in relation
to general SEM operation.

Working at 30kV you are sampling a great deal of depth which may result in
information that is data piled upon data. May I suggest that changing the
accelerating voltage down in 5kV steps to section the specimen by kV. By
recording images of the same area this may provide you with a better
understanding of what is happening within your specimen.

If I were to do this with a client we would run a Monte Carlo simulation at
the same time to judge the information depth.

Good luck

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
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Sent: 25 August 2010 18:16
To: protrain-at-emcourses.com

} Date: Tue, 24 Aug 2010 21:17:03 -0700 (PDT)
} From: Mohammed Tashkandi {tashkandi-at-gmail.com}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Tuesday, August 24,
} 2010 at 09:17:02 PM.
}
} realname - Mohammed Tashkandi
} Email - tashkandi-at-gmail.com
} ORGANIZATION - Colorado State University
} EDUCATION - Graduate College
} LOCATION - Fort Collins, Co, USA
} SUBJECT_OF_QUESTION - SEM images of CdS deposited on TCO glass
} QUESTION - Hi,
} I am trying to characterize some CdS thin film deposited on a
} transparent conductive oxide (TCO) coated glass as a part of CdTe
} thin film solar cells. I am using 30keV and 30,000x magnification.
} The SEM we have at CSU uses a secondary electron detector. The
} composition of the TCO is SnO2:F and its thickness according to the
} manufacturer is around 5000 angstroms.
This is a part of an effort to visualize discontinuities in the CdS
thin film. I was able to see some black spots under SEM and I am
trying to confirm that these are actual discontinuities. The CdS
regions look as bright white and I could see individual grains. Upon
moving to the dark spots, nothing seem to appear even if I try to
increase magnification or adjust brightness and contrast. One would
expect to see TCO grains if these black regions are actual
discontinuities but I could not see anything, just black regions with
no details or whatsoever. I am wondering whether there is a reason
why I could not see TCO grains in these black regions, is there any
issue regarding having two conductive layers on top of each other so
that what's beneath would not appear? I have not coated my samples
with gold since both CdS and TCO are conductive. We appreciate your
help and cooperation.
} Best regards.

--
****************************************************************************
***********
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From: ehaller-at-health.usf.edu
Date: Thu, 26 Aug 2010 07:37:15 -0500
Subject: [Microscopy] I need a good protocol for TEM of C. elegans

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Dear Microscopists,

While on the subject of C. elegans, I have a researcher that wants to do studies on C. elegans at light level and TEM. I tried TEM of C. elegans before with dismal results, not getting good infiltration of fluids or plastic (acetone dehydration, epon equivalent plastic). Does anyone have a good protocol that they would care to share? Thanks in advance.

Ed


Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-cas.usf.edu

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From: jing.fu-at-monash.edu
Date: Thu, 26 Aug 2010 08:18:34 -0500
Subject: [Microscopy] viaWWW: Re:: cut Silica glass using FIB

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Email: jing.fu-at-monash.edu
Name: Jing Fu

Organization: Monash University

Title-Subject: [Filtered] Re:: cut Silica glass using FIB

Message: Alex,

1. You may have to a do thick coating if it does not affect your application.

2.For some platform/setup, you can fire the electron beam
simultaneously to neutralize the ibeam charging. The ebeam current
has to be 3-5 times of the ibeam. Some details:

D J Stokes et al 2007 J. Phys. D: Appl. Phys. 40 874

3. Some platform have gas needle based neutralizer.

Best of luck

Jing Fu

Login Host: 130.194.130.80
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From: somnath_tem-at-yahoo.com
Date: Thu, 26 Aug 2010 08:19:14 -0500
Subject: [Microscopy] viaWWW: Problem related to CrystalKitX and macTemPasX software

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Email: somnath_tem-at-yahoo.com
Name: Somnath Bhattacharyya

Organization: Tata Institute of Fundamental research

Title-Subject: [Filtered] Problem related to CrystalKitX and
macTemPasX software

Message: In CrystalKitX I am making an interface of two crystal structures.
Now it is not possible to save it as an executable file in MacTemPasX.
In chapter 7 of CrystalKitX manual an example is shown where the void
structure in InP was saved using "Write U. Cell to File" within File
option (Page 41). But this option is inactive in our case (Version
1.9.5).

If somebody is using the same software then please let me know the
procudure to save an interface structure in CrystalKitX so that I can
open it in MacTemPasX for image simulation.

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From: oshel1pe-at-cmich.edu
Date: Thu, 26 Aug 2010 10:49:13 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist

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I was wondering if anyone could recommend a good electron flight simulator program that will work on a newer, intel based mac without having to use something like parallels? I've found several programs that are written for the older classic mac version, or for a G4 CPU, but they won't work on a newer mac...

--Justin A. Kraft

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From anne-marie-at-crv.com Thu Aug 26 10:01:43 2010
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} Date: Thu, 26 Aug 2010 06:44:47 -0700 (PDT)
} From: Stephen Ruiz {stephen.ruiz-at-siemens.com}
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Thursday, August 26,
} 2010 at 06:44:45 AM.
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} SUBJECT_OF_QUESTION - Carbon Planchettes
} QUESTION - Ted Pella bought Ernest Fullum where I used to get highly
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} inferior. They sell a vitreous glassy carbon planchette for $70 ea.
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}
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From: ZZhang-at-uwyo.edu
Date: Thu, 26 Aug 2010 14:36:09 -0500
Subject: [Microscopy] I need a good protocol for TEM of C. elegans

Contents Retrieved from Microscopy Listserver Archives
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Hi Ed:

This might help -

Cohen M et al. Electron microscopy of lamin and the nuclear lamina in Caenorhabditis elegans. Methods Cell Biol. 2008;88:411-29.

http://www.ncbi.nlm.nih.gov/pubmed/18617045

Zhaojie

Zhaojie Zhang, Ph. D.
Director, Microscopy Core Facility
University of Wyoming
Laramie, WY 82071
PHONE: 307-766-3038
FAX: 307-766-5625




-----Original Message-----
X-from: ehaller-at-health.usf.edu [mailto:ehaller-at-health.usf.edu]
Sent: Thursday, August 26, 2010 6:42 AM
To: Z.J. Zhang

Dear Microscopists,

While on the subject of C. elegans, I have a researcher that wants to do studies on C. elegans at light level and TEM. I tried TEM of C. elegans before with dismal results, not getting good infiltration of fluids or plastic (acetone dehydration, epon equivalent plastic). Does anyone have a good protocol that they would care to share? Thanks in advance.

Ed


Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-cas.usf.edu

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From: jpare-at-emory.edy
Date: Thu, 26 Aug 2010 17:57:50 -0500
Subject: [Microscopy] viaWWW: Decommissioned Zeiss EM-10C

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Email: jpare-at-emory.edy
Name: Jeff ParÈ

Organization: Yerkes Research Center/Emory University

Title-Subject: [Filtered] Decommissioned Zeiss EM-10C

Message: Hi everybody,

We have an old EM-10C that we will decommission
in the next few months; It's in working order but
as you all know, that machine is no longer
supported by Zeiss anymore. We are looking for
someone who would be interested in taking it to
use it or for parts. It would cost you only the
scrap metal value and the moving costs...

Let me know if you are interested...I feel bad to
throw away a working instrument, but we need to
make place for the new EM.

Best regards

Jeff

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From: degraef-at-cmu.edu
Date: Thu, 26 Aug 2010 17:58:41 -0500
Subject: [Microscopy] viaWWW: post-doctoral opportunity at CMU

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Email: degraef-at-cmu.edu
Name: Marc DeGraef

Organization: Carnegie Mellon University

Title-Subject: [Filtered] post-doctoral opportunity at CMU

Message: Carnegie Mellon University
Department of Materials Science and Engineering

Position: Postdoctoral Research Associate

Description:
We have an immediate opening for a post-doctoral research associate
to work in the area of materials characterization using electron
microscopy methods, particularly transmission electron microscopy
(TEM), to understand local chemical and structural features in
functional ceramic coatings and heterostructures. The research
associate would work in close collaboration with industrial,
academic, and national laboratory partners in order to carry out
microstructural and compositional analysis of ceramic hard coatings
and solid-oxide fuel cells, with the goal of driving performance
improvements. The successful candidate should have a strong technical
background in materials characterization using various TEM/STEM/SEM
and related techniques, including energy dispersive spectroscopy
(EDS), selected-area electron diffraction (SAED), and high-resolution
TEM. The candidate must be able to work effectively as part of a
research team and have excellent verbal and written communication
skills. A strong publication record is preferred.

Qualifications:
Applicants must have a Ph.D. in physics, chemistry, materials
science, or a related field of study and extensive experience in
materials characterization by using advanced transmission electron
microscopy (TEM) and scanning TEM (STEM) techniques. Applicants
should be proficient in high-resolution chemical compositional
analysis using energy x-ray spectroscopy (EDS), and structural
analysis by HRTEM and SAED. Prior experience with electron energy
loss spectroscopy (EELS) is a plus. Experience in TEM sample
preparation by conventional methods, and particularly by focused ion
beam (FIB) lift-out, is critical. The applicant must have excellent
communication skills, a willingness to work in a collaborative
environment, and the ability to both generate new ideas and
independently pursue them. Experience in orientation imaging
microscopy (OIM) for grain size/orientation mapping in SEM, or
particularly by TEM, is preferred though not required.

Salary Range: Between $40,000-45,000/year

Appointment Duration: The position will have a one-year initial
appointment, with the possibility of extension.

How to Apply: Please send a cover letter, a CV, and a list of three
references to Prof. Yoosuf Picard at: ypicard-at-cmu.edu

Contact:
Yoosuf Picard
Assistant Research Professor
Department of Materials Science & Engineering
Carnegie Mellon University
Pittsburgh, PA


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From: W.Muss-at-salk.at
Date: Fri, 27 Aug 2010 01:20:18 -0500
Subject: [Microscopy] Re: I need a good protocol for TEM of C. elegans

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Good morning,
dear Zhaojie,
dear Ed, dear all,

this message just commenting the bibliographic hint given below:

The cited article

Cohen M et al. Electron microscopy of lamin and the nuclear lamina in Caenorhabditis elegans.
Methods Cell Biol. 2008;88:411-29
(Authors from: Division of Cell Biology, MRC-Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom,
European Molecular Biology Laboratory, Heidelberg, Germany; Department of Genetics, The Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel)

is Chapter 21 of/in:
Introduction to EM for Biologists ALLEN Terry D(ed), Methods in Cell Biology Series Vol.88 (2008), ACADEMIC PRESS Elsevier; Hardbound, 560 pages; Published: JUL-2008; ISBN 13: 978-0-12-374320-6
and costs ~ € 127.- incl. 10% tax (Europe) // ~ USD 156.00 (+ ?).
I own this book so perhaps I could share specific informations on that chapter if needed.

There is an other article by Cohen M et al (2002):

Transmission electron microscope studies of the nuclear envelope in Caenorhabditis elegans embryos.
Cohen M, Tzur YB, Neufeld E, Feinstein N, Delannoy MR, Wilson KL, Gruenbaum Y.
(Department of Genetics, The Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel)
Journal of Structural Biology 140, No 1-3 (Oct-Dec 2002) 232–240

which contains in the Materials & Methods section the following paragraph:

2.3. Sample preparation for TEM analysis:
Embryos were collected by the standard Clorox/M9 technique as described by Chen et al. (2000). A ~ 22 x 22mm square at or near the center of a Falcon 3001 petri dish (35 x 10mm) was marked. One drop (100–200µl) of 0.1% polylysine (Sigma P8920) was placed on the square and incubated for 5 min at room temperature and then the excess of polylysine was removed.
The petri dish was glued with double-stick tape to the lid of a 50-ml conical tube (petri dish facing up),
and washed and pelleted embryos in a drop of M9 buffer were placed on the polylysine-treated area of the petri
dish. The tubes were centrifuged at 1000g for 1 min in a swinging bucket. This step ensures that the embryos
remain on the polylysine for the rest of the procedure. The petri dish was removed from the tube, immediately
placed on ice, and given 1ml of fixative containing 1% freshly made formaldehyde, 2.5% EM grade glutaraldehyde
(Agar Scientific, Essex, UK), 0.1M Hepes, pH 7.0. The plate was sealed with parafilm and immediately
microwaved (Shef exclusive microwave oven SF-1875E at 70% microwave power) on ice for 20 s ON, 10 s OFF,
and then again 20 s ON. The sample was then cooled on ice for 5 min in a fume hood and the microwave cycle
was repeated once more. A Pelco-brand microwave, which provides a continuous power of 550 W, was also
used successfully. Fixation was completed by further incubation of the sample for 1 h at room temperature.
Excess fixative solution was then removed and the embryos were washed three times, 10 min each, with 1ml of
0.2M Hepes at pH 5.6. One milliliter of solution containing 1% OsO4, 0.5% reduced K4Fe(CN)6, 0.1M Hepes pH 5.6 was added, and the sample was incubated in the dark for 1 h. The sample was then washed twice, 10 min each, with ddH2O and then incubated for 10 min in 50% ethanol, 10 min in 70% ethanol, and 10 min in 90% ethanol, followed by 3 washes, 30 min each, in dry 100% ethanol. The sample was then incubated in a series of graded Epon/ethanol solutions, 2 h each, at room temperature with Epon/ethanol (1:2), Epon/ethanol (1:1), Epon/ethanol (2:1), overnight in fresh 100% Epon at 4° C in a desiccating chamber, and three more times, 2 h each, at room temperature with 100% Epon. The sample was embedded in Epon and polymerized for 2 days at 60 °C in a dry oven. The Epon block was then separated from the petri dish by soaking it in liquid N2.
The Epon block was sectioned to give thin sections, 80–90 nm, using a Diatome diamond knife. The sections
were picked up on 200 mesh thin bar copper grids and stained with uranyl acetate and lead citrate. Samples
were viewed with an electron microscope (Philips Technai 12) equipped with a MegaView II CCDcamera.
Following paragraph:

2.4. Immunogold labeling

.....

Hoping not to have counteracted with "copyright"-issues
Best wishes and regards


Wolfgang MUSS PhD
EM-Lab, Pathology, SALK-LKH (Gen. Hosp. SALZBURG)
Member of MSA since 1996 in good standing
SALZBURG, Austria



} -----Ursprüngliche Nachricht-----
} Von: ZZhang-at-uwyo.edu [mailto:ZZhang-at-uwyo.edu]
} Gesendet: Donnerstag, 26. August 2010 21:41
} An: Muß Wolfgang
} Betreff: [Microscopy] Re: I need a good protocol for TEM of C. elegans
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
} Hi Ed:
} This might help -
} Cohen M et al. Electron microscopy of lamin and the nuclear lamina in
} Caenorhabditis elegans. Methods Cell Biol. 2008;88:411-29.
} http://www.ncbi.nlm.nih.gov/pubmed/18617045

} Zhaojie
} Zhaojie Zhang, Ph. D.
} Director, Microscopy Core Facility
} University of Wyoming
} Laramie, WY 82071
} PHONE: 307-766-3038
} FAX: 307-766-5625


} -----Original Message-----
} X-from: ehaller-at-health.usf.edu [mailto:ehaller-at-health.usf.edu]
} Sent: Thursday, August 26, 2010 6:42 AM
} To: Z.J. Zhang
} Subject: [Microscopy] I need a good protocol for TEM of C. elegans
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} Dear Microscopists,
} While on the subject of C. elegans, I have a researcher that wants to do studies on C. elegans at light level and TEM. I tried TEM of C. elegans before with dismal results, not getting good infiltration of fluids or plastic (acetone dehydration, epon equivalent plastic). Does anyone have a good protocol that they would care to share? Thanks in advance.
} Ed
} Edward Haller, Lab Manager
} University of South Florida
} Integrative Biology Department
} Electron Microscopy Core
} SCA 110
} 4202 East Fowler Avenue
} Tampa, FL 33620
} 813-974-2676
} ehaller-at-cas.usf.edu
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From: ropope-at-gmail.com
Date: Fri, 27 Aug 2010 07:39:47 -0500
Subject: [Microscopy] viaWWW: JEOL 100S and ElectroScan 2020 for Redistribution

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Email: ropope-at-gmail.com
Name: Robert Pope

Organization: Indiana University South Bend

Title-Subject: [Filtered] JEOL 100S and ElectroScan 2020 for Redistribution

Message: I am posting this notice for my former institution. They
are remodeling the facility and need to find new homes for the
following microscopes. Please email them directly (email addresses
below), as I am not at the University any more.

Two Electron Microscopes Available for Redistribution


The Buyer needs to cover all costs of shipping the instruments to
their facility. It is possible to take only one of the instruments
if you do not need both. Please contact either Deb Marr
(dmarr-at-iusb.edu) or Andy Schnabel (aschnabe-at-iusb.edu).

(1) Transmission Electron Microscope JEOL JEM-100S
The TEM is a JEOL 100S TEM that was under service contract until 2004
and purchased by IU-South Bend in 2005. The TEM was under vacuum
until 2008. The TEM will need cleaning, all parts are included. It
still uses film, so it does not have digital capture. This
microscope was working in 2008, but has not been used since this
time. The TEM had blown a filament, but was keeping a stable vacuum.
The TEM and ESEM are hooked to a new Haskris chiller that runs both
instruments.

(2) Environmental Scanning Electron Microscope (ESEM) ElectroScan Model 2020
The SEM is the last model that ElectroScan made before being bought
out by Phillips, which was subsequently bought out by FEI. The ESEM
was under service contract until 2005, and under vacuum until 2008.
This microscope was also purchased in 2005. All parts are included.
It does have digital image capture. The ESEM was keeping a stable
vacuum, but the controller computer needed a new video card. Both
the TEM and ESEM are hooked to a new Haskris chiller that runs both
instruments.

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From: dsherman-at-purdue.edu
Date: Fri, 27 Aug 2010 14:08:26 -0500
Subject: [Microscopy] Tem prep for eye

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

Hi all you eye people,

We are having problems with drosophila eye prep. The investigator is
interested in the melanosomes and surrounding cell boundaries. She has
tried fixation with PAF-GLUT followed by Osmium and then embedded in Epon
generic resin after slow ~3day infiltration. We just do not have clear
membranes and the melanosomes tend to pop out during microtoming.

We were thinking of trying reduced osmium to try to get clearer membranes
(at low magnifications) and using Spurr's to try to get better infiltration.
We would appreciate alternative protocols from anyone with experience with
eyes.

Thanks,
Debby


---
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy




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From: chaueter-at-bcm.edu
Date: Tue, 31 Aug 2010 08:17:24 -0500
Subject: [Microscopy] Re: TEM Prep of Drosophila Eye

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

EFS might work using virtual PC. It works on WinXP.

gary g.


At 06:58 AM 8/26/2010, you wrote:



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From arzasia-at-iranscholarship.net Sat Aug 28 02:47:32 2010
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Email: bill-at-physics.uwa.edu.au
Name: andrew johnson

Organization: University of Western Australia

Title-Subject: [Filtered] want an old Gatan 636 sample holder

Message: If any one has an Gatan TEM double tilt cold holder,
designed about 1980, condition is not important, I would love to get
hold of it. Please contact me.

Many thanks, Andrew Johnson.

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From arabized1961-at-24protectplus123.com Mon Aug 30 07:21:48 2010
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Received: from [93.139.95.48] by mail1.vertrue.com; Mon, 30 Aug 2010 13:18:20 +0100

Dear Giselle,

I have struggled for around a year until I learned a protocol that
actually worked for me.


1.Prepare a formvar 1% solution in chlorophorm. Dilute overnight or with
low speed rotation for 30min. Protect from direct light exposure.

2.Do the protocol described by Peters, 2006. The link for the video is
http://www.currentprotocols.com/protocol/420/coating-grids-formvar-cb0407

(wow, a 2-step protocol it seems)

X-from my experience you will have to optimize the step when you let the
solution flow out of the burette.
For me it's working around 8s at constant speed to have silver to pearl
films.

I have been using this protocol for over a year with very good results.
You will not need carbon coating with this protocol but it may help the
grids to be usable for a longer time.

I hope this can help you as much as it helped me.
Please send some feedback if you eventually try it.

Best wishes,
Pedro Machado
Instituto Gulbenkian de Ciência
PT
}
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} I'm doing a biological TEM project that involves a lot of serial fine
} sectioning, so have to use slot grids (bar grids aren't practical
} because of the number of sections and the size of the cells).
}
} I am having difficulties with the film on my slot grids breaking under the
} beam after about 10 minutes. In some cases it's possibly because the film
} isn't really attached well to the grid in the first place.
}
} Can anyone comment on how to make copper slot grids stick to pioloform or
} formvar film? Is there a method that doesn't involve chloroform exposure?
} Also, does carbon coating help to make film on slot grids more durable
} under the beam?
}
}
} Normally I would make pioloform in chloroform, cast films from slides that
} have been washed in acetone, collect films (that show up uniformly pale
} silver/gold) on metal rack holders (like a "formvar bridge" or "domino
} rack" or whatever you call it
} http://www.2spi.com/catalog/grids/domino.shtml); I wick the excess water
} off with filter paper held gently at the edge of each hole on the
} underside of the rack. At this stage I used to carbon coat the films
} (back in the distant past when I had open access to someone to do the
} carbon coating for me - I stopped doing this step when I moved labs to
} somewhere that didn't carbon coat).
}
} When sectioning, immediately prior to picking up sections, I dip cleaned
} slot grids in the same solution of pioloform (to make them sticky) and
} blow on them to get rid of any film forming across the slot. I pick up
} the sections, then put the slot grid + wet sections down on the film. A
} day or more later, I (gently) punch out the grids from the underside of
} the film, pricking around the edges of the grid with fine forceps to
} remove bits of film hanging off the sides of the grid.
}
} If possible I'd like to change the method used for making grids & films
} stick together, since by having to open the pot of chloroform/pioloform
} solution repeatedly, I'm getting exposed to too much chloroform while
} sectioning. This isn't an open invitation to debate health and safety
} rules about chloroform: I, personally, would like to be exposed to less of
} the stuff as it interferes massively with my ability to get work done.
}
} thanks for any suggestions
}
} Giselle
}
} Dr Giselle Walker
} University of Cambridge
} UK
}
}
}
} ==============================Original
} Headers==============================
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12, 32 -- From pmachado-at-igc.gulbenkian.pt Mon Aug 30 12:08:10 2010
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From arteyparte-at-arteyparte.com Mon Aug 30 14:12:58 2010
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Hello Debby,

It could be fixative penetration issue. Our lab routinely processes adult fly eye for TEM.
We have used conventional and microwave irradiation (mwi) protocols with no problems.
Sometimes we encounter poorly preserved ultrastructure and it usually points out
to problems during dissection and fixation. After more careful dissection and fixation on
the next batch of samples, the problem is usually resolved.

To prevent penetration problems, especially at primary and secondary fixation,
either one of the eyes is slit open or the proboscis is carefully removed during dissection.
We place the fly heads in microcentrifuge tubes filled
with glut-paraformaldehyde-caco mix at 4 degree overnight on rotator.
The fly heads tend to float which is why we keep them on a rotator. Also, during the
secondary fixation step, we use tissue carriers that have open screen mesh on
both size (with small pore size) to keep the fly heads fully immersed during fixation steps.

Once the fly heads are osmium fixed, they no longer float.
We process them in microcentrifuge tubes and follow standard conventional (longer in fly tissues)
or mwi protocol (much faster approach).

We use embed 812 resin mix: Embed-812 25g, DDSA 12.5g, NMA 12.5 g, DMP-30 1.0 ml

Claire




Hi all you eye people,

We are having problems with drosophila eye prep. The investigator is
interested in the melanosomes and surrounding cell boundaries. She has
tried fixation with PAF-GLUT followed by Osmium and then embedded in Epon
generic resin after slow ~3day infiltration. We just do not have clear
membranes and the melanosomes tend to pop out during microtoming.

We were thinking of trying reduced osmium to try to get clearer membranes
(at low magnifications) and using Spurr's to try to get better infiltration.
We would appreciate alternative protocols from anyone with experience with
eyes.

Thanks,
Debby


---
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy





------------------------------------------------------------
Claire M. Haueter
Res. Tech. II (Electron Microscopy)
Bellen Lab
HHMI-Baylor College of Medicine
Mailstop BCM 235
One Baylor Plaza, Houston, TX 77030
Room T630
Phone: 713-798-8283
Fax: 713-798-3694
Email: chaueter-at-bcm.edu
-------------------------------------------------------------


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From: Cross-at-tru.ca
Date: Tue, 31 Aug 2010 15:35:00 -0500
Subject: [Microscopy] LM - Student Microscopes

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

Hello - my institution is looking to purchase student light compound microscopes. We are trying to decide between the Nikon E200 and the Olympus CX21.

If anyone out there has experience with these specific microscopes, either personally or via students, or comments about these two choices in general, I would be very grateful for your insights.


If you can help, please respond privately to me: cross-at-tru.ca


Thanks so much for your time,
Cindy


Cynthia Ross Friedman, Ph.D.
Associate Professor
Biological Sciences
Thompson Rivers University
Kamloops, British Columbia, Canada




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From: sbarlow-at-sunstroke.sdsu.edu
Date: Tue, 31 Aug 2010 16:46:44 -0500
Subject: [Microscopy] laser printers for lab

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

Dear all

Its time to replace my old printer with a newer one for general
confocal, SEM and TEM images around the lab, hand-outs, student
reports, etc.

Anyone have any suggestions? I am considering the HP color laserjet
CP4525dn or the Ricoh SP C430dn. Any others? I am avoiding the
inkjets due to my students running out multiples copies and devouring
my ink faster than I can refill them. The inkjets also don't lend
themselves to routine duplex printing....

Steve
--
Dr. Steven Barlow
EM Facility/Biology Dept.
San Diego State University
5500 Campanile Drive MC-4614
San Diego CA 92182-4614
phone: (619) 594-4523
fax: (619) 594-5676

email: sbarlow-at-sunstroke.sdsu.edu
http://www.sci.sdsu.edu/emfacility

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From: gary-at-gaugler.com
Date: Tue, 31 Aug 2010 19:52:27 -0500
Subject: [Microscopy] Re: laser printers for lab

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

I have a Ricoh C420dn and love it. I've had HP in the
past and did not care for it. I also had OKI LED color
printer that jammed all the time. The Ricoh has been solid.
Either color printer needs to have monitor color calibrated
and use Adobe gamut to get very close to WYSIWYG.

gary g.


At 02:48 PM 8/31/2010, you wrote:



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From: lisa-at-glosuntech.com
Date: Wed, 1 Sep 2010 01:01:05 -0500
Subject: [Microscopy] Denton 502 Carbon Jig Needed

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

Dear members,

Does anyone have used/spare Denton 502 Carbon Jig for sale? I need one to replace my broken carbon jig.


Lisa



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From: nyilmaz-at-mersin.edu.tr
Date: Wed, 1 Sep 2010 01:56:56 -0500
Subject: [Microscopy] advice for SEM

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

Dear Members,

We're planning to purchase a Low Vacuum SEM (JEOL 6510, Zeiss EVO or FEI
Quanta 450) for our central research facility. Is coating the any kind of
specimen (biological, material etc.) really unnecessary with a low vacuum
SEM. Shall we need a sputter coater and a critical point dryer with this
system? Any suggestion?
Thanks in advance...

Dr. Necat Yilmaz


__________ ESET NOD32 Antivirus Akýllý Güvenlik tarafýndan saðlanan bilgiler, virüs imza veritabaný sürümü: 5413 (20100831) __________

Ýleti ESET NOD32 Antivirus Akýllý Güvenlik tarafýndan denetlendi.

http://www.nod32.com.tr




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From: mike.bode-at-resaltatech.com
Date: Wed, 1 Sep 2010 09:13:35 -0500
Subject: [Microscopy] Re: laser printers for lab

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

Hi Gary,

As we are selling TEM cameras and SEM interfaces, we are constantly asked by
our customers about the best way to print the images they acquire, so I am
naturally interested in everybody's experience. Can you tell me what kind of
paper you use with the printer? In my experience the color laser printers
produce often a somewhat soft image with colors that are not very vibrant
compared to the colors you see on the screen. Perhaps not the most important
for EM images as they are mostly b/w, but I am wondering.

To anybody who is listening: If you want to send me a quick email with the
printer you are using with your EM camera or SEM interface, together with
perhaps a couple of sentences what you like or dislike about the printer and
how you achieve the best results, I could collect that information and
summarize it in a post here, provided I get enough information. Just send me
an email with the subject line "Printer poll", so I can sort those out
quickly. Thanks.


---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com





-----Original Message-----
X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com]
Sent: Tuesday, August 31, 2010 7:02 PM
To: mike.bode-at-resaltatech.com

I have a Ricoh C420dn and love it. I've had HP in the
past and did not care for it. I also had OKI LED color
printer that jammed all the time. The Ricoh has been solid.
Either color printer needs to have monitor color calibrated
and use Adobe gamut to get very close to WYSIWYG.

gary g.


At 02:48 PM 8/31/2010, you wrote:



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From: DusevichV-at-umkc.edu
Date: Wed, 1 Sep 2010 09:24:25 -0500
Subject: [Microscopy] RE: advice for SEM

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

I do not know about your research goals, but in general LV SEM does not replace high vacuum SEM in biological work. So, coating equipment is a must. CPD is desirable, but in some cases (but not always) it could be replaced with cheap treatment with HMDS. If you'll buy really LV (not ESEM) microscope, you cannot use it on hydrated specimens.

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

} -----Original Message-----
} From: nyilmaz-at-mersin.edu.tr [mailto:nyilmaz-at-mersin.edu.tr]
} Sent: Wednesday, September 01, 2010 1:57 AM
} To: Dusevich, Vladimir
} Subject: [Microscopy] advice for SEM
}
}
}
}
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} Dear Members,
}
} We're planning to purchase a Low Vacuum SEM (JEOL 6510, Zeiss EVO or
} FEI
} Quanta 450) for our central research facility. Is coating the any kind
} of
} specimen (biological, material etc.) really unnecessary with a low
} vacuum
} SEM. Shall we need a sputter coater and a critical point dryer with
} this
} system? Any suggestion?
} Thanks in advance...
}
} Dr. Necat Yilmaz
}
}
} __________ ESET NOD32 Antivirus Akýllý Güvenlik tarafýndan saðlanan
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From: donovan-at-uoregon.edu
Date: Wed, 1 Sep 2010 10:25:40 -0500
Subject: [Microscopy] advice for SEM

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

I have an FEI FEG Quanta 200 and I will say that
one of the things I like about it is its
tremendous flexibility in switching quickly
between high vacuum, low vacuum and E-sem modes.
The students like it because it is so easy to use.

To the question at hand I will say that coating
and high vacuum mode is generally desirable if it
is possible, but the low vacuum (with uncoated
samples) mode works quite well and we run
uncoated specimens all the time with excellent results.

But I agree with Vladimir that a coater is very
useful and for the relatively small cost should definitely be included.
john

At 07:27 AM 9/1/2010, DusevichV-at-umkc.edu wrote:



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From: wesaia-at-iastate.edu
Date: Wed, 1 Sep 2010 11:26:56 -0500
Subject: [Microscopy] advice for SEM

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

I concur witht he other couple replies I have seen so far.

Just because you can image a sample uncoated does not mean that you should (only) view it uncoated. You may wish to coat it and image it at high resolution. Therefore, you will probably want a coater. You might also try uncoated samples at low voltage, but the performance of that mode will depend on many things. Like I have seen many times on this list, it is best to try several modes and conditions rather than getting locked into only one way of doing things.

A quick example, I just had an investigator come by yesterday to review some images. He was calcining various compounds to produce CaO as a carbon dioxide sorbent. Since they were non-conductive particulates, I chose to examine them first in variable pressure mode using backscattered electron imaging. (Coated samples of this type still seem to charge a lot.) We found some interesting porosity. Now he wants to examine the samples again using secondary electrons on coated samples. One method does not always cover all needs.

For the record, we have a venerable Hitachi S-2460N which primarily operates in variable pressure mode with BSE. We also have a new FEI Quanta 250 which can operate in high vac, variable pressure, or environmental mode. It gets run in all modes.

Warren
________________________________________
X-from: nyilmaz-at-mersin.edu.tr [nyilmaz-at-mersin.edu.tr]
Sent: Wednesday, September 01, 2010 1:57 AM
To: wesaia-at-iastate.edu

Dear Members,

We're planning to purchase a Low Vacuum SEM (JEOL 6510, Zeiss EVO or FEI
Quanta 450) for our central research facility. Is coating the any kind of
specimen (biological, material etc.) really unnecessary with a low vacuum
SEM. Shall we need a sputter coater and a critical point dryer with this
system? Any suggestion?
Thanks in advance...

Dr. Necat Yilmaz


__________ ESET NOD32 Antivirus Akýllý Güvenlik tarafýndan saðlanan bilgiler, virüs imza veritabaný sürümü: 5413 (20100831) __________

Ýleti ESET NOD32 Antivirus Akýllý Güvenlik tarafýndan denetlendi.

http://www.nod32.com.tr




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From: greggps-at-umich.edu
Date: Wed, 1 Sep 2010 13:49:28 -0500
Subject: [Microscopy] Troubleshooting: TEM gun-Y drift. Need help with diagnosis.

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

Hello all,

Does anyone or has anyone had any luck or success of any kind running
an Olympus DP70 camera on 64 bit Windows 7? We would like to update
the computer operating the camera to Windows 7 64 bit, but have not
been able to locate any drivers. I have been able to find 64 bit
Vista drivers for the DP71 and 72, but no drivers past XP for the
DP70. I have contacted Olympus to see if they had any newer drivers
and have had no success. Any insight or help would be looked upon
kindly.

Mike
~~~~~~~~~~~~~~~~~~~~~
Michael Vollinger
Technical Assistant
Smith College
Dept. of Geosciences
Northampton, MA 01063
phone (413) 585-3765
fax (413) 585-3786
e-mail: mvolling-at-smith.edu


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From andrea-at-cotter-moroz.com Wed Sep 1 13:28:34 2010
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Our Philips CM100 BioTwin TEM has developed an electronic defect, which causes the gun Y-axis setting to drift. Some days the beam is relatively stable, some days you can see the beam 'jump' back and forth up to a centimeter's distance on the viewing stage (moving once every second), and some days we'll turn on the filament to find the beam completely missing, requiring a reasonable amount of alignment (not just the "Y" direction.)

Our service provider has not seen this previously, and has moved around and tested the circuit boards with no effect on the problem. His last two guesses are expensive (Probably $4,000-$7,000, and we've already put quite a bit into troubleshooting and power supplies this year), so I wanted to know if anyone else has encountered this phenomenon, and how you fixed it.

If we could confidently choose which item is defective, I can probably budget for it, but those holding the purse strings are a little nervous right now.

Your input is greatly appreciated. All suggestions or guesses are welcome.  
~Gregg

Gregg Sobocinski
Microscope Imaging Specialist
University of Michigan, MCDB Dept.
Ann Arbor, Michigan
USA




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From: FMonson-at-wcupa.edu
Date: Wed, 1 Sep 2010 15:09:50 -0500
Subject: [Microscopy] Re: advice for SEM

Contents Retrieved from Microscopy Listserver Archives
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Dear Dr. Yilmaz,
We have a Quanta 400 ESEM with Oxford INCA EDS with Feature/GSR, and we use what FEI calls "Low Vacuum"(up to 1 Torr) and "ESEM" (up- to 20 Torr with a Peltier cold stage) all the time.
The proper application of increased pressure, especially with water (gas/vapor) is to suppress charging on a specimen that is generally nonconductive. With this understanding, it is clear that there are a multitude of applications that can utilize variable pressures. A Peltier cold stage is the reason for success in the microscopy of biological specimens in temporary hydrated condition. [We have viewed - in ESEM mode - the effect of changing pressure at constant low temperature on the shapes of fungal spores.]
Coating (by evaporation or sputtering) and Critical Point drying (CPD) are accessory technolgies that have complementary uses in the applications that utilize variable pressure. "ESEM" and secondary electron modes (in variable pressure range) and various detectors that work and do not work through the range of pressures available in these instruments are also considerations that must be given attention.
Sputter coating at low thicknesses is most helpful in extracting higher signal from fine structural surface texture.
Our geologists can survey thin - or thick - sections of rock (95mm x 95mm x 10mm) that are NOT coated with carbon preliminary to selecting a region for thin sectioning or study of inclusions by a microprobe. [We have used the "Feature" module on Oxford INCA to survey for inclusions (monazites, zircons, and garnet crystals) useful in aging studies to the end that the geologist can choose from among hundreds of candidates whose data are stored in a database, relocate a few for better analysis before submitting even fewer for further study by EPMA. See the URL: http://cmirt.wcupa.edu/CMIRT_petrographic_thin_sections.html ]
My own attitude concerning the purchase of these instruments for academic institutions can be summed up by the word 'flexibility'. I always seek the best value that includes power, electronic stability and flexibility/breadth of use, coupled with the extended cost of service, which in these new instruments cannot be avoided - except at the peril of lost function and depressed user confidence.
I would suggest that a new microscope be viewed as a 'data mine' rather than an imaging device. The purchaser is always advised to seek the maximum analytic capabilities of such instruments. E.g., Get FEG if possible, and maximize the x-ray and imaging analytic power of the installation. Images are nice, but data are much, much better. AND, then there is the "Total Cost of Ownership". The most important advice is to have an honest estimate of extended cost as well as a realistic estimate of 'return' when you talk with deans and other administrators. They must be prepared to pay.

Hope this helps,

Fred Monson

Frederick C. Monson, PhD
Tecnhical Director
Center for Microanalysis and Imaging Research and Training Center (CMIRT)
Large Scientific Instrument Core
West Chester University S. Church St. and W. Rosedale Ave.
West Chester, PA, 19383
610-738-0437
fmonson-at-wcupa.edu

New Web Site: http://cmirt.wcupa.edu/index.html
New Scheduler: http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl
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From: stefan.diller-at-t-online.de
Date: Wed, 1 Sep 2010 15:38:36 -0500
Subject: [Microscopy] Re: Troubleshooting: TEM gun-Y drift. Need help with

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

Dear Gregg,
are you sure that it is an electronic problem?
Sounds like it might be some dirt in the inliner or wehnelt or somewhere in the gun.
Can you decrease the effect with decreasing the acceleration voltage and / or the condensor settings?

I would first do a thorough cleaning of the beam path.

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 01.09.10 20:54, schrieb greggps-at-umich.edu:
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} Our Philips CM100 BioTwin TEM has developed an electronic defect, which causes the gun Y-axis setting to drift. Some days the beam is relatively stable, some days you can see the beam 'jump' back and forth up to a centimeter's distance on the viewing stage (moving once every second), and some days we'll turn on the filament to find the beam completely missing, requiring a reasonable amount of alignment (not just the "Y" direction.)
}
} Our service provider has not seen this previously, and has moved around and tested the circuit boards with no effect on the problem. His last two guesses are expensive (Probably $4,000-$7,000, and we've already put quite a bit into troubleshooting and power supplies this year), so I wanted to know if anyone else has encountered this phenomenon, and how you fixed it.
}
} If we could confidently choose which item is defective, I can probably budget for it, but those holding the purse strings are a little nervous right now.
}
} Your input is greatly appreciated. All suggestions or guesses are welcome.
} ~Gregg
}
} Gregg Sobocinski
} Microscope Imaging Specialist
} University of Michigan, MCDB Dept.
} Ann Arbor, Michigan
} USA
}
}
}
}
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} 8, 29 -- Subject: Troubleshooting: TEM gun-Y drift. Need help with diagnosis.
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6, 23 -- From stefan.diller-at-t-online.de Wed Sep 1 15:38:35 2010
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From: eschumacher-at-mccrone.com
Date: Wed, 1 Sep 2010 16:00:08 -0500
Subject: [Microscopy] Short Course Announcement: TEM

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

Greetings Fellow Microscopists,

The Hooke College of Applied Sciences, located in Westmont, IL, is offering its TEM short course October 19-21, 2010. In addition to lectures, the course emphasizes hands-on training using state of the art equipment. For further details and registration information, please follow the link below:

http://www.hookecollege.com/

Regards,

Elaine

*********************************************************************
Elaine F.Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com



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From: gary-at-gaugler.com
Date: Wed, 1 Sep 2010 17:09:09 -0500
Subject: [Microscopy] Re: DP70 and Windows 7 64 bit

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

Win7 Pro has two modes: 32-bit x86 and 64-bit. You would
need Olympus DP SW 3.3 CD to install DP-70 in the x86
Program Files directory.

gary g.


At 10:42 AM 9/1/2010, you wrote:



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From: bozzola-at-siu.edu
Date: Wed, 1 Sep 2010 17:26:43 -0500
Subject: [Microscopy] EM: where to buy 50 mesh coated grids

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

Does anyone know where to purchase 50 mesh, nickel grids with Formvar coating?

I know, you are thinking, this guy is too lazy to make his own grids,
but we have been unable to get our films to strip from the slides for
a long time.

Thank you.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


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From: speransv-at-mail.nih.gov
Date: Wed, 1 Sep 2010 17:38:22 -0500
Subject: [Microscopy] EM: where to buy 50 mesh coated grids

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

I've been happy with Ladd Research and their Formvar+Light Carbon grids. The grids come freshly made (which does take an extra week) and hydrophilic.
And - each one is good.
The largest hole size on their website is 100 mesh, but I was told they could do custom work.

Another place where I inquired (2 years ago) and was told they will make any grid for you coated is SPI. I never got to compare, because their prices seemed higher.

Now, with the major EM supplier here - don't want to name names - their grids are the cheapest, but the quality is simply pathetic. With Ni in particular, you maybe get 10% useful grids, if you are lucky.

Vlad
________________________________________________
Vlad Speransky, Staff Scientist
Biomedical Engineering and Physical Science Shared Resource
National Institute of Biomedical Imaging and Bioengineering, NIH
13 South Dr, Rm. 3N17 MSC 5766
Bethesda, MD 20892
301 496-3989
vladislav_speransky-at-nih.gov
http://www.nibib.nih.gov/Research/Intramural/SharedResource/Speransky

Opinions and experiences related are those of Vlad Speransky and do not represent the NIH. On the good side, this message is not confidential and can be freely shared and reproduced.

________________________________________
X-from: bozzola-at-siu.edu [bozzola-at-siu.edu]
Sent: Wednesday, September 01, 2010 6:27 PM
To: Speransky, Vlad (NIH/NIBIB) [E]

Does anyone know where to purchase 50 mesh, nickel grids with Formvar coating?

I know, you are thinking, this guy is too lazy to make his own grids,
but we have been unable to get our films to strip from the slides for
a long time.

Thank you.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, ILÂ 62901
Phone: 618-453-3730


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From: rra-at-stowers.org
Date: Wed, 1 Sep 2010 18:06:37 -0500
Subject: [Microscopy] viaWWW: Ultramicrotomes

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Email: rra-at-stowers.org
Name: Rhonda Trimble

Organization: Stowers Institute

Title-Subject: [Filtered] Ultramicrotomes

Message: Dear All,

We are in the market for another ultramicrotome. I am interested in
finding out all that are available. I know there is Leica and RMC
(?). Does anyone know of any others? And can someone attest to
their preferences, and why?

Thanks so much in advance!

Rhonda

Rhonda Trimble HT(ASCP)HTL, QIHC
Electron Microscopy Specialist
Stowers Institute for Medical Research
1000 E 50th Street
Kansas City, Missouri 64110
816-926-4346
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From: wtivol-at-verizon.net
Date: Wed, 1 Sep 2010 18:33:24 -0500
Subject: [Microscopy] Re: EM: where to buy 50 mesh coated grids

Contents Retrieved from Microscopy Listserver Archives
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On Sep 1, 2010, at 3:33 PM, bozzola-at-siu.edu wrote:

} I know, you are thinking, this guy is too lazy to make his own grids,
} but we have been unable to get our films to strip from the slides for
} a long time.


Dear John,
Two problems you may be having are, first, humidity--the lower, the
better for films coming off--and, second, if you use nose grease on
the slides, the person's nose grease may not have the optimal
composition. My nose grease, for example, is iffy in that ordinary
films will come off, but holey films stay stubbornly stuck to the
slide. In the latter case, I dissolved a small amount of Apiazon L in
petroleum ether and coated the slides with that, then the holey films
came right off. I'm sorry I do not remember the amount of Apiazon I
used.
Yours,
Bill


==============================Original Headers==============================
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From: colijn.1-at-osu.edu
Date: Wed, 1 Sep 2010 19:54:29 -0500
Subject: [Microscopy] EM: where to buy 50 mesh coated grids

Contents Retrieved from Microscopy Listserver Archives
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Hi all,

Rather than casting the films onto glass slides, try using freshly
cleaved mica. I have seen many recipes (formvar voodoo) for
coating/polishing/treating glass slides, but have never had good success
casting films on them. If you score the edge of the mica, the water
penetrates easily and the films float right off.

If you insist on using glass slides, I would opt for a detergent based
coating rather than any kind of grease (nose or otherwise). The
detergent should give less contamination since it dissolves in the water
used for floating off the film. I've used standard lab detergent (no
lemon scent added!) for coating ion mill windows and bell jars with good
success. Dilute it a bit, coat the glass, then polish most of the
detergent off. The deposits usually come right off the glass with maybe
a little bit of rubbing.

After getting some silicone contaminated carbon films from a major EM
supply house, I decided that I need to make my own. They aren't as
pretty, but at least they're clean.

Cheers,
Henk



At 9/1/2010 7:34 PM, wtivol-at-verizon.net wrote:
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe --http://www.microscopy.com/MicroscopyListserver
} On-Line Helphttp://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
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}
} On Sep 1, 2010, at 3:33 PM,bozzola-at-siu.edu wrote:
}
} } I know, you are thinking, this guy is too lazy to make his own grids,
} } but we have been unable to get our films to strip from the slides for
} } a long time.
} Dear John,
} Two problems you may be having are, first, humidity--the lower, the
} better for films coming off--and, second, if you use nose grease on
} the slides, the person's nose grease may not have the optimal
} composition. My nose grease, for example, is iffy in that ordinary
} films will come off, but holey films stay stubbornly stuck to the
} slide. In the latter case, I dissolved a small amount of Apiazon L in
} petroleum ether and coated the slides with that, then the holey films
} came right off. I'm sorry I do not remember the amount of Apiazon I
} used.
} Yours,
} Bill
}
}
} ==============================Original Headers==============================
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}

--
Hendrik O. Colijn www.ceof.ohio-state.edu
OSU Campus Electron Optics Facility colijn.1-at-osu.edu
040 Fontana Labs (614) 292-0674 (V)
116 W. 19th Ave. (614) 292-7523 (F)
Columbus, OH 43210

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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From: bozzola-at-siu.edu
Date: Wed, 1 Sep 2010 22:14:25 -0500
Subject: [Microscopy] Re: EM: where to buy 50 mesh coated grids

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

We've tried everything others have mentioned -- except for the bad
breath. So, tomorrow, I shall consume a large number of onions and
garlic, washed down with a liberal amount of bourbon prior to
breathing onto the Formvar. If nothing else, it won't matter if the
film doesn't separate, since I will be oblivious.

J. Bozzola

On Wed, Sep 1, 2010 at 7:15 PM, Markus F. Meyenhofer
{micro-at-superlink.net} wrote:
} Your slides are too clean!!
} Just rub relative clean slides with KimWhipes, dip in formvar  cut the edges
} and breath  (heavy, and bad breath helps ;-)  , I always tell this to my
} students) on them before lowering into the water.
} Of course, I know, you know this already!
} Regards,
} Markus
} ----- Original Message ----- From: {bozzola-at-siu.edu}
} To: {micro-at-superlink.net}
} Sent: Wednesday, September 01, 2010 6:26 PM
} Subject: [Microscopy] EM: where to buy 50 mesh coated grids
}
}
} }
} }
} }
} }
} } ----------------------------------------------------------------------------
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} }
} } ----------------------------------------------------------------------------
} }
} } Does anyone know where to purchase 50 mesh, nickel grids with Formvar
} } coating?
} }
} } I know, you are thinking, this guy is too lazy to make his own grids,
} } but we have been unable to get our films to strip from the slides for
} } a long time.
} }
} } Thank you.
} }
} } --
} } John J. Bozzola, Ph.D., Director
} } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
} } Southern Illinois University


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From: paul_hazelton-at-umanitoba.ca
Date: Thu, 2 Sep 2010 00:04:17 -0500
Subject: [Microscopy] EM: where to buy 50 mesh coated grids

Contents Retrieved from Microscopy Listserver Archives
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John

Henk mentioned the nose grease thing on the list server a number of
years ago. Tried it. May work for him, it was a total flop for me.

We had trouble off and on for years. Super cleaned the slides, put
grease on them, etc. One thing I noticed is how recently the slides
were made seemed to have an effect. Invariably, we have trouble when
the box of slides is old. The second they start to go milky, have
whitish spots, etc, we cannot get the slides off. But slides from a new
box, recently purchased, consistently will release the film. The other
problem we've found is using slides that are super clean - acetone,
ethanol, etc. But as Markus said, a slide fresh out of the box, wiped
with a kimwipe works best (actually, I use a lab coat sleeve, but then
I've been accused of being some form of reprobate anyway).

The second thing is how long the formvar has been on the slide.
Actually tried this silly little experiment just because I got curious
as to whether I could make up a box of slides at one time, and use them
over the next several months. Dipped slides and tried floating the
formvar off after 10, 15, 30, 45, 60, 90, 120mins, 4, 8, 16, 24, and 48
hours. Did 10 slides each.

Any where from 10 to 30 minutes was great, after that slides started to
give a little trouble. About 75% of slides released the formvar after 2
hours. By 16 hours, almost no slides released the formvar. So making
the film on slides in advance, and casting them into the water days or
months later is just not going to work, at least in our hands.

Breathing on the slides. Yep, it works. Just think back and do your
Artie Johnson dirty old man bit from 'Laugh In' days. Actually, you
don't have to breath that heavily. And if the family and techs don't
mind, and you really like the escargots bourguignon, then go for it.
And sop up the butter with that chunk of baguette. Make the process of
making slides a celebratory excuse.

Also, what are you making them in. We use ethylene dichloride. Have
seen other solvents recommended, never tried them. Make it up 100ml at
a time, pour it back into the coplin jar after each use - now everyone
can scream about contamination, bad technique, etc - but it works. Be
practical, help limit the amount of pollution you put into the
environment, and stay out of the clutches of your Health and Safety
Office. We dip about 12 slides, and immediately pour the formvar back
into a bottle, seal with parafilm, and wait to use it later. I have
used it for up to 2 years. Having said that, I also always float off
the first film, put a single grid on it, and then check that grid in the
microscope immediately to make sure there are no problems with the
formvar solution before I make up 400-500 grids.

We are still making up over 3,000 grids a year. Since the tech that was
soooo good at making them became ill and passed away I have just made
them myself. Too much trouble getting the other techs to get it right.
I make up 1200 at a time, and make up a batch every time we get down
to 200. Good mindless way to waste a Monday morning. Never had a
problem with longevity - but then the grids never stay around more that
6 months at tops.

Final point. Someone once asked me if I didn't find Formvar to be weak
compared to collodion. A true gentleman and brilliant virologist -
suspect the question was a "Hazelton, hmm, heard about him, how bright
is he really" type of test. I made the observation then, as now, that
all plastics are essentially the same. If your films are weak, just
make a higher concentration solution next time. And if the lab you go
into insists one plastic is better than another, go with the flow. Not
worth the argument, and like politics and religion, you will never
change a primal belief.

And I've been so good the last several years, sticking to the
gastroenteric molecular work, not polluting the band width, etc.....
Next thing you know Phil's going to send me a note asking if I will put
it together as a technique thing for Microscopy Today, and I still owe
him something he asked me for several years ago, and cannot remember
what it was for the life of me.

Oh yeah, work avoidance, that's why I stopped polluting the band width....

Paul
--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 Bannatyne Avenue
Winnipeg, Manitoba, Canada, R3E 3J7
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926


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From: paul_hazelton-at-umanitoba.ca
Date: Thu, 2 Sep 2010 00:21:01 -0500
Subject: [Microscopy] Re: EM: where to buy 50 mesh coated grids - Redux

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

Sorry, it was Bill, not Henk with the Nose Grease thing.

Paul
--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 Bannatyne Avenue
Winnipeg, Manitoba, Canada, R3E 3J7
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926


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From: protrain-at-emcourses.com
Date: Thu, 2 Sep 2010 03:14:47 -0500
Subject: [Microscopy] Re: Troubleshooting: TEM gun-Y drift. Need help with

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

Hi

Well I believe Stefan has taken you in the correct direction.

This type of problem is typical of dirt in the column, it charges deflecting
the beam, then on discharge it allows the beam to fall back.

A simple test is to reduce the gun's emission/beam current which will reduce
the rate of charge, or allow a higher emission/beam current which will
increase the amount of charge.

We would also expect a change in charge rate with a change of kV, however
when you change the kV you change the deflection coil currents, so that
would not remove them from the equation!

With a problem further down the column changing spot size (1) and or
condenser aperture size (2), followed by a change in charge rate would be a
good indication of (1) dirt below condenser one, or (2) dirt below the
aperture position.

If you use resin embedded specimens the volatile components move into the
column moving from area to area until they eventually find a "cool" position
where they sit without problems. As time moves on the contamination in this
area builds to a point when charge discharge occurs. This problem area is
usually just above the specimen in the deflection coil liner. Unfortunately
simply cleaning this area just polishes the resin so quite brutal techniques
are required.

Try the tests and then educate your service staff that unstable deflection
coils are very rarely the problem; never in my 44 years!

Good luck

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: stefan.diller-at-t-online.de [mailto:stefan.diller-at-t-online.de]
Sent: 01 September 2010 21:40
To: protrain-at-emcourses.com

Dear Gregg,
are you sure that it is an electronic problem?
Sounds like it might be some dirt in the inliner or wehnelt or somewhere in
the gun.
Can you decrease the effect with decreasing the acceleration voltage and /
or the condensor settings?

I would first do a thorough cleaning of the beam path.

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 01.09.10 20:54, schrieb greggps-at-umich.edu:
}
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} Our Philips CM100 BioTwin TEM has developed an electronic defect, which
causes the gun Y-axis setting to drift. Some days the beam is relatively
stable, some days you can see the beam 'jump' back and forth up to a
centimeter's distance on the viewing stage (moving once every second), and
some days we'll turn on the filament to find the beam completely missing,
requiring a reasonable amount of alignment (not just the "Y" direction.)
}
} Our service provider has not seen this previously, and has moved around
and tested the circuit boards with no effect on the problem. His last two
guesses are expensive (Probably $4,000-$7,000, and we've already put quite a
bit into troubleshooting and power supplies this year), so I wanted to know
if anyone else has encountered this phenomenon, and how you fixed it.
}
} If we could confidently choose which item is defective, I can probably
budget for it, but those holding the purse strings are a little nervous
right now.
}
} Your input is greatly appreciated. All suggestions or guesses are welcome.
} ~Gregg
}
} Gregg Sobocinski
} Microscope Imaging Specialist
} University of Michigan, MCDB Dept.
} Ann Arbor, Michigan
} USA
}
}
}
}
} ==============================Original
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} 8, 29 -- Subject: Troubleshooting: TEM gun-Y drift. Need help with
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From: nizets2-at-yahoo.com
Date: Thu, 2 Sep 2010 03:19:24 -0500
Subject: [Microscopy] Re: Troubleshooting: TEM gun-Y drift. Need help with

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

I would tend to think the same way as Stefan, and not only because we have the
same first name :-)
It reminds me strongly of a recent and similar problem on the list with a SEM,
where contamination of the column was pointed out.

Regards,

Stephane


 


----- Original Message ----
X-from: "stefan.diller-at-t-online.de" {stefan.diller-at-t-online.de}
To: nizets2-at-yahoo.com
Sent: Wed, September 1, 2010 10:42:21 PM

Dear Gregg,
are you sure that it is an electronic problem?
Sounds like it might be some dirt in the inliner or wehnelt or somewhere in the
gun.
Can you decrease the effect with decreasing the acceleration voltage and / or
the condensor settings?

I would first do a thorough cleaning of the beam path.

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 01.09.10 20:54, schrieb greggps-at-umich.edu:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Our Philips CM100 BioTwin TEM has developed an electronic defect, which causes
} the gun Y-axis setting to drift. Some days the beam is relatively stable, some
} days you can see the beam 'jump' back and forth up to a centimeter's distance on
} the viewing stage (moving once every second), and some days we'll turn on the
} filament to find the beam completely missing, requiring a reasonable amount of
} alignment (not just the "Y" direction.)
}
} Our service provider has not seen this previously, and has moved around and
} tested the circuit boards with no effect on the problem. His last two guesses
} are expensive (Probably $4,000-$7,000, and we've already put quite a bit into
} troubleshooting and power supplies this year), so I wanted to know if anyone
} else has encountered this phenomenon, and how you fixed it.
}
} If we could confidently choose which item is defective, I can probably budget
} for it, but those holding the purse strings are a little nervous right now.
}
} Your input is greatly appreciated. All suggestions or guesses are welcome.
} ~Gregg
}
} Gregg Sobocinski
} Microscope Imaging Specialist
} University of Michigan, MCDB Dept.
} Ann Arbor, Michigan
} USA
}
}
}
}
} ==============================Original Headers==============================
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} 8, 29 -- To: Microscopy MSA {Microscopy-at-microscopy.com}
} 8, 29 -- Date: Wed, 1 Sep 2010 14:49:23 -0400
} 8, 29 -- Subject: Troubleshooting: TEM gun-Y drift. Need help with diagnosis.
} 8, 29 -- Thread-Topic: Troubleshooting: TEM gun-Y drift. Need help with
} diagnosis.
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From: oshel1pe-at-cmich.edu
Date: Thu, 2 Sep 2010 08:03:34 -0500
Subject: [Microscopy] Re: EM: where to buy 50 mesh coated grids

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

John,

Not a source of 50 mesh grids, but ... what slides are you using?
We find that we have to use the really cheap
P11011 generic chinese slides to get consistent
release. These are bought in bulk for the Intro
Bio course from VWR (I think - I'd have to ask
the Intro Bio guy). Anything else is too good a
quality to release well, even with the nose trick.
The PacSci P11011 slides should work, but the
cheapie Bellweather brand slides don't release.

Phil

} Does anyone know where to purchase 50 mesh, nickel grids with Formvar coating?
}
} I know, you are thinking, this guy is too lazy to make his own grids,
} but we have been unable to get our films to strip from the slides for
} a long time.
}
} Thank you.
}
} --
} John J. Bozzola, Ph.D., Director
} I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
} Southern Illinois University
} 750 Communications Drive
} Carbondale, IL¬Ý 62901
} Phone: 618-453-3730

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576


==============================Original Headers==============================
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6, 28 -- To: bozzola-at-siu.edu
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From: gregory.a.jerman-at-nasa.gov
Date: Thu, 2 Sep 2010 08:10:11 -0500
Subject: [Microscopy] Re: DP70 and Windows 7 64 bit

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

Windows 7 64-bit supports virtualization of the 32-bit WinXP environment
under certain circumstances. First you must use the Professional, Ultimate,
or Enterprise editions. You then have to have a computer that supports
hardware virtualization. Many older Intel CPU's do not support
virtualization, so having the correct operating system won't matter. The
easiest way to check is to get into the computer BIOS screen and see if you
can turn on virtualization under the CPU parameters. If not, then your
computer doesn't support it. Once you enable CPU virtualization, you need
to download Windows XP Mode software and Windows Virtual PC software from
Microsoft. Once the virtual WinXP environment is setup, you should then be
able to install the 32-bit WinXP drivers and camara software just like they
were being installed on a true WinXP system. There are no guarantees it
will work, but those are the hoops you will have to jump through.

Good Luck,

Greg



} From: "gary-at-gaugler.com" {gary-at-gaugler.com}
} Reply-To: "gary-at-gaugler.com" {gary-at-gaugler.com}
} Date: Wed, 1 Sep 2010 17:12:53 -0500
} To: "Jerman, Gregory A. (MSFC-EM31)" {gregory.a.jerman-at-nasa.gov}
} Subject: [Microscopy] Re: DP70 and Windows 7 64 bit
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Win7 Pro has two modes: 32-bit x86 and 64-bit. You would
} need Olympus DP SW 3.3 CD to install DP-70 in the x86
} Program Files directory.
}
} gary g.
}
}
} At 10:42 AM 9/1/2010, you wrote:
}
}
}
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } Hello all,
} }
} } Does anyone or has anyone had any luck or success of any kind running
} } an Olympus DP70 camera on 64 bit Windows 7? We would like to update
} } the computer operating the camera to Windows 7 64 bit, but have not
} } been able to locate any drivers. I have been able to find 64 bit
} } Vista drivers for the DP71 and 72, but no drivers past XP for the
} } DP70. I have contacted Olympus to see if they had any newer drivers
} } and have had no success. Any insight or help would be looked upon
} } kindly.
} }
} } Mike
} } ~~~~~~~~~~~~~~~~~~~~~
} } Michael Vollinger
} } Technical Assistant
} } Smith College
} } Dept. of Geosciences
} } Northampton, MA 01063
} } phone (413) 585-3765
} } fax (413) 585-3786
} } e-mail: mvolling-at-smith.edu
} }
} }
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From: bozzola-at-siu.edu
Date: Thu, 2 Sep 2010 08:30:36 -0500
Subject: [Microscopy] EM: where to buy 50 mesh coated grids

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

Paul, Phil and others who have generously shared your experiences
making Formvar coated grids:

Thanks for recalling your sometimes wild experiences making Formvar
coated grids. Paul, your description is priceless.

Everyone has inspired me to purchase some new Formvar powder, prepare
a 1% solution in ethylene dichloride (stirred, not shaken), and using
freshly cleaved mica or recently opened but not too clean slide
(possibly coated with a release substance), lowering the scribed slide
(within 5 minutes) into a water bath.

I shall report back regarding the success of this venture. In the
meantime, others are invited to send along any tips/tricks, etc. We
shall be looking forward to reading Paul's article on the procedure in
Microscopy Today...........

Seriously, though, I appreciate your suggestions.

JB

--
John J. Bozzola, Ph.D., Director of such films as Forever Formvar,
Great Expectations, Please Release Me
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


On Thu, Sep 2, 2010 at 8:04 AM, {oshel1pe-at-cmich.edu} wrote:
}
} John,
}
} Not a source of 50 mesh grids, but ... what slides are you using?
} We find that we have to use the really cheap
} P11011 generic chinese slides to get consistent
} release. These are bought in bulk for the Intro
} Bio course from VWR (I think - I'd have to ask
} the Intro Bio guy). Anything else is too good a
} quality to release well, even with the nose trick.
} The PacSci P11011 slides should work, but the
} cheapie Bellweather brand slides don't release.
}


==============================Original Headers==============================
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From: marcia.ventura-at-dq.fct.unl.pt
Date: Thu, 2 Sep 2010 09:49:46 -0500
Subject: [Microscopy] viaWWW: Sensitivity values for TEM-EDS

Contents Retrieved from Microscopy Listserver Archives
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Email: marcia.ventura-at-dq.fct.unl.pt
Name: Marcia Ventura

Organization: FCT-Universidade Nova de Lisboa

Title-Subject: [Filtered] Sensitivity values for TEM-EDS

Message: Dear Sirs,

Could you please tell me sensitivity values for the elements chlorine
and oxygen on the TEM-EDS method?

Thank you in advance.

Marcia Ventura

Login Host: 193.136.125.97
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From: colijn.1-at-osu.edu
Date: Thu, 2 Sep 2010 09:56:41 -0500
Subject: [Microscopy] Re: EM: where to buy 50 mesh coated grids

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

Hi Paul,

There is a lot of anecdotal evidence on hte best way to remove formvar
films from the casting substrate. I was shown the "nose grease" method
when I was starting out in the field. As I mentioned, mica seems to be
much more reliable than glass slides as a substrate. Also, adding
grease to the substrate runs counter to my practice of absolute
cleanliness on my samples.

Regarding formvar films... I generally use formvar only for my holey
(not holy) or lacy support films since formvar films are generally too
thick for my purposes. A chemistry colleague (note the anecdotal
evidence) told me that dichloroethane degrades with time forming
hydrochloric acid. This degradation is accelerated by light. The up
shot is that one should store the formvar/dichlorethane solution in a
dark bottle in a dark storage cabinet and make up fresh solution
periodically. Certainly my films were much stronger with the fresh
solution than with my old stuff.

I use the huffing technique (ala Arte Johnson) after casting the films
to create holes, rather than to serve as a separation layer. Since the
moisture in your breath is immiscible in the dichloroethane solution,
you get droplet formation and, after the water evaporates, holes. No
garlic, escargot or butter required!

Cheers,
Henk


At 9/2/2010 1:05 AM, paul_hazelton-at-umanitoba.ca wrote:
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} John
}
} Henk mentioned the nose grease thing on the list server a number of
} years ago. Tried it. May work for him, it was a total flop for me.
}
} We had trouble off and on for years. Super cleaned the slides, put
} grease on them, etc. One thing I noticed is how recently the slides
} were made seemed to have an effect. Invariably, we have trouble when
} the box of slides is old. The second they start to go milky, have
} whitish spots, etc, we cannot get the slides off. But slides from a new
} box, recently purchased, consistently will release the film. The other
} problem we've found is using slides that are super clean - acetone,
} ethanol, etc. But as Markus said, a slide fresh out of the box, wiped
} with a kimwipe works best (actually, I use a lab coat sleeve, but then
} I've been accused of being some form of reprobate anyway).
}
} The second thing is how long the formvar has been on the slide.
} Actually tried this silly little experiment just because I got curious
} as to whether I could make up a box of slides at one time, and use them
} over the next several months. Dipped slides and tried floating the
} formvar off after 10, 15, 30, 45, 60, 90, 120mins, 4, 8, 16, 24, and 48
} hours. Did 10 slides each.
}
} Any where from 10 to 30 minutes was great, after that slides started to
} give a little trouble. About 75% of slides released the formvar after 2
} hours. By 16 hours, almost no slides released the formvar. So making
} the film on slides in advance, and casting them into the water days or
} months later is just not going to work, at least in our hands.
}
} Breathing on the slides. Yep, it works. Just think back and do your
} Artie Johnson dirty old man bit from 'Laugh In' days. Actually, you
} don't have to breath that heavily. And if the family and techs don't
} mind, and you really like the escargots bourguignon, then go for it.
} And sop up the butter with that chunk of baguette. Make the process of
} making slides a celebratory excuse.
}
} Also, what are you making them in. We use ethylene dichloride. Have
} seen other solvents recommended, never tried them. Make it up 100ml at
} a time, pour it back into the coplin jar after each use - now everyone
} can scream about contamination, bad technique, etc - but it works. Be
} practical, help limit the amount of pollution you put into the
} environment, and stay out of the clutches of your Health and Safety
} Office. We dip about 12 slides, and immediately pour the formvar back
} into a bottle, seal with parafilm, and wait to use it later. I have
} used it for up to 2 years. Having said that, I also always float off
} the first film, put a single grid on it, and then check that grid in the
} microscope immediately to make sure there are no problems with the
} formvar solution before I make up 400-500 grids.
}
} We are still making up over 3,000 grids a year. Since the tech that was
} soooo good at making them became ill and passed away I have just made
} them myself. Too much trouble getting the other techs to get it right.
} I make up 1200 at a time, and make up a batch every time we get down
} to 200. Good mindless way to waste a Monday morning. Never had a
} problem with longevity - but then the grids never stay around more that
} 6 months at tops.
}
} Final point. Someone once asked me if I didn't find Formvar to be weak
} compared to collodion. A true gentleman and brilliant virologist -
} suspect the question was a "Hazelton, hmm, heard about him, how bright
} is he really" type of test. I made the observation then, as now, that
} all plastics are essentially the same. If your films are weak, just
} make a higher concentration solution next time. And if the lab you go
} into insists one plastic is better than another, go with the flow. Not
} worth the argument, and like politics and religion, you will never
} change a primal belief.
}
} And I've been so good the last several years, sticking to the
} gastroenteric molecular work, not polluting the band width, etc.....
} Next thing you know Phil's going to send me a note asking if I will put
} it together as a technique thing for Microscopy Today, and I still owe
} him something he asked me for several years ago, and cannot remember
} what it was for the life of me.
}
} Oh yeah, work avoidance, that's why I stopped polluting the band width....
}
} Paul

--
Hendrik O. Colijn
www.ceof.ohio-state.edu
OSU Campus Electron Optics Facility colijn.1-at-osu.edu
040 Fontana Labs (614) 292-0674 (V)
116 W. 19th Ave. (614) 292-7523 (F)
Columbus, OH 43210

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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From: PhillipsT-at-missouri.edu
Date: Thu, 2 Sep 2010 10:03:46 -0500
Subject: [Microscopy] dehydration

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

I have two questions about dehydration. I seem to remember a solvent that could be used after ethanol that reacted with any free water to generate acetone. Can someone identify the solvent and comment on its effectiveness? Secondly, can anyone comment on the use of n-butyl alcohol following ethanol as a dehydration step? Thanks. Tom


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/




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From: W.Muss-at-salk.at
Date: Thu, 2 Sep 2010 10:50:07 -0500
Subject: [Microscopy] AW: dehydration (+ Lit.Refs)

Contents Retrieved from Microscopy Listserver Archives
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Dear Prof. Philips,

just (hopefully!) not to be too long: you are addressing an interesting spec-prep-matter:

perhaps you meant "rapid dehydration from water to the organic phase "within seconds" (chemically this has been verified for small pieces of tissue, i. e. 1 mm3):

try: 2,2-Dimethoxypropane (2,2-DMP) acidified:

cf. (choices)

MULLER L.L. and JACKS T.J. 1975. Rapid chemical dehydration of samples for electronmicroscopic examinations. J. Histochem. Cytochem. 23: 107-110

POSTEK M. T. and TUCKER S. C. 1976. A new short chemical dehydration method for light microscopy preparations of plant material. Can. J. Bot. 54, 872-875

MASER D.M. and TRIMBLE III, J.J. 1977. Rapid chemical dehydration of biologic samples for scanning electron microscopy using 2,2-dimethoxypropane. J. Histochem. Cytochem. 25:247-251

LIN C.H., Falk R.H. and Stocking C.R. 1977. Rapid chemical Ddehydration of plant material for light and electron microscopy with 2,2-dimethoxypropane and 2,2-diethoxypropane. Am. J. Bot.64: 602-605

A. De RUITER, P. Van Banning and J.J. Willemse 1981
Brief technical note
Rapid histological results in aquaculture research by using the time-saving embedding procedure with 2,2-dimethoxypropane
Aquaculture, Volume 25, Issues 2-3, August 1981, Pages 293-297
[Abstract: Dehydration and clearing by 2,2-dimethoxypropane (DMP) is completed in 30 min. Tissues of eel and oysters, both of importance for aquaculture purposes, were used in a comparison of the DMP procedure and conventional dehydration-clearing. We found that the cutting quality and the histological appearance of the tissues were quite similar. The time-saving aspects are considered as an advantage of DMP for rapid results in aquaculture research]

KAESER W. 1989. Freeze substitution of plant tissues with a new medium containing dimethoxypropane. J. Microsc. 154: 273-278

MÖLLER W. and MÖLLER G. 1994. Chemical dehydration for rapid paraffin embedding. Biotech. & Histochem. 69: 289-290

HALBRITTER H. 1998: Preparing Living Pollen Material for SEM Using 2,2-DMP and Critical Point Drying;
Biotechnic & Histochemistry 73/#3, 137-143,

A PERNSTICH, H W Krenn, G Pass 2003. Preparation of serial sections of arthropods using 2,2-dimethoxypropane dehydration and epoxy resin embedding under vacuum
Biotechnic & Histochemistry, Jan 2003, Vol. 78, No. 1, Pages 5-9.


More:
cf. also ==} Google: {Lynn and "rapid dehydration" and 2,2-DMP} ca. 47 results

I have used this agent 2,2-DMP a long time ago for special cases where speed in processing was a serious question.
Elegant method, for introduction of the method do parallel spec.-preparations initillay for comparison. One should consider some changes in the preservation of usually expected ultrastructural morphology aspects after "classical" processing with EtOH ==} organic (like PO or Acetonitrile).
The last of at least 2 incubations with 2,2-DMP acidified should be followed by at least 2 "washes" in pure 2,2-DMP, the solvent mixes (initially) slowly to well with epoxide like Epon (but: with the "old, original Epon 812" in the late 80ies/starting 90ies I found it to be somehow tricky, mostly upset because of unusual / unexpected disturbances in the resin mixture (at least in the first mix 1:1==} so mix thoroughly). Other resin types I have not tested with that method.


n-Butyl-alcohol: sorry, no experience
more:
==} Google: { n-butyl alcohol and dehydration } to start with.



Best wishes and good luck


Wolfgang MUSS
EM-LAb,
Pathology SALK-LKH (Gen.Hosp)
SALZBURG-AUSTRIA





} -----Ursprüngliche Nachricht-----
} Von: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
} Gesendet: Donnerstag, 02. September 2010 17:07
} An: Muß Wolfgang
} Betreff: [Microscopy] dehydration
}
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}
} I have two questions about dehydration.
} I seem to remember a solvent that could be used after ethanol that
} reacted with any free water to generate acetone.
} Can someone identify the solvent and comment on its effectiveness?
}
} Secondly, can anyone comment on the use of n-butyl alcohol following } ethanol as a dehydration step?
}
} Thanks. Tom
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://www.biology.missouri.edu/faculty/phillips.html
} http://www.biotech.missouri.edu/mcc/
}
}
}
}
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From: isabel.nogueira-at-ist.utl.pt
Date: Thu, 2 Sep 2010 11:07:30 -0500
Subject: [Microscopy] viaWWW: Material for cement sample coating

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Email: isabel.nogueira-at-ist.utl.pt
Name: Isabel Dias Nogueira

Organization: ICEMS/IST

Title-Subject: [Filtered] Material for cement sample coating

Message: Dear microscopists:
I work in a lab with a FEG-SEM JEOL 7001F, and would like to ask for
advice for the purchase of a sputter coating system.
I need to observe and analyse (EDS) cement materials, being
particularly interested in the Si/Ca ratio and particle sizing. The
issue is what material would be best suited to coat the mounted and
polished samples: carbon, gold, platinum?... Carbon would not
interfere with any sample peaks, but I'm not sure whether its grain
size could interfere with small particle observation. Gold and
platinum peaks are rather close to the silicon peak, although once
again I don't know if they are close enough to cause any
deconvolution difficulties.
Does anyone have any input on this subject?

Thank very much in advance!

Isabel

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From: baskin-at-bio.umass.edu
Date: Thu, 2 Sep 2010 11:39:38 -0500
Subject: [Microscopy] Re: dehydration

Contents Retrieved from Microscopy Listserver Archives
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Tom,
I think you are refering to dimethoxy propane. Once the
compound is acidified, you can add it to organic solvens (eg actetone
or ethanol) and it reacts with water to make methanol and acetone
(one of each, I think). It is stoichiometric. So if you use 1% DMP
then you can remove 1% of water (roughly). I use it in freeze
substitution to remove a little extra water but I have not checked
rigorously to compare its use plus and minus, and in all fairness my
demands on the freeze sub process are modest. I think there are
papers (1980s?) where DMP was evaluated rigorously. I don't have them
to hand, sorry.
Hope this helps,
Tobias

}
} ----------------------------------------------------------------------------
}
} I have two questions about dehydration. I seem to remember a
} solvent that could be used after ethanol that reacted with any free
} water to generate acetone. Can someone identify the solvent and
} comment on its effectiveness? Secondly, can anyone comment on the
} use of n-butyl alcohol following ethanol as a dehydration step?
} Thanks. Tom
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://www.biology.missouri.edu/faculty/phillips.html
} http://www.biotech.missouri.edu/mcc/
}

--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Biology Department
/_ / __ /__ \ \ \__ 611 N. Pleasant St.
/ / / \ \ \ University of Massachusetts
/ / / \ \ \ Amherst, MA, 01003
/ / ___ / \ \__/ \ ____
www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

==============================Original Headers==============================
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From: oshel1pe-at-cmich.edu
Date: Thu, 2 Sep 2010 11:42:24 -0500
Subject: [Microscopy] Re: EM: where to buy 50 mesh coated grids

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

Paul,

You rang?
Well, since you already did both the initial contact and the
follow-up nag, I don't have to do anything, right? Wonderful,
wonderful, thank you for making this so easy. I'll just sit here with
my feet up and my glass of Elijah Craig, and peruse my copy of
"Illustrated Adventures of Dear Abbe and Uncle Rayleigh: The Rogue
Optician Strikes", while the articles pour in.
(Is that the mail I hear?)

Phil

} John
}
} Henk mentioned the nose grease thing on the list server a number of
} years ago. Tried it. May work for him, it was a total flop for me.
}
} We had trouble off and on for years. Super cleaned the slides, put
} grease on them, etc. One thing I noticed is how recently the slides
} were made seemed to have an effect. Invariably, we have trouble when
} the box of slides is old. The second they start to go milky, have
} whitish spots, etc, we cannot get the slides off. But slides from a new
} box, recently purchased, consistently will release the film. The other
} problem we've found is using slides that are super clean - acetone,
} ethanol, etc. But as Markus said, a slide fresh out of the box, wiped
} with a kimwipe works best (actually, I use a lab coat sleeve, but then
} I've been accused of being some form of reprobate anyway).
}
} The second thing is how long the formvar has been on the slide.
} Actually tried this silly little experiment just because I got curious
} as to whether I could make up a box of slides at one time, and use them
} over the next several months. Dipped slides and tried floating the
} formvar off after 10, 15, 30, 45, 60, 90, 120mins, 4, 8, 16, 24, and 48
} hours. Did 10 slides each.
}
} Any where from 10 to 30 minutes was great, after that slides started to
} give a little trouble. About 75% of slides released the formvar after 2
} hours. By 16 hours, almost no slides released the formvar. So making
} the film on slides in advance, and casting them into the water days or
} months later is just not going to work, at least in our hands.
}
} Breathing on the slides. Yep, it works. Just think back and do your
} Artie Johnson dirty old man bit from 'Laugh In' days. Actually, you
} don't have to breath that heavily. And if the family and techs don't
} mind, and you really like the escargots bourguignon, then go for it.
} And sop up the butter with that chunk of baguette. Make the process of
} making slides a celebratory excuse.
}
} Also, what are you making them in. We use ethylene dichloride. Have
} seen other solvents recommended, never tried them. Make it up 100ml at
} a time, pour it back into the coplin jar after each use - now everyone
} can scream about contamination, bad technique, etc - but it works. Be
} practical, help limit the amount of pollution you put into the
} environment, and stay out of the clutches of your Health and Safety
} Office. We dip about 12 slides, and immediately pour the formvar back
} into a bottle, seal with parafilm, and wait to use it later. I have
} used it for up to 2 years. Having said that, I also always float off
} the first film, put a single grid on it, and then check that grid in the
} microscope immediately to make sure there are no problems with the
} formvar solution before I make up 400-500 grids.
}
} We are still making up over 3,000 grids a year. Since the tech that was
} soooo good at making them became ill and passed away I have just made
} them myself. Too much trouble getting the other techs to get it right.
} I make up 1200 at a time, and make up a batch every time we get down
} to 200. Good mindless way to waste a Monday morning. Never had a
} problem with longevity - but then the grids never stay around more that
} 6 months at tops.
}
} Final point. Someone once asked me if I didn't find Formvar to be weak
} compared to collodion. A true gentleman and brilliant virologist -
} suspect the question was a "Hazelton, hmm, heard about him, how bright
} is he really" type of test. I made the observation then, as now, that
} all plastics are essentially the same. If your films are weak, just
} make a higher concentration solution next time. And if the lab you go
} into insists one plastic is better than another, go with the flow. Not
} worth the argument, and like politics and religion, you will never
} change a primal belief.
}
} And I've been so good the last several years, sticking to the
} gastroenteric molecular work, not polluting the band width, etc.....
} Next thing you know Phil's going to send me a note asking if I will put
} it together as a technique thing for Microscopy Today, and I still owe
} him something he asked me for several years ago, and cannot remember
} what it was for the life of me.
}
} Oh yeah, work avoidance, that's why I stopped polluting the band width....
}
} Paul
} --
} Paul R. Hazelton, PhD
} Viral Gastroenteritis Study Group
} University of Manitoba
} Department of Medical Microbiology
} 511 Basic Medical Sciences Building
} 745 Bannatyne Avenue
} Winnipeg, Manitoba, Canada, R3E 3J7
} e-mail: paul_hazelton-at-umanitoba.ca
} paulhazelton-at-mts.net
} Phone: 204-789-3313 (w);
} 204-489-6924 (h)
} Cell: 204-781-6982
} Fax: 204-789-3926

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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From: FMonson-at-wcupa.edu
Date: Thu, 2 Sep 2010 12:52:10 -0500
Subject: [Microscopy] EM: where to buy 50 mesh coated grids

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

Tom,

URL: http://www.informaworld.com/smpp/content~db=all~content=a787850862

Chemical Dehydration for Rapid Paraffin Embedding
Authors: W. Mllera; G. Mllera
Affiliation: Department of Anatomy and Cytobiology, Giessen, Germany
DOI: 10.3109/10520299409106304
Publication Frequency: 6 issues per year
Published in: Biotechnic and Histochemistry, Volume 69, Issue 5 September 1994 , pages 289 - 290

Now Paul,

If you look at your address and the number of phone numbers, you have a case for BWP (Band Width Pollution), however, everything else is useful - and quite mystical, which is just the ticket for those of us who have 'Monkish' laboratory 'best' practices - at least when WE are working at the bench.

I once used the placement of grids on a black Formvar film on water in a black Bakelite developer dish to instruct a class of undergraduates in a typical mystery of microtechnique. Out of 12 students who watched me demonstrate, we made it thru two before the third tangled the forceps in the film and proved its existence.

Reminds me of the time I demonstrated the distinction between dis-section and di-section by using a 30" machete to bisect a 24" shark along the long axis with the shark smiling at the class from the front lab bench - at least until its halves tilted to each side. I ended my classroom teaching long before students and instructors started describing the NUCULI of cells and NUCULAR ENERGY. I have not yet decided how to attack such anti-scholarly habits when they appear in my basement microscope lab. I have started to introduce myself to new microscopy trainees by asking them to enumerate all of the digital cameras they have brought with them to my lab.

Thanks for the opportunity to REALLY increase BWP,

Fred

Frederick C. Monson, PhD
Technical Director, CMIRT
West Chester University
West Chester, PA, 19383
610-738-0437
New Web Site:           http://cmirt.wcupa.edu/index.html

New Scheduler:          http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl

Reads of the Month:
1. "The Liver as a Lymphoid Organ," Ian Nicholas Crispe, Annual Review of Immunology, Apr 2009, Vol. 27: 147-163. (Access Depends on subscription.)
2. "A Personal Journey of Discovery: Developing Technology and Changing Biology," Lee Hood, Annual Review of Analytic Chemistry, Vol. 1: 1-43.


-----Original Message-----
X-from: paul_hazelton-at-umanitoba.ca [mailto:paul_hazelton-at-umanitoba.ca]
Sent: Thursday, September 02, 2010 1:13 AM
To: Monson, Frederick

John

Henk mentioned the nose grease thing on the list server a number of
years ago. Tried it. May work for him, it was a total flop for me.

We had trouble off and on for years. Super cleaned the slides, put
grease on them, etc. One thing I noticed is how recently the slides
were made seemed to have an effect. Invariably, we have trouble when
the box of slides is old. The second they start to go milky, have
whitish spots, etc, we cannot get the slides off. But slides from a new
box, recently purchased, consistently will release the film. The other
problem we've found is using slides that are super clean - acetone,
ethanol, etc. But as Markus said, a slide fresh out of the box, wiped
with a kimwipe works best (actually, I use a lab coat sleeve, but then
I've been accused of being some form of reprobate anyway).

The second thing is how long the formvar has been on the slide.
Actually tried this silly little experiment just because I got curious
as to whether I could make up a box of slides at one time, and use them
over the next several months. Dipped slides and tried floating the
formvar off after 10, 15, 30, 45, 60, 90, 120mins, 4, 8, 16, 24, and 48
hours. Did 10 slides each.

Any where from 10 to 30 minutes was great, after that slides started to
give a little trouble. About 75% of slides released the formvar after 2
hours. By 16 hours, almost no slides released the formvar. So making
the film on slides in advance, and casting them into the water days or
months later is just not going to work, at least in our hands.

Breathing on the slides. Yep, it works. Just think back and do your
Artie Johnson dirty old man bit from 'Laugh In' days. Actually, you
don't have to breath that heavily. And if the family and techs don't
mind, and you really like the escargots bourguignon, then go for it.
And sop up the butter with that chunk of baguette. Make the process of
making slides a celebratory excuse.

Also, what are you making them in. We use ethylene dichloride. Have
seen other solvents recommended, never tried them. Make it up 100ml at
a time, pour it back into the coplin jar after each use - now everyone
can scream about contamination, bad technique, etc - but it works. Be
practical, help limit the amount of pollution you put into the
environment, and stay out of the clutches of your Health and Safety
Office. We dip about 12 slides, and immediately pour the formvar back
into a bottle, seal with parafilm, and wait to use it later. I have
used it for up to 2 years. Having said that, I also always float off
the first film, put a single grid on it, and then check that grid in the
microscope immediately to make sure there are no problems with the
formvar solution before I make up 400-500 grids.

We are still making up over 3,000 grids a year. Since the tech that was
soooo good at making them became ill and passed away I have just made
them myself. Too much trouble getting the other techs to get it right.
I make up 1200 at a time, and make up a batch every time we get down
to 200. Good mindless way to waste a Monday morning. Never had a
problem with longevity - but then the grids never stay around more that
6 months at tops.

Final point. Someone once asked me if I didn't find Formvar to be weak
compared to collodion. A true gentleman and brilliant virologist -
suspect the question was a "Hazelton, hmm, heard about him, how bright
is he really" type of test. I made the observation then, as now, that
all plastics are essentially the same. If your films are weak, just
make a higher concentration solution next time. And if the lab you go
into insists one plastic is better than another, go with the flow. Not
worth the argument, and like politics and religion, you will never
change a primal belief.

And I've been so good the last several years, sticking to the
gastroenteric molecular work, not polluting the band width, etc.....
Next thing you know Phil's going to send me a note asking if I will put
it together as a technique thing for Microscopy Today, and I still owe
him something he asked me for several years ago, and cannot remember
what it was for the life of me.

Oh yeah, work avoidance, that's why I stopped polluting the band width....

Paul
--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 Bannatyne Avenue
Winnipeg, Manitoba, Canada, R3E 3J7
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926


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From: FMonson-at-wcupa.edu
Date: Thu, 2 Sep 2010 13:00:44 -0500
Subject: [Microscopy] viaWWW: Material for cement sample coating

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

Isabel,

An evaporated Carbon coating should be the coating of choice, and if you must use a metal, use gold alone. I have Oxford INCA and can, for each 'Sample' specify a single coating (thickness) whose spectral effect will then be subtracted from the calculations. With a vacuum evaporator, you can also coat with gold or platinum (or other metals).
I would recommend a sputter coater with a thickness monitor if you are going to sputter your coating. For carbon, we really like our evaporative coater. I have no experience with sputtering carbon, so I cannot offer a comparison.

Cheers,

Fred Monson

Frederick C. Monson, PhD
Technical Director, CMIRT
West Chester University
West Chester, PA, 19383
610-738-0437
New Web Site:           http://cmirt.wcupa.edu/index.html

New Scheduler:          http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl

Reads of the Month:
1. "The Liver as a Lymphoid Organ," Ian Nicholas Crispe, Annual Review of Immunology, Apr 2009, Vol. 27: 147-163. (Access Depends on subscription.)
2. "A Personal Journey of Discovery: Developing Technology and Changing Biology," Lee Hood, Annual Review of Analytic Chemistry, Vol. 1: 1-43.


-----Original Message-----
X-from: isabel.nogueira-at-ist.utl.pt [mailto:isabel.nogueira-at-ist.utl.pt]
Sent: Thursday, September 02, 2010 12:16 PM
To: Monson, Frederick

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Email: isabel.nogueira-at-ist.utl.pt
Name: Isabel Dias Nogueira

Organization: ICEMS/IST

Title-Subject: [Filtered] Material for cement sample coating

Message: Dear microscopists:
I work in a lab with a FEG-SEM JEOL 7001F, and would like to ask for
advice for the purchase of a sputter coating system.
I need to observe and analyse (EDS) cement materials, being
particularly interested in the Si/Ca ratio and particle sizing. The
issue is what material would be best suited to coat the mounted and
polished samples: carbon, gold, platinum?... Carbon would not
interfere with any sample peaks, but I'm not sure whether its grain
size could interfere with small particle observation. Gold and
platinum peaks are rather close to the silicon peak, although once
again I don't know if they are close enough to cause any
deconvolution difficulties.
Does anyone have any input on this subject?

Thank very much in advance!

Isabel

Login Host: 193.136.142.7
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From: maryflet-at-interchange.ubc.ca
Date: Thu, 2 Sep 2010 13:25:57 -0500
Subject: [Microscopy] viaWWW: Material for cement sample coating

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Dear Isabel,
I find that 60% gold-40% palladium coating is the best for both
medium-to-high resolution imaging and EDS analysis, but if you really want
high resolution on tiny particles, the Pt coating is probably better. The
gold-palladium peaks are tiny and do not interfere with Si or Ca. The carbon
coating grain size is not an issue, since you do not see the carbon coating,
but I find it is difficult to get good high-resolution imaging with a carbon
coated sample, since there is not much signal. When I want high-resolution
imaging and EDS on the same sample, I make two samples and use one for
imaging and one for analysis. Variable-pressure mode and no coating also
works for EDS, but is not as precise for analyzing one area without getting
stray x-rays from another, due to the beam scattering by the gas.
I also find that concrete samples out-gas badly and you are best to stick to
small samples if you want to work at high vacuum.
IMHO
Regards,

Mary Fletcher
Electron Microscopist
Materials Eng. UBC
#309 - 6350 Stores Road
Vancouver, B.C. V6T 1Z4
Canada
Tel: 604-822-5648
Fax: 604-822-3619
email: maryflet-at-interchange.ubc.ca


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Email: isabel.nogueira-at-ist.utl.pt
Name: Isabel Dias Nogueira

Organization: ICEMS/IST

Title-Subject: [Filtered] Material for cement sample coating

Message: Dear microscopists:
I work in a lab with a FEG-SEM JEOL 7001F, and would like to ask for
advice for the purchase of a sputter coating system.
I need to observe and analyse (EDS) cement materials, being
particularly interested in the Si/Ca ratio and particle sizing. The
issue is what material would be best suited to coat the mounted and
polished samples: carbon, gold, platinum?... Carbon would not
interfere with any sample peaks, but I'm not sure whether its grain
size could interfere with small particle observation. Gold and
platinum peaks are rather close to the silicon peak, although once
again I don't know if they are close enough to cause any
deconvolution difficulties.
Does anyone have any input on this subject?

Thank very much in advance!

Isabel

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From: tina-at-pbrc.hawaii.edu
Date: Thu, 2 Sep 2010 13:31:16 -0500
Subject: [Microscopy] Re: EM: where to buy 50 mesh coated grids

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

Everyone's got their tricks. Mine include the following:

1) I buy special-order 8% Formvar from Ladd (long story about how this
came about), then I dilute to about 0.8 - 1% to use (or whatever the math
in #2, below, works out to). This seems to be our magic solution. I never
make from powder any more.

2) I do this in a Dip-Miser beaker so that I only have to make about 11 ml
(1ml 8% Formvar plus about 10ml ethylene dichloride).

3) I have the Dip-Miser propped up in a beaker, nestled in dessicant in
the bottom of a peanut butter jar, the metal top of which has a hinged
trap door cut into it, beaker surrounded by gauze well moistened with
ethylene dichloride. The top is held down by a dive weight when I'm not
dipping - I actually think this is the crucial component ;-)

4) Our "magic slides" change every couple of years. The current box is
three-year-old plain Corning slides that do not seem to get frosty as they
age. I wipe not too well with Kimwipe. Have been know to use nose grease,
Ivory hand soap, whatever feels right that day.

5) Because of the humidity here, I dip the slide, let the Formvar run off
it a bit back into the beaker in the ethylene dichloride atmosphere
(amount of time helps determine film thickness) with the trap door as shut
around my hand as I can get it, then I quickly put it to dry on filter
paper that is on top of dessicant in another jar.

6) Strip and float and add grids as usual. I pick up from the top with
Parafilm which, once I tried it, makes me very, very happy.

7) In my hands, really old coated grids (} 2 years, preferably about 6
years) are happily hydrophilic. Otherwise I fire up the Denton carbon
evaporator with a bad leak which just happens to be the perfect leak rate
for glow discharging the grids. This is my excuse for not repairing it.

Aloha,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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From: kaszas.1-at-osu.edu
Date: Thu, 2 Sep 2010 14:23:01 -0500
Subject: [Microscopy] viaWWW:microtome sections thickness problem

Contents Retrieved from Microscopy Listserver Archives
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None of the elements should cause a problem with determining the intensity ratio between the Ca and Si peaks. Current deconvolution software should easily handle any minor overlap that might occur. Pt and Au could cause some problem with P or S. Pd could cause problems with Cl. C should cause no problems.

Your SEM has a field emission gun so it should have no problem seeing the texture in the gold layer if you get into the tens of thousands of magnifications. I understand Au-Pd alloys and Pt give finer structures. I have not gone looking for structure in C layers. I understand high vacuum evaporation gives better films than does the flash evaporation in a sputter coater.

I also understand that you will be using polished samples. Do be careful in polishing. I presume you will not be using water since it would react with the cement.

Polished sample will allow for the best EDS analyses and will simplify image analyses. EDS on rough particles is very difficult. You would see the Si/Ca ratio bounce all over depending on the geometry. The geometry will also affect the brightness of the image and make particle detection and sizing more difficult.

I presume you will be using BSE imaging. The particles will readily stand out from the embedding media. C would present the least interference with the available contrast. The metals might reduce (mask) the contrast a little but it should be a uniform effect.

Written by one who has looked at a few cement samples in my years,
Warren

-----Original Message-----
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Email: kaszas.1-at-osu.edu
Name: Andrea Kaszas

Organization: Ohio State University/OARDC

Title-Subject: [Filtered] sections thickness problem

Message: Dear Microscopist,

I have problems with the sectioning thickness nowadays. We have two
Leica microtomes. When we are sectioning thick or thin sections one
of the section is thicker and the second one is much thinner after
this a thicker section come again and a thinner after that and so on.
Have you ever been this kind of problem before? What can cause that problem?

Any suggestion would be greatly appreciated.

Thank you for your help in advance.

Andrea Kaszas

microscope technician

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From: DusevichV-at-umkc.edu
Date: Thu, 2 Sep 2010 14:23:34 -0500
Subject: [Microscopy] RE: viaWWW: Material for cement sample coating

Contents Retrieved from Microscopy Listserver Archives
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Isabel,
To get best results from your microscope you need both metal and carbon
coating equipment. You can buy sputter coater with carbon coating device
as an option (to save money). To be on a minimalist side, I think you
need carbon coater. For particle measurements you can get excellent
results from polished sections with BSE in LV mode (no coating), but EDS
in LV mode, especially for small particles, is troublesome. Carbon
coating is superior for quantifying EDS results, like Si/Ca ratio
calculations. You can expect pretty good imaging with carbon coating up
to magnifications of about 10k. For higher magnifications, if you do
not want to spend money on high-resolution spatter coaters, you can by
conventional coater with Au-Pd target (and conventional coater is much
handier). It works fine for me up to magnifications 50k. Both Au and Pd
peaks are far enough from you target elements, so that interference of
small peaks from coating is negligible. But if you are going to use
metals for coating, for quantification purposes you should use as thin
coating as possible and coat your standards as well.

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

} -----Original Message-----
} From: isabel.nogueira-at-ist.utl.pt [mailto:isabel.nogueira-at-ist.utl.pt]
} Sent: Thursday, September 02, 2010 11:08 AM
} To: Dusevich, Vladimir
} Subject: [Microscopy] viaWWW: Material for cement sample coating
}
}
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} Email: isabel.nogueira-at-ist.utl.pt
} Name: Isabel Dias Nogueira
}
} Organization: ICEMS/IST
}
} Title-Subject: [Filtered] Material for cement sample coating
}
} Message: Dear microscopists:
} I work in a lab with a FEG-SEM JEOL 7001F, and would like to ask for
} advice for the purchase of a sputter coating system.
} I need to observe and analyse (EDS) cement materials, being
} particularly interested in the Si/Ca ratio and particle sizing. The
} issue is what material would be best suited to coat the mounted and
} polished samples: carbon, gold, platinum?... Carbon would not
} interfere with any sample peaks, but I'm not sure whether its grain
} size could interfere with small particle observation. Gold and
} platinum peaks are rather close to the silicon peak, although once
} again I don't know if they are close enough to cause any
} deconvolution difficulties.
} Does anyone have any input on this subject?
}
} Thank very much in advance!
}
} Isabel
}
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} 8, 22 -- Subject: viaWWW: Material for cement sample coating
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From: tarawdan-at-ncsu.edu
Date: Thu, 2 Sep 2010 15:42:26 -0500
Subject: [Microscopy] Electron Microscopy Lab Manager Position

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

Electron Microscopy Laboratory Manager
Department of Materials Science and Engineering
NC State University, Raleigh, North Carolina

Permanent - Full-time   Time-Limited Position. One year w/ possible
extension  Continuation of position for a second year is subject to the
availability of funds.  
Essential Job Duties:
Manages certification, access and invoicing of Facility users. Provide
and/or oversee the care and preventive maintenance of the Facility's
instrumentation and computer hardware/software. Maintain environmental and
radiation safety compliance, as well as contracts and grants service center
compliance per OMB A27.
Train Facility users in the safe and proper use of the laboratories and
instruments. Provide educational support to the department's undergraduate
and graduate materials science and engineering courses dependent and/or
based on the Facility's analytical instrumentation.
Minimum Qualifications:
MS in Materials Science, Physics or related field with a minimum of three
years experience operating a TEM and performing TEM specimen preparation.
Two years experience in the care and preventive maintenance of electron
microscopes and specimen preparation instruments.  
Required Skills:
Able to align, operate and calibrate TEM instruments and prepare TEM
specimens is essential. Know how to acquire images using HRTEM, CBED,
BF/DF/WBDF and STEM techniques is essential. Must be knowledgeable in the
setup and use of personal computer hardware and software related to TEM
instruments and lab. Must be able to perform preventative maintenance on TEM
instruments and specimen preparation equipment (filament exchange, chillers,
vacuum pumps, diagnose vacuum problems, etc.). Possess a very good command
of speaking and writing the English language as good presentation skills are
essential.  
Analytical skills and abilities in characterization of condensed matter
materials using TEM, STEM, CBED, XEDS, EELS and SEM is a plus.
Anticipated Hiring Range: $50,000-$70,000  

To Apply:
Submit application on line at https://jobs.ncsu.edu/
Position Number: 101303  
Application Materials Required: Resume/CV, Cover Letter  

AA/EEO. In addition, NC State welcomes all persons without regard to sexual
orientation.



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From: wtivol-at-verizon.net
Date: Thu, 2 Sep 2010 15:50:27 -0500
Subject: [Microscopy] Re: EM: where to buy 50 mesh coated grids

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

Dear John,
An additional trick is to use hot water. When I was using Apiazon on
the slides, I also prepared a solution of Alconox, 1 g/l and put it in
the 70 deg oven. I used this in a staining dish when it came time to
float the formvar.
Yours,
Bill

On Sep 2, 2010, at 6:38 AM, bozzola-at-siu.edu wrote:

}
} Paul, Phil and others who have generously shared your experiences
} making Formvar coated grids:
}
} Thanks for recalling your sometimes wild experiences making Formvar
} coated grids. Paul, your description is priceless.
}
} Everyone has inspired me to purchase some new Formvar powder, prepare
} a 1% solution in ethylene dichloride (stirred, not shaken), and using
} freshly cleaved mica or recently opened but not too clean slide
} (possibly coated with a release substance), lowering the scribed slide
} (within 5 minutes) into a water bath.
}
} I shall report back regarding the success of this venture. In the
} meantime, others are invited to send along any tips/tricks, etc. We
} shall be looking forward to reading Paul's article on the procedure in
} Microscopy Today...........
}
} Seriously, though, I appreciate your suggestions.
}
} JB
}
} --
} John J. Bozzola, Ph.D., Director of such films as Forever Formvar,
} Great Expectations, Please Release Me
} I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
} Southern Illinois University
} 750 Communications Drive
} Carbondale, IL 62901
} Phone: 618-453-3730
}
}
} On Thu, Sep 2, 2010 at 8:04 AM, {oshel1pe-at-cmich.edu} wrote:
} }
} } John,
} }
} } Not a source of 50 mesh grids, but ... what slides are you using?
} } We find that we have to use the really cheap
} } P11011 generic chinese slides to get consistent
} } release. These are bought in bulk for the Intro
} } Bio course from VWR (I think - I'd have to ask
} } the Intro Bio guy). Anything else is too good a
} } quality to release well, even with the nose trick.
} } The PacSci P11011 slides should work, but the
} } cheapie Bellweather brand slides don't release.
} }


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From: bozzola-at-siu.edu
Date: Thu, 2 Sep 2010 16:02:04 -0500
Subject: [Microscopy] Re: viaWWW:microtome sections thickness problem

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

There are several possible causes.

One possibility is that the knife is not sharp enough to cut at the
setting you have selected, so it will skip a section or cut an overly
thick one followed by no section or a very thin one. Try increasing
the thickness setting to see if you get more consistent sections.
Sometimes, you just have to cut thicker sections (especially if you
have a soft embedding plastic).

Check the clearance angle of the knife holder. Sometimes, it gets
reset by other users, so the specimen rubs along the back of the
knife, compressing the block face. Then, when the block face starts
re-expanding, you get a thick section, followed by too thin of a
section.

If the block is not secured in the chuck, and the chuck in the arm,
then this will happen.

Excessive temperature fluctuations might cause this (though, it is
very unlikely).

A slipping belt in the stepper motor that advances the microtome might
cause this. I believe we currently have this problem with one of our
Leica UCT's and a Reichert Ultracut E. We are diagnosing it now.

Lastly, the most unpleasant possibility would be a problem with the
control electronics.

Let us know when you determine the cause. This will be helpful to others.

JB

On Thu, Sep 2, 2010 at 2:24 PM, {kaszas.1-at-osu.edu} wrote:
}
}
}
} ----------------------------------------------------------------------------
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} Email: kaszas.1-at-osu.edu
} Name: Andrea Kaszas
}
} Organization: Ohio State University/OARDC
}
} Title-Subject: [Filtered] sections thickness problem
}
} Message: Dear Microscopist,
}
} I have problems with the sectioning thickness nowadays. We have two
} Leica microtomes. When we are sectioning thick or thin sections one
} of the section is thicker and the second one is much thinner after
} this a thicker section come again and a thinner after that and so on.
} Have you ever been this kind of problem before? What can cause that problem?
}
} Any suggestion would be greatly appreciated.
}
} Thank you for your help in advance.
}
} Andrea Kaszas
}
} microscope technician
}
}  Login Host: 164.107.85.175
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} ==============================Original Headers==============================
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} 11, 22 -- Subject: viaWWW:microtome sections thickness problem
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}



--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


==============================Original Headers==============================
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From: greggps-at-umich.edu
Date: Thu, 2 Sep 2010 16:17:01 -0500
Subject: [Microscopy] Troubleshooting: TEM gun-Y drift. Need help with diagnosis.

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

Thanks to all who have responded. I really appreciate it.

I'd like to add a few details and see if they clarify things at all:

- I've changed out filaments, and the problem doesn't seem to be changed.
- The back and forth dancing of the beam was only present one or two days over the past 4-5 months, so that might have been charging.
- Mostly, the beam just occasionally moves while the scope is not actively being used.
- The physical Gun settings change in the display parameters.
- At one point, the Gun-Tilt-Y knob was actually moving the beam instead of intensifying the illumination. As of this morning, this had temporarily corrected itself. This was the detail that drove our engineer crazy, and leads us to believe it's an electronic issue.
- There doesn't seem to be any unusual beam behavior when we change the emission or the KV.
- Possibly unrelated: Our optimal Gun-Tilt settings are close to their extreme limit and have been so since this EM unit was turned off for several months, then moved to a new location and reactivated.

Do any two or more of these descriptions bring up any new theories?

Regards,
~Gregg

Gregg Sobocinski
Microscope Imaging Specialist
University of Michigan, MCDB Dept.
Ann Arbor, Michigan
USA

-----Original Message-----
X-from: greggps-at-umich.edu [mailto:greggps-at-umich.edu]
Sent: Wednesday, September 01, 2010 2:57 PM
To: Sobocinski, Gregg

Our Philips CM100 BioTwin TEM has developed an electronic defect, which causes the gun Y-axis setting to drift. Some days the beam is relatively stable, some days you can see the beam 'jump' back and forth up to a centimeter's distance on the viewing stage (moving once every second), and some days we'll turn on the filament to find the beam completely missing, requiring a reasonable amount of alignment (not just the "Y" direction.)

Our service provider has not seen this previously, and has moved around and tested the circuit boards with no effect on the problem. His last two guesses are expensive (Probably $4,000-$7,000, and we've already put quite a bit into troubleshooting and power supplies this year), so I wanted to know if anyone else has encountered this phenomenon, and how you fixed it.

If we could confidently choose which item is defective, I can probably budget for it, but those holding the purse strings are a little nervous right now.

Your input is greatly appreciated. All suggestions or guesses are welcome.  
~Gregg

Gregg Sobocinski
Microscope Imaging Specialist
University of Michigan, MCDB Dept.
Ann Arbor, Michigan
USA




==============================Original Headers==============================
8, 29 -- From greggps-at-umich.edu Wed Sep 1 13:49:28 2010
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8, 29 -- Date: Wed, 1 Sep 2010 14:49:23 -0400
8, 29 -- Subject: Troubleshooting: TEM gun-Y drift. Need help with diagnosis.
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From: y.han.1-at-bham.ac.uk
Date: Thu, 2 Sep 2010 16:18:40 -0500
Subject: [Microscopy] viaWWW: STEM over TEM

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Email: y.han.1-at-bham.ac.uk
Name: Yisong Han

Organization: University of Birmingham

Title-Subject: [Filtered] STEM over TEM

Message: Hi everyone,

Just a quick question. Is STEM more likely to achieve atomic
resolution over TEM on thick samples? Do you know any papers or
evidence about this? Thanks very much indeed in advance.

Yisong

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From: cvierret-at-mst.edu
Date: Thu, 2 Sep 2010 16:19:15 -0500
Subject: [Microscopy] viaWWW: GW Electronics

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Email: cvierret-at-mst.edu
Name: Clarissa Wisner

Organization: Missouri Universityof Science and Technology

Title-Subject: [Filtered] GW Electronics

Message: Greetings,

This is a two part query:

1. Does anybody have a current contact for the company GW
Electronics out of Georgia?

2. Does anybody know of a company in the US that can work on GW chamberscopes?

Our chamberscope has a vacuum leak and we would like to have it
repaired versus replacing it.

Thanks in advance for your replies

Clarissa

Login Host: 131.151.110.200
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From: wtivol-at-verizon.net
Date: Thu, 2 Sep 2010 16:19:58 -0500
Subject: [Microscopy] Re: viaWWW: Sensitivity values for TEM-EDS

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On Sep 2, 2010, at 8:01 AM, marcia.ventura-at-dq.fct.unl.pt wrote:

}
} Email: marcia.ventura-at-dq.fct.unl.pt
} Name: Marcia Ventura
}
} Organization: FCT-Universidade Nova de Lisboa
}
} Title-Subject: [Filtered] Sensitivity values for TEM-EDS
}
} Message: Dear Sirs,
}
} Could you please tell me sensitivity values for the elements chlorine
} and oxygen on the TEM-EDS method?
}
} Thank you in advance.
}
} Marcia Ventura
}
Dear Marcia,
Since Cl and O are both volatile, the sensitivities will depend on
many variables, such as the chemical forms (chlorides vs organo-
chlorine, etc.) the HT, the irradiation rate and total beam dose.
Some of these can be compensated for by taking measurements at several
doses and extrapolating to zero dose. It will also be useful to have
a standard with the same chemistry and about the same matrix
composition.
Yours,
Bill


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From: gary-at-gaugler.com
Date: Thu, 2 Sep 2010 16:42:57 -0500
Subject: [Microscopy] Re: [SPAM] viaWWW: GW Electronics

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

GW was sold to KE Development in the UK. Don't know
of US-based service.

Try "Gordon England" {gengland-at-kedev.com}

gary g.


At 02:21 PM 9/2/2010, you wrote:



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From: paul_hazelton-at-umanitoba.ca
Date: Fri, 3 Sep 2010 00:48:00 -0500
Subject: [Microscopy] Re: EM: where to buy 50 mesh coated grids

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

GW was sold to KE Development in the UK. Don't know
of US-based service.

Try "Gordon England" {gengland-at-kedev.com}

gary g.


At 02:21 PM 9/2/2010, you wrote:



} ----------------------------------------------------------------------------
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==============================Original Headers==============================
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From a1aaa1azzzz1zaaaaa-at-melarti.com Thu Sep 2 16:47:42 2010
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Received: from [41.199.27.100] by mail.global.frontbridge.com; Thu, 2 Sep 2010 23:44:04 +0200

Clarissa,
KE Developments bought them about 7 years ago. Try
http://www.kedev.co.uk/
or
Graham Wardall gwardall-at-kedev.com .

If the only problem is a vacuum leak, I might be able to help out.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
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Email: cvierret-at-mst.edu
Name: Clarissa Wisner

Organization: Missouri Universityof Science and Technology

Title-Subject: [Filtered] GW Electronics

Message: Greetings,

This is a two part query:

1. Does anybody have a current contact for the company GW
Electronics out of Georgia?

2. Does anybody know of a company in the US that can work on GW
chamberscopes?

Our chamberscope has a vacuum leak and we would like to have it
repaired versus replacing it.

Thanks in advance for your replies

Clarissa

Login Host: 131.151.110.200
---------------------------------------------------------------------------

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==============================Original Headers==============================
25, 28 -- From kenconverse-at-qualityimages.biz Thu Sep 2 17:39:51 2010
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Reply-To: {richowen2010-at-gmail.com}

Ed - et al

I am a true recidivist, if that is the correct word or concept. not
sure. My intellectual EM parent must hate me. He taught me so well,
where did I go wrong.

Buy the Ethylene dichloride from VWR. Could buy it anywhere. Throw in
a 4A molecular sieve, again, from VWR. Not using the ethylene
dichloride to dehydrate tissue for embedding, so there really are no
concerns of microscopic particles of the molecular sieve getting into
the tissue and wrecking the diamond knife.

I had trouble with getting dry ethylene dichloride for years. Would get
a new 500ml bottle, use it once, and even with molecular sieves it would
be bad the next time I opened it. Water in the solvent, leading to holy
grids (not holey, but holy as in holy -at-##%-at-!!!). All sorts of holes.
Then one summer day about 15 years ago I took a brand new bottle out of
the explosion safe fridge to make fresh formvar. In the summer the
humidity tends to be high in Manitoba, not as high as Tina has to fight
with, but high never the less, I came back 15 minutes later, after
weighing everything out, getting the volumentric flask ready, etc, and
picked up the bottle. It almost slipped out of my hand - coated with
condensation. In a brief and rare flirtation with brilliance I realized
that the problem of wet solvent was the result of proper storage -
keeping the solvent cold. When it came out of the Fridge moisture in
the air condensed in and out of the bottle, leading to wet ethylene
dichloride, and an excellent medium for making holey (holy) grids. Not
sure if there have been any such episodes of understanding since.

Today I am using a 500ml bottle that I bought 4 years ago. Keep it in
the Flammable Storage cabinet at room temperature. Never have a
problem. Probably will replace it next year since I do get worried
about it being too old.

As far as the breathing, it makes sense that if you immediately breath
on the film when you take the slide out of the formvar you should get
some holes in the film. We wait until the film is dry - usually 10
minutes, but I suspect even 5 minutes would work.

Have heard of using parafilm to pick up the grids, never tried it. We
pick up on a torn piece of paper towel. Let it get wet completely by
wicking, and the film and grids stay down. If we don't let the towel
get sufficiently wet, and the area of the film is not completely
bordered by wet towel, then we find invariably that the film will not
hold on the paper towel. Since we put 10-12 rows of 6 grids on a film,
I really do not like having the film and grids slide off the paper towel
- just because we tried to save 10-15 seconds.

In his post, Henk quite rightly points out something that I have also
used as an explanation for why the new slides work better than old ones.
It is my belief that the cheap surfactant or left over detergent on
the slides from time of manufacture makes the removal process work.
Over time this surfactant or detergent residue deteriorates, and is
visualized as the milky white material on the slides. Of course, I have
no hard data, so this hypothesis cannot be proven. Therefore, out of
the awe and respect that should be shown to sauride (saurine? is that
the right word or did I just make one up, oh well, dinosaur) electron
microscopists, it must be given status of dogma - not?

As far as degradation products, the MSDS sheet I have on file says it
degrades to Hydrogen chloride. Perhaps not HCl, but quite a nasty
little compound on it's own. Yep, keep it in brown glass.

And on that note, having polluted the band width enough for today, I
hear a glass (bottle?) of really nice montepulciano calling my name.
Wonder where those escargot are....

paul

--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 Bannatyne Avenue
Winnipeg, Manitoba, Canada, R3E 3J7
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926


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From: ehaller-at-health.usf.edu
Date: Fri, 3 Sep 2010 07:41:43 -0500
Subject: [Microscopy] viaWWW:microtome sections thickness problem

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

I did some google research as this DMP interested me.
I found both advantages and disadvantages.
I thought DMP may produce less extraction than ethanol but it does not seem to
be the case.

http://www.ncbi.nlm.nih.gov/pubmed/10190257
http://www.springerlink.com/content/m214012535m50403/
http://www.jhc.org/cgi/reprint/25/4/247.pdf

Regards,
Stephane


----- Original Message ----
X-from: "FMonson-at-wcupa.edu" {FMonson-at-wcupa.edu}
To: nizets2-at-yahoo.com
Sent: Thu, September 2, 2010 6:59:46 PM

Tom,

URL:  http://www.informaworld.com/smpp/content~db=all~content=a787850862

Chemical Dehydration for Rapid Paraffin Embedding
Authors: W. Mllera; G. Mllera
Affiliation: Department of Anatomy and Cytobiology, Giessen, Germany
DOI: 10.3109/10520299409106304
Publication Frequency: 6 issues per year
Published in: Biotechnic and Histochemistry, Volume 69, Issue 5 September 1994 ,
pages 289 - 290

Hi, Andrea,

I don't know of any microscopist that hasn't faced this problem at some time in their career if they have been working in this field for a long time. There are many potential causes for this problem. I don't know how many years you have been sectioning, so I don't want to insult you by going over basics. The first thing to do is to take some old blocks that did section well and cut them to check your microtome and knives to see if they are the source of the problem. Be aware of breezes in the room and room vibration while you are cutting, both of which can cause section thickness variation.
If you don't have these problems, and if your old blocks cut well, then you need to start to eliminate potential problems in your chemicals or processing procedure causing your current problems. Have you changed your processing schedule? If so, go back to your old procedure and see if the blocks cut well again. If you still have problems, then I would suspect a chemical problem. Is your alcohol or acetone fresh? If using propylene oxide, is it dry? Moisture in your dehydrating fluids can make for soft plastic, which will prevent your blocks from hardening properly, causing different cutting properties. Check your plastics. Are they old? Old plastics can absorb water from the air, which will affect their ability to harden, again affecting their cutting property. Check your oven temperature. Has it changed? Through this systematic process of elimination, you can track down the culprit. The key is to not change too many variables at one time, and to keep good notes of what you are changing. Humidity is a culprit that is ignored too often in processing tissue for E.M., and can play havoc with dehydrating fluids and plastic, as its effects are cumulative, a little more water building up in your fluids and plastics with each time a bottle is opened and left exposed to the lab room air. If your old blocks cut well, try fresh chemicals and see if water build-up in your chemicals isn't the problem that you are having. If this is the case, you may be able to "rescue" your existing tissue by hardening the blocks a little longer at a slightly higher temperature in your oven over the weekend and trying cutting them again. I routinely polymerize my Epon-equivalent (LX112) at 70 degrees C at least overnight, and often over the weekend, before cutting. In the past I have had to rescue batches of blocks when some of my plastic had picked up water and I was unaware of it (in my early days:) ). Sometimes it works, sometimes it doesn't, but it's worth a try. Now, I store all of my plastics in a Bell jar with desiccan!
t in it
to keep them dry, and work with small volumes of dehydrating fluids so my stock solutions are fresh. Working in Florida (read HUMID!!!) forces one to learn good technique :).

Ed Haller

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-cas.usf.edu
________________________________________
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Email: kaszas.1-at-osu.edu
Name: Andrea Kaszas

Organization: Ohio State University/OARDC

Title-Subject: [Filtered] sections thickness problem

Message: Dear Microscopist,

I have problems with the sectioning thickness nowadays. We have two
Leica microtomes. When we are sectioning thick or thin sections one
of the section is thicker and the second one is much thinner after
this a thicker section come again and a thinner after that and so on.
Have you ever been this kind of problem before? What can cause that problem?

Any suggestion would be greatly appreciated.

Thank you for your help in advance.

Andrea Kaszas

microscope technician

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From: kenconverse-at-qualityimages.biz
Date: Fri, 3 Sep 2010 07:58:29 -0500
Subject: [Microscopy] viaWWW: GW Electronics

Contents Retrieved from Microscopy Listserver Archives
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Clarissa,
My bad! You could also contact

Mike Dufraine
EM Product Manager
Energy Beam Sciences Inc
29-B Kripes Road
East Granby
CT 06026
USA

as he is listed as the US dealer for K E Developments.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


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Email: cvierret-at-mst.edu
Name: Clarissa Wisner

Organization: Missouri Universityof Science and Technology

Title-Subject: [Filtered] GW Electronics

Message: Greetings,

This is a two part query:

1. Does anybody have a current contact for the company GW
Electronics out of Georgia?

2. Does anybody know of a company in the US that can work on GW
chamberscopes?

Our chamberscope has a vacuum leak and we would like to have it
repaired versus replacing it.

Thanks in advance for your replies

Clarissa

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From: mdufraine-at-ebsciences.com
Date: Fri, 3 Sep 2010 08:10:32 -0500
Subject: [Microscopy] viaWWW: GW Electronics

Contents Retrieved from Microscopy Listserver Archives
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Clarissa-

Energy Beam Sciences, Inc. is the US Distributor for the KE Development line of equipment.
You may contact me at the following to review any service questions on your GW equipment.

Mike Dufraine
Energy Beam Sciences, Inc.
29B Kripes Road
East Granby, CT 06026
TEL 800-992-9037 X340

Mike.

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Organization: Missouri Universityof Science and Technology

Title-Subject: [Filtered] GW Electronics

Message: Greetings,

This is a two part query:

1. Does anybody have a current contact for the company GW
Electronics out of Georgia?

2. Does anybody know of a company in the US that can work on GW chamberscopes?

Our chamberscope has a vacuum leak and we would like to have it
repaired versus replacing it.

Thanks in advance for your replies

Clarissa

Login Host: 131.151.110.200
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From: wtivol-at-verizon.net
Date: Fri, 3 Sep 2010 15:14:57 -0500
Subject: [Microscopy] EM: where to buy 50 mesh coated grids, Memory lane

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

Good morning, Paul,

Don't know for sure, but did I pick up that one of your parents did E.M.? My father did, and taught me the craft. Thank you for writing back. I was taught E.M. back in 1974 at Mellon Institute in Pittsburgh, Pennsylvania by some old timers started doing E.M. back in the '60's on RCA TEM's. They used to align the lenses in the colums on those scopes with rubber mallets! The RCA's could only shoot 3 photos on a glass plate at a time, then you had to open the column, take out the plate, put in another, pump the scope down and take more photos. I came on just as the lab was switching from glass plates to sheet film in their other scopes, and as they were switching from using their two RCA's to using a Philips EM 200, Philips EM 300 (I got to use that when I was lucky), and a JEOL JEM 7.
In that lab in the Biology Department, they used to make holey carbon grids with ethylene dichloride and formvar. The tech there discovered that by chewing Wrigley's Spearmint gum she got different sized holes in the film when she breathed on the film then when she breathed on it without the gum. I wonder if someone with more time on their hands than myself would want to study the effect of chewing gum mint oils on hole formation in formvar films?

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-cas.usf.edu
________________________________________
X-from: paul r hazelton [paul_hazelton-at-umanitoba.ca]
Sent: Friday, September 03, 2010 1:47 AM
To: Haller, Edward
Cc: Microscopy Listserver


On Sep 3, 2010, at 6:20 AM, ehaller-at-health.usf.edu wrote:

} In that lab in the Biology Department, they used to make holey
} carbon grids with ethylene dichloride and formvar. The tech there
} discovered that by chewing Wrigley's Spearmint gum she got different
} sized holes in the film when she breathed on the film then when she
} breathed on it without the gum. I wonder if someone with more time
} on their hands than myself would want to study the effect of chewing
} gum mint oils on hole formation in formvar films?


Dear Ed,
Since gum chewing promotes salivation, an alternative explanation for
the difference in hole formation is that higher humidity of the breath
is responsible. The proposed experiment should, then, include Juicy
Fruit gum, and old vs. new gum.
Yours,
Bill


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From: PhillipsT-at-missouri.edu
Date: Tue, 7 Sep 2010 10:30:29 -0500
Subject: [Microscopy] Lensfree microscopy on a cellphone

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

Hi

As always happens when we are given more detail new critical areas come into
play!

I must first apologise to your service technician as with the extra
information I too believe it to be a problem with the gun alignment system.
The change from gun tilt into a gun shift mode suggests that this circuit
was not fully operational at that time. The movement of the instrument and
the changes when it is out of use all point in the defection coil direction.

I just wish the service technician good luck and hope the system crashes as
a very irregular fault like this is almost impossible to track down unless
the problem is present when the service technician is present.

Regards

Steve


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: greggps-at-umich.edu [mailto:greggps-at-umich.edu]
Sent: 02 September 2010 22:18
To: protrain-at-emcourses.com

Thanks to all who have responded. I really appreciate it.

I'd like to add a few details and see if they clarify things at all:

- I've changed out filaments, and the problem doesn't seem to be changed.
- The back and forth dancing of the beam was only present one or two days
over the past 4-5 months, so that might have been charging.
- Mostly, the beam just occasionally moves while the scope is not actively
being used.
- The physical Gun settings change in the display parameters.
- At one point, the Gun-Tilt-Y knob was actually moving the beam instead of
intensifying the illumination. As of this morning, this had temporarily
corrected itself. This was the detail that drove our engineer crazy, and
leads us to believe it's an electronic issue.
- There doesn't seem to be any unusual beam behavior when we change the
emission or the KV.
- Possibly unrelated: Our optimal Gun-Tilt settings are close to their
extreme limit and have been so since this EM unit was turned off for several
months, then moved to a new location and reactivated.

Do any two or more of these descriptions bring up any new theories?

Regards,
~Gregg

Gregg Sobocinski
Microscope Imaging Specialist
University of Michigan, MCDB Dept.
Ann Arbor, Michigan
USA

-----Original Message-----
X-from: greggps-at-umich.edu [mailto:greggps-at-umich.edu]
Sent: Wednesday, September 01, 2010 2:57 PM
To: Sobocinski, Gregg

Our Philips CM100 BioTwin TEM has developed an electronic defect, which
causes the gun Y-axis setting to drift. Some days the beam is relatively
stable, some days you can see the beam 'jump' back and forth up to a
centimeter's distance on the viewing stage (moving once every second), and
some days we'll turn on the filament to find the beam completely missing,
requiring a reasonable amount of alignment (not just the "Y" direction.)

Our service provider has not seen this previously, and has moved around and
tested the circuit boards with no effect on the problem. His last two
guesses are expensive (Probably $4,000-$7,000, and we've already put quite a
bit into troubleshooting and power supplies this year), so I wanted to know
if anyone else has encountered this phenomenon, and how you fixed it.

If we could confidently choose which item is defective, I can probably
budget for it, but those holding the purse strings are a little nervous
right now.

Your input is greatly appreciated. All suggestions or guesses are welcome.  
~Gregg

Gregg Sobocinski
Microscope Imaging Specialist
University of Michigan, MCDB Dept.
Ann Arbor, Michigan
USA




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From angelika.mueller-at-scaspach.de Sat Sep 4 16:39:10 2010
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Email: tomw-at-uidaho.edu
Name: Tom Williams

Organization: Univ of Idaho/Electron Microscopy Center

Title-Subject: [Filtered] Rocking Curve-XRD

Message: Question a little outside microscopy: I have been asked to
do a rocking curve analysis with my D5000 XRD to measure the mosaic
angle in Graphene/High-Order Pyrolytic Graphite. Embarrassed to say
that I have never performed a rocking curve! Would anyone have a
procedure, a good reference, tips, advice, or sage wisdom?

Thanks,
Tom

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I saw an interesting article highlighted by Faculty of 1000 that I thought I would share with the list - "Lensfree microscopy on a cellphone" Derek Tseng, Onur Mudanyali, Cetin Oztoprak, Serhan O. Isikman, Ikbal Sencan, Oguzhan Yaglidere and Aydogan Ozcan Lab Chip, 2010, 10, 1787-1792.

http://www.ncbi.nlm.nih.gov/sites/entrez/20445943?dopt=Abstract&holding=f1000,f1000m,isrctnumochsclib&tool=f1000tool



Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/



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From: greggps-at-umich.edu
Date: Tue, 7 Sep 2010 13:04:00 -0500
Subject: [Microscopy] Troubleshooting: TEM gun-Y drift. Need help with

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Quoted text: "I just wish the service technician good luck and hope the system crashes as
a very irregular fault like this is almost impossible to track down unless
the problem is present when the service technician is present."

Steve,
You are SO VERY correct, and have summed up the bane of computerized instruments!

I appreciate those thoughts and your insights. I feel better knowing we are not all so crazy with our diagnosis.

I'm anticipating an interesting few years until the problem gets significantly worse. In the meantime, I'm learning more and more tricks to recover from the malfunction more quickly.

Regards,
~Gregg

-----Original Message-----
X-from: Steve Chapman [mailto:protrain-at-emcourses.com]
Sent: Saturday, September 04, 2010 2:03 PM
To: Sobocinski, Gregg
Cc: Microscopical Soc of America

Thanks to all who have responded. I really appreciate it.

I'd like to add a few details and see if they clarify things at all:

- I've changed out filaments, and the problem doesn't seem to be changed.
- The back and forth dancing of the beam was only present one or two days
over the past 4-5 months, so that might have been charging.
- Mostly, the beam just occasionally moves while the scope is not actively
being used.
- The physical Gun settings change in the display parameters.
- At one point, the Gun-Tilt-Y knob was actually moving the beam instead of
intensifying the illumination. As of this morning, this had temporarily
corrected itself. This was the detail that drove our engineer crazy, and
leads us to believe it's an electronic issue.
- There doesn't seem to be any unusual beam behavior when we change the
emission or the KV.
- Possibly unrelated: Our optimal Gun-Tilt settings are close to their
extreme limit and have been so since this EM unit was turned off for several
months, then moved to a new location and reactivated.

Do any two or more of these descriptions bring up any new theories?

Regards,
~Gregg

Gregg Sobocinski
Microscope Imaging Specialist
University of Michigan, MCDB Dept.
Ann Arbor, Michigan
USA

-----Original Message-----
X-from: greggps-at-umich.edu [mailto:greggps-at-umich.edu]
Sent: Wednesday, September 01, 2010 2:57 PM
To: Sobocinski, Gregg

Our Philips CM100 BioTwin TEM has developed an electronic defect, which
causes the gun Y-axis setting to drift. Some days the beam is relatively
stable, some days you can see the beam 'jump' back and forth up to a
centimeter's distance on the viewing stage (moving once every second), and
some days we'll turn on the filament to find the beam completely missing,
requiring a reasonable amount of alignment (not just the "Y" direction.)

Our service provider has not seen this previously, and has moved around and
tested the circuit boards with no effect on the problem. His last two
guesses are expensive (Probably $4,000-$7,000, and we've already put quite a
bit into troubleshooting and power supplies this year), so I wanted to know
if anyone else has encountered this phenomenon, and how you fixed it.

If we could confidently choose which item is defective, I can probably
budget for it, but those holding the purse strings are a little nervous
right now.

Your input is greatly appreciated. All suggestions or guesses are welcome.  
~Gregg

Gregg Sobocinski
Microscope Imaging Specialist
University of Michigan, MCDB Dept.
Ann Arbor, Michigan
USA




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From: jkrupp-at-deltacollege.edu
Date: Tue, 7 Sep 2010 13:36:45 -0500
Subject: [Microscopy] FW: Troubleshooting: TEM gun-Y drift. - Response

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} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
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}
} Our Philips CM100 BioTwin TEM has developed an electronic defect,
} which
} causes the gun Y-axis setting to drift. Some days the beam is
} relatively
} stable, some days you can see the beam 'jump' back and forth up to a
} centimeter's distance on the viewing stage (moving once every
} second), and
} some days we'll turn on the filament to find the beam completely
} missing,
} requiring a reasonable amount of alignment (not just the "Y"
} direction.)
}

We had a similar problem on an old JEOL 100B. After a move, some of
the alignment controls did not work right. Checked all the boards,
pots, etc. All was well. Had to go through all kinds of gyrations to
make the thing work. Defect was intermittent, sometimes there and
sometimes not.

Turns out there was a short in the cable feed through for the
alignment controls. Depending on what direction the stress came on the
cable, it would work right or not. We think the movers may have seen
the cable as a nice 'handle' to help them with the move and cracked
the pin connections. From then on, we always remove the 'handles'
before letting the movers do their thing.

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu




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From: Henrik.Kaker-at-guest.arnes.si
Date: Tue, 7 Sep 2010 17:13:18 -0500
Subject: [Microscopy] viaWWW: Rocking Curve-XRD

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On Sep 6, 2010, at 1:10 PM, tomw-at-uidaho.edu wrote:

}
} Email: tomw-at-uidaho.edu
} Name: Tom Williams
}
} Organization: Univ of Idaho/Electron Microscopy Center
}
} Title-Subject: [Filtered] Rocking Curve-XRD
}
} Message: Question a little outside microscopy: I have been asked to
} do a rocking curve analysis with my D5000 XRD to measure the mosaic
} angle in Graphene/High-Order Pyrolytic Graphite. Embarrassed to say
} that I have never performed a rocking curve! Would anyone have a
} procedure, a good reference, tips, advice, or sage wisdom?
}
} Thanks,
} Tom
}

Dear Tom,
You're not too far from microscopy, since the rocking curve is the
same for electron diffraction (ED) as for XRD. The curve is simply
the intensity as a function of the angular displacement from the Bragg
condition. I have no experience analyzing it to determine mosaic
angle(s), but to obtain the curve, just measure the intensities of the
appropriate spot(s) for different orientations. It is likely that you
will need angular displacements in two dimensions; i.e., at different
orientations in both theta and phi (and combinations of these). I
don't know how far from the Bragg angle the curve should extend for
XRD; for ED it is only a degree or two, since the wavelength of the
electron is very short. I hope someone else has a reference; have you
tried Wikipedia?
Yours,
Bill


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Tom,

Please ask at XRD Liserver at
https://www.jiscmail.ac.uk/cgi-bin/webadmin?A0=XRD.

Henrik

On 7.9.2010 23:10, wtivol-at-verizon.net wrote:
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} On Sep 6, 2010, at 1:10 PM, tomw-at-uidaho.edu wrote:
}
} } Email: tomw-at-uidaho.edu
} } Name: Tom Williams
} }
} } Organization: Univ of Idaho/Electron Microscopy Center
} }
} } Title-Subject: [Filtered] Rocking Curve-XRD
} }
} } Message: Question a little outside microscopy: I have been asked to
} } do a rocking curve analysis with my D5000 XRD to measure the mosaic
} } angle in Graphene/High-Order Pyrolytic Graphite. Embarrassed to say
} } that I have never performed a rocking curve! Would anyone have a
} } procedure, a good reference, tips, advice, or sage wisdom?
} }
} } Thanks,
} } Tom
} }
} Dear Tom,
} You're not too far from microscopy, since the rocking curve is the
} same for electron diffraction (ED) as for XRD. The curve is simply
} the intensity as a function of the angular displacement from the Bragg
} condition. I have no experience analyzing it to determine mosaic
} angle(s), but to obtain the curve, just measure the intensities of the
} appropriate spot(s) for different orientations. It is likely that you
} will need angular displacements in two dimensions; i.e., at different
} orientations in both theta and phi (and combinations of these). I
} don't know how far from the Bragg angle the curve should extend for
} XRD; for ED it is only a degree or two, since the wavelength of the
} electron is very short. I hope someone else has a reference; have you
} tried Wikipedia?
} Yours,
} Bill
}
}
} ==============================Original Headers==============================
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--
Dr. Henrik Kaker
Ob Suhi 23
SI-2390 Ravne
Slovenia
GSM: +386 31 784 281
http://www.kaker.com
Email:info-at-kaker.com


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From: nizets2-at-yahoo.com
Date: Wed, 8 Sep 2010 09:09:14 -0500
Subject: [Microscopy] FIB/SEM applications in biology

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

Almost 2 months ago, I asked for help and would like to summerize the
replies. The majority suggested Grant Scientific Corporation
(http://www.grantscientific.com/) . Also some suggested EMS and SPI. In
fact, a google search shows that all the major em supply houses will
recoat or supply replacements.

In the end, I got mine from Grant as well and have been very happy with
it. The screens are bright with even illumination. The calibration marks
in gold show up better than the original black ones. Thanks for all that
replied.

On Mon, 12 Jul 2010, wpchan-at-u.washington.edu wrote:

} Hi
}
} What's your favorite vendor for recoating fluorescent viewing screen for a
} TEM? Thanks.

--
Pang (Wai Pang Chan, wpchan-at-u.washington.edu, PAB A087, 206-685-1519)
The Biology Imaging Facility (http://depts.washington.edu/if/)

==============================Original Headers==============================
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From ahamid-at-jointcommission.org Tue Sep 7 23:27:21 2010
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Dear colleagues,

Sadly I was not present in the M&M 2010 and I suppose that there was some talk
about the applications of FIB/SEM in biology.
I would like to gain some insight about this technique:
1) What is the thickness of the layer which can be etched by FIB on
Epoxy-embedded samples?
2) Is there a "lost thickness"? I mean, if the etching thickness is bigger than
the imaging thickness, there is a loss of material and then a loss of
informations.
3) Given that the imaging is only conceivable with BSE, what realistic XY
resolution is achievable?
4) Are there specific treatments, esp. with heavy metals, required to improve
the contrast of intracellular structures?
5) Is it realistically only applicable to tissues or is it also applicable to
single cell analysis?

More specifically, I would like to know if the presence of sub-micron particles
in cells can be detected and their localisation with respect to the cell 
membrane and the cell nucleus (or better if possible) be determined. The purpose
would be mainly to quantify the particles in the cells and get a good
approximation of their size distribution into the cells.
If sub-micron particles are present in the cells, would they be etched at
exactly the same depth as the rest of the cells (mainly light elements embedded
in Epoxy resin)? It is conceivable that the nature of the particles is important
to answer this question, I don't know enough about FIB to answer that.

I would like to mainly concentrate on Epoxy-embedded samples, not really on
cryo-substituted samples but if it can bring useful informations feel free to
discuss whichever preparation technique you like.

Thanks in advance and best regards.

Stephane





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From: ALawrence-at-entomology.msstate.edu
Date: Wed, 8 Sep 2010 12:13:37 -0500
Subject: [Microscopy] seeking an ultramicrotome

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

A university faculty member requested this be posted. Please contact him directly (off-line).

Seeking:

An ultramicrotome that is in good working form. It does not have to be anything really fancy or motorized. It is for sectioning of samples embedded in plastic for both plant and animal developmental biology research. Should be able to obtain sections that are 0.7-2 um range in thickness. The machine would be used in light microscopy applications (not TEM). All of the working parts should be present. Examples include but are not limited to:

SORVALL MT 5000 ULTRAMICROTOME W/ MICROSCOPE
SORVALL MT2 Ultramictrtome
SORVALL Model: MT2B
Ivan Sorvall JB-4 Microtome

Vincent Klink
Department of Biological Sciences
Mississippi State University
Mississippi State, MS 39762

vklink-at-biology.msstate.edu

Vincent Klink, Ph.D.
Department of Biological Sciences
Harned Hall; Rm 212 (office), 310 (lab)
Mississippi State University
Mississippi State, MS 39762

Phone: 662.325.4577
Fax: 662.325.7939




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From: LettJ-at-ent.wustl.edu
Date: Wed, 8 Sep 2010 15:21:06 -0500
Subject: [Microscopy] TEM need stable-storing uranyl and lead stains

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

Does anyone have any protocols for making uranyl acetate and,
especially, lead grid stains that will reliably store for several weeks?
Sometimes we go for quite a spell between needing to stain grids, and
end up needlessly wasting batches of stain solution. I'd like to make
up a batch of each and be able to store aliquots, if possible. (We're
currently using 5% Uranyl Acetate (aq) and Reynolds' Lead Citrate, but
are open to others.)

Thank you in advance,

Jaci

Jaclynn Lett
Senior Research Technician, EM Facility
Research Center for Auditory and Vestibular Studies
Department of Otolaryngology
Washington University School of Medicine
660 S. Euclid Ave., Campus Box 8115
St. Louis, MO 63110
Office: 314-747-7257
Fax: 314-747-7230
Email: lettj-at-ent.wustl.edu
Website: http://otocore.wustl.edu


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From: lene.cecilie.hermansen-at-nvh.no
Date: Wed, 8 Sep 2010 17:50:24 -0500
Subject: [Microscopy] viaWWW:looking for images of Artemia

Contents Retrieved from Microscopy Listserver Archives
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I make up 10 ml at a time and store each one in a 10ml syringe, fitted
with a 0.22 micron filter and cap. I cover the uranyl acetate syringe and
filter with aluminum foil. I store them at room temperature and they last
for weeks to months. When I'm using them I discard the first couple of
drops out of the end, figuring it was on the wrong side of the filter.
This has worked great for me for decades!

Aloha,
Tina

} Does anyone have any protocols for making uranyl acetate and,
} especially, lead grid stains that will reliably store for several weeks?
} Sometimes we go for quite a spell between needing to stain grids, and
} end up needlessly wasting batches of stain solution. I'd like to make
} up a batch of each and be able to store aliquots, if possible. (We're
} currently using 5% Uranyl Acetate (aq) and Reynolds' Lead Citrate, but
} are open to others.)
}
} Thank you in advance,
}
} Jaci
}
} Jaclynn Lett
} Senior Research Technician, EM Facility
} Research Center for Auditory and Vestibular Studies
} Department of Otolaryngology
} Washington University School of Medicine
} 660 S. Euclid Ave., Campus Box 8115
} St. Louis, MO 63110
} Office: 314-747-7257
} Fax: 314-747-7230
} Email: lettj-at-ent.wustl.edu
} Website: http://otocore.wustl.edu
}
}
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****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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From arto.juva-at-aj-consultants.com Wed Sep 8 17:19:25 2010
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Received: from [200.27.169.170] by mq01.mail.saunalahti.fi; Wed, 8 Sep 2010 18:15:32 -0400

Jaci,

Uranyl acetate will last for several months if it is made in water and kept
in the refrigerator. I have been using 2% for over 30 years but I do use a
block stain of 1% UA before dehydration of the specimens. There is a slight
sediment in the bottom of the bottle but I only take from the top and
discard that in our mixed waste when I make a new batch. When I am ready to
stain I put a drop of 2% UA on parafilm inside a petri dish and add a drop
of 100% Ethanol, a grid and close the dish which has been covered to keep
out the light. Ten minutes and it is ready to be water washed a few times
then left in a drop of water in dish two before putting the grid into lead
stain (no drying).

Lead Stain
X-from the 25th M&M meeting in Chicago in 1967.
Ref. is on page 148-149. By Aly Fahmy, M.D. Ph.D.
50ml water boiled and cooled into a 50 ml Polypropylene test tube
1 dry NaOH pellet into the water, cap the tube and dissolve pellet
completely. Add 0.2 to 0.25 grams lead citrate (tip of spatula)
Invert tube until all is dissolved and clear.
Let sit 15 min. in the dark for crystals to settle (if any). This works as
well as Reynolds' Lead Citrate for me.

Fill syringes (1 to 10ml), attach an 18 gauge needle, expel air
and stick into a large rubber stopper. Discard the last few drops left in
the tube. Label with date.
Put into the back of a refrigerator. These can last a year but as a rule I
discard them after 6 months. Take out one at a time to use at room
temperature. Stick the cold syringe into a smaller stopper in a dark cabinet
or put a tall metal can over it and allow time to warm-up. Expel the first
few drops incase of crystals or use a small syringe filter. Note: some 0.2
micron filters do not work ­ the filtered stain will not stain!
The syringe 'in use' is usually fine for a week or so if kept dark.

I put out a few drops at a time on fresh parafilm in a third dish and stain
for 1 to 2 minutes. Wash well and dry.
FYI Excessive lead staining time will cause the stain to lighten, not get
better.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E ­ Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-mail.nih.gov

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and can be freely shared
and reproduced.
============

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Email: lene.cecilie.hermansen-at-nvh.no
Name: Lene Cecilie Hermansen

Organization: The Norwegian School of Veterinary Science

Title-Subject: [Filtered] Artemia

Message: Hi
I have a group that wants to look at the gut of Artemia. They are
very inexperienced with morphology, at least at this stage, and I
wonder if anybody has any pictures they can have a look at.

Thanks!

Regards Lene Hermansen,
The Norwegian School of Veterinary Science


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From: cemn.physics-at-gmail.com
Date: Wed, 8 Sep 2010 17:51:44 -0500
Subject: [Microscopy] viaWWW: Materials Microstructure and Microanalysis Workshop

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Email: cemn.physics-at-gmail.com
Name: Zhiqiang Chen

Organization: Portland State University

Title-Subject: [Filtered] Materials Microstructure and Microanalysis Workshop

Message: Materials Microstructure and Microanalysis
Workshop

October 18~20th 2010

1025 SW Mill St, Science Building One Room 22 Portland OR 97201

The Center for Electron Microscopy and
Nanofabrication -at- Portland State University
cordially invites you to attend 3-day Materials
Microstructure and Microanalysis Workshop
including lectures and hands-on practice of
different microscopy techniques. Topics will
include:
ï Analytical TEM, including (S)TEM/BF/DF
imaging and EDX and EEL spectroscopies
ï Analytical SEM, including BSE/CL imaging and EDX and WDX spectrocopies
ï Dual Beam FIB, including TEM specimen
preparation, serial sectioning and EDX
spectroscopy

Technical tutorial lectures on general
microstructure and microanalysis theory will be
given by FEI, Oxford Instrument, Gatan, Zeiss
application engineers, and PSU faculty each
morning, followed by TEM, SEM, and FIB practice
sessions each afternoon of October 18th~20th 2010.

Registration DEADLINES: October 1st, 2010

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From: RHBerg-at-danforthcenter.org
Date: Wed, 8 Sep 2010 17:59:27 -0500
Subject: [Microscopy] Re: TEM need stable-storing uranyl and lead stains

Contents Retrieved from Microscopy Listserver Archives
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Jaci,

Sato's lead is remarkably stable--I am still using the stock I made in
November, 2008. This link has recipes : http://www.2spi.com/catalog/chem/sato_recipe.html


Cheers,

Howard



R. Howard Berg, Ph.D.
Director, Integrated Microscopy Facility
Danforth Plant Science Center
975 N. Warson Rd.
St. Louis, MO 63132

ph 314-587-1261 fx 314-587-1361 cell 314-378-2409
rhberg-at-danforthcenter.org www.danforthcenter.org
visit this educational resource: http://www.danforthcenter.org/science/core_facilities/integrated_microscopy/gallery.asp








On Sep 8, 2010, at 3:22 PM, LettJ-at-ent.wustl.edu wrote:

}
}
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} Does anyone have any protocols for making uranyl acetate and,
} especially, lead grid stains that will reliably store for several
} weeks?
} Sometimes we go for quite a spell between needing to stain grids, and
} end up needlessly wasting batches of stain solution. I'd like to make
} up a batch of each and be able to store aliquots, if possible. (We're
} currently using 5% Uranyl Acetate (aq) and Reynolds' Lead Citrate, but
} are open to others.)
}
} Thank you in advance,
}
} Jaci
}
} Jaclynn Lett
} Senior Research Technician, EM Facility
} Research Center for Auditory and Vestibular Studies
} Department of Otolaryngology
} Washington University School of Medicine
} 660 S. Euclid Ave., Campus Box 8115
} St. Louis, MO 63110
} Office: 314-747-7257
} Fax: 314-747-7230
} Email: lettj-at-ent.wustl.edu
} Website: http://otocore.wustl.edu
}
}
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From: lene.cecilie.hermansen-at-nvh.no
Date: Thu, 9 Sep 2010 08:03:47 -0500
Subject: [Microscopy] viaWWW: Transmission EM, Artemia

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Name: Lene Cecilie Hermansen

Organization: The Norwegian School of Veterinary Science

Title-Subject: [Filtered] Transmission EM, Artemia

Message: I am sorry; but in my last posting I forgot to mention that
I am looking for transmission electron microscopy pictures of Artemia.

Last posting:
Hi

I have a group that wants to look at the gut of Artemia. They are very
inexperienced with morphology, at least at this stage, and I wonder if
anybody has any pictures they can have a look at.

Thanks!

Regards Lene Hermansen,
The Norwegian School of Veterinary Science


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From: connellyps-at-nhlbi.nih.gov
Date: Thu, 9 Sep 2010 11:16:33 -0500
Subject: [Microscopy] JEM-1200EX TEM available

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I am posting this for our facility.
Please reply to Dr. Daniels as requested below.
Pat Connelly
==========

JEM-1200EX TEM available for transfer or donation in October

The NHLBI/NIH Electron Microscopy Core Facility will be replacing a
JEM-1200EX transmission EM with a current model, which is scheduled to be
delivered on October 18, 2010. The JEM-1200EX is 23 years old but it is in
good operating condition and has been under service contract with JEOL the
whole time. According to regulations, it must first be made available for
transfer to another NIH or other Federal lab, but if not claimed, it will be
available for donation to any non-profit research or educational institution
in the United States. The recipient of the instrument will be responsible
for transportation to their facility as well as installation and service
contract, if any.

The instrument and accessories are as originally purchased, including a
goniometer specimen stage with plus/minus 30 degrees tilt, a 2-place
standard specimen holder and 2-place rotation holder, a 3 1/4 by 4" film
camera that holds 50 cassettes, and a film dessicator.
We are currently using a bottom-mount AMT digital camera with this scope but
the camera will be transferred to another scope.

If you are potentially interested in acquiring this JEM-1200EX, please
contact me by e-mail. My address is danielsm2-at-mail.nih.gov.

Mathew P. Daniels, Ph.D.
Director - NHLBI Electron Microscopy Core Facility
National Institutes of Health
Bethesda, MD
danielsm2-at-mail.nih.gov



==============================Original Headers==============================
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From: hartfield-at-omniprobe.com
Date: Thu, 9 Sep 2010 11:45:01 -0500
Subject: [Microscopy] FIB/SEM applications in biology

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

Stephane,

There are some excellent recent articles that cover the type of information you seek. I outline 5 of these in my Aug. 24 blog post at info.omniprobe.com . There are different SEM imaging approaches, and answers to your questions depend on sample preparation and slicing methods, so a simple answer is not so easy. Initially applied for brain mapping and 3D imaging of tissue, it appears that as biological researchers adopt FIB-SEM strategies, the benefits are just now being realized and the application space is growing. This is a nascent field of exploration that is still in the early adoption phase and will be exciting to watch (and help) grow. While samples investigated by FIB-SEM include brain, blood vessels and DNA, US researchers at the NIH have recently shown its usefulness for larger cells and have used this to gain new insights into HIV transmission between dendritic cells and T-cells [PNAS 107(30): 13336-13341]. I leave further comments to the people actually doing this work.

Best Regards,
Cheryl
 -------------------------------------------------------------
Cheryl Hartfield, hartfield-at-omniprobe.com
Senior Applications Specialist, Omniprobe Inc.
Ph: (214) 572-6804; Fax: (214) 572-6801
http://www.omniprobe.com  


-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Wednesday, September 08, 2010 9:16 AM
To: Cheryl Hartfield

Dear colleagues,

Sadly I was not present in the M&M 2010 and I suppose that there was some talk
about the applications of FIB/SEM in biology.
I would like to gain some insight about this technique:
1) What is the thickness of the layer which can be etched by FIB on
Epoxy-embedded samples?
2) Is there a "lost thickness"? I mean, if the etching thickness is bigger than
the imaging thickness, there is a loss of material and then a loss of
informations.
3) Given that the imaging is only conceivable with BSE, what realistic XY
resolution is achievable?
4) Are there specific treatments, esp. with heavy metals, required to improve
the contrast of intracellular structures?
5) Is it realistically only applicable to tissues or is it also applicable to
single cell analysis?

More specifically, I would like to know if the presence of sub-micron particles
in cells can be detected and their localisation with respect to the cell 
membrane and the cell nucleus (or better if possible) be determined. The purpose
would be mainly to quantify the particles in the cells and get a good
approximation of their size distribution into the cells.
If sub-micron particles are present in the cells, would they be etched at
exactly the same depth as the rest of the cells (mainly light elements embedded
in Epoxy resin)? It is conceivable that the nature of the particles is important
to answer this question, I don't know enough about FIB to answer that.

I would like to mainly concentrate on Epoxy-embedded samples, not really on
cryo-substituted samples but if it can bring useful informations feel free to
discuss whichever preparation technique you like.

Thanks in advance and best regards.

Stephane





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==============================Original Headers==============================
21, 29 -- From hartfield-at-omniprobe.com Thu Sep 9 11:45:01 2010
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From: michael-at-shaffer.net
Date: Thu, 9 Sep 2010 13:40:00 -0500
Subject: [Microscopy] EPMA: mounting rare samples w/o waste

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

A researcher here just brought me some meteorite samples that he had paid
dearly for.  They have already been sectioned into slices but not mounted. 
As slices they are no thicker than 2mm, a couple only 1mm, while their
dimensions vary from 25 x 40mm to 5 x 5mm.  We want to mount them for EMPA
(& image analysis), but also reserve the possibility of also making thin
sections from them in the future (… ‘cept for maybe the 1mm thick slices). 
Before we go ahead and embed them into 30mm round epoxy, I thought someone
may be able to refer us to a technique designed around trying to accomplish
the same while wasting almost nothing(?)

… our need is almost immediate.

~~~~~~~~~~~~~~~~~~~~~
Cheerios, Michael Shaffer  :o)
SEM-MLA Research Coordinator
INCO Innovation Ctr, Memorial University
St. John's Newfoundland




==============================Original Headers==============================
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From: SWalck-at-SouthBayTech.com
Date: Thu, 9 Sep 2010 16:48:29 -0500
Subject: [Microscopy] EPMA: mounting rare samples w/o waste

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

Michael,

Recently, I've been working with a geologist on doing EBSD analysis using
petrographic samples and ion polishing with our IBS/e with KDC-10 system
over the entire sample (25x40mm). They had their samples prepared by an
outside lab. I have not been impressed with their mechanical polishing of
the samples that have both very hard and very soft phases in it and I think
that our lapping fixtures could do a better job. The samples have a lot of
surface relief because of the different phases and spending a lot of time
using abrasives with cloths.

You may want to take a look at our lapping fixtures and lapping machines.
(http://southbaytech.com/shop/mlp1.shtml and
http://southbaytech.com/shop/920.shtml) I would suggest the Model 155D or
Model 155DV. These are ideally suited for getting samples down for
petrographic analysis. If your EPMA can handle glass slides that are
approximately 2" along the diagonal. The "V" in the suffix of the fixture
designates vacuum. The glass slide can be put onto the mounting block just
using a vacuum. The petrographic thickness is usually about 30 um. You can
do that with our fixture. You can grind one side flat and then flip it over
and go down to the 30 um that you want. You can do the EPMA at that time.
If you like, I can send you a DVD on lapping that discusses it. The beauty
of using our system is that you can choose to do the EPMA first or do the
petrographic preparation first and then do the EPMA analysis can be done on
the same sample after coating the samples with a thin conductive coating.
For the studies that we were doing for EBSD, we put on a 10Å layer of Ir
with the IBS/e after the ion polishing and the samples did not charge using
a 30 keV beam. We used a high beam current (highest on the FEG), but not as
high as would be available in an EPMA. The 10 Å of Ir will not interfere
with your analysis.

Disclaimer: SBT manufactures and sells the lapping fixtures and IBS/e and
KDC-10 ion system.

-Scott

Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA 92673

eMail: SWalck-at-SouthBayTech.com
Phone: 949-492-2600
Toll Free: 1-800-728-2233 (USA)
Fax: 949-492-1499

www.SouthBayTech.com


-----Original Message-----
X-from: michael-at-shaffer.net [mailto:michael-at-shaffer.net]
Sent: Thursday, September 09, 2010 11:50 AM
To: swalck-at-southbaytech.com

A researcher here just brought me some meteorite samples that he had paid
dearly for.  They have already been sectioned into slices but not mounted. 
As slices they are no thicker than 2mm, a couple only 1mm, while their
dimensions vary from 25 x 40mm to 5 x 5mm.  We want to mount them for EMPA
(& image analysis), but also reserve the possibility of also making thin
sections from them in the future (… ‘cept for maybe the 1mm thick slices). 
Before we go ahead and embed them into 30mm round epoxy, I thought someone
may be able to refer us to a technique designed around trying to accomplish
the same while wasting almost nothing(?)

… our need is almost immediate.

~~~~~~~~~~~~~~~~~~~~~
Cheerios, Michael Shaffer  :o)
SEM-MLA Research Coordinator
INCO Innovation Ctr, Memorial University
St. John's Newfoundland




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==============================Original Headers==============================
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From: mike.bode-at-resaltatech.com
Date: Thu, 9 Sep 2010 18:07:47 -0500
Subject: [Microscopy] printers for lab

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


Hello Everybody,

Two weeks ago someone had asked about printers for the lab, and I had asked
to send me information so I can collect it and provide the information here.
I was hoping to get enough information so that this could be a resource for
everyone looking for a printer. Unfortunately, that was not the case. I
received 2 responses and I will just post these responses here (names
withheld).

If I get more responses I will follow-up again, but it seems that the
printing issue of images is not really the issue that keeps people up at
night.

X-from my own observation, I would say that the paper is one of the critical
elements. The better the paper, the better the results. Printing on everyday
office paper will not produce high quality results. My suggestion would be
to use a good quality laser printer with multiple trays in the lab. Print on
regular paper for just internal documentation, on high-gloss paper for
better prints. Inkjets can produce astonishing output, but they need to be
calibrated, are slower, and tend to clog. I am using an Epson R800 (older
model) for good results, but ever so often I have to throw away a cartridge
because it’s not working anymore, even though there is still ink in it. But
just like lasers, you get better results on photographic paper. I’d like to
hear about the current crop of dye-sub printers. They used to be better (and
more expensive), but I am not sure what the current printers deliver.

So, here are the two responses that I received:

“For my Hitachi S3000N SEM I have used HP inkjets all along.  When we got
the instrument in 1999 I ordered it with a Kodak dye sub printer which was
equivalent to a Codonics unit.  I had problems from the start with slow
printing times.  I tried an HP inkjet (forget the number now) and did some
side by side tests against the Kodak and got just as good results from the
HP with a lot less excitement.  So sadly the expensive Kodak has sat in a
corner of the lab ever since.

I don't have the issues with student use or need to worry a lot about duplex
printing that was mentioned.  I use an HP 6940 now with Epson matte finish
inkjet paper for routine SEM images and am very pleased with the output and
costs.  I can do prints on glossy photo paper if really necessary but day to
day the Epson paper works very well.”

And

“HP LaserJet 4700dn with HP glossy brochure paper. Printing profiles not
well supported so calibration is a pain, but the laser is quick and dirty
anyway. My users go to the photographic unit for the critical printing on
professional inkjet papers.”


---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com










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From: johnf-at-geology.wisc.edu
Date: Thu, 9 Sep 2010 20:47:10 -0500
Subject: [Microscopy] Galling of metal in a Wehnalt cap

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

In April while i was away from the lab, there was an incident with
our Hitachi S3400 VP-sem (W filament) where my assistant reported he
was unable to remove the old filament from the Wehnalt, and
eventually had to purchase another wehnalt. I figured someone cross
threaded the filament .....

However, today the same thing happened to me, and I could not remove
it. With a machinists help and good old Armstrong ingenuity, we
eventually were able to remove it after breaking off a bunch of Al
in the thread grooves. He told me about "galling" which this appears
to be and which he has had other experience with, where for whatever
reason, the metal surfaces apparently bond very strongly (whether
debris or asperities of the surfaces).

My question to you all---has anyone else seen this phenomenon with
Wenhalt caps? It's never happened in 18 years with our Cameca SX51,
and only after 4-5 years with the Hitachi has it happened.

John
--
========================================================
John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480
Cameron Electron Microprobe Lab lab: (608) 265-4798
Department of Geoscience fax: (608) 262-0693
University of Wisconsin home: (608) 274-2245
1215 West Dayton St. email: johnf-at-geology.wisc.edu
Madison, WI 53706 amateur radio: WA3BTA
Personal http://www.geology.wisc.edu/~johnf/
Probe lab http://www.geology.wisc.edu/~johnf/sx51.html
Probe Sign Up Calender:
http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi

"The first rule of all intelligent tinkering is to save every cog and
wheel." -- Aldo Leopold

"For a successful technology, reality must take precedence over
public relations, for Nature cannot be fooled." -- Richard P.
Feynman

==============================Original Headers==============================
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From: vitalylazar-at-att.net
Date: Thu, 9 Sep 2010 21:20:39 -0500
Subject: [Microscopy] Re: printers for lab

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

Hello listers,

I recommend the following printer/paper combination:

Printer - Brother HL-5370DW (average "street price" $200). Nice print
quality and nice GUI for bright/contrast/gamma adjustment with multiple
presets stored.

Paper - Hammermill OfficeOne Business Gloss by International Paper
company. Product # 16302-0. Costs about $8 per pack of 300 sheets. Paper
is acid-free. Brightness 92, weight 32 lb. Other (glossier) papers with
higher brightness numbers produced inferior prints. No idea why.

Above printer/paper combination produces prints of somewhat lesser
quality than photo inkjet (surprisingly not much), but the cost of
printing is at least 20 times cheaper.

The second best paper is Color Gloss, same brand, same price. Minor
difference in image contrast in favor of the former.

Actually Business Gloss paper tested better than any other paper on
laser printers of various brands over the years.

I agree about printing not being important at most places and in most
cases. We used to supply photo inkjet and laser printer (2 printers)
with every TEM camera system. Then switched to laser printers only. Then
to no printer at all unless specifically requested by end user. The
reason - most printers were never turned on.

Mike is right about paper. Paper is critically important. With this said
I advise against spending more than $300 on a laser printer for routine
everyday printing unless one has very good reason for doing so, because
(with some degree of generalization)-

- Printing unit is very similar if not identical throughout multiple
models of a given brand. I tested many laser printers in price range
from $100 to $2K+. If manufacturer got it right, then print quality of
$200 and $1K+ printers is identically good (on a good paper) within
given brand. If not - print quality will be identically less. Cost of a
laser printer reflects convenience and versatility rather than the print
quality. (networking, multiple trays for multiple paper formats, speed,
duty factor, etc.).

If your laser printer was $200 brand new and broke after 2 years and
8,000 prints you will junk it and buy another (better) one. I doubt you
do that with $3K printer that stopped working. Repairs, headaches...

Regarding estimating print quality by looking at printer specifications-
good luck with that... Nothing beats the test.


Vitaly Feingold
SIA
2773 Heath Lane
Duluth GA 30096
Ph. 770-232-7785
Fax 770-232-1791
www.sia-cam.com
vitaly-at-sia-cam.com

On 9/9/2010 7:09 PM, mike.bode-at-resaltatech.com wrote:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
}
} Hello Everybody,
}
} Two weeks ago someone had asked about printers for the lab, and I had asked
} to send me information so I can collect it and provide the information here.
} I was hoping to get enough information so that this could be a resource for
} everyone looking for a printer. Unfortunately, that was not the case. I
} received 2 responses and I will just post these responses here (names
} withheld).
}
} If I get more responses I will follow-up again, but it seems that the
} printing issue of images is not really the issue that keeps people up at
} night.
}
} X-from my own observation, I would say that the paper is one of the critical
} elements. The better the paper, the better the results. Printing on everyday
} office paper will not produce high quality results. My suggestion would be
} to use a good quality laser printer with multiple trays in the lab. Print on
} regular paper for just internal documentation, on high-gloss paper for
} better prints. Inkjets can produce astonishing output, but they need to be
} calibrated, are slower, and tend to clog. I am using an Epson R800 (older
} model) for good results, but ever so often I have to throw away a cartridge
} because it’s not working anymore, even though there is still ink in it. But
} just like lasers, you get better results on photographic paper. I’d like to
} hear about the current crop of dye-sub printers. They used to be better (and
} more expensive), but I am not sure what the current printers deliver.
}
} So, here are the two responses that I received:
}
} “For my Hitachi S3000N SEM I have used HP inkjets all along. When we got
} the instrument in 1999 I ordered it with a Kodak dye sub printer which was
} equivalent to a Codonics unit. I had problems from the start with slow
} printing times. I tried an HP inkjet (forget the number now) and did some
} side by side tests against the Kodak and got just as good results from the
} HP with a lot less excitement. So sadly the expensive Kodak has sat in a
} corner of the lab ever since.
}
} I don't have the issues with student use or need to worry a lot about duplex
} printing that was mentioned. I use an HP 6940 now with Epson matte finish
} inkjet paper for routine SEM images and am very pleased with the output and
} costs. I can do prints on glossy photo paper if really necessary but day to
} day the Epson paper works very well.”
}
} And
}
} “HP LaserJet 4700dn with HP glossy brochure paper. Printing profiles not
} well supported so calibration is a pain, but the laser is quick and dirty
} anyway. My users go to the photographic unit for the critical printing on
} professional inkjet papers.”
}
}
} ---
} Mike Bode, Ph.D.
} ResAlta Research Technologies Corp.
}
} 2102 Beech Ct
} Golden, CO 80401
} USA
}
} p: +1-303-748-4346
} f: +1-303-202-6350
} Mike.Bode-at-ResAltaTech.com
} www.ResAltaTech.com
}
}
}
}
}
}
}
}
}
}
} ==============================Original Headers==============================
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From: bigelow-at-umich.edu
Date: Fri, 10 Sep 2010 01:09:41 -0500
Subject: [Microscopy] Galling in grid cap

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

I have never experienced the problem John describes; however, galling
of threads can usually be prevented by lubricating the threads
lightly (thus, I conclude I never got my grid caps clean enough to
gall). In this situation the choice of a lubricant poses a serious
problem. It would, of course, be impossible to use any organic
lubricant such as a vacuum oil or grease. However, I wonder if it
might not be possible to use a solid lubricant such as molybdenum
disulfide, molybdenum diselenide, tungsten diselenide, etc. These
are all suitable lubricants for use in ordinary vacuum applications;
perhaps one of them might also withstand the temperatures that
prevail in the electron gun.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From: johnf-at-geology.wisc.edu
Date: Fri, 10 Sep 2010 07:27:45 -0500
Subject: [Microscopy] Galling of metal in a Wehnalt cap--Clarification

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

Clarification: only the filament assembly is Al ("pre centered
filament"). The Wenhalt itself is a steel, presumably stainless
steel. It is only 5 months old.

We have been having also a lot of problems with rather short
lifetimes of our filaments (purchased from Hitachi); for several
years we would get 100-200 hours out of a filament, but for the past
year or so we have been getting 30-50 hours per filament. Could there
be some cause-effect relationship with the galling issue?


}
}
} In April while i was away from the lab, there was an incident with
} our Hitachi S3400 VP-sem (W filament) where my assistant reported he
} was unable to remove the old filament from the Wehnalt, and
} eventually had to purchase another wehnalt. I figured someone cross
} threaded the filament .....
}
} However, today the same thing happened to me, and I could not remove
} it. With a machinists help and good old Armstrong ingenuity, we
} eventually were able to remove it after breaking off a bunch of Al
} in the thread grooves. He told me about "galling" which this appears
} to be and which he has had other experience with, where for whatever
} reason, the metal surfaces apparently bond very strongly (whether
} debris or asperities of the surfaces).
}
} My question to you all---has anyone else seen this phenomenon with
} Wenhalt caps? It's never happened in 18 years with our Cameca SX51,
} and only after 4-5 years with the Hitachi has it happened.
}
} John
} --
} ========================================================
} John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480
} Cameron Electron Microprobe Lab lab: (608) 265-4798
} Department of Geoscience fax: (608) 262-0693
} University of Wisconsin home: (608) 274-2245
} 1215 West Dayton St. email: johnf-at-geology.wisc.edu
} Madison, WI 53706 amateur radio: WA3BTA
} Personal http://www.geology.wisc.edu/~johnf/
} Probe lab http://www.geology.wisc.edu/~johnf/sx51.html
} Probe Sign Up Calender:
} http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi
}
} "The first rule of all intelligent tinkering is to save every cog and
} wheel." -- Aldo Leopold
}
} "For a successful technology, reality must take precedence over
} public relations, for Nature cannot be fooled." -- Richard P.
} Feynman
}
} ==============================Original Headers==============================
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--
========================================================
John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480
Cameron Electron Microprobe Lab lab: (608) 265-4798
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wheel." -- Aldo Leopold

"For a successful technology, reality must take precedence over
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Feynman

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8, 24 -- From johnf-at-geology.wisc.edu Fri Sep 10 07:27:45 2010
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From: gregory.a.jerman-at-nasa.gov
Date: Fri, 10 Sep 2010 08:25:50 -0500
Subject: [Microscopy] Re: Galling of metal in a Wehnalt cap

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

The galling you are experiencing is a result of the wehnelt cap threads
being welded to the base threads. We have a Cameca SX-50 that welded two
LaB6 wehnalts because they got too hot. Normally the tungsten wehnelts
don't get hot enough to weld the threads, but your filament life issue
indicates you are running very hot. We have a Hitachi S-3700N which I
believe has the same gun as your S-3400. The preset filaments should come
with small copper gaskets. Make sure at least one is in place between the
wehnelt cap and the base. More gaskets will improve your filament life at
the expense of beam brightness. Also make sure that the filament setting is
in the mid-80's. We have had trouble in the past with the software randomly
pegging the filament setting to 100 which way over saturates the filament
and makes it run hot. We have burnt out a number of filaments quickly
because of this, but have never welded the wehnelt cap.

Greg

Gregory A. Jerman
NASA MSFC
EM31 Materials Diagnostics Team
gregory.a.jerman-at-nasa.gov




} From: "johnf-at-geology.wisc.edu" {johnf-at-geology.wisc.edu}
} Reply-To: "johnf-at-geology.wisc.edu" {johnf-at-geology.wisc.edu}
} Date: Thu, 9 Sep 2010 20:50:45 -0500
} To: "Jerman, Gregory A. (MSFC-EM31)" {gregory.a.jerman-at-nasa.gov}
} Subject: [Microscopy] Galling of metal in a Wehnalt cap
}
}
}
}
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} In April while i was away from the lab, there was an incident with
} our Hitachi S3400 VP-sem (W filament) where my assistant reported he
} was unable to remove the old filament from the Wehnalt, and
} eventually had to purchase another wehnalt. I figured someone cross
} threaded the filament .....
}
} However, today the same thing happened to me, and I could not remove
} it. With a machinists help and good old Armstrong ingenuity, we
} eventually were able to remove it after breaking off a bunch of Al
} in the thread grooves. He told me about "galling" which this appears
} to be and which he has had other experience with, where for whatever
} reason, the metal surfaces apparently bond very strongly (whether
} debris or asperities of the surfaces).
}
} My question to you all---has anyone else seen this phenomenon with
} Wenhalt caps? It's never happened in 18 years with our Cameca SX51,
} and only after 4-5 years with the Hitachi has it happened.
}
} John
} --
} ========================================================
} John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480
} Cameron Electron Microprobe Lab lab: (608) 265-4798
} Department of Geoscience fax: (608) 262-0693
} University of Wisconsin home: (608) 274-2245
} 1215 West Dayton St. email: johnf-at-geology.wisc.edu
} Madison, WI 53706 amateur radio: WA3BTA
} Personal http://www.geology.wisc.edu/~johnf/
} Probe lab http://www.geology.wisc.edu/~johnf/sx51.html
} Probe Sign Up Calender:
} http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi
}
} "The first rule of all intelligent tinkering is to save every cog and
} wheel." -- Aldo Leopold
}
} "For a successful technology, reality must take precedence over
} public relations, for Nature cannot be fooled." -- Richard P.
} Feynman
}
} ==============================Original Headers==============================
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From: dufresne-at-ms.umanitoba.ca
Date: Sun, 12 Sep 2010 09:49:41 -0500
Subject: [Microscopy] viaWWW: Nano Particles Imaging

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

As a biologist, my experience with 'galling' has been confined to interpersonal conflict, horses and their saddles, etc. I have never even been aware that anything in the microscopes I attend to each day might be 'galling'. Thus, in haste, I searched for a 'material' definition/explanation of what all galls non-biologics.

Having found some brief success in my search, I sought out a deeper understanding of the nuts and bolts of the matter.

I found it here (below), and thought it might help other non-materialists with a similar vexing problem with 'galling'.

http://www.estainlesssteel.com/gallingofstainless.html

Cheers to all, and may all of your 'gallings' be miner.

Fred Monson

Frederick C. Monson, PhD
Tecnhical Director
Center for Microanalysis and Imaging Research and Training Center (CMIRT)
Large Scientific Instrument Core
West Chester University S. Church St. and W. Rosedale Ave.
West Chester, PA, 19383
610-738-0437

Frederick C. Monson, PhD
Tecnhical Director
Center for Microanalysis and Imaging Research and Training Center (CMIRT)
Large Scientific Instrument Core
West Chester University S. Church St. and W. Rosedale Ave.
West Chester, PA, 19383
610-738-0437
fmonson-at-wcupa.edu

Reads of the Month:
1. "The Liver as a Lymphoid Organ," Ian Nicholas Crispe, Annual Review of Immunology, Apr 2009, Vol. 27: 147-163.
2. "A Personal Journey of Discovery: Developing Technology and Changing Biology," Lee Hood, Annual Review of Analytic Chemistry, Vol. 1: 1-43.
3. "On the Biomechanics of Vaginal Birth and Common Sequelae", James A. Ashton-Miller1 and John O.L. DeLancey, Annual Review Biomed. Eng, 11: 163-176.(2009)

New Web Site: http://cmirt.wcupa.edu/index.html
New Scheduler: http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl
=========================================================================
History is Cyclic. Only the ism's, nouns or adjectives change, AND......, When the objective classroom turns to agenda, history repeats!
=========================================================================
"Pan-Germanism was an achievement of intellectuals and writers. The professors of history, law, economics, political science, geography, and philosophy were its most uncompromising advocates. They converted the students of the universities to their ideas. Very soon the graduates made more converts. As teachers in the field of higher education (in the famous German Gymnasium and educational institutions of the same rank), as lawyers, judges, civil servants, and diplomats they had ample opportunity to serve their cause." (http://www.mises.org/etexts/mises/og/chap6.asp)
=========================================================================
The Optimist observes the half-filled cup of wine and shares it with a friend.
The Cynic observes the same half-filled cup and shares the upper half with an abstainer.
The Pessimist sees only the upper half of the cup and demands that
the Cynic share it with the Optimist.
=========================================================================



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From ann.bickers-at-jazi.net Fri Sep 10 14:01:18 2010
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Email: dufresne-at-ms.umanitoba.ca
Name: Andre Dufresne

Organization: University of Manitoba Biology

Title-Subject: [Filtered] Nano Particles

Message: Hi All,
I have a request to image some gold nano rods. I have done this type
of work before with watersoluble particle on carbon coated formvar
grids with success. In this case the particles are hydrophobic. The
PI wants to use chloroform however I'm at a loss as how to image
these particles using such a solvent as a carrier. If anybody has
any suggestions or experience in this area and can point me in the
right direction please let me know. Merci.

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6, 22 -- From zaluzec-at-microscopy.com Sun Sep 12 09:49:40 2010
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From: bozzola-at-siu.edu
Date: Sun, 12 Sep 2010 14:31:53 -0500
Subject: [Microscopy] Re: viaWWW: Nano Particles Imaging

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

Actually, this should be easier than dealing with aqueous solutions.

About the only thing I would change is to use only carbon as a
substrate (since there is a slight possibility that chloroform might
dissolve the Formvar and cause a dirty background). Just take a small
amount of the specimen into a disposable pipette and drop it directly
onto the grid that is resting on a filter paper. Allow to air dry,
then into the TEM. If it is too concentrated, then try a couple
dilutions of the gold in chloroform.

De rien,

JB

On Sun, Sep 12, 2010 at 9:51 AM, {dufresne-at-ms.umanitoba.ca} wrote:
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} Name: Andre Dufresne
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} Organization: University of Manitoba Biology
}
} Title-Subject: [Filtered] Nano Particles
}
} Message: Hi All,
} I have a request to image some gold nano rods.  I have done this type
} of work before with watersoluble particle on carbon coated formvar
} grids with success.  In this case the particles are hydrophobic.  The
} PI wants to use chloroform however I'm at a loss  as how to image
} these particles using such a solvent as a carrier.  If anybody has
} any suggestions or experience in this area and can point me in the
} right direction please let me know.  Merci.
}
}  Login Host: 130.179.112.126
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--
John J. Bozzola, Ph.D., Director
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From: marshall-at-sfu.ca
Date: Sun, 12 Sep 2010 18:23:32 -0500
Subject: [Microscopy] viaWWW: Replacement Si diodes for backscatter electron detectors?

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Email: marshall-at-sfu.ca
Name: Dan Marshall

Organization: Simon Fraser University

Title-Subject: [Filtered] Replacement Si diodes for backscatter
electron detectors?

Message: We recently received a retired GW backscatter electron
detector without the Si detector didoes. We've tested the unit with
some Thor didoes and the unit works, but is not optimal as the diodes
used are not specifically optimized for electron detection. We've
been searching to try and find a vendor for better suited diodes.
Hamamatsu and IRD sell very expensive diodes, but we were hoping that
other labs may have faced similar budgetary constraints and have
found a source of off-the-shelf diodes with more reasonable pricing,
or have some information about what has happened to the GW
electronics Firm.


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From: tomw-at-uidaho.edu
Date: Mon, 13 Sep 2010 08:30:12 -0500
Subject: [Microscopy] viaWWW: nanoparticles in ionic fluid

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Email: tomw-at-uidaho.edu
Name: Tom Williams

Organization: University of Idaho

Title-Subject: [Filtered] nanoparticles in ionic fluid

Message: Greetings all. I have a research group in Chemistry who are
working with gold nanoparticles (approx. 100 nm) carried in an ionic
fluid with a very low vapor pressure. Essentially the fluid doesn't
evaporate even under specimen chamber vacuum (even tried placing
samples in a vacuum evaporator for several hours). Residual fluid on
the stubs makes imaging the particles difficult to impossible (tried
SE, variable pressure, in-lens, BSE-no luck). Would anyone have a
possible procedure or a suggestion?

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From: Woody.White-at-areva.com
Date: Mon, 13 Sep 2010 08:38:01 -0500
Subject: [Microscopy] viaWWW: Replacement Si diodes for backscatter

Contents Retrieved from Microscopy Listserver Archives
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Sometime ago, GW was taken over by KE Developments in the UK. If I
recall correctly, their US rep is EBS (Electron Beam Sciences).

Woody

N.W. (Woody) White Jr.
Senior Electron Microscopist
Chemistry and Materials Center
AREVA NP Inc
An AREVA and Siemens Company
Lynchburg, VA
Lab Phone: 434.832.3004

-----Original Message-----
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Email: marshall-at-sfu.ca
Name: Dan Marshall

Organization: Simon Fraser University

Title-Subject: [Filtered] Replacement Si diodes for backscatter electron
detectors?

Message: We recently received a retired GW backscatter electron detector
without the Si detector didoes. We've tested the unit with some Thor
didoes and the unit works, but is not optimal as the diodes used are not
specifically optimized for electron detection. We've been searching to
try and find a vendor for better suited diodes.
Hamamatsu and IRD sell very expensive diodes, but we were hoping that
other labs may have faced similar budgetary constraints and have found a
source of off-the-shelf diodes with more reasonable pricing, or have
some information about what has happened to the GW electronics Firm.


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From: mdufraine-at-ebsciences.com
Date: Mon, 13 Sep 2010 10:44:11 -0500
Subject: [Microscopy] viaWWW: Replacement Si diodes for backscatter electron detectors?

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

Dan-

Energy Beam Sciences is the US distributor for KE Developments, which bought GW Electronics 6-7 years ago.
If you would contact me off-line I'd be happy to see if we can help, or get you in contact with someone at KE to assist you.

Mike Dufraine
Energy Beam Sciences, Inc.
800-992-9037 X340


-----Original Message-----
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To: Mike Dufraine

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Email: marshall-at-sfu.ca
Name: Dan Marshall

Organization: Simon Fraser University

Title-Subject: [Filtered] Replacement Si diodes for backscatter
electron detectors?

Message: We recently received a retired GW backscatter electron
detector without the Si detector didoes. We've tested the unit with
some Thor didoes and the unit works, but is not optimal as the diodes
used are not specifically optimized for electron detection. We've
been searching to try and find a vendor for better suited diodes.
Hamamatsu and IRD sell very expensive diodes, but we were hoping that
other labs may have faced similar budgetary constraints and have
found a source of off-the-shelf diodes with more reasonable pricing,
or have some information about what has happened to the GW
electronics Firm.


Login Host: 174.7.122.17
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From: bozzola-at-siu.edu
Date: Mon, 13 Sep 2010 11:14:16 -0500
Subject: [Microscopy] Re: viaWWW: nanoparticles in ionic fluid

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

The best way would be, of course, to re-suspend the particles in a
solvent that would evaporate properly (like acetone, chloroform, etc),
leaving behind only the nano materials.

If that is not possible, then try putting the specimen onto a porous
material (like filter paper or a holey, carbon-filmed grid). Allow the
liquid to wick onto the paper (or through the grid), then follow up
with a couple drops of solvent to flush out the problem liquid. Some
nano material should remain behind on the paper surface or trapped in
some of the smaller holes.

Examine in the SEM or TEM (if on a grid). If you have STEM with BSE
detector on your SEM, that might be the best way for the grid.

Let us know how it goes .........

John Bozzola
IMAGE Center
Southern Illinois University
Carbondale, IL 62901

On Mon, Sep 13, 2010 at 8:31 AM, {tomw-at-uidaho.edu} wrote:
}

} Email: tomw-at-uidaho.edu
} Name: Tom Williams
}
} Organization: University of Idaho
}
} Title-Subject: [Filtered] nanoparticles in ionic fluid
}
} Message: Greetings all.  I have a research group in Chemistry who are
} working with gold nanoparticles (approx. 100 nm) carried in an ionic
} fluid with a very low vapor pressure.  Essentially the fluid doesn't
} evaporate even under specimen chamber vacuum (even tried placing
} samples in a vacuum evaporator for several hours).  Residual fluid on
} the stubs makes imaging the particles difficult to impossible (tried
} SE, variable pressure, in-lens, BSE-no luck).  Would anyone have a
} possible procedure or a suggestion?
}


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From: eikonika-at-otenet.gr
Date: Mon, 13 Sep 2010 12:38:55 -0500
Subject: [Microscopy] nanoparticles in ionic fluid

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

Dear Tom
In addition to John's recommendations I would try to place the particles on
to a heat stable surface and then to heat them until completely dried.
yorgos


Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

eikonika-at-otenet.gr
yorgosnikas-at-hotmail.com
Tel/fax +30 210 8957677
mobile +30 6945 107477
*********************************

The best way would be, of course, to re-suspend the particles in a
solvent that would evaporate properly (like acetone, chloroform, etc),
leaving behind only the nano materials.

If that is not possible, then try putting the specimen onto a porous
material (like filter paper or a holey, carbon-filmed grid). Allow the
liquid to wick onto the paper (or through the grid), then follow up
with a couple drops of solvent to flush out the problem liquid. Some
nano material should remain behind on the paper surface or trapped in
some of the smaller holes.

Examine in the SEM or TEM (if on a grid). If you have STEM with BSE
detector on your SEM, that might be the best way for the grid.

Let us know how it goes .........

John Bozzola
IMAGE Center
Southern Illinois University
Carbondale, IL 62901

On Mon, Sep 13, 2010 at 8:31 AM, {tomw-at-uidaho.edu} wrote:
}

} Email: tomw-at-uidaho.edu
} Name: Tom Williams
}
} Organization: University of Idaho
}
} Title-Subject: [Filtered] nanoparticles in ionic fluid
}
} Message: Greetings all. Â I have a research group in Chemistry who are
} working with gold nanoparticles (approx. 100 nm) carried in an ionic
} fluid with a very low vapor pressure. Â Essentially the fluid doesn't
} evaporate even under specimen chamber vacuum (even tried placing
} samples in a vacuum evaporator for several hours). Â Residual fluid on
} the stubs makes imaging the particles difficult to impossible (tried
} SE, variable pressure, in-lens, BSE-no luck). Â Would anyone have a
} possible procedure or a suggestion?
}


__________ Information from ESET NOD32 Antivirus, version of virus signature database 5447 (20100913) __________

The message was checked by ESET NOD32 Antivirus.

http://www.eset.com




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From: connellyps-at-nhlbi.nih.gov
Date: Tue, 14 Sep 2010 13:07:50 -0500
Subject: [Microscopy] Sea Monkey guts

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

Anyone familiar with a Leica TCS NT confocal circa 1997?

A lab close to us has one, but it is not working. Leica no longer
supports this model and I am wondering if it is the trouble to get it
going.

Don't know the exact situation, they say it stopped working after they
shoved (their word) it out of the lab to make room for something else.
I think the operative word here is 'shoved'.

I would kind of like to get this thing at least partially operational.
I talk to my students a lot about confocal scopes but do not have
anything I can actually demonstrate.

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu




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From armando.grottesi-at-cinetech.it Mon Sep 13 19:13:53 2010
Return-Path: {armando.grottesi-at-cinetech.it}
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Received: from [190.41.143.181] by mx1.cinetech.it; Mon, 13 Sep 2010 19:09:44 -0500

Greetings!

A little while back Lene had posted a request for TEM information on
Artemia, ³Sea Monkey², guts that she has been asked to image. She¹d like to
see a few images if someone has taken any since she does not know what they
should look like. Lene did not get any replies.

I suggested that many in the USA were returning from vacation and might have
missed her message. I then volunteered to post this message for her.
She is most interested to learn if others have worked with these tiny
beasts. Please reply to her in Oslo.

Lene Cecilie Hermansen
Elektronmikroskopisk lab, NVH
Tlf; 22 96 45 50
LeneCecilie.Hermansen-at-nvh.no

I was interested in this subject for my uncle, who worked with a large toy
distributer maybe 45 years ago, decided that Sea Monkies would be a big hit
on the toy market. I could not understand how they could be considered a
"toy"!

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E, Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-mail.nih.gov

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and can be freely shared
and reproduced.



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From: oshel1pe-at-cmich.edu
Date: Tue, 14 Sep 2010 13:27:35 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist

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

} Date: Tue, 14 Sep 2010 11:09:07 -0700 (PDT)
} From: "Susan C. Van Horn" {susan.vanhorn-at-sunysb.edu}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Tuesday, September
} 14, 2010 at 11:08:59 AM.
}
} realname - Susan C. Van Horn
} Email - susan.vanhorn-at-sunysb.edu
} ORGANIZATION - SUNY -at- Stony Brook
} EDUCATION - Graduate College
} LOCATION - Stony Brook
} SUBJECT_OF_QUESTION - FEI Tecnai12 BioTwinG2 TEM compu-stage
} QUESTION - We have a FEI Tecnai12 BioTwinG2 TEM with a compu-stage
} that within the past 4 months has been giving us tremndous problems
} with drift and vacuum loss. Was curious and would like to know if
} anyone else is having these problems.
} Thank you,
} Sue

--
***************************************************************************************
Forwarded from "Ask a Microscopist"
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****************************************************************************************

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From: FMonson-at-wcupa.edu
Date: Tue, 14 Sep 2010 15:44:33 -0500
Subject: [Microscopy] RE: Fwd: Ask-A-Microscopist

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

Hi Sue,

We have a Tecnai T12 Twin, but the same stage with the same problems. So, yes the specimen rod port will, periodically, present with the symptoms you describe. When it happens, I call my handy-dandy service engineer, and he comes out, dismantles the stage on its 'rack', puts it back together after cleaning, and then either lucks out with instant vacuum OR has to come back tomorrow to get another mote out of the works.

Good luck,

Fred Monson

Frederick C. Monson, PhD
Technical Director, CMIRT
West Chester University
West Chester, PA, 19383
610-738-0437
New Web Site: http://cmirt.wcupa.edu/index.html

New Scheduler: http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl

Reads of the Month:
1. "The Liver as a Lymphoid Organ," Ian Nicholas Crispe, Annual Review of Immunology, Apr 2009, Vol. 27: 147-163. (Access Depends on subscription.)
2. "A Personal Journey of Discovery: Developing Technology and Changing Biology," Lee Hood, Annual Review of Analytic Chemistry, Vol. 1: 1-43.

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

} Date: Tue, 14 Sep 2010 11:09:07 -0700 (PDT)
} From: "Susan C. Van Horn" {susan.vanhorn-at-sunysb.edu}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Tuesday, September
} 14, 2010 at 11:08:59 AM.
}
} realname - Susan C. Van Horn
} Email - susan.vanhorn-at-sunysb.edu
} ORGANIZATION - SUNY -at- Stony Brook
} EDUCATION - Graduate College
} LOCATION - Stony Brook
} SUBJECT_OF_QUESTION - FEI Tecnai12 BioTwinG2 TEM compu-stage
} QUESTION - We have a FEI Tecnai12 BioTwinG2 TEM with a compu-stage
} that within the past 4 months has been giving us tremndous problems
} with drift and vacuum loss. Was curious and would like to know if
} anyone else is having these problems.
} Thank you,
} Sue

--
***************************************************************************************
Forwarded from "Ask a Microscopist"
Please remember that the person asking the question is likely
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**any reply should go directly to the poster**
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Using the "reply" function in your email does *not* send your answer
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Please copy their email address from their question.
****************************************************************************************

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From: Jerzy.Gazda-at-ceriumlabs.com
Date: Tue, 14 Sep 2010 17:13:25 -0500
Subject: [Microscopy] RE: Fwd: Ask-A-Microscopist

Contents Retrieved from Microscopy Listserver Archives
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We use Cm300 with Compustage, and have similar problems since the tool reached ~10 years of use. Currently our service engineer is claiming that they are having problems with supplier of o-rings and that the last 5 packages all came with defective/substandard materials.

We are working with FEI service organization to resolve that problem. But in the meantime, one thing that works is to tilt few degrees in alpha (X) the specimen rod just after insertion. This somehow seals the o-rings better. Our pump down times can be as long as 30 minutes.

Hope this helps,

Jerzy

-----Original Message-----
X-from: FMonson-at-wcupa.edu [mailto:FMonson-at-wcupa.edu]
Sent: Tuesday, September 14, 2010 3:53 PM
To: Gazda, Jerzy

Hi Sue,

We have a Tecnai T12 Twin, but the same stage with the same problems. So, yes the specimen rod port will, periodically, present with the symptoms you describe. When it happens, I call my handy-dandy service engineer, and he comes out, dismantles the stage on its 'rack', puts it back together after cleaning, and then either lucks out with instant vacuum OR has to come back tomorrow to get another mote out of the works.

Good luck,

Fred Monson

Frederick C. Monson, PhD
Technical Director, CMIRT
West Chester University
West Chester, PA, 19383
610-738-0437
New Web Site: http://cmirt.wcupa.edu/index.html

New Scheduler: http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl

Reads of the Month:
1. "The Liver as a Lymphoid Organ," Ian Nicholas Crispe, Annual Review of Immunology, Apr 2009, Vol. 27: 147-163. (Access Depends on subscription.)
2. "A Personal Journey of Discovery: Developing Technology and Changing Biology," Lee Hood, Annual Review of Analytic Chemistry, Vol. 1: 1-43.

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} Date: Tue, 14 Sep 2010 11:09:07 -0700 (PDT)
} From: "Susan C. Van Horn" {susan.vanhorn-at-sunysb.edu}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Tuesday, September
} 14, 2010 at 11:08:59 AM.
}
} realname - Susan C. Van Horn
} Email - susan.vanhorn-at-sunysb.edu
} ORGANIZATION - SUNY -at- Stony Brook
} EDUCATION - Graduate College
} LOCATION - Stony Brook
} SUBJECT_OF_QUESTION - FEI Tecnai12 BioTwinG2 TEM compu-stage
} QUESTION - We have a FEI Tecnai12 BioTwinG2 TEM with a compu-stage
} that within the past 4 months has been giving us tremndous problems
} with drift and vacuum loss. Was curious and would like to know if
} anyone else is having these problems.
} Thank you,
} Sue

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From: maloneyb-at-fiu.edu
Date: Wed, 15 Sep 2010 10:21:04 -0500
Subject: [Microscopy] Electron Diffraction gallery

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

Have you-all tried ordering your O-rings from the Zatkoff Co
(www.zatkoff.com)?

I've had good luck buying O-rings from them over the past 40 years.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From a.hoppmann-at-g-h-k.de Wed Sep 15 07:22:46 2010
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Dear Group - does anyone know of a published work that provides
diffraction images of elements, compounds, etc.?
Thanks so much.
Barbara



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From: zaluzec-at-aaem.amc.anl.gov
Date: Wed, 15 Sep 2010 15:15:24 -0500
Subject: [Microscopy] Re: Electron Diffraction gallery

Contents Retrieved from Microscopy Listserver Archives
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Dear All,

I am looking for a scan-generator and a scan-amp for a Jeol 35C SEM.
Best would be from somewhere in Europe but I would also pay for parcels coming from overseas :-)

I need the units for repair of a scope I try to bring back to working order at a highschool at Nuernberg / Germany.

Best regards,
Stefan

--
-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

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From abtei-at-koenigsmuenster.de Wed Sep 15 12:35:13 2010
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Received: from [95.179.92.14] by mx0.koenigsmuenster.de; Wed, 15 Sep 2010 20:31:03 +0300

There are 2 old compilations:


Convergent Beam Electron Diffraction of Alloy Phases
pub. Adam Hilger Ltd, Bristol 1984
Edited by J. F. Mansfield

Convergent Beam Electron Diffraction II
pub. by JEOL 1988
by. Tanaka, Terauchi, Kaneyama


Nestor
Your Friendly Neighborhood SysOp



At 10:21 AM -0500 9/15/10, maloneyb-at-fiu.edu wrote:
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--
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Dr. Nestor J. Zaluzec
Argonne National Laboratory
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Tel: 530-NES-TORZ (530-637-8679), Fax: 630-252-4798

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The box said ...
"This program requires Win 95/98/NT or better..."
So I bought a Mac !

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From: glenmac-at-u.washington.edu
Date: Wed, 15 Sep 2010 15:17:26 -0500
Subject: [Microscopy] LED fiber for surgical scope

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

Hello,
One of our colleagues is going overseas on a medical mission to provide ear surgery. He bought a used Zeiss operating microscope that has a fiber bundle for through-lens illumination. Has anyone experience with whether an LED illuminator is sufficiently bright to be used as the light source for this application?

Thanks,
Glen


Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923 USA
(206) 616-4156
glenmac-at-uw.edu










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From: Henrik.Kaker-at-guest.arnes.si
Date: Wed, 15 Sep 2010 15:18:39 -0500
Subject: [Microscopy] Re: Looking for Jeol 35C parts

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On 15.9.2010 18:45, stefan.diller-at-t-online.de wrote:
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} Dear All,
}
} I am looking for a scan-generator and a scan-amp for a Jeol 35C SEM.
} Best would be from somewhere in Europe but I would also pay for parcels coming from overseas :-)
}
} I need the units for repair of a scope I try to bring back to working order at a highschool at Nuernberg / Germany.
}
} Best regards,
} Stefan
}
Stefan,

Please see the link, http://www.kaker.com/used/jeoljsm35cf.html

Best regards,

Henrik

--
Dr. Henrik Kaker
Ob Suhi 23
SI-2390 Ravne
Slovenia
GSM: +386 31 784 281
http://www.kaker.com
Email:info-at-kaker.com


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From: wleblan-at-lsu.edu
Date: Wed, 15 Sep 2010 19:12:53 -0500
Subject: [Microscopy] viaWWW: Solid State Detectors for XRD

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Email: wleblan-at-lsu.edu
Name: Wanda S LeBlanc

Organization: LSU, Geology and Geophysics Department

Title-Subject: [Filtered] Solid State Detectors for XRD

Message: Dear Group,

I am looking for a replacement for our Kevex Psi Peltier Cooled
Silicon Detector for our Bruker/Siemens D5000 XRD. Does anyone know
where I can get a replacement or get this detector repaired. Kevex
sent me to this group.

Thanks,

Wanda

Login Host: 130.39.47.155
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==============================Original Headers==============================
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From: ropope-at-gmail.com
Date: Wed, 15 Sep 2010 19:46:31 -0500
Subject: [Microscopy] viaWWW: GSR course

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Email: ropope-at-gmail.com
Name: Robert Pope

Title-Subject: [Filtered] GSR course

Message: Good day everyone. I continue to enjoy reading all the
posts, and sporadically manage to develop my own questions. We are
interested in adding to our basic expertise, and was wondering how
many courses there are for GSR/particle analysis in the US.
Especially if the facility is ISO accredited, or perhaps knows what
ISO entails. Our facility will eventually be ISO accredited, and we
will want to add the GSR or particle analysis to our list of
potential offerings. I know that there is one course around Chicago,
but am looking for one a little closer to the DC area. Although
anywhere in the US is acceptable. All information is welcome.
Thank you,
Robert

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==============================Original Headers==============================
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From: colijn.1-at-osu.edu
Date: Wed, 15 Sep 2010 19:49:31 -0500
Subject: [Microscopy] Re: viaWWW: Solid State Detectors for XRD

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

Hi Wanda,

Kevex was absorbed by Noran which was then absorbed by Thermo. I would
be surprised if you can get any kind of manufacturer service on your
detector today.

There may be some 3rd party service organizations that can work on these
detectors. You can try Doug at Tracor Northern Analyzer Service
(www.tnas.net) or Advanced Analysis Technologies
(http://www.advancedanalysistech.com/services.html). I'm sure there are
more EDX service companies, perhaps they will contact you. You may,
however, be better off in going with a new detector.

I believe that Moxtek (www.moxtek.com) makes energy-dispersive detectors
for the XRD market. It is possible that there is "some assembly
required". If you talk to them, they should be able to point you in the
right direction.

Cheers,
Henk
(no financial or other interest in Moxtek or any other company)



At 9/15/2010 8:14 PM, wleblan-at-lsu.edu wrote:
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}
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}
} Email: wleblan-at-lsu.edu
} Name: Wanda S LeBlanc
}
} Organization: LSU, Geology and Geophysics Department
}
} Title-Subject: [Filtered] Solid State Detectors for XRD
}
} Message: Dear Group,
}
} I am looking for a replacement for our Kevex Psi Peltier Cooled
} Silicon Detector for our Bruker/Siemens D5000 XRD. Does anyone know
} where I can get a replacement or get this detector repaired. Kevex
} sent me to this group.
}
} Thanks,
}
} Wanda
}
} Login Host: 130.39.47.155
} ---------------------------------------------------------------------------
}
} ==============================Original Headers==============================
} 9, 22 -- From zaluzec-at-microscopy.com Wed Sep 15 19:12:53 2010
} 9, 22 -- Received: from znl.com ([206.69.208.20])
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} 9, 22 -- Subject: viaWWW: Solid State Detectors for XRD
} 9, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed"
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}

--
Hendrik O. Colijn
www.ceof.ohio-state.edu
OSU Campus Electron Optics Facility colijn.1-at-osu.edu
040 Fontana Labs (614) 292-0674 (V)
116 W. 19th Ave. (614) 292-7523 (F)
Columbus, OH 43210

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

==============================Original Headers==============================
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From: jteshima-at-dunesciences.com
Date: Wed, 15 Sep 2010 20:10:38 -0500
Subject: [Microscopy] viaWWW: nanoparticles in ionic fluid

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

The recent postings on List server indicate a growing interest in
simple, reproducible methods to prepare nanoparticles for metrology
and chemical characterization. We have found that by functionalizing
the surface of the TEM grid through surface charge and chemistry TEM
preparation is significantly improved with respect to capture and
dispersion. There are some interesting TEM images, papers and
training videos on our website with specific examples for gold and
silver.

http://www.dunesciences.com/prep_video.html
http://www.dunesciences.com/nanogrids.html

Janet

Janet Teshima
SMART Grids Product Manager
cell: 503-5447526
office: 541-636-3712
jteshima-at-dunesciences.com
www.dunesciences.com

On Sep 13, 2010, at 9:21 AM, bozzola-at-siu.edu wrote:

}
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}
} The best way would be, of course, to re-suspend the particles in a
} solvent that would evaporate properly (like acetone, chloroform, etc),
} leaving behind only the nano materials.
}
} If that is not possible, then try putting the specimen onto a porous
} material (like filter paper or a holey, carbon-filmed grid). Allow the
} liquid to wick onto the paper (or through the grid), then follow up
} with a couple drops of solvent to flush out the problem liquid. Some
} nano material should remain behind on the paper surface or trapped in
} some of the smaller holes.
}
} Examine in the SEM or TEM (if on a grid). If you have STEM with BSE
} detector on your SEM, that might be the best way for the grid.
}
} Let us know how it goes .........
}
} John Bozzola
} IMAGE Center
} Southern Illinois University
} Carbondale, IL 62901
}
} On Mon, Sep 13, 2010 at 8:31 AM, {tomw-at-uidaho.edu} wrote:
} }
}
} } Email: tomw-at-uidaho.edu
} } Name: Tom Williams
} }
} } Organization: University of Idaho
} }
} } Title-Subject: [Filtered] nanoparticles in ionic fluid
} }
} } Message: Greetings all. I have a research group in Chemistry who are
} } working with gold nanoparticles (approx. 100 nm) carried in an ionic
} } fluid with a very low vapor pressure. Essentially the fluid doesn't
} } evaporate even under specimen chamber vacuum (even tried placing
} } samples in a vacuum evaporator for several hours). Residual fluid on
} } the stubs makes imaging the particles difficult to impossible (tried
} } SE, variable pressure, in-lens, BSE-no luck). Would anyone have a
} } possible procedure or a suggestion?
} }
}
}
} ==============================Original
} Headers==============================
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} 8, 20 -- Subject: Re: [Microscopy] viaWWW: nanoparticles in ionic
} fluid
} 8, 20 -- From: John Bozzola {bozzola-at-siu.edu}
} 8, 20 -- To: MSAListserver {Microscopy-at-microscopy.com} ,
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From: sarj0007-at-unf.edu
Date: Fri, 17 Sep 2010 09:23:49 -0500
Subject: [Microscopy] Thin section thickness guides and sorvall MT2

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Email: tomw-at-uidaho.edu
Name: Thomas Williams

Organization: UI

Title-Subject: [Filtered] Ionic Liquids-Many Thanks!

Message: Good morning, good afternoon, or good evening. I wanted to
take a moment to thank all those who sent along the great advice on
ionic liquids and nanoparticles. Enlightening. Now, I will send the
info to the Grad student and make him do all the work!

Cheers,
Tom


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From apiechotta-at-thummahr.de Fri Sep 17 03:38:46 2010
Return-Path: {apiechotta-at-thummahr.de}
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Received: from [61.213.216.65] by obiwan.thummahr.de; Fri, 17 Sep 2010 17:34:33 +0900

Good day all. I have two questions. First I am looking for a good color chart for measuring the quality of a thin section. My professor describes these lovely full color laminated items that she has seen in most of the EM centers with an ultra-microtome has on a wall, and a lot of companies used to give them away at conferences(as she describes it from many years past). Does anyone know a good source for finding these? I haven't had much look searching for them online and she just wants an image for us to be able to look up and quickly tell where we are in thickness. I have it in a text chart out of Peace's 1964 book.

Secondly, our Sorvall MT2 has been giving us problems. Sometimes it would stop moving when we change the speed setting, and we would have to go down or up in speed until it would start to move again. Last weekend I pulled it apart and think I've found the culprit. There are two large discs on the right hand side of the machine, inside. The outer disc is connected to the motor and the hand crank. The inner disc turns the gears for the rest of the machine. In between is a little wheel perpendicular to the two large wheels such that when one disc turns, it turns the small wheel forcing the other disc to turn. I think the either the grip on the rubber of the small wheel has worn down over the years, or maybe something was supposed to be coated on the larger wheels has worn away. Has anyone had this problem with a fix or a good source of parts for this microtome? I have it working reliably right now by pushing the outer disc closer but its too close to turn the speed screw so we are stuck at only one speed setting and cannot adjust without taking it apart, moving the disc, moving the screw, moving the disc closer again, and closing it up. I think most people would agree that is not optimal when starting to get good sections :)

Then again stopping in the middle of getting good sections hasnt been a problem yet, and I find it unsettling that I wish to have a particular problem, since that would mean I'm getting good sections. Ha. Thanks.


-Jason Saredy


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From: oshel1pe-at-cmich.edu
Date: Fri, 17 Sep 2010 09:39:14 -0500
Subject: [Microscopy] Re: Thin section thickness guides and sorvall MT2

Contents Retrieved from Microscopy Listserver Archives
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Jason,

The little wheel in between the big wheels (crank wheel & "drive"
wheel) is the speed regular - or CVT. It has an o-ring running around
the circumference. Replace this o-ring with any high quality o-ring
(like Viton). Try hard to get an o-ring with the same cross-sectional
size; or, put another way, with the same inner diameter and outer
diameter. A little variance here isn't too evil, but it means the
speed settings will be different from what you're used to.

If you don't find a local source, I have an extra section thickness
color chart or two I can send you - let me know.

Phil
P.S. It's also in Norma Reid's excellent book "Ultramicrotomy", in
the Prac. Meth. EM series edited by Audrey Glauert.

} ----------------------------------------------------------------------------
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Biology Department
024C Brooks Hall
Central Michigan University
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(989) 774-3576

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From: stefan.diller-at-t-online.de
Date: Fri, 17 Sep 2010 10:04:59 -0500
Subject: [Microscopy] Looking for Jeol 35C parts - Thanks

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

Jason,

I am still using an MT2 that was being discarded so it needed a bit of
reconditioning which was quite easy having used one for a few decades
previously. I will contact you off the Listserv. This is not a big problem
once you know how to fix it. Again, the pre-computer machines are user
friendly. In the mean time do not put anything on the two metal wheels!

If anyone else wishes the information contact me and I¹ll send it to you at
the same time.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E ­ Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
connellyps-at-mail.nih.gov

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and can be freely shared
and reproduced.
===

X-from: {sarj0007-at-unf.edu}
Reply-To: {sarj0007-at-unf.edu}

Dear All,
thank you for your kind help with parts for the Jeol 35C.
I had been able to fix the problem yesterday.
I appreciate the help of all on this list very much :-)

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------


}
} Dear All,
}
} I am looking for a scan-generator and a scan-amp for a Jeol 35C SEM.
} Best would be from somewhere in Europe but I would also pay for parcels coming from overseas :-)
}
} I need the units for repair of a scope I try to bring back to working order at a highschool at Nuernberg / Germany.
}
} Best regards,
} Stefan
}

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From: bozzola-at-siu.edu
Date: Fri, 17 Sep 2010 10:41:22 -0500
Subject: [Microscopy] Re: Thin section thickness guides and sorvall MT2

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

Regarding the laminated, section-thickness, color scale distributed by
Sorvall, often given away with their ultramicrotomes. This was based
on the work of Lee Peachey.

I don't believe the original is copyrighted (but check beforehand), so
why not borrow the original from your professor and scan it using a
high quality, color scanner?

John Bozzola
IMAGE Center
Southern Illinois University
Carbondale, IL

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From: contact-at-integrityscientific.com
Date: Fri, 17 Sep 2010 10:46:24 -0500
Subject: [Microscopy] Specimen prep - precision mechanical lapping/polishing

Contents Retrieved from Microscopy Listserver Archives
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Dear Listers,
I am looking for a high precision lapping/polishing machine for TEM
preparation (e.g. wedge polishing etc). I know of the Allied Hi-Tech
system - can anyone tell me if there any other similar machines which I
should be considering? Manufacturers are welcome to respond off-list.

Best Regards

Richard

==============================Original Headers==============================
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From: bozzola-at-siu.edu
Date: Fri, 17 Sep 2010 12:41:02 -0500
Subject: [Microscopy] Thin section thickness guides and sorvall MT2

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

Wouldn't it be great to see something like this in Microscopy Today?
Hint, hint, hint .....

--

John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


On Fri, Sep 17, 2010 at 11:16 AM, Factor, Jan {Jan.Factor-at-purchase.edu} wrote:
} If John Bozzola is correct, and the color chart is not under copyright, then
} perhaps someone who has a copy that is still in good condition would be
} willing to scan it and make it available to the list. Just a thought...
} --Best, Jan Factor
}
} Jan Robert Factor, Ph.D.
} Professor of Biology
} Purchase College, State University of New York
} Purchase, NY 10577
} Office: 2016NS, 914-251-6659
} Office Hours (Fall 2010): Mon., 10:30-12:00, Wed., 4:30-6:00
} jan.factor-at-purchase.edu
}
} Please consider the environment before printing this e-mail
} ________________________________________
} From: bozzola-at-siu.edu [bozzola-at-siu.edu]
} Sent: Friday, September 17, 2010 11:52 AM
} To: Factor, Jan
} Subject: [Microscopy] Re: Thin section thickness guides and sorvall MT2
}
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
}
} Regarding the laminated, section-thickness, color scale distributed by
} Sorvall, often given away with their ultramicrotomes. This was based
} on the work of Lee Peachey.
}
} I don't believe the original is copyrighted (but check beforehand), so
} why not borrow the original from your professor and scan it using a
} high quality, color scanner?
}
} John Bozzola
} IMAGE Center
} Southern Illinois University
} Carbondale, IL
}
} ==============================Original Headers==============================
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} 3, 18 -- Date: Fri, 17 Sep 2010 10:41:18 -0500
} 3, 18 -- Message-ID:
} {AANLkTikHJrOLa=pF1xzdJm26gKbGm0C1BpV2UC_8NnnR-at-mail.gmail.com}
} 3, 18 -- Subject: Re: [Microscopy] Thin section thickness guides and sorvall
} MT2
} 3, 18 -- From: John Bozzola {bozzola-at-siu.edu}
} 3, 18 -- To: MSAListserver {Microscopy-at-microscopy.com}
} 3, 18 -- Content-Type: text/plain; charset=UTF-8
} ==============================End of - Headers==============================


==============================Original Headers==============================
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6, 21 -- {A395C4E6EB90404ABBC720ACFA8CF130067446E982-at-svexchmb01.purchase.edu}
6, 21 -- Date: Fri, 17 Sep 2010 12:41:00 -0500
6, 21 -- Message-ID: {AANLkTi=7Q-beAKZsDzN1kCzBaApmyy7GwvOtAef502Nk-at-mail.gmail.com}
6, 21 -- Subject: Re: [Microscopy] Re: Thin section thickness guides and sorvall MT2
6, 21 -- From: John Bozzola {bozzola-at-siu.edu}
6, 21 -- To: MSAListserver {Microscopy-at-microscopy.com}
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From: bozzola-at-siu.edu
Date: Fri, 17 Sep 2010 13:02:19 -0500
Subject: [Microscopy] Re: Thin section thickness guides and sorvall MT2

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

Here is a link to Dr. Lee Peachey's original article. Though no color
chart, the paper is a classic.

jcb.rupress.org/content/4/3/233.full.pdf

John Bozzola

On Fri, Sep 17, 2010 at 12:42 PM, {bozzola-at-siu.edu} wrote:
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} Wouldn't it be great to see something like this in Microscopy Today?
} Hint, hint, hint .....
}
} --
}
} John J. Bozzola, Ph.D., Director
} I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
} Southern Illinois University
} 750 Communications Drive
} Carbondale, IL  62901
} Phone: 618-453-3730
}
}
} On Fri, Sep 17, 2010 at 11:16 AM, Factor, Jan {Jan.Factor-at-purchase.edu} wrote:
} } If John Bozzola is correct, and the color chart is not under copyright, then
} } perhaps someone who has a copy that is still in good condition would be
} } willing to scan it and make it available to the list. Just a thought...
} } --Best, Jan Factor
} }
} } Jan Robert Factor, Ph.D.
} } Professor of Biology
} } Purchase College, State University of New York
} } Purchase, NY 10577
} } Office: 2016NS, 914-251-6659
} } Office Hours (Fall 2010): Mon., 10:30-12:00, Wed., 4:30-6:00
} } jan.factor-at-purchase.edu
} }
} } Please consider the environment before printing this e-mail
} } ________________________________________
} } From: bozzola-at-siu.edu [bozzola-at-siu.edu]
} } Sent: Friday, September 17, 2010 11:52 AM
} } To: Factor, Jan
} } Subject: [Microscopy] Re: Thin section thickness guides and sorvall MT2
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} } To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } Regarding the laminated, section-thickness, color scale distributed by
} } Sorvall, often given away with their ultramicrotomes. This was based
} } on the work of Lee Peachey.
} }
} } I don't believe the original is copyrighted (but check beforehand), so
} } why not borrow the original from your professor and scan it using a
} } high quality, color scanner?
} }
} } John Bozzola
} } IMAGE Center
} } Southern Illinois University
} } Carbondale, IL
} }
} } ==============================Original Headers==============================
} } 3, 18 -- From bozzola-at-siu.edu Fri Sep 17 10:41:21 2010
} } 3, 18 -- Received: from mail-ww0-f49.google.com (mail-ww0-f49.google.com
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} } 3, 18 -- References: {201009171424.o8HEOvn0027351-at-ns.microscopy.com}
} } 3, 18 -- Date: Fri, 17 Sep 2010 10:41:18 -0500
} } 3, 18 -- Message-ID:
} } {AANLkTikHJrOLa=pF1xzdJm26gKbGm0C1BpV2UC_8NnnR-at-mail.gmail.com}
} } 3, 18 -- Subject: Re: [Microscopy] Thin section thickness guides and sorvall
} } MT2
} } 3, 18 -- From: John Bozzola {bozzola-at-siu.edu}
} } 3, 18 -- To: MSAListserver {Microscopy-at-microscopy.com}
} } 3, 18 -- Content-Type: text/plain; charset=UTF-8
} } ==============================End of - Headers==============================
}
}
} ==============================Original Headers==============================
} 6, 21 -- From bozzola-at-siu.edu Fri Sep 17 12:41:01 2010
} 6, 21 -- Received: from mail-wy0-f169.google.com (mail-wy0-f169.google.com [74.125.82.169])
} 6, 21 --        by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id o8HHf1LT030248
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} 6, 21 --  Sep 2010 10:41:00 -0700 (PDT)
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} 6, 21 -- References: {201009171552.o8HFqC8j015808-at-ns.microscopy.com}
} 6, 21 --         {A395C4E6EB90404ABBC720ACFA8CF130067446E982-at-svexchmb01.purchase.edu}
} 6, 21 -- Date: Fri, 17 Sep 2010 12:41:00 -0500
} 6, 21 -- Message-ID: {AANLkTi=7Q-beAKZsDzN1kCzBaApmyy7GwvOtAef502Nk-at-mail.gmail.com}
} 6, 21 -- Subject: Re: [Microscopy] Re: Thin section thickness guides and sorvall MT2
} 6, 21 -- From: John Bozzola {bozzola-at-siu.edu}
} 6, 21 -- To: MSAListserver {Microscopy-at-microscopy.com}
} 6, 21 -- Content-Type: text/plain; charset=UTF-8
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}



--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


==============================Original Headers==============================
8, 20 -- From bozzola-at-siu.edu Fri Sep 17 13:02:19 2010
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8, 20 -- Date: Fri, 17 Sep 2010 13:02:18 -0500
8, 20 -- Message-ID: {AANLkTim8QrCtz690BpaqTNUz1ZGFqZ6fGCd6ZUCP5YDr-at-mail.gmail.com}
8, 20 -- Subject: Re: [Microscopy] Thin section thickness guides and sorvall MT2
8, 20 -- From: John Bozzola {bozzola-at-siu.edu}
8, 20 -- To: MSAListserver {Microscopy-at-microscopy.com}
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From: biology-at-ucla.edu
Date: Fri, 17 Sep 2010 13:17:22 -0500
Subject: [Microscopy] Thin section thickness guides and sorvall MT2

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

Well, I have one from Harris Diamond Corportaion Ultramicrotome
Section Color Reference chart, but have no way to scan it currently.
(No scanner nearby)

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Here is a link to Dr. Lee Peachey's original article. Though no color
} chart, the paper is a classic.
}
} jcb.rupress.org/content/4/3/233.full.pdf
}
} John Bozzola
}
} On Fri, Sep 17, 2010 at 12:42 PM, {bozzola-at-siu.edu} wrote:
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor:  The Microscopy Society
} } of America
} } To  Subscribe/Unsubscribe --
} } http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } Wouldn't it be great to see something like this in Microscopy Today?
} } Hint, hint, hint .....
} }
} } --
} }
} } John J. Bozzola, Ph.D., Director
} } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
} } Southern Illinois University
} } 750 Communications Drive
} } Carbondale, IL  62901
} } Phone: 618-453-3730
} }
} }
} } On Fri, Sep 17, 2010 at 11:16 AM, Factor, Jan
} } {Jan.Factor-at-purchase.edu} wrote:
} } } If John Bozzola is correct, and the color chart is not under
} } } copyright, then
} } } perhaps someone who has a copy that is still in good condition would be
} } } willing to scan it and make it available to the list. Just a thought...
} } } --Best, Jan Factor
} } }
} } } Jan Robert Factor, Ph.D.
} } } Professor of Biology
} } } Purchase College, State University of New York
} } } Purchase, NY 10577
} } } Office: 2016NS, 914-251-6659
} } } Office Hours (Fall 2010): Mon., 10:30-12:00, Wed., 4:30-6:00
} } } jan.factor-at-purchase.edu
} } }
} } } Please consider the environment before printing this e-mail
} } } ________________________________________
} } } From: bozzola-at-siu.edu [bozzola-at-siu.edu]
} } } Sent: Friday, September 17, 2010 11:52 AM
} } } To: Factor, Jan
} } } Subject: [Microscopy] Re: Thin section thickness guides and sorvall MT2
} } }
} } } ----------------------------------------------------------------------------
} } } The Microscopy ListServer -- CoSponsor:  The Microscopy Society
} } } of America
} } } To  Subscribe/Unsubscribe --
} } } http://www.microscopy.com/MicroscopyListserver
} } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } } ----------------------------------------------------------------------------
} } }
} } } Regarding the laminated, section-thickness, color scale distributed by
} } } Sorvall, often given away with their ultramicrotomes. This was based
} } } on the work of Lee Peachey.
} } }
} } } I don't believe the original is copyrighted (but check beforehand), so
} } } why not borrow the original from your professor and scan it using a
} } } high quality, color scanner?
} } }
} } } John Bozzola
} } } IMAGE Center
} } } Southern Illinois University
} } } Carbondale, IL
} } }
} } } ==============================Original
} } } Headers==============================
} } } 3, 18 -- From bozzola-at-siu.edu Fri Sep 17 10:41:21 2010
} } } 3, 18 -- Received: from mail-ww0-f49.google.com (mail-ww0-f49.google.com
} } } [74.125.82.49])
} } } 3, 18 --        by ns.microscopy.com
} } } (8.12.11.20060308/8.12.8) with ESMTP id
} } } o8HFfL2x024731
} } } 3, 18 --        for
} } } {Microscopy-at-microscopy.com} ; Fri, 17 Sep 2010 10:41:21
} } } -0500
} } } 3, 18 -- Received: by wwb24 with SMTP id 24so2620706wwb.18
} } } 3, 18 --         for
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} } } -0700 (PDT)
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} } } 3, 18 --  17 Sep 2010 08:41:18 -0700 (PDT)
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} } } -0700 (PDT)
} } } 3, 18 -- In-Reply-To: {201009171424.o8HEOvn0027351-at-ns.microscopy.com}
} } } 3, 18 -- References: {201009171424.o8HEOvn0027351-at-ns.microscopy.com}
} } } 3, 18 -- Date: Fri, 17 Sep 2010 10:41:18 -0500
} } } 3, 18 -- Message-ID:
} } } {AANLkTikHJrOLa=pF1xzdJm26gKbGm0C1BpV2UC_8NnnR-at-mail.gmail.com}
} } } 3, 18 -- Subject: Re: [Microscopy] Thin section thickness guides
} } } and sorvall
} } } MT2
} } } 3, 18 -- From: John Bozzola {bozzola-at-siu.edu}
} } } 3, 18 -- To: MSAListserver {Microscopy-at-microscopy.com}
} } } 3, 18 -- Content-Type: text/plain; charset=UTF-8
} } } ==============================End of -
} } } Headers==============================
} }
} }
} } ==============================Original Headers==============================
} } 6, 21 -- From bozzola-at-siu.edu Fri Sep 17 12:41:01 2010
} } 6, 21 -- Received: from mail-wy0-f169.google.com
} } (mail-wy0-f169.google.com [74.125.82.169])
} } 6, 21 --        by ns.microscopy.com
} } (8.12.11.20060308/8.12.8) with ESMTP id o8HHf1LT030248
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} } 17 Sep 2010 12:41:01 -0500
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} } 6, 21 -- In-Reply-To:
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} } 6, 21 -- References: {201009171552.o8HFqC8j015808-at-ns.microscopy.com}
} } 6, 21 --      
} }   {A395C4E6EB90404ABBC720ACFA8CF130067446E982-at-svexchmb01.purchase.edu}
} } 6, 21 -- Date: Fri, 17 Sep 2010 12:41:00 -0500
} } 6, 21 -- Message-ID:
} } {AANLkTi=7Q-beAKZsDzN1kCzBaApmyy7GwvOtAef502Nk-at-mail.gmail.com}
} } 6, 21 -- Subject: Re: [Microscopy] Re: Thin section thickness
} } guides and sorvall MT2
} } 6, 21 -- From: John Bozzola {bozzola-at-siu.edu}
} } 6, 21 -- To: MSAListserver {Microscopy-at-microscopy.com}
} } 6, 21 -- Content-Type: text/plain; charset=UTF-8
} } 6, 21 -- Content-Transfer-Encoding: 8bit
} } 6, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by
} } ns.microscopy.com id o8HHf1LT030248
} } ==============================End of - Headers==============================
} }
}
}
}
} --
} John J. Bozzola, Ph.D., Director
} I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
} Southern Illinois University
} 750 Communications Drive
} Carbondale, IL  62901
} Phone: 618-453-3730
}
}
} ==============================Original Headers==============================
} 8, 20 -- From bozzola-at-siu.edu Fri Sep 17 13:02:19 2010
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} (mail-ww0-f49.google.com [74.125.82.49])
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} 8, 20 -- Date: Fri, 17 Sep 2010 13:02:18 -0500
} 8, 20 -- Message-ID:
} {AANLkTim8QrCtz690BpaqTNUz1ZGFqZ6fGCd6ZUCP5YDr-at-mail.gmail.com}
} 8, 20 -- Subject: Re: [Microscopy] Thin section thickness guides and
} sorvall MT2
} 8, 20 -- From: John Bozzola {bozzola-at-siu.edu}
} 8, 20 -- To: MSAListserver {Microscopy-at-microscopy.com}
} 8, 20 -- Content-Type: text/plain; charset=UTF-8
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==============================Original Headers==============================
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From: bozzola-at-siu.edu
Date: Fri, 17 Sep 2010 13:25:20 -0500
Subject: [Microscopy] Thin section thickness guides and sorvall MT2

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

Everything You Ever Wanted to Know About Section Thickness and
Interference Colors ....

I just examined two original "Continuous Interference Color and
Thickness Scale for Thin Sections" charts that we have. One is from
Sorvall, the other from DuPont. Neither show a copyright mark, so it
may be OK to scan them. However, if anyone knows otherwise, let me
know. Possibly, someone from RMC Products (Boeckeler Instruments)
might know about this.

If I recall correctly, some microscopy vendors used to supply these
charts, often free of charge.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


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From: PhillipsT-at-missouri.edu
Date: Fri, 17 Sep 2010 13:55:24 -0500
Subject: [Microscopy] Thin section thickness guides and sorvall MT2

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

Check out Lee Peachy's original 1958 article on using interference colors to judge section thickness. A Study of Section Thickness and Physical Distortion Produced during Microtomy http://jcb.rupress.org/content/4/3/233.full.pdf Unfortunately no plate with the color guide.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: bozzola-at-siu.edu [mailto:bozzola-at-siu.edu]
Sent: Friday, September 17, 2010 1:26 PM
To: Phillips, Thomas E.

Everything You Ever Wanted to Know About Section Thickness and
Interference Colors ....

I just examined two original "Continuous Interference Color and
Thickness Scale for Thin Sections" charts that we have. One is from
Sorvall, the other from DuPont. Neither show a copyright mark, so it
may be OK to scan them. However, if anyone knows otherwise, let me
know. Possibly, someone from RMC Products (Boeckeler Instruments)
might know about this.

If I recall correctly, some microscopy vendors used to supply these
charts, often free of charge.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


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From: rschmitz-at-uwsp.edu
Date: Fri, 17 Sep 2010 14:21:07 -0500
Subject: [Microscopy] RE: Thin section thickness guides and sorvall MT2

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


Our lab purchase two UC6 microtomes from Leica a few years ago and a triangular shaped card showing the interference colors came with each machine. Last year when the Leica rep came to repair one of the machines he left a third card behind. Perhaps it is possible to get some of these reference cards from Leica


Bob
Dr. Robert J. Schmitz
Associate Professor of Anatomical Sciences
Department of Biology
University of Wisconsin-Stevens Point
800 Reserve Street, 380 TNR Bld
Stevens Point, WI 54481
715-346-2420
Email: rschmitz-at-uwsp.edu
http://www.uwsp.edu/biology/faculty/rschmitz/index.html



________________________________



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From: mmashore-at-vapop.ucsd.edu
Date: Fri, 17 Sep 2010 14:46:48 -0500
Subject: [Microscopy] viaWWW: Air Bubbles in My blocks

Contents Retrieved from Microscopy Listserver Archives
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This Question/Comment was submitted to the Microscopy Listserver
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Email: mmashore-at-vapop.ucsd.edu
Name: Michael Mashore

Organization: Veterans Medical Research Foundation

Title-Subject: [Filtered] Air Bubbles in My blocks

Message: Hello,

The last couple of specimens I have embedded have
had air bubbles towards the base of my blocks.
The air bubbles are only towards the base were I
put a label (printed by a laserjet printer), and
are not near my tissue.

I am using flat molds that are placed in a glass
vacuum desiccator overnight. The bubbles do not
appear after I remove the samples from the
desiccator. However after polymerizing at 60ƒC
under vacuum for 24h they show up.

Here is the dehydration and embedding protocol:
1. 70% ETOH 10 min.
2. 80% ETOH 10 min.
3. 95% ETOH 10 min.
4. 100% ETOH 10 min.
5. 100% ETOH 10 min.
6. Propylene Oxide + 100% ETOH 1:1 10 min.
7. Propylene Oxide 10 min.
8. Propylene Oxide 5 min.

9. Propylene Oxide + Embed 812 3:1 (rotated) 1 hr.
10. Propylene Oxide + Embed 812 1:1 (rotated) overnight
11. Propylene Oxide + Embed 812 1:3 (rotated) 3 hr.
12. Embed 812 (rotated) overnight
13. Fresh Embed 812 in molds in desiccator overnight
14. 60ƒC vacuum oven 24 hr.

Does anyone have any suggestions?

Thanks,

Michael

Login Host: 132.239.85.200
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From: jkrupp-at-deltacollege.edu
Date: Fri, 17 Sep 2010 17:27:21 -0500
Subject: [Microscopy] ETEC filaments?

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

Hi

Some of you old timers may remember the venerable ETEC SEM. We have
one we use for training students hands on for filament exchange etc.
The scope no longer works, but we keep the column so students can
practice.

Part of their training involves cleaning the gun parts and
reassembling the filament, grid cap, etc. We had a stash of old
filaments they used to practice centering, but today two of our supply
were broken. This exhausts our supply so we are asking if anyone has
some old ETEC filaments in a drawer that you could send our way, or
ideas about substitutes we could use for this exercise.

Thanks

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu




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From: kenconverse-at-qualityimages.biz
Date: Fri, 17 Sep 2010 17:41:49 -0500
Subject: [Microscopy] ETEC filaments?

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

Jon,
I don't really like to think of myself as an "oldtimer", but maybe that's
just vanity.

First, why isn't your ETEC Autoscan still running? It should be. It's what
I consider to be the most student-proof SEM available.

Second, I believe Gene Taylor still has ETEC filaments available.
http://www.semicro.org/search.aspx?find=etec+filaments

I'm glad he does because I have ETEC customers still running their systems
on a daily basis. Why isn't yours running?

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: jkrupp-at-deltacollege.edu [mailto:jkrupp-at-deltacollege.edu]
Sent: Friday, September 17, 2010 6:31 PM
To: kenconverse-at-qualityimages.biz

Hi

Some of you old timers may remember the venerable ETEC SEM. We have
one we use for training students hands on for filament exchange etc.
The scope no longer works, but we keep the column so students can
practice.

Part of their training involves cleaning the gun parts and
reassembling the filament, grid cap, etc. We had a stash of old
filaments they used to practice centering, but today two of our supply
were broken. This exhausts our supply so we are asking if anyone has
some old ETEC filaments in a drawer that you could send our way, or
ideas about substitutes we could use for this exercise.

Thanks

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu




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24, 28 -- From kenconverse-at-qualityimages.biz Fri Sep 17 17:41:48 2010
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From: John.Mardinly-at-wdc.com
Date: Fri, 17 Sep 2010 20:57:54 -0500
Subject: [Microscopy] ETEC filaments?

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




Ken;
When I was at Lockheed in the early 1980's (oh, I guess that makes
me an old-timer) our ETEC was a favorite tool, except for the little problem
that the ssecondary electron detector high voltage bias supply failed every
couple of months, which was not a big deal since the manufacturing and
service was local, and we got a replacement supply the next day. As soon as
ETEC was bought out by Perkin Elmer and service and support were dropped, we
had to replace it with a different brand SEM.

John Mardinly
Western Digital

-----Original Message-----
X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz]
Sent: Friday, September 17, 2010 3:49 PM
To: John Mardinly

Jon,
I don't really like to think of myself as an "oldtimer", but maybe that's
just vanity.

First, why isn't your ETEC Autoscan still running? It should be. It's what
I consider to be the most student-proof SEM available.

Second, I believe Gene Taylor still has ETEC filaments available.
http://www.semicro.org/search.aspx?find=etec+filaments

I'm glad he does because I have ETEC customers still running their systems
on a daily basis. Why isn't yours running?

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: jkrupp-at-deltacollege.edu [mailto:jkrupp-at-deltacollege.edu]
Sent: Friday, September 17, 2010 6:31 PM
To: kenconverse-at-qualityimages.biz

Hi

Some of you old timers may remember the venerable ETEC SEM. We have
one we use for training students hands on for filament exchange etc.
The scope no longer works, but we keep the column so students can
practice.

Part of their training involves cleaning the gun parts and
reassembling the filament, grid cap, etc. We had a stash of old
filaments they used to practice centering, but today two of our supply
were broken. This exhausts our supply so we are asking if anyone has
some old ETEC filaments in a drawer that you could send our way, or
ideas about substitutes we could use for this exercise.

Thanks

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu




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From: colijn.1-at-osu.edu
Date: Fri, 17 Sep 2010 21:42:00 -0500
Subject: [Microscopy] Re: ETEC filaments?

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

Hi Jonathan,

In the "old days" we used to send our burned out filament bases to
Energy Beam Sciences to have new tips mounted on them. If you have
saved the old bases, why don't you check with them to see if they still
do the service. There used to be quite a difference between retipping
and the price of a new cathode.

Cheers,
Henk
(no particular interest in EBS other than as a satisfied customer of
cathodes some time ago)


At 9/17/2010 6:28 PM, jkrupp-at-deltacollege.edu wrote:
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe --http://www.microscopy.com/MicroscopyListserver
} On-Line Helphttp://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi
}
} Some of you old timers may remember the venerable ETEC SEM. We have
} one we use for training students hands on for filament exchange etc.
} The scope no longer works, but we keep the column so students can
} practice.
}
} Part of their training involves cleaning the gun parts and
} reassembling the filament, grid cap, etc. We had a stash of old
} filaments they used to practice centering, but today two of our supply
} were broken. This exhausts our supply so we are asking if anyone has
} some old ETEC filaments in a drawer that you could send our way, or
} ideas about substitutes we could use for this exercise.
}
} Thanks
}
} Jon
}
} Jonathan Krupp
} Delta College
} 5151Pacific Ave.
} Stockton, CA 95207
} 209-954-5284
} jkrupp-at-deltacollege.edu
}
}
}
}
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}

--
Hendrik O. Colijn www.ceof.ohio-state.edu
OSU Campus Electron Optics Facility colijn.1-at-osu.edu
040 Fontana Labs (614) 292-0674 (V)
116 W. 19th Ave. (614) 292-7523 (F)
Columbus, OH 43210

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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From: david.knecht-at-uconn.edu
Date: Sat, 18 Sep 2010 07:36:35 -0500
Subject: [Microscopy] Laser catapulting

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

We read a paper in journal club today that used a laser catapulting technique. They claim to be able to eject a chunk of tissue off of a glass surface, out of the solution and into the cap of a microtube using a pulse of UV laser and at the same time cause no heating. " the sample is driven with high speed along the wave front of the powerful photonic stream and can be "beamed" several millimeters away, even against gravity" Apparently it works, but sounded like science fiction to me. Is this something like what happens in a laser tweezer, but instead of a trapping cone, you have a linear force produced? Could that really be strong enough to break surface tension?
http://palm.dasat.ch/dasat/index.php?cid=100147&sid=dasat

Dr. David Knecht
Department of Molecular and Cell Biology
Co-head Flow Cytometry and Confocal Microscopy Facility
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)




==============================Original Headers==============================
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From: kenconverse-at-qualityimages.biz
Date: Sat, 18 Sep 2010 09:06:42 -0500
Subject: [Microscopy] ETEC filaments?

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

So John,
You're saying I'm a really old fart, huh? I was an ETEC customer in '75 and
went to work for them as a field engineer in '77. It was a terrific
company, but I have to sometimes remind myself not to get too mad at
Perkin-Elmer. If it weren't for P-E trashing ETEC, I never could have made
Quality Images work. Sometimes that's the way it goes.

As for the Master Anode Power Supply, you were a member of the "MAPS of the
Month Club". We in the field kept trying to tell Engineering that some
small changes needed to be made. Of course, we didn't know what we were
talking about, right? After I started Quality Images, I made those small
changes and the failures ended. Saved me a bundle!

Even 5 years ago ETECs made up half my business. Not bad considering the
last one was built in '82. Sadly, that's no longer true although I just
installed an Autoscan in our local middle school for the MS and HS kids to
play with. My kids' HS in PA has had one for 10 years, now, and it's used
constantly.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: John Mardinly [mailto:John.Mardinly-at-wdc.com]
Sent: Friday, September 17, 2010 9:58 PM
To: kenconverse-at-qualityimages.biz
Cc: Microscopy-at-microscopy.com

Jon,
I don't really like to think of myself as an "oldtimer", but maybe that's
just vanity.

First, why isn't your ETEC Autoscan still running? It should be. It's what
I consider to be the most student-proof SEM available.

Second, I believe Gene Taylor still has ETEC filaments available.
http://www.semicro.org/search.aspx?find=etec+filaments

I'm glad he does because I have ETEC customers still running their systems
on a daily basis. Why isn't yours running?

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: jkrupp-at-deltacollege.edu [mailto:jkrupp-at-deltacollege.edu]
Sent: Friday, September 17, 2010 6:31 PM
To: kenconverse-at-qualityimages.biz

Hi

Some of you old timers may remember the venerable ETEC SEM. We have
one we use for training students hands on for filament exchange etc.
The scope no longer works, but we keep the column so students can
practice.

Part of their training involves cleaning the gun parts and
reassembling the filament, grid cap, etc. We had a stash of old
filaments they used to practice centering, but today two of our supply
were broken. This exhausts our supply so we are asking if anyone has
some old ETEC filaments in a drawer that you could send our way, or
ideas about substitutes we could use for this exercise.

Thanks

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu




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From: sarj0007-at-unf.edu
Date: Sun, 19 Sep 2010 14:55:02 -0500
Subject: [Microscopy] Thin section thickness guides and sorvall MT2

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

Henk,
That would probably work fine if Jon isn't going to get the system running,
but my experience is that most of the suppliers don't have the precision in
mounting required for ETEC. There is no means to change the alignment of
the filament to the grid (wehnelt) cap, as there is on virtually every other
SEM. This greatly simplifies filament changes, but requires great precision
in mounting the tungsten wire to the posts.

X and Y are +/- .002" and the height is +/- .0007". ETEC made the
measurements AFTER burning in the filaments to stress-relieve them. No one
does the stress relief now, and I've found that Gene Taylor has the most
consistent filaments since ETEC quit making them.

In fact, ETEC was so consistent that if you installed a filament and it was
off-center, the whole rest of the box was also off-center. It didn't happen
very often, but once in a while...

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: colijn.1-at-osu.edu [mailto:colijn.1-at-osu.edu]
Sent: Friday, September 17, 2010 10:45 PM
To: kenconverse-at-qualityimages.biz

Hi Jonathan,

In the "old days" we used to send our burned out filament bases to
Energy Beam Sciences to have new tips mounted on them. If you have
saved the old bases, why don't you check with them to see if they still
do the service. There used to be quite a difference between retipping
and the price of a new cathode.

Cheers,
Henk
(no particular interest in EBS other than as a satisfied customer of
cathodes some time ago)


At 9/17/2010 6:28 PM, jkrupp-at-deltacollege.edu wrote:
}
}
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} Hi
}
} Some of you old timers may remember the venerable ETEC SEM. We have
} one we use for training students hands on for filament exchange etc.
} The scope no longer works, but we keep the column so students can
} practice.
}
} Part of their training involves cleaning the gun parts and
} reassembling the filament, grid cap, etc. We had a stash of old
} filaments they used to practice centering, but today two of our supply
} were broken. This exhausts our supply so we are asking if anyone has
} some old ETEC filaments in a drawer that you could send our way, or
} ideas about substitutes we could use for this exercise.
}
} Thanks
}
} Jon
}
} Jonathan Krupp
} Delta College
} 5151Pacific Ave.
} Stockton, CA 95207
} 209-954-5284
} jkrupp-at-deltacollege.edu
}
}
}
}
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Message-Id: {B9606438-C195-414D-9B84-F51F1E0A400E-at-deltacollege.edu}
} 9, 36 -- From: Jon Krupp {jkrupp-at-deltacollege.edu}
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} 9, 36 -- Date: Fri, 17 Sep 2010 15:27:18 -0700
} 9, 36 -- X-Mailer: Apple Mail (2.936)
} ==============================End of -
Headers==============================
}

--
Hendrik O. Colijn www.ceof.ohio-state.edu
OSU Campus Electron Optics Facility colijn.1-at-osu.edu
040 Fontana Labs (614) 292-0674 (V)
116 W. 19th Ave. (614) 292-7523 (F)
Columbus, OH 43210

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

==============================Original Headers==============================
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From ahx-at-casablanca-siegen.de Sat Sep 18 16:32:07 2010
Return-Path: {ahx-at-casablanca-siegen.de}
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Received: from [190.76.127.61] by mxlb.ispgateway.de; Sat, 18 Sep 2010 16:27:43 -0500

Hello again. I wanted to thank everyone for the numerous replies on both issues, especially Mrs. Connelly’s wonderfully detailed notes she sent off listserv. Aside from some tweaking the o-ring fixed the problem and now I just need to learn how to section properly… And I have to agree that these machines are really wonderfully put together. People just don’t build a lot of things like they used to anymore.

I believe I have found a few sources people mentioned to try on Monday when offices open up for the color chart. The problem is the color chart my professor used stayed in their respective laboratories when she left them (many years ago), and we are somewhat attempting to put things together from scratch here.

I agree it would be nice to see something a little more readily available from an online source. Also, some people have some very amusing out of office replies. It’s worth the flood of them that come. Enjoy the last bit of the weekend.

-Jason Saredy


________________________________________
X-from: bozzola-at-siu.edu [bozzola-at-siu.edu]
Sent: Friday, September 17, 2010 1:46 PM
To: Saredy, Jason

Wouldn't it be great to see something like this in Microscopy Today?
Hint, hint, hint .....

--

John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL 62901
Phone: 618-453-3730


On Fri, Sep 17, 2010 at 11:16 AM, Factor, Jan {Jan.Factor-at-purchase.edu} wrote:
} If John Bozzola is correct, and the color chart is not under copyright, then
} perhaps someone who has a copy that is still in good condition would be
} willing to scan it and make it available to the list. Just a thought...
} --Best, Jan Factor
}
} Jan Robert Factor, Ph.D.
} Professor of Biology
} Purchase College, State University of New York
} Purchase, NY 10577
} Office: 2016NS, 914-251-6659
} Office Hours (Fall 2010): Mon., 10:30-12:00, Wed., 4:30-6:00
} jan.factor-at-purchase.edu
}
} Please consider the environment before printing this e-mail
} ________________________________________
} From: bozzola-at-siu.edu [bozzola-at-siu.edu]
} Sent: Friday, September 17, 2010 11:52 AM
} To: Factor, Jan
} Subject: [Microscopy] Re: Thin section thickness guides and sorvall MT2
}
} ----------------------------------------------------------------------------
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} Regarding the laminated, section-thickness, color scale distributed by
} Sorvall, often given away with their ultramicrotomes. This was based
} on the work of Lee Peachey.
}
} I don't believe the original is copyrighted (but check beforehand), so
} why not borrow the original from your professor and scan it using a
} high quality, color scanner?
}
} John Bozzola
} IMAGE Center
} Southern Illinois University
} Carbondale, IL
}
} ==============================Original Headers==============================
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} MT2
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From: max_histo_00-at-yahoo.it
Date: Mon, 20 Sep 2010 06:56:55 -0500
Subject: [Microscopy] viaWWW: Laboratory procedure - histological preparations

Contents Retrieved from Microscopy Listserver Archives
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Email: max_histo_00-at-yahoo.it
Name: Massimo Tosi

Organization: Private

Title-Subject: [Filtered] Laboratory procedure

Message: Hi all,

I found into a shelf of the laboratory a small flask containing mouse samples
fixed in Bouin's fluid and preserved in ethanol at 70ƒ, forgotten there for a
few years.
I wonder if it would be possible to continue the process up to paraffin
embedding for histological preparations.
Some time ago a professor of biology at the University of Florence told me that
they could remain in alcohol for years, but on literature I found that the time
can not be so long without altering tissues.
Has anyone had a similar experience?
It's better to throw everything away or not?
Thank you in advance.

Kind Regards,

Massimo Tosi

Login Host: 151.57.0.253
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From: beth-at-plantbio.uga.edu
Date: Mon, 20 Sep 2010 10:36:39 -0500
Subject: [Microscopy] Re: Laser catapulting

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

We have the Zeiss PALM (Position and Ablation with Laser Microbeams)
system in our lab. A pulsed ultra-violet (UV-A) laser beam is
interfaced into the microscope and focused through an objective lens
to a beam spot size of less than 1 µm in diameter for the sample
cutting action. After microdissection of tissue on special membrane
slides isolated cells are ejected out of the object plane and
catapulted directly into the cap of a common microfuge tube. This is
performed in an entirely non-contact manner with the help of a single
defocused laser pulse.

It is a bit different when the tissue is only on glass slides. After
outlining your area of interest you can only use an auto-LPC mode - in
this mode the software fills in the area of the tissue you have
circled with dots and the laser pulses each dot catapulting bits of
the tissue into the eppendorf cap. It's like watching Bonnie & Clyde
or The Terminator machine gun the sample. Kinda cool. If you have the
laser set too high it can etch the glass slide.

Beth

On Sep 18, 2010, at 8:37 AM, david.knecht-at-uconn.edu wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
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} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} We read a paper in journal club today that used a laser catapulting
} technique. They claim to be able to eject a chunk of tissue off of a
} glass surface, out of the solution and into the cap of a microtube
} using a pulse of UV laser and at the same time cause no heating. "
} the sample is driven with high speed along the wave front of the
} powerful photonic stream and can be "beamed" several millimeters
} away, even against gravity" Apparently it works, but sounded like
} science fiction to me. Is this something like what happens in a
} laser tweezer, but instead of a trapping cone, you have a linear
} force produced? Could that really be strong enough to break surface
} tension?
} http://palm.dasat.ch/dasat/index.php?cid=100147&sid=dasat
}
} Dr. David Knecht
} Department of Molecular and Cell Biology
} Co-head Flow Cytometry and Confocal Microscopy Facility
} U-3125
} 91 N. Eagleville Rd.
} University of Connecticut
} Storrs, CT 06269
} 860-486-2200
} 860-486-4331 (fax)
}
}
}
}
} ==============================Original
} Headers==============================
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From: maloneyb-at-fiu.edu
Date: Mon, 20 Sep 2010 10:43:50 -0500
Subject: [Microscopy] TEM prep

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Dear Group - has anyone ever used cells grown on hydrogel and imaged in
TEM? If so, would you be so kind as to forward to me details.
Thanks so much.
Barbara

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From: kraftpiano-at-gmail.com
Date: Mon, 20 Sep 2010 11:04:13 -0500
Subject: [Microscopy] Laser catapulting

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Not to make light of an extremely interesting topic (I am a physicist by nature...) but a colleague of mine and I were just commenting on how this would make by far the geekiest game of quarters we could possibly conceive.

"He got the mitochondria right into the pollen spore! Everybody drink!"

--Justin A. Kraft

On Sep 20, 2010, at 11:43 AM, beth-at-plantbio.uga.edu wrote:

}
}
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} We have the Zeiss PALM (Position and Ablation with Laser Microbeams)
} system in our lab. A pulsed ultra-violet (UV-A) laser beam is
} interfaced into the microscope and focused through an objective lens
} to a beam spot size of less than 1 µm in diameter for the sample
} cutting action. After microdissection of tissue on special membrane
} slides isolated cells are ejected out of the object plane and
} catapulted directly into the cap of a common microfuge tube. This is
} performed in an entirely non-contact manner with the help of a single
} defocused laser pulse.
}
} It is a bit different when the tissue is only on glass slides. After
} outlining your area of interest you can only use an auto-LPC mode - in
} this mode the software fills in the area of the tissue you have
} circled with dots and the laser pulses each dot catapulting bits of
} the tissue into the eppendorf cap. It's like watching Bonnie & Clyde
} or The Terminator machine gun the sample. Kinda cool. If you have the
} laser set too high it can etch the glass slide.
}
} Beth
}
} On Sep 18, 2010, at 8:37 AM, david.knecht-at-uconn.edu wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------------
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} }
} } We read a paper in journal club today that used a laser catapulting
} } technique. They claim to be able to eject a chunk of tissue off of a
} } glass surface, out of the solution and into the cap of a microtube
} } using a pulse of UV laser and at the same time cause no heating. "
} } the sample is driven with high speed along the wave front of the
} } powerful photonic stream and can be "beamed" several millimeters
} } away, even against gravity" Apparently it works, but sounded like
} } science fiction to me. Is this something like what happens in a
} } laser tweezer, but instead of a trapping cone, you have a linear
} } force produced? Could that really be strong enough to break surface
} } tension?
} } http://palm.dasat.ch/dasat/index.php?cid=100147&sid=dasat
} }
} } Dr. David Knecht
} } Department of Molecular and Cell Biology
} } Co-head Flow Cytometry and Confocal Microscopy Facility
} } U-3125
} } 91 N. Eagleville Rd.
} } University of Connecticut
} } Storrs, CT 06269
} } 860-486-2200
} } 860-486-4331 (fax)
} }
} }
} }
} }
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From: dainis-at-dauksta.com
Date: Mon, 20 Sep 2010 11:36:30 -0500
Subject: [Microscopy] viaWWW: Bubbles in Newport mx 530 micromanipulator

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Email: dainis-at-dauksta.com
Name: Dainis Dauksta

Title-Subject: [Filtered] Bubbles in Newport mx 530 micromanipulator

Message: Hallo, Does anyone know if there any companies who repair
and refill Newport mx 530 hydraulic micromanipulators please?
Thanks
Dainis

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From: goodlg-at-matthey.com
Date: Mon, 20 Sep 2010 12:07:39 -0500
Subject: [Microscopy] Vacancy: SEM/TEM (UK)

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VACANCY: JOHNSON MATTHEY
Technology Centre - Sonning Common (approx. 35miles West of Central
London)

ELECTRON MICROSCOPIST

Johnson Matthey is a speciality chemicals company focused on its core
skills in
catalysis, precious metals, fine chemicals and process technology. The

Technology Centre, based at Sonning Common, undertakes research work
for the group.

A vacancy has arisen for an Electron Microscopist to join our team
working
with both Scanning and Transmission microscopes. The successful
candidate
is expected to have experience in several of the following areas of
expertise:

SEM and TEM characterisation methods: (HRTEM; STEM; HAADF Imaging;
EDX; EELS, Electron diffraction analysis, Cs corrected imaging)

Electron Microscope technical knowledge: (FEG sources, vacuum system;
electronics, fault finding and diagnosis)

SEM and TEM sample preparation: (Coating; Ultramicrotomy; Cryo-sample
preparation)

This job is based on the analysis of a variety of samples from our
research
groups and various divisions, thus prior exposure to multi-disciplinary
subjects,
catalysis and materials as well as computer programming and scripting
ability
would be an advantage.

The successful candidate will be educated to HNC/Degree level as a
minimum.
Applications must be made in writing with full CV and current salary
details to:
Georgie Floyd, Personnel Officer, Johnson Matthey Technology Centre,
Blounts Court, Sonning Common, Reading RG4 9NH
or e-mail: hrjmtc-at-matthey.com

CLOSING DATE FOR APPLICATIONS: 1st October 2010

Johnson Matthey Plc is an equal opportunities employer and positively
encourages applications form suitably qualified and eligible
candidates
regardless of sex, race, disability, age, sexual orientation, religion
or belief.


If the reader of this email is not the intended recipient(s), please be advised that any dissemination, distribution or copying of this information is strictly prohibited. Johnson Matthey PLC has its main place of business at 40-42 Hatton Garden, London (020 7269 8400).

Johnson Matthey Public Limited Company
Registered Office: 40-42 Hatton Garden, London EC1N 8EE
Registered in England No 33774

Whilst Johnson Matthey aims to keep its network free from viruses you should note that we are unable to scan certain emails, particularly if any part is encrypted or password-protected, and accordingly you are strongly advised to check this email and any attachments for viruses. The company shall NOT ACCEPT any liability with regard to computer viruses transferred by way of email.

Please note that your communication may be monitored in accordance with Johnson Matthey internal policy documentation.

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From: dsherman-at-purdue.edu
Date: Mon, 20 Sep 2010 12:11:56 -0500
Subject: [Microscopy] Objective lens in SEM?

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Hi all,

Many of you will agree that what is termed as 'objective lens" in an SEM is
more aptly named "final lens". My interpretation of an objective lens is
one that magnifies an image such as in a light microscope whereas the final
lens in an SEM is actually a reducing lens as are all the other lenses in
the SEM column.

A number of years ago (1994) there was an article in "Microscopy Today" that
supported this definition of this last lens as a FINAL lens or it actually
could be considered as a 3rd condenser lens. I agreed with that
interpretation. However I disagree with what came next and would like your
input on the interpretation.

The author went on to say that ³the Abbe equation (0.612 (constant) x
wavelength/refractive index of vacuum x the sin of the half angle of
illumination) , as it refers to the half-angle of illumination, applies only
to objective lenses and thus did not apply to any lenses in an SEM column.
Therefore it cannot be used to ³calculate² resolution. The only thing that
determines resolution is spot size, not wavelength or accelerating voltage.²

My take is that the shorter the electron wavelength, the more energetic the
electrons and the faster they travel. This would act to reduce the effects
of spherical and chromatic aberration that would act to decrease potential
resolution. In addition, strength of the condenser lenses determines the
position of crossover below each lens, and thus the size of the resulting
Airy disk (spot size) and coherence of resulting beam as well as probe
strength. In the case of the final lens, the cross-over must impinge on the
surface of the sample for the sample to be in focus. Thus working distance
will come into play with the closer the sample is to the bottom of the lens,
the greater is the 1/2 angle of illumination and thus the better the
resolution (Abbe equation).

Of course this is simplistic in that the ultimate resolution depends on many
other factors, such as type of sample, ultimate design and capability of the
instrument, effects of diffraction along with chromatic and spherical
aberration, parameters used for the imaging, and environmental factors in
the vicinity of the microscope. But can the Abbe equation be used to predict
the limit of resolution in the ideal case...primarily as a teaching tool
even if not totally accurate numerically?

Comments please.....

Debby


---
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy

--
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy/



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From: protrain-at-emcourses.com
Date: Mon, 20 Sep 2010 14:08:13 -0500
Subject: [Microscopy] Objective lens in SEM?

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

Hi All

Debby has raised interesting points worthy of discussion.

Firstly we should ask "what is an objective lens?" An objective lens
CREATES an image. Which part of a SEM creates an image, well it is the
electronics that create! So the final lens in the SEM should not be called
an objective lens it is simply a third condenser lens.

Now for part two. Throughout my career I have been in the position of
having to find more resolution from an SEM and the battle has always been in
two parts, against the poor quality of a tungsten hairpin source and against
chromatic aberration.

If the source has a high brightness the condenser system has more beam to
work with which means making the probe smaller is not such a problem; FEG
SEM are good examples of this.

The tungsten hairpin source was a problem because its brightness was often
insufficient, the source was too large and too dim, so we moved filaments
forward, sharpened filaments and even used pointed filaments, then LaB6 came
along. Straight away more performance because Lab6 had a smaller source
size therefore higher brightness and helping the chromatic effects it used
less heat. But in both these cases, as we mentioned brightness, we do have
an angular effect = amps/sqcm/steradian

Down the other end of the column we moved the specimen as close to the lens
as possible to increase the lens current thus minimise chromatic aberration;
sure there are angles but I believe unimportant angles. Then I became
involved with the ISI/Akashi project the development of the first "in lens
detection" SEM; even higher lens currents and even less chromatic
aberration. Add to this the pressure applied to manufacturers to re design
their SEM lenses with aberration coefficients suitable for lower
accelerating voltages and suddenly we had maximum performance at 15kV rather
than 30!

Look at what the SEM manufacturers are doing. Many are bringing the beam
down the column at a medium accelerating voltage then reducing its energy,
lowering the accelerating voltage at the last moment as the beam leaves the
final lens. The big plus is the aberration reduction, the lenses in the
column run at higher currents reducing chromatic aberration. One
manufacturer has even removed a condenser lens from the system changing the
probe current through apertures rather than lenses. Have they hit on a trick
to reduce chromatic aberration by simply removing a lens; less aberration!

So my simple interpretation of the practical effects that I have seen is as
follows - the performance of a SEM depends upon the brightness of the source
along with the chromatic aberration; the sum of all the stabilities. Try
running a SEM flat out within 10 minutes of switching the high voltage on
and you will see the instabilities for yourself.

Have fun

Steve


Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com

-----Original Message-----
X-from: dsherman-at-purdue.edu [mailto:dsherman-at-purdue.edu]
Sent: 20 September 2010 18:13
To: protrain-at-emcourses.com

Hi all,

Many of you will agree that what is termed as 'objective lens" in an SEM is
more aptly named "final lens". My interpretation of an objective lens is
one that magnifies an image such as in a light microscope whereas the final
lens in an SEM is actually a reducing lens as are all the other lenses in
the SEM column.

A number of years ago (1994) there was an article in "Microscopy Today" that
supported this definition of this last lens as a FINAL lens or it actually
could be considered as a 3rd condenser lens. I agreed with that
interpretation. However I disagree with what came next and would like your
input on the interpretation.

The author went on to say that ³the Abbe equation (0.612 (constant) x
wavelength/refractive index of vacuum x the sin of the half angle of
illumination) , as it refers to the half-angle of illumination, applies only
to objective lenses and thus did not apply to any lenses in an SEM column.
Therefore it cannot be used to ³calculate² resolution. The only thing that
determines resolution is spot size, not wavelength or accelerating voltage.²

My take is that the shorter the electron wavelength, the more energetic the
electrons and the faster they travel. This would act to reduce the effects
of spherical and chromatic aberration that would act to decrease potential
resolution. In addition, strength of the condenser lenses determines the
position of crossover below each lens, and thus the size of the resulting
Airy disk (spot size) and coherence of resulting beam as well as probe
strength. In the case of the final lens, the cross-over must impinge on the
surface of the sample for the sample to be in focus. Thus working distance
will come into play with the closer the sample is to the bottom of the lens,
the greater is the 1/2 angle of illumination and thus the better the
resolution (Abbe equation).

Of course this is simplistic in that the ultimate resolution depends on many
other factors, such as type of sample, ultimate design and capability of the
instrument, effects of diffraction along with chromatic and spherical
aberration, parameters used for the imaging, and environmental factors in
the vicinity of the microscope. But can the Abbe equation be used to predict
the limit of resolution in the ideal case...primarily as a teaching tool
even if not totally accurate numerically?

Comments please.....

Debby


---
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy

--
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy/



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From: oshel1pe-at-cmich.edu
Date: Mon, 20 Sep 2010 14:10:31 -0500
Subject: [Microscopy] Re: Objective lens in SEM?

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

Debby,

My take (what we teach):
1) The final lens is just that, aka last condenser lens. Not an objective lens.
2) The Abbe equation doesn't apply because the
SEM is not an image forming instrument. It is a
signal generator/detector.
The interaction of the beam with the
specimen generates a signal: secondary and
primary (backscattered) electrons, Auger
electrons, x-rays, induced current, etc.. These
signals are detected as a function of the beam
position, but they are not detected in any form
that forms an image. Rather, the resultant image
is simply a construction made by mapping signal
intensity again as a function of beam position.
Abbe is irrelevant to this process. (Like how an
image is formed on a TV tube, or computer
monitor.)

Your comments about aberrations (chromatic and
spherical) and lenses and the Abbe equation are I
think correct. The electron lens acts as an
image-forming lens, and the SEM column above the
sample is the same thing as a TEM column above
the sample. So optical equations and ray-tracing
apply. It's how the beam is used that differs.
Scanning a spot and collecting generated signals
versus illuminating the sample and forming an
image in an image plane.

So, shorter wavelength (higher energy) electrons
can form higher resolution images in a SEM
because it's easier to reduce aberrations, and
aberrations cause a non-round, smeared-out spot
where the beam impacts the specimen. But, lower
energy electrons can actually form a higher
resolution energy because the signal is generated
from a smaller volume.

Practical example: the Hitachi 5500 (if I
remember the model number right) in-lens FE-SEM
has a resolution of at least 0.5 nanometers at
30kV in transmission mode, and about 1.4
nanometers at 1kV in scanning mode. It's
optimised for maximum resolution in transmission
(like a STEM). The 4800 below-lens FE-SEM has 1
nm resolution at 1kV in scanning mode (if I
remember the numbers right - it is better is the
point), because it is optimised for low-kV spot
scanning.
The point being, higher resolution is achieved
with 1kV than is achieved with 30kV - lower
energy rather than higher energy - because a
smaller scanning spot at a lower energy is
obtained.

Smaller scanning spot + lower kV = smaller
beam-specimen interaction volume, therefore the
signal (secondary electrons, etc.) is generated
from a smaller volume, therefore higher
resolution.
Abbe doesn't matter here. So no, the Abbe
relation can't be used to calculate resolution in
the SEM.

The question then is "how is the smaller spot
size achieved?" Here, electron optics and ray
tracing and Abbe do matter. Sort of.
Have you seen this web page:
http://chemgroups.northwestern.edu/odom/nano-characterization/SEM%20Lab1-fl.htm
Scroll down to "Flash tutorial on the effect of
condenser lens in controlling resolution".

The basic point is Abbe is relevant to optical,
aka image-forming, systems like light microscopes
and TEMs. It is only relevant in a more limited
way in non-optical, aka non-image forming
systems, like SEMs and AFMs.
It's easier to think of a microscope taxonomy:
SEM is in the same clade as AFM, not TEM. TEM is
image forming like light microscopy, SEM is
scanned-probe like AFM. It just uses a beam of
electrons for its probe (instead of a mechanical
cantilever), and Abbe, etc., is of relevance in
determining the formation of that probe - but not
the image.

Phil

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From: bozzola-at-siu.edu
Date: Mon, 20 Sep 2010 14:37:20 -0500
Subject: [Microscopy] SEM: filament image

Contents Retrieved from Microscopy Listserver Archives
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In the TEM, we saturate a conventional, tungsten filament based on
appearance of the electron cloud surrounding the filament tip. In the
SEM, filament saturation is achieved by observing the brightness
displayed as a line profile.

Our 28 yr old Hitachi S-570 SEM has the capability to generate an
image of the filament tip, just like a TEM. I've never heard a good
explanation of how this is done. Does anyone know?

Thanks.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


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From: sneoh-at-eaglabs.com
Date: Tue, 21 Sep 2010 06:29:49 -0500
Subject: [Microscopy] viaWWW: Looking for leads on SAM Engineer

Contents Retrieved from Microscopy Listserver Archives
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I don't see why the term objective is such a problem: it is the lens that
sits around the *object* under study, whether it is a TEM, STEM or SEM. The
term 'final' suggests the last lens in a series which could be anything
from the projector lens in a TEM, the gun lens in a VG STEM or the
objective lens in a SEM (here my definition of final is the lens sitting
closest to the ground). Objective seems quite appropriate given the variety
of definitions one could use.

The Abbe equation you refer to refers to the smallest lateral space that
monochromatic radiation can be brought in to given by the limited solid
angle of the final/objective aperture (the Diffraction limit). It assumes
the source of radiation is infinitesimal and this is important- it gives
you the smallest size of the probe even if you found the most amazing
electron source in the universe (not including entangled sources, see
below).

Given this limitation (by the laws of physics), once you factor in the
aberrations and the fact that the source is extended, you get a
finite-current-carrying probe size that is much bigger- one that is
determined by balancing the aberrations with the diffraction limit. The Log
(current, I) versus Log (probe size) plots in figure 2.26 of Goldsteins
book "SEM and X-ray microanalysis" (3rd Ed, 2003) showing the probe current
vs probe size is precisely this. The asymptote (I -} 0) gives the probe
size given by balancing the aberrations and the diffraction limit for an
infinitely small source and is given by ~(Cs wavelength^3)^(1/4), for a
non-aberration corrected SEM.

The focused probe is the one the each EM manufacturer has control over and
actually measuring it is pretty difficult. Given the atomic nature of
normal (baryonic) matter, you can't get an infinitely sharp edge to scan
the beam over to get a probe shape profile. Once the electron enters the
sample, all hell breaks loose and the resolution you get varies
tremendously. But that's another story.

Note: for quantum entangled sources you get better resolution. For an
N-fold entangled source, the diffraction limit becomes ~wavelength/(N
semi-angle). Optical lithographers have started playing with these to
reduce feature size in semiconductors, but I don't see any hope of getting
a 2-fold entangled electron source any time soon, let alone N-fold (N} 2)...

Jonathan
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} Hi all,
}
} Many of you will agree that what is termed as 'objective lens" in an SEM is
} more aptly named "final lens". My interpretation of an objective lens is
} one that magnifies an image such as in a light microscope whereas the final
} lens in an SEM is actually a reducing lens as are all the other lenses in
} the SEM column.
}
} A number of years ago (1994) there was an article in "Microscopy Today"
} that supported this definition of this last lens as a FINAL lens or it
} actually could be considered as a 3rd condenser lens. I agreed with that
} interpretation. However I disagree with what came next and would like
} your input on the interpretation.
}
} The author went on to say that �the Abbe equation (0.612 (constant) x
} wavelength/refractive index of vacuum x the sin of the half angle of
} illumination) , as it refers to the half-angle of illumination, applies
} only to objective lenses and thus did not apply to any lenses in an SEM
} column. Therefore it cannot be used to �calculate� resolution. The only
} thing that determines resolution is spot size, not wavelength or
} accelerating voltage.�
}
} My take is that the shorter the electron wavelength, the more energetic
} the electrons and the faster they travel. This would act to reduce the
} effects of spherical and chromatic aberration that would act to decrease
} potential resolution. In addition, strength of the condenser lenses
} determines the position of crossover below each lens, and thus the size
} of the resulting Airy disk (spot size) and coherence of resulting beam as
} well as probe strength. In the case of the final lens, the cross-over
} must impinge on the surface of the sample for the sample to be in focus.
} Thus working distance will come into play with the closer the sample is
} to the bottom of the lens, the greater is the 1/2 angle of illumination
} and thus the better the resolution (Abbe equation).
}
} Of course this is simplistic in that the ultimate resolution depends on
} many other factors, such as type of sample, ultimate design and
} capability of the instrument, effects of diffraction along with chromatic
} and spherical aberration, parameters used for the imaging, and
} environmental factors in the vicinity of the microscope. But can the Abbe
} equation be used to predict the limit of resolution in the ideal
} case...primarily as a teaching tool even if not totally accurate
} numerically?
}
} Comments please.....
}
} Debby
}
}
} ---
} Debby Sherman, Director Phone: 765-494-6666
} Life Science Microscopy Facility FAX: 765-494-5896
} Purdue University E-mail: dsherman-at-purdue.edu
} S-052 Whistler Building
} 170 S. University Street
} West Lafayette, IN 47907
} http://www.ag.purdue.edu/facilities/microscopy
}
}


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From: bozzola-at-siu.edu
Date: Tue, 21 Sep 2010 08:04:45 -0500
Subject: [Microscopy] Objective lens in SEM?

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

Interesting discussion of lens nomenclature in an SEM.

I understand why some would use the term "objective lens," since it
has been defined (using terms developed for light microscopy) as the
lens closest to the object, in contrast to an eyepiece lens. These
lenses have been defined by proximity to an object.

On the other hand, "condenser lens" defines the function of the lens.
So, it all depends upon what you want to convey (location of lens or
functionality). Personally, I prefer "final condenser lens," which
conveys both. Other lenses would be referred to as "C1" or "C2", etc.

I doubt that my blood pressure would rise significantly if someone
were to use the other terms.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901


On Tue, Sep 21, 2010 at 5:03 AM, {jsb43-at-cam.ac.uk} wrote:
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} I don't see why the term objective is such a problem: it is the lens that
} sits around the *object* under study, whether it is a TEM, STEM or SEM. The
} term 'final' suggests the last lens in a series which could be anything
} from the projector lens in a TEM, the gun lens in a VG STEM or the
} objective lens in a SEM (here my definition of final is the lens sitting
} closest to the ground). Objective seems quite appropriate given the variety
} of definitions one could use.
}
} The Abbe equation you refer to refers to the smallest lateral space that
} monochromatic radiation can be brought in to given by the limited solid
} angle of the final/objective aperture (the Diffraction limit). It assumes
} the source of radiation is infinitesimal and this is important- it gives
} you the smallest size of the probe even if you found the most amazing
} electron source in the universe (not including entangled sources, see
} below).
}
} Given this limitation (by the laws of physics), once you factor in the
} aberrations and the fact that the source is extended, you get a
} finite-current-carrying probe size that is much bigger- one that is
} determined by balancing the aberrations with the diffraction limit. The Log
} (current, I) versus Log (probe size) plots in figure 2.26 of Goldsteins
} book "SEM and X-ray microanalysis" (3rd Ed, 2003) showing the probe current
} vs probe size is precisely this. The asymptote (I -} 0) gives the probe
} size given by balancing the aberrations and the diffraction limit for an
} infinitely small source and is given by ~(Cs wavelength^3)^(1/4), for a
} non-aberration corrected SEM.
}
} The focused probe is the one the each EM manufacturer has control over and
} actually measuring it is pretty difficult. Given the atomic nature of
} normal (baryonic) matter, you can't get an infinitely sharp edge to scan
} the beam over to get a probe shape profile. Once the electron enters the
} sample, all hell breaks loose and the resolution you get varies
} tremendously. But that's another story.
}
} Note: for quantum entangled sources you get better resolution. For an
} N-fold entangled source, the diffraction limit becomes ~wavelength/(N
} semi-angle). Optical lithographers have started playing with these to
} reduce feature size in semiconductors, but I don't see any hope of getting
} a 2-fold entangled electron source any time soon, let alone N-fold (N} 2)...
}
} Jonathan
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe --
} } http://www.microscopy.com/MicroscopyListserver On-Line Help
} } http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } Hi all,
} }
} } Many of you will agree that what is termed as 'objective lens" in an SEM is
} } more aptly named "final lens".  My interpretation of an objective lens is
} } one that magnifies an image such as in a light microscope whereas the final
} } lens in an SEM is actually a reducing lens as are all the other lenses in
} } the SEM column.
} }
} } A number of years ago (1994) there was an article in "Microscopy Today"
} } that supported this definition of this last lens as a FINAL lens or it
} } actually could be considered as a 3rd condenser lens. I agreed with that
} } interpretation. However I disagree with what came next and would like
} } your input on the interpretation.
} }
} } The author went on to say that �the Abbe equation (0.612 (constant) x
} } wavelength/refractive index of vacuum x the sin of the half angle of
} } illumination) , as it refers to the half-angle of illumination, applies
} } only to objective lenses and thus did not apply to any lenses in an SEM
} } column. Therefore it cannot be used to �calculate� resolution. The only
} } thing that determines resolution is spot size, not wavelength or
} } accelerating voltage.�
} }
} } My take is that the shorter the electron wavelength, the more energetic
} } the electrons and the faster they travel. This would act to reduce the
} } effects of spherical and chromatic aberration that would act to decrease
} } potential resolution. In addition, strength of the condenser lenses
} } determines the position of crossover below each lens, and thus the size
} } of the resulting Airy disk (spot size) and coherence of resulting beam as
} } well as probe strength. In the case of the final lens, the cross-over
} } must impinge on the surface of the sample for the sample to be in focus.
} } Thus working distance will come into play with the closer the sample is
} } to the bottom of the lens, the greater is the 1/2 angle of illumination
} } and thus the better the resolution (Abbe equation).
} }
} } Of course this is simplistic in that the ultimate resolution depends on
} } many other factors, such as type of sample, ultimate design and
} } capability of the instrument, effects of diffraction along with chromatic
} } and spherical aberration, parameters used for the imaging, and
} } environmental factors in the vicinity of the microscope. But can the Abbe
} } equation be used to predict the limit of resolution in the ideal
} } case...primarily as a teaching tool even if not totally accurate
} } numerically?
} }
} } Comments please.....
} }
} } Debby
} }
} }
} } ---
} } Debby Sherman, Director               Phone: 765-494-6666
} } Life Science Microscopy Facility      FAX:  765-494-5896
} } Purdue University                     E-mail: dsherman-at-purdue.edu
} } S-052 Whistler Building
} } 170 S. University Street
} } West Lafayette, IN 47907
} } http://www.ag.purdue.edu/facilities/microscopy
} }
} }


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From: klivi-at-jhu.edu
Date: Tue, 21 Sep 2010 09:00:17 -0500
Subject: [Microscopy] Objective lens in SEM?

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

Since I teach both SEM and TEM methods, I have found that it is
important to describe the final lens of the SEM as a condenser lens
(or alternatively, the final probe-forming lens) so as not to confuse
those who use both techniques. (I stay away from the fact that the
objective lens in a CM series (S)TEM also can be used to form the
probe.) Another issue that tends to surprise students is that the
object of the SEM lenses is to DEMAGNIFY the image of the filament
(1st crossover). Magnification only happens when the scanned area is
mapped to the display. This is very different from the wide beam
method of conventional TEM.
Ken

|_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
Kenneth JT Livi, PhD
Director, The High-Resolution Analytical Electron Microbeam Facility
of the Integrated Imaging Center
Departments of Earth and Planetary Sciences and Biology
Olin Hall
3400 N Charles Street
Johns Hopkins University
Baltimore, Maryland 21218 USA
| | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| |
| :|









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From: frah0010-at-umn.edu
Date: Tue, 21 Sep 2010 10:10:41 -0500
Subject: [Microscopy] Objective lens in SEM?

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

I believe that the term "objective lens" simply derives from the fact that it is closest to the object (i.e., specimen) being observed. There isn't a function tied to its nomenclature, only its physical placement. In that sense, it should be perfectly acceptable to call the "final" lens in a SEM or electron microprobe an objective lens.

Best,
Ellery

-----------------
Ellery Frahm
Manager & Principal Analyst, Electron Microprobe Lab
Senior Research Fellow, Department of Geology & Geophysics
University of Minnesota - Twin Cities
http://probelab.geo.umn.edu










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From: maryflet-at-interchange.ubc.ca
Date: Tue, 21 Sep 2010 11:04:33 -0500
Subject: [Microscopy] Objective lens in SEM?

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

Dear List,
When I am describing the final lens on an SEM, I also call it the "focusing"
lens, since it is the one that focuses the beam on the sample, and
differentiates it from the condenser lens(es). Conveying the way that an SEM
magnifies an image is the trickiest part of SEM 101.

Regards,

Mary Fletcher
Electron Microscopist
Materials Eng. UBC
#309 - 6350 Stores Road
Vancouver, B.C. V6T 1Z4
Canada
Tel: 604-822-5648
Fax: 604-822-3619
email: maryflet-at-interchange.ubc.ca


-----Original Message-----
X-from: bozzola-at-siu.edu [mailto:bozzola-at-siu.edu]
Sent: September 21, 2010 6:15 AM
To: maryflet-at-interchange.ubc.ca

Interesting discussion of lens nomenclature in an SEM.

I understand why some would use the term "objective lens," since it
has been defined (using terms developed for light microscopy) as the
lens closest to the object, in contrast to an eyepiece lens. These
lenses have been defined by proximity to an object.

On the other hand, "condenser lens" defines the function of the lens.
So, it all depends upon what you want to convey (location of lens or
functionality). Personally, I prefer "final condenser lens," which
conveys both. Other lenses would be referred to as "C1" or "C2", etc.

I doubt that my blood pressure would rise significantly if someone
were to use the other terms.

--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901


On Tue, Sep 21, 2010 at 5:03 AM, {jsb43-at-cam.ac.uk} wrote:
}
}
}
}
----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of
America
} To  Subscribe/Unsubscribe --
http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
}
----------------------------------------------------------------------------
}
} I don't see why the term objective is such a problem: it is the lens that
} sits around the *object* under study, whether it is a TEM, STEM or SEM.
The
} term 'final' suggests the last lens in a series which could be anything
} from the projector lens in a TEM, the gun lens in a VG STEM or the
} objective lens in a SEM (here my definition of final is the lens sitting
} closest to the ground). Objective seems quite appropriate given the
variety
} of definitions one could use.
}
} The Abbe equation you refer to refers to the smallest lateral space that
} monochromatic radiation can be brought in to given by the limited solid
} angle of the final/objective aperture (the Diffraction limit). It assumes
} the source of radiation is infinitesimal and this is important- it gives
} you the smallest size of the probe even if you found the most amazing
} electron source in the universe (not including entangled sources, see
} below).
}
} Given this limitation (by the laws of physics), once you factor in the
} aberrations and the fact that the source is extended, you get a
} finite-current-carrying probe size that is much bigger- one that is
} determined by balancing the aberrations with the diffraction limit. The
Log
} (current, I) versus Log (probe size) plots in figure 2.26 of Goldsteins
} book "SEM and X-ray microanalysis" (3rd Ed, 2003) showing the probe
current
} vs probe size is precisely this. The asymptote (I -} 0) gives the probe
} size given by balancing the aberrations and the diffraction limit for an
} infinitely small source and is given by ~(Cs wavelength^3)^(1/4), for a
} non-aberration corrected SEM.
}
} The focused probe is the one the each EM manufacturer has control over and
} actually measuring it is pretty difficult. Given the atomic nature of
} normal (baryonic) matter, you can't get an infinitely sharp edge to scan
} the beam over to get a probe shape profile. Once the electron enters the
} sample, all hell breaks loose and the resolution you get varies
} tremendously. But that's another story.
}
} Note: for quantum entangled sources you get better resolution. For an
} N-fold entangled source, the diffraction limit becomes ~wavelength/(N
} semi-angle). Optical lithographers have started playing with these to
} reduce feature size in semiconductors, but I don't see any hope of getting
} a 2-fold entangled electron source any time soon, let alone N-fold
(N} 2)...
}
} Jonathan
} }
} }
} }
----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} } http://www.microscopy.com/MicroscopyListserver/FAQ.html
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} }
} } Hi all,
} }
} } Many of you will agree that what is termed as 'objective lens" in an SEM
is
} } more aptly named "final lens".  My interpretation of an objective lens is
} } one that magnifies an image such as in a light microscope whereas the
final
} } lens in an SEM is actually a reducing lens as are all the other lenses in
} } the SEM column.
} }
} } A number of years ago (1994) there was an article in "Microscopy Today"
} } that supported this definition of this last lens as a FINAL lens or it
} } actually could be considered as a 3rd condenser lens. I agreed with that
} } interpretation. However I disagree with what came next and would like
} } your input on the interpretation.
} }
} } The author went on to say that �the Abbe equation (0.612 (constant) x
} } wavelength/refractive index of vacuum x the sin of the half angle of
} } illumination) , as it refers to the half-angle of illumination, applies
} } only to objective lenses and thus did not apply to any lenses in an SEM
} } column. Therefore it cannot be used to �calculate� resolution. The
only
} } thing that determines resolution is spot size, not wavelength or
} } accelerating voltage.�
} }
} } My take is that the shorter the electron wavelength, the more energetic
} } the electrons and the faster they travel. This would act to reduce the
} } effects of spherical and chromatic aberration that would act to decrease
} } potential resolution. In addition, strength of the condenser lenses
} } determines the position of crossover below each lens, and thus the size
} } of the resulting Airy disk (spot size) and coherence of resulting beam as
} } well as probe strength. In the case of the final lens, the cross-over
} } must impinge on the surface of the sample for the sample to be in focus.
} } Thus working distance will come into play with the closer the sample is
} } to the bottom of the lens, the greater is the 1/2 angle of illumination
} } and thus the better the resolution (Abbe equation).
} }
} } Of course this is simplistic in that the ultimate resolution depends on
} } many other factors, such as type of sample, ultimate design and
} } capability of the instrument, effects of diffraction along with chromatic
} } and spherical aberration, parameters used for the imaging, and
} } environmental factors in the vicinity of the microscope. But can the Abbe
} } equation be used to predict the limit of resolution in the ideal
} } case...primarily as a teaching tool even if not totally accurate
} } numerically?
} }
} } Comments please.....
} }
} } Debby
} }
} }
} } ---
} } Debby Sherman, Director               Phone: 765-494-6666
} } Life Science Microscopy Facility      FAX:  765-494-5896
} } Purdue University                     E-mail:
dsherman-at-purdue.edu
} } S-052 Whistler Building
} } 170 S. University Street
} } West Lafayette, IN 47907
} } http://www.ag.purdue.edu/facilities/microscopy
} }
} }


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From: jehrman-at-mta.ca
Date: Tue, 21 Sep 2010 11:15:54 -0500
Subject: [Microscopy] Objective lens in SEM?

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

If you're lucky enough to have a microprobe or SEM with an optical
microscope, demonstrating magnification in an SEM is really slick. Put
the scan on a piece of polished benitoite or other fluorescing mineral,
and have the student look through the optical scope while playing with
the magnification and scan speed. It's mind-blowing the first time you
see it and drives home the 1-to-1 relationship between the scan on the
specimen to the display scan. You can also see the flyback retrace and
other things that have an effect on imaging and specimen contamination.
Adjusting focus is cool too, which also helps to show what the
"objective" lens does.

Jim

--

James M. Ehrman
Digital Microscopy Facility
Mount Allison University
63B York St.
Sackville, NB E4L 1G7
CANADA

phone: 506-364-2519
fax: 506-364-2505
email: jehrman-at-mta.ca
www: http://www.mta.ca/dmf

I before E except after C.
We live in a weird society!


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From: klivi-at-jhu.edu
Date: Tue, 21 Sep 2010 11:48:06 -0500
Subject: [Microscopy] Re: Objective lens in SEM?

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

Exactly! Microprobers have known this for a while and is very
instructive! Also the way to demonstrate why you should use normal
area scans for average EDS compositions (i.e., more time spent on the
flyback trace than on the area of interest)!
Ken

On Sep 21, 2010, at 12:19 PM, jehrman-at-mta.ca wrote:

}
}
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}
} If you're lucky enough to have a microprobe or SEM with an optical
} microscope, demonstrating magnification in an SEM is really slick. Put
} the scan on a piece of polished benitoite or other fluorescing
} mineral,
} and have the student look through the optical scope while playing with
} the magnification and scan speed. It's mind-blowing the first time you
} see it and drives home the 1-to-1 relationship between the scan on the
} specimen to the display scan. You can also see the flyback retrace and
} other things that have an effect on imaging and specimen
} contamination.
} Adjusting focus is cool too, which also helps to show what the
} "objective" lens does.
}
} Jim
}
} --
}
} James M. Ehrman
} Digital Microscopy Facility
} Mount Allison University
} 63B York St.
} Sackville, NB E4L 1G7
} CANADA
}
} phone: 506-364-2519
} fax: 506-364-2505
} email: jehrman-at-mta.ca
} www: http://www.mta.ca/dmf
}
} I before E except after C.
} We live in a weird society!
}
}
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|_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
Kenneth JT Livi, PhD
Director, The High-Resolution Analytical Electron Microbeam Facility
of the Integrated Imaging Center
Departments of Earth and Planetary Sciences and Biology
Olin Hall
3400 N Charles Street
Johns Hopkins University
Baltimore, Maryland 21218 USA
| | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| |
| :|









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From: protrain-at-emcourses.com
Date: Tue, 21 Sep 2010 13:13:02 -0500
Subject: [Microscopy] Objective lens in SEM?

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

Hi All

OK what is the definition of an objective lens? As light optics set the
standards for microscopy surely the definition from light optics is the one
that should be used?

"In an optical instrument, the objective is the optical element that gathers
light from the object being observed and focuses the light rays to produce a
real image." In the SEM is this the role of the electronics?

My original thought.

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com




-----Original Message-----
X-from: frah0010-at-umn.edu [mailto:frah0010-at-umn.edu]
Sent: 21 September 2010 16:12
To: protrain-at-emcourses.com

I believe that the term "objective lens" simply derives from the fact that
it is closest to the object (i.e., specimen) being observed. There isn't a
function tied to its nomenclature, only its physical placement. In that
sense, it should be perfectly acceptable to call the "final" lens in a SEM
or electron microprobe an objective lens.

Best,
Ellery

-----------------
Ellery Frahm
Manager & Principal Analyst, Electron Microprobe Lab
Senior Research Fellow, Department of Geology & Geophysics
University of Minnesota - Twin Cities
http://probelab.geo.umn.edu










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From: TindallR-at-missouri.edu
Date: Tue, 21 Sep 2010 13:16:45 -0500
Subject: [Microscopy] Objective lens

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

Having briefly followed this discussion, I think it is important to consider a highly relevant question: In these hyper-partisan times, is it possible for a lens to be truly objective at all?

Thoughtfully,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




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From: oshel1pe-at-cmich.edu
Date: Tue, 21 Sep 2010 13:31:34 -0500
Subject: [Microscopy] Re: Objective lens

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It's all relativistic.

Phil

} Having briefly followed this discussion, I think it is important to
} consider a highly relevant question: In these hyper-partisan times,
} is it possible for a lens to be truly objective at all?
}
} Thoughtfully,
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar:
} http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: jehrman-at-mta.ca
Date: Tue, 21 Sep 2010 13:39:43 -0500
Subject: [Microscopy] Re: Objective lens

Contents Retrieved from Microscopy Listserver Archives
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...An excellent topic for a FOCUS GROUP! ;-)

JME


--

James M. Ehrman
Digital Microscopy Facility
Mount Allison University
63B York St.
Sackville, NB E4L 1G7
CANADA

phone: 506-364-2519
fax: 506-364-2505
email: jehrman-at-mta.ca
www: http://www.mta.ca/dmf

I before E except after C.
We live in a weird society!




On 21/09/2010 3:17 PM, TindallR-at-missouri.edu wrote:
}
}
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} Having briefly followed this discussion, I think it is important to consider a highly relevant question: In these hyper-partisan times, is it possible for a lens to be truly objective at all?
}
} Thoughtfully,
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
}
}
}


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From: RossLM-at-missouri.edu
Date: Tue, 21 Sep 2010 13:47:16 -0500
Subject: [Microscopy] Re: Objective lens

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But since it's below the other lenses shouldn't it be the subjective lens?

Lou
--
Sr. Research Specialist
University of Missouri
Electron Microscopy Core Facility
W136 Veterinary Medicine Building
Columbia, MO 65211
573.882.4777, fax 573.884.2227
RossLM-at-missouri.edu
http://www.emc.missouri.edu/


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From: FMonson-at-wcupa.edu
Date: Tue, 21 Sep 2010 15:10:07 -0500
Subject: [Microscopy] Objective lens

Contents Retrieved from Microscopy Listserver Archives
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But aren't the optics all somewhat turned around in an SEM compared to a light microscope or a TEM. The illumination seems to come from the direction of the detector which usually has no particular focusing involved. The point of view seems to be from up the column. The focusing is done with the "objective" lens.

Frankly, I was happy with the idea that the objective lens is the one closest to the object.

BTW, I do really like the idea of demonstrating the concepts with benitoite. Now, if I could only do that using my chamberscope on the SEM.

Warren Straszheim

-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: Tuesday, September 21, 2010 1:14 PM
To: wesaia-at-iastate.edu

Given the state of 'everything' - which is mostly statistical, what is relevant is whether the 'cone' is even approximate! That, above all, differentiates it from the true 'objective' in the LM. There, we can distinguish between the virtual and real images.

Further, we can discount this discussion, because Missouri is a frontier state that is just a part of a large river delta. So there!

Frederick C. Monson, PhD
Technical Director, CMIRT
West Chester University
West Chester, PA, 19383
610-738-0437
New Web Site:           http://cmirt.wcupa.edu/index.html

New Scheduler:          http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl

Reads of the Month:
1. "The Liver as a Lymphoid Organ," Ian Nicholas Crispe, Annual Review of Immunology, Apr 2009, Vol. 27: 147-163. (Access Depends on subscription.)
2. "A Personal Journey of Discovery: Developing Technology and Changing Biology," Lee Hood, Annual Review of Analytic Chemistry, Vol. 1: 1-43.


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Tuesday, September 21, 2010 2:24 PM
To: Monson, Frederick

Having briefly followed this discussion, I think it is important to consider a highly relevant question: In these hyper-partisan times, is it possible for a lens to be truly objective at all?

Thoughtfully,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




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From: tomas.hrncir-at-tescan.cz
Date: Wed, 22 Sep 2010 02:21:07 -0500
Subject: [Microscopy] Re: Objective lens in SEM?

Contents Retrieved from Microscopy Listserver Archives
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Hi All,
unfortunately I do not know the article from "Microscopy Today", 1994 (by the way, Debby, could you provide the full reference?), but there is another reason for calling SEM final lens as an "objective".
While the beam crossover below condenser lens is generally projective (just virtual sometimes), crossover below the final lens is objective (always real) - we usually want to have the image as sharp as possible. Condensors are just projective lenses, that is quite big difference and I think you cannot call objective as "3rd condenser" because of this.
Anyway it is just a word, we can call SEM final lens e.g. "nose" and functionality will be the same (although I am afraid this term is not going to be generally accepted).
Have a nice day,
Tomas

--
Tomas Hrncir, Ph.D.
R&D - Physics
Tescan
Libusina trida 21
623 00 Brno - CZ
Phone: +420 547 130 468
Fax: +420 547 130 415
http://www.tescan.cz

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From: tgreco-at-marine.usf.edu
Date: Wed, 22 Sep 2010 06:42:54 -0500
Subject: [Microscopy] viaWWW: Pre-centered SEM filaments

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Email: tgreco-at-marine.usf.edu
Name: Tony Greco

Organization: College of Marine Science, University of South Florida

Title-Subject: [Filtered] Pre-centered SEM filaments

Message: I use pre-centered tungsten filaments in my SEM for
convenience but have noticed that while it allows one to quickly
replace a blown filament, the only control one has over filament
height is the flat copper washers used as spacers between the base
and grid cap. The filament height is preset for an average filament
current setting so the only way to get more current out of the gun
when doing high resolution work is to remove the spacers which I
frequently do. Usually this does not result in only a small increase
in filament current. As the filament ages the situation worsens and
it becomes increasingly difficult to perform high resolution work
since the filament current is so low and there is no way to increase
it.
Anyone else have this problem?
Tony Greco

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From: michael-at-shaffer.net
Date: Wed, 22 Sep 2010 08:40:46 -0500
Subject: [Microscopy] RE: [SPAM] viaWWW: Pre-centered SEM filaments

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

Tony writes ...


} Message: I use pre-centered tungsten filaments in my SEM for
} convenience but have noticed that while it allows one to quickly
} replace a blown filament, the only control one has over filament
} height is the flat copper washers used as spacers between the base
} and grid cap. The filament height is preset for an average filament
} current setting so the only way to get more current out of the gun
} when doing high resolution work is to remove the spacers which I
} frequently do. Usually this does not result in only a small increase
} in filament current. As the filament ages the situation worsens and
} it becomes increasingly difficult to perform high resolution work
} since the filament current is so low and there is no way to increase
} it.
} Anyone else have this problem?

Your description is consistent with my experience. That is, if you decrease
to tip-to-Wehnelt distance, you will need a higher filament temperature to
properly saturate the gun, which results in a slightly brighter
1st-crossover. However, I imaging this "increased brightness" will come
with diminishing returns at some point, while definitely shortening the
filament's lifetime. You could also try changing the "bias" for more
emission current -- what is your emission current now?

Why do you believe this behaviour is associated with "pre-centered"
filaments?
~~~~~~~~~~~~~~~~~~~~~~~~~
cheerios, michael shaffer  :o)
SEM-MLA Research Coordinator
INCO Innovation Centre
Memorial University
St. John's Newfoundland
SEM-MLA Calendar:
http://www.mun.ca/creait/maf/SEM-MLA/calendar.php





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From: maryflet-at-interchange.ubc.ca
Date: Wed, 22 Sep 2010 11:22:26 -0500
Subject: [Microscopy] viaWWW: Pre-centered SEM filaments

Contents Retrieved from Microscopy Listserver Archives
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Dear Tony,
If your interest is maintaining high-resolution performance, then decreasing
the filament-to-cap distance is one way to increase the filament emission
current. Another is to make sure the filament is fully saturated and check
the saturation and alignment frequently to make sure they are maximized. You
can buy pointed filaments that decrease the initial emission area on the
filament tip, thus decreasing your spot size. If you have control of the
bias, turn it down (or up, depending on model) to maximize emission current.
I have made my own pointed filaments by gently rubbing the new filament on
1200 grit paper, both sides of the tip, to decrease the size of the tip. All
these measures will decrease filament life, some drastically, although I
successfully used the self-pointed filament at 5 kV and it never burnt out.
Finally, some SEMs can be converted to LaB6 guns, which are both brighter
and longer-lasting than tungsten filaments. All these measures will cost
money, some more than others.
Good luck

Mary Fletcher
Electron Microscopist
Materials Eng. UBC
#309 - 6350 Stores Road
Vancouver, B.C. V6T 1Z4
Canada
Tel: 604-822-5648
Fax: 604-822-3619
email: maryflet-at-interchange.ubc.ca


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Email: tgreco-at-marine.usf.edu
Name: Tony Greco

Organization: College of Marine Science, University of South Florida

Title-Subject: [Filtered] Pre-centered SEM filaments

Message: I use pre-centered tungsten filaments in my SEM for
convenience but have noticed that while it allows one to quickly
replace a blown filament, the only control one has over filament
height is the flat copper washers used as spacers between the base
and grid cap. The filament height is preset for an average filament
current setting so the only way to get more current out of the gun
when doing high resolution work is to remove the spacers which I
frequently do. Usually this does not result in only a small increase
in filament current. As the filament ages the situation worsens and
it becomes increasingly difficult to perform high resolution work
since the filament current is so low and there is no way to increase
it.
Anyone else have this problem?
Tony Greco

Login Host: 131.247.136.116
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From: tgreco-at-marine.usf.edu
Date: Wed, 22 Sep 2010 11:35:02 -0500
Subject: [Microscopy] Pre-centered SEM filaments

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



Michael
With a new pre-centered SEM filament, the emission current is about 55uA at 15KV. The filament I presently have has about 24 hours on it and the emission current has already dropped to 46uA. The bias setting on the software slider is set at 0 which gives the maximum emission current. So you see my predicament. There is no way with these pre-centered filament caps for me to increase the emission current once the copper spacers are removed and the minimum bias is selected.

Tony

Tony writes ...



} } Message: I use pre-centered tungsten filaments in my SEM for
} } convenience but have noticed that while it allows one to quickly
} } replace a blown filament, the only control one has over filament
} } height is the flat copper washers used as spacers between the base
} } and grid cap. The filament height is preset for an average filament
} } current setting so the only way to get more current out of the gun
} } when doing high resolution work is to remove the spacers which I
} } frequently do. Usually this does not result in only a small increase
} } in filament current. As the filament ages the situation worsens and
} } it becomes increasingly difficult to perform high resolution work
} } since the filament current is so low and there is no way to increase
} } it.
} } Anyone else have this problem?
Your description is consistent with my experience. That is, if you decrease
to tip-to-Wehnelt distance, you will need a higher filament temperature to
properly saturate the gun, which results in a slightly brighter
1st-crossover. However, I imaging this "increased brightness" will come
with diminishing returns at some point, while definitely shortening the
filament's lifetime. You could also try changing the "bias" for more
emission current -- what is your emission current now?

Why do you believe this behaviour is associated with "pre-centered"
filaments?
~~~~~~~~~~~~~~~~~~~~~~~~~
cheerios, michael shaffer :o)
SEM-MLA Research Coordinator
INCO Innovation Centre
Memorial University
St. John's Newfoundland
SEM-MLA Calendar:
http://www.mun.ca/creait/maf/SEM-MLA/calendar.php



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From: FMonson-at-wcupa.edu
Date: Wed, 22 Sep 2010 13:08:01 -0500
Subject: [Microscopy] Objective lens

Contents Retrieved from Microscopy Listserver Archives
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Ah! Late in the day, he strikes from fatigue and disorientation - trying to be humorous and missing the point altogether. I have driven thru Missouri, and it is not a marsh, but a rather attractive part of the middle of our fine country. I like their football, and especially, their microscopy.

I am sorry. I should have been more circumspect and said only what I meant to say.

Thus, here is what I think, if anyone will forebear to read any more of my stuff.

There is one feature of the 'objective/projector/etc' component in the SEM that separates it completely from the 'objective' in the LM. In fact, there is no part of the column in the SEM that does the job of the LM objective, unless one speaks of the LM 'objective' in an inverted LM construct. In other words, the gun, the column, and the projector, in the SEM, do that which is done in the LM by the source of illumination and the so-called condenser.

If there is a component in the SEM that performs the same function as the objective in the LM, it is the detector.

As to magnification, which, I admit, may appear to confuse my assertion - unless one recognizes that the resolution of the LM is purchased by the construction/design of both the illuminator complex, including the condenser, and the 'objective' which is then positioned - by design - to collect the information 'projected' by the illumination passing thru the specimen.

Finally as to my introduction of the detector, there is the hazard that one might cry out that in the Confocal Laser Scanning Microscope, the objective plays the part of the condenser as well as the collector. I think, however,that nothing substantially changes my assertions concerning the comparisons of SEM to LM.

Finally, in the LM, we have all grown up learning to use it in spite of the fact that our visual system is forced to deal with a majority of out of focus information - the normal condition. The CLSM teaches us that in all that we perceive, there is very little that is in focus. The SEM provides us, like the astronomer and the atom smasher, the opportunity to pick among the various emissions and reflections, those parts which we wish to investigate at any given moment. The CLSM, the SEM, the TEM, and all of the other abiological 'imaging-by-detector' systems, provide us with the means to investigate the inner universe with bracketed perception. Much, much better.

Cheers,

Fred Monson


Frederick C. Monson, PhD
Technical Director, CMIRT
West Chester University
West Chester, PA, 19383
610-738-0437
New Web Site:           http://cmirt.wcupa.edu/index.html

New Scheduler:          http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl

Reads of the Month:
1. "The Liver as a Lymphoid Organ," Ian Nicholas Crispe, Annual Review of Immunology, Apr 2009, Vol. 27: 147-163. (Access Depends on subscription.)
2. "A Personal Journey of Discovery: Developing Technology and Changing Biology," Lee Hood, Annual Review of Analytic Chemistry, Vol. 1: 1-43.

-----Original Message-----
X-from: FMonson-at-wcupa.edu [mailto:FMonson-at-wcupa.edu]
Sent: Tuesday, September 21, 2010 4:21 PM
To: Monson, Frederick

Given the state of 'everything' - which is mostly statistical, what is relevant is whether the 'cone' is even approximate! That, above all, differentiates it from the true 'objective' in the LM. There, we can distinguish between the virtual and real images.

Further, we can discount this discussion, because Missouri is a frontier state that is just a part of a large river delta. So there!

Frederick C. Monson, PhD
Technical Director, CMIRT
West Chester University
West Chester, PA, 19383
610-738-0437
New Web Site:           http://cmirt.wcupa.edu/index.html

New Scheduler:          http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl

Reads of the Month:
1. "The Liver as a Lymphoid Organ," Ian Nicholas Crispe, Annual Review of Immunology, Apr 2009, Vol. 27: 147-163. (Access Depends on subscription.)
2. "A Personal Journey of Discovery: Developing Technology and Changing Biology," Lee Hood, Annual Review of Analytic Chemistry, Vol. 1: 1-43.


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Tuesday, September 21, 2010 2:24 PM
To: Monson, Frederick

Having briefly followed this discussion, I think it is important to consider a highly relevant question: In these hyper-partisan times, is it possible for a lens to be truly objective at all?

Thoughtfully,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




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From: bigelow-at-umich.edu
Date: Wed, 22 Sep 2010 14:16:10 -0500
Subject: [Microscopy] RE: SEM lens

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

Listers:

You are all correct to object to the practice of calling the second
lens in an SEM an objective lens. HOWEVER, I made the same complaint
some forty years ago, and it didn't do any good then, so I doubt that
doing so now will change anything. The terminology is so deeply
ingrained in common usage,especially in some prominent texts, that it
is very unlikely to be abandoned.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From: John.Mardinly-at-wdc.com
Date: Wed, 22 Sep 2010 15:19:58 -0500
Subject: [Microscopy] RE: SEM lens

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

Just out of curiosity, I looked through the book by C.W. Oatley to see what
the man generally credited with being one of the principle inventors of the
SEM called the final lens. He never used the phrase 'objective lens' in his
book. So, how DID the final lens begin to become referred to as the
'Objective Lens'?

John Mardinly
Western Digital

-----Original Message-----
X-from: bigelow-at-umich.edu [mailto:bigelow-at-umich.edu]
Sent: Wednesday, September 22, 2010 12:25 PM
To: John Mardinly

Listers:

You are all correct to object to the practice of calling the second
lens in an SEM an objective lens. HOWEVER, I made the same complaint
some forty years ago, and it didn't do any good then, so I doubt that
doing so now will change anything. The terminology is so deeply
ingrained in common usage,especially in some prominent texts, that it
is very unlikely to be abandoned.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From: frah0010-at-umn.edu
Date: Wed, 22 Sep 2010 15:30:37 -0500
Subject: [Microscopy] SEM by any other name

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

If we object (no pun intended) to the "final lens" in a SEM being called an "objective lens," by that same logic (i.e., using optical microscopy and the functions of its components as the gold standard for all subsequent terminology), then should we not also protest the very name of the scanning electron microscope? Is a SEM really a "microscope" then? After all, the mechanism of image formation is completely different in a SEM versus a TEM or visible-light microscope. If a "microscope" requires image formation using lenses and either transmitted or reflected illumination in an optical system that forms a real image, then a SEM is not a microscope. Instead, it would be a "signal reconstructor" or something like that. The same goes for AFM and a dozen other techniques. Fortunately, language can be flexible, so we are free to define a "microscope" as an instrument that makes small things look bigger in a variety of possible ways -- since "microscope" comes from the Greek words for "to look" and "small," that should be satisfying.

Best,
Ellery

-----------------
Ellery Frahm
Manager & Principal Analyst, Electron Microprobe Lab
Senior Research Fellow, Department of Geology & Geophysics
University of Minnesota - Twin Cities
http://probelab.geo.umn.edu






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From: oshel1pe-at-cmich.edu
Date: Wed, 22 Sep 2010 15:48:44 -0500
Subject: [Microscopy] Re: SEM lens

Contents Retrieved from Microscopy Listserver Archives
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Good question.
My guess is uncritical analogy with the objective lens of the TEM.
Who did first use "objective lens" for the final lens of a SEM?

Phil

} Just out of curiosity, I looked through the book by C.W. Oatley to see what
} the man generally credited with being one of the principle inventors of the
} SEM called the final lens. He never used the phrase 'objective lens' in his
} book. So, how DID the final lens begin to become referred to as the
} 'Objective Lens'?
}
} John Mardinly
} Western Digital
}
} -----Original Message-----
} X-from: bigelow-at-umich.edu [mailto:bigelow-at-umich.edu]
} Sent: Wednesday, September 22, 2010 12:25 PM
} To: John Mardinly
} Subject: [Microscopy] RE: SEM lens
}
} Listers:
}
} You are all correct to object to the practice of calling the second
} lens in an SEM an objective lens. HOWEVER, I made the same complaint
} some forty years ago, and it didn't do any good then, so I doubt that
} doing so now will change anything. The terminology is so deeply
} ingrained in common usage,especially in some prominent texts, that it
} is very unlikely to be abandoned.
} --
} Wilbur C. Bigelow, Professor Emeritus
} Materials Sci. & Engr., Univ. of Michigan
} Ann Arbor, Michigan 48109-2136
} e-mail: bigelow-at-umich.edu;
} Fx:734-763-4788; Ph:734-975-0858
} Address mail to: 2911 Whittier Court
} Ann Arbor, MI 48104-6731

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: dsherman-at-purdue.edu
Date: Wed, 22 Sep 2010 15:55:04 -0500
Subject: [Microscopy] Objective lens in SEM?

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

To get back to the application of the Abbe Equation to lenses in the SEM,
the Rayleigh Criteria gives a value for the minimum resolvable detail, a
process that is limited by effects of diffraction. An ideal lens takes each
object point and represents it as an exact point in the image. A real lens
takes each of the object points and spreads them out into a circular disk
(airy disk) in the image plane whose diameter depends on the angular
aperture of the lens. The Rayleigh Criteria predicts the resolution based
on: d = 0.612 x wavelength/refractive index of media x sin of the half angle
of the lens aperture (alpha) where d is the point-to-point distance of two
objects that can be recognized as partially separated and thus distinct
objects. This exact equation was then applied by Abbe to lenses. Now,
although the Abbe Equation holds very well for optical lenses, it does not
hold nearly as well to electronmagnetic lenses.

However, can we agree that it still gives valuable information as to trend
of what is happening with an electromagnetic lens? For instance. Both the
Rayleigh Criteria and Abbe Equation predict that the wavelength has a direct
relationship to resolution (shorter wavelength = smaller d or point-to-point
distance= better resolution).

In the case of SEM lenses, we adjust lens strength for each lens to produce
smaller spot sizes (airy disks?) to improve resolution potential on the
sample. Increased lens strength will have greater effect on the paths of the
electrons traveling through that lens, acting to move the paths in a spiral
fashion toward the optical axis of the lens. This culminates in a cross-over
on the optical axis (focal point) and in turn results in a smaller spot size
at that point as well as a larger half angle of the lens aperture (alpha).
Thus the sin of alpha (n being a value of 1 in a vacuum) has an indirect
relationship to d. When sin alpha increases, d decreases and potential
resolution increases.

Thus the Abbe equation, or the Rayleigh criteria applied to lenses can still
predict the effect of wavelength, lens strength and thus their effect on
aperture angle as measured from the merging beam above its crossover. This
in turn therefore can be used as a measure of resolution, not in actual
numerical value but in order to predict anticipated results given other
factors remaining constant (such as sample type).

In the case of the final lens (or objective lens, etc) in an SEM, it is
correct that no real image is formed by the passage of the electron beam
through the object such as in a TEM or a light microscope. However,
potential resolution still can be predicted by the Abbe equation. The beam
x-over must be at the level of the sample for the sample to be in focus.
However, shortening the distance between the sample and the bottom of the
lens (working distance) results in a larger alpha, smaller spot size and
better potential resolution.

Where things start to fall apart is in predicting the effect of electrons
having shorter wavelength (as a result of higher kV) in an SEM. Ideally the
shorter wavelength would provide greater resolution and this is so up to a
point. Then you get the effects of the spread of the beam within the sample
(interactive volume overlap as well as penetration into the sample) and the
more energetic electrons having shorter wavelengths may actually aggravate
the problem and resolution of surface detail becomes less. Another problem
is depth of focus as this also degrades when working distance is shorter so
final image may not appear as sharp if topography is such that all parts are
not in focus.

Comments?

Debby

Original article reference: Ivan Roth. Does a Scanning Electron Microscope
have an Objective lens?. 1994 (sorry, Vol 94 but don't know
number...probably #2-4 and no page numbers were used at that point.)
--
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy/


} From: "tomas.hrncir-at-tescan.cz" {tomas.hrncir-at-tescan.cz}
} Reply-To: "tomas.hrncir-at-tescan.cz" {tomas.hrncir-at-tescan.cz}
} Date: Wed, 22 Sep 2010 03:23:31 -0400
} To: Debby Sherman {dsherman-at-purdue.edu}
} Subject: [Microscopy] Re: Objective lens in SEM?
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} Hi All,
} unfortunately I do not know the article from "Microscopy Today", 1994 (by the
} way, Debby, could you provide the full reference?), but there is another
} reason for calling SEM final lens as an "objective".
} While the beam crossover below condenser lens is generally projective (just
} virtual sometimes), crossover below the final lens is objective (always real)
} - we usually want to have the image as sharp as possible. Condensors are just
} projective lenses, that is quite big difference and I think you cannot call
} objective as "3rd condenser" because of this.
} Anyway it is just a word, we can call SEM final lens e.g. "nose" and
} functionality will be the same (although I am afraid this term is not going to
} be generally accepted).
} Have a nice day,
} Tomas
}
} --
} Tomas Hrncir, Ph.D.
} R&D - Physics
} Tescan
} Libusina trida 21
} 623 00 Brno - CZ
} Phone: +420 547 130 468
} Fax: +420 547 130 415
} http://www.tescan.cz
}
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From: ute.frevert-at-nyumc.org
Date: Wed, 22 Sep 2010 16:40:02 -0500
Subject: [Microscopy] viaWWW: Move a TEM

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Email: ute.frevert-at-nyumc.org
Name: Ute

Organization: NYU School of Medicine

Title-Subject: [Filtered] Move a TEM

Message: My entire department is moving to a new building and we need
to relocate our 20 year-old Zeiss EM 910. I am thinking about asking
Zeiss to do the packing, moving and reinstallation for us - unless
there are other options, i.e. companies that are specialized in
moving such equipment. This is the first time I have to do something
like this. Any advice is highly appreciated.

Ute

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From: ahmad_ds-at-yahoo.com
Date: Thu, 23 Sep 2010 06:45:21 -0500
Subject: [Microscopy] viaWWW: Fischione Disk Grinder 160

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Email: wxufocus-at-gmail.com
Name: Jay He

Organization: Consulting

Title-Subject: [Filtered] Darkfield Kohler Illumination

Message: Hi, I'm recently working on the illumination system of a
custom made reflected light inspection microscope. The lab wanted to
add a coaxial lighting with both bright and darkfield capability.
Right now it uses external lighting.

I understand the Kohler system as a standard way of providing a
uniform bright field illumination with the lamp filament (or LED in
this case) in the rear focal plane of the objective. But for the
standard DF objectives (like the ones by Nikon or Olympus,) the
illumination light goes through the outside barrel of the objective,
which as far as I understand does not have any optical elements
except the final ring mirror.

So I hope this is a simple question, is there such a thing called
"Darkfield Kohler Illumination"? Isn't it most optimal to have
parallel light ray entering the out pathway of the objective to have
maximum brightness and uniform DF illumination?

Jay He

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From tim.mccarron-at-btinternet.com Thu Sep 23 05:21:17 2010
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Email: ahmad_ds-at-yahoo.com
Name: Ahmad Ashkaibi

Organization: Al-Balqa University

Title-Subject: [Filtered] Fischione Disk Grinder 160

Message: Hello everybody,

I've got a Fischione Disk Grinder of the model 160, for the
pre-thinning of TEM metallic specimens, but I have no idea how to
actually use it. It's not illustrated neither on their website nor in
the manual.

I mean it's a heavy stainless steel cylinder with a hole at the
bottom and a micrometer at the top.

could anybody please describe to me how to use it?

Thanks in advance.


Regards,

Ahmad Ashkaibi

EM Unit
Materials Engineering Department
Al-Blaqa Applied University


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From: oshel1pe-at-cmich.edu
Date: Thu, 23 Sep 2010 07:15:24 -0500
Subject: [Microscopy] Re: SEM by any other name

Contents Retrieved from Microscopy Listserver Archives
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Ummm ... yes and no.
First, I agree that it doesn't really matter if the final lens of a
SEM is called an objective lens. I don't because I prefer functional
definitions (what is the whatzit's function?), and the final lens of
a SEM is not involved in image formation. Unlike the objective lens
of a light microscope or a TEM.
But, other than a fun dispute over pizza and beer, does it really
matter? No - as long as we are all clear what lens we're discussing
when we're teaching, writing, etc.

As for "microscope", I agree completely. But, this is a functional
definition, so I'm biased.
I also agree with your comment about SEM *not* being like a TEM.
People get confused by the "EM" part. A SEM is much more similar to a
AFM than a TEM. Hence, my comment earlier about a taxonomy of
microscopes. (Just don't get me started on cladistics and transformed
cladistics ...)

Phil

} If we object (no pun intended) to the "final lens" in a SEM being
} called an "objective lens," by that same logic (i.e., using optical
} microscopy and the functions of its components as the gold standard
} for all subsequent terminology), then should we not also protest the
} very name of the scanning electron microscope? Is a SEM really a
} "microscope" then? After all, the mechanism of image formation is
} completely different in a SEM versus a TEM or visible-light
} microscope. If a "microscope" requires image formation using lenses
} and either transmitted or reflected illumination in an optical
} system that forms a real image, then a SEM is not a microscope.
} Instead, it would be a "signal reconstructor" or something like
} that. The same goes for AFM and a dozen other techniques.
} Fortunately, language can be flexible, so we are free to define a
} "microscope" as an instrument that makes small things look bigger in
} a variety of possible ways -- since "microscope" comes from the
} Greek!
} words for "to look" and "small," that should be satisfying.
}
} Best,
} Ellery
}
} -----------------
} Ellery Frahm
} Manager & Principal Analyst, Electron Microprobe Lab
} Senior Research Fellow, Department of Geology & Geophysics
} University of Minnesota - Twin Cities
} http://probelab.geo.umn.edu

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: oshel1pe-at-cmich.edu
Date: Thu, 23 Sep 2010 08:06:55 -0500
Subject: [Microscopy] Re: Objective lens in SEM?

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Debby,

Predicting the effects of higher beam energy is easy: higher energy =
greater penetration = greater beam spread in sample + greater
generation of SE II, SE III, & SEM IV noise secondary electrons =
less resolution. Ignoring for the moment greater beam damage in low-Z
- biological - specimens.
This is seen in Monte Carlo simulations of interaction volumes with
beam energy and in the classic image from Everhart et al. (1972
(Proc. 6th Int. Conf. on X-ray optics and Microanalysis, ed Shinoda
et al. U. Tokyo) - as seen in Sawyer & Grubb, Polymer Microscopy, 2nd
ed., pg. 27.
Wavelength as in Rayleigh & Abbe equations is irrelevant. Now, we're
dealing more with the SEM as a low-energy particle accelerator and
electon-atom collision cross sections.

Phil

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From: FMonson-at-wcupa.edu
Date: Thu, 23 Sep 2010 08:26:39 -0500
Subject: [Microscopy] SEM by any other name

Contents Retrieved from Microscopy Listserver Archives
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As you suggest, in the hands of those who do not know, the meaning of a word will surely be confused.
--------------------------------------------------------
http://www.merriam-webster.com/dictionary/microscope
--------------------------------------------------------
"Definition of MICROSCOPE
1
: an optical instrument consisting of a lens or combination of lenses for making enlarged images of minute objects; especially : compound microscope
2
: a non-optical instrument (as one using radiations other than light or using vibrations) for making enlarged images of minute objects {an acoustic microscope} "
--------------------------------------------------------
Also,

Dr. Bigelow has struck a chord, thus, calling
Memories of things that were galling.

Di(-)section vs. dis(-)section
Nuculus vs. nucleus
The 'Tragedy of the Commons' is about 'everyone else'!

Following Ellery's lead to avoid punning:

Everything that happens in a vacuum, runs out of gas, and perhaps this thread has done the same.

On the other hand, it is, almost, always instructive to kick terms and concepts around so that we understand, with greater clarity, the messages we send when we instruct and/or explain.

For example, is the SEM:
1. a microscope, when working to generate an elemental map (image),
2. and NOT, when it merely collects averaged elemental data from the same area on the specimen.

Too picky by a wide margin, perhaps, but must a microscope generate an image to keep its identity?

Terminal Cheers,

Fred

Frederick C. Monson, PhD
Technical Director, CMIRT
West Chester University
West Chester, PA, 19383
610-738-0437
New Web Site:           http://cmirt.wcupa.edu/index.html

New Scheduler:          http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl

Reads of the Month:
1. "The Liver as a Lymphoid Organ," Ian Nicholas Crispe, Annual Review of Immunology, Apr 2009, Vol. 27: 147-163. (Access Depends on subscription.)
2. "A Personal Journey of Discovery: Developing Technology and Changing Biology," Lee Hood, Annual Review of Analytic Chemistry, Vol. 1: 1-43.


-----Original Message-----
X-from: frah0010-at-umn.edu [mailto:frah0010-at-umn.edu]
Sent: Wednesday, September 22, 2010 4:37 PM
To: Monson, Frederick

If we object (no pun intended) to the "final lens" in a SEM being called an "objective lens," by that same logic (i.e., using optical microscopy and the functions of its components as the gold standard for all subsequent terminology), then should we not also protest the very name of the scanning electron microscope? Is a SEM really a "microscope" then? After all, the mechanism of image formation is completely different in a SEM versus a TEM or visible-light microscope. If a "microscope" requires image formation using lenses and either transmitted or reflected illumination in an optical system that forms a real image, then a SEM is not a microscope. Instead, it would be a "signal reconstructor" or something like that. The same goes for AFM and a dozen other techniques. Fortunately, language can be flexible, so we are free to define a "microscope" as an instrument that makes small things look bigger in a variety of possible ways -- since "microscope" comes from the Greek!
words for "to look" and "small," that should be satisfying.

Best,
Ellery

-----------------
Ellery Frahm
Manager & Principal Analyst, Electron Microprobe Lab
Senior Research Fellow, Department of Geology & Geophysics
University of Minnesota - Twin Cities
http://probelab.geo.umn.edu






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From: smithj-at-winthrop.edu
Date: Thu, 23 Sep 2010 09:29:24 -0500
Subject: [Microscopy] [TEM] Question about service response time

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

Folks
I'd appreciate hearing from owners of TEM's concerning manufacturer's
service response time--OFF-LIST, please.
And my question is about microscopes that are under manufacturer's
service contract--not third-party.
I've plenty of experience with third-party service, but my "n" with
major manufacturers of electron-optical equipment is rather small.
It is my opinion that one week is too long, but I'd like to hear from
others concerning your experience.
TIA
Julian

--
Julian P.S. Smith III
Director, Winthrop Microscopy Facility
Dept. of Biology
Winthrop University
520 Cherry Rd.
Rock Hill, SC 29733

803-323-2111 x6427 (vox)
803-323-3448 (fax)
803-524-2347 (cell)
Research Website www.birdnest.org/smithj
Personal Website www.rociada-east.net


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From: TindallR-at-missouri.edu
Date: Thu, 23 Sep 2010 09:59:00 -0500
Subject: [Microscopy] [TEM] Question about service response time

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

Julian,

X-from Hitachi, JEOL, and FEI (SEMs and TEMs) we normally have 24-48 hour or less response time for emergencies----meaning a scope being unusable. I have actually had response times of 2-3 hours. There have been a few occasions when we've waited longer, but only because there was absolutely nobody available. In most cases, though, if our primary service engineer is busy, they will call in someone else from another service area.

For non-emergencies, meaning the scope is usable, but not operating at 100%, we often need to wait longer. In general, the more serious the problem, the faster the response. We make an effort to work with our service providers to not make unreasonable demands for non-critical repairs, and they certainly are great at working with us.

Also, we often are able to fix things over the phone. In these cases, response time for a callback from a service engineer can be a few minutes to an hour or so.

All in all, we're very pleased with our manufacturers' service, which is why I'm copying this to the list. Credit where credit is due.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



-----Original Message-----
X-from: smithj-at-winthrop.edu [mailto:smithj-at-winthrop.edu]
Sent: Thursday, September 23, 2010 9:31 AM
To: Tindall, Randy D.

Folks
I'd appreciate hearing from owners of TEM's concerning manufacturer's
service response time--OFF-LIST, please.
And my question is about microscopes that are under manufacturer's
service contract--not third-party.
I've plenty of experience with third-party service, but my "n" with
major manufacturers of electron-optical equipment is rather small.
It is my opinion that one week is too long, but I'd like to hear from
others concerning your experience.
TIA
Julian

--
Julian P.S. Smith III
Director, Winthrop Microscopy Facility
Dept. of Biology
Winthrop University
520 Cherry Rd.
Rock Hill, SC 29733

803-323-2111 x6427 (vox)
803-323-3448 (fax)
803-524-2347 (cell)
Research Website www.birdnest.org/smithj
Personal Website www.rociada-east.net


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From: nicholls-at-uic.edu
Date: Thu, 23 Sep 2010 11:49:56 -0500
Subject: [Microscopy] [TEM] Question about service response time

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

Julian

Basically exactly the same experience as Randy reported from Hitachi & JEOL.

1-2 days for emergency response if the microscope is down. Longer if the
microscope is useable and there is not an engineer immediately available.
Both companies have service centers in Chicago which helps - i think the
fastest response I have had from JEOL is 30 minutes! The engineer was
already on campus at another microscope and was monitoring the vacuum after
a bake.

Alan

At 09:59 AM 9/23/2010, TindallR-at-missouri.edu wrote:



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

Alan W Nicholls, PhD
Interim Associate Director - RRC
Director of Research Service Facility (Electron Microscopy)
Research Resources Center - East (M/C 337)
Room 110 Science and Engineering South Building
The University of Illinois at Chicago
845 West Taylor St
Chicago, IL 60607-7058

Tel: 312 996 1227
Fax: 312 996 8091
Office: Room 110

Web site www.rrc.uic.edu


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From: FMonson-at-wcupa.edu
Date: Thu, 23 Sep 2010 13:08:12 -0500
Subject: [Microscopy] RE: SEM lens

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

John, that's the easiest question to answer.

Just like the case of the "Tragedy...", the most likely culprit would be a biologist who, not knowing better, thought the column looked like it was capped with a perfect analog of the 'objective' on his Leitz compound. After all, when I saw the inside of a SEM for the first time, I thought, "Boy, what a big objective!" It took a while, and, until 2002, I never spent much focused time with either TEM or SEM beyond the image/result.

Further, when I arrived early to my first demonstration of a scanning probe microscope, it took at least 10 minutes of embarrassed intellectual frying before I admitted - to myself - that I could not determine which of the devices on the table was 'IT". The one thought I remember with clarity was something like, "There is nothing there that looks like a microscope (though there was a dissecting scope)." The next thought was, "Well, the inventors of this 'scope' apparently didn't think it needed to look like an 'optical' microscope [as if - he now understands - electromagnetic lenses were not 'optical' too].

Anyway, in closing, the only kind of 'scientist' left as a less-likely candidate is a psychologist, and s/he is not likely to use such tools. Of course, I would avidly read an admission from a chemist.

Cheers, with apologies for us all,

Fred Monson

Frederick C. Monson, PhD
Technical Director, CMIRT
West Chester University
West Chester, PA, 19383
610-738-0437
New Web Site:           http://cmirt.wcupa.edu/index.html

New Scheduler:          http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl

Reads of the Month:
1. "The Liver as a Lymphoid Organ," Ian Nicholas Crispe, Annual Review of Immunology, Apr 2009, Vol. 27: 147-163. (Access Depends on subscription.)
2. "A Personal Journey of Discovery: Developing Technology and Changing Biology," Lee Hood, Annual Review of Analytic Chemistry, Vol. 1: 1-43.


-----Original Message-----
X-from: John.Mardinly-at-wdc.com [mailto:John.Mardinly-at-wdc.com]
Sent: Wednesday, September 22, 2010 4:30 PM
To: Monson, Frederick

Just out of curiosity, I looked through the book by C.W. Oatley to see what
the man generally credited with being one of the principle inventors of the
SEM called the final lens. He never used the phrase 'objective lens' in his
book. So, how DID the final lens begin to become referred to as the
'Objective Lens'?

John Mardinly
Western Digital

-----Original Message-----
X-from: bigelow-at-umich.edu [mailto:bigelow-at-umich.edu]
Sent: Wednesday, September 22, 2010 12:25 PM
To: John Mardinly

Listers:

You are all correct to object to the practice of calling the second
lens in an SEM an objective lens. HOWEVER, I made the same complaint
some forty years ago, and it didn't do any good then, so I doubt that
doing so now will change anything. The terminology is so deeply
ingrained in common usage,especially in some prominent texts, that it
is very unlikely to be abandoned.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From: SWalck-at-SouthBayTech.com
Date: Thu, 23 Sep 2010 13:26:48 -0500
Subject: [Microscopy] viaWWW: Fischione Disk Grinder 160

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

Ahmed,

The Fischione Model 160 hand grinder is similar in operation to other units
on the market. It uses a precision screw to push the sample holder out. If
you look in the hole of the grinder, you will see a little ball that is
spring loaded. This is positioned in a groove along the side of the
mounting stub for that unit. The mount can only be inserted one way. This
prevents the sample mount from turning. One complete turn of the micrometer
on top gives you 500 um linear motion along the axis of the grinder. The
sample is mounted on the sample mount with low temperature wax or with super
glue. The 160 has two types of mounts, solid and one that has a pyrex
insert in the center. I highly recommend the solid one for an application
where you are only thinning the sample down to a parallel sided sample with
a thickness between 60 and 100 um and do not need to see light through it.

There are two ways to use grinders like this. The first is to zero the stub
surface prior to mounting the sample. You can use a glass slide to find the
surface. Slide the glass slide across as you raise and lower the sample
stub (pushing the stub into the holder when you retract it). If you use a
drop of water to make a very thin film under the glass slide, you can use
that to find out when the stub pushes the glass slide above the surface of
the grinder because an air gap will occur. Then you simply move the dial
without turning the micrometer to zero the grinder. When you use this
method, you have to account for the thickness of the wax. With our
QuickStick(TM)-135 wax, the thickness is approximately 10 um thick if you
keep pressure on the sample while it cools. We have a spring loaded jig for
doing this, but you can also use a jig that applies pressure with a weight
on top. (see this link for an application note on the thickness and
uniformity of different types of waxes:
http://southbaytech.com/appnotes/72%20Comparing%20Wax%20Layer%20Thickness%20
after%20Mounting.pdf ) What you might want to do is set the zero of the
grinder for 10 um before mounting the sample. The other way to use the
micrometer is to realize that the micrometer is relatively very accurate.
So what you do there is to use a digital indicator stand to measure the
thickness of the sample while it is mounted on the sample stub. You could
use a digital indicator stand, such as our Model 102 indicator stand, which
allows you to measure in increments of 1 um. You zero on the stub and then
measure the thickness off the sample (+ wax) and then you can remove very
accurate amounts of material using the grinder's micrometer dial. This is
the method that I prefer using with every hand grinder that I have ever
owned. (I've owned and used them all in my career, including the Model
160.)

For using the hand grinder, you can use SiC abrasive papers, alumina or
diamond films. Use the most appropriate type for your material. You can use
a hand lapping tray, such as our model 180, for manual removal of material
or you can use a polishing station such as our model 910 or 920 lapping
machines. These can be set up for semi-automatic polishing with these types
of fixtures. For thinning TEM samples, I prefer to do hand grinding
manually a lapping tray. I can "feel" the sample thinning and can feel if
there are problems. For semiconducting materials, I like to use a hand
grinder using diamond films, usually a 3M 30 um (our part number
3MDFP08300-1) on the Model 180 lapping tray. You attach the plain backed
films on the glass surface using distilled water and squeezing out the water
between the film and the glass plate so there are no bubbles. The 3M films
are hydrophobic and the water used as the lubricant during grinding stays on
the film and does not go under it causing the film to release. The diamond
films are reusable many times and are easily cleaned with a paper towel.
For other materials, SiC papers work well. A 320 grit is equivalent to a 30
um size. You use water on the paper and you can hold it down by hand on the
flat surface of the glass while you use the grinding fixture.

You didn't mention your type of material that you are thinning. Here are
some notes about different types of samples and the use of hand polishers
that might be helpful to you.
-Do not put extra load on the fixture, i.e. don't push on it. Let
the weight of the grinder do the work. You are just moving it laterally in a
circular motion.
-Do not push the sample out when it is already in contact with the
abrasive with the weight of the fixture on the sample. If your material is
brittle, very small increments are better. These types of samples thin very
rapidly.
-Remember, the full weight of the fixture is on the sample. For
other types of samples, do not push the sample out too far from the bottom
of the fixture to cause it to excessively rock. The fixture will rock on
the sample as a pivot point and this causes the flat surface of the fixture
to start rounding, especially on the outside. Over time, the surface will
not be flat. If you listen to the sound of polishing, you can hear when the
surface of the sample is planar with the bottom of the fixture. You can
also feel it.
-When it is time to clean the sample and the bottom of the fixture,
do not hold the fixture upside down while it is wet. You don't want water
mixed with grinding debris to go up into the working mechanisms of the
grinders. Naturally, you also don't want water to be allowed on the top of
the fixtures and be allowed to go into the mechanism.

For completeness in this long-winded discourse, the SBT lapping fixtures do
not work exactly the same way. There is a piston in the center and the
micrometer allows the piston with the sample attached on the mounting block
to fall the specific distance set by the micrometer. When these fixtures
are used in this way, a selectable weight can be applied for the load.
These fixtures can also be used in the manner described above, but they are
set up slightly differently. The surface of the sample is set to fall below
the bottom of the fixture, just as they are above, and then the piston is
locked in place with a set screw on the side of the fixture. In this mode,
the full weight of the fixture is on the sample, just as the hand grinders
that use a positive screw motion.


Disclaimer: South Bay Technology manufactures and sells measurement
fixtures, lapping fixtures, lapping stations, and abrasive consumables for
polishing and thinning materials for microscopy applications.

-Scott

Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA 92673

eMail: SWalck-at-SouthBayTech.com
Phone: 949-492-2600
Toll Free: 1-800-728-2233 (USA)
Fax: 949-492-1499

www.SouthBayTech.com


-----Original Message-----
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Sent: Thursday, September 23, 2010 4:56 AM
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Email: ahmad_ds-at-yahoo.com
Name: Ahmad Ashkaibi

Organization: Al-Balqa University

Title-Subject: [Filtered] Fischione Disk Grinder 160

Message: Hello everybody,

I've got a Fischione Disk Grinder of the model 160, for the
pre-thinning of TEM metallic specimens, but I have no idea how to
actually use it. It's not illustrated neither on their website nor in
the manual.

I mean it's a heavy stainless steel cylinder with a hole at the
bottom and a micrometer at the top.

could anybody please describe to me how to use it?

Thanks in advance.


Regards,

Ahmad Ashkaibi

EM Unit
Materials Engineering Department
Al-Blaqa Applied University


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==============================Original Headers==============================
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From: germpore-at-sonic.net
Date: Thu, 23 Sep 2010 15:22:18 -0500
Subject: [Microscopy] Source for Abbe Test Plate, Diffraction Grating Slides, & Diatom Test Slides?

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

I'm looking for sources for 3 kinds of slides for a Microscopy class
in which I'm an instructional assistant:

1) Abbe Test Plate slide
2) Diffraction Grating slide
3) Diatom Test slide

I know Klaus Kemp makes custom diatom test slides, but as I remember,
they're weakly sealed and hard not to destroy with oil immersion. Any
other sources?

The other two kinds of slides I haven't for sale anywhere, though I do
remember using them in a microscopy class I took some years back with
a lot of old equipment.

==============================Original Headers==============================
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From: mjbehr-at-dow.com
Date: Mon, 27 Sep 2010 10:40:38 -0500
Subject: [Microscopy] viaWWW: Microtomy of hygroscopic materials

Contents Retrieved from Microscopy Listserver Archives
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Email: mjbehr-at-dow.com
Name: Michael

Organization: Dow Chemical

Title-Subject: [Filtered] Microtomy of hygroscopic materials

Message: I am trying to section hygroscopic material, so I can not
use water. Any suggestions for liquid I can use for this?
I have tried ethanol, however, the surface tension does not seem to
be high enough, as the sections do not slide off the knife, or if
they do, they are crumpled and slide beneath the liquid surface.

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From: PhillipsT-at-missouri.edu
Date: Mon, 27 Sep 2010 11:21:16 -0500
Subject: [Microscopy] viaWWW: Microtomy of hygroscopic materials

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

Glycerol has been used in the past. I have tried it and it is a messy job. I believe RL Ornberg was the one who originated the approach with some work he did with Tom Reese. I don't have the original papers any more but my thesis lists "Ornberg, R.L. & Reese, T.S. (1980) A freeze-substitution method for localizing divalent cations: examples from secretory systems. Fedn. Proc. 39(10):2802-2808" which I think is the correct reference.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
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Email: mjbehr-at-dow.com
Name: Michael

Organization: Dow Chemical

Title-Subject: [Filtered] Microtomy of hygroscopic materials

Message: I am trying to section hygroscopic material, so I can not
use water. Any suggestions for liquid I can use for this?
I have tried ethanol, however, the surface tension does not seem to
be high enough, as the sections do not slide off the knife, or if
they do, they are crumpled and slide beneath the liquid surface.

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From: DusevichV-at-umkc.edu
Date: Mon, 27 Sep 2010 11:37:46 -0500
Subject: [Microscopy] RE: viaWWW: Microtomy of hygroscopic materials

Contents Retrieved from Microscopy Listserver Archives
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I have used ethylene glycol for cutting of resin embedded cell cultures (to preserve mineral on initial stages of mineralization). Results were much worse than with water, but still usable. Section should be dried for a very long time; I dried them for a week.

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

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} Message: I am trying to section hygroscopic material, so I can not
} use water. Any suggestions for liquid I can use for this?
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From: W.Muss-at-salk.at
Date: Mon, 27 Sep 2010 12:57:32 -0500
Subject: [Microscopy] Re: Microtomy of hygroscopic materials

Contents Retrieved from Microscopy Listserver Archives
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Good afternoon/good evening,
Dear Michael,

as to my knowledge, in some circumstances also DMSO has been used as trough liquid for ultrathin sectioning :

cf:
Histofluorescent labelling of catecholaminergic structures in rotifers (Aschelminthes)
J. Keshmirian and T. Nogrady
Histochemistry and Cell Biology
Volume 89, Number 2, 189-192, DOI: 10.1007/BF00489923
"....concentration of DMSO for sectioning was 4%, and markedly improved the quality of cutting..."

http://www.immunologie-labor.com/cellmarker_files/fach_ultracryo_1.pdf
KUHLMANN WD & VIRON A: Cross-Linked Albumin as Supporting Matrix in Ultrathin Cryo Microtomy
J. Ultrastructure Research 41, 385-394, 1972
"....DMSO and sectioned on 50% DMSO in the trough at about -50°C. Negative stain...."

http://www.diatomeknives.com/knives/cryo_knife.aspx (Disclaimer: no affiliation, no financial interest!)
Cryo Diamond Knife==} "....The triangular holder, suitable for dry sectioning, as well as the trough, for sectioning using fluids, (DMSO/water) are both made from a special copper-nickel alloy,...."

[Warning! DMSO readily penetrates skin and may carry other dissolved chemicals into the body. May cause eye, skin, and respiratory tract irritation. Combustible liquid and vapor. Hygroscopic (absorbs moisture from the air).
Target Organs: Central nervous system, eyes, skin ==} e.g. cp. http://fscimage.fishersci.com/msds/07770.htm ]

But: Dimethyl sulfoxide (DMSO) is a clear hygroscopic liquid; melting point 18 C; ... It is miscible with water; readily soluble in almost all organic solvents.

Just my 2 (Euro-)Cents
Best wishes and regards,

Wolfgang MUSS
EM-Lab,
Pathology Gen.Hosp. SALZBURG
AUSTRIA


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} Gesendet: Montag, 27. September 2010 17:46
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}
} I am trying to section hygroscopic material, so I can not use water.
} Any suggestions for liquid I can use for this?
} I have tried ethanol, however, the surface tension does not seem to be
} high enough, as the sections do not slide off the knife, or if they do,
} they are crumpled and slide beneath the liquid surface.
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From: thoward-at-unm.edu
Date: Mon, 27 Sep 2010 13:05:31 -0500
Subject: [Microscopy] Re: viaWWW: Microtomy of hygroscopic materials

Contents Retrieved from Microscopy Listserver Archives
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I haven't done this, but years ago a colleague had
embedded plant samples that were destined for EDAX & the
component of interest was water-soluble, so cutting with a
regular water-filled boat was out. She was able to cut the
sections dry, then pick up the (crumpled) sections & place
them on drops of ethylene glycol on slides. She then
stretched the sections with toluene vapor (the old stick
trick) & dried them down on a hotplate/slide warmer. My
notes say that she actually dried them with heat in the
presence of toluene vapor, but that would probably make
your safety department cry or have a collective nervous
breakdown, so maybe skip that part!

Tamara



On Mon, 27 Sep 2010 10:42:58 -0500
mjbehr-at-dow.com wrote:
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} I can not
} use water. Any suggestions for liquid I can use for
} this?
} I have tried ethanol, however, the surface tension does
} not seem to
} be high enough, as the sections do not slide off the
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} they do, they are crumpled and slide beneath the liquid
} surface.
}
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***************************
Tamara Howard
Cell Biology & Physiology
UNM-HSC
Albuquerque, NM
***************************


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From: oshel1pe-at-cmich.edu
Date: Tue, 28 Sep 2010 06:57:40 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist overseas shipping?

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One for the service companies and microscope suppliers.
Please be sure to respond directly to Mordaunt, and the list. I
suspect there are plenty of people on the microscopy list who would
like to know this as well.

} Date: Tue, 28 Sep 2010 04:04:50 -0700 (PDT)
} From: Richard Mordaunt {Sea_Brat2002-at-yahoo.com}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
} realname - Richard Mordaunt
} Email - Sea_Brat2002-at-yahoo.com
} ORGANIZATION - NEU
} EDUCATION - Undergraduate College
} LOCATION - Boston, MA, USA
} SUBJECT_OF_QUESTION - Packaging microscopes for overseas shipment
} QUESTION - A program recently closed at NEU and has a number of left
} over microscopes. Our plan is to donate about 6 of these to a
} non-profit. This requires shipment overseas. We are looking for a
} preparation & packaging method and materials. Microscopes are
} Leica DME model.
}
--
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From: ibarke2-at-uwo.ca
Date: Tue, 28 Sep 2010 08:27:27 -0500
Subject: [Microscopy] Canadian Source for 0-5 psig Pressure Gauges?

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Hello,
We require a 0-5 psi pressure gauge for our SEM, and we are having troubles locating a Canadian source for them.  We found one, but it was around 100 dollars. Does anyone have any ideas where we may locate a 0-5psi gauge (1/4" or 1/8" NPT threaded) within Canada?
Or, alternatively, does anyone have a 0-5psi gauge they would like to sell?
Please respond to ibarke2-at-uwo.ca
Thank you.


Ivan R. Barker
UWO Earth Sciences MSc
ZAPLab Geochronology Research Technician
T: 519-661-2111 ext. 88397
E: ibarke2-at-uwo.ca
 





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From: doc.vrdoljak-at-gmail.com
Date: Tue, 28 Sep 2010 15:40:17 -0500
Subject: [Microscopy] contract labs in SF bay area

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Hello,
Does anyone have a list of contract labs they can send me that are in
the San Francisco - Bay area, including silicon valley. We need labs
that can do SEM and EDX on recharge fees. This is for industrial
research projects.

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1, 30 -- Subject: contract labs in SF bay area
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From: nmedvitz-at-nephrocor.com
Date: Tue, 28 Sep 2010 19:52:41 -0500
Subject: [Microscopy] viaWWW: Job Opportunity in Orlando FL, renal pathology lab

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Email: nmedvitz-at-nephrocor.com
Name: Neil Medvitz

Organization: Bostwick Laboratories

Title-Subject: [Filtered] Job Opportunity in Orlando FL

Message: Below is a brief description of a EM job opportunity in a
renal pathology lab in Orlando Fl.

Position Summary

This position is in a renal pathology lab. Duties include grossing,
processing, embedding, cutting, staining, scoping, and imaging for
all aspects of the lab. Personnel must have the ability to be precise
and meticulous with strong attention to detail and the ability to
exercise initiative while working in a team environment, demonstrate
proven multitasking abilities and have a good verbal and written
communication skills.

Please feel free to contact me with any questions via email:
nmedvitz-at-nephrocor.com

Thanks,
Neil


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From: james.romanow-at-uconn.edu
Date: Tue, 28 Sep 2010 19:53:14 -0500
Subject: [Microscopy] viaWWW: Need Zeiss DSM9xx SEM SE detector assembly

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Email: james.romanow-at-uconn.edu
Name: Jim Romanow

Organization: The University of Connecticut

Title-Subject: [Filtered] Need Zeiss DSM9xx SEM SE detector assembly

Message: Hello,

Does anyone have a Zeiss DSM982 Gemini SE detector assembly available
for free or sale? The same basic part is also used on the Zeiss DSM
950/940 and 960 series SEMs. Condition of scintillator is
irrelevant. Please contact me off-line.

Thank you,

Jim


James S. Romanow
The University of Connecticut
Electron Microscopy Laboratory
BSP Building
Room G06, Unit 3242
91 North Eagleville Road
Storrs, CT 06269-3242

860 486-2914
james.romanow-at-uconn.edu


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From: samuel.connell-at-stjude.org
Date: Wed, 29 Sep 2010 17:35:36 -0500
Subject: [Microscopy] viaWWW: Director of Light Microscopy Position

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Email: samuel.connell-at-stjude.org
Name: Samuel Connell

Organization: St. Jude Children's Research Hospital

Title-Subject: [Filtered] Director of Light Microscopy Position

Message: Director of Light Microscopy
Cell and Tissue Imaging Center
St. Jude Childrenís Research Hospital
(Job Number 18238)

We announce an exciting opportunity for experts
in live cell imaging technology to direct a high
profile facility advancing research in cancer
cell biology. The Cell and Tissue Imaging Center
(CTIC) at St. Jude Childrenís Research Hospital
combines technology with teamwork to facilitate
studies that advance our understanding of
fundamental biological processes and our ability
to treat or prevent catastrophic diseases of
childhood. The CTIC is a centralized, highly
specialized shared resource for the science of
light microscopy. The staff consists of the
Director of Light Microscopy and three Imaging
Scientists. The CTIC has recently undergone a
$2.5M renovation, demonstrating a long-term
institutional commitment to provide
state-of-the-art imaging instrumentation and
facilities to the research community at St. Jude
Childrenís Research Hospital. The CTIC is
dedicated to helping to solve challenging
biological questions with the optimal acquisition
and analysis methodologies.

The Director of the Light Microscopy Facility is
responsible for managing the light microscopy
facility within the institutionís Cell and Tissue
Imaging Center (CTIC) and for collaborating with
faculty and staff on research projects. This
center includes equipment for confocal laser
scanning microscopy, multiphoton microscopy,
spinning disc confocal microscopy, spectral
imaging, fluorescence correlation spectroscopy,
TIRF, widefield and cell microinjection. Of
particular interest is instrumentation optimized
for single molecule techniques, such as PALM,
STORM and Fluorescence Correlation Spectroscopy.
The Director of the Light Microscopy Facility
coordinates experimental design for advanced
light microscopy; trains faculty and staff on
experimental design, light microscopy and data
processing and analysis; and maintains image
archives.

Requirements:

Bachelor's degree in an appropriate scientific
field plus 10 years of post-degree relevant and
productive work experience. OR Master's degree in
an appropriate scientific field plus 9 years of
post-degree relevant and productive work
experience. OR PhD degree in an appropriate
scientific field plus 5 years of post-degree
relevant and productive work experience.
Demonstrated excellence in the management of a
light microscopy shared resource is preferred.
Demonstrated progression in continuing education
in advanced light microscopy techniques is
required.

TO FIND OUT MORE ABOUT THIS POSTION AN TO APPLY GO TO www.stjude.org/jobs

St. Jude offers a positive working culture,
professional advancement and competitive
compensation.

St. Jude is an Equal Opportunity Employer and a Drug-Free Workplace.

Candidates receiving offers of employment will be
subject to pre-employment drug testing and
background checks.

--
Samuel A. Connell
Director of Light Microscopy
Cell & Tissue Imaging Center
St. Jude Children's Research Hospital
262 Danny Thomas Place
Memphis, TN 38105-3678
Office (901) 595-2536
Cell (901) 603-3162
samuel.connell-at-stjude.org

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From: oshel1pe-at-cmich.edu
Date: Thu, 30 Sep 2010 09:01:17 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist

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

} Below is the result of your form, submitted on Wednesday, September
} 29, 2010 at 08:03:19 PM.
}
} realname - Ruchi Malik
} Email - ruchi.malik-at-ndsu.edu
} EDUCATION - Graduate College
} QUESTION - Hi,
}
} I have a question regarding imaging of live human cell (epithelial)
} under TEM. In my experiment, I want to add nanoparticles (organic
} in nature) to the cells and see how they interact with each other.
} (localization).
} Can I use this method?
} 1) prepare suspension of live cells (in a buffer which contains
} glycerol) and add my nanoparticles to it.
} 2) place a drop of above suspension to a copper grid.
} 3) stain the grid with uranyl formate.
}
} I am aware that there are more complicated and time consuming
} procedures reported in literature, but the reason why I want to use
} the above method is because its less expensive. Do you think its
} durable?
}
} Thank you for your time and consideration
}
} Ruchi
--
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From: rcmoretz-at-gmail.com
Date: Thu, 30 Sep 2010 10:40:04 -0500
Subject: [Microscopy] Re: Fwd: Ask-A-Microscopist

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

Wet, live samples are incompatible with the TEM for several reasons:
1) The TEM is a high vacuum device that will not tolerate wet
samples; that is one of the main reasons for the more complicated &
time consuming methods for getting the sample into the microscope.
2) The TEM has a limited thickness of penetration for imaging. A
whole cell cannot be imaged, even when dry.
3) Contrast in the TEM on untreated samples is very low, and it would
be impossible to distinguish your nanoparticles from the cell and its
components.
4) Radiation from the electron beam would kill the cells in a very short time.

A cursory review of the literature would be useful.

Roger Moretz, Ph.D. (retired)
(I developed wet specimen sample holders in the 1960's and 70's, so am
conversant with the literature.)

On Thu, Sep 30, 2010 at 10:06 AM, {oshel1pe-at-cmich.edu} wrote:
}
}
}
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} } Below is the result of your form, submitted on Wednesday, September
} } 29, 2010 at 08:03:19 PM.
} }
} } realname - Ruchi Malik
} } Email - ruchi.malik-at-ndsu.edu
} } EDUCATION - Graduate College
} } QUESTION - Hi,
} }
} } I have a question regarding imaging of live human cell (epithelial)
} } under TEM. In my experiment, I want  to add nanoparticles (organic
} } in nature) to the cells and see how they interact with each other.
} } (localization).
} } Can I use this method?
} } 1) prepare suspension of live cells (in a buffer which contains
} } glycerol) and add my nanoparticles to it.
} } 2) place a drop of above suspension to a copper grid.
} } 3) stain the grid with uranyl formate.
} }
} } I am aware that there are more complicated and time consuming
} } procedures reported in literature, but the reason why I want to use
} } the above method is because its less expensive. Do you think its
} } durable?
} }
} } Thank you for your time and consideration
} }
} } Ruchi
} --
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From: Graham.Knott-at-epfl.ch
Date: Thu, 30 Sep 2010 12:00:53 -0500
Subject: [Microscopy] Safety requirements for osmium plasma coater

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

Dear Microscopists,

We have an osmium plasma coater that was installed a couple of years ago under the guidelines set by the school's safety requirements. This means that the closed system has to be installed in a fume hood (in which the osmium vials are stored), in a laboratory that is on negative pressure with the area around the fume hood curtained off. The operator must wear heavy protective clothing and also an independent breathing helmet. No one is allowed to enter the lab whilst the machine is in operation.

The consequence of all these requirements means that no one uses it, despite a lot of evidence suggesting that it would be useful for a number of projects. I consider these requirements a little over the top, and would like the safety requirements re-evaluated.

I would like to understand how other laboratories around the world are required to use their machines. If you have any information, I would be very grateful to hear from you.

Many thanks,
Graham Knott


----------------------------------
Graham Knott PhD, Senior Scientist, Head of Bio EM Facility, EPFL, Switzerland
Office: AI 0143: Laboratory: AI 0342: Phone +41 21 6931862:

==============================Original Headers==============================
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7, 18 -- Subject: Safety requirements for osmium plasma coater
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From: gregory.a.jerman-at-nasa.gov
Date: Thu, 30 Sep 2010 14:55:02 -0500
Subject: [Microscopy] Re: Safety requirements for osmium plasma coater

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

Our osmium plasma coater is located in a negative pressure room and operates
in a fume hood. Our osmium tetroxide ampoules are also stored in the hood
which runs 24 hours a day. We don't use any separate curtain. As long as
your hood is operating properly, and you use it properly, the constant air
flow into the hood keeps the likelihood of danger extremely low in the rest
of the room. We don't use heavy protective clothing or any kind of
breathing helmet. That seems a little over the top. As long as you have a
good pair of gloves and safety glasses, you shouldn't need any other
personal protective equipment. The osmium ampoules are only cracked open
inside the coater, so there is little concern for migration. Our vacuum
pump also has an activated charcoal filter on the outlet to catch any stray
osmium.

We only worry about three things. First is dropping and breaking an
ampoule. If it happens in the hood, no big deal. If it happens on the
floor outside the hood, just get out, close the door and the hood and
negative pressure in the room will vent the gas out. That is the whole point
of having a negative pressure room. Second is dealing with the broken
ampoule in the coater. When there is not enough osmium to operate the
coater, there will still be some trace amount left over, so you need to
evacuate the gas cell for a few hours to make sure it is all gone before
opening it up and removing the glass shards for disposal. The last is
handling and disposing of the vacuum pump oil and activated charcoal filter
properly since they will be contaminated.

Greg

Gregory A. Jerman
NASA MSFC
EM31 Materials Diagnostics Team
gregory.a.jerman-at-nasa.gov



} From: "Graham.Knott-at-epfl.ch" {Graham.Knott-at-epfl.ch}
} Reply-To: "Graham.Knott-at-epfl.ch" {Graham.Knott-at-epfl.ch}
} Date: Thu, 30 Sep 2010 12:05:38 -0500
} To: "Jerman, Gregory A. (MSFC-EM31)" {gregory.a.jerman-at-nasa.gov}
} Subject: [Microscopy] Safety requirements for osmium plasma coater
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear Microscopists,
}
} We have an osmium plasma coater that was installed a couple of years ago under
} the guidelines set by the school's safety requirements. This means that the
} closed system has to be installed in a fume hood (in which the osmium vials
} are stored), in a laboratory that is on negative pressure with the area around
} the fume hood curtained off. The operator must wear heavy protective clothing
} and also an independent breathing helmet. No one is allowed to enter the lab
} whilst the machine is in operation.
}
} The consequence of all these requirements means that no one uses it, despite a
} lot of evidence suggesting that it would be useful for a number of projects. I
} consider these requirements a little over the top, and would like the safety
} requirements re-evaluated.
}
} I would like to understand how other laboratories around the world are
} required to use their machines. If you have any information, I would be very
} grateful to hear from you.
}
} Many thanks,
} Graham Knott
}
}
} ----------------------------------
} Graham Knott PhD, Senior Scientist, Head of Bio EM Facility, EPFL, Switzerland
} Office: AI 0143: Laboratory: AI 0342: Phone +41 21 6931862:
}
} ==============================Original Headers==============================
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} 7, 18 -- Subject: Safety requirements for osmium plasma coater
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From: gw265-at-cam.ac.uk
Date: Thu, 30 Sep 2010 15:49:29 -0500
Subject: [Microscopy] cutting window on a Leica UCT ultramicrotome

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

Does anyone know why the setting of a cutting window on a Leica UCT might
not work? Or how to make it work? I'm using it at room temperature, i.e.
not for cryo work.

I'm not a technician, just a user, but the technician is as mystified as I
am, as to why it works sometimes and not others. I'm trying to set a
reasonably small cutting window, but not impossibly small - a distance
that works fine on other ultramicrotomes.

We're following the instructions - it's worked two of the last six times
I've tried to use it (spread over the course of a couple of years, with
intermittent other users in between who mostly use it for cryo-work)

thanks

Giselle

==============================Original Headers==============================
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From: jason.gillen-at-encorewire.com
Date: Thu, 30 Sep 2010 18:58:25 -0500
Subject: [Microscopy] viaWWW: SEM-EDS sample prep for metals and polymers wish list

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Email: jason.gillen-at-encorewire.com
Name: Jason Gillen

Title-Subject: [Filtered] SEM-EDS sample prep for metals and polymers wish list

Message: Anyone have a list of SEM-EDS sample prep accesories that
have proven useful for metals and polymer work. I will have the
normal set up of diamond saws, grinders/polishers, sputter coaters
but if there are other devices/materials that are nice to have for
proper sample prep for these materials I would appreciate any input.
I would like to purchase these now while my budget is still
breathing....Thanks

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From: oshel1pe-at-cmich.edu
Date: Fri, 1 Oct 2010 10:17:04 -0500
Subject: [Microscopy] Fwd: Ask-A-Microscopist

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The requester's name should always be at the top, before the
question, along with their affiliation and email address. It's part
of the form they fill out.
But, since I get these requests sent to me automatically by the web
server, there is no mechanism for any reply to go automatically to
the requester. Hence the footer I add. If it's too difficult to read
the entire message, I'll move the footer to the top.

Phil

} This was sent to the list and Phil Oshel who forwards the questions
} from the web form to the mail list.
}
} I had gotten into the habit of simply replying to such web-submitted
} questions and copying them to the entire list. However, now that
} Phil is often forwarding the questions, the initial requestor's name
} is buried further down and must be explicitly found and copied into
} an address field. A simple "reply" is not enough.
}
} I had given much the same response yesterday as a material scientist
} (without copying the list). It is nice to see someone else
} corroborate my points and with more authority.
}
} Warren
} ________________________________________
} From: rcmoretz-at-gmail.com [rcmoretz-at-gmail.com]
} Sent: Thursday, September 30, 2010 10:40 AM
} To: wesaia-at-iastate.edu
} Subject: [Microscopy] Re: Fwd: Ask-A-Microscopist
}
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--
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From: TindallR-at-missouri.edu
Date: Fri, 1 Oct 2010 10:27:04 -0500
Subject: [Microscopy] TEM: chloroform stretching of sections

Contents Retrieved from Microscopy Listserver Archives
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Dear Collective,

Maybe this is a dumb one, but it's Friday and I'll take the chance. When using chloroform to stretch resin sections in the knife boat, has anyone had the experience of having the sections stretch and tear more while under the TEM beam? Can the chloroform actually thin out and erode the sections significantly, as well as just relaxing them?

I'm working on a set of samples and the unstretched sections are displaying wrinkles, while the stretched ones seem to be tearing and developing holes. I'm hoping to go into the weekend with a peaceful mind, so I can enjoy the Blues festival: http://www.rootsnbluesnbbq.com/.

Thanks and

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: rschmitz-at-uwsp.edu
Date: Fri, 1 Oct 2010 10:28:45 -0500
Subject: [Microscopy] TEM: chloroform stretching of sections

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My students and I always stretch our sections with chloroform and we have horrible problems with the sections tearing and blowing up when exposed to the beam. The only way we can avoid it is by exposing the sections to the beam at the same level that we use to take a picture (yes we are still using film). We gradually increase brightness until we eventually able to go to crossover and focus. We use a EMBed/SPURS resin mixture (roughly 4parts/3parts). Also we are still using up our supply of the old SPURS formula.

Bob
Dr. Robert J. Schmitz
Associate Professor of Anatomical Sciences
Department of Biology
University of Wisconsin-Stevens Point
800 Reserve Street, 380 TNR Bld
Stevens Point, WI 54481
715-346-2420
Email: rschmitz-at-uwsp.edu
http://www.uwsp.edu/biology/faculty/rschmitz/index.html



________________________________
X-from: {TindallR-at-missouri.edu}
Reply-To: {TindallR-at-missouri.edu}

Dear Collective,

Maybe this is a dumb one, but it's Friday and I'll take the chance. When using chloroform to stretch resin sections in the knife boat, has anyone had the experience of having the sections stretch and tear more while under the TEM beam? Can the chloroform actually thin out and erode the sections significantly, as well as just relaxing them?

I'm working on a set of samples and the unstretched sections are displaying wrinkles, while the stretched ones seem to be tearing and developing holes. I'm hoping to go into the weekend with a peaceful mind, so I can enjoy the Blues festival: http://www.rootsnbluesnbbq.com/.

Thanks and

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: PhillipsT-at-missouri.edu
Date: Fri, 1 Oct 2010 10:54:34 -0500
Subject: [Microscopy] TEM: chloroform stretching of sections

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I don't know about your thin sections but your liver might stretch a tad after all that chloroform. I don't get why you don't use a heat pen. It is a lot safer.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Friday, October 01, 2010 10:28 AM
To: Phillips, Thomas E.

Dear Collective,

Maybe this is a dumb one, but it's Friday and I'll take the chance. When using chloroform to stretch resin sections in the knife boat, has anyone had the experience of having the sections stretch and tear more while under the TEM beam? Can the chloroform actually thin out and erode the sections significantly, as well as just relaxing them?

I'm working on a set of samples and the unstretched sections are displaying wrinkles, while the stretched ones seem to be tearing and developing holes. I'm hoping to go into the weekend with a peaceful mind, so I can enjoy the Blues festival: http://www.rootsnbluesnbbq.com/.

Thanks and

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: RRA-at-stowers.org
Date: Fri, 1 Oct 2010 10:54:39 -0500
Subject: [Microscopy] TEM: chloroform stretching of sections

Contents Retrieved from Microscopy Listserver Archives
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Dear Randy,

I don't use chloroform because I don't feel like passing out! But I do use xylene frequently. I use a harder resin than you do, and feel that would probably prevent the scenario you are describing. When I first started using it, I did seem to get the results you are talking about. Now I just use Epon hard formula. It seems better.

Thanks!
Rhonda Trimble

-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Friday, October 01, 2010 10:35 AM
To: Trimble, Rhonda

Dear Collective,

Maybe this is a dumb one, but it's Friday and I'll take the chance. When using chloroform to stretch resin sections in the knife boat, has anyone had the experience of having the sections stretch and tear more while under the TEM beam? Can the chloroform actually thin out and erode the sections significantly, as well as just relaxing them?

I'm working on a set of samples and the unstretched sections are displaying wrinkles, while the stretched ones seem to be tearing and developing holes. I'm hoping to go into the weekend with a peaceful mind, so I can enjoy the Blues festival: http://www.rootsnbluesnbbq.com/.

Thanks and

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: RRA-at-stowers.org
Date: Fri, 1 Oct 2010 11:17:07 -0500
Subject: [Microscopy] TEM: chloroform stretching of sections

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

I think I you are mis-informed about the "wish upon a star". Besides, I did order a heat pen from EMS, and the battery has to be brand new and strong for it to put out enough heat to stretch the sections. I talked with Vlad over at UMKC, and he does not like the expensive one, so I have held off on ordering one. I breathe a hell of a lot less xylene than I used to when working in histology labs for 20 years. That little bit is like nothing to me!

Thanks!
Rhonda Trimble


-----Original Message-----
X-from: Phillips, Thomas E. [mailto:PhillipsT-at-missouri.edu]
Sent: Friday, October 01, 2010 11:11 AM
To: Trimble, Rhonda

Dear Randy,

I don't use chloroform because I don't feel like passing out! But I do use xylene frequently. I use a harder resin than you do, and feel that would probably prevent the scenario you are describing. When I first started using it, I did seem to get the results you are talking about. Now I just use Epon hard formula. It seems better.

Thanks!
Rhonda Trimble

-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Friday, October 01, 2010 10:35 AM
To: Trimble, Rhonda

Dear Collective,

Maybe this is a dumb one, but it's Friday and I'll take the chance. When using chloroform to stretch resin sections in the knife boat, has anyone had the experience of having the sections stretch and tear more while under the TEM beam? Can the chloroform actually thin out and erode the sections significantly, as well as just relaxing them?

I'm working on a set of samples and the unstretched sections are displaying wrinkles, while the stretched ones seem to be tearing and developing holes. I'm hoping to go into the weekend with a peaceful mind, so I can enjoy the Blues festival: http://www.rootsnbluesnbbq.com/.

Thanks and

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: DusevichV-at-umkc.edu
Date: Fri, 1 Oct 2010 11:21:18 -0500
Subject: [Microscopy] TEM: chloroform stretching of sections

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

The heat pen does not work for my sections.
I still use chloroform and do not have problems under the beam.

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Manager
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

} -----Original Message-----
} From: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
} Sent: Friday, October 01, 2010 10:56 AM
} To: Dusevich, Vladimir
} Subject: [Microscopy] RE: TEM: chloroform stretching of sections
}
}
}
}
} -----------------------------------------------------------------------
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}
} I don't know about your thin sections but your liver might stretch a
} tad after all that chloroform. I don't get why you don't use a heat
} pen. It is a lot safer.
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://www.biology.missouri.edu/faculty/phillips.html
} http://www.biotech.missouri.edu/mcc/
}
}
} -----Original Message-----
} X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
} Sent: Friday, October 01, 2010 10:28 AM
} To: Phillips, Thomas E.
} Subject: [Microscopy] TEM: chloroform stretching of sections
}
}
}
}
} -----------------------------------------------------------------------
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} Dear Collective,
}
} Maybe this is a dumb one, but it's Friday and I'll take the chance.
} When using chloroform to stretch resin sections in the knife boat, has
} anyone had the experience of having the sections stretch and tear more
} while under the TEM beam? Can the chloroform actually thin out and
} erode the sections significantly, as well as just relaxing them?
}
} I'm working on a set of samples and the unstretched sections are
} displaying wrinkles, while the stretched ones seem to be tearing and
} developing holes. I'm hoping to go into the weekend with a peaceful
} mind, so I can enjoy the Blues festival:
} http://www.rootsnbluesnbbq.com/.
}
} Thanks and
}
} Cheers,
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar: http://biotech.rnet.missouri.edu/cgi-
} bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
}
}
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From: jb_sanderson-at-yahoo.com
Date: Fri, 1 Oct 2010 12:03:52 -0500
Subject: [Microscopy] Leitz periplan eyepiece wanted

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

Dear Colleagues,

I'm looking for a spare high-eyepoint Leitz Periplan 10x FoV 18
(part # 519815), for the older 23mm diameter RMS-standard eyepiece tubes.

This has got a 28mm x 0,75mm male thread (once you take off the rubber eyecup) that will screw into the from of the older Nikon Coolpix cameras.

cheers,
Jeremy

Jeremy Sanderson







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10, 32 -- Subject: Leitz periplan eyepiece wanted
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From: wtivol-at-verizon.net
Date: Fri, 1 Oct 2010 15:30:49 -0500
Subject: [Microscopy] Re: Fwd: Ask-A-Microscopist

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

Does anyone out there have any experience doing TEM/SEM on biofilms (~200uM
thick) grown on PC 3x1 inch slides with a 2uM clay on top? Customer would
like crossectional and topographic views to document prescence/distribution
of clay plates?

Fred Hayes
UC Davis


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3, 24 -- Subject: sample prep for clay/biofilm build
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From aaron-at-fasteninghouse.com Fri Oct 1 15:06:48 2010
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On Sep 30, 2010, at 7:13 AM, oshel1pe-at-cmich.edu wrote:

}
} }
} Below is the result of your form, submitted on Wednesday, September
} 29, 2010 at 08:03:19 PM.
}
} realname - Ruchi Malik
} Email - ruchi.malik-at-ndsu.edu
} EDUCATION - Graduate College
} QUESTION - Hi,
}
} I have a question regarding imaging of live human cell (epithelial)
} under TEM. In my experiment, I want to add nanoparticles (organic
} in nature) to the cells and see how they interact with each other.
} (localization).
} Can I use this method?
} 1) prepare suspension of live cells (in a buffer which contains
} glycerol) and add my nanoparticles to it.
} 2) place a drop of above suspension to a copper grid.
} 3) stain the grid with uranyl formate.
}
} I am aware that there are more complicated and time consuming
} procedures reported in literature, but the reason why I want to use
} the above method is because its less expensive. Do you think its
} durable?
}
} Thank you for your time and consideration
}
} Ruchi

Dear Ruchi,
It depends on the thickness of the cell and the high voltage of the
TEM. For your simple technique to work, the cell must be thin enough
for the beam to penetrate it without losing much energy; otherwise,
the lenses cannot form an image. It is sufficient that only the areas
of interest be thin; e.g., if the nanoparticles do not penetrate the
nucleus, you could still see them in the periphery of the cell. Also
bear in mind that the cell will be dried, if not during preparation,
then surely while being observed in the scope, so even a cell that is
a few um thick to begin with might still be thin enough when dried
out. I would make sure that the cell has indeed been dried before
placing it in the scope, since if the beam strikes a wet cell,
bubbling will drastically alter the structure. BTW, there is a step
4) to your procedure: rinse the grid to remove excess stain. If this
is done with a buffer that does not contain glycerol, the cell will
dry more quickly. Good luck.
Yours,
Bill


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From: gnonano-at-yahoo.com
Date: Sat, 2 Oct 2010 07:54:40 -0500
Subject: [Microscopy] viaWWW: high beam sensitive sample

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

Jason: I do the same sort of work and have found an ion beam
milling tool to be a great help. Especially if you have one that
can make a cross-section in less than 4 hours. The one I have
takes forever to make a section (and a dinky one at that) so I
use it for cleaning up soft material smear and layer
delineation. The ion mill can save you a lot of hair-pulling
when working with soft/ductile samples. Feel free to contact me
off-line for my opinions of the different vendor's tools.

On my wish list is an ultramicrotome for some of my polymer work.
One of our sister labs in Japan sent some polymer/copper laminate
stuff to an outside lab for cross-sectional analysis and they
faced the SEM section with an ultramicrotome. It was beautiful
and took a lot less time than me mechanically polishing.
=================================================================

Email: jason.gillen-at-encorewire.com
Name: Jason Gillen

Title-Subject: [Filtered] SEM-EDS sample prep for metals and
polymers wish list

Message: Anyone have a list of SEM-EDS sample prep accesories that
have proven useful for metals and polymer work. I will have the
normal set up of diamond saws, grinders/polishers, sputter coaters
but if there are other devices/materials that are nice to have for
proper sample prep for these materials I would appreciate any input.
I would like to purchase these now while my budget is still
breathing....Thanks

--
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Becky Holdford (r-holdford-at-ti.com)
214-567-0360
SC Packaging FA
Texas Instruments, Inc.
13536 N. Central Expressway MS 940
Dallas, TX 75243
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~


==============================Original Headers==============================
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7, 22 -- Subject: [Microscopy] viaWWW: SEM-EDS sample prep for metals and polymers
7, 22 -- wish list
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From apartridge-at-stageandsports.com Sat Oct 2 00:51:55 2010
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This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: gnonano-at-yahoo.com
Name: Pim

Organization: PhD

Title-Subject: [Filtered] high beam sensitive sample

Message: My sample is polymer blend (PS/EVOH) and EVOH phase is high
beam sensitive. What should I do to get rid of this problem?

This is my sample preparation ; Sample was microtomed to 50-70 nm
thickness and put in a copper grid. TEM with 120 KV was used to
examined.


Thank you very much

Login Host: 94.192.235.10
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From: bplowman-at-pacific.edu
Date: Sat, 2 Oct 2010 07:55:13 -0500
Subject: [Microscopy] viaWWW: Service contracts

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Email: bplowman-at-pacific.edu
Name: Barbara L. Plowman

Organization: University of the Pacific, Arthur A. Dugoni School of Dentistry

Title-Subject: [Filtered] Service contracts

Message: Our TEM was serviced once last year for preventative
maintenance and a new camera meter. We paid almost $10,000 for our
service contract. This is a large sum of money for an old scope that
does not see much wear and tear from too many users. Does anybody
have a more reasonable service contract for preventative maintenance
and/or limited service calls for a reliable, but old film TEM? I am
in the SF Bay area.

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From: Bplowman-at-pacific.edu
Date: Sat, 2 Oct 2010 07:55:36 -0500
Subject: [Microscopy] viaWWW: Chloroform and sections

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Email: Bplowman-at-pacific.edu
Name: Barbara L. Plowman

Organization: University of the Pacific Arthur a Dugoni School of Dentistry

Title-Subject: [Filtered] Chloroform and sections

Message: Yes, You can overstretch your sections with choloroform
making them more fragile under the electron beam. I try to use as
little chloroform as possible on my cotton swab. Then, I air it a
bit before waving it over the sections.


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From: smythen-at-musc.edu
Date: Sat, 2 Oct 2010 07:56:00 -0500
Subject: [Microscopy] viaWWW: Dalton's osmium

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Email: smythen-at-musc.edu
Name: Nancy Smythe

Organization: Medical University of South Carolina

Title-Subject: [Filtered] Dalton's osmium

Message: Does anyone have a recipe for Dalton's osmium? I am trying
to fix some spinal cord and this is what was suggested to me.

Thank you,

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From: Yeshiva.University.Careers-at-gmail.com
Date: Sat, 2 Oct 2010 07:56:31 -0500
Subject: [Microscopy] viaWWW: Job Opening - DIRECTOR OF LIGHT MICROSCOPY and IMAGE

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Email: Yeshiva.University.Careers-at-gmail.com
Name: Yeshiva University

Organization: Albert Einstein College of Medicine of Yeshiva University

Title-Subject: [Filtered] Job Opening - DIRECTOR
OF LIGHT MICROSCOPY and IMAGE ANALYSIS

Message: DIRECTOR OF LIGHT MICROSCOPY and Image Analysis



The Analytical Imaging Facility
(www.aecom.yu.edu/aif), a component of the
Gruss-Lipper Biophotonics Center (GLBPC) and a
core resource of the Albert Einstein College of
Medicine, is seeking a Director of Light
Microscopy and Image Analysis. We seek a
dynamic, motivated individual with expertise in
current techniques for fluorescence imaging of
cells and tissues. Candidates must have
demonstrated expertise in confocal microscopy,
FRAP, FRET and TIRF in fixed and live material,
facility to perform image analysis and to prepare
data for presentation and publication using
commercially available software such as MDS
MetaMorph, Perkin-Elmer Volocity, Bitplane Imaris
and Adobe Illustrator and Photoshop. Facility
with using and writing customized image process
routines in NIH ImageJ, MathWorks MATLAB and NI
LabVIEW will be useful.



Minimal requirements include a Masters Degree and
5 years experience in advanced fluorescence
microscopy imaging techniques preferably in a
core facility environment. The candidate must
have excellent written and verbal skills and be
able to interact productively with a diverse user
group.



The successful candidate will Direct the
operation of the light microscopy resources of
the AIF, the interface between the Biophotonics
Center and the AIF, supervise and train the staff
of light microscopy technicians, maintain the
facilityís inventory of light and laser confocal
microscopes, train users in microscope operation
and advanced imaging techniques, analysis of data
and preparation of figures for presentation and
publication. The successful candidate will
report directly to an executive committee
including the co-Directors of the GLBPC, and the
Scientific and Administrative Directors of the
Analytical Imaging Facility.



If you are looking for the opportunity to work in
a state of the art imaging facility with leading
biomedical scientists you are encouraged to
apply. This faculty position is a non-tenure
track full-time position with rank and salary
commensurate with qualifications. Applications
will be accepted until the position is filled.
To apply, a curriculum vitae, a cover letter
including statement of interests, salary
requirements and contact information for three
references should be e-mailed to Dr. John
Condeelis, Chair AIF Search Committee,
john.condeelis-at-einstein.yu.edu



Albert Einstein College of Medicine is an Equal
Opportunity/Affirmative Action Employer.


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From: bozzola-at-siu.edu
Date: Sat, 2 Oct 2010 08:45:40 -0500
Subject: [Microscopy] Re: viaWWW: Service contracts

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

This is a major issue facing many of us: contract or no contract. The
advantages of a contract are obvious and the lack of one is simply a
gamble. I do believe that older instruments, being of simpler
construction (and not using computers running Windoze software), are
less prone to breakdown. The older instruments could probably run
indefinitely, except for the lack of replacement parts.

By way of example, we have a 28 year old instrument that has been
under service contract since the beginning and a 15 year old
instrument that was never under service contract (other than the first
year warranty when it was purchased new). The 28 yr old is running as
well today as it did on day one, while the 15 yr old has been
non-functional for 2 yrs (due to the lack of a transformer - I am
told). Even if we had the 15 yr old on a contract, the part would
still not be available. So, in the latter case, a contract would have
been of doubtful value. The money we saved by not having coverage
would be enough to purchase a new instrument. Our plan now is to
search for the part (from a used instrument) and bring the instrument
back on line. In both instances, we made the right decision.

My personal preference is to always have coverage IF the money is
available. If not, then very carefully control the use of the
instrument and do the basic maintenance yourself. Call in the
manufacturer's technical support staff when you cannot fix it (or for
annual preventive maintenance). $10K per year for a TEM seems
reasonable, but you could half the cost by calling in service once a
year or as needed.

I hope others on this listserver will be able to provide some names of
service providers in your area. If not, stick with the manufacturer,
but on an as-needed basis.


--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730



} Email: bplowman-at-pacific.edu
} Name: Barbara L. Plowman
}
} Organization: University of the Pacific, Arthur A. Dugoni School of Dentistry
}
} Title-Subject: [Filtered] Service contracts
}
} Message: Our TEM was serviced once last year for preventative
} maintenance and a new camera meter. We paid almost $10,000 for our
} service contract.  This is a large sum of money for an old scope that
} does not see much wear and tear from too many users. Does anybody
} have a more reasonable service contract for preventative maintenance
} and/or limited service calls for a reliable, but old film TEM? I am
} in the SF Bay area.
}


==============================Original Headers==============================
10, 20 -- From bozzola-at-siu.edu Sat Oct 2 08:45:40 2010
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10, 20 -- Subject: Re: [Microscopy] viaWWW: Service contracts
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From: hyi-at-emory.edu
Date: Sat, 2 Oct 2010 09:47:52 -0400
Subject: [Microscopy] Re: viaWWW: Service contracts

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

About service on as-needed basis, be aware that all 4 major EM companies
charge a minimum of $250 per hour (data from 3 years ago) for service
(including travel time) if you do not have a contract. You also would have
to pay for all parts yourself. At this rate, $10K would only cover 3-4 days
of actual service work.

Hong


X-from: "bozzola-at-siu.edu" {bozzola-at-siu.edu}
Reply-To: "bozzola-at-siu.edu" {bozzola-at-siu.edu}

This is a major issue facing many of us: contract or no contract. The
advantages of a contract are obvious and the lack of one is simply a
gamble. I do believe that older instruments, being of simpler
construction (and not using computers running Windoze software), are
less prone to breakdown. The older instruments could probably run
indefinitely, except for the lack of replacement parts.

By way of example, we have a 28 year old instrument that has been
under service contract since the beginning and a 15 year old
instrument that was never under service contract (other than the first
year warranty when it was purchased new). The 28 yr old is running as
well today as it did on day one, while the 15 yr old has been
non-functional for 2 yrs (due to the lack of a transformer - I am
told). Even if we had the 15 yr old on a contract, the part would
still not be available. So, in the latter case, a contract would have
been of doubtful value. The money we saved by not having coverage
would be enough to purchase a new instrument. Our plan now is to
search for the part (from a used instrument) and bring the instrument
back on line. In both instances, we made the right decision.

My personal preference is to always have coverage IF the money is
available. If not, then very carefully control the use of the
instrument and do the basic maintenance yourself. Call in the
manufacturer's technical support staff when you cannot fix it (or for
annual preventive maintenance). $10K per year for a TEM seems
reasonable, but you could half the cost by calling in service once a
year or as needed.

I hope others on this listserver will be able to provide some names of
service providers in your area. If not, stick with the manufacturer,
but on an as-needed basis.


--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, ILÂ 62901
Phone: 618-453-3730



} Email: bplowman-at-pacific.edu
} Name: Barbara L. Plowman
}
} Organization: University of the Pacific, Arthur A. Dugoni School of Dentistry
}
} Title-Subject: [Filtered] Service contracts
}
} Message: Our TEM was serviced once last year for preventative
} maintenance and a new camera meter. We paid almost $10,000 for our
} service contract. Â This is a large sum of money for an old scope that
} does not see much wear and tear from too many users. Does anybody
} have a more reasonable service contract for preventative maintenance
} and/or limited service calls for a reliable, but old film TEM? I am
} in the SF Bay area.
}


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{AANLkTiksQ2ZM8zrN32-5mJJ6MtzQq8pc1fJ5L=-C6+iN-at-mail.gmail.com}
10, 20 -- Subject: Re: [Microscopy] viaWWW: Service contracts
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From: Frank_Karl-at-lincolnelectric.com
Date: Mon, 4 Oct 2010 06:13:58 -0500
Subject: [Microscopy] Re: Service contracts

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

I agree with Hong. Aging microscopes become more expensive to keep running. A high voltage problem can (and for us has) save you from repairs that would be far more expensive than the contract. Note also that contract clients are first in line for service engineers from most vendors.

John Minter
Eastman Kodak
On Oct 2, 2010, at 8:55 AM, bplowman-at-pacific.edu wrote:

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} Organization: University of the Pacific, Arthur A. Dugoni School of Dentistry
}
} Title-Subject: [Filtered] Service contracts
}
} Message: Our TEM was serviced once last year for preventative
} maintenance and a new camera meter. We paid almost $10,000 for our
} service contract. This is a large sum of money for an old scope that
} does not see much wear and tear from too many users. Does anybody
} have a more reasonable service contract for preventative maintenance
} and/or limited service calls for a reliable, but old film TEM? I am
} in the SF Bay area.
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} 6, 22 -- From zaluzec-at-microscopy.com Sat Oct 2 07:55:13 2010
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5, 23 -- From jrminter-at-rochester.rr.com Sat Oct 2 14:25:55 2010
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From mailingatwork-at-gmail.com Sat Oct 2 20:17:19 2010
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Message-ID: {005101cb62fc$ba1432db$b0d69aea-at-jhdgt}
Reply-To: "Movicont" {carlosduranavant-at-hotmail.com}

Service contract is, essentially, the health insurance for your instrument. You would "save" money if your instrument does not get sick, or simply get flu occasionally. You will be in big trouble if it needs a new heart.....

The turbo pump on my TEM went for self destruction a year ago. It would cost me } $35K, if I did not have the health insurance for him. He would also need to wait in the emergency room for a longer while.

X-from the managing point of view, if you run your facility in red, it would be "obvious" that you have a need. If, however, you "saved" a lot of money, it might be hard for you to ask "more" money from your institution. I know it doesn't make sense, but it seems to be the reality, for some of us, at least.

Zhaojie

Zhaojie Zhang, Ph.D.
Director, Jenkins Microscopy Facility
University of Wyoming



________________________________________
X-from: hyi-at-emory.edu [hyi-at-emory.edu]
Sent: Saturday, October 02, 2010 10:47 AM
To: Z.J. Zhang

About service on as-needed basis, be aware that all 4 major EM companies
charge a minimum of $250 per hour (data from 3 years ago) for service
(including travel time) if you do not have a contract. You also would have
to pay for all parts yourself. At this rate, $10K would only cover 3-4 days
of actual service work.

Hong


X-from: "bozzola-at-siu.edu" {bozzola-at-siu.edu}
Reply-To: "bozzola-at-siu.edu" {bozzola-at-siu.edu}

This is a major issue facing many of us: contract or no contract. The
advantages of a contract are obvious and the lack of one is simply a
gamble. I do believe that older instruments, being of simpler
construction (and not using computers running Windoze software), are
less prone to breakdown. The older instruments could probably run
indefinitely, except for the lack of replacement parts.

By way of example, we have a 28 year old instrument that has been
under service contract since the beginning and a 15 year old
instrument that was never under service contract (other than the first
year warranty when it was purchased new). The 28 yr old is running as
well today as it did on day one, while the 15 yr old has been
non-functional for 2 yrs (due to the lack of a transformer - I am
told). Even if we had the 15 yr old on a contract, the part would
still not be available. So, in the latter case, a contract would have
been of doubtful value. The money we saved by not having coverage
would be enough to purchase a new instrument. Our plan now is to
search for the part (from a used instrument) and bring the instrument
back on line. In both instances, we made the right decision.

My personal preference is to always have coverage IF the money is
available. If not, then very carefully control the use of the
instrument and do the basic maintenance yourself. Call in the
manufacturer's technical support staff when you cannot fix it (or for
annual preventive maintenance). $10K per year for a TEM seems
reasonable, but you could half the cost by calling in service once a
year or as needed.

I hope others on this listserver will be able to provide some names of
service providers in your area. If not, stick with the manufacturer,
but on an as-needed basis.


--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, ILÂ 62901
Phone: 618-453-3730



} Email: bplowman-at-pacific.edu
} Name: Barbara L. Plowman
}
} Organization: University of the Pacific, Arthur A. Dugoni School of Dentistry
}
} Title-Subject: [Filtered] Service contracts
}
} Message: Our TEM was serviced once last year for preventative
} maintenance and a new camera meter. We paid almost $10,000 for our
} service contract. Â This is a large sum of money for an old scope that
} does not see much wear and tear from too many users. Does anybody
} have a more reasonable service contract for preventative maintenance
} and/or limited service calls for a reliable, but old film TEM? I am
} in the SF Bay area.
}


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30, 27 -- From ZZhang-at-uwyo.edu Sat Oct 2 21:52:36 2010
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From assidai-at-sds.it Sun Oct 3 23:43:45 2010
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In my limited experience, organizations rarely, if ever set "saved" money
aside for a replacement scope. That soft savings goes to demonstrate
someone's management style and financial abilities. These same people are
seldom left holding the bag when the scope goes down for extended periods
or requires serious money to be made functional.

We all know the horror stories of scope or instrument company bought or
absorbed and now parts are not available or are no longer made for the
voltages or compatible with your mother board. Bad things happen to good,
well maintained scopes.

Upper management may yell at you over the cost of a service contract, but
let a scope go down and lose customers, business, grants and you will find
yourself in the cross hairs expanding why you didn't elucidate this
potential problem to them.

I side on the side of a service contract. At the very least, I too can
shrug and tell the boss, "The scope is so old, even the service guy can't
find parts..."


Enough typing without much more morning coffee........
Frank


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From: ALawrence-at-entomology.msstate.edu
Date: Mon, 4 Oct 2010 07:46:09 -0500
Subject: [Microscopy] Sputter Coater

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

We are hoping to purchase a new sputter coater that is capable of both metal sputtering and carbon evaporation.

Specially we've been looking at EMS 150T ES and would appreciate input good or bad from list members on this particular instrument. Comments on a comparable one by SPI or Ted Pella would be welcome as well.

Feel free to respond off line.

Thanks,
Amanda



==============================Original Headers==============================
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From: nizets2-at-yahoo.com
Date: Mon, 4 Oct 2010 08:58:10 -0500
Subject: [Microscopy] Re: Fwd: Ask-A-Microscopist

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

I wanted to react to the message of Bill.
Getting an image is a good thing. It is even better if the image makes sense. No
actually it is IMPORTANT that the image makes sense!
If I summarize the method here: The cells are air-dried without any preparation,
intoxicated with heavy metals (live!) and then irradiated with energetic
electrons. I ask you: what information do you want to obtain from such a
preparation?? If you want to know how cells look like when they are mistreated,
go further. But do not expect any meaningful results in terms of cellular
biology.

Regards,

Stephane


 
----- Original Message ----
X-from: "wtivol-at-verizon.net" {wtivol-at-verizon.net}
To: nizets2-at-yahoo.com
Sent: Fri, October 1, 2010 10:35:40 PM


On Sep 30, 2010, at 7:13 AM, oshel1pe-at-cmich.edu wrote:

}
} }
} Below is the result of your form, submitted on Wednesday, September
} 29, 2010 at 08:03:19 PM.
}
} realname - Ruchi Malik
} Email - ruchi.malik-at-ndsu.edu
} EDUCATION - Graduate College
} QUESTION - Hi,
}
} I have a question regarding imaging of live human cell (epithelial)
} under TEM. In my experiment, I want  to add nanoparticles (organic
} in nature) to the cells and see how they interact with each other.
} (localization).
} Can I use this method?
} 1) prepare suspension of live cells (in a buffer which contains
} glycerol) and add my nanoparticles to it.
} 2) place a drop of above suspension to a copper grid.
} 3) stain the grid with uranyl formate.
}
} I am aware that there are more complicated and time consuming
} procedures reported in literature, but the reason why I want to use
} the above method is because its less expensive. Do you think its
} durable?
}
} Thank you for your time and consideration
}
} Ruchi

Dear Ruchi,
    It depends on the thickness of the cell and the high voltage of the 
TEM.  For your simple technique to work, the cell must be thin enough 
for the beam to penetrate it without losing much energy; otherwise, 
the lenses cannot form an image.  It is sufficient that only the areas 
of interest be thin; e.g., if the nanoparticles do not penetrate the 
nucleus, you could still see them in the periphery of the cell.  Also 
bear in mind that the cell will be dried, if not during preparation, 
then surely while being observed in the scope, so even a cell that is 
a few um thick to begin with might still be thin enough when dried 
out.  I would make sure that the cell has indeed been dried before 
placing it in the scope, since if the beam strikes a wet cell, 
bubbling will drastically alter the structure.  BTW, there is a step 
4) to your procedure:  rinse the grid to remove excess stain.  If this 
is done with a buffer that does not contain glycerol, the cell will 
dry more quickly.  Good luck.
                        Yours,
                        Bill


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From: eschumacher-at-mccrone.com
Date: Mon, 4 Oct 2010 09:36:08 -0500
Subject: [Microscopy] Meeting Announcement: Midwest Microscopy and Microanalysis Society

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


I strongly agree with those who argue in favor of having major instruments on a manufacturer's service contract. (Note that there are independent service providers also who, by all accounts, do a very good job----I just don't have any personal experience with them.) Without a contract, you are at the mercy of luck and the service providers' hourly rates for service AND travel, plus expenses. Two or three days or this and you have already spent as much as a contract would cost. Except for one disastrous detour into the realm of insurance company maintenance coverage, we've had contracts on our scopes for forever.

Same goes for ancillary equipment, which often seems to be neglected. In my experience, it is much harder to get administrators to agree to contracts for smaller pieces of equipment (coaters, microtomes, etc.), but the running of a facility is as much dependent upon them as upon the major instruments. Obviously, if you can't prepare a specimen to get it into a $500,000 microscope because a $50,000 piece of equipment is down, what good is the scope? Service costs for support equipment can be just as high as for a major piece.

Case in point: we recently spent about $10,000 to repair a piece of equipment that had a service contract cost of about $5000. Costs included $350/hr for travel, which involved an engineer driving from quite a distance. Travel costs alone were ~$3000. The engineer was here for about two hours (and did an excellent job, incidentally), then had to order a part which I then later installed myself.

On the other hand, you can always take the chance and hope that nothing goes wrong.....

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From akotohj-at-francophonie.org Mon Oct 4 09:05:48 2010
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Greetings All,

The Midwest Microscopy and Microanalysis Society will hold its final meeting of 2010 on Friday, October 29th, at BP in Naperville, IL. The program theme is "In-Situ Transmission Electron Microscopy in Materials Science", and Robert Klie and Ke-Bin Low have put together an excellent line-up of speakers. Details and registration information can be found on our website under Meetings:

www.midwestmicroscopy.org

We look forward to seeing you there!

Elaine Schumacher
M3S Program Coordinator

*********************************************************************
Elaine F.Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com



*********************************************************************
This message and any attachments are solely for the
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disclosure, copying, use or distribution of the information
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From: kenconverse-at-qualityimages.biz
Date: Mon, 4 Oct 2010 09:43:33 -0500
Subject: [Microscopy] Service contracts

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

Can't say that I "inhaled". Isn't that what Clinton said? LOL

Thanks!
Rhonda Trimble


-----Original Message-----
X-from: Paula Sicurello [mailto:patpxs-at-gmail.com]
Sent: Saturday, October 02, 2010 11:40 AM
To: Trimble, Rhonda

Randy, et al,
Another aspect that I see as an independent, and I'm sure the manufacturers
see the same, is that a non-contract instrument often accumulates many small
problems over time. They are ignored, or lived with, because administrators
are loath to spend the money on a "minor" problem or even on "preventive
maintenance".

When the system eventually goes down, all they are willing to pay for is
enough to get the system so it is not "down". Rather a moving target, don't
you think?

Anyway, in these instances the customer is often not satisfied (user, not
administrator) and from a service engineer's standpoint it is VERY
unsatisfying. Our goal is to have things work correctly, reliably and to
specification. A service contract allows us to do what needs to be done
without having to watch the clock.

Sadly, if we do our job well, I often find that the microscopes end up
having not much more than their 2 preventive maintenance calls per year
after a few years because the major issues have been resolved and those,
often quick, preventive visits still can turn up and fix problems that the
user hasn't seen, yet. The bean counters look at that and say, "The
contract is costing much more than those 2 calls per year. Cancel it and
just schedule the preventive maintenance." Of course, the preventive
maintenance calls aren't scheduled because "it's running fine so we don't
need to spend the money." It's enough to make you pull your hair out.
Fortunately, mine still grows back.

DISCLAIMER: Quality Images is a third party (independent) service company
that has been servicing SEMs for over 29 years.

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Monday, October 04, 2010 10:02 AM
To: kenconverse-at-qualityimages.biz


I strongly agree with those who argue in favor of having major instruments
on a manufacturer's service contract. (Note that there are independent
service providers also who, by all accounts, do a very good job----I just
don't have any personal experience with them.) Without a contract, you are
at the mercy of luck and the service providers' hourly rates for service AND
travel, plus expenses. Two or three days or this and you have already spent
as much as a contract would cost. Except for one disastrous detour into the
realm of insurance company maintenance coverage, we've had contracts on our
scopes for forever.

Same goes for ancillary equipment, which often seems to be neglected. In my
experience, it is much harder to get administrators to agree to contracts
for smaller pieces of equipment (coaters, microtomes, etc.), but the running
of a facility is as much dependent upon them as upon the major instruments.
Obviously, if you can't prepare a specimen to get it into a $500,000
microscope because a $50,000 piece of equipment is down, what good is the
scope? Service costs for support equipment can be just as high as for a
major piece.

Case in point: we recently spent about $10,000 to repair a piece of
equipment that had a service contract cost of about $5000. Costs included
$350/hr for travel, which involved an engineer driving from quite a
distance. Travel costs alone were ~$3000. The engineer was here for about
two hours (and did an excellent job, incidentally), then had to order a part
which I then later installed myself.

On the other hand, you can always take the chance and hope that nothing goes
wrong.....

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar:
http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week
&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: TindallR-at-missouri.edu
Date: Mon, 4 Oct 2010 09:48:33 -0500
Subject: [Microscopy] viaWWW: high beam sensitive sample

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

My suggestion would be to put the sections on a carbon coated or formvar-carbon coated grid. This often helps dramatically when I have fragile sections---LR White immunolabeled ones, for example.

Another alternative is to keep the beam spread to avoid as much damage as possible and view the image on the monitor via a digital capture system, which can amplify an image that is hard to see on the fluorescent viewing screen. We have a user who requires very high resolution and magnifications, to the point that the viewing screen is pretty useless. He can capture decent images by using our digital camera and focusing with the Live Fast Fourier Transform function, using the shape of the central spot of the FFT image to tell when focus and stigmation are correct. The software we use is Digital Micrograph, but there must be other packages that will do similar things, provided that you have the digital capture system to begin with.

Your sample is thin enough that a lower KV beam would be fine, unless you need the resolution edge that higher KV's will give you. However, be aware that lower accelerating voltages can often cause more damage than high KV's.

Good luck.

Cheers,
Randy

-----Original Message-----
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Sent: Saturday, October 02, 2010 7:57 AM
To: Tindall, Randy D.

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Email: gnonano-at-yahoo.com
Name: Pim

Organization: PhD

Title-Subject: [Filtered] high beam sensitive sample

Message: My sample is polymer blend (PS/EVOH) and EVOH phase is high
beam sensitive. What should I do to get rid of this problem?

This is my sample preparation ; Sample was microtomed to 50-70 nm
thickness and put in a copper grid. TEM with 120 KV was used to
examined.


Thank you very much

Login Host: 94.192.235.10
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From: smithj-at-winthrop.edu
Date: Mon, 4 Oct 2010 10:09:38 -0500
Subject: [Microscopy] Re: Service Contracts

Contents Retrieved from Microscopy Listserver Archives
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Wow--I'd stay right where you are!
We pay nearly $17,000 to a major manufacturer, and our microscope was down for
a six-week period this summer (at the height of our student research
use) and has (as I type this) been down for the past twelve days, in
the middle of classes.
I think my experience with this particular
company may be unusually bad, but you should know that it could be a lot
more expensive than what you're presently paying.
Julian

On 10/2/10 9:11 AM, bplowman-at-pacific.edu wrote:
}
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}
} Email: bplowman-at-pacific.edu
} Name: Barbara L. Plowman
}
} Organization: University of the Pacific, Arthur A. Dugoni School of Dentistry
}
} Title-Subject: [Filtered] Service contracts
}
} Message: Our TEM was serviced once last year for preventative
} maintenance and a new camera meter. We paid almost $10,000 for our
} service contract. This is a large sum of money for an old scope that
} does not see much wear and tear from too many users. Does anybody
} have a more reasonable service contract for preventative maintenance
} and/or limited service calls for a reliable, but old film TEM? I am
} in the SF Bay area.
}

--
Julian P.S. Smith III
Director, Winthrop Microscopy Facility
Dept. of Biology
Winthrop University
520 Cherry Rd.
Rock Hill, SC 29733

803-323-2111 x6427 (vox)
803-323-3448 (fax)
803-524-2347 (cell)
Research Website www.birdnest.org/smithj
Personal Website www.rociada-east.net


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From: protrain-at-emcourses.com
Date: Mon, 4 Oct 2010 10:09:59 -0500
Subject: [Microscopy] Service contracts

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

Hi

The conversation has been repeated over and over again and, although we run
maintenance courses, there is in my mind nothing better than have a
professional electron microscope maintenance technician look after your
instrument. The greater the number of instruments of the same make that you
maintain the more you learn about them and the easier they become to fix.
Trying to be a casual maintenance technician is tough.

May I add a poignant comment from a retiring service technician in South
Africa -

" If you washing machine breaks down who do you call? If your television
breaks down who do you call? If your computer breaks down who do you call?
If your electron microscope breaks down who do you call, no you fix it
yourself!!??**"

This has always amused me.

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com

-----Original Message-----
X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz]
Sent: 04 October 2010 15:45
To: protrain-at-emcourses.com

Randy, et al,
Another aspect that I see as an independent, and I'm sure the manufacturers
see the same, is that a non-contract instrument often accumulates many small
problems over time. They are ignored, or lived with, because administrators
are loath to spend the money on a "minor" problem or even on "preventive
maintenance".

When the system eventually goes down, all they are willing to pay for is
enough to get the system so it is not "down". Rather a moving target, don't
you think?

Anyway, in these instances the customer is often not satisfied (user, not
administrator) and from a service engineer's standpoint it is VERY
unsatisfying. Our goal is to have things work correctly, reliably and to
specification. A service contract allows us to do what needs to be done
without having to watch the clock.

Sadly, if we do our job well, I often find that the microscopes end up
having not much more than their 2 preventive maintenance calls per year
after a few years because the major issues have been resolved and those,
often quick, preventive visits still can turn up and fix problems that the
user hasn't seen, yet. The bean counters look at that and say, "The
contract is costing much more than those 2 calls per year. Cancel it and
just schedule the preventive maintenance." Of course, the preventive
maintenance calls aren't scheduled because "it's running fine so we don't
need to spend the money." It's enough to make you pull your hair out.
Fortunately, mine still grows back.

DISCLAIMER: Quality Images is a third party (independent) service company
that has been servicing SEMs for over 29 years.

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Monday, October 04, 2010 10:02 AM
To: kenconverse-at-qualityimages.biz


I strongly agree with those who argue in favor of having major instruments
on a manufacturer's service contract. (Note that there are independent
service providers also who, by all accounts, do a very good job----I just
don't have any personal experience with them.) Without a contract, you are
at the mercy of luck and the service providers' hourly rates for service AND
travel, plus expenses. Two or three days or this and you have already spent
as much as a contract would cost. Except for one disastrous detour into the
realm of insurance company maintenance coverage, we've had contracts on our
scopes for forever.

Same goes for ancillary equipment, which often seems to be neglected. In my
experience, it is much harder to get administrators to agree to contracts
for smaller pieces of equipment (coaters, microtomes, etc.), but the running
of a facility is as much dependent upon them as upon the major instruments.
Obviously, if you can't prepare a specimen to get it into a $500,000
microscope because a $50,000 piece of equipment is down, what good is the
scope? Service costs for support equipment can be just as high as for a
major piece.

Case in point: we recently spent about $10,000 to repair a piece of
equipment that had a service contract cost of about $5000. Costs included
$350/hr for travel, which involved an engineer driving from quite a
distance. Travel costs alone were ~$3000. The engineer was here for about
two hours (and did an excellent job, incidentally), then had to order a part
which I then later installed myself.

On the other hand, you can always take the chance and hope that nothing goes
wrong.....

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar:
http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week
&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: PhillipsT-at-missouri.edu
Date: Mon, 4 Oct 2010 10:43:47 -0500
Subject: [Microscopy] Service contracts

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

I am also a big believer in service contracts and appreciate Steve's often insightful comments on the listserver but I will critique his logic here. A service contract isn't like calling someone to fix your washing machine or TV. First, you call after the breakdown - most people avoid pre-paying for service contracts on those devices. Second, a lot of broken TV's sadly need to be treated as disposable items these days since repair guys won't waste their time on smaller sets. It just isn't cost effective. My thesis lab had an old Seimens 101 and no money for a service contract. It was a bitch to keep running but we did it - my brilliant thesis adviser even machined new parts for it one time. But I run an LM core now and would be up a creek without service contracts on my confocals. The bottom line is I disagree with Steve's logic but agree with his conclusion. Tom


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: Monday, October 04, 2010 10:11 AM
To: Phillips, Thomas E.

Hi

The conversation has been repeated over and over again and, although we run
maintenance courses, there is in my mind nothing better than have a
professional electron microscope maintenance technician look after your
instrument. The greater the number of instruments of the same make that you
maintain the more you learn about them and the easier they become to fix.
Trying to be a casual maintenance technician is tough.

May I add a poignant comment from a retiring service technician in South
Africa -

" If you washing machine breaks down who do you call? If your television
breaks down who do you call? If your computer breaks down who do you call?
If your electron microscope breaks down who do you call, no you fix it
yourself!!??**"

This has always amused me.

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com

-----Original Message-----
X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz]
Sent: 04 October 2010 15:45
To: protrain-at-emcourses.com

Randy, et al,
Another aspect that I see as an independent, and I'm sure the manufacturers
see the same, is that a non-contract instrument often accumulates many small
problems over time. They are ignored, or lived with, because administrators
are loath to spend the money on a "minor" problem or even on "preventive
maintenance".

When the system eventually goes down, all they are willing to pay for is
enough to get the system so it is not "down". Rather a moving target, don't
you think?

Anyway, in these instances the customer is often not satisfied (user, not
administrator) and from a service engineer's standpoint it is VERY
unsatisfying. Our goal is to have things work correctly, reliably and to
specification. A service contract allows us to do what needs to be done
without having to watch the clock.

Sadly, if we do our job well, I often find that the microscopes end up
having not much more than their 2 preventive maintenance calls per year
after a few years because the major issues have been resolved and those,
often quick, preventive visits still can turn up and fix problems that the
user hasn't seen, yet. The bean counters look at that and say, "The
contract is costing much more than those 2 calls per year. Cancel it and
just schedule the preventive maintenance." Of course, the preventive
maintenance calls aren't scheduled because "it's running fine so we don't
need to spend the money." It's enough to make you pull your hair out.
Fortunately, mine still grows back.

DISCLAIMER: Quality Images is a third party (independent) service company
that has been servicing SEMs for over 29 years.

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Monday, October 04, 2010 10:02 AM
To: kenconverse-at-qualityimages.biz


I strongly agree with those who argue in favor of having major instruments
on a manufacturer's service contract. (Note that there are independent
service providers also who, by all accounts, do a very good job----I just
don't have any personal experience with them.) Without a contract, you are
at the mercy of luck and the service providers' hourly rates for service AND
travel, plus expenses. Two or three days or this and you have already spent
as much as a contract would cost. Except for one disastrous detour into the
realm of insurance company maintenance coverage, we've had contracts on our
scopes for forever.

Same goes for ancillary equipment, which often seems to be neglected. In my
experience, it is much harder to get administrators to agree to contracts
for smaller pieces of equipment (coaters, microtomes, etc.), but the running
of a facility is as much dependent upon them as upon the major instruments.
Obviously, if you can't prepare a specimen to get it into a $500,000
microscope because a $50,000 piece of equipment is down, what good is the
scope? Service costs for support equipment can be just as high as for a
major piece.

Case in point: we recently spent about $10,000 to repair a piece of
equipment that had a service contract cost of about $5000. Costs included
$350/hr for travel, which involved an engineer driving from quite a
distance. Travel costs alone were ~$3000. The engineer was here for about
two hours (and did an excellent job, incidentally), then had to order a part
which I then later installed myself.

On the other hand, you can always take the chance and hope that nothing goes
wrong.....

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar:
http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week
&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: protrain-at-emcourses.com
Date: Mon, 4 Oct 2010 11:22:41 -0500
Subject: [Microscopy] Service contracts

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

Hi

It was not my quote but I did think it was apt. People do not fix their
household items but expect to fix an instrument that is far more complex
than all these household items put together. Would you agree?

Steve

-----Original Message-----
X-from: Phillips, Thomas E. [mailto:PhillipsT-at-missouri.edu]
Sent: 04 October 2010 16:43
To: protrain-at-emcourses.com; microscopy-at-microscopy.com

Hi

The conversation has been repeated over and over again and, although we run
maintenance courses, there is in my mind nothing better than have a
professional electron microscope maintenance technician look after your
instrument. The greater the number of instruments of the same make that you
maintain the more you learn about them and the easier they become to fix.
Trying to be a casual maintenance technician is tough.

May I add a poignant comment from a retiring service technician in South
Africa -

" If you washing machine breaks down who do you call? If your television
breaks down who do you call? If your computer breaks down who do you call?
If your electron microscope breaks down who do you call, no you fix it
yourself!!??**"

This has always amused me.

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com

-----Original Message-----
X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz]

Sent: 04 October 2010 15:45
To: protrain-at-emcourses.com

Randy, et al,
Another aspect that I see as an independent, and I'm sure the manufacturers
see the same, is that a non-contract instrument often accumulates many small
problems over time. They are ignored, or lived with, because administrators
are loath to spend the money on a "minor" problem or even on "preventive
maintenance".

When the system eventually goes down, all they are willing to pay for is
enough to get the system so it is not "down". Rather a moving target, don't
you think?

Anyway, in these instances the customer is often not satisfied (user, not
administrator) and from a service engineer's standpoint it is VERY
unsatisfying. Our goal is to have things work correctly, reliably and to
specification. A service contract allows us to do what needs to be done
without having to watch the clock.

Sadly, if we do our job well, I often find that the microscopes end up
having not much more than their 2 preventive maintenance calls per year
after a few years because the major issues have been resolved and those,
often quick, preventive visits still can turn up and fix problems that the
user hasn't seen, yet. The bean counters look at that and say, "The
contract is costing much more than those 2 calls per year. Cancel it and
just schedule the preventive maintenance." Of course, the preventive
maintenance calls aren't scheduled because "it's running fine so we don't
need to spend the money." It's enough to make you pull your hair out.
Fortunately, mine still grows back.

DISCLAIMER: Quality Images is a third party (independent) service company
that has been servicing SEMs for over 29 years.

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Monday, October 04, 2010 10:02 AM
To: kenconverse-at-qualityimages.biz


I strongly agree with those who argue in favor of having major instruments
on a manufacturer's service contract. (Note that there are independent
service providers also who, by all accounts, do a very good job----I just
don't have any personal experience with them.) Without a contract, you are
at the mercy of luck and the service providers' hourly rates for service AND
travel, plus expenses. Two or three days or this and you have already spent
as much as a contract would cost. Except for one disastrous detour into the
realm of insurance company maintenance coverage, we've had contracts on our
scopes for forever.

Same goes for ancillary equipment, which often seems to be neglected. In my
experience, it is much harder to get administrators to agree to contracts
for smaller pieces of equipment (coaters, microtomes, etc.), but the running
of a facility is as much dependent upon them as upon the major instruments.
Obviously, if you can't prepare a specimen to get it into a $500,000
microscope because a $50,000 piece of equipment is down, what good is the
scope? Service costs for support equipment can be just as high as for a
major piece.

Case in point: we recently spent about $10,000 to repair a piece of
equipment that had a service contract cost of about $5000. Costs included
$350/hr for travel, which involved an engineer driving from quite a
distance. Travel costs alone were ~$3000. The engineer was here for about
two hours (and did an excellent job, incidentally), then had to order a part
which I then later installed myself.

On the other hand, you can always take the chance and hope that nothing goes
wrong.....

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar:
http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week
&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: PhillipsT-at-missouri.edu
Date: Mon, 4 Oct 2010 12:16:26 -0500
Subject: [Microscopy] Service contracts

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

I am not in full agreement. Lots of scientists, by necessity, are quite mechanically adept. Several years ago, my paraffin tissue processor went out and the company want a 2K+ purchase order before they would come to fix it. Instead, my associate Mike Stanley (now of Chroma) and I took it apart and found a small motor that was broken. The company wanted $1200 for it. Instead, we looked at the motor and called the part manufacturer and got the same thing directly for less than $300. I remember once having my quite expensive Codonics color printer go out. We had no money for a repair so I told my young tech to take it apart with great care. She had never fixed a scientific instrument before this but spent two days disassembling it and photographed every single screw as she removed it so she would be able to figure out how to re-assemble. 2 days later she had repaired it and I don't ever remember her being more proud. Lots of scientists (and others) would think nothing of re-wiring their house, building a deck, or laying a tile floor such as I have done. Clearly fixing an EM is sometimes trickier but I would guess at least 20% of the time the problem is mechanical and could be fixed by a clever individual. But having said all that, I think almost everyone should have their EMs and confocals under service contract. I can think of a dozen times you have given brilliant advice to some listserver poster with an EM problem that enabled them to fix a problem - even with a service contract, one often needs to still do some minor stuff. For example, your advice on HV discharge or maintaining rotary pumps earlier this year. I think your postings are some of the more knowledgeable ones on the listserver and usually select them for the NetNotes column in Microscopy Today. I fully agree with your conclusion on this issue but differ in the underlying logic.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: Steve Chapman [mailto:protrain-at-emcourses.com]
Sent: Monday, October 04, 2010 11:23 AM
To: Phillips, Thomas E.
Cc: Microscopical Soc of America

Hi

The conversation has been repeated over and over again and, although we run
maintenance courses, there is in my mind nothing better than have a
professional electron microscope maintenance technician look after your
instrument. The greater the number of instruments of the same make that you
maintain the more you learn about them and the easier they become to fix.
Trying to be a casual maintenance technician is tough.

May I add a poignant comment from a retiring service technician in South
Africa -

" If you washing machine breaks down who do you call? If your television
breaks down who do you call? If your computer breaks down who do you call?
If your electron microscope breaks down who do you call, no you fix it
yourself!!??**"

This has always amused me.

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com

-----Original Message-----
X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz]

Sent: 04 October 2010 15:45
To: protrain-at-emcourses.com

Randy, et al,
Another aspect that I see as an independent, and I'm sure the manufacturers
see the same, is that a non-contract instrument often accumulates many small
problems over time. They are ignored, or lived with, because administrators
are loath to spend the money on a "minor" problem or even on "preventive
maintenance".

When the system eventually goes down, all they are willing to pay for is
enough to get the system so it is not "down". Rather a moving target, don't
you think?

Anyway, in these instances the customer is often not satisfied (user, not
administrator) and from a service engineer's standpoint it is VERY
unsatisfying. Our goal is to have things work correctly, reliably and to
specification. A service contract allows us to do what needs to be done
without having to watch the clock.

Sadly, if we do our job well, I often find that the microscopes end up
having not much more than their 2 preventive maintenance calls per year
after a few years because the major issues have been resolved and those,
often quick, preventive visits still can turn up and fix problems that the
user hasn't seen, yet. The bean counters look at that and say, "The
contract is costing much more than those 2 calls per year. Cancel it and
just schedule the preventive maintenance." Of course, the preventive
maintenance calls aren't scheduled because "it's running fine so we don't
need to spend the money." It's enough to make you pull your hair out.
Fortunately, mine still grows back.

DISCLAIMER: Quality Images is a third party (independent) service company
that has been servicing SEMs for over 29 years.

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Monday, October 04, 2010 10:02 AM
To: kenconverse-at-qualityimages.biz


I strongly agree with those who argue in favor of having major instruments
on a manufacturer's service contract. (Note that there are independent
service providers also who, by all accounts, do a very good job----I just
don't have any personal experience with them.) Without a contract, you are
at the mercy of luck and the service providers' hourly rates for service AND
travel, plus expenses. Two or three days or this and you have already spent
as much as a contract would cost. Except for one disastrous detour into the
realm of insurance company maintenance coverage, we've had contracts on our
scopes for forever.

Same goes for ancillary equipment, which often seems to be neglected. In my
experience, it is much harder to get administrators to agree to contracts
for smaller pieces of equipment (coaters, microtomes, etc.), but the running
of a facility is as much dependent upon them as upon the major instruments.
Obviously, if you can't prepare a specimen to get it into a $500,000
microscope because a $50,000 piece of equipment is down, what good is the
scope? Service costs for support equipment can be just as high as for a
major piece.

Case in point: we recently spent about $10,000 to repair a piece of
equipment that had a service contract cost of about $5000. Costs included
$350/hr for travel, which involved an engineer driving from quite a
distance. Travel costs alone were ~$3000. The engineer was here for about
two hours (and did an excellent job, incidentally), then had to order a part
which I then later installed myself.

On the other hand, you can always take the chance and hope that nothing goes
wrong.....

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar:
http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week
&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: Drake.Damerau-at-gd-ots.com
Date: Mon, 4 Oct 2010 12:41:43 -0500
Subject: [Microscopy] LaB6 on E-bay - Bad Idea?

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

I new to the listserve, so I thought I would introduce myself. I'm a metallurgist specializing in failure analysis at General Dynamics. Although I've been doing this for 20 years, I just got my first SEM. It's a used AMRAY 1830 with EDS, and I'm very happy to have it.

I'm looking to save my company money. There is a LaB6 filament for my SEM on E-Bay right now. Has anyone bought something like this off of E-Bay? ...or is that a bad idea.

Drake Dämerau
Senior Metallurgist/Lab Manager
Drake.Damerau-at-gd-ots.com
(570) 340-1194
General Dynamics
Ordnance and Tactical Systems




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From: gary-at-gaugler.com
Date: Mon, 4 Oct 2010 12:46:40 -0500
Subject: [Microscopy] Service contracts

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

One factor not discussed so far is the impact of computer
controlled instruments via a PC. This plus surface mounted
ICs makes for a very difficult repair situation in comparison
to simple embedded microprocessors. Coming from long Amray SEM
experience, their earlier systems had socketed ICs and came
with full schematics and components layout. These were
very reliable tools and easy to fix. Later generation tools
are "fixed" via board replacement. This is not a trivial issue
and as such, a contract is important--if not very important.
Some makers do not supply schematics, regardless of whether
they would be of use.

gary g.


At 10:17 AM 10/4/2010, you wrote:



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From: gary-at-gaugler.com
Date: Mon, 4 Oct 2010 12:53:06 -0500
Subject: [Microscopy] Re: LaB6 on E-bay - Bad Idea?

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

It depends on what his reserve is. You also need an 1830
with a gun chamber ion pump. If he is complaining about
getting to E-7 Torr, I wonder why? Will not get there
without the ion pump.

gary g.


At 10:43 AM 10/4/2010, you wrote:



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From: Phil.Ahrenkiel-at-sdsmt.edu
Date: Mon, 4 Oct 2010 12:57:18 -0500
Subject: [Microscopy] Service contracts

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

We could get by with just an annual PM and perhaps an emergency service call every couple of years if the service manuals for the instrument was available. I don't mind repairing our TEM; it's the best way to learn the inner workings. But it is impractical to attempt any serious repair by reverse engineering. My impression is that the service engineers usually follow step-by-step procedures outlined in their service manuals, which include electrical schematics, etc. for more extensive diagnostics. Fortunately, most of the companies seem willing to provide some phone support, but the manual pages are closely protected.

If it would save $10K, I would learn to fix my washing machine, too, but I would still want the service manual. Not much of our research relies on uninterrupted access to a finely tuned washing machine, though.

------------------------------------------
Phil Ahrenkiel, Assistant Professor
Nanoscience and Nanoengineering Ph.D. Program
South Dakota School of Mines and Technology
501 E. Saint Joseph St.
Rapid City, SD 57701 U.S.A.
Office: EP 221
Phone: 605-394-5238, Fax: 605-394-2365
Email: Phil.Ahrenkiel-at-sdsmt.edu

-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: Monday, October 04, 2010 9:15 AM
To: Ahrenkiel, Phil

Hi

The conversation has been repeated over and over again and, although we run
maintenance courses, there is in my mind nothing better than have a
professional electron microscope maintenance technician look after your
instrument. The greater the number of instruments of the same make that you
maintain the more you learn about them and the easier they become to fix.
Trying to be a casual maintenance technician is tough.

May I add a poignant comment from a retiring service technician in South
Africa -

" If you washing machine breaks down who do you call? If your television
breaks down who do you call? If your computer breaks down who do you call?
If your electron microscope breaks down who do you call, no you fix it
yourself!!??**"

This has always amused me.

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com

-----Original Message-----
X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz]
Sent: 04 October 2010 15:45
To: protrain-at-emcourses.com

Randy, et al,
Another aspect that I see as an independent, and I'm sure the manufacturers
see the same, is that a non-contract instrument often accumulates many small
problems over time. They are ignored, or lived with, because administrators
are loath to spend the money on a "minor" problem or even on "preventive
maintenance".

When the system eventually goes down, all they are willing to pay for is
enough to get the system so it is not "down". Rather a moving target, don't
you think?

Anyway, in these instances the customer is often not satisfied (user, not
administrator) and from a service engineer's standpoint it is VERY
unsatisfying. Our goal is to have things work correctly, reliably and to
specification. A service contract allows us to do what needs to be done
without having to watch the clock.

Sadly, if we do our job well, I often find that the microscopes end up
having not much more than their 2 preventive maintenance calls per year
after a few years because the major issues have been resolved and those,
often quick, preventive visits still can turn up and fix problems that the
user hasn't seen, yet. The bean counters look at that and say, "The
contract is costing much more than those 2 calls per year. Cancel it and
just schedule the preventive maintenance." Of course, the preventive
maintenance calls aren't scheduled because "it's running fine so we don't
need to spend the money." It's enough to make you pull your hair out.
Fortunately, mine still grows back.

DISCLAIMER: Quality Images is a third party (independent) service company
that has been servicing SEMs for over 29 years.

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Monday, October 04, 2010 10:02 AM
To: kenconverse-at-qualityimages.biz


I strongly agree with those who argue in favor of having major instruments
on a manufacturer's service contract. (Note that there are independent
service providers also who, by all accounts, do a very good job----I just
don't have any personal experience with them.) Without a contract, you are
at the mercy of luck and the service providers' hourly rates for service AND
travel, plus expenses. Two or three days or this and you have already spent
as much as a contract would cost. Except for one disastrous detour into the
realm of insurance company maintenance coverage, we've had contracts on our
scopes for forever.

Same goes for ancillary equipment, which often seems to be neglected. In my
experience, it is much harder to get administrators to agree to contracts
for smaller pieces of equipment (coaters, microtomes, etc.), but the running
of a facility is as much dependent upon them as upon the major instruments.
Obviously, if you can't prepare a specimen to get it into a $500,000
microscope because a $50,000 piece of equipment is down, what good is the
scope? Service costs for support equipment can be just as high as for a
major piece.

Case in point: we recently spent about $10,000 to repair a piece of
equipment that had a service contract cost of about $5000. Costs included
$350/hr for travel, which involved an engineer driving from quite a
distance. Travel costs alone were ~$3000. The engineer was here for about
two hours (and did an excellent job, incidentally), then had to order a part
which I then later installed myself.

On the other hand, you can always take the chance and hope that nothing goes
wrong.....

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar:
http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week
&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: smalinskas-at-yahoo.com
Date: Mon, 4 Oct 2010 14:27:38 -0500
Subject: [Microscopy] RE: Service contracts

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

Though my experience is not directly with our SEM, we presently have a similar issue with our EDS unit in our corporate climate.

We dropped the service contract on our EDS unit a long time ago for substantial savings. I was proud that I could save the company money, reminding them that we could apply this money to replace the unit when the time came. Well, the time has come, and our EDS unit is in dire need of replacement. As you can probably expect, the company now says that there is no money for replacement. (Where is all that money I saved them?) We are stuck limping along with our old unit.

I strongly recommend maintaining a service contract for those working in a corporate (and perhaps academic) climate. In the past I worked in a small independent lab where I was handy and the lab head was also the one paying for everything. In that case it made sense for us to have no (or minimal) service contract.

Stu Smalinskas, PE
Metallurgist
SKF
Plymouth, Michigan




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From: rosemary.white-at-csiro.au
Date: Mon, 4 Oct 2010 17:34:14 -0500
Subject: [Microscopy] Re: Service contracts

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

Yes, we've had very good value out of our service contract on a confocal
microscope, which includes all parts.

On the other hand, we were quoted $40,000 for a service contract on an SEM,
which seemed excessive to us (more than the confocal contract and did not
include parts, but did include unlimited emergency visits - the contract for
just two service visits a year was less). A service contract from the same
company on light microscopes, while reasonable, does not include travel -
fares and time, or overnight accommodation. Since we are not a hub for any
of these companies (we're in a small town - the capital city of
Australia....) we're always slugged for travel. May not seem much each
time, but we've had trouble with these microscopes, and if you have a few
emergency visits a year it adds up.

So there are good and not so great service contracts....

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au



On 5/10/10 12:06 AM, "TindallR-at-missouri.edu" {TindallR-at-missouri.edu} wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
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}
} I strongly agree with those who argue in favor of having major instruments on
} a manufacturer's service contract. (Note that there are independent service
} providers also who, by all accounts, do a very good job----I just don't have
} any personal experience with them.) Without a contract, you are at the mercy
} of luck and the service providers' hourly rates for service AND travel, plus
} expenses. Two or three days or this and you have already spent as much as a
} contract would cost. Except for one disastrous detour into the realm of
} insurance company maintenance coverage, we've had contracts on our scopes for
} forever.
}
} Same goes for ancillary equipment, which often seems to be neglected. In my
} experience, it is much harder to get administrators to agree to contracts for
} smaller pieces of equipment (coaters, microtomes, etc.), but the running of a
} facility is as much dependent upon them as upon the major instruments.
} Obviously, if you can't prepare a specimen to get it into a $500,000
} microscope because a $50,000 piece of equipment is down, what good is the
} scope? Service costs for support equipment can be just as high as for a major
} piece.
}
} Case in point: we recently spent about $10,000 to repair a piece of equipment
} that had a service contract cost of about $5000. Costs included $350/hr for
} travel, which involved an engineer driving from quite a distance. Travel
} costs alone were ~$3000. The engineer was here for about two hours (and did
} an excellent job, incidentally), then had to order a part which I then later
} installed myself.
}
} On the other hand, you can always take the chance and hope that nothing goes
} wrong.....
}
} Cheers,
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar:
} http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&N
} avType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
}
}
} ==============================Original Headers==============================
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12, 39 -- From prvs=886e9dace=Rosemary.White-at-csiro.au Mon Oct 4 17:34:14 2010
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From: Gregory.Hendricks-at-umassmed.edu
Date: Mon, 4 Oct 2010 19:39:08 -0500
Subject: [Microscopy] viaWWW: Northeast Regional Core Directors Meeting

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Email: Gregory.Hendricks-at-umassmed.edu
Name: Greg Hendricks

Organization: UMass Medical School

Title-Subject: [Filtered] Northeast Regional Core Directors Meeting

Message: Please join us at the fifth annual Northeast Regional Life
Sciences Core Directors (NERLSCD) meeting that will be held October
27-29, 2010, at University of Massachusetts Medical School in
Worcester, Massachusetts.
The NERLSCD 2010 meeting will be a regional forum for core facility
directors and managers to network with colleagues, to learn about
biotechnology advances and applications, and to discuss the
challenges and results of implementing shared research resources.
There will be presentations and discussion forums on operational
issues facing biotechnology core laboratories. There will be
scientific sessions and technical workshops on proteomics, imaging,
and other technologies, including next generation sequencing,
microarrays, metabolomics, optical imaging, flow cytometry, and
bioinformatics. A core facility poster session will offer an
opportunity for learning about regional life sciences shared
resources and services. Participants will be able to post and to
learn of core facility related employment opportunities. Tours will
be given of core facilities. There will be an opening reception and
numerous opportunities to network with colleagues.
We look forward to seeing you at the NERLSCD 2010 meeting!
If you are interested, please contact Susanna Perkins at:
Susanna.Perkins-at-umassmed.edu

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From: bplowman-at-pacific.edu
Date: Mon, 4 Oct 2010 19:39:46 -0500
Subject: [Microscopy] viaWWW: Beam damage to sections

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Email: bplowman-at-pacific.edu
Name: Barbara Plowman

Organization: University of the Pacific, Arthur A. Dugoni School of Dentistry

Title-Subject: [Filtered] Beam damage to sections

Message: Yes, I agree that one can avoid a lot of sections breaking
by spreading the beam as much as possible and going over the grid to
"temper" them before using too much illumination to view them.
It is sort of like what i do when I make tea in a ceramic teapot. I
fill the teapot with hot water and warm it up before putting in the
boiling water. That way it won't break from the shock of the
temperature change.
Raise the amount of illumination gradually and don't zap them all at
once with the beam.

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From: nizets2-at-yahoo.com
Date: Tue, 5 Oct 2010 04:40:32 -0500
Subject: [Microscopy] service contracts

Contents Retrieved from Microscopy Listserver Archives
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I wondered if I would dare go against all this wonderful communitary unity FOR
service contracts from major companies.
Well I still don't know if I'd dare, so let me be careful and make some maths.
For our TEM at least, the service contract would cost about 10% of the price of
the microscope yearly. That means, with the money spared you could buy a new
scope every 10 years (or every 13 years, if you include the first 3 years under
guarantee). This calculation was confirmed by a colleague of mine, which runs a
FEG under service contract.
Now the question is: are 10/13 years so much for microscope? Are they so old
after 10 years, that they need major repairs?
Of course some pieces need replacement: IGPs and the like, but perhaps with a
little bit of training one can do it without the need for a professional. These
pieces have a cost, but it was clear from previous posts that work hours and
travel took a big part in the bill.
I don't even want to think that after 20 years of service contract I could have
2 microscopes instead of a 20 years-old one!! (even under service contract)

Am I over-optimistic? Where am I wrong?

Stephane




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From: Aleksandr.Mironov-at-manchester.ac.uk
Date: Tue, 5 Oct 2010 05:48:19 -0500
Subject: [Microscopy] Sapphire coverslips

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Dear listers,

Do you know any suppliers who can provide me with non standard 50um
sapphire coverslips? I need them to be of 1.9mm in diameter, the
standard ones are 1.4mm, 3mm and 6mm.

Regards,
Alex

--
Dr. Aleksandr Mironov MD, PhD
Senior Experimental Officer
D.1527, M.Smith Building
EM Core Facility, Faculty of Life Sciences
University of Manchester
Oxford Road
Manchester
M13 9PT
UK

Tel. +44-(0)161-275-5645
Fax. +44-(0)161-275-5171
E-mail: Aleksandr.Mironov-at-manchester.ac.uk
http://www.ls.manchester.ac.uk/research/facilities/electronmicroscopy/


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From: Frank_Karl-at-lincolnelectric.com
Date: Tue, 5 Oct 2010 06:45:53 -0500
Subject: [Microscopy] Re: service contracts

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I've read the post and it is clear this will not be resolved but I do have
a few things I'd like to rant about..

I have often seen the post that seem gloat over the undergrad who spent two
days taking apart and solving a problem with a scientific instrument. It
is something to be proud of, but you let them fiddle with it because what
was the risk, as they say, it was already broken.

How happy would you be if you were paying that person $50.00 per hour (not
unreasonable including benefits) to spend 16 hours in which billable
projects were piling up and still not be sure they would fix it? And what
if in the process, a resister cracks internally from twisting a circuit
board out, or a critical screw stripped, or a leaded glass port slips and
chips an edge. Humm, that simple repair job may mean the instrument is
down for a week. What do think will happen to your salery-bonus-raise when
the CEO submits problem from a customer they are interested in? Especially
the customer whose account helped make him CEO.

Go service contract, pay the money, get the knowledge and the protective
edge they provide. Scope down for a week? The service guy is doing
everything he can to breath life back into it.

I agree, I don't repair my wash machine and I don't repair my own tires.
And I think these are simple fixes compared to my SEM. So why would try to
fix my SEM or TEM when the y-scan goes out or the stigmator dies?

Rant mode off......

stay safe..........
Frank


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From: john.robson-at-boehringer-ingelheim.com
Date: Tue, 5 Oct 2010 07:23:51 -0500
Subject: [Microscopy] RE: service contracts

Contents Retrieved from Microscopy Listserver Archives
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Depending on the instrument design and the types of samples being analyzed
maintenance, even the routine (to be expected) stuff, can become quite time
consuming which comes at a price even if you maintain the equipment yourself.
Not to mention the infrequent system failures that crop up throughout the
useful life of the instrument. I get paid to deliver result, that's the
bottom line, the more time I spend learning how to inefficiently repair
equipment the less efficient I am at meeting performance expectations. Now I
can go hourly for routine stuff which would save us 20-30% off the contract
rate but any failures would eat into those savings. I would also risk losing
priority status with the manufacture which would further cut into up-time and
efficiency. To contract or not to contract.......that is the question? And
the answer is....................it depends on your situation but in our case
given our equipment, samples, staffing, and priorities a service contract is
the answer.

PS I have a 20 year old FEG that works better than the day it was installed.
If you do the math it was much cheaper to keep it under contract.

Regards John


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4, 31 -- To: {Microscopy-at-microscopy.com}
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From: bozzola-at-siu.edu
Date: Tue, 5 Oct 2010 07:57:03 -0500
Subject: [Microscopy] Re: service contracts

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

These figures sound reasonable to me, Stephane. Based on an informal
poll I made some time ago, it appears the life expectancy of a new EM
is about 15 years (though some would say 10-12 yrs). I was shocked by
this figure, since I have worked with some perfectly fine instruments
nearly twice that age. The older generation of instruments were
simpler, with mechanical adjustments, relatively simple circuitry and
no computer control. With the introduction of computerized controls
(for convenience of operation), and a booming economy, there was a
paradigm shift to what we have presently.

With the economic challenges we face, I believe OEMs should consider
reintroducing a generation of simpler (reliable, less expensive)
instruments. Most biologists, especially at smaller colleges or
research institutions, really do not need the burden of keeping high
end instruments running, especially in a teaching situation. Most EM
work is not being done at the highest resolutions, certainly in the
biological area, so the high end instruments are really over-kill.

There is an obvious need for high end instruments in some situations,
but not everyone needs (or can afford to drive) a BMW when a Camry
would fulfill most of their requirements.

Does anyone remember the Philips 201? Imagine that instrument with a
reasonably priced, digital camera! It can be done, for sure, and my
hope is that some enterprising company would make this step.

I will now duck and cover .......


--
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730

On Tue, Oct 5, 2010 at 4:41 AM, {nizets2-at-yahoo.com} wrote:
}
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} I wondered if I would dare go against all this wonderful communitary unity FOR
} service contracts from major companies.
} Well I still don't know if I'd dare, so let me be careful and make some maths.
} For our TEM at least, the service contract would cost about 10% of the price of
} the microscope yearly. That means, with the money spared you could buy a new
} scope every 10 years (or every 13 years, if you include the first 3 years under
} guarantee). This calculation was confirmed by a colleague of mine, which runs a
} FEG under service contract.
} Now the question is: are 10/13 years so much for microscope? Are they so old
} after 10 years, that they need major repairs?
} Of course some pieces need replacement: IGPs and the like, but perhaps with a
} little bit of training one can do it without the need for a professional. These
} pieces have a cost, but it was clear from previous posts that work hours and
} travel took a big part in the bill.
} I don't even want to think that after 20 years of service contract I could have
} 2 microscopes instead of a 20 years-old one!! (even under service contract)
}
} Am I over-optimistic? Where am I wrong?
}
} Stephane
}
}


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From: oshel1pe-at-cmich.edu
Date: Tue, 5 Oct 2010 08:15:48 -0500
Subject: [Microscopy] Re: service contracts

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

Stephane,

I'd say you were over-optimistic. What administrator is going to
allow you to accumulate that "10% savings every year" for 10 years?
And then spend it on a new instrument, instead of writing a grant for
the money?
Unless you own your own company, meaning you can make such a
decision, any money you save by not buying a service contract is
*very* likely to be spent elsewhere by whoever does own the company
or runs the university/institute, etc..

Plus, I'd also suggest shopping more. I've been recently pricing TEMs
(grant optimism), and service contracts from most companies are 8-12%
of the purchase price, but not all. One EM manufacturer in particular
is much lower. Less than 1/3 of the other companies (by actual $
quote for the same coverage). And from experience, I'd rate their
service as the best. A major reason for this is business models: most
EM companies service departments are for-profit companies or
divisions, with profit targets. The service department of the lower
cost company is not, hence the lower price.
Caveat: since I'm Tech Ed for Microscopy Today, I don't want to
mention companies. But it's easy enough to check this - get quotes
from sales reps. Or listen to them waffle when you try to get a
quote, which is even more fun.
And ... this applies in the US, possibly also Canada. Europe, I don't know.

Phil

} I wondered if I would dare go against all this wonderful communitary unity FOR
} service contracts from major companies.
} Well I still don't know if I'd dare, so let me be careful and make some maths.
} For our TEM at least, the service contract would cost about 10% of
} the price of
} the microscope yearly. That means, with the money spared you could buy a new
} scope every 10 years (or every 13 years, if you include the first 3
} years under
} guarantee). This calculation was confirmed by a colleague of mine,
} which runs a
} FEG under service contract.
} Now the question is: are 10/13 years so much for microscope? Are they so old
} after 10 years, that they need major repairs?
} Of course some pieces need replacement: IGPs and the like, but perhaps with a
} little bit of training one can do it without the need for a
} professional. These
} pieces have a cost, but it was clear from previous posts that work hours and
} travel took a big part in the bill.
} I don't even want to think that after 20 years of service contract I
} could have
} 2 microscopes instead of a 20 years-old one!! (even under service contract)
}
} Am I over-optimistic? Where am I wrong?
}
} Stephane

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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From: benada-at-biomed.cas.cz
Date: Tue, 5 Oct 2010 08:41:54 -0500
Subject: [Microscopy] Re: viaWWW: high beam sensitive sample

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

Dear Pim,
You can try to coat your section on the grid with very thin layer of carbon. It might help you.
A long time ago we have use it for freeze-dried cryosections and it was working well.

Best regards Oldrich


On Saturday 02 of October 2010 15:00:37 gnonano-at-yahoo.com wrote:
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} Name: Pim
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} Organization: PhD
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} Title-Subject: [Filtered] high beam sensitive sample
}
} Message: My sample is polymer blend (PS/EVOH) and EVOH phase is high
} beam sensitive. What should I do to get rid of this problem?
}
} This is my sample preparation ; Sample was microtomed to 50-70 nm
} thickness and put in a copper grid. TEM with 120 KV was used to
} examined.
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From: smithj-at-winthrop.edu
Date: Tue, 5 Oct 2010 09:00:03 -0500
Subject: [Microscopy] service contracts

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

I'm right with John on this one, especially if the 201 has the
optional side-entry gonio stage.

Computer-control is nice, but I really wish the manufacturers would
design instruments that can be run from a web browser. An ethernet plug
is pretty compatible much compatible with anything.

Julian


On 10/5/10 9:03 AM, bozzola-at-siu.edu wrote:
}
}
} ----------------------------------------------------------------------------
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} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} These figures sound reasonable to me, Stephane. Based on an informal
} poll I made some time ago, it appears the life expectancy of a new EM
} is about 15 years (though some would say 10-12 yrs). I was shocked by
} this figure, since I have worked with some perfectly fine instruments
} nearly twice that age. The older generation of instruments were
} simpler, with mechanical adjustments, relatively simple circuitry and
} no computer control. With the introduction of computerized controls
} (for convenience of operation), and a booming economy, there was a
} paradigm shift to what we have presently.
}
} With the economic challenges we face, I believe OEMs should consider
} reintroducing a generation of simpler (reliable, less expensive)
} instruments. Most biologists, especially at smaller colleges or
} research institutions, really do not need the burden of keeping high
} end instruments running, especially in a teaching situation. Most EM
} work is not being done at the highest resolutions, certainly in the
} biological area, so the high end instruments are really over-kill.
}
} There is an obvious need for high end instruments in some situations,
} but not everyone needs (or can afford to drive) a BMW when a Camry
} would fulfill most of their requirements.
}
} Does anyone remember the Philips 201? Imagine that instrument with a
} reasonably priced, digital camera! It can be done, for sure, and my
} hope is that some enterprising company would make this step.
}
} I will now duck and cover .......
}
}


--
Julian P.S. Smith III
Director, Winthrop Microscopy Facility
Dept. of Biology
Winthrop University
520 Cherry Rd.
Rock Hill, SC 29733

803-323-2111 x6427 (vox)
803-323-3448 (fax)
803-524-2347 (cell)
Research Website www.birdnest.org/smithj
Personal Website www.rociada-east.net


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From: kraftpiano-at-gmail.com
Date: Tue, 5 Oct 2010 09:16:16 -0500
Subject: [Microscopy] Re: service contracts

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

I agree with Julian about having something easy to control from any machine, however based on my extensive IT experience, I can assure you that once you plug an ethernet cable into something you leave it wide open for all sorts of problems.

Imagine if you had your remotely operated TEM (Without a local console) compromised by a hacker who decided to "Exercise" the beam current and start blowing filaments...

Personally, I've always been a fan of computer control of things with an isolated machine, but allowing for manual takeover if needed. I don't think an instrument should be permanently tied to a specific computer. I've seen way too much instability in both individual computers and computer technology as a whole.

Just my two cents...

--Justin A. Kraft

On Oct 5, 2010, at 10:04 AM, smithj-at-winthrop.edu wrote:

}
}
}
} ----------------------------------------------------------------------------
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} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} I'm right with John on this one, especially if the 201 has the
} optional side-entry gonio stage.
}
} Computer-control is nice, but I really wish the manufacturers would
} design instruments that can be run from a web browser. An ethernet plug
} is pretty compatible much compatible with anything.
}
} Julian
}
}
} On 10/5/10 9:03 AM, bozzola-at-siu.edu wrote:
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } These figures sound reasonable to me, Stephane. Based on an informal
} } poll I made some time ago, it appears the life expectancy of a new EM
} } is about 15 years (though some would say 10-12 yrs). I was shocked by
} } this figure, since I have worked with some perfectly fine instruments
} } nearly twice that age. The older generation of instruments were
} } simpler, with mechanical adjustments, relatively simple circuitry and
} } no computer control. With the introduction of computerized controls
} } (for convenience of operation), and a booming economy, there was a
} } paradigm shift to what we have presently.
} }
} } With the economic challenges we face, I believe OEMs should consider
} } reintroducing a generation of simpler (reliable, less expensive)
} } instruments. Most biologists, especially at smaller colleges or
} } research institutions, really do not need the burden of keeping high
} } end instruments running, especially in a teaching situation. Most EM
} } work is not being done at the highest resolutions, certainly in the
} } biological area, so the high end instruments are really over-kill.
} }
} } There is an obvious need for high end instruments in some situations,
} } but not everyone needs (or can afford to drive) a BMW when a Camry
} } would fulfill most of their requirements.
} }
} } Does anyone remember the Philips 201? Imagine that instrument with a
} } reasonably priced, digital camera! It can be done, for sure, and my
} } hope is that some enterprising company would make this step.
} }
} } I will now duck and cover .......
} }
} }
}
}
} --
} Julian P.S. Smith III
} Director, Winthrop Microscopy Facility
} Dept. of Biology
} Winthrop University
} 520 Cherry Rd.
} Rock Hill, SC 29733
}
} 803-323-2111 x6427 (vox)
} 803-323-3448 (fax)
} 803-524-2347 (cell)
} Research Website www.birdnest.org/smithj
} Personal Website www.rociada-east.net
}
}
} ==============================Original Headers==============================
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From: Torsten.Fregin-at-zoologie.uni-hamburg.de
Date: Tue, 5 Oct 2010 09:29:16 -0500
Subject: [Microscopy] Re: service contracts

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


} Personally, I've always been a fan of computer control of things with an isolated machine,
but allowing for manual takeover if needed. I don't think an instrument should be
permanently tied to a specific computer.

That reminds me that having a service contract might mean to get the new computer with
WinXP for nothing and not having the contract not getting it or to hear "there is no such thing
like a computer upgrade because the frame grabber does not work with another computer -
we can serve the confocal but..." (and get stuck with Win NT and the frame grabber even
works with 64 bit Windows)... And then you end up buying a new "cheap" confocal which is
not much faster (533 Hz vs. 400 Hz ?)...

Torsten





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From: nicholls-at-uic.edu
Date: Tue, 5 Oct 2010 09:31:21 -0500
Subject: [Microscopy] Midwest Association of Core Directors

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

The first meeting of Midwest Association of Core Directors will be held
Thursday-Saturday October 21-23, 2010 at the Crowne Plaza Hotel in Chicago.
Several Vendors will provide user/interest meetings on Thursday (see web
site for details). The MWACD sessions will begin with a reception 6:00 PM
Thursday evening at Jak's Tap, 901 West Jackson Boulevard. Sessions will
run all day Friday and conclude 2:00 Saturday. Friday sessions will focus
on general core lab concerns such as LIMS systems, informatics, funding,
NIH/NCRR, ending the day with a core facility poster session. Saturday will
feature breakout sessions for various technology interest groups, and a
final meeting to provide an open forum on future meetings.

More information and schedule at
http://mwacd.abrf.org/index.cfm/events.details?eventID=130 .

Alan W Nicholls, PhD
Interim Associate Director - RRC
Director of Research Service Facility (Electron Microscopy)
Research Resources Center - East (M/C 337)
Room 110 Science and Engineering South Building
The University of Illinois at Chicago
845 West Taylor St
Chicago, IL 60607-7058

Tel: 312 996 1227
Fax: 312 996 8091
Office: Room 110

Web site www.rrc.uic.edu


==============================Original Headers==============================
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From: kenconverse-at-qualityimages.biz
Date: Tue, 5 Oct 2010 12:26:36 -0500
Subject: [Microscopy] Service contracts

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

Hi Peggy,
I was looking at yours (and very much like your points) and it didn't seem
to be directly talking to me, but only sent to me. Here's what I do.

Hit Reply and that will put up the address for whoever sent the message,
then add MSA Listserver (microscopy-at-microscopy.com) . That's the road to
fame, if not fortune.

Franklin is a pretty good hike from Boston and Campobello a hike beyond
that. My brother and his family went up there a number of summers kayaking
with the seals and fin whales. He says that's rush!

I bet the foliage was pretty good. We haven't hit prime in the southern
part of the state, yet, but it's getting close.

If you should ever stray into the Lakes Region, drop by.

Ken

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: Sherwood, Margaret [mailto:MSHERWOOD-at-PARTNERS.ORG]
Sent: Tuesday, October 05, 2010 11:56 AM
To: kenconverse-at-qualityimages.biz

Randy, et al,
Another aspect that I see as an independent, and I'm sure the manufacturers
see the same, is that a non-contract instrument often accumulates many small
problems over time. They are ignored, or lived with, because administrators
are loath to spend the money on a "minor" problem or even on "preventive
maintenance".

When the system eventually goes down, all they are willing to pay for is
enough to get the system so it is not "down". Rather a moving target, don't
you think?

Anyway, in these instances the customer is often not satisfied (user, not
administrator) and from a service engineer's standpoint it is VERY
unsatisfying. Our goal is to have things work correctly, reliably and to
specification. A service contract allows us to do what needs to be done
without having to watch the clock.

Sadly, if we do our job well, I often find that the microscopes end up
having not much more than their 2 preventive maintenance calls per year
after a few years because the major issues have been resolved and those,
often quick, preventive visits still can turn up and fix problems that the
user hasn't seen, yet. The bean counters look at that and say, "The
contract is costing much more than those 2 calls per year. Cancel it and
just schedule the preventive maintenance." Of course, the preventive
maintenance calls aren't scheduled because "it's running fine so we don't
need to spend the money." It's enough to make you pull your hair out.
Fortunately, mine still grows back.

DISCLAIMER: Quality Images is a third party (independent) service company
that has been servicing SEMs for over 29 years.

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Monday, October 04, 2010 10:02 AM
To: kenconverse-at-qualityimages.biz


I strongly agree with those who argue in favor of having major instruments
on a manufacturer's service contract. (Note that there are independent
service providers also who, by all accounts, do a very good job----I just
don't have any personal experience with them.) Without a contract, you are
at the mercy of luck and the service providers' hourly rates for service AND
travel, plus expenses. Two or three days or this and you have already spent
as much as a contract would cost. Except for one disastrous detour into the
realm of insurance company maintenance coverage, we've had contracts on our
scopes for forever.

Same goes for ancillary equipment, which often seems to be neglected. In my
experience, it is much harder to get administrators to agree to contracts
for smaller pieces of equipment (coaters, microtomes, etc.), but the running
of a facility is as much dependent upon them as upon the major instruments.
Obviously, if you can't prepare a specimen to get it into a $500,000
microscope because a $50,000 piece of equipment is down, what good is the
scope? Service costs for support equipment can be just as high as for a
major piece.

Case in point: we recently spent about $10,000 to repair a piece of
equipment that had a service contract cost of about $5000. Costs included
$350/hr for travel, which involved an engineer driving from quite a
distance. Travel costs alone were ~$3000. The engineer was here for about
two hours (and did an excellent job, incidentally), then had to order a part
which I then later installed myself.

On the other hand, you can always take the chance and hope that nothing goes
wrong.....

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar:
http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week
&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: TindallR-at-missouri.edu
Date: Tue, 5 Oct 2010 14:01:13 -0500
Subject: [Microscopy] Chloroform---Thanks!

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Thanks to all who responded to my plea to set my mind at ease over chloroform stretching of sections. I'm pretty sure at this point that I just over-gassed the poor little things.

And thanks for the concern over my liver, Tom. Now where was that when I was imbibing ethanol variants at your house? Hmmmmm?

Cheers all,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
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From: michael.cammer-at-med.nyu.edu
Date: Tue, 5 Oct 2010 20:04:06 -0500
Subject: [Microscopy] viaWWW: converter to put Olympus objective on Nikon body

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Email: michael.cammer-at-med.nyu.edu
Name: Michael Cammer

Organization: NYU Skirball

Title-Subject: [Filtered] converter to put Olympus objective on Nikon body

Message: Does anyone know where we may find a glassless converter
(just a metal ring with threads) to put an Olympus objective on a
Nikon Eclipse microscope?

Thank you very much.

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From: acalhoun-at-bidmc.harvard.edu
Date: Tue, 5 Oct 2010 20:04:35 -0500
Subject: [Microscopy] viaWWW: Reusing Grids

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Email: acalhoun-at-bidmc.harvard.edu
Name: Andrea Calhoun

Organization: Beth Israel Deaconess Medical Center

Title-Subject: [Filtered] Reusing Grids

Message: You know how we strive for perfection, right? Well, in my
imperfect times, my hand coated formvar grids are sometimes imperfect
as well (popped coating in few or some grid spaces). This leaves me
with quite a few grids that I "can't use".

Does anyone clean and re-coat their grids? I read somewhere that
chloroform is one way to remove formvar. Does anyone else do this?
Are there other solvents that I could use? Is this a bad idea?

Any input would be much appreciated (especially by my supply budget).

Thanks!

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From: vakimler-at-med.wayne.edu
Date: Tue, 5 Oct 2010 20:04:59 -0500
Subject: [Microscopy] viaWWW: Double Label Immunogold/LR White Hard and Medium

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Double Label Immunogold/LR White Hard and Medium

Message: 1.) When looking for two antigens with immunogold, should
the larger probe be targeted against the more abundant antigen, or
the larger (greater size/molecular weight) antigen in the cell?

Also, some antigens are stationary (membrane bound) and others
translocate. Does the location and "activity" of the antigen
contribute to determining which size of probe to use?

2.) Is there a difference between LR White Hard and LR White Medium,
with respect to antigenicity and tissue/cell shrinkage? Or are they
the same in these regards?

Thanks,
Vickie Kimler





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From: msherwood-at-partners.org
Date: Tue, 5 Oct 2010 20:05:26 -0500
Subject: [Microscopy] viaWWW: Fixation and Processing of Adipose Tissue for TEM

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Email: msherwood-at-partners.org
Name: Margaret E. Sherwood

Organization: Massachusetts General Hospital-Wellman Center

Title-Subject: [Filtered] Re: Fixation and Processing of Adipose
Tissue for TEM

Message: To all:

I have never fixed or processed adipose tissue. Would someone send me
a protocol to do this? (I found an old reference that indicated
fixing in a mix of glutaraldehyde and osmium in buffer).

The investigator wants to look for mitochondria on the edges of the adipose.

Thanks!
Peggy

Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDR 214)
Massachusetts General Hospital
55 Fruit Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood-at-partners.org=20



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From: bozzola-at-siu.edu
Date: Tue, 5 Oct 2010 22:04:08 -0500
Subject: [Microscopy] Re: viaWWW: Reusing Grids

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

Yes, you could clean and re-use the grids. However, bent or warped
grids should be avoided since a new Formvar film would not adhere very
well. Or, you could use the uncoated grids to collect sections
directly.

-
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


On Tue, Oct 5, 2010 at 8:05 PM, {acalhoun-at-bidmc.harvard.edu} wrote:
}
}

}
} Email: acalhoun-at-bidmc.harvard.edu
} Name: Andrea Calhoun
}
} Organization: Beth Israel Deaconess Medical Center
}
} Title-Subject: [Filtered] Reusing Grids
}
} Message: You know how we strive for perfection, right? Well, in my
} imperfect times, my hand coated formvar grids are sometimes imperfect
} as well (popped coating in few or some grid spaces). This leaves me
} with quite a few grids that I "can't use".
}
} Does anyone clean and re-coat their grids? I read somewhere that
} chloroform is one way to remove formvar. Does anyone else do this?
} Are there other solvents that I could use? Is this a bad idea?
}
} Any input would be much appreciated (especially by my supply budget).
}
} Thanks!
}



-


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From: tomas.hrncir-at-tescan.cz
Date: Wed, 6 Oct 2010 01:31:37 -0500
Subject: [Microscopy] Re: Sapphire coverslips

Contents Retrieved from Microscopy Listserver Archives
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} Do you know any suppliers who can provide me with non standard 50um
} sapphire coverslips? I need them to be of 1.9mm in diameter, the
} standard ones are 1.4mm, 3mm and 6mm.

Alex,
try to ask Crytur for custom made sapphire products:

http://www.crytur.cz

Best regards,
Tomas

--
Tomas Hrncir, Ph.D.
R&D - Physics
Tescan
Libusina trida 21
623 00 Brno - CZ
Phone: +420 547 130 468
Fax: +420 547 130 415
http://www.tescan.cz


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From: PhillipsT-at-missouri.edu
Date: Wed, 6 Oct 2010 07:02:19 -0500
Subject: [Microscopy] viaWWW: Double Label Immunogold/LR White Hard and Medium

Contents Retrieved from Microscopy Listserver Archives
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There are numerous papers showing the larger gold size you use, the lower the labeling efficiency. Therefore always label your more abundant antigen with the larger gold. There is probably no difference in membrane bound vs cytoplasmic antigens per se. In a comparison of any two targets, the antigenicity of one is often better retained but I don't know if you could predict it based on membrane-bound vs. cytoplasmic. I have never compared LRW hard vs medium for relative antigenicity but have often compared LR Gold vs LR White vs Lowicryl K4M vs Lowicryl HM20. It is antigen dependent but I usually go LRG } HM20 } K4M. HM20 is much easier to section with little difference in immunoreactivity compared to K4M. LRW was designed for osmicated tissues so isn't generally included in a comparison with the others which are UV polymerized and not for osmicated tissues. Osmication will significantly decrease your success rate. Good luck.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Double Label Immunogold/LR White Hard and Medium

Message: 1.) When looking for two antigens with immunogold, should
the larger probe be targeted against the more abundant antigen, or
the larger (greater size/molecular weight) antigen in the cell?

Also, some antigens are stationary (membrane bound) and others
translocate. Does the location and "activity" of the antigen
contribute to determining which size of probe to use?

2.) Is there a difference between LR White Hard and LR White Medium,
with respect to antigenicity and tissue/cell shrinkage? Or are they
the same in these regards?

Thanks,
Vickie Kimler





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22, 35 -- From PhillipsT-at-missouri.edu Wed Oct 6 07:02:19 2010
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From: W.Muss-at-salk.at
Date: Wed, 6 Oct 2010 07:19:14 -0500
Subject: [Microscopy] Re: Sapphire coverslips [special type]

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Good afternoon,

dear Alexandr,
just to point you to the following company website (DISCLAIMER: no affiliation, no financial interest, just providing collegial information):

==} http://www.escoproducts.com/products/sapphire-optical-circle.php?show=2

==} Sapphire coverslips, Product number: G107020 50µm thickness, 19.05 mm diameter, in stock

Hoping that this is what you were looking for,

Best wishes and regards

Wolfgang MUSS PhD
Salzburg, Austria



} -----Ursprüngliche Nachricht-----
} Von: Aleksandr.Mironov-at-manchester.ac.uk
} [mailto:Aleksandr.Mironov-at-manchester.ac.uk]
} Gesendet: Dienstag, 05. Oktober 2010 12:51
} An: Muß Wolfgang
} Betreff: [Microscopy] Sapphire coverslips
}
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} Dear listers,
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} Do you know any suppliers who can provide me with non standard 50µm sapphire coverslips?
}
} I need them to be of 1.9mm in diameter, the standard ones are 1.4mm, 3mm and 6mm.
}
} Regards,
} Alex
}
} --
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From: srannecherie-at-sydney.edu.au
Date: Wed, 6 Oct 2010 08:04:54 -0500
Subject: [Microscopy] viaWWW: Job Advertisement - ATOM PROBE ENGINEER

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Name: Sydney Recruitment

Organization: The University of Sydney

Title-Subject: [Filtered] Job Advertisement - ATOM PROBE ENGINEER

Message: ATOM PROBE ENGINEER
DVC RESEARCH - Australian Centre for Microscopy & Microanalysis-ACMM
REFERENCE NO. 2616 /0810

ï Be part of the largest microscopy centre of its type in Australia
ï Opportunity to work on newly emerging
atomic resolution imaging technique
ï Remuneration package- $67,962.37- $76,152.79
Closing date Sunday 24th October 2010

The University of Sydney is Australia's premier
University with an outstanding global reputation
for academic and research excellence, and employs
over 6,800 permanent staff supporting over 46,000
students.

The Australian Centre for Microscopy and
Microanalysis (ACMM) is a world-leading research
facility at the university, with excellent staff
that provide training and support for a large and
diverse user community from across Sydney and
beyond. The ACMM (sydney.edu.au/acmm) also
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international characterisation communities and
serves as national headquarters of the Australian
Microscopy & Microanalysis Research Facility
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The atom probe is emerging as one of the most
exciting areas of all areas of imaging and
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From: TindallR-at-missouri.edu
Date: Wed, 6 Oct 2010 08:54:45 -0500
Subject: [Microscopy] viaWWW: Reusing Grids

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Ethylene dichloride (dichloroethane) works to clean grids. This is generally what formvar is dissolved in when you buy it in solution.

Cheers,
Randy

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Email: acalhoun-at-bidmc.harvard.edu
Name: Andrea Calhoun

Organization: Beth Israel Deaconess Medical Center

Title-Subject: [Filtered] Reusing Grids

Message: You know how we strive for perfection, right? Well, in my
imperfect times, my hand coated formvar grids are sometimes imperfect
as well (popped coating in few or some grid spaces). This leaves me
with quite a few grids that I "can't use".

Does anyone clean and re-coat their grids? I read somewhere that
chloroform is one way to remove formvar. Does anyone else do this?
Are there other solvents that I could use? Is this a bad idea?

Any input would be much appreciated (especially by my supply budget).

Thanks!

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From: Leigh.Estrada-at-fei.com
Date: Wed, 6 Oct 2010 10:52:34 -0500
Subject: [Microscopy] FEI - Applications Engineer Position - SEM/FIB/SDB

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

FEI is searching for an Applications Engineer to join the Scanning Electron Microscope / Small DualBeam(tm) (SEM/SDB) applications team.  This customer focused position is ideal for an experienced technologist wanting to be involved with a dynamic team and exposed to a constant variety of customer application areas.  PHD level of education is preferred with direct experience and knowledge of FIB/SEM or SDB applications including TEM sample prep, cross section analysis, and ultra high resolution imaging.  Knowledge of advanced technology diagnostics as well as fault finding techniques, operational optimization, experience of analytical equipment; specifically electron/ion microscopes and various energy dispersive spectrometers is required.  The role will be located at FEI headquarters in Hillsboro, Oregon.   Ability to travel 50% is required (both domestic and international). 

The full position description can be reviewed here: http://careers.fei.com/DetailFEI.asp?jobid=fei2391

Please apply online using the following link:  http://careers.fei.com/applyFEI2.asp?fei?fei2391?lestrada?9#


Leigh Estrada
Talent Acquisition
FEI Company
leigh.estrada-at-fei.com
503-726-7574 - direct
www.fei.com



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From: Leigh.Estrada-at-fei.com
Date: Wed, 6 Oct 2010 10:54:27 -0500
Subject: [Microscopy] FEI - Product Marketing Engineer Position

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

FEI is searching for a senior level Product Marketing Engineer.  This role is responsible for developing business plans, marketing strategy, and forecasts for assigned product lines, maintaining a strong understanding of customer technical requirements for existing and future products. This position identifies, evaluates, and recommends marketing opportunities consistent with product line objectives.  Ability to develop sales support materials including brochures, applications notes, product data sheets, competitive matrices, and technical presentations.   Master's or PHD degree preferred.  Direct experience with SEMs or FIBs is required.  This position is ideal for an experienced professional wanting to be involved with a dynamic team and exposed to a constant variety of customer application areas.   This role will be located in Hillsboro, Oregon.  Ability to travel 50% (domestically and internationally) is required.

Full job description may be viewed here:  http://careers.fei.com/DetailFEI.asp?jobid=fei2319

Please apply online using the following link:  http://careers.fei.com/applyFEI2.asp?fei?fei2319?lestrada?9

Leigh Estrada
Talent Acquisition
FEI Company
leigh.estrada-at-fei.com
503-726-7574 - direct
www.fei.com



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From: maryflet-at-interchange.ubc.ca
Date: Wed, 6 Oct 2010 11:04:48 -0500
Subject: [Microscopy] viaWWW: Reusing Grids

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

Dear Andrea,
I make my own carbon-coated grids, using collodion to make the initial film
and dissolving it later in chloroform. If I botch a batch, I clean the grids
of collodion in acetone. Just throw them in a beaker with acetone and
ultrasound for a few seconds.
Regards,

Mary Fletcher
Electron Microscopist
Materials Eng. UBC
#309 - 6350 Stores Road
Vancouver, B.C. V6T 1Z4
Canada
Tel: 604-822-5648
Fax: 604-822-3619
email: maryflet-at-interchange.ubc.ca


-----Original Message-----
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Sent: October 5, 2010 6:21 PM
To: maryflet-at-interchange.ubc.ca

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Email: acalhoun-at-bidmc.harvard.edu
Name: Andrea Calhoun

Organization: Beth Israel Deaconess Medical Center

Title-Subject: [Filtered] Reusing Grids

Message: You know how we strive for perfection, right? Well, in my
imperfect times, my hand coated formvar grids are sometimes imperfect
as well (popped coating in few or some grid spaces). This leaves me
with quite a few grids that I "can't use".

Does anyone clean and re-coat their grids? I read somewhere that
chloroform is one way to remove formvar. Does anyone else do this?
Are there other solvents that I could use? Is this a bad idea?

Any input would be much appreciated (especially by my supply budget).

Thanks!

Login Host: 134.174.110.7
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From: rintugr-at-gmail.com
Date: Wed, 6 Oct 2010 14:23:20 -0500
Subject: [Microscopy] Wood microscopy

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

Dear Colleagues,

A post doctoral fellow in my department has some wood samples for
imaging (surface only, no sectioning). I was thinking of doing SEM of
a highly polished sample to see the individual rings and the cell
structure. However, it is always best to ask the experts for the right
approach. They have a large number of samples and any help in this
matter will be very much appreciated.

Given below is her email message and the dropbox link has the image of
the wooden planchets.

Thank you,

Soumitra

************************************************************
Soumitra Ghoshroy
Director, Electron Microscopy Center
Research Associate Professor, Biology
University of South Carolina
Columbia, SC 29208
803-777-7085 (office)
http://www.emc.sc.edu
************************************************************

Dear Soumitra,
my colleagues and I would like to ask about any techniques to make
birch year rings visible.
We have wood samples taken from trees growing in different ecological
conditions and we need to measure year rings. We did this with pine
samples, but it is really difficult to find year rings in birch wood
samples. We have a guess, that in is possible to color them in
someway. The samples are putted into wooden planchettes and fixed
there with glue. We need these samples for future mass-spectrometry
analysis, so the task is to make year rings visible but not destroy
samples.

Sample lengths is about 20-30 centimeters, they are air-dry, photo is added.

http://dl.dropbox.com/u/7737489/Wood_Cores.pdf

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From: mike.bode-at-resaltatech.com
Date: Wed, 6 Oct 2010 15:31:45 -0500
Subject: [Microscopy] Wood microscopy

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

I prepared one reply then carefully read the original post, so now my response is slightly different.

I suppose embedding and polishing is off limits if they want to do mass spec on the same samples later.

It could be that the rings might be visible within the SEM with little additional preparation. It could be that the different physics of the SEM render the rings more visible.

I do suppose you would be using a variable pressure SEM. That should easily permit you to view the samples without coating.

I would consider a better surface finish. I know I can lose a lot of features in a rough surface. I can't tell how well sanded the current sample is. The dividers are certainly not well sanded. I think you might be able to take the samples through 320-grit paper fairly easily. Maybe you can go even finer if the sandpaper stays clear. Normally water would be used as a lubricant and flush for such fine papers, but that might not be feasible with the wood. A better finish might render the rings visible to the naked eye or to a hand lens.


My original response, FWIW:
Like most samples, it will depend on what they want to see.

I am materials scientist, not a biologist. I have plenty of clients come in asking to do an analysis a particular way. When I ask them what they really want to know, it often happens that their choice of method was quite inferior to other options available to them.

If these folks are looking for distribution of minerals, I would recommend embedding, sectioning and polishing. It is quite hard to pick out the minerals and get a good x-ray signal when the particles are tucked away in the nooks and crannies of a fracture surface.

Beyond that, I recommend you discuss the matter with them.

Warren S.

-----Original Message-----
X-from: rintugr-at-gmail.com [mailto:rintugr-at-gmail.com]
Sent: Wednesday, October 06, 2010 2:24 PM
To: wesaia-at-iastate.edu

Dear Soumitra,

I think it might be worth a try to put the samples in an SEM, but frankly, I
would not hold my breath. I am not a biologist, and I assume that the rings
represent wood that is slightly denser or with a slightly different
morphology, but that there are no significant changes in the overall
composition. If that is right, I would guess that there is not enough
difference for the rings to show up in BSE, and if you use a highly polished
material, which would probably have to be coated with a conductor, the SE
will likely not give you much information either.

My guess is that an optical method with stained samples would be an easier
approach. Perhaps even a simple wood stain from your neighborhood building
supply store could do the trick. How that affects mass spectrometry
afterwards, however, I don't know.

Mike Bode
---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com





-----Original Message-----
X-from: rintugr-at-gmail.com [mailto:rintugr-at-gmail.com]
Sent: Wednesday, October 06, 2010 1:35 PM
To: mike.bode-at-resaltatech.com

Dear Colleagues,

A post doctoral fellow in my department has some wood samples for
imaging (surface only, no sectioning). I was thinking of doing SEM of
a highly polished sample to see the individual rings and the cell
structure. However, it is always best to ask the experts for the right
approach. They have a large number of samples and any help in this
matter will be very much appreciated.

Given below is her email message and the dropbox link has the image of
the wooden planchets.

Thank you,

Soumitra

************************************************************
Soumitra Ghoshroy
Director, Electron Microscopy Center
Research Associate Professor, Biology
University of South Carolina
Columbia, SC 29208
803-777-7085 (office)
http://www.emc.sc.edu
************************************************************

Dear Soumitra,
my colleagues and I would like to ask about any techniques to make
birch year rings visible.
We have wood samples taken from trees growing in different ecological
conditions and we need to measure year rings. We did this with pine
samples, but it is really difficult to find year rings in birch wood
samples. We have a guess, that in is possible to color them in
someway. The samples are putted into wooden planchettes and fixed
there with glue. We need these samples for future mass-spectrometry
analysis, so the task is to make year rings visible but not destroy
samples.

Sample lengths is about 20-30 centimeters, they are air-dry, photo is added.

http://dl.dropbox.com/u/7737489/Wood_Cores.pdf

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9, 30 -- Subject: Wood microscopy
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==============================Original Headers==============================
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From: brobertson-at-unl.edu
Date: Wed, 6 Oct 2010 16:15:31 -0500
Subject: [Microscopy] Wood microscopy

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

I just read all the way to the bottom of the message.
How can you get a 20-30 cm sample into most available SEMs? If they are cut, one might loose a ring each time.
I was going to suggest cutting a sliver from the side of the board/rod and staining it like Mike suggested then viewing under a good dissecting microscope. Use the remaining part for other procedures. As far as I know the whole board could not be used for mass-spectrometry analysis either.
Why glued down in the first place? How wide are the boards? It could be 1cm if they are 20cm long.
Pat


Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E - Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not represent the NIH. This message is not confidential and can be freely shared and reproduced.

________________________________
X-from: {wesaia-at-iastate.edu}
Reply-To: {wesaia-at-iastate.edu}

Can I suggest that your colleagues consult the Tree Ring lab at the
University of AZ (where, I am told, the science of dendrochronology was
born).

See: http://www.ltrr.arizona.edu/

Best Regards,
Doug


^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Douglas W. Cromey, M.S. - Assistant Scientific Investigator
Dept. of Cell Biology & Anatomy, University of Arizona
1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA

office: AHSC 4212 email: Cromey-at-Arizona.edu
voice: 520-626-2824 fax: 520-626-2097

http://swehsc.pharmacy.arizona.edu/exppath/
Home of: "Microscopy and Imaging Resources on the WWW"


-----Original Message-----
X-from: connellyps-at-nhlbi.nih.gov [mailto:connellyps-at-nhlbi.nih.gov]
Sent: Wednesday, October 06, 2010 1:53 PM
To: dcromey-at-email.arizona.edu

I just read all the way to the bottom of the message.
How can you get a 20-30 cm sample into most available SEMs? If they are
cut, one might loose a ring each time.
I was going to suggest cutting a sliver from the side of the board/rod and
staining it like Mike suggested then viewing under a good dissecting
microscope. Use the remaining part for other procedures. As far as I know
the whole board could not be used for mass-spectrometry analysis either.
Why glued down in the first place? How wide are the boards? It could be
1cm if they are 20cm long.
Pat


Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E - Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and can be freely shared
and reproduced.

________________________________
X-from: {wesaia-at-iastate.edu}
Reply-To: {wesaia-at-iastate.edu}

Dear Mike and Soumitra,

Wood and cork have been very successfully examined in SEM after cutting
(!) with a truly sharp blade to reveal the tree rings and cellular
structure. Each of the longitudinal, tangential and radial directions of
the wood can be imaged this way. The blade must be sharp and must be
used with precision in order to reveal the essentially underformed
internal structure by cutting to remove a thin surface layer; if the
blade is not sharp enough or is not used to cut, the cell structure and
ring structure are likely to be crushed and may not be as useful to image.

The SEM work of Mike Ashby and Ken Easterling and their colleagues from
at least the 1980s on wood and cork is quite wonderful and, as far as I
know, did not involve low vacuum SEM or ESEM or embedding or polishing.
See, for example,
M. F. Ashby, K. E. Easterling, R. Harrysson and S. K. Maiti, "The
Fracture and Toughness of Woods", Proc. Roy. Soc. Lond. A, 9 April
1985, vol. 398 no. 1815 pp. 261-280 or one of the articles (at least
as recent as 2008) that cite this paper.

Best wishes,
Brian Robertson

--
Full contact information:

Professor Brian W. Robertson
Department of Mechanical Engineering
and Nebraska Center for Materials and Nanoscience
University of Nebraska-Lincoln, N124 WSEC,
17th & Vine Streets, Lincoln, NE 68588-0656, U.S.A.

http://www.engineering.unl.edu/academicunits/MechanicalEngineering/faculty-staff/BrianRobertson.shtml
http://www.unl.edu/ncmn/faculty/robertson.shtml
Office 402 472 8308
Labs 402 472-8762 and 402 472-5498
FAX 402 472 1465
--

On 10/6/2010 3:32 PM, mike.bode-at-resaltatech.com wrote:
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} Dear Soumitra,
}
} I think it might be worth a try to put the samples in an SEM, but frankly, I
} would not hold my breath. I am not a biologist, and I assume that the rings
} represent wood that is slightly denser or with a slightly different
} morphology, but that there are no significant changes in the overall
} composition. If that is right, I would guess that there is not enough
} difference for the rings to show up in BSE, and if you use a highly polished
} material, which would probably have to be coated with a conductor, the SE
} will likely not give you much information either.
}
} My guess is that an optical method with stained samples would be an easier
} approach. Perhaps even a simple wood stain from your neighborhood building
} supply store could do the trick. How that affects mass spectrometry
} afterwards, however, I don't know.
}
} Mike Bode
} ---
} Mike Bode, Ph.D.
} ResAlta Research Technologies Corp.
}
} 2102 Beech Ct
} Golden, CO 80401
} USA
}
} p: +1-303-748-4346
} f: +1-303-202-6350
} Mike.Bode-at-ResAltaTech.com
} www.ResAltaTech.com
}
}
}
}
}
} -----Original Message-----
} X-from: rintugr-at-gmail.com [mailto:rintugr-at-gmail.com]
} Sent: Wednesday, October 06, 2010 1:35 PM
} To: mike.bode-at-resaltatech.com
} Subject: [Microscopy] Wood microscopy
}
}
}
}
} ----------------------------------------------------------------------------
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} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear Colleagues,
}
} A post doctoral fellow in my department has some wood samples for
} imaging (surface only, no sectioning). I was thinking of doing SEM of
} a highly polished sample to see the individual rings and the cell
} structure. However, it is always best to ask the experts for the right
} approach. They have a large number of samples and any help in this
} matter will be very much appreciated.
}
} Given below is her email message and the dropbox link has the image of
} the wooden planchets.
}
} Thank you,
}
} Soumitra
}
} ************************************************************
} Soumitra Ghoshroy
} Director, Electron Microscopy Center
} Research Associate Professor, Biology
} University of South Carolina
} Columbia, SC 29208
} 803-777-7085 (office)
} http://www.emc.sc.edu
} ************************************************************
}
} Dear Soumitra,
} my colleagues and I would like to ask about any techniques to make
} birch year rings visible.
} We have wood samples taken from trees growing in different ecological
} conditions and we need to measure year rings. We did this with pine
} samples, but it is really difficult to find year rings in birch wood
} samples. We have a guess, that in is possible to color them in
} someway. The samples are putted into wooden planchettes and fixed
} there with glue. We need these samples for future mass-spectrometry
} analysis, so the task is to make year rings visible but not destroy
} samples.
}
} Sample lengths is about 20-30 centimeters, they are air-dry, photo is added.
}
} http://dl.dropbox.com/u/7737489/Wood_Cores.pdf
}
} ==============================Original Headers==============================
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} 9, 30 -- Subject: Wood microscopy
} 9, 30 -- From: Soumitra Ghoshroy {rintugr-at-gmail.com}
} 9, 30 -- To: Microscopy-at-microscopy.com
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} ==============================Original Headers==============================
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8, 19 -- Date: Wed, 06 Oct 2010 16:16:39 -0500
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From: rosemary.white-at-csiro.au
Date: Wed, 6 Oct 2010 16:49:05 -0500
Subject: [Microscopy] Re: Wood microscopy

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

One way to see details of the rings in SEM without processing the sample or
coating it is to use BSE in low-vacuum mode. This is a very good way of
seeing plant cell walls in material like wood without much cell contents.
We use this quite a bit for cell size measurements in cleared or
semi-cleared tissues after CPD. Older tissue doesn't need clearing as it
has less cytoplasmic contents - you see just the cell walls and nucleus (and
chloroplasts, if present).

Our SEM won't take such large samples, they may have to be imaged in
segments.

Re. ms. analysis, we had a carbon dating person cut his cores into fine
segments with our venerable sledge microtome - he collected each section in
a separate tube for analysis - each section was about one tree-ring in the
smallest rings. A tedious process but apparently it worked really well.

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au


On 7/10/10 6:30 AM, "rintugr-at-gmail.com" {rintugr-at-gmail.com} wrote:

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} Dear Colleagues,
}
} A post doctoral fellow in my department has some wood samples for
} imaging (surface only, no sectioning). I was thinking of doing SEM of
} a highly polished sample to see the individual rings and the cell
} structure. However, it is always best to ask the experts for the right
} approach. They have a large number of samples and any help in this
} matter will be very much appreciated.
}
} Given below is her email message and the dropbox link has the image of
} the wooden planchets.
}
} Thank you,
}
} Soumitra
}
} ************************************************************
} Soumitra Ghoshroy
} Director, Electron Microscopy Center
} Research Associate Professor, Biology
} University of South Carolina
} Columbia, SC 29208
} 803-777-7085 (office)
} http://www.emc.sc.edu
} ************************************************************
}
} Dear Soumitra,
} my colleagues and I would like to ask about any techniques to make
} birch year rings visible.
} We have wood samples taken from trees growing in different ecological
} conditions and we need to measure year rings. We did this with pine
} samples, but it is really difficult to find year rings in birch wood
} samples. We have a guess, that in is possible to color them in
} someway. The samples are putted into wooden planchettes and fixed
} there with glue. We need these samples for future mass-spectrometry
} analysis, so the task is to make year rings visible but not destroy
} samples.
}
} Sample lengths is about 20-30 centimeters, they are air-dry, photo is added.
}
} http://dl.dropbox.com/u/7737489/Wood_Cores.pdf
}
} ==============================Original Headers==============================
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} {AANLkTikfgGLJwg5jP-n0O2HfBYSyOCtzCi_7qJ9=NTH9-at-mail.gmail.com}
} 9, 30 -- Subject: Wood microscopy
} 9, 30 -- From: Soumitra Ghoshroy {rintugr-at-gmail.com}
} 9, 30 -- To: Microscopy-at-microscopy.com
} 9, 30 -- Content-Type: text/plain; charset=UTF-8
} ==============================End of - Headers==============================



==============================Original Headers==============================
11, 39 -- From prvs=888269c6d=Rosemary.White-at-csiro.au Wed Oct 6 16:49:04 2010
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From: ph2-at-sprynet.com
Date: Wed, 6 Oct 2010 18:03:48 -0500
Subject: [Microscopy] Wood microscopy

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

1. The Growth Ring structure should be self-evident for all but the
Tropical woods due to late wood-early wood changes in cellular structure
(and density) [tropical woods lack a good definition as they have no true
growing season for the growth fairly consistent].

2. ESEM or coating is necessary and even with coating difficulties can
arise.

As for cellular structure, a clean cut with a razor blade [use new ones very
often (1-4 cuts)]) provides a good surface to view. Otherwise, looking for
things like tyloses in oak or dentate rays in pinus sp. would be difficult.

(Note: I have embedded and ground a sample or two as I would a metal or
polymer and did not like the results)

4. The big question is what particular cellular structure is the
student interested in?

X-section, Tangential, and Radial cuts are likely necessary unless one is
very adept at recognizing the wood structure in a quasi-random orientation
(e.g., someone that works with fractured specimens or charcoal).

And if the student only wants rings, then use a stereoscope.

5. I suggest the student get ahold of the following articles and then
figure out what they really want to see.


Kucera, Splitting Wood Specimens for observation in SEM, J Microsc, 142, 1,
71-77, 1986

Fromm, Lignin distribution in wood cell walls determined by TEM and
backscattered SEM techniques, J Struct Biol, 143, 77-84, 2003

Borgin, The use of the SEM for the study of weathered wood, J Microsc, 92,
1, 47-55, 1970


There are a couple of old articles in the SEM proceedings (ca. 1968-1973)
that would be useful if the U Southern Carolina library has them. And
Cotê's Atlas would be very helpful.




Tony

......................................................................
Andrew Anthony "Tony" Havics, CHMM, CIH, PE
pH2, LLC 5250 E US 36, Suite 830 Avon IN 46123 www.ph2llc.com
(317) 718-7020 off
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consultant can give you the other 10%(SM)

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distributed without this statement.




-----Original Message-----
X-from: rintugr-at-gmail.com [mailto:rintugr-at-gmail.com]
Sent: Wednesday, October 06, 2010 3:28 PM
To: ph2-at-sprynet.com

Dear Colleagues,

A post doctoral fellow in my department has some wood samples for
imaging (surface only, no sectioning). I was thinking of doing SEM of
a highly polished sample to see the individual rings and the cell
structure. However, it is always best to ask the experts for the right
approach. They have a large number of samples and any help in this
matter will be very much appreciated.

Given below is her email message and the dropbox link has the image of
the wooden planchets.

Thank you,

Soumitra

************************************************************
Soumitra Ghoshroy
Director, Electron Microscopy Center
Research Associate Professor, Biology
University of South Carolina
Columbia, SC 29208
803-777-7085 (office)
http://www.emc.sc.edu
************************************************************

Dear Soumitra,
my colleagues and I would like to ask about any techniques to make
birch year rings visible.
We have wood samples taken from trees growing in different ecological
conditions and we need to measure year rings. We did this with pine
samples, but it is really difficult to find year rings in birch wood
samples. We have a guess, that in is possible to color them in
someway. The samples are putted into wooden planchettes and fixed
there with glue. We need these samples for future mass-spectrometry
analysis, so the task is to make year rings visible but not destroy
samples.

Sample lengths is about 20-30 centimeters, they are air-dry, photo is added.

http://dl.dropbox.com/u/7737489/Wood_Cores.pdf

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9, 30 -- Subject: Wood microscopy
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==============================Original Headers==============================
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From: paul_hazelton-at-umanitoba.ca
Date: Wed, 6 Oct 2010 18:52:06 -0500
Subject: [Microscopy] Re: viaWWW: Reusing Grids

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

Andrea, et al

Because we use between 2000-3000 formvar coated grids I have followed
this thread with some interest.

About 15 years ago I considered whether it would be feasible to clean
and reuse grids. Then I did the math.

Grids cost $21.50/100, or about 21.5c/grid. This is all you save.
Balance that against your time, the cost of reagents, and the risk that
you may not get the grids clean. The ultimate result could be that you
have to re-prepare, or even lose important specimens.

It's really not worth it.

If you want to talk sometime privately about preparation methods give me
a call. We have a very low failure rate (below 1%) for formvar coated
grids.

Paul


--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 Bannatyne Avenue
Winnipeg, Manitoba, Canada, R3E 3J7
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926


==============================Original Headers==============================
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From: tbogea-at-interchange.ubc.ca
Date: Wed, 6 Oct 2010 19:04:22 -0500
Subject: [Microscopy] Block trimer

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

Hi all

Has anyone used the Butler Block Trimer from EMS? Is it any good or just another gadget?
Thanks for sharing your experience.

Tami
--
Tami Bogea PhD
Research Associate
Ophthalmology & Visual Sciences
University of British Columbia
2550 Willow St. Vancouver, BC
V5Z 3N9 Canada


==============================Original Headers==============================
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From: nizets2-at-yahoo.com
Date: Thu, 7 Oct 2010 03:43:27 -0500
Subject: [Microscopy] Wood microscopy

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

I am not dendrochronologist but using a SEM for such a large sample with
macrostructures seems an overkill to me. Are the growth rings µm apart?
Supposing that the rings are made of the same substance than the rest, just more
dense (compressed), perhaps an easy and nice solution would be to use different
wavelengthes to illuminate the wood.
Especially in the UV range there is possibility that some molecules light up, or
fluorescence, and if they are more dense the ring will inevitably come out.
Just my 2 eurocents (which is still more worth than 2 dollar cents ;-))

Regards,
Stephane

PS: this discussion is interesting but I would contact specialists, they
definitely know better



----- Original Message ----
X-from: "rintugr-at-gmail.com" {rintugr-at-gmail.com}
To: nizets2-at-yahoo.com
Sent: Wed, October 6, 2010 9:27:40 PM

Dear Colleagues,

A post doctoral fellow in my department has some wood samples for
imaging (surface only, no sectioning). I was thinking of doing SEM of
a highly polished sample to see the individual rings and the cell
structure. However, it is always best to ask the experts for the right
approach. They have a large number of samples and any help in this
matter will be very much appreciated.

Given below is her email message and the dropbox link has the image of
the wooden planchets.

Thank you,

Soumitra

************************************************************
Soumitra Ghoshroy
Director, Electron Microscopy Center
Research Associate Professor, Biology
University of South Carolina
Columbia, SC 29208
803-777-7085 (office)
http://www.emc.sc.edu
************************************************************

Dear Soumitra,
my colleagues and I would like to ask about any techniques to make
birch year rings visible.
We have wood samples taken from trees growing in different ecological
conditions and we need to measure year rings. We did this with pine
samples, but it is really difficult to find year rings in birch wood
samples. We have a guess, that in is possible to color them in
someway. The samples are putted into wooden planchettes and fixed
there with glue. We need these samples for future mass-spectrometry
analysis, so the task is to make year rings visible but not destroy
samples.

Sample lengths is about 20-30 centimeters, they are air-dry, photo is added.

http://dl.dropbox.com/u/7737489/Wood_Cores.pdf

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9, 30 -- Subject: Wood microscopy
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9, 30 -- To: Microscopy-at-microscopy.com
9, 30 -- Content-Type: text/plain; charset=UTF-8
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24, 35 -- Date: Thu, 7 Oct 2010 01:43:24 -0700 (PDT)
24, 35 -- From: Stephane Nizet {nizets2-at-yahoo.com}
24, 35 -- Subject: Re: [Microscopy] Wood microscopy
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From: christian.liebig-at-medizin.uni-tuebingen.de
Date: Thu, 7 Oct 2010 03:51:38 -0500
Subject: [Microscopy] Re: Wood microscopy

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

Hi,

in my previous job we had a user working with Lignin autofluorescence so
I had the idea of just putting your samples under a normal fluorescence
microscope.
Googling for "lignin autofluorescence year rings" I also found this work:
ftp://ftp.wsl.ch/pub/gaertner/Trace_Volumes/Vol_6_PDF/Vavrcik_et_al_TraceVol_6.pdf

HTH, kind regards,

Christian




Am 06.10.2010 21:34, schrieb rintugr-at-gmail.com:
} ----------------------------------------------------------------------------
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} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear Colleagues,
}
} A post doctoral fellow in my department has some wood samples for
} imaging (surface only, no sectioning). I was thinking of doing SEM of
} a highly polished sample to see the individual rings and the cell
} structure. However, it is always best to ask the experts for the right
} approach. They have a large number of samples and any help in this
} matter will be very much appreciated.
}
} Given below is her email message and the dropbox link has the image of
} the wooden planchets.
}
} Thank you,
}
} Soumitra
}
} ************************************************************
} Soumitra Ghoshroy
} Director, Electron Microscopy Center
} Research Associate Professor, Biology
} University of South Carolina
} Columbia, SC 29208
} 803-777-7085 (office)
} http://www.emc.sc.edu
} ************************************************************
}
} Dear Soumitra,
} my colleagues and I would like to ask about any techniques to make
} birch year rings visible.
} We have wood samples taken from trees growing in different ecological
} conditions and we need to measure year rings. We did this with pine
} samples, but it is really difficult to find year rings in birch wood
} samples. We have a guess, that in is possible to color them in
} someway. The samples are putted into wooden planchettes and fixed
} there with glue. We need these samples for future mass-spectrometry
} analysis, so the task is to make year rings visible but not destroy
} samples.
}
} Sample lengths is about 20-30 centimeters, they are air-dry, photo is added.
}
} http://dl.dropbox.com/u/7737489/Wood_Cores.pdf
}
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} 9, 30 -- Message-ID: {AANLkTikfgGLJwg5jP-n0O2HfBYSyOCtzCi_7qJ9=NTH9-at-mail.gmail.com}
} 9, 30 -- Subject: Wood microscopy
} 9, 30 -- From: Soumitra Ghoshroy {rintugr-at-gmail.com}
} 9, 30 -- To: Microscopy-at-microscopy.com
} 9, 30 -- Content-Type: text/plain; charset=UTF-8
} ==============================End of - Headers==============================

--
Christian Liebig, PhD
Hertie-Institut für klinische Hirnforschung
Otfried-Müller-Straße 27
72076 Tübingen
Germany

Phone: ++49-7071-29-87607
Fax: ++49-7071-29-4521
E-Mail: christian.liebig-at-medizin.uni-tuebingen.de

==============================Original Headers==============================
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From: oshel1pe-at-cmich.edu
Date: Thu, 7 Oct 2010 11:08:44 -0500
Subject: [Microscopy] Re: Mystery microscope

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

Or perhaps in this case believing is seeing ...

Phil

} Dear Confocal Listers,
}
} I suggest that everyone try to get a demo of it in your labs, it
} would be interesting if they even would do such a thing. But then
} and only then can you truly know for sure whether it is a load of
} crock or not. As the old adage goes: Seeing is believing...
}
} Pete
}
} P.S. If they start to get protective of this technology and won't
} let you demo it, then you know that they are the swindlers that they
} seem to be.
}
} On Oct 7, 2010, at 14:49 PM, Robert Fernandez wrote:
}
} } *****
} } To join, leave or search the confocal microscopy listserv, go to:
} } http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
} } *****
} }
} } HI All
} }
} } Here is some more information about the mystery microscope, all
} } seems a bit strange to me but mildly entertaining too!
} }
} } http://www.rife.de/index.html
} }
} } Thanks
} }
} } Robert
} }
} } Robert Fernandez
} } Bioimaging Facility
} } University of Manchester
} } UK
} }
} } -----Original Message-----
} } From: Confocal Microscopy List
} } [mailto:CONFOCALMICROSCOPY-at-LISTS.UMN.EDU] On Behalf Of Andreas
} } Bruckbauer
} } Sent: 07 October 2010 11:30
} } To: CONFOCALMICROSCOPY-at-LISTS.UMN.EDU
} } Subject: Re: Mystery microscope
} }
} } *****
} } To join, leave or search the confocal microscopy listserv, go to:
} } http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
} } *****
} }
} }
} } I agree, seems to be some pseudoscience, the image of the
} } Richardson test slide on their webpage shows that the resolution is
} } somewhere between 260 - 300 nm.
} }
} } Andreas
} }
} }
} }
} }
} }
} }
} }
} } -----Original Message-----
} } From: Guy Cox {guy.cox-at-SYDNEY.EDU.AU}
} } To: CONFOCALMICROSCOPY-at-LISTS.UMN.EDU
} } Sent: Wed, 6 Oct 2010 17:11
} } Subject: Re: Mystery microscope
} }
} }
} } *****
} }
} } To join, leave or search the confocal microscopy listserv, go to:
} }
} } http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
} }
} } *****
} }
} }
} }
} } OK, since I was asked to comment. Nothing in this makes sense. Why
} }
} } should the 100nm resolution version cost so much more? Maybe just so
} }
} } that people don't buy it and so won't find out that it doesn't work?
} }
} }
} }
} } There are several 100nm (or better) optical resolution microscopes you
} }
} } can buy based on well published and dependable principles. (Structured
} }
} } illumination, STED, 4pi, stochastic single molecule imaging.)
} }
} }
} }
} } There are also microscopes with extended depth of field (but no lateral
} }
} } super-resolution). Carol Cogswell was among those working in this
} }
} } field.
} }
} }
} }
} } Guy
} }
} }
} }
} } Optical Imaging Techniques in Cell Biology
} }
} } by Guy Cox CRC Press / Taylor & Francis
} }
} } http://www.guycox.com/optical.htm
} }
} } ______________________________________________
} }
} } Associate Professor Guy Cox, MA, DPhil(Oxon)
} }
} } Australian Centre for Microscopy & Microanalysis,
} }
} } Madsen Building F09, University of Sydney, NSW 2006
} }
} }
} }
} } Phone +61 2 9351 3176 Fax +61 2 9351 7682
} }
} } Mobile 0413 281 861
} }
} } ______________________________________________
} }
} } http://www.guycox.net
} }
} }
} }
} }
} }
} }
} }
} } -----Original Message-----
} }
} } From: Confocal Microscopy List [mailto:CONFOCALMICROSCOPY-at-LISTS.UMN.EDU]
} }
} } On Behalf Of Martin Wessendorf
} }
} } Sent: Wednesday, 6 October 2010 1:08 AM
} }
} } To: CONFOCALMICROSCOPY-at-LISTS.UMN.EDU
} }
} } Subject: Re: Mystery microscope
} }
} }
} }
} } *****
} }
} } To join, leave or search the confocal microscopy listserv, go to:
} }
} } http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
} }
} } *****
} }
} }
} }
} } On 10/5/2010 12:49 AM, Andreas Bruckbauer wrote:
} }
} }
} }
} } } Another mystery microscope:
} }
} } } http://www.grayfieldoptical.com/
} }
} }
} }
} } Does anybody (--Guy? Barbara? Jim?) have an idea of the optical
} }
} } principles underlying this instrument? Has this been published
} }
} } anywhere?
} }
} }
} }
} } These are surprising claims (e.g. 100 nm resolution in what appears to
} }
} } be a widefield microscope)...though lately, lord knows, there've been
} }
} } lots of surprises to be had.
} }
} }
} }
} } Thanks--
} }
} }
} }
} } Martin
} }
} } --
} }
} } Martin Wessendorf, Ph.D. office: (612) 626-0145
} }
} } Assoc Prof, Dept Neuroscience lab: (612) 624-2991
} }
} } University of Minnesota Preferred FAX: (612) 624-8118
} }
} } 6-145 Jackson Hall, 321 Church St. SE Dept Fax: (612) 626-5009
} }
} } Minneapolis, MN 55455 e-mail: martinw-at-umn.edu

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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From: ebkovacs-at-shaw.ca
Date: Thu, 7 Oct 2010 12:51:46 -0500
Subject: [Microscopy] LM - Grayfield Optical Microscope

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

I have read the posts for high resolution light microscopy with interest.
This is not entirely a new concept. Royal Raymond Rife in the 1930's
designed a high resolution, high magnification light microscope that clashed
with the accepted concepts for the magnification range for light microscopes
at that time.

Leading up to the Grayfield Optical Microscope, when I managed a microscopy
lab, I happened to notice a marked improvement in my sample images when I
implemented grazing illumination across the sample from both sides. Being a
busy lab I did not follow through on this observation, however, I did run
into this same phenomena later at an International Microscopy Conference in
the early 1990's where Gary Greenberg was giving a talk about the new high
resolution Edge Microscope that he had designed. His company no longer
exists. I am not a physicist, so I am not going to attempt a detailed
explanation, but in essence what Gary said was that when light on the
specimen is unidirectional only a portion of the light information is
displayed for the sample. Adding additional lighting from the opposite sides
and above and below fills in the gaps to generate a higher resolution image.
This multi directional lighting may be observed in the Grayfield Optical
Microscope. Formation of holograms may explain the increased depth of field.

X-from my perspective, Kurt Olbrich's resolution improvement with his
instrument is at the very least impressive and opens new light microscopy
capabilities. I think it would benefit the light microscopy community to
help build on Kurt Olbrich's accomplishments.

Regards.

Barbara Kovacs
Live and Dried Blood Analyst
5982 Cobblestone Street
Chilliwack, BC, V2R 0E4
Canada
E-mail: ebkovacs-at-shaw.ca


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From: mrisley-at-nmt.edu
Date: Thu, 7 Oct 2010 23:51:55 -0500
Subject: [Microscopy] SEM - Clay mineral imaging and EDS

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

Evening Colleagues,
Does anyone have experience with SEM imaging and x-ray
analysis on clay minerals? I'm conducting research on catalyst deposition
of 2:1 smectite clays and need to characterize these clays by means of
visual inspection as well as quantitative compositional information.

I'm working with powdered clay samples, and in the past have done imaging
with no sample preparation (sputtering or anything else), although the
clays exhibit mild to moderate charging. In addition to this I have
carried out EDS on these powders which provides results with a relatively
large margin of error against standards. Does anyone have any advice or
references for sample preparation of these clays; specifically for high
resolution imaging and quantitative EDS analysis/mapping.

Thank you,

Mason Risley
Chemical Engineering Dept.
New Mexico Inst. of Mining and Technology

==============================Original Headers==============================
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From: rintugr-at-gmail.com
Date: Fri, 8 Oct 2010 07:38:19 -0500
Subject: [Microscopy] Re: Wood microscopy

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

Dear Colleagues,

Thank you so very much.

I received so many excellent responses. We will certainly try as many
of you suggested, SEM or light microscopy with strategic staining and
other methods. The wood specimen strips were collected in the past and
glued to a large piece of plywood. Therefore we will have to work with
whatever we have. I know they are too long for SEM stage, however, the
scientists are reluctant to cut it to fit the stage. We will certainly
try the light microscopy methods.

As we have a number of great procedures suggested by so many of you
and I am confident the project will be successful.

Have a wonderful weekend.

Soumitra

************************************************************
Soumitra Ghoshroy
Director, Electron Microscopy Center
Research Associate Professor, Biology
University of South Carolina
Columbia, SC 29208
803-777-7085 (office)
http://www.emc.sc.edu
************************************************************

On Wed, Oct 6, 2010 at 3:32 PM, {rintugr-at-gmail.com} wrote:
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor:  The Microscopy Society of America
} To  Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear Colleagues,
}
} A post doctoral fellow in my department has some wood samples for
} imaging (surface only, no sectioning). I was thinking of doing SEM of
} a highly polished sample to see the individual rings and the cell
} structure. However, it is always best to ask the experts for the right
} approach. They have a large number of samples and any help in this
} matter will be very much appreciated.
}
} Given below is her email message and the dropbox link has the image of
} the wooden planchets.
}
} Thank you,
}
} Soumitra
}
} ************************************************************
} Soumitra Ghoshroy
} Director, Electron Microscopy Center
} Research Associate Professor, Biology
} University of South Carolina
} Columbia, SC 29208
} 803-777-7085 (office)
} http://www.emc.sc.edu
} ************************************************************
}
} Dear Soumitra,
} my colleagues and I would like to ask about any techniques to make
} birch year rings visible.
} We have wood samples taken from trees growing in different ecological
} conditions and we need to measure year rings. We did this with pine
} samples, but it is really difficult to find year rings in birch wood
} samples. We have a guess, that in is possible to color them in
} someway. The samples are putted into wooden planchettes and fixed
} there with glue. We need these samples for future mass-spectrometry
} analysis, so the task is to make year rings visible but not destroy
} samples.
}
} Sample lengths is about 20-30 centimeters, they are air-dry, photo is added.
}
} http://dl.dropbox.com/u/7737489/Wood_Cores.pdf
}
} ==============================Original Headers==============================
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} 9, 30 -- Subject: Wood microscopy
} 9, 30 -- From: Soumitra Ghoshroy {rintugr-at-gmail.com}
} 9, 30 -- To: Microscopy-at-microscopy.com
} 9, 30 -- Content-Type: text/plain; charset=UTF-8
} ==============================End of - Headers==============================
}


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From: MSHERWOOD-at-PARTNERS.ORG
Date: Fri, 8 Oct 2010 15:10:13 -0500
Subject: [Microscopy] Re: ImmunoEM

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

To all:

I am starting an ImmunoEM project for one of our investigators. I need a
protocol for using a gold-labelled antibody that he will provide for me. (Is
this possible?) I have done a preliminary search of the literature, and I can
only find protocols that have you add the antibody and then incubate with the
gold particles (either pre-embedding or post-embedding).

I'm sure there are people much more experienced with doing this and would
appreciate any help you can give me.

Thanks in advance!
Peggy
P.S. Have a nice (long) weekend!

Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDR 214)
Massachusetts General Hospital
55 Fruit Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood-at-partners.org




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From: PhillipsT-at-missouri.edu
Date: Fri, 8 Oct 2010 15:22:28 -0500
Subject: [Microscopy] Re: ImmunoEM

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

Using a primary antibody conjugated to gold will reduce you reactivity in most cases but it is worth a try. Simply incubate for 1-24 hrs, rinse and examine. One "fun" alternative might be to stain with the primary-gold conjugate then a second gold particle of a different size conjugated to a secondary antibody against the primary species. If you primary was conjugated to 15 nm, and your secondary 6 nm, you might get a distinctive cluster of 6 nm particles surrounding a 15 nm particle. This is probably not needed since the 15 nm particle should jump out at you with a cluster. But the loss of sensitivity of having the primary conjugated to gold is going to make life tough unless you have a ton of target or it is really resistant to fixation and embedding. Lots of luck.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: MSHERWOOD-at-PARTNERS.ORG [mailto:MSHERWOOD-at-PARTNERS.ORG]
Sent: Friday, October 08, 2010 3:11 PM
To: Phillips, Thomas E.

To all:

I am starting an ImmunoEM project for one of our investigators. I need a
protocol for using a gold-labelled antibody that he will provide for me. (Is
this possible?) I have done a preliminary search of the literature, and I can
only find protocols that have you add the antibody and then incubate with the
gold particles (either pre-embedding or post-embedding).

I'm sure there are people much more experienced with doing this and would
appreciate any help you can give me.

Thanks in advance!
Peggy
P.S. Have a nice (long) weekend!

Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDR 214)
Massachusetts General Hospital
55 Fruit Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood-at-partners.org




The information in this e-mail is intended only for the person to whom it is
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From: MSHERWOOD-at-PARTNERS.ORG
Date: Fri, 8 Oct 2010 15:34:46 -0500
Subject: [Microscopy] Re: ImmunoEM

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

Since I am not that familiar with this process, I am a bit confused as well!
The protocols that I have found due mention incubating with a primary antibody,
followed by a gold-labelled secondary antibody. I realize now that that is the
way to go. As Dr. (Thomas) Philips mentioned, labelling the primary antibody
would result in a loss of sensitivity of the reaction.

Are there protocols that use primary antibodies conjugated to gold? If I follow
the established route (protocol), do you recommend the pre-embedding or
post-embedding protocol?

Again, I apologize for the confusion.

Thanks!
Peggy


Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDR 214)
Massachusetts General Hospital
55 Fruit Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood-at-partners.org




The information in this e-mail is intended only for the person to whom it is
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but does not contain patient information, please contact the sender and properly
dispose of the e-mail.



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From: PhillipsT-at-missouri.edu
Date: Fri, 8 Oct 2010 15:44:03 -0500
Subject: [Microscopy] Re: ImmunoEM

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

Pre-embedding (assuming you are labeling a surface antigen) is almost always superior to post-embedding - higher labeling, less background. But it is hard to permeabilize enough to get the relatively big gold in without damaging your morphology so if you have an intracellular antigen, post-embedding might be preferable.

It takes a ton of antibody to conjugate to gold so most people don't want to conjugate primaries to gold and lose so much of the primary. Plus you lose sensitivity during staining - a sandwich technique (primary followed by labeled secondary) is almost always superior even when you have a fluorescent labeled secondary. You can get multiple secondary labels on a single primary to increase the signal.

I doubt you will find many published examples of gold conjugated primaries. You can buy gold conjugated lectins and this would be more analogous to what you are trying to do since the lectin takes the place of a primary antibody. Gold-lectin probes work but lectins are relatively cheap and therefore can be conjugated easily without worrying about cost. And lectins recognize high abundance targets that are relatively less sensitive to loss of reactivity after fixation in comparison to proteins.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: MSHERWOOD-at-PARTNERS.ORG [mailto:MSHERWOOD-at-PARTNERS.ORG]
Sent: Friday, October 08, 2010 3:35 PM
To: Phillips, Thomas E.

Since I am not that familiar with this process, I am a bit confused as well!
The protocols that I have found due mention incubating with a primary antibody,
followed by a gold-labelled secondary antibody. I realize now that that is the
way to go. As Dr. (Thomas) Philips mentioned, labelling the primary antibody
would result in a loss of sensitivity of the reaction.

Are there protocols that use primary antibodies conjugated to gold? If I follow
the established route (protocol), do you recommend the pre-embedding or
post-embedding protocol?

Again, I apologize for the confusion.

Thanks!
Peggy


Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDR 214)
Massachusetts General Hospital
55 Fruit Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood-at-partners.org




The information in this e-mail is intended only for the person to whom it is
addressed. If you believe this e-mail was sent to you in error and the e-mail
contains patient information, please contact the Partners Compliance HelpLine at
http://www.partners.org/complianceline . If the e-mail was sent to you in error
but does not contain patient information, please contact the sender and properly
dispose of the e-mail.



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From: zaluzec-at-aaem.amc.anl.gov
Date: Sat, 9 Oct 2010 16:54:46 -0500
Subject: [Microscopy] Administrivia: Forged/Suspect Email

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Colleagues

Please ignore and immediately delete Microscopy Listserver Email from

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For safety, I have deleted this particular subscription address from the master database.
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Your Friendly Neighborhood SysOp

Nestor



===========================================
Dr. Nestor J. Zaluzec
Argonne National Laboratory
Electron Microscopy Center
Materials Science/Bldg 212
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Tel: 530-NES-TORZ (530-637-8679)
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Senior Scientist - Argonne National Laboratory
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Senior Fellow of the Computational Institute - University of Chicago
E.P. Wigner Fellow - Oak Ridge National Laboratory
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===========================================






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From: mcgeejj-at-gmail.com
Date: Sat, 9 Oct 2010 17:57:16 -0500
Subject: [Microscopy] viaWWW: Job Vacancy - Senior Electron Microscopist (Focused Ion

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Email: mcgeejj-at-gmail.com
Name: Jim McGee

Organization: BMPC-KAPL

Title-Subject: [Filtered] Job Vacancy - Senior Electron Microscopist
(Focused Ion Beam Microscopy)

Message: Knolls Atomic Power Laboratory (KAPL) is seeking qualified
candidates to fill the position of FIB Microscopist at its Niskayuna,
New York facility. The Knolls Atomic Power Laboratory is operated
for the Department of Energy by Bechtel Marine Propulsion
Corporation. KAPL employees develop advanced nuclear propulsion
technology and provide technical support for the safe and reliable
operation of existing naval reactors.

The FIB Microscopist is responsible for operating and maintaining a
dual-column Focused Ion Beam (FIB) microscope / Scanning Electron
Microscope (SEM) analytical instrument in a test support organization
to provide materials characterization and failure
analysis/problem-solving investigations of materials in response to
environmental testing and in-service performance. The FIB microscopy
work includes in-situ high resolution imaging, micro-chemical
analysis, and site-specific 3-D feature analysis, utilizing energy
dispersive spectrometry (EDS) / electron backscatter diffraction
(EBSD) characterization techniques. In addition, the individual will
utilize FIB capabilities to prepare and extract FIB-prepared samples
for TEM analysis as well as to identify / extract specific features
in samples for other microscopy and surface analytical techniques.

The FIB Microscopist will utilize their skills with dual-column FIB /
electron microscopy to evaluate material properties and provide
value-added materials characterization information to materials
development communities; perform data /image processing, including
3-D reconstruction and visualization of FIB imaging/analytical data;
analyze and interpret comprehensive and multiple datasets; and
communicate results to sponsoring groups by oral and written reports.

Selected applicants will be subjected to a federal background
investigation and must meet eligibility requirements for access to
classified matter. US citizenship is required.

You may apply for this position at :
http://jobview.monster.com/FIB-Microscopist-Job-Niskayuna-NY-US-91264728.aspx

Thanks for your interest.

Jim McGee
Lead Engineer, Chemistry Programs
BMPC-KAPL
Tel: 518-395-4612
Jim McGee {mcgeejj-at-gmail.com}

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From: ramadanhany-at-gmail.com
Date: Sun, 10 Oct 2010 10:09:12 -0500
Subject: [Microscopy] viaWWW: Image analysis software

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Email: ramadanhany-at-gmail.com
Name: Hany

Organization: University of Calgary

Title-Subject: [Filtered] Image analysis software

Message: Hi there,
I wonder what software you recommend for particle analysis on SEM
images. I am interested in getting particle size and spacing
distribution.
It does not have to be a free software, I can pay up to 500$ for a
student edition version.

Thanks a lot

Hany

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From: nyilmaz-at-mersin.edu.tr
Date: Sun, 10 Oct 2010 15:00:41 -0500
Subject: [Microscopy] removing formvar

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

Dear Members,

How can I remove formvar/carbon film from coated nickel grids in order to
perform IEM with araldite resin sections?
Thanks in advance...

Dr. Necat Yilmaz


__________ ESET NOD32 Antivirus Akýllý Güvenlik tarafýndan saðlanan bilgiler, virüs imza veritabaný sürümü: 5518 (20101009) __________

Ýleti ESET NOD32 Antivirus Akýllý Güvenlik tarafýndan denetlendi.

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From: PhillipsT-at-missouri.edu
Date: Sun, 10 Oct 2010 15:26:45 -0500
Subject: [Microscopy] removing formvar

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

Your question needs clarification. If you have picked up sections on Formvar/carbon coated films, you can simply do you immunostaining by floating the grid on a drop of the appropriate antibody solution with the section side facing the reagent. When the section is on a substrate like Formvar, you lose the option of immersion staining where you may be able to label both sides of the section but many investigators have successfully done staining of sections on Formvar grids. Formvar support is often essential for post-embedding staining of grids since it adds stability to the section. More troubling is your use of Araldite. Araldite is relatively hydrophobic and cross-linked which will both interfere with immunostaining. LR Gold, Lowicryl HM20 or K4M or other "immuno-friendly" resins may be a better choice. I am sure someone, somewhere has succeeded at least once in immunostaining Araldite but it likely reduces your chance of success. I assume the tissue is not osmicated but if it is, your chances for success are even less. Good luck.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: nyilmaz-at-mersin.edu.tr [mailto:nyilmaz-at-mersin.edu.tr]
Sent: Sunday, October 10, 2010 3:02 PM
To: Phillips, Thomas E.

Dear Members,

How can I remove formvar/carbon film from coated nickel grids in order to
perform IEM with araldite resin sections?
Thanks in advance...

Dr. Necat Yilmaz


__________ ESET NOD32 Antivirus Akýllý Güvenlik tarafýndan saðlanan bilgiler, virüs imza veritabaný sürümü: 5518 (20101009) __________

Ýleti ESET NOD32 Antivirus Akýllý Güvenlik tarafýndan denetlendi.

http://www.nod32.com.tr




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From: benada-at-biomed.cas.cz
Date: Mon, 11 Oct 2010 07:33:52 -0500
Subject: [Microscopy] High tension problem

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

Hello,
We have another problem with our Philips CM12/STEM. High tension is getting switch off about 15
seconds after it is switched on with HT panel button. I have tested it on all preset HT values
(20, 40, 60, 80, 100 and 120 kV) and it was the same for all. The hints in Troubleshooting
section of User manual did not help.
Please, did anybody have to solve such problem? Any hints welcomed.

Best regards Oldrich

--------------------------
Oldrich Benada
Institute of Microbiology, Acad. Sci. CR
Videnska 1083
142 20 Prague 4
Czech Republic

==============================Original Headers==============================
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From: protrain-at-emcourses.com
Date: Mon, 11 Oct 2010 10:00:21 -0500
Subject: [Microscopy] High tension problem

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

Hi

Your problem may be due to a number of possible problem areas. If I was
tackling the problem I would firstly check the gas pressure in the high
voltage tank. If this is fine make sure the high voltage is switched off
and extract the high voltage cable connection from the high voltage tank.
Cover the aperture with aluminium foil and wrap foil around the cable end.

Switch on the high voltage.

1. the problem has gone away - high voltage cable shorting.
2. the problem remains - high voltage tank or power supply problem.

A final check of "1" would be to take the cable to a company that deals with
high voltage equipment, x-ray sets etc. These people should be able to test
the cable and if required use your cable ends to make a new cable; probably
at only 50% of what the instrument manufacturer may charge.

Good luck

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com




-----Original Message-----
X-from: benada-at-biomed.cas.cz [mailto:benada-at-biomed.cas.cz]
Sent: 11 October 2010 13:35
To: protrain-at-emcourses.com

Hello,
We have another problem with our Philips CM12/STEM. High tension is getting
switch off about 15
seconds after it is switched on with HT panel button. I have tested it on
all preset HT values
(20, 40, 60, 80, 100 and 120 kV) and it was the same for all. The hints in
Troubleshooting
section of User manual did not help.
Please, did anybody have to solve such problem? Any hints welcomed.

Best regards Oldrich

--------------------------
Oldrich Benada
Institute of Microbiology, Acad. Sci. CR
Videnska 1083
142 20 Prague 4
Czech Republic

==============================Original Headers==============================
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+0200 (CEST)
3, 20 -- From: Oldrich Benada {benada-at-biomed.cas.cz}
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From: vakimler-at-med.wayne.edu
Date: Mon, 11 Oct 2010 19:11:16 -0500
Subject: [Microscopy] viaWWW: Expired Immunogold

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Expired Immunogold

Message: Hello,
Is there a way to reconstitute or reactivate expired immunogold
conjugates? We have EY Laboratories', Amersham Auroprobes and Janssen
immunogold dated from expiration dates of 1987-2000. Would they still
be effective after some kind of purification?

Some of these vials have never been opened and have been maintained
at the proper storage temperature.

Or, are they only useful for fiduciary markers?

Thanks,
Vickie

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From: nichols-at-agrintl.com
Date: Mon, 11 Oct 2010 19:11:49 -0500
Subject: [Microscopy] viaWWW: Jeol JSM 5800LV, sudden loss of all electronics

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Email: nichols-at-agrintl.com
Name: Neal Nichols

Organization: American Glass Research

Title-Subject: [Filtered] Jeol JSM 5800LV, sudden loss of all electronics

Message: The unit lost all displays and controls are inoperable.
There may have been an electrical interruption or surge. Vacuum is
fine. All obvious fuses OK and all power indicator lights on boards
are green. Is there a hidden reset or a likely component to check
for this failure? Any thoughts or suggestions appreciated.

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From: leunissen-at-aurion.nl
Date: Mon, 11 Oct 2010 19:56:45 -0500
Subject: [Microscopy] Re: viaWWW: Expired Immunogold

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

Hi Vickie,

Reactivation is not possible. Purification might help and sometimes conjugates are remarkably resilient, but this is more so the case for the ultra small particle range. The larger (≥ 5 nm) conjugates tend to show dissociation of the specific protein from the gold surface with time. The detached proteins would compete with the proteins on the conjugate for binding primaries and this would be your activity "loss". Your conjugates are quite old, at least 10 years but one never knows.

I would spin them down in an ultracentrifuge, collect the pellet and re-suspend in fresh buffer (TBS or PBS) with e.g, 1% BSA. That way the dissociated protein will stay in the supernatant.

If you want to follow this lead, get in touch off list and I will let you know specifics how to do this.


Good luck,


Jan Leunissen

AURION - ImmunoGold Reagents
http://www.aurion.nl


On 12/10/2010, at 1:11 PM, vakimler-at-med.wayne.edu wrote:

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} Email: vakimler-at-med.wayne.edu
} Name: Vickie Kimler
}
} Organization: Wayne State University School of Medicine
}
} Title-Subject: [Filtered] Expired Immunogold
}
} Message: Hello,
} Is there a way to reconstitute or reactivate expired immunogold
} conjugates? We have EY Laboratories', Amersham Auroprobes and Janssen
} immunogold dated from expiration dates of 1987-2000. Would they still
} be effective after some kind of purification?
}
} Some of these vials have never been opened and have been maintained
} at the proper storage temperature.
}
} Or, are they only useful for fiduciary markers?
}
} Thanks,
} Vickie
}
} Login Host: 146.9.22.18
} ---------------------------------------------------------------------------
}
} ==============================Original Headers==============================
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==============================Original Headers==============================
14, 21 -- From leunissen-at-aurion.nl Mon Oct 11 19:56:44 2010
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From: smalinskas-at-yahoo.com
Date: Tue, 12 Oct 2010 07:33:45 -0500
Subject: [Microscopy] Re: viaWWW: Jeol JSM 5800LV, sudden loss of all electronics

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

Neal,

It sounds like you had a power interruption and the unit has gone into safe mode. I hope you have the keys to your unit. To bring the unit back to life, turn the key on the right-hand POWER side of the unit to the OFF position. Wait for all relays to finish clicking. Then turn the key to ON, then momentarily to START... just like starting a car.

Stu Smalinskas
Metallurgist
SKF
Plymouth, Michigan

--- On Mon, 10/11/10, nichols-at-agrintl.com {nichols-at-agrintl.com} wrote:

} Subject: [Microscopy] viaWWW: Jeol JSM 5800LV, sudden loss of all electronics
} Name: Neal Nichols
} Organization: American Glass Research
} Title-Subject: [Filtered] Jeol JSM 5800LV, sudden loss of
} all electronics
}
} Message: The unit lost all displays and controls are
} inoperable.
} There may have been an electrical interruption or
} surge.  Vacuum is
} fine.  All obvious fuses OK and all power indicator
} lights on boards
} are green.  Is there a hidden reset or a likely
} component to check
} for this failure?  Any thoughts or suggestions
} appreciated.






==============================Original Headers==============================
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9, 29 -- Subject: Re: [Microscopy] viaWWW: Jeol JSM 5800LV, sudden loss of all electronics
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From: PhillipsT-at-missouri.edu
Date: Tue, 12 Oct 2010 07:45:06 -0500
Subject: [Microscopy] Re: viaWWW: Expired Immunogold

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

Jan is much more of an expert than I am but I am one of the most frugal scientists around so always reluctant to throw a reagent away. On the other hand, you don't want to waste a lot of time and TEM beam charges doing immunocytochemistry with reagents that are destined to fail. Here is how I test the efficacy of my gold conjugates. I use a small strip of zeta-probe charged nylon sheets but nitrocellulose or other "sticky" membranes would work. I do a series of 1 ul dots of my target. For immunogold conjugates to secondary antibodies, this means I blot rabbit or mouse IgG in various concentrations on to the membrane. After drying and blocking with BSA, I then incubate in diluted conjugated gold overnight. Ideally, you will first this as soon as you get your gold conjugates in so that you can see the threshold of detection when it is fresh and compare that to later times.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: leunissen-at-aurion.nl [mailto:leunissen-at-aurion.nl]
Sent: Monday, October 11, 2010 7:57 PM
To: Phillips, Thomas E.

Hi Vickie,

Reactivation is not possible. Purification might help and sometimes conjugates are remarkably resilient, but this is more so the case for the ultra small particle range. The larger (≥ 5 nm) conjugates tend to show dissociation of the specific protein from the gold surface with time. The detached proteins would compete with the proteins on the conjugate for binding primaries and this would be your activity "loss". Your conjugates are quite old, at least 10 years but one never knows.

I would spin them down in an ultracentrifuge, collect the pellet and re-suspend in fresh buffer (TBS or PBS) with e.g, 1% BSA. That way the dissociated protein will stay in the supernatant.

If you want to follow this lead, get in touch off list and I will let you know specifics how to do this.


Good luck,


Jan Leunissen

AURION - ImmunoGold Reagents
http://www.aurion.nl


On 12/10/2010, at 1:11 PM, vakimler-at-med.wayne.edu wrote:

}
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} ----------------------------------------------------------------------------
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} please copy both vakimler-at-med.wayne.edu as well as the MIcroscopy
} Listserver
} ---------------------------------------------------------------------------
}
} Email: vakimler-at-med.wayne.edu
} Name: Vickie Kimler
}
} Organization: Wayne State University School of Medicine
}
} Title-Subject: [Filtered] Expired Immunogold
}
} Message: Hello,
} Is there a way to reconstitute or reactivate expired immunogold
} conjugates? We have EY Laboratories', Amersham Auroprobes and Janssen
} immunogold dated from expiration dates of 1987-2000. Would they still
} be effective after some kind of purification?
}
} Some of these vials have never been opened and have been maintained
} at the proper storage temperature.
}
} Or, are they only useful for fiduciary markers?
}
} Thanks,
} Vickie
}
} Login Host: 146.9.22.18
} ---------------------------------------------------------------------------
}
} ==============================Original Headers==============================
} 9, 22 -- From zaluzec-at-microscopy.com Mon Oct 11 19:11:15 2010
} 9, 22 -- Received: from znl.com ([206.69.208.20])
} 9, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id o9C0BFls010367
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==============================Original Headers==============================
14, 21 -- From leunissen-at-aurion.nl Mon Oct 11 19:56:44 2010
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23, 39 -- From: "Phillips, Thomas E." {PhillipsT-at-missouri.edu}
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23, 39 -- Subject: RE: [Microscopy] Re: viaWWW: Expired Immunogold
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From: cpawlowicz-at-ubmtechinsights.com
Date: Tue, 12 Oct 2010 07:54:38 -0500
Subject: [Microscopy] RE: viaWWW: Image analysis software

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

Two solutions come to mind-

Image J is a pretty popular free piece of software for all kinds of things, including counting and tracking particles..

http://www.macbiophotonics.ca/imagej/particle_analysis.htm#particle_auto

Download it from here: http://rsbweb.nih.gov/ij/


Image Metrology sells a pretty powerful piece of software called 'SPIP' which has particle analysis
http://www.imagemet.com/


Chris Pawlowicz, B.Eng
Manager, Lab Development
UBM TechInsights
3000 Solandt Rd Ottawa ON Canada K2K 2X2
T: +1 613 576 0150
F: +1 613 599 6501
E: cpawlowicz-at-ubmtechinsights.com


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

Email: ramadanhany-at-gmail.com
Name: Hany

Organization: University of Calgary

Title-Subject: [Filtered] Image analysis software

Message: Hi there,
I wonder what software you recommend for particle analysis on SEM images. I am interested in getting particle size and spacing distribution.
It does not have to be a free software, I can pay up to 500$ for a student edition version.

Thanks a lot

Hany


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From: paul_hazelton-at-umanitoba.ca
Date: Tue, 12 Oct 2010 08:02:53 -0500
Subject: [Microscopy] Re: viaWWW: Expired Immunogold

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

Vickie

Back in pre-history we did not have Jan to provide us with such
wonderful products - and he would never advertise with whom he is
associated, but their gold conjugates are very good. In those dark ages
I made my own gold conjugates, and tested them just as Tom has
recommended - with one minor change. I also did a silver enhancement of
the gold to help make sure I could see it. The caution there is that
the silver may enhance unconjugated gold which has non-specifically
stuck to the nylon or nitrocellulose membrane, so you have to be careful
with your washing steps.

Paul
--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 Bannatyne Avenue
Winnipeg, Manitoba, Canada, R3E 3J7
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926


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From: Pamela.Lloyd-at-wpafb.af.mil
Date: Wed, 13 Oct 2010 05:34:50 -0500
Subject: [Microscopy] MSORV Fall Meeting

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

The Microscopy Society of the Ohio River Valley (MSORV) invites you to
attend the upcoming Fall Meeting. Please see details below along with
further details including Meeting directions, maps, parking information
available on the MSORV website, http://www.msorv.org/.

2010 Fall Meeting
October 21st 3-7pm at
Miami University, Oxford Ohio
Electron Microscopy Facility, Room 9 Upham Hall

Including Tours of Facility and Vendor Exhibits

Featuring Student Competition for Best Microscopy Presentation


Agenda - MSORV Fall Meeting
October 21, 2010

2:30-3:30PM Mixer/Reception / Tours of facility/Talk with Exhibitors-

3:30 - 3:40PM - Welcome (Matt Chestnut and Richard Edelmann)

3:40-4:55PM - "Electron Microscopy Investigation of Carbon Nanotube
Growth on Diamond Substrate" by B. T. Quinton, Wright State
University/WPAFB

3:55-4:10PM - "Multifaceted Role of Discoidin Domain Receptors in Bone
Remodeling" by Angela R. Blissett, OSU

4:25-4:40PM - "STEM Defect Analysis and Image Simulation" by Patrick
J. Phillips, OSU

4:10-4:25PM - "Imaging Superparamagnetic Nanoparticles" by Tanya Nocera,
OSU Department of Biomedical Engineering.

4:40-4:55PM "Shape Changes in Patterned InAs as a Function of Thickness
and Temperature" by M. Twyman, Wright State University.

4:55-5:10PM - "Assessing Intracardiac Flow as a Developmental Morphogen
Using Novel 4D Imaging and Microviscometry Techniques" by Michael
Craig, UC College of Medicine

5:10-5:25PM - "Multi-scale Hierarchical Interfaces to Suppress
Interfacial Delamination in Composites: by Anil Karumuri, Wright State
University

30 min break - short business meeting

5:55-6:10PM - "Visualization of cytoskeletons of Mycoplasma penetrans
and Mcyoplasma iowae" by D.A. Jurkovic, Miami University

6:10-6:25PM - "Fabrication of Highly Active Catalysts by Attachment of
Palladium Nanoparticles on Hierarchical Carbon Structures" by Hema
Vijwani, Wright State University.

6:25-6:40PM - "The Role of Frs2 in Lens Development" by BP
Madakashira, Miami University.

6:40-6:55PM - "Functionalization of Carbon Nanostructures with Silver
Nanoparticles for Biomedical Devices" by Adam Maleszewski, Wright State
University.

6:55-7:10PM - "Changes in Cerebrovascular Innervation Following Axotomy
of the Superior Cervical Ganglion."Zoe Hesp, Miami University

7:10-7:25PM - "Study of Structure and Defects of the Double Perovskite
SR2FeMoO6" by Manisha Dixit, OSU.
15 min break to decide winners

7:40PM- Awards





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From: parishcm-at-ornl.gov
Date: Wed, 13 Oct 2010 05:55:50 -0500
Subject: [Microscopy] RE: High tension problem

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

Oldrich,

I had a similar problem with a CM30 at my previous institution. I would go to the "parameters" page and then click the HT-on button. The voltage would ramp up to perhaps 25 kV and then ramp back down to 0 and then turn off the HT button, over the course of several seconds.

Our local service engineer spent several days trying to find an easy fix to the problem, such as a bad HT cable or a bad power supply, but the solution ended up being a replacement HT tank.

--Chad Parish


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5, 34 -- Date: Wed, 13 Oct 2010 06:55:49 -0400
5, 34 -- From: "Parish, Chad M." {parishcm-at-ornl.gov}
5, 34 -- Subject: [Microscopy] RE: High tension problem
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From: nizets2-at-yahoo.com
Date: Wed, 13 Oct 2010 09:08:39 -0500
Subject: [Microscopy] RE: High tension problem

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

On our Tecnai G20, often when I start the HT it also shuts off when it reaches
between 10 and 20kV.
The solution with our instrument is very simple because we can choose the
voltage very precisely, hopefully it is also possible with your instrument:
I start the HT at 3-5kV, let it stabilize 10 seconds then I increase
progressively (but fast) up to the desired HT. It always works!

Wish you luck,

Stephane

 


----- Original Message ----
X-from: "parishcm-at-ornl.gov" {parishcm-at-ornl.gov}
To: nizets2-at-yahoo.com
Sent: Wed, October 13, 2010 12:58:50 PM

Oldrich,

I had a similar problem with a CM30 at my previous institution.  I would go to
the "parameters" page and then click the HT-on button.  The voltage would ramp
up to perhaps 25 kV and then ramp back down to 0 and then turn off the HT
button, over the course of several seconds. 


Our local service engineer spent several days trying to find an easy fix to the
problem, such as a bad HT cable or a bad power supply, but the solution ended up
being a replacement HT tank. 


--Chad Parish


==============================Original Headers==============================
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5, 34 -- Date: Wed, 13 Oct 2010 06:55:49 -0400
5, 34 -- From: "Parish, Chad M." {parishcm-at-ornl.gov}
5, 34 -- Subject: [Microscopy] RE: High tension problem
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From: jacqueline.ayotte-at-ticona.com
Date: Wed, 13 Oct 2010 09:26:09 -0500
Subject: [Microscopy] viaWWW: Image Analysis - 'quantifying' particle distribution in a

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Email: jacqueline.ayotte-at-ticona.com
Name: Jackie Ayotte

Organization: Ticona Engineering Polymers

Title-Subject: [Filtered] Image Analysis -
'quantifying' particle distribution in a matrix

Message: Hello All,
I am frequently asked to assess and show
distribution of filler in a polymer matrix.
Images show distribution qualitatively and my
customers would like something more quantitative
to be recorded, to use especially when comparing
samples.

Mostly I can take images where there is good
contrast between the filler and matrix, so Image
Analysis software (ImagePro v7) can easily detect
the filler.

What I would like to do ëquantitativelyí
Show the distance between neighboring particles (perhaps the average)
Prove/ disprove homogeneous dispersion across an image

I have not yet given this a whole lot of time ñ
why reinvent the wheel if there is a method
somewhere? I did come up with scatter plots,
which can be overlaid (but, yes, so can images),
however nothing numerical.
Perhaps I can have the software locate the
particle centers and I can put up a grid on the
image and have ImagePro measure distances between
grid lines???

A short while ago Image J was mentioned, but I
did not see anything in the software that would
quantify particle distribution across an image.

Anyway, if someone is doing this already and
would like to share some pointers I would very
much appreciate it.

If no one seems to be doing this already and I
figure out a way to get quantitative data to show
particle distribution, I'll share if anyone's
interested.

Best Regards,
Jackie


Login Host: 148.163.178.12
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From: Woody.White-at-areva.com
Date: Wed, 13 Oct 2010 09:35:40 -0500
Subject: [Microscopy] RE: High tension problem

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

Hi Stephane,

I have seen this happen when the gun area is dirty and/or the vacuum is not so good.

Woody


N.W. (Woody) White Jr.
Senior Electron Microscopist
Chemistry and Materials Center
AREVA NP Inc
An AREVA and Siemens Company
Lynchburg, VA
Lab Phone: 434.832.3004

-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Wednesday, October 13, 2010 10:22 AM
To: WHITE Norvell (RS/IB)

On our Tecnai G20, often when I start the HT it also shuts off when it reaches between 10 and 20kV.
The solution with our instrument is very simple because we can choose the voltage very precisely, hopefully it is also possible with your instrument:
I start the HT at 3-5kV, let it stabilize 10 seconds then I increase progressively (but fast) up to the desired HT. It always works!

Wish you luck,

Stephane

 


----- Original Message ----
X-from: "parishcm-at-ornl.gov" {parishcm-at-ornl.gov}
To: nizets2-at-yahoo.com
Sent: Wed, October 13, 2010 12:58:50 PM

Oldrich,

I had a similar problem with a CM30 at my previous institution.  I would go to the "parameters" page and then click the HT-on button.  The voltage would ramp up to perhaps 25 kV and then ramp back down to 0 and then turn off the HT button, over the course of several seconds. 


Our local service engineer spent several days trying to find an easy fix to the problem, such as a bad HT cable or a bad power supply, but the solution ended up being a replacement HT tank. 


--Chad Parish


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22, 35 -- Date: Wed, 13 Oct 2010 06:57:41 -0700 (PDT) 22, 35 -- From: Stephane Nizet {nizets2-at-yahoo.com} 22, 35 -- Subject: Re: [Microscopy] RE: High tension problem 22, 35 -- To: microscopy-at-microscopy.com 22, 35 -- In-Reply-To: {201010131058.o9DAwoPB009901-at-ns.microscopy.com}
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From: FMonson-at-wcupa.edu
Date: Wed, 13 Oct 2010 10:15:15 -0500
Subject: [Microscopy] RE: High tension problem

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

X-from the first day of installation of our Tecnai 12T when 'bringing up' the high tension, there has been a shut of around 60kV. The fix is to restart HT at 60kV and move on. When restarting the HT after breaking the vacuum, one is recommended to bring up the HT slowly (1kV/step - I use 3kV). Even with that, the HT stops around 60kV. 8 years with the Tecnai, and, as with my quirks, after 44 years, my wife has gotten used to it. We still have not accumulated enough to permit her to hire a divorce lawyer.

Cheers,

Fred Monson

-----Original Message-----
X-from: Woody.White-at-areva.com [mailto:Woody.White-at-areva.com]
Sent: Wednesday, October 13, 2010 10:43 AM
To: Monson, Frederick

Hi Stephane,

I have seen this happen when the gun area is dirty and/or the vacuum is not so good.

Woody


N.W. (Woody) White Jr.
Senior Electron Microscopist
Chemistry and Materials Center
AREVA NP Inc
An AREVA and Siemens Company
Lynchburg, VA
Lab Phone: 434.832.3004

-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Wednesday, October 13, 2010 10:22 AM
To: WHITE Norvell (RS/IB)

On our Tecnai G20, often when I start the HT it also shuts off when it reaches between 10 and 20kV.
The solution with our instrument is very simple because we can choose the voltage very precisely, hopefully it is also possible with your instrument:
I start the HT at 3-5kV, let it stabilize 10 seconds then I increase progressively (but fast) up to the desired HT. It always works!

Wish you luck,

Stephane

 


----- Original Message ----
X-from: "parishcm-at-ornl.gov" {parishcm-at-ornl.gov}
To: nizets2-at-yahoo.com
Sent: Wed, October 13, 2010 12:58:50 PM

Oldrich,

I had a similar problem with a CM30 at my previous institution.  I would go to the "parameters" page and then click the HT-on button.  The voltage would ramp up to perhaps 25 kV and then ramp back down to 0 and then turn off the HT button, over the course of several seconds. 


Our local service engineer spent several days trying to find an easy fix to the problem, such as a bad HT cable or a bad power supply, but the solution ended up being a replacement HT tank. 


--Chad Parish


==============================Original Headers==============================
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- Date: Wed, 13 Oct 2010 06:55:49 -0400 5, 34 -- From: "Parish, Chad M." {parishcm-at-ornl.gov} 5, 34 -- Subject: [Microscopy] RE: High tension problem 5, 34 -- To: "Microscopy-at-Microscopy.Com" {Microscopy-at-Microscopy.Com} 5, 34 -- Message-id:
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From: benada-at-biomed.cas.cz
Date: Wed, 13 Oct 2010 10:30:27 -0500
Subject: [Microscopy] Re: RE: High tension problem

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

Hi,
I would like to thank all who kindly have sent me the hints and advices, how to solve the HT
problem. Up to now I have done following but without success:

I have checked the LEDs of 24 V sources (24V fil. and 24V h.t): They were lighting for the same
time as the LED of HT button on the main panel (approx. 15 sec).

I have dismounted wehnelt assembly and checked HT.

I have replaced wehnelt assembly with the other one (freshly cleaned according to User manual
procedure, heated in the oven and mounted hot) and checked HT.

I let the scope on over night (usually it has to be in stand by) to be sure, that the vacuum is
OK (IGP = 5) and start HT at 20kV (this is the minimum value for CM12) and checked HT.

Today, the scope was in stand by over night, I have tried to switch on HT again (IGP=5) but it
did not start at all. No 15 seconds of hope as before. All the LEDs of main panel and 24V
sources did not start light.

The other suggested test are waiting for the next week.

Thank you all again.

Best regards
Oldrich

==============================Original Headers==============================
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From: bfoster-at-the-mip.com
Date: Wed, 13 Oct 2010 10:51:11 -0500
Subject: [Microscopy] Fwd: Re: viaWWW: Image Analysis - 'quantifying'

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


} Date: Wed, 13 Oct 2010 10:18:04 -0500
} To: jacqueline.ayotte-at-ticona.com, MSA
} From: Barbara Foster {bfoster-at-mme1.com}
} Subject: Re: [Microscopy] viaWWW: Image Analysis
} - 'quantifying' particle distribution in a
}
} Dear Jacqueline
}
} There is a new holographic microscopic available
} from Resolution Optics (resolutionoptics.com)
} that might do the trick. They track/measure particles in 3D.
}
} Alternatively, you can use the fine-focus on
} your microscope to take a series of pictures at
} different depths then use a sterology program to
} measure. Stereology is not to be confused with
} stereo imaging. It is a mathematical approach
} which derives 3D informaiton from 2D
} sections. Underwood is one of the big names, so
} if you can find articles or his book, that would
} be a good place to start. Also, this technique
} has been widely used by metallographers for
} decades...if you can't find one in your
} neighborhood, try getting in touch with Dr.
} Barry Fookes at U. Central Florida/Chemistry dept.
}
} Good hunting!
} Barbara Foster, President and Sr. Consultant
}
} Microscopy/Microscopy Education
} 7101 Royal Glen Trail, Suite A
} McKinney TX 75070
} P: (972)924-5310 Skype: fostermme
} W: www.MicroscopyEducation.com
}
} Working in SEM/TEM? Take part in our latest
} survey. Visit www.MicroscopyEducation.com for
} details. Survey ends Oct 18th.
}
}
}
} At 09:33 AM 10/13/2010, jacqueline.ayotte-at-ticona.com wrote:
}
}
}
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America



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4, 26 -- Subject: Fwd: Re: [Microscopy] viaWWW: Image Analysis - 'quantifying'
4, 26 -- particle distribution in a
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From: bozhilov-at-ucr.edu
Date: Wed, 13 Oct 2010 11:04:13 -0500
Subject: [Microscopy] SEAM

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

Is it possible to use the SEAM (scanning electron acoustic microscopy) method in the SEM to estimate differences in the local thermal conductivity on nanometer scale range? Also are there any established manufacturers of SEAM detectors?

Krassimir Bozhilov
Central Facility for Advanced Microscopy and Microanalysis
University of California
Riverside, CA 92521

tel. 951 827 2998
fax 951 827 2489
bozhilov-at-ucr.edu




==============================Original Headers==============================
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From: khrach-at-mediacy.com
Date: Wed, 13 Oct 2010 12:45:10 -0500
Subject: [Microscopy] Fwd: Re: viaWWW: Image Analysis - 'quantifying'

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

Hi Jackie,

I'm posting the response below on behalf of my colleague, Yuri Gaidoukevitch. He has included some details on how you can use Image-Pro Plus software for particle distribution analysis.

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

Hi Jackie,

There are several methods you can use in Image-Pro to quantify particle distribution:

1. You can create Voronoy map and measure area distribution. Use the following steps - a) Binarize background (particles - black, background - white) b) run Thinning filter to skeletonize it c) measure dark areas. The areas on the result image will correspond to the distance between particles (square root), so from the mean value you can get average distance and from standard deviation or max/min - homogeneity of the distribution.

2. You can also use Grid tool of Image-Pro to divide image into square cells and then count the number of particles in each cell (reduce particles to black dots, cells are white, and count holes). The number of holes per cell will give you similar results to method 1.


Regards,
Yuri Gaidoukevitch
Media Cybernetics

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

Kathy Hrach
Product Manager
Media Cybernetics
4340 East-West Hwy, Suite 400
Bethesda, MD 20814
Phone: 301-495-3305 ext.260
Mobile: 240-372-2010
Email: khrach-at-mediacy.com
www.mediacy.com

Attend our free Imaging Webinars:
http://www.mediacy.com/index.aspx?page=WebinarCalendar

Join our Image-Pro & AutoQuant Users Lists:
http://listserv.mediacy.com/scripts/wa.exe?INDEX


-----Original Message-----
X-from: bfoster-at-the-mip.com [mailto:bfoster-at-the-mip.com]
Sent: Wednesday, October 13, 2010 11:57 AM
To: Hrach, Kathy


} Date: Wed, 13 Oct 2010 10:18:04 -0500
} To: jacqueline.ayotte-at-ticona.com, MSA
} From: Barbara Foster {bfoster-at-mme1.com}
} Subject: Re: [Microscopy] viaWWW: Image Analysis
} - 'quantifying' particle distribution in a
}
} Dear Jacqueline
}
} There is a new holographic microscopic available
} from Resolution Optics (resolutionoptics.com)
} that might do the trick. They track/measure particles in 3D.
}
} Alternatively, you can use the fine-focus on
} your microscope to take a series of pictures at
} different depths then use a sterology program to
} measure. Stereology is not to be confused with
} stereo imaging. It is a mathematical approach
} which derives 3D informaiton from 2D
} sections. Underwood is one of the big names, so
} if you can find articles or his book, that would
} be a good place to start. Also, this technique
} has been widely used by metallographers for
} decades...if you can't find one in your
} neighborhood, try getting in touch with Dr.
} Barry Fookes at U. Central Florida/Chemistry dept.
}
} Good hunting!
} Barbara Foster, President and Sr. Consultant
}
} Microscopy/Microscopy Education
} 7101 Royal Glen Trail, Suite A
} McKinney TX 75070
} P: (972)924-5310 Skype: fostermme
} W: www.MicroscopyEducation.com
}
} Working in SEM/TEM? Take part in our latest
} survey. Visit www.MicroscopyEducation.com for
} details. Survey ends Oct 18th.
}
}
}
} At 09:33 AM 10/13/2010, jacqueline.ayotte-at-ticona.com wrote:
}
}
}
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America



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4, 26 -- Subject: Fwd: Re: [Microscopy] viaWWW: Image Analysis - 'quantifying'
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From: khrach-at-mediacy.com
Date: Wed, 13 Oct 2010 13:22:31 -0500
Subject: [Microscopy] Fwd: Re: viaWWW: Image Analysis - 'quantifying'

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

And, as another alternative, there is an Image-Pro Plus macro which can be downloaded at no charge from the Media Cybernetics Solutions Zone website. The macro is titled "Surrounding Nearest Neighbor Distances" - Solution Number 1293.

It allows measuring either center-to-center distances as well as edge-to-edge distances (using a Voronoi map).

Here's the direct link:
"Surrounding Nearest Neighbor Distances" - Solution Number 1293
http://www.mediacy.com/index.aspx?page=ViewSolution&solid=1083

Let us know if you have any questions!

Thanks,
Kathy


Kathy Hrach
Product Manager
Media Cybernetics
4340 East-West Hwy, Suite 400
Bethesda, MD 20814
Phone: 301-495-3305 ext.260
Mobile: 240-372-2010
Email: khrach-at-mediacy.com

www.mediacy.com
Attend our free Imaging Webinars:
http://www.mediacy.com/index.aspx?page=WebinarCalendar
Join our Image-Pro & AutoQuant Users Lists:
http://listserv.mediacy.com/scripts/wa.exe?INDEX


-----Original Message-----
X-from: Hrach, Kathy
Sent: Wednesday, October 13, 2010 1:44 PM
To: 'jacqueline.ayotte-at-ticona.com'; 'microscopy-at-microscopy.com'


} Date: Wed, 13 Oct 2010 10:18:04 -0500
} To: jacqueline.ayotte-at-ticona.com, MSA
} From: Barbara Foster {bfoster-at-mme1.com}
} Subject: Re: [Microscopy] viaWWW: Image Analysis
} - 'quantifying' particle distribution in a
}
} Dear Jacqueline
}
} There is a new holographic microscopic available
} from Resolution Optics (resolutionoptics.com)
} that might do the trick. They track/measure particles in 3D.
}
} Alternatively, you can use the fine-focus on
} your microscope to take a series of pictures at
} different depths then use a sterology program to
} measure. Stereology is not to be confused with
} stereo imaging. It is a mathematical approach
} which derives 3D informaiton from 2D
} sections. Underwood is one of the big names, so
} if you can find articles or his book, that would
} be a good place to start. Also, this technique
} has been widely used by metallographers for
} decades...if you can't find one in your
} neighborhood, try getting in touch with Dr.
} Barry Fookes at U. Central Florida/Chemistry dept.
}
} Good hunting!
} Barbara Foster, President and Sr. Consultant
}
} Microscopy/Microscopy Education
} 7101 Royal Glen Trail, Suite A
} McKinney TX 75070
} P: (972)924-5310 Skype: fostermme
} W: www.MicroscopyEducation.com
}
} Working in SEM/TEM? Take part in our latest
} survey. Visit www.MicroscopyEducation.com for
} details. Survey ends Oct 18th.
}
}
}
} At 09:33 AM 10/13/2010, jacqueline.ayotte-at-ticona.com wrote:
}
}
}
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America



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From: tina-at-pbrc.hawaii.edu
Date: Wed, 13 Oct 2010 14:23:26 -0500
Subject: [Microscopy] soft cells get crunchy in fixative?

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

Dear Collective-

Once in a long while I have a problem where tissue that should be soft gets
hard and crunchy during processing for TEM. Last year it was nematocysts in a
deep sea coral that were so hard that they could not be sectioned and fell out
of the sections. Some nematocysts in the tissue were fine.

This week my problem is a red alga. The green alga that I fixed at the
same time is fine. The red is too "crunchy" to section. For both the
nematocysts and the alga the one micrometer sections stain very heavily
with Richardson's stain, and the glass knife becomes scratched. The
material falls out of ultrathin sections and I feel like it's genuinely
putting some wear and tear on the diamond knife.

In both cases we "felt" like it's a reaction with fixative, but in both
cases we have no way to collect more material to compare (unless
oceanographic cruises suddenly become cheap).

There's almost no relationship between the fixatives:

The coral was fixed with 4% glutaraldehyde in 0.1M cacodylate buffer with
sucrose to raise the osmolarity, postfixed with 1% osmium tetroxide,
dehydrated with ethanol and embedded in LX112 epoxy resin. The algae were
fixed with Karnovsky's 2% formaldehyde and 5% glutaraldehyde in Sorenson's
phosphate buffer, postfixed with 1% osmium tetroxide, dehydrated with
ethanol and embedded in Spurr's original formula. Green algae looked
great, red just terrible.

Any idea why they would have gone hard and crunchy?

Aloha,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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From: oshel1pe-at-cmich.edu
Date: Wed, 13 Oct 2010 14:37:17 -0500
Subject: [Microscopy] Re: soft cells get crunchy in fixative?

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

Tina,

Can you dry some of the unfixed/unembedded algae and throw it in your
SEM/EDS? Don't worry about morphology too much, just so you can see
the cell walls.
Some red algae deposit CaCO3, and maybe that's what you're crunching up.

Phil

} Dear Collective-
}
} Once in a long while I have a problem where tissue that should be soft gets
} hard and crunchy during processing for TEM. Last year it was nematocysts in a
} deep sea coral that were so hard that they could not be sectioned and fell out
} of the sections. Some nematocysts in the tissue were fine.
}
} This week my problem is a red alga. The green alga that I fixed at the
} same time is fine. The red is too "crunchy" to section. For both the
} nematocysts and the alga the one micrometer sections stain very heavily
} with Richardson's stain, and the glass knife becomes scratched. The
} material falls out of ultrathin sections and I feel like it's genuinely
} putting some wear and tear on the diamond knife.
}
} In both cases we "felt" like it's a reaction with fixative, but in both
} cases we have no way to collect more material to compare (unless
} oceanographic cruises suddenly become cheap).
}
} There's almost no relationship between the fixatives:
}
} The coral was fixed with 4% glutaraldehyde in 0.1M cacodylate buffer with
} sucrose to raise the osmolarity, postfixed with 1% osmium tetroxide,
} dehydrated with ethanol and embedded in LX112 epoxy resin. The algae were
} fixed with Karnovsky's 2% formaldehyde and 5% glutaraldehyde in Sorenson's
} phosphate buffer, postfixed with 1% osmium tetroxide, dehydrated with
} ethanol and embedded in Spurr's original formula. Green algae looked
} great, red just terrible.
}
} Any idea why they would have gone hard and crunchy?
}
} Aloha,
} Tina
}
} ****************************************************************************
} * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
} * Biological Electron Microscope Facility * (808) 956-6251 *
} * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
} ****************************************************************************

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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5, 25 -- X-Spam-Score: -4.20 () [Hold at 6.00] L_EXCH_MF,RDNS_NONE,Bayes(0.0001:-0.5)
5, 25 -- X-CanIt-Geo: ip=141.209.15.74; country=US; region=MI; city=Mount Pleasant; postalcode=48859; latitude=43.5647; longitude=-84.8473; metrocode=513; areacode=989; http://maps.google.com/maps?q=43.5647,-84.8473&z=6
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From: susan.trant-at-viha.ca
Date: Wed, 13 Oct 2010 19:49:57 -0500
Subject: [Microscopy] viaWWW: gluteraldehyde

Contents Retrieved from Microscopy Listserver Archives
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Tina,

You may be able to rescue some of the crunchy parts by picking up the little pieces with a loupe and depositing them on a coated grid or touching a coated grid to the surface of the boat water where there is a small deposit of fragments. Some of the bits may be too thick but others may be fine.

Phil's explanation seems valid.

I agree that you should use an old diamond knife or glass for sectioning these difficult specimens.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E - Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not represent the NIH. This message is not confidential and can be freely shared and reproduced.
________________________________
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Email: susan.trant-at-viha.ca
Name: Sue Trant

Organization: Vancouver Island Health Authority

Title-Subject: [Filtered] gluteraldehyde

Message: Hello all

We currently receive our gluteraldehyde in 8% - 10ml vials. We then
buffer them down and aliquot to a 3.5 % solution for use in the EM
lab. In hopes of maximizing our resources/time we are looking into
purchasing gluteraldehyde already aliquoted and diluted. I have seen
that there are vendors that supply 2.5 % gluteraldehyde in buffers.
What is the opinion of the lower concentration and does anyone
purchase these products?


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From: ron.doole-at-materials.ox.ac.uk
Date: Thu, 14 Oct 2010 06:10:40 -0500
Subject: [Microscopy] High tension problem

Contents Retrieved from Microscopy Listserver Archives
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Hi,

We have a similar problem with our CM20. In our case it is due to the 48V HT power supplies. If we switch off the HT and unplug the two 48v power supplies (A18 and A19 in our instruemnt) until all voltages discharge (give it 3 mins) then plug them both back in. Switch on the HT and it usually works. If it does not work then in our case it is because we have not left it long enough for the power supplies to discharge so we repeat the proceedure with a little more patience. Usually the leds on these power supplies come on when the HT is switched on and go off when the HT is switched off but we have found that if the leds stay on when the HT is switched off then we will have this fault. It is something to do with how switched mode power supplies operate.

Good luck,
Ron

Mr. Ron Doole Department of Materials
Senior Instrumentation Engineer. University of Oxford.
Phone +44 (0) 1865 273701 Parks Road. Oxford. OX1 3PH
Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
________________________________________
X-from: benada-at-biomed.cas.cz [benada-at-biomed.cas.cz]
Sent: 11 October 2010 13:44
To: Ron Doole

Hello,
We have another problem with our Philips CM12/STEM. High tension is getting switch off about 15
seconds after it is switched on with HT panel button. I have tested it on all preset HT values
(20, 40, 60, 80, 100 and 120 kV) and it was the same for all. The hints in Troubleshooting
section of User manual did not help.
Please, did anybody have to solve such problem? Any hints welcomed.

Best regards Oldrich

--------------------------
Oldrich Benada
Institute of Microbiology, Acad. Sci. CR
Videnska 1083
142 20 Prague 4
Czech Republic

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From: nizets2-at-yahoo.com
Date: Thu, 14 Oct 2010 09:14:49 -0500
Subject: [Microscopy] viaWWW: gluteraldehyde

Contents Retrieved from Microscopy Listserver Archives
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Hi Susan!

We don't use glutaraldehyde alone but in combination with other stuff, like
(para)formaldeyhde, thus we always need to prepare a specific buffer and we
don't consider that diluting 10x a 25% glutaraldehyde solution increases
significantly our burden.

Regards,

Stephane



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Email: susan.trant-at-viha.ca
Name: Sue Trant

Organization: Vancouver Island Health Authority

Title-Subject: [Filtered] gluteraldehyde

Message: Hello all

We currently receive our gluteraldehyde in 8% - 10ml vials. We then
buffer them down and aliquot to a 3.5 % solution for use in the EM
lab. In hopes of maximizing our resources/time we are looking into
purchasing gluteraldehyde already aliquoted and diluted. I have seen
that there are vendors that supply 2.5 % gluteraldehyde in buffers.
What is the opinion of the lower concentration and does anyone
purchase these products?


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From: tina-at-pbrc.hawaii.edu
Date: Thu, 14 Oct 2010 13:50:38 -0500
Subject: [Microscopy] soft cells get crunchy in fixative?

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

Hi, All-

Thanks for the many replies I've received about my crunchy tissue. Most of
you have suggested ways I might decalcify and section the stuff. That's
not the problem! I do that all the time, being surrounded by coral reef
and all manner of calcified or silica-ized organisms. The problem is that
these tissues are not supposed to be mineralized, so I was wondering if it
is a chemical reaction that's making them hard.

A few responders wrote about similar experiences, citing various possible
reasons, but the one that resonates most is from Andrea Brothers, who
reminds me that mature coral nematocysts are nearly impenetrable to
solvents and epoxies. I suspect this red filamentous alga has hard fibrous
cell walls that are also impenetrable. This explanation fits both
situations. I am going to take Phil Oshel's advice and toss some of the
red algae into the SEM and do EDS to make sure it's not mineralized. The
clients insist it's not supposed to be.

Mahalo,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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From: mike.bode-at-resaltatech.com
Date: Thu, 14 Oct 2010 14:43:12 -0500
Subject: [Microscopy] viaWWW: Image analysis software

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

Hello Hany,

The OSIS Scandium software is specifically designed for SEM and with the
"solution Detection" has nearest neighbor measurements and statistics
directly included. You just set a threshold, select "Nearest Neighbor
distance" as one of the measurement criteria, and press a button to start
the analysis. You can find out more here:

http://www.resaltatech.com/scandium_sol_detection_main.htm

mike
---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com






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Name: Hany

Organization: University of Calgary

Title-Subject: [Filtered] Image analysis software

Message: Hi there,
I wonder what software you recommend for particle analysis on SEM
images. I am interested in getting particle size and spacing
distribution.
It does not have to be a free software, I can pay up to 500$ for a
student edition version.

Thanks a lot

Hany

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From: Colin.Veitch-at-csiro.au
Date: Thu, 14 Oct 2010 19:12:37 -0500
Subject: [Microscopy] Camera Mount For Olympus BH-2 microscope

Contents Retrieved from Microscopy Listserver Archives
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Hi,

A colleague has an Olympus BH-2 microscope and is looking for a camera mount. The part number from Olympus is U-PMTVC. Olympus has told him that they no longer have these in stock. Is there anyone out there who may have one that is no longer required or knows where on is that we can purchase? Alternatively, could someone suggest an alternative mount and source?

Thank you.

Colin Veitch

Electron Microscopist
CSIRO Materials Science and Engineering, Geelong Laboratory
PO Box 21, BELMONT, Vic. 3216. Australia.
colin.veitch-at-csiro.au
http://www.tft.csiro.au

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The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Materials Science and Engineering on +61 3 5246 4000.



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From: gary-at-gaugler.com
Date: Thu, 14 Oct 2010 19:19:48 -0500
Subject: [Microscopy] Re: Camera Mount For Olympus BH-2 microscope

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

I have one that is not being used. Like new.
I also have focusing telescopes for PM-10AD and ADS

gary g.


At 05:14 PM 10/14/2010, you wrote:



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From: jmircheski-at-us.es
Date: Fri, 15 Oct 2010 04:35:04 -0500
Subject: [Microscopy] viaWWW: gluteraldehyde

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

Hi Susan,

The issue of Glutaraldehyde aliquots at different concentrations, with or
without buffers is stability and efficiency of the fixative. With time, even
when stored at low temperatures, glutaraldehyde tends to polymerize. These
polymers decrease the fixative solution effectiveness. The higher the
concentration, the more polymerization you will have. It also depends on pH,
temperature and age of reagent. Concentrated glutaraldehyde at room
temperature polymerizes very fast, also when there are bases, acids or
oxygen present. If the solution is in low concentration, this is slowed
down. These solutions contain mostly the monomeric (active) form of the
fixative and are stable at pH 3-8 (as much as I can remember). If there is
water in the solution, glutaraldehyde tends to polymerize. On the other
hand, keeping working concentration of glutaraldehyde in buffers suffers
changes in the solution osmolarity. With time, the glutaraldehyde solutions
in buffers tend to increase their osmolarity, so you won't have equal fixing
conditions with aliquots from the same lot in different time intervals.
In general, if the colour of the glutaraldehyde is getting yellowish, don't
use it. In addition, if the pH is below 3, it's not good. What I usually do
is buy small volume aliquots of unbuffered 25% glutaraldehyde (usually in
ampoules), store them frozen and always prepare fresh fixative on the day of
the experiment.
Some time ago I got my hands on our "Bible" by Hayat, the fourth edition
from 2000. I too, needed information on the fixatives and I memorized these
things about the glutaraldehyde concentration and polymerization. If you
need further information, I recommend you to revert to this book. If anybody
else could recommend any other reading on this subject, please do that. I'm
always very happy to get new insights in this subject.

Regards,


Dr. Josif Mircheski
____________________________________________________________________________
___
Instituto de Biomedicina de Sevilla (IBIS)
Hosp.Univ. Virgen del Rocío/CSIC/Univ. de Sevilla y
Dpto. Fisiologia Médica y Biofísica
Universidad de Sevilla
Facultad de Medicina
Avda. Sánchez-Pizjuán 4
41009-Sevilla
 
Phone:+34-954556103
Fax:+34-954551769
e-mail: jmircheski-at-us.es

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Email: susan.trant-at-viha.ca
Name: Sue Trant

Organization: Vancouver Island Health Authority

Title-Subject: [Filtered] gluteraldehyde

Message: Hello all

We currently receive our gluteraldehyde in 8% - 10ml vials. We then
buffer them down and aliquot to a 3.5 % solution for use in the EM
lab. In hopes of maximizing our resources/time we are looking into
purchasing gluteraldehyde already aliquoted and diluted. I have seen
that there are vendors that supply 2.5 % gluteraldehyde in buffers.
What is the opinion of the lower concentration and does anyone
purchase these products?


Login Host: 207.194.133.9
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From: PhillipsT-at-missouri.edu
Date: Fri, 15 Oct 2010 09:06:51 -0500
Subject: [Microscopy] FW: glutaraldehyde

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

In regards to the stability of 8% glutaraldehyde -

Sigma Aldrich product data sheet states: "Purified samples of 8% glutaraldehyde stored at -20 C showed virtually no change in their UV absorbance characteristics even after 8 months" with this reference - Gillett, R., and Gull, K., Glutaraldehyde--Its Purity and Stability. Histochemie, 30, 162-167 (1972)

See also Don Ranly's informative article on the stability of glutaraladehyde: http://www.aapd.org/upload/articles/Ranly-06-02.pdf

I agree with Stephane that the amount of work required to dilute 8% glutaraldehyde is worth the effort. If you don't want to measure it out, design your protocol so that you use the entire ampoule at once.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/



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From: ph2-at-sprynet.com
Date: Fri, 15 Oct 2010 09:32:59 -0500
Subject: [Microscopy] FW: glutaraldehyde

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

A note regarding Glut degradation:

a. In a carefully monitored study, a 25% aqueous solution of
glutaraldehyde was purified to a single peak with a UV absorbance maximum of
280 nm (3). The subsequent detection of a second peak at 235 nm indicated
the formation of alternate forms, such as polymers, in investigations of the
influence of pH, temperature, and buffering on polymerization rate. No
polymerization occurred when a solution of glutaraldehyde was stored for 5
months at -14°C. There was a slight increase in the with storage at 4°C, and
then a rapid increase in this peak beginning with storage around 20°C
continuing to 60°C. The polymerization rate of glutaraldehyde was increased
when the pH was slightly acidic or basic; the rate polymerization was
decreased somewhat by the addition of buffers. If a 50% degree of
polymerization can be tolerated, samples may be stored at and pH 6.5 for up
to 7 months.

(3) Rasmussen, K.-E. and Albrcchtscn, J., Glutaraldehyde. The influence of
pH, temperature, and buffering on the polymerization rate. Histochemistry,
38, 19, 1974.


B. My experience in the tissue preservation for medical purposes at 4C
at {1% Glut buffered to 7.5-8 pH (sodium bicarb) will begin degrading in a
couple of weeks and continue until not useful at 3 months.



Tony


-----Original Message-----
X-from: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
Sent: Friday, October 15, 2010 10:12 AM
To: ph2-at-sprynet.com

In regards to the stability of 8% glutaraldehyde -

Sigma Aldrich product data sheet states: "Purified samples of 8%
glutaraldehyde stored at -20 C showed virtually no change in their UV
absorbance characteristics even after 8 months" with this reference -
Gillett, R., and Gull, K., Glutaraldehyde--Its Purity and Stability.
Histochemie, 30, 162-167 (1972)

See also Don Ranly's informative article on the stability of
glutaraladehyde: http://www.aapd.org/upload/articles/Ranly-06-02.pdf

I agree with Stephane that the amount of work required to dilute 8%
glutaraldehyde is worth the effort. If you don't want to measure it out,
design your protocol so that you use the entire ampoule at once.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/



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From: herro001-at-umn.edu
Date: Fri, 15 Oct 2010 14:11:55 -0500
Subject: [Microscopy] StrepTag II IFA ?

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

I am expressing a protein-StrepTag II fusion in a mammalian cell line
and I have been trying to visualize the localization using the IFA Kit
from IBA.
I have had NO luck with this and there is no positive control. Has
anyone else had any luck visualizing this epitope tag?


Michael J. Herron, U of MN, Dept. of Entomology
herro001-at-umn.edu
612-624-3688 (office) 612-625-5299 (FAX)




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From: William.F.Tivol-at-aero.org
Date: Fri, 15 Oct 2010 15:23:40 -0500
Subject: [Microscopy] New contact info

Contents Retrieved from Microscopy Listserver Archives
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Dear List,
I have just started working for Aerospace Corp. My phone number
is (310) 336-1991, and my smail address is 2310 E. El Segundo Blvd., Mail
Stop M2-244, El Segundo CA 90245.
Yours,
Bill

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From: kenp-at-labx.com
Date: Fri, 15 Oct 2010 16:36:37 -0500
Subject: [Microscopy] Camera Mount For Olympus BH-2 microscope

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

Not sure if this is the exact part you need, but you can try Spach Optics
who have this listed for sale on LabX... contact them using the form at
the bottom.
http://www.labx.com/v2/spiderdealer2/vistaSearchDetails.cfm?LVid=7894183

Alternately you can post a free wanted ad on LabX too if you need more
exposure for your request.

Best,
Ken Piech

General Manager
LabX Media Group
www.labx.com
www.labmanager.com
888-781-0328

-----Original Message-----
X-from: Colin.Veitch-at-csiro.au [mailto:Colin.Veitch-at-csiro.au]
Sent: October-14-10 8:24 PM
To: kenp-at-labx.com

Hi,

A colleague has an Olympus BH-2 microscope and is looking for a camera
mount. The part number from Olympus is U-PMTVC. Olympus has told him
that they no longer have these in stock. Is there anyone out there who
may have one that is no longer required or knows where on is that we can
purchase? Alternatively, could someone suggest an alternative mount and
source?

Thank you.

Colin Veitch

Electron Microscopist
CSIRO Materials Science and Engineering, Geelong Laboratory PO Box 21,
BELMONT, Vic. 3216. Australia.
colin.veitch-at-csiro.au
http://www.tft.csiro.au

Tel:       +61 (0) 3 5246 4000
Mobile:  0438 538 475
Fax:      +61 (0) 3 5246 4811

The information contained in this e-mail message may be privileged or
confidential information. If you are not an intended recipient, you may
not copy, distribute or take any action in reliance on it. If you have
received this message in error, please telephone CSIRO Materials Science
and Engineering on +61 3 5246 4000.



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From: kenp-at-labx.com
Date: Fri, 15 Oct 2010 17:06:15 -0500
Subject: [Microscopy] Camera Mount For Olympus BH-2 microscope

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

Not sure if this is the exact part you need, but you can try Spach Optics
who have this listed for sale on LabX... contact them using the form at
the bottom.
http://www.labx.com/v2/spiderdealer2/vistaSearchDetails.cfm?LVid=7894183

Alternately you can post a free wanted ad on LabX too if you need more
exposure for your request.

Best,
Ken Piech

General Manager
LabX Media Group
www.labx.com
www.labmanager.com
888-781-0328

-----Original Message-----
X-from: Colin.Veitch-at-csiro.au [mailto:Colin.Veitch-at-csiro.au]
Sent: October-14-10 8:24 PM
To: kenp-at-labx.com

Hi,

A colleague has an Olympus BH-2 microscope and is looking for a camera
mount. The part number from Olympus is U-PMTVC. Olympus has told him
that they no longer have these in stock. Is there anyone out there who
may have one that is no longer required or knows where on is that we can
purchase? Alternatively, could someone suggest an alternative mount and
source?

Thank you.

Colin Veitch

Electron Microscopist
CSIRO Materials Science and Engineering, Geelong Laboratory PO Box 21,
BELMONT, Vic. 3216. Australia.
colin.veitch-at-csiro.au
http://www.tft.csiro.au

Tel:       +61 (0) 3 5246 4000
Mobile:  0438 538 475
Fax:      +61 (0) 3 5246 4811

The information contained in this e-mail message may be privileged or
confidential information. If you are not an intended recipient, you may
not copy, distribute or take any action in reliance on it. If you have
received this message in error, please telephone CSIRO Materials Science
and Engineering on +61 3 5246 4000.



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From: speransv-at-mail.nih.gov
Date: Fri, 15 Oct 2010 18:56:39 -0500
Subject: [Microscopy] FW: glutaraldehyde

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

Sorry if I did not read carefully, but I did not notice anyone pointing out yet that in the sealed ampules, unbuffered GA is kept under inert gas. Therefore, it should last longer.
Otherwise, my understanding is that lower concentrations of such unbuffered stock last longer than more concentrated ones, because polymerization rate is slower.

There is a good discussion, with literature references, on the subject of GA fixation in Gareth Griffiths' Fine Structure Immunocytochemistry book.

________________________________________________
Vlad Speransky, Staff Scientist
Biomedical Engineering and Physical Science Shared Resource
National Institute of Biomedical Imaging and Bioengineering, NIH
13 South Dr, Rm. 3N17 MSC 5766
Bethesda, MD 20892
301 496-3989
vladislav_speransky-at-nih.gov
http://www.nibib.nih.gov/Research/Intramural/SharedResource/Speransky

Opinions and experiences related are those of Vlad Speransky and do not represent the NIH. On the good side, this message is not confidential and can be freely shared and reproduced.


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A note regarding Glut degradation:

a. In a carefully monitored study, a 25% aqueous solution of
glutaraldehyde was purified to a single peak with a UV absorbance maximum of
280 nm (3). The subsequent detection of a second peak at 235 nm indicated
the formation of alternate forms, such as polymers, in investigations of the
influence of pH, temperature, and buffering on polymerization rate. No
polymerization occurred when a solution of glutaraldehyde was stored for 5
months at -14°C. There was a slight increase in the with storage at 4°C, and
then a rapid increase in this peak beginning with storage around 20°C
continuing to 60°C. The polymerization rate of glutaraldehyde was increased
when the pH was slightly acidic or basic; the rate polymerization was
decreased somewhat by the addition of buffers. If a 50% degree of
polymerization can be tolerated, samples may be stored at and pH 6.5 for up
to 7 months.

(3) Rasmussen, K.-E. and Albrcchtscn, J., Glutaraldehyde. The influence of
pH, temperature, and buffering on the polymerization rate. Histochemistry,
38, 19, 1974.


B. My experience in the tissue preservation for medical purposes at 4C
at {1% Glut buffered to 7.5-8 pH (sodium bicarb) will begin degrading in a
couple of weeks and continue until not useful at 3 months.



Tony


-----Original Message-----
X-from: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
Sent: Friday, October 15, 2010 10:12 AM
To: ph2-at-sprynet.com

In regards to the stability of 8% glutaraldehyde -

Sigma Aldrich product data sheet states: "Purified samples of 8%
glutaraldehyde stored at -20 C showed virtually no change in their UV
absorbance characteristics even after 8 months" with this reference -
Gillett, R., and Gull, K., Glutaraldehyde--Its Purity and Stability.
Histochemie, 30, 162-167 (1972)

See also Don Ranly's informative article on the stability of
glutaraladehyde: http://www.aapd.org/upload/articles/Ranly-06-02.pdf

I agree with Stephane that the amount of work required to dilute 8%
glutaraldehyde is worth the effort. If you don't want to measure it out,
design your protocol so that you use the entire ampoule at once.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/



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From: pmcooke-at-earthlink.net
Date: Fri, 15 Oct 2010 20:10:12 -0500
Subject: [Microscopy] Camera Mount For Olympus BH-2 microscope

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


Based on the specific information you have provided, as I understand it:
wanting Olympus a U-PMTVC adaptor for an Olympus BH-2 series microscope -
you want a C-mount adaptor. I assume your friend has a camera that is
C-mount and the Olympus trinoc-head will accept this adaptor (Olympus
offered 2 models of trinoc-heads for the BH-2 series and the two accept
different type adaptors.) These adaptors show up on Ebay all the time. They
go by "MTV-3 adaptor." See the link to Ebay - and this is about what they go
for on Ebay (unless you're lucky or your timing is just right):

http://cgi.ebay.com/ws/eBayISAPI.dll?ViewItem&Item=300481162659&Category=118
13&_trkparms=algo%3DLVI%26its%3DI%26otn%3D1#ht_500wt_1130

You will also need a photo-eyepiece like this, which show up on Ebay all the
time, and, yes, at about this price-point:

http://cgi.ebay.com/Olympus-PE-3-3X-Photo-Eyepiece-/130422433691?pt=LH_Defau
ltDomain_0&hash=item1e5dc8679b#ht_500wt_1130

The photo-eyepieces come in various magnifications, 1.67, 2.5, 3.3, 5.0, 7.6
so you'll want to know what magnification you want for projection (and
higher mag is not always better as it totally depends on your projected
intentions).

Hope this helps,

Peter

Peter M. Cooke
MICA

2) The thread:



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Not sure if this is the exact part you need, but you can try Spach Optics
who have this listed for sale on LabX... contact them using the form at
the bottom.
http://www.labx.com/v2/spiderdealer2/vistaSearchDetails.cfm?LVid=7894183

Alternately you can post a free wanted ad on LabX too if you need more
exposure for your request.

Best,
Ken Piech

General Manager
LabX Media Group
www.labx.com
www.labmanager.com
888-781-0328

-----Original Message-----
X-from: Colin.Veitch-at-csiro.au [mailto:Colin.Veitch-at-csiro.au]
Sent: October-14-10 8:24 PM
To: kenp-at-labx.com

Hi,

A colleague has an Olympus BH-2 microscope and is looking for a camera
mount. The part number from Olympus is U-PMTVC. Olympus has told him
that they no longer have these in stock. Is there anyone out there who
may have one that is no longer required or knows where on is that we can
purchase? Alternatively, could someone suggest an alternative mount and
source?

Thank you.

Colin Veitch

Electron Microscopist
CSIRO Materials Science and Engineering, Geelong Laboratory PO Box 21,
BELMONT, Vic. 3216. Australia.
colin.veitch-at-csiro.au
http://www.tft.csiro.au

Tel: +61 (0) 3 5246 4000
Mobile: 0438 538 475
Fax: +61 (0) 3 5246 4811

The information contained in this e-mail message may be privileged or
confidential information. If you are not an intended recipient, you may
not copy, distribute or take any action in reliance on it. If you have
received this message in error, please telephone CSIRO Materials Science
and Engineering on +61 3 5246 4000.



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From: FMonson-at-wcupa.edu
Date: Fri, 15 Oct 2010 20:31:24 -0500
Subject: [Microscopy] FW: glutaraldehyde

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

In my experience, sealed ampules of glut under nitrogen will last as long as the ampule itself does not leak. At least, I have not seen any difference that I can detect in the results.

Cheers,

Fred Monson
________________________________________
X-from: PhillipsT-at-missouri.edu [PhillipsT-at-missouri.edu]
Sent: Friday, October 15, 2010 10:16 AM
To: Monson, Frederick

In regards to the stability of 8% glutaraldehyde -

Sigma Aldrich product data sheet states: "Purified samples of 8% glutaraldehyde stored at -20 C showed virtually no change in their UV absorbance characteristics even after 8 months" with this reference - Gillett, R., and Gull, K., Glutaraldehyde--Its Purity and Stability. Histochemie, 30, 162-167 (1972)

See also Don Ranly's informative article on the stability of glutaraladehyde: http://www.aapd.org/upload/articles/Ranly-06-02.pdf

I agree with Stephane that the amount of work required to dilute 8% glutaraldehyde is worth the effort. If you don't want to measure it out, design your protocol so that you use the entire ampoule at once.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/



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7, 31 -- From: "Phillips, Thomas E." {PhillipsT-at-missouri.edu}
7, 31 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com} ,
7, 31 -- "susan.trant-at-viha.ca" {susan.trant-at-viha.ca}
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14, 27 -- From FMonson-at-wcupa.edu Fri Oct 15 20:31:24 2010
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14, 27 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
14, 27 -- Date: Fri, 15 Oct 2010 21:31:21 -0400
14, 27 -- Subject: RE: [Microscopy] FW: glutaraldehyde
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From: schooley-at-mcn.org
Date: Sun, 17 Oct 2010 12:39:48 -0500
Subject: [Microscopy] Education

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

Project MICRO (Microscopy In Curriculum -
Research Outreach) is an MSA educational outreach
program. MICRO's website (URL below) has a lot
of information, including an extensive reviewed
booklist. In the last few years, K-12
nanotechnology education books have started to
appear; they're closely related to microscopy,
but different enough to deserve their own list.
MICRO has that too! 'Nanotechnology for Kids' is
available on the MICRO site, and 6 more book
titles have just been added to the original dozen.

While you're visiting the MICRO site, look up
'Unseen Companions' in the main booklist; it's
the ultimate SEM coffee table book. Beautiful,
if you like ugly bugsŠ

MICRO began by sponsoring an outstanding
teachers' guide, 'Microscopic Explorations', it
was published in '98. It's been very successful,
but detailed science curricula have fallen on
hard times; in too many schools 'test prep'
consumes the time that once was available for
science education. There is a very effective,
independently published guide, 'Private Eye',
that is easier to use than 'Microscopic
Explorations' in many situations, and two YouTube
descriptions have just appeared; if you're doing
or planning any outreach you'll find them very
interesting:

The Private Eye and its Training-the-Trainers
Institute: http://www.youtube.com/watch?v=G
0qC44Og0Q

The Private Eye and afterschool programs:
http://www.sedl.org/cgi-bin/mysql/afterschool/
science.cgi?location=search&show_resource_id=28

And a final item: The Royal Microscopical Society
has an outreach program that is as old as MICRO.
It was called "A Microscope for Every School" in
the past, but program emphasis has changed
recently to an emphasis on working with schools.
The current issue of the RMS members' journal,
"infocus", has an article on outreach titled
"Stepping inside the cell"; you can read it at
http://www.rms.org.uk/publications/infocus

--
Caroline Schooley
Project MICRO Coordinator
Microscopy Society of America
45301 Caspar Point Road, Box 117
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.microscopy.org/ProjectMICRO


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From: nizets2-at-yahoo.com
Date: Mon, 18 Oct 2010 05:36:20 -0500
Subject: [Microscopy] FW: glutaraldehyde

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

If the polymerization of glutaraldehyde is not an oxidization process, there is
no reason why sealing in inert gas should change something.
Anyway I have been told in my learning age that highly concentrated glutar was
more stable than diluted one and so I am interested in this discussion.
Fact is, it must be diluted in water and not in buffer if I understood well.

Stephane



----- Original Message ----
X-from: "FMonson-at-wcupa.edu" {FMonson-at-wcupa.edu}
To: nizets2-at-yahoo.com
Sent: Sat, October 16, 2010 3:34:39 AM

In my experience, sealed ampules of glut under nitrogen will last as long as the
ampule itself does not leak.  At least, I have not seen any difference that I
can detect in the results.

Cheers,

Fred Monson
________________________________________
X-from: PhillipsT-at-missouri.edu [PhillipsT-at-missouri.edu]
Sent: Friday, October 15, 2010 10:16 AM
To: Monson, Frederick

In regards to the stability of 8% glutaraldehyde -

Sigma Aldrich product data sheet states: "Purified samples of 8% glutaraldehyde
stored at -20 C showed virtually no change in their UV absorbance
characteristics even after 8 months" with this reference - Gillett, R., and
Gull, K., Glutaraldehyde--Its Purity and Stability. Histochemie, 30, 162-167
(1972)

See also Don Ranly's informative article on the stability of glutaraladehyde:
http://www.aapd.org/upload/articles/Ranly-06-02.pdf

I agree with Stephane that the amount of work required to dilute 8%
glutaraldehyde is worth the effort. If you don't want to measure it out, design
your protocol so that you use the entire ampoule at once.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/



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7, 31 -- From: "Phillips, Thomas E." {PhillipsT-at-missouri.edu}
7, 31 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com} ,
7, 31 --        "susan.trant-at-viha.ca" {susan.trant-at-viha.ca}
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14, 27 -- From: "Monson, Frederick" {FMonson-at-wcupa.edu}
14, 27 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
14, 27 -- Date: Fri, 15 Oct 2010 21:31:21 -0400
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28, 35 -- From: Stephane Nizet {nizets2-at-yahoo.com}
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From: PhillipsT-at-missouri.edu
Date: Mon, 18 Oct 2010 08:12:18 -0500
Subject: [Microscopy] FW: glutaraldehyde

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

I could be wrong but I always assumed that there were two problems
with glutaraldehyde "going off".
One is the polymerization problem and the other oxidation to acid -
hence the nitrogen atmosphere to limit problem two but not one.

It's a while since I read the Hayat and Glauert books so my memory
could be deceiving me.

Malcolm

Malcolm Haswell
Imaging Suite
Faculty of Applied Sciences
University of Sunderland
SUNDERLAND
SR1 3SD
UK


----- Original Message -----
X-from: nizets2-at-yahoo.com

I am not sure what you mean by "Fact is, it must be diluted in water and not in buffer..." - Clearly I dilute my glutaraldehyde into the buffer I use.

Unlike you, I have a strong recollection of reading that 8% glutaraldehyde was more stable than more concentrated versions but can't find any reference for this statement.



Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Monday, October 18, 2010 5:38 AM
To: Phillips, Thomas E.

If the polymerization of glutaraldehyde is not an oxidization process, there is
no reason why sealing in inert gas should change something.
Anyway I have been told in my learning age that highly concentrated glutar was
more stable than diluted one and so I am interested in this discussion.
Fact is, it must be diluted in water and not in buffer if I understood well.

Stephane



----- Original Message ----
X-from: "FMonson-at-wcupa.edu" {FMonson-at-wcupa.edu}
To: nizets2-at-yahoo.com
Sent: Sat, October 16, 2010 3:34:39 AM

In my experience, sealed ampules of glut under nitrogen will last as long as the
ampule itself does not leak.  At least, I have not seen any difference that I
can detect in the results.

Cheers,

Fred Monson
________________________________________
X-from: PhillipsT-at-missouri.edu [PhillipsT-at-missouri.edu]
Sent: Friday, October 15, 2010 10:16 AM
To: Monson, Frederick

In regards to the stability of 8% glutaraldehyde -

Sigma Aldrich product data sheet states: "Purified samples of 8% glutaraldehyde
stored at -20 C showed virtually no change in their UV absorbance
characteristics even after 8 months" with this reference - Gillett, R., and
Gull, K., Glutaraldehyde--Its Purity and Stability. Histochemie, 30, 162-167
(1972)

See also Don Ranly's informative article on the stability of glutaraladehyde:
http://www.aapd.org/upload/articles/Ranly-06-02.pdf

I agree with Stephane that the amount of work required to dilute 8%
glutaraldehyde is worth the effort. If you don't want to measure it out, design
your protocol so that you use the entire ampoule at once.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/



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From: beth-at-plantbio.uga.edu
Date: Mon, 18 Oct 2010 09:45:35 -0500
Subject: [Microscopy] acetonitrile?

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

Hi all,
Does anyone use acetonitrile for processing samples for TEM?
A friend wants to know if they can use acetonitrile instead of
propylene oxide. I've never used it before though I bet is is okay.
The resin they plan to use is Embed 812.
thanks for any advice,
Beth

} have you ever used acetonitrile? supposed to be a replacement for
} PO.....


==============================Original Headers==============================
3, 19 -- From beth-at-plantbio.uga.edu Mon Oct 18 09:45:34 2010
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From: TindallR-at-missouri.edu
Date: Mon, 18 Oct 2010 09:54:56 -0500
Subject: [Microscopy] acetonitrile?

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

Hi Beth,

If the goal here is to avoid the use of PO, you can simply do all the processing steps in acetone, rather than ethanol. I have also heard that acetonitrile was a PO substitute, but I don't really have any experience along these lines. We haven't used PO since, like, forever.

Also, I have several times transitioned from 100% ethanol into epoxy resins, generally Epon/Araldite or Epon/Spurrs, with no problems. (I can almost hear the gasps of horror from here......!) If you want to try this, I would recommend a couple extra infiltration steps with pure resin before polymerization, and try it with a test sample before risking your Nobel Prize one-of-a-kind specimens in it.

Good luck and

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




-----Original Message-----
X-from: beth-at-plantbio.uga.edu [mailto:beth-at-plantbio.uga.edu]
Sent: Monday, October 18, 2010 9:47 AM
To: Tindall, Randy D.

Hi all,
Does anyone use acetonitrile for processing samples for TEM?
A friend wants to know if they can use acetonitrile instead of
propylene oxide. I've never used it before though I bet is is okay.
The resin they plan to use is Embed 812.
thanks for any advice,
Beth

} have you ever used acetonitrile? supposed to be a replacement for
} PO.....


==============================Original Headers==============================
3, 19 -- From beth-at-plantbio.uga.edu Mon Oct 18 09:45:34 2010
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From: reinhard.rachel-at-biologie.uni-regensburg.de
Date: Mon, 18 Oct 2010 09:56:58 -0500
Subject: [Microscopy] Antw: acetonitrile?

Contents Retrieved from Microscopy Listserver Archives
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} } }
} Does anyone use acetonitrile for processing samples for TEM?
} A friend wants to know if they can use acetonitrile instead of
} propylene oxide. I've never used it before though I bet is is okay.
} The resin they plan to use is Embed 812.
} thanks for any advice,

I think it is commonly known now that the propylene oxide step can safely be replaced by one (or two) washing step(s) in pure acetone, followed by stepwise infiltration of Epon resin (similar to / identical with (?) Embed 812). At least, we do this since many years now, and similarly, many others, with success.

kind regards,
Reinhard


--
PD Dr. Reinhard Rachel
Universitaet Regensburg
Centre for EM / Anatomy
Faculty for Biology&Preclin. Med.
Universitaetsstr. 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29



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From: nizets2-at-yahoo.com
Date: Mon, 18 Oct 2010 09:57:08 -0500
Subject: [Microscopy] acetonitrile?

Contents Retrieved from Microscopy Listserver Archives
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Following a suggestion made on this list, I made a try by replacing PO with
acetonitrile.
The blocks did not polymerize well. I only tried once, though. But usually I
never have polymerization problems.

regards,
Stephane



----- Original Message ----
X-from: "beth-at-plantbio.uga.edu" {beth-at-plantbio.uga.edu}
To: nizets2-at-yahoo.com
Sent: Mon, October 18, 2010 4:50:10 PM

Hi all,
Does anyone use acetonitrile for processing samples for TEM?
A friend wants to know if they can use acetonitrile instead of 
propylene oxide. I've never used it before though I bet is is okay. 
The resin they plan to use is Embed 812.
thanks for any advice,
Beth

} have you ever used acetonitrile?  supposed to be a replacement for 
} PO.....


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From: nizets2-at-yahoo.com
Date: Mon, 18 Oct 2010 10:08:11 -0500
Subject: [Microscopy] cell density using LM

Contents Retrieved from Microscopy Listserver Archives
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Hello dear listers!

I need to quantify the density of my cells in culture but they are hard to count
on the classical Thomas cell counter because they stay in clusters.
I wondered if I could "simply" use our light microscope to do it.
We have a Zeiss axiovert 200M with a nice software, perhaps the software could
count the cells automatically?
Has anyone experience using a LM to quantify the cell density (automatically
would be better)?
I would be grateful if you could share your thoughts (even if you tell me that
this idea is crazy).

best regards,
Stephane




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From: W.Muss-at-salk.at
Date: Mon, 18 Oct 2010 10:34:39 -0500
Subject: [Microscopy] Re: acetonitrile?

Contents Retrieved from Microscopy Listserver Archives
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Hi, all....

Dear Beth,
thios matter has been discussed a while ago on the list server.
I use Acetonitrile (AN) instead of PO since 1988 or 1989 without any problems (and have posted shorter and longer messages on the list server too). Some rules different from PO have to be followed, one of them would be:
no dehydration by acetonitrile like dehydration with ascending ethanols or acetone...instead using AN only as an intermediate after EtOH 100% or Acetone 100%.

EtOH 100% (2-3 times -at- RT)
AN I (5-10 min -at- RT)
AN II (10-15 min -at- RT)
AN III (10-15 min -at-RT)
times / step depending on size of specimens. Use probe-rotator (and use glass vials with carefully closed lid/cap all the time).
Cautions for personal protection (AN = methylcyanide) are to be taken: use only in a ventilated area (e.g. in or just in front of a fume-cupboard).
Remember also:
AN is 100% water-miscible (cave relative humidity!).
After step AN III pour out AN pure and imbibe specimens in the vials with
AN(pure) - pure resin = 1:1, perhaps use/work under a "lamp" to raise the temperature above specs a little bit, especially when you open lids of vials after a 30-45 min infiltration).
Then, under the lamp you can rotate the open glass vials for - say - 30 mins,
then transfer specs into separate infiltration forms/cups filled with Resin pure I, II, III (each step ca. 45min to 1 h)... You can help the residual AN evaporate by placing the specs in pure resin in a polymerizing oven at e.g. 37 degr. C (as before: 45 - 60 min each step, every step use a new "infiltrating mold or form/cup) usually present in an EM-Lab.

I use the Epon 812 substitute "Glycidether 100" from SEVA, Heidelberg, Germany (Glycidether 100, DDSA, MNA and DMP-30, classical formulation) and do have, I think good results.
If interested in further, more specific info, please let me know, I should be glad to help.

Best regards,

Wolfgang MUSS PhD
EM-Lab, Pathology
SALK-LKH (Gen.Hosp) & PMU
SALZBURG, Austria

PS: since I have finished a photo-session on diseased nervus suralis, I should be glad to send you (by a separate mail) a semithin image of that nerve, just to show you what the "quality" of "big specimens" is....



} -----Ursprüngliche Nachricht-----
} Von: beth-at-plantbio.uga.edu [mailto:beth-at-plantbio.uga.edu]
} Gesendet: Montag, 18. Oktober 2010 16:49
} An: Muß Wolfgang
} Betreff: [Microscopy] acetonitrile?
}
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} Hi all,
} Does anyone use acetonitrile for processing samples for TEM?
} A friend wants to know if they can use acetonitrile instead of
} propylene oxide. I've never used it before though I bet it is okay.
} The resin they plan to use is Embed 812.
} thanks for any advice,
} Beth
}
} } have you ever used acetonitrile? supposed to be a replacement for
} } PO.....
}
}
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} Headers==============================
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From: beth-at-plantbio.uga.edu
Date: Mon, 18 Oct 2010 10:53:20 -0500
Subject: [Microscopy] thanks for acetonitrile responses

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

Thanks to y'all for taking the time to respond to my question. They
have been forwarded to the person in need.
If this subject was recently discussed on the list then I apologize
for not paying attention to the posts at that time.

my best,
Beth








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From: cljohnson33-at-gmail.com
Date: Mon, 18 Oct 2010 10:55:26 -0500
Subject: [Microscopy] Drexel Univ & Philadelphia Society for Microscopy Symposium

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

Greetings Listers,

You are cordially invited to attend the “Applications of Microscopy in
Materials and Biological Sciences” symposium on Monday, November 8,
2010 at Drexel University. The symposium is hosted jointly by Drexel’s
Centralized Research Facility and the Philadelphia Society for
Microscopy. Please go to http://crf.coe.drexel.edu/crf/workshops to
register for free by October 24. Space is limited. For more
information please contact Craig Johnson at (215) 895-5900 or
cljohnson-at-coe.drexel.edu.


Applications of Microscopy in Materials and Biological Sciences

Monday, November 8, 2010
Drexel University, Bossone Research Enterprise Center
3126 Market Street, Philadelphia, PA


8:30 am                Registration and Coffee

Morning Session

9:00 am                Opening Remarks – Dr. Deborah Crawford, Vice
Provost for Research, Drexel University and Robert Carlton, President
Philadelphia Society for Microscopy

9:10 am                Electrons, Camera, Action! Advanced Multi-scale
Microscopy Techniques for Understanding Microstructure Property
Relationships in Materials – Dr. Mitra Taheri, Drexel University

9:50 am                NanoMicro-Structured Silicon: Pits, Pillars,
Pores and Powder – Dr. Kurt Kolasinski, West Chester University

10:30 am              Coffee Break

11:10 am              Vibrational Microscopy in Forensics – Dr.
Pauline Leary, John Jay College/Smith’s Detection

11:50                    Lunch

Afternoon Session

1:00 pm                Keynote Presentation: Microanalysis of Martian
Salts from the Comfort of Your Laboratory – Dr. Edward Vincenzi,
Smithsonian Institution

2:00 pm                Real-World Strategies for Correlative
Microscopy – Dr. Kurt Czymmek, University of Delaware

2:40 pm                Coffee Break, Tours of the Centralized Research
Facilities

3:15 pm                Philadelphia Society for Microscopy Business Meeting

Sponsored by: Angstrom Scientific, Electron Microscopy Sciences, FEI,
Gatan, Hysitron, JEOL, Leica Microsystems, Oxford Instruments,
Protochips, Smiths Detection, and SPI


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From: biology-at-ucla.edu
Date: Mon, 18 Oct 2010 11:24:25 -0500
Subject: [Microscopy] Section compression ...

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

Hi All,

Lately I am having a issue with ultrathin section compression. The
sections are not as flat as I would like when picked up on the grid. I
know that using chloroform vapors will flatten out the sections.

Problem I am having is locating a specific type or vial and cap that i
had used in the pat to keep the chloroform. I used to have this glass
vial with what appeared to be a cork like cap and attached to the cap
was either a brush or applicator that was soaking in the chloroform.
When I removed the cap the applicator was attached and wet with
chloroform. This was just waved over the boat and the vapors from the
chloroform flattened out the sections.

It was reusable instead of using a cotton applicator every time.. I am
looking for this type or vial but, I have no idea where to look for it.

I have tried Electron Microscopy Sicence , Ted Pella already. Any
help would be appreciated..

Thanks
Eric A. Rosen
Electron Microscopy Lab
Dept. Pathology
UCLA medical Center

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From: FMonson-at-wcupa.edu
Date: Mon, 18 Oct 2010 12:51:31 -0500
Subject: [Microscopy] FW: glutaraldehyde

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

While there are several to many explanations for initiations of polymerizations of various compositions, the requirements for HCHO are unique, because the native state (at STP) for HCHO is a gas. While, again, I have not been able to sort thru ALL of the literature, my memory is jogged sufficiently to remind me that most studies of polymeric reactions involving HCHO do not address the self-polymerization of the substance.

I have been in my present location for 10 years, and there are 3-4 packs of 10ml glass vials of 50% glutaraldehyde marked with the phrase, "sealed under nitrogen" or something to that effect (I am operating without perfect memory or immediate access to them. In the last 20 years, I have picked one and then another of these packages from colleagues who are moving or closing their labs. In all of that time, I have only experienced one loss of a vial to polymerization (quantitative since there was no odor of 'glut' when I cracked it to deactivate the substance).

To summarize what I now remember, I offer the following.

Even in 50% aqueous solutions of glutaraldehyde or HCHO, the key to stability appears to be oxygen. Further, the polymerization can be photo-catalyzed. Thus, in addition to a nitrogen environment, my vials are made of brown glass. So, I suggest that by removing oxygen via nitrogen purging and keeping the substance (8-50%) in the dark, one may expect that the common initiator of reactions - free radical oxygen species - will not be created in quantities sufficient to initiate noticeable polymerization OR oxidation. The other characteristic of aldehyde polymerizations is low pH. HCHO + O -} HCHOOH (formic acid).

I have not used Formalin for fixation of any kind since the mid 1960's, since the day I was asked to tap a 55 gallon container and received a trickle of liquid and lots of white flakes.

When I prepare HCHO from Paraformaldehyde, I prepare 200mls -at- 20% (by weight), and I store it for no more than 6months in 50-100ml screw cap jars with PTFE-lined caps - sealed for id with a wrap of 'Parafilm'.

Cheers, and I am not an organic chemist, but I did take "Organic Mechanisms" (4-500 course?) - achieved a 77 average and was 3rd in a class of 300. As a biologist, in his late 20's in the late 1960's, I was the only one who didn't complain, because I thought my grade was well earned and properly recorded.

Fred Monson (fmonson-at-wcupa.edu)
http://cmirt.wcupa.edu


________________________________________
X-from: PhillipsT-at-missouri.edu [PhillipsT-at-missouri.edu]
Sent: Monday, October 18, 2010 9:20 AM
To: Monson, Frederick

I am not sure what you mean by "Fact is, it must be diluted in water and not in buffer..." - Clearly I dilute my glutaraldehyde into the buffer I use.

Unlike you, I have a strong recollection of reading that 8% glutaraldehyde was more stable than more concentrated versions but can't find any reference for this statement.



Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Monday, October 18, 2010 5:38 AM
To: Phillips, Thomas E.

If the polymerization of glutaraldehyde is not an oxidization process, there is
no reason why sealing in inert gas should change something.
Anyway I have been told in my learning age that highly concentrated glutar was
more stable than diluted one and so I am interested in this discussion.
Fact is, it must be diluted in water and not in buffer if I understood well.

Stephane



----- Original Message ----
X-from: "FMonson-at-wcupa.edu" {FMonson-at-wcupa.edu}
To: nizets2-at-yahoo.com
Sent: Sat, October 16, 2010 3:34:39 AM

In my experience, sealed ampules of glut under nitrogen will last as long as the
ampule itself does not leak. At least, I have not seen any difference that I
can detect in the results.

Cheers,

Fred Monson
________________________________________
X-from: PhillipsT-at-missouri.edu [PhillipsT-at-missouri.edu]
Sent: Friday, October 15, 2010 10:16 AM
To: Monson, Frederick

In regards to the stability of 8% glutaraldehyde -

Sigma Aldrich product data sheet states: "Purified samples of 8% glutaraldehyde
stored at -20 C showed virtually no change in their UV absorbance
characteristics even after 8 months" with this reference - Gillett, R., and
Gull, K., Glutaraldehyde--Its Purity and Stability. Histochemie, 30, 162-167
(1972)

See also Don Ranly's informative article on the stability of glutaraladehyde:
http://www.aapd.org/upload/articles/Ranly-06-02.pdf

I agree with Stephane that the amount of work required to dilute 8%
glutaraldehyde is worth the effort. If you don't want to measure it out, design
your protocol so that you use the entire ampoule at once.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/



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From: erosen-at-mednet.ucla.edu
Date: Mon, 18 Oct 2010 18:48:08 -0500
Subject: [Microscopy] viaWWW: Section compression ...

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Email: erosen-at-mednet.ucla.edu
Name: Eric Rosen

Organization: UCLA Medical Center

Title-Subject: [Filtered] Section compression ...

Message: Hi All,

Lately I am having a issue with ultrathin section compression. The
sections are not as flat as I would like when picked up on the grid.

I know that using chloroform vapors will flatten out the sections.
Problem I am having is locating a specific type or vial and cap that
i had used in the pat to keep the chloroform.

I used to have this glass vial with what appeared to be a cork like
cap and attached to the cap was either a brush or applicator that was
soaking in the chloroform. When I removed the cap the applicator was
attached and wet with chloroform. This was just waved over the boat
and the vapors from the chloroform flattened out the sections. It was
reusable instead of using a cotton applicator every time..

I am looking for this type or vial but, I have no idea where to look
for it. I have tried Electron Microscopy Sicence , Ted Pella already.

Any help would be appreciated..

Thanks
Eric A. Rosen
Electron Microscopy Lab
Dept. Pathology
UCLA medical Center

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From: leech-at-sfsu.edu
Date: Mon, 18 Oct 2010 18:49:05 -0500
Subject: [Microscopy] viaWWW: Full-time Electron Microscopy Facility Manager position

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Email: leech-at-sfsu.edu
Name: Mary Leech

Organization: San Francisco State University

Title-Subject: [Filtered] Full-time Electron
Microscopy Facility Manager position in San
Francisco

Message: Full-time Electron Microscopy Facility
Manager position available at San Francisco State
University

Position Summary: This position supports teaching
and research efforts of the College of Science,
specifically the Departments of Chemistry &
Biochemistry, Biology, Geosciences, Physics, and
School of Engineering, by managing the Electron
Microscopy Facility, instructing students, staff,
and faculty in the use of the scanning electron
microscope (SEM) including EDS, EBSD, CL, and
STEM detectors, sample preparation equipment, and
developing and maintaining protocols for
efficient and safe use of the equipment and
chemicals related to electron microscopy.

Description: Full-time temporary position with
the possibility of becoming permanent with a
successful candidate.
Classification Salary Range: $4,417-$6,625 per month.
Anticipated Hiring Range: $4,417 - $4,969 per month

PLEASE NOTE: This position is open until filled
and applications will be reviewed starting Nov. 1.

Minimum qualifications: BS or BA in physics,
chemistry, materials science, geology, biology,
or closely related field. Work Experience: 2-3
years of experience in the maintenance and
operation of SEM (scanning electron microscope),
EDS (energy dispersive spectroscopy), EBSD
(electron back-scatter diffraction), CL
(cathodoluminescence), and/or STEM (scanning
transmission) detectors, and sample preparation
equipment.

Preferred qualifications: MS or PhD in a field of
physical science; facility management experience;
extensive experience with SEM (scanning electron
microscope) on a wide variety of sample types;
experience with EDS (energy dispersive
spectroscopy), EBSD (electron back-scatter
diffraction), cathodoluminescence (CL), and
scanning transmission (STEM) accessories; working
knowledge/use of computers; basic knowledge of
electronics; trouble-shooting and repair skills.

Full position description available at:
https://cmsweb.sfsu.edu/psp/HSFPRDF/CUSTOMER/HRMS/c/HRS_HRAM.HRS_CE.GBL,
Research Technician III ñ College of Science &
Engineering, Job ID: 2338

Interested individuals may contact Prof. Mary
Leech at leech-at-sfsu.edu for more information.

How To Apply:
Submit an application and/or resume and cover
letter (optional), describing your specific
qualifications for this position.

All applicants must submit a SF State
Staff/Administrator Application and/or resume
with an original signature for each job posting.
All application material(s) must include the job
posting number (Job ID: 2338). To be considered,
application material must be submitted in person
or through U.S. mail to Human Resources; San
Francisco State University does not accept
on-line, e-mail, or faxed application materials
at this time.

Mail application material(s) to:

San Francisco State University
Human Resources, Safety & Risk Management - Staff Employment Services
1600 Holloway Avenue, Administration Building 252,
San Francisco, CA 94132-4252

The Human Resources, Safety & Risk Management
office is open Mondays, Tuesdays, Thursdays and
Fridays from 8 a.m. to 5 p.m., and can be reached
at (415) 338-1872 or (415) 338-1873; TDD (415)
338-3040. Our office is in-service and closed to
the public on Wednesdays.

SF State is an Equal Opportunity/Americans with
Disabilities Act employer and has a strong
commitment to the principles of diversity.



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From: nb-at-dada.no
Date: Tue, 19 Oct 2010 01:47:31 -0500
Subject: [Microscopy] How to transport an SEM ?

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

Hi
Not so wise to sniff glutaraldehyde but I would lke to ask you if
de-activation of glutaraldehyde after long storage is accompanied by
reduction of its characteristic smell.
Best
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
eikonika-at-otenet.gr
www.aim.cat
*********************************

l
----- Original Message -----
X-from: {FMonson-at-wcupa.edu}
To: {eikonika-at-otenet.gr}
Sent: Monday, October 18, 2010 8:59 PM

Regarding " reduction of its characteristic smell ".

The simple answer is yes.

'Smell' is not always odor, however.

"In a more recent study by Cain, et al., it was shown that there was some
odor detection even below 0.0001 ppm, though the point of 50% detection,
commonly used as the criterion for threshold, occurred at 0.0003 ppm for
glutaraldehyde. The results implied some chemesthetic activity in the range
0.035 to 0.100 ppm, but failed to follow a concentration-response
relationship characteristic of chemesthesis. The term chemesthesis refers
to perception of feel from chemicals regardless of whether subjects would be
able to notice the sensory quality - pleasant or unpleasant, irritating or
nonirritating (Caine, 2007), Gaffney 2007}. They concluded that the
chemesthetic detection at hundreds of parts per billion (ppb) found in brief
exposures does not occur at concentrations as low as 0.1 ppm for longer
exposures. The functions for ocular detection and nasal localization imply
some perception of feel a little below 0.2 ppm, although the points of 50%
detection occurred at 0.39 and 0.47 ppm for the eye and the nose,
respectively."

Hence glutaraldehyde vapors can be perceived as present at very low
concentrations, 50% of those exposed at 0.0003 ppm, and could cause
irritation around 0.035-0.100 ppm. Some chemicals, such as the aldehydes
formaldehyde and acetaldehyde, have shown irritation thresholds at 3-10
times odor thresholds(Schiffman, 2000). Other chemicals, such as the sulfur
compounds, have shown irritation thresholds at 10^3to 10^4 times odor
thresholds (Schiffman, 2000). In general, the majority of volatile organic
compounds elicit chemesthesis at an order of magnitude above odor thresholds
(Cain, 2007; Cometto, 2004)"


Cain, William S., Roland Schmidt, and Alfredo Jalowayski: Odor and
Chemesthesis from Exposures to Glutaraldehyde Vapor. International Archives
of Occupational and Environmental Health Int Arch Occup Environ Health 80:
721-731, 2007.

Cometto-Muniz, et al.: Detection of single and mixed VOCs by smell and by
sensory irritation, Indoor Air, 14(S8):108-117, 2004.

Gaffney, 2007 Gaffney, Shannon H., and Dennis Paustenbach: A Proposed
Approach for Setting Occupational Exposure Limits for Sensory Irritants
Based on Chemosensory Models. Ann Occ Hyg, 51, 4, 345-356, 2007.

Schiffman, Susan S., et al.: Potential Health Effects of Odor from Animal
Operations, Wastewater Treatment, and Recycling of Byproducts. Journal of
Agromedicine 7(1):7-82. 2000.



Tony

Ps Note: This is from an article I'm about to submit for publication.

......................................................................
Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC 5250 E US 36, Suite 830
Avon IN 46123 www.ph2llc.com
(317) 718-7020 off
(317) 718-7038 fax
(317) 409-3238 cell

90% of Risk Management is knowing where to place the decimal point...any
consultant can give you the other 10%(SM)


-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: Monday, October 18, 2010 8:13 PM
To: ph2-at-sprynet.com

Hi
Not so wise to sniff glutaraldehyde but I would lke to ask you if
de-activation of glutaraldehyde after long storage is accompanied by
reduction of its characteristic smell.
Best
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
eikonika-at-otenet.gr
www.aim.cat
*********************************

l
----- Original Message -----
X-from: {FMonson-at-wcupa.edu}
To: {eikonika-at-otenet.gr}
Sent: Monday, October 18, 2010 8:59 PM

Hi all,

I'm rescuing an old JEOL 6400 with EDS, it is working fine where it is
now at a institute, but I have to take it home this week and has made
a room for it in my cellar. But how do I disconnect and transport it
in a proper manner?
If anyone has any advice on this I'm very pleased to know. I didn’t
find any advice in the manuals.

Thanks in advance
Nils Berner
nb-at-dada.no

Åsveien 26
1369 Stabekk
NORWAY

Tlf +47 67123030
Mob +47 92866366


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From: cpawlowicz-at-ubmtechinsights.com
Date: Tue, 19 Oct 2010 14:50:05 -0500
Subject: [Microscopy] Oxford EDS help

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

Thank you Tony for this acurate information, the numbers are impressive. So,
we cannot judge by the smell if glutar is still ok
Good luck with your submission
yorgos


----- Original Message -----
X-from: {ph2-at-sprynet.com}
To: {eikonika-at-otenet.gr}
Sent: Tuesday, October 19, 2010 4:53 AM

We've successfully rescued a Zeiss Leo 1455VP SEM (bought cheap at gov't auction, DOA) with some excellent help from people on this list (thanks!).

We're now turning our attention to the attached EDS system which came with it (Oxford EDS, Inca Energy software).

It came missing most of the cables, a smashed window, missing collimator/electron trap, but did include the main assembly (LN2 dewar, detector), two blue boxes (Inca Xstream and Inca MICS), Inca software and a bunch of license key disks.

After gathering various cables and bits we've built up a PC with winXP and installed Inca. We bought the DB37 cable from Oxford but sourced the other ones ourselves (firewire, coax etc). Before getting a new window installed we wanted to see if the detector was alive - pumped it down overnight, filled with LN2, and tried to fire it up.

Inca is able to talk to the SEM PC via rs-232, that seems ok (it can see and change mag/kv/beam conditions) but fails when trying to capture an image through INCA (comes up with an error 'This system has hardware which is not INCA compatible').

It can also see things like the LN2 level/ temperature. If we try to run a scan we get a hardware error.

Failed to start xray acquisition - failed to access device - the requested device was not found - error 3012 (4)

Any helpful hints / ideas / things to try?

thanks,

Chris Pawlowicz, B.Eng
Manager, Lab Development
UBM TechInsights
3000 Solandt Rd Ottawa ON Canada K2K 2X2
T: +1 613 576 0150
F: +1 613 599 6501
E: cpawlowicz-at-ubmtechinsights.com
W: www.ubmtechinsights.com


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From: raristau-at-ims.uconn.edu
Date: Tue, 19 Oct 2010 16:02:52 -0500
Subject: [Microscopy] Re: How to transport an SEM ?

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

Nils,

Your situation reminded me of a great story I found on-line. A gentleman
from Sweden bought a used TEM and moved it to his home. He kept a running
journal of the experience, which I think anyone in a similar situation would
find interesting, if not helpful. Follow to this site:

http://www.home.neab.net/gandalf/EM-lab/TEM100CX/

Good luck!

Roger

Roger A. Ristau, PhD
Electron Microscopy Specialist
Institute of Materials Science
97 North Eagleville Road
University of Connecticut
Storrs, CT 06269
vox: 860-486-5453
fax: 860-486-4745


} From: {nb-at-dada.no}
} Reply-To: {nb-at-dada.no}
} Date: Tue, 19 Oct 2010 01:51:50 -0500
} To: {raristau-at-ims.uconn.edu}
} Subject: [Microscopy] How to transport an SEM ?
}
}
}
}
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} Hi all,
}
} I'm rescuing an old JEOL 6400 with EDS, it is working fine where it is
} now at a institute, but I have to take it home this week and has made
} a room for it in my cellar. But how do I disconnect and transport it
} in a proper manner?
} If anyone has any advice on this I'm very pleased to know. I didn‚t
} find any advice in the manuals.
}
} Thanks in advance
} Nils Berner
} nb-at-dada.no
}
} Ã…sveien 26
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} NORWAY
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From: kraftpiano-at-gmail.com
Date: Tue, 19 Oct 2010 16:31:28 -0500
Subject: [Microscopy] SEM value.

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

Dear listers,

I am in the unfortunate position of having to provide a value estimate for a used SEM for a shipping damage claim. It's an older model, so I was hoping some of you wouldn't mind sharing your purchase prices or sales prices for a JEOL JSM-35C, turbo-pumped with basic configuration (Only SE and BE detectors). The microscope arrived damaged beyond repair, and I need to provide an approximate value for the instrument.

Please respond off-line.

Thank you,

Justin A. Kraft

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From: werner1-at-slb.com
Date: Wed, 20 Oct 2010 09:59:47 -0500
Subject: [Microscopy] Any experiences with "Dino-Lite" digital microscope?

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

Listers,

I know that "you get what you pay for - if you're lucky" so am quite ambivalent. Saw this demoed at a trade show, can't believe the low price (~$500), seeking feedback.

Their web site is http://bigc.com/ and the product is a small "handheld" digital microscope camera - 10x-50x basically - comes with software. It looks 'way too good to be true, but if it is even half as capable as it seemed, it could replace my Polaroid MP-4 (no more Type 52 film) for macros.

If anyone has any experience with this instrument, or comments, I'd truly appreciate your input. Thanks in advance.

Regards,
Andrew Werner
Chief Metallurgist, Perforating
Schlumberger Reservoir Completions
14910 Airline Road
Rosharon, TX 77583




==============================Original Headers==============================
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8, 39 -- Subject: Any experiences with "Dino-Lite" digital microscope?
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From: vray-at-partbeamsystech.com
Date: Wed, 20 Oct 2010 10:41:53 -0500
Subject: [Microscopy] Re: Any experiences with "Dino-Lite" digital microscope?

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

I am using a 1.3MP variant of this microscope, purchased off E-bay about
a year ago without any brand names on it. Using it to inspect Si surface
of polished backside samples and occasional soldering of SMT components
- it is adequate for these purposes. Image is surprisingly clear and
colors are reasonably close to reality. Some kind of stand is a "must"
at high magnification.

The only complain I have is long-term stability of the focusing
mechanism - once picture is in focus I have to snap it within 5 to 10
seconds or it may drift slightly. But then again, my device is a generic
clone and most likely was not made to Dino-Lite standards.

If in doubt - go to E-Bay, search for "USB microscope", get similar
device for $30 and play with it. If it seems close to what you need in
terms of magnification and clarity then invest in Dino-Lite with more
confidence, if not then you left with a nice toy for your kids :)

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com

On 10/20/2010 11:01 AM, werner1-at-slb.com wrote:
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
}
} Listers,
}
} I know that "you get what you pay for - if you're lucky" so am quite ambivalent. Saw this demoed at a trade show, can't believe the low price (~$500), seeking feedback.
}
} Their web site is http://bigc.com/ and the product is a small "handheld" digital microscope camera - 10x-50x basically - comes with software. It looks 'way too good to be true, but if it is even half as capable as it seemed, it could replace my Polaroid MP-4 (no more Type 52 film) for macros.
}
} If anyone has any experience with this instrument, or comments, I'd truly appreciate your input. Thanks in advance.
}
} Regards,
} Andrew Werner
} Chief Metallurgist, Perforating
} Schlumberger Reservoir Completions
} 14910 Airline Road
} Rosharon, TX 77583
}
}
}
}
} ==============================Original Headers==============================
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From: ph2-at-sprynet.com
Date: Wed, 20 Oct 2010 12:14:39 -0500
Subject: [Microscopy] Re: Any experiences with "Dino-Lite" digital

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

Valery,

Thank you!

I received similar feedback from two other listmembers, but failed to cc the list when replying to them.

This information is quite valuable - not good to spend more than I need to, but neither do I want to buy something "cheap" that will not do the job. The demo was convincing but it really seemed too-good-to-be-true. Experience of others so far substantiates that it is useful.

I'll look on e-bay as you suggest - many uses at home, from pulling splinters from fingers to cleaning hearing-aid earpieces - to "check it out".

Thanks again.

Regards,
Andrew

-----Original Message-----
X-from: vray-at-partbeamsystech.com [mailto:vray-at-partbeamsystech.com]
Sent: Wednesday, October 20, 2010 10:42 AM
To: Andrew Werner
Cc: microscopy-at-microscopy.com

I purchased one in 2008 from Think Geek at $455.99 including shipping (I
have no financial interest in them).

For basic images it's fair.

Mine came with a stand that held fine for focusing.

The biggest problem is adjusting the lighting for even feedback - it is very
much similar to vignetting. A quick BW threshold on an image reveals the
poor response.

It is often quite noticeable even without that.

There is also some spherical aberration and depth of focus is poor at the
outer annular space.

I don't use it for anything where these are issues, e.g, good reports.

But it was only $460 and I can move it around very easily.


Tony

-----Original Message-----
X-from: vray-at-partbeamsystech.com [mailto:vray-at-partbeamsystech.com]
Sent: Wednesday, October 20, 2010 11:45 AM
To: ph2-at-sprynet.com

I am using a 1.3MP variant of this microscope, purchased off E-bay about
a year ago without any brand names on it. Using it to inspect Si surface
of polished backside samples and occasional soldering of SMT components
- it is adequate for these purposes. Image is surprisingly clear and
colors are reasonably close to reality. Some kind of stand is a "must"
at high magnification.

The only complain I have is long-term stability of the focusing
mechanism - once picture is in focus I have to snap it within 5 to 10
seconds or it may drift slightly. But then again, my device is a generic
clone and most likely was not made to Dino-Lite standards.

If in doubt - go to E-Bay, search for "USB microscope", get similar
device for $30 and play with it. If it seems close to what you need in
terms of magnification and clarity then invest in Dino-Lite with more
confidence, if not then you left with a nice toy for your kids :)

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com

On 10/20/2010 11:01 AM, werner1-at-slb.com wrote:
}
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}
} Listers,
}
} I know that "you get what you pay for - if you're lucky" so am quite
ambivalent. Saw this demoed at a trade show, can't believe the low price
(~$500), seeking feedback.
}
} Their web site is http://bigc.com/ and the product is a small "handheld"
digital microscope camera - 10x-50x basically - comes with software. It
looks 'way too good to be true, but if it is even half as capable as it
seemed, it could replace my Polaroid MP-4 (no more Type 52 film) for macros.
}
} If anyone has any experience with this instrument, or comments, I'd truly
appreciate your input. Thanks in advance.
}
} Regards,
} Andrew Werner
} Chief Metallurgist, Perforating
} Schlumberger Reservoir Completions
} 14910 Airline Road
} Rosharon, TX 77583
}
}
}
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From: bozhilov-at-ucr.edu
Date: Wed, 20 Oct 2010 13:46:27 -0500
Subject: [Microscopy] SCSMM meeting Nov.9th

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

The Southern California Society for Microscopy and Microanalysis (SCSMM) invites you to
attend the forthcoming Fall Meeting at the UCLA campus.

For directions and further details visit SCSMM web site: www.scsmm.org
2010 Fall Meeting
Tuesday, November 9th, 2010
University of California at Los Angeles

Agenda:
---------------------------------------------------------------------
4:00 p.m. Demo sessions on Thermo Scientific fully integrated EDS,
WDS, EBSD analytical system ,
Room 1230 Engineering V building, UCLA

6:00 p.m. Hot Buffet Dinner served in the CNSI building, UCLA
(sponsored by Thermo Sci, Physical Electronics USA, Zeiss Inc. and FEI Co.).

Cost $10 (regular), Students $4 - which cover the SCSMM membership dues for 2010/11 as well.

7:00 p.m. CNSI Auditorium
William Schopf, Director of UCLA's Center for the Study of Evolution and the Origin of Life,
Department of Earth and Space Sciences, UCLA
“The Earliest History of Life: Solution to Darwin's Dilemma."

7:45 p.m. John Callaghan, Physical Electronics USA
“Gas Cluster Ion Beam for Chemical Depth Profiling of Organic Materials”

7:55 p.m. John Konopka, Thermo Scientific, USA
“Fully Integrated EDS, WDS, EBSD Analysis”

8:05 p.m. Dan Jacobson, Carl Zeiss Inc.
"Carl Zeiss Analytical Power for the Sub-Nanometer World"

8:15 p.m. Amanda Englund, FEI company, TBA

8:25 p.m. Best SEM Image MNA and FEI Student Award Presentations
_________________________________________________________
ADVANCED RESERVATION IS REQUIRED!

Respond no later than 5 p.m. Tuesday, November 2nd, 2010
Contact: Jim Kuleck at (818) 354 5666, email: james.kulleck-at-jpl.nasa.gov.

If interested to schedule individual demos on the Thermo Sci. EDS, EBSD, WDS system during
the week of Nov. 8th-12th at the UCLA campus please contact:
Wayne Watson, 650 969 2273, wayne.watson-at-thermofisher.com.




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From: rcommon-at-msu.edu
Date: Wed, 20 Oct 2010 14:36:56 -0500
Subject: [Microscopy] glutaraldehyde dilution

Contents Retrieved from Microscopy Listserver Archives
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Amanda,

Well, I have now.

And now I'm confused... but that's good!

One or the other seems like just what I need to replace the MP-4 for macros.

Time for further research.

Thanks!

Andrew

-----Original Message-----
X-from: Amanda Lawrence [mailto:ALawrence-at-entomology.msstate.edu]
Sent: Wednesday, October 20, 2010 11:47 AM
To: Andrew Werner

Valery,

Thank you!

I received similar feedback from two other listmembers, but failed to cc the list when replying to them.

This information is quite valuable - not good to spend more than I need to, but neither do I want to buy something "cheap" that will not do the job. The demo was convincing but it really seemed too-good-to-be-true. Experience of others so far substantiates that it is useful.

I'll look on e-bay as you suggest - many uses at home, from pulling splinters from fingers to cleaning hearing-aid earpieces - to "check it out".

Thanks again.

Regards,
Andrew

-----Original Message-----
X-from: vray-at-partbeamsystech.com [mailto:vray-at-partbeamsystech.com]
Sent: Wednesday, October 20, 2010 10:42 AM
To: Andrew Werner
Cc: microscopy-at-microscopy.com

As someone who worked in an EM service lab for many years, I am well aware of the problem of having people "dilute glutaraldehyde with buffer". If you dilute 8% glutaraldehyde with an equal amount of 0.2M buffer you get 4% glutaraldehyde in 0.1M buffer, which is fine. But many times clients diluted 25% or 50% glutaraldehyde with 0.2M buffer only, resulting in a strongly hypertonic solution which causes the tissue to appear very dense, obscuring ultrastructural detail and causing poor sectioning. Glutaraldehyde should be diluted to twice the desired end concentration with water, then mixed with an equal volume of 0.2M buffer. I also remember reading somewhere that concentrated glutaraldehyde is more stable than dilute.

Ralph Common



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I am not sure what you mean by "Fact is, it must be diluted in water and not in buffer..." - Clearly I dilute my glutaraldehyde into the buffer I use.

Unlike you, I have a strong recollection of reading that 8% glutaraldehyde was more stable than more concentrated versions but can't find any reference for this statement.



Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


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From: leunissen-at-aurion.nl
Date: Wed, 20 Oct 2010 15:00:04 -0500
Subject: [Microscopy] aldehydes and buffer concentration

Contents Retrieved from Microscopy Listserver Archives
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Dear list

I have never quite understood the relevance of the concentration of the buffer components in glutaraldehyde or formaldehyde fixatives.
No doubt there is an effect of the buffer, this has been amply illustrated. But if one considers the contribution of the aldehydes to the molarity of the fixative solution, it seems something else than tonicity is at play.

Monomeric glutaraldehyde has a Mwt of 100. A 4% solution corresponds to 0.4M.

The situation is even more strange for formaldehyde with a Mwt of 30. A 4% solution being 1.33M.

Clearly at the very onset of fixation the situation is that the fixative is highly hypertonic.
Perhaps it is rather the buffer capacity than the osmolarity that is important?

Always keen to learn..

Jan Leunissen

Aurion




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From: PhillipsT-at-missouri.edu
Date: Wed, 20 Oct 2010 15:07:02 -0500
Subject: [Microscopy] aldehydes and buffer concentration

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My understanding is that they can cross the membrane and therefore don't count in the osmolarity. Tom


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: leunissen-at-aurion.nl [mailto:leunissen-at-aurion.nl]
Sent: Wednesday, October 20, 2010 3:01 PM
To: Phillips, Thomas E.

Dear list

I have never quite understood the relevance of the concentration of the buffer components in glutaraldehyde or formaldehyde fixatives.
No doubt there is an effect of the buffer, this has been amply illustrated. But if one considers the contribution of the aldehydes to the molarity of the fixative solution, it seems something else than tonicity is at play.

Monomeric glutaraldehyde has a Mwt of 100. A 4% solution corresponds to 0.4M.

The situation is even more strange for formaldehyde with a Mwt of 30. A 4% solution being 1.33M.

Clearly at the very onset of fixation the situation is that the fixative is highly hypertonic.
Perhaps it is rather the buffer capacity than the osmolarity that is important?

Always keen to learn..

Jan Leunissen

Aurion




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From: NEERAJG-at-clemson.edu
Date: Wed, 20 Oct 2010 15:57:56 -0500
Subject: [Microscopy] Re: 103rd Annual Meeting of the National Shellfisheries

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Dear Colleagues,

We are pleased to announce  two special sessions; 1) Biofouling  and 2) Shell Formation (Biomineralization) at the 103rd  Annual Meeting of the National Shellfisheries Association (NSA) in March 27-31 2011 in Baltimore, Maryland. (http://shellfish.org/ )


1) We anticipate the Biofouling session to be broad and multidisciplinary focusing on various aspects of marine and fresh water biofouling including but not limited to imaging. Since imaging is relevant to this community, we welcome abstracts on,

A) Imaging studies of fouling organism highlighting cellular and developmental processes
B) Tracking fouling organisms, development of lab and field assays for evaluating new coatings
B) Being multidisciplinary, we also welcome abstracts from folks who are working on the materials aspect of biofouling by engineering new and improved antifouling coatings and polymers, specially characterization of such materials using various microscopy techniques and or biological assays. 

We also welcome abstracts on other aspects of biofouling such as impact of invasive fouling species etc. I will be chairing the Biofouling session so please feel free to contact me if you have any questions, my contact details are in my signature.
 

2) Similarly we anticipate the Shell Formation (Biomineralization) session to be broad and multidisciplinary as well. We welcome abstracts on various aspects of Biomineralization including but not limited to,

A) Cellular and developmental processes involved in Biomineralization
B) Characterization of shells and or other biomineralized structures using various microscopy techniques
C) Physical, biological and  or chemical aspects of the Biomineralization process
D) Effects of ocean acidification on shell formation and biomineralization

We would also like to invite abstracts from the Bone formation community, folks who are working on the comparative cellular and developmental aspects of bone formation and biomineralization.

If you have any questions about the Shell Formation (Biomineralization) session please contact my colleague 

Dr. Andrew S. Mount
Associate Professor
Okeanos Research Group
Department of Biological Sciences
132 Long Hall
Clemson University
Clemson, SC-29631
USA
Email: mount-at-clemson.edu
Phone: 864-656-3597


Please forward this announcement to your friends and collaborators who are working in these areas. More information on the 103rd Annual Meeting of NSA can be found at http://shellfish.org/  


Best Regards,

Neeraj V. Gohad, Ph.D.
Postdoctoral Fellow
Okeanos Research Group
Department of Biological Sciences
132 Long Hall
Clemson University
Clemson,SC-29634
USA
Email: neerajg-at-clemson.edu  
Phone: 864-656-3597

Website: http://www.clemson.edu/okeanos    












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From: leunissen-at-aurion.nl
Date: Wed, 20 Oct 2010 17:23:50 -0500
Subject: [Microscopy] Re: aldehydes and buffer concentration

Contents Retrieved from Microscopy Listserver Archives
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Thanks, Tom.

Small and chemically relatively inert molecules might be able to cross the membrane, but wouldn't that only be possible if the molecules were not to interact with the membrane? Or where interaction is only weak and reversible?
The aldehydes are extremely reactive and covalently fix membrane components, both proteins and some lipids and some carbohydrates. As soon as a membrane is exposed to fixative the membrane will lose its semipermeability, facilitating penetration of fixative and buffer. Even larger molecules such as antibodies and cell components can under circumstances pass fixed membranes. Your comment triggered the rusty brain and made me realise that the hypertonicity will perhaps not be experienced and have a serious effect since exposure to the hypertonic medium also implies fixation has kicked in at the very same moment. However, once semipermeability has been lost one wouldn't think the osmolarity of the buffer is still relevant anymore.

Back to square one and still wondering..


Jan

On 21/10/2010, at 9:06 AM, Phillips, Thomas E. wrote:

} My understanding is that they can cross the membrane and therefore don't count in the osmolarity. Tom
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://www.biology.missouri.edu/faculty/phillips.html
} http://www.biotech.missouri.edu/mcc/
}
} -----Original Message-----
} From: leunissen-at-aurion.nl [mailto:leunissen-at-aurion.nl]
} Sent: Wednesday, October 20, 2010 3:01 PM
} To: Phillips, Thomas E.
} Subject: [Microscopy] aldehydes and buffer concentration
}
}
}
}
} ----------------------------------------------------------------------------
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} Dear list
}
} I have never quite understood the relevance of the concentration of the buffer components in glutaraldehyde or formaldehyde fixatives.
} No doubt there is an effect of the buffer, this has been amply illustrated. But if one considers the contribution of the aldehydes to the molarity of the fixative solution, it seems something else than tonicity is at play.
}
} Monomeric glutaraldehyde has a Mwt of 100. A 4% solution corresponds to 0.4M.
}
} The situation is even more strange for formaldehyde with a Mwt of 30. A 4% solution being 1.33M.
}
} Clearly at the very onset of fixation the situation is that the fixative is highly hypertonic.
} Perhaps it is rather the buffer capacity than the osmolarity that is important?
}
} Always keen to learn..
}
} Jan Leunissen
}
} Aurion
}
}
}
}
} ==============================Original Headers==============================
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From: clei-at-illinois.edu
Date: Wed, 20 Oct 2010 19:19:44 -0500
Subject: [Microscopy] viaWWW: Gatan SmartSet Model 901

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Email: clei-at-illinois.edu
Name: changhui Lei

Organization: University of Illinois

Title-Subject: [Filtered] Gatan SmartSet Model 901

Message: Dear Colleagues,

We're using the Gatan SmartSet Model 901 Hot Stage Controller. This
particular one has a 12-volt DC water pump. When the temperature of
the stage goes above 500C, water cooling is activated.

Our unit is intermittent in operation of the water pump. We've
noticed that when the pump runs, output voltage applied to the pump
is about 2.4 to 2.5 volts DC. When the controller won't run the
pump, the output is only about 1.2 volts DC - not enough to drive the
motor. We have also measured the open circuit pump output. Not
knowing the circuit however, this isn't meaningful yet.

Note that it's quite simple to test this without expending the time
and effort of heating up the holder. When the "specimen holder"
cable is disconnected from the controller, the controller thinks that
the measured temperature is very high, so it activates the pump
output. When you turn on the controller, you'll see the measured
temperature ramp up; as it passes 500C the pump should run.

When the pump runs, it's clear that it works (pumps water).

Has anyone seen this problem and come up with a solution? Can you
verify the drive voltage to the pump for us (with pump on and open)?


Any feedback is welcome.

Changhui


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From: vakimler-at-med.wayne.edu
Date: Wed, 20 Oct 2010 19:21:06 -0500
Subject: [Microscopy] viaWWW: Steric Hindrance with Copper Reagents, Grid and

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State Univ. School of Medicine

Title-Subject: [Filtered] Steric Hindrance with Copper Reagents, Grid
and Immunogold Labeling

Message: We have tried some labeling of nuclei for thin-section TEM
with our primary antibody, the Click-It EdU reagents (containing
copper) and fluoronanogold, which we silver-enhanced to ~20 nm.

It appears that we did not get specific labeling and so-called
"dirty" sections. Some gold was within the nucleus and in small
clusters or individuals, but also throughout the cell.

We have heard that copper grids cannot be used for immunogold
labeling. It is possible that the copper sulfate from the EdU reagent
kit may have created some steric hindrance with the gold or some
other feature of these two metals render them ineffective if used
together?

I wanted to add that we used nickel-asbestos finder grids, not copper
ones. The copper was in the EdU reagent.

Thanks,
Vickie

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From: gary-at-gaugler.com
Date: Wed, 20 Oct 2010 19:27:48 -0500
Subject: [Microscopy] Re: viaWWW: Gatan SmartSet Model 901

Contents Retrieved from Microscopy Listserver Archives
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Not knowing the circuit, it sounds like a power supply
problem. Check the circuit board for a three terminal
regulator like a 7812. If it is there, check side pins.
One is input and the other is output. If input is greater
than 12V and output is not 12V then this device has failed.
Replace it. If input is not greater than 12V, then the
input bridge rectifier or capacitor are suspect. Depending
on how old the unit is, the input or output filter electrolytic
capacitors will dry out and either be open or in this case,
shorted. That would engage the regulator short circuit
protection mode.

gary g.


At 05:21 PM 10/20/2010, you wrote:



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From: leunissen-at-aurion.nl
Date: Wed, 20 Oct 2010 20:13:14 -0500
Subject: [Microscopy] Re: viaWWW: Steric Hindrance with Copper Reagents, Grid and

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Hello again, Vickie


Copper grids (which you are not using) are usually avoided in immunolabeling as Cu-ions sometimes react with buffers during prolonged incubations and cause precipitates over the grid surface. This has been topic of discussion on this list, if I am not mistaken. You may want to check the archives.
Many amino acids have chelating properties, i.e. they can bind Cu-ions. In theory this could modify the epitope surface and thereby poison antigen-antibody interactions, but as far as I know this has never been documented. It would at least be prudent to wash well after using reagents that contain heavy metal ions before applying any type of immuno reagents.
Negatively charged gold may result in background in the nucleus as a result of charge based interactions, but your reaction may be positive. What do your negative controls look like? Signal/Noise ratios can usually be controlled with a proper protein block and incubation buffer composition. It is hard to give proper advice without details of what your incubations were like. Feel free to contact me off list to discuss details.

Good luck,


Jan Leunissen

AURION - ImmunoGold Reagents
http://www.aurion.nl

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From: allan.mitchell-at-stonebow.otago.ac.nz
Date: Wed, 20 Oct 2010 20:29:29 -0500
Subject: [Microscopy] Electron microscopy technical position available

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Hi all

An opportunity exists for an experienced electron microscopy
technician to work in the Otago Centre for Electron Microscopy (OCEM)
at the University of Otago, Dunedin, New Zealand. This position has
two principle roles. The appointee will be required to support our
expanding SEM Section. The other principal role is to develop,
implement and promote correlative light and transmission electron
microscopy techniques. On occasions, and as time permits, the
appointee will also be required to provide back-up technical support
in conventional transmission electron microscopy.

The successful applicant will have an enthusiasm for electron
microscopy with experience in either scanning electron microscopy and/
or transmission electron microscopy. Previous experience in
immunofluorescence and/or immunocytochemistry is highly desirable.
The appointee will be expected to successfully train users of the
Centre in these techniques and keep up to date with developments in
these areas. The ability to work as part of a team, cope with a wide
range of demands, work without direct supervision, set priorities, and
bring enthusiasm to the job, is essential.

If you know of someone who may be suitable please bring this to their
attention.

A copy of the Job description, Application Form and Equal Employment
Opportunity (EEO) Form can be found at the web address; http://www.otago.ac.nz/vacancies/general/otago014452.html
, or I can send you copies if this suits better (my email is below).

Specific enquiries may be directed to Mr Allan Mitchell, Manager of
the Otago Centre for Electron Microscopy, email; allan.mitchell-at-stonebow.otago.ac.nz

Applications quoting reference number G10/593 close on Wednesday 17
November 2010.

For information about OCEM visit our website - http://ocem.otago.ac.nz/

Regards

Allan
Allan Mitchell
Microscopy Otago - Electron Microscopy
c/- Department of Anatomy and Structural Biology
Otago School of Medical Sciences
P.O. Box 913
Dunedin
New Zealand

Phone (03) 479 5642 or 479 7301
Fax (03) 479 5086 or 479 7254

EM Centre: http://ocem.otago.ac.nz/
Confocal Centre: http://occm.otago.ac.nz/
NZ Microscopy Society: http://microscopynz.co.nz/




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From: allan.mitchell-at-stonebow.otago.ac.nz
Date: Wed, 20 Oct 2010 23:05:47 -0500
Subject: [Microscopy] Cryostages on SEM

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

Hi all

We are throwing around some ideas for a cryo-experiment and we want
to try a couple of experiments. We have stored away some old SEM cryo-
stages, one of which we are going to use. However before commit
ourselves I need some basic data. Does anybody know the operational
temperature range of the following:

EmiTech K1250 SEM cold stage (1990's vintage)

Hexland SEM cold stage (1980's vintage)

EMscope SP2000 SEM cold stage (1980's vintage)

Thanks

Allan


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From: protrain-at-emcourses.com
Date: Thu, 21 Oct 2010 07:19:39 -0500
Subject: [Microscopy] Cryostages on SEM

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

Hi Allan

In my day of designing cryo systems we worked on the principle, probably
from freeze fracture experts, that each interface between the liquid
nitrogen and the specimen was a loss of 10 degrees.

I was deeply involved with the SP2000 and the K1250 both of which
substantiated the 10 degree figure.

Good luck with the systems.

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com


-----Original Message-----
X-from: allan.mitchell-at-stonebow.otago.ac.nz
[mailto:allan.mitchell-at-stonebow.otago.ac.nz]
Sent: 21 October 2010 05:07
To: protrain-at-emcourses.com

Hi all

We are throwing around some ideas for a cryo-experiment and we want
to try a couple of experiments. We have stored away some old SEM cryo-
stages, one of which we are going to use. However before commit
ourselves I need some basic data. Does anybody know the operational
temperature range of the following:

EmiTech K1250 SEM cold stage (1990's vintage)

Hexland SEM cold stage (1980's vintage)

EMscope SP2000 SEM cold stage (1980's vintage)

Thanks

Allan


==============================Original Headers==============================
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8, 19 -- Subject: Cryostages on SEM
8, 19 -- Date: Thu, 21 Oct 2010 17:05:43 +1300
8, 19 -- X-Mailer: Apple Mail (2.936)
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From: FMonson-at-wcupa.edu
Date: Thu, 21 Oct 2010 08:41:43 -0500
Subject: [Microscopy] glutaraldehyde dilution

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

I remember an abstract from an early 20th century meeting in which the author announced that "Formalin" - out of the bottle - was the best fixative for something or other or everything - I read it 'before Xerox" while 'browsing the stacks'.

While working in urology research during the 1990's - age-related research for me - I read an article concerning the use of DMSO to 'treat' irritable bladder syndrome in which the author quipped that 50% DMSO was an excellent histologic fixative for the urothelium (transitional epithelium of the excretory system).

Proof that if one reads enough, one will have the opportunity to become discouraged.

Cheers,

Fred

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
Schmucker Science Center South
West Chester University, West Chester, PA 19383
610-738-0437

Home Page: http://cmirt.wcupa.edu
Scheduler: http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl


-----Original Message-----
X-from: rcommon-at-msu.edu [mailto:rcommon-at-msu.edu]
Sent: Wednesday, October 20, 2010 3:44 PM
To: Monson, Frederick

As someone who worked in an EM service lab for many years, I am well aware of the problem of having people "dilute glutaraldehyde with buffer". If you dilute 8% glutaraldehyde with an equal amount of 0.2M buffer you get 4% glutaraldehyde in 0.1M buffer, which is fine. But many times clients diluted 25% or 50% glutaraldehyde with 0.2M buffer only, resulting in a strongly hypertonic solution which causes the tissue to appear very dense, obscuring ultrastructural detail and causing poor sectioning. Glutaraldehyde should be diluted to twice the desired end concentration with water, then mixed with an equal volume of 0.2M buffer. I also remember reading somewhere that concentrated glutaraldehyde is more stable than dilute.

Ralph Common



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

I am not sure what you mean by "Fact is, it must be diluted in water and not in buffer..." - Clearly I dilute my glutaraldehyde into the buffer I use.

Unlike you, I have a strong recollection of reading that 8% glutaraldehyde was more stable than more concentrated versions but can't find any reference for this statement.



Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


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26, 27 -- From FMonson-at-wcupa.edu Thu Oct 21 08:41:43 2010
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From: kenconverse-at-qualityimages.biz
Date: Thu, 21 Oct 2010 10:04:26 -0500
Subject: [Microscopy] How to transport an SEM ?

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

Hi Nils,
The 6400 is a nice instrument.

Of course, make sure all the connections are properly labeled (confirmed
when disconnecting). Notes can be helpful.

1)Remove the EDS detector before moving the optics console.
2)Remove the HT tank before moving the electronics console.
3)Bolt down the floating table on the optics console. Hopefully you've got
the lifting bolts from its original delivery. If not, at least 4 M16-2.0 x
40 bolts will do the trick.
4)Keep as much of the system under vacuum as possible. Height constraints
may require the removal of the gun, condenser lens and vacuum standpipe in
the rear, but if you can leave V7 closed (and in place) you can keep the
chamber and stage under vacuum. If it has a turbo pump, remove that before
moving the optics console.
5)The electronics and power consoles are on wheels. No problem to move, but
the optics console is going to need to be lifted and provided with wheels.
Safe jacks (Raise- n-Roll or other) are great, but can be hard to find. A
pallet truck or even a strong dolly will also work. A pry bar (Johnson Bar)
and some wood will get things off the ground.

Best wishes for its happy new home.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: nb-at-dada.no [mailto:nb-at-dada.no]
Sent: Tuesday, October 19, 2010 2:50 AM
To: kenconverse-at-qualityimages.biz

Hi all,

I'm rescuing an old JEOL 6400 with EDS, it is working fine where it is
now at a institute, but I have to take it home this week and has made
a room for it in my cellar. But how do I disconnect and transport it
in a proper manner?
If anyone has any advice on this I'm very pleased to know. I didn’t
find any advice in the manuals.

Thanks in advance
Nils Berner
nb-at-dada.no

Åsveien 26
1369 Stabekk
NORWAY

Tlf +47 67123030
Mob +47 92866366


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From: jacques.faerber-at-ipcms.u-strasbg.fr
Date: Thu, 21 Oct 2010 10:42:38 -0500
Subject: [Microscopy] Noran Vantage Xserver error

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

Hi all

We have with a probleme on our old but good working EDS analyser, a
Noran Vantage. Good working until yesterday, when the hard drive failed
after 10 years good services.
We put a new drive, and install a ghost image of the system we had on
CDs, and much is working again but not all, not the usefullest. The
spectrum and imaging application Vista (not MS Vista ! Noran Vista !)
works ok. But as I try to start Spectral Display, or Image Display, or
Easy Micro, I get an error message :
"X Toolkit Erro: can't open display: :0.0".
There is something wrong with XViev or the X server, but I have no idea
how it works, and what is to be done. I remember we soon had this
message once, but the probleme resolved from itself.

Does someone have a Vantage running, and some idea what to do ?

Thanks in advance

Jacques

--
J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mailJacques.Faerber-at-ipcms.u-strasbg.fr


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From: hkonishi-at-wisc.edu
Date: Thu, 21 Oct 2010 13:14:57 -0500
Subject: [Microscopy] Diffraction Rings Distortion-Analysis

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

Hello,

I downloaded the script and I am trying to use it with my Digital Micrograph. Nothing appeared on my Digital Micrograph. My DM version is 1.71.38 . If anyone use the script, please advise me if I can use the script with my DM. I tried to reach the author, but his mail I know is not working.

Thank you,

Hiromi
UW-Maqdison

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From: leunissen-at-aurion.nl
Date: Thu, 21 Oct 2010 13:39:23 -0500
Subject: [Microscopy] Re: aldehydes and buffer concentration

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Thank you all for your contributions. I learned a lot again.
The paper by Arborgh et al answered many questions!


Jan Leunissen
Aurion

On 22/10/2010, at 4:44 AM, Geoff McAuliffe wrote:

} See Arbrogh, et al. J. Ultrastruc. Res. 56:339, 1976.
}
} Geoff
}
}
} leunissen-at-aurion.nl wrote:
} } ----------------------------------------------------------------------------
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} } Dear list
} }
} } I have never quite understood the relevance of the concentration of the buffer components in glutaraldehyde or formaldehyde fixatives. No doubt there is an effect of the buffer, this has been amply illustrated. But if one considers the contribution of the aldehydes to the molarity of the fixative solution, it seems something else than tonicity is at play.
} } Monomeric glutaraldehyde has a Mwt of 100. A 4% solution corresponds to 0.4M.
} } The situation is even more strange for formaldehyde with a Mwt of 30. A 4% solution being 1.33M.
} }
} } Clearly at the very onset of fixation the situation is that the fixative is highly hypertonic.
} } Perhaps it is rather the buffer capacity than the osmolarity that is important?
} }
} } Always keen to learn..
} }
} } Jan Leunissen
} }
} } Aurion
} }
} }
} }
} } ==============================Original Headers==============================
} } 10, 21 -- From leunissen-at-aurion.nl Wed Oct 20 15:00:03 2010
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} } 10, 21 -- Subject: [Microscopy] aldehydes and buffer concentration
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} } 10, 21 -- Date: Thu, 21 Oct 2010 08:59:51 +1300
} } 10, 21 -- Cc: Ralph Common {rcommon-at-msu.edu}
} } 10, 21 -- Message-Id: {11BA2A49-80DB-4B44-BD61-337F56D4A591-at-aurion.nl}
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} }
} }
} }
}
}
} --
} --
} **********************************************
} Geoff McAuliffe, Ph.D.
} Neuroscience and Cell Biology
} Robert Wood Johnson Medical School
} 675 Hoes Lane, Piscataway, NJ 08854
} voice: (732)-235-4583 mcauliff-at-umdnj.edu
} **********************************************
}
}



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From: david.knecht-at-uconn.edu
Date: Thu, 21 Oct 2010 15:04:26 -0500
Subject: [Microscopy] Hemocytometers

Contents Retrieved from Microscopy Listserver Archives
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We need a new hemocytometer and in shopping around, have two questions:
1. Fisher sells two different ones: "Reichert* Bright-Line Counting Chambers are molded from quartz Pyrex* glass; Hausser Bright-Line Counting Chambers are milled from soda lime glass". Does anyone know why you would buy Pyrex vs. soda lime glass for nearly the same price?
2. For about 1/5 the price of either, you can now buy a Chinese knockoff. Has anyone tried these?
Thanks- Dave

Dr. David Knecht
Department of Molecular and Cell Biology
Co-head Flow Cytometry and Confocal Microscopy Facility
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)




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From: martimor-at-nmsu.edu
Date: Thu, 21 Oct 2010 15:40:32 -0500
Subject: [Microscopy] RE: Leica / Reichert-Jung 2030 Biocut Microtome

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Does anyone use a Leica / Reichert-Jung 2030 Biocut Microtome? I am
looking for some information on this piece of equipment. If anyone can
help, I would greatly appreciate it!

Kind regards,
Marti Morales-Ensign



==============================Original Headers==============================
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From: axelsson-at-acc.umu.se
Date: Fri, 22 Oct 2010 02:49:25 -0500
Subject: [Microscopy] How to transport an SEM ?

Contents Retrieved from Microscopy Listserver Archives
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Not only do I have the website, I'm also on this mailing list.... :-)

Three general tips for moving microscopes.

1. Use a digital camera and document every stage of dismantling,
especially if you never have moved a microscope before. I took close to
700 pictures when moving the TEM and I had to use one to get it hooked
up right.

2. Any vacuum connections is wrapped in Aluminium foil. Al doesn't emit
fibres and is easily formed and stays on.

3. Don't use tape, use plastic foil to bind together cables and other
stuff, it doesn't leave any glue. Tape could leave glue... don't ask! In
the end wrap the whole microscope, it keeps cables from getting tangled
in door handles and prevents dust during the move.

Nils, I realise that it's a bit late now, but if you want to call and
ask me some questions, my number is +46 73 98 67 881.

PS. Anyone got an EDS for a JEOL JSM-25? I got one a year ago but
haven't taken time to add its story to my site yet.

Good luck!

/Göran

raristau-at-ims.uconn.edu wrote:
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
}
} Nils,
}
} Your situation reminded me of a great story I found on-line. A gentleman
} from Sweden bought a used TEM and moved it to his home. He kept a running
} journal of the experience, which I think anyone in a similar situation would
} find interesting, if not helpful. Follow to this site:
}
} http://www.home.neab.net/gandalf/EM-lab/TEM100CX/
}
} Good luck!
}
} Roger
}
} Roger A. Ristau, PhD
} Electron Microscopy Specialist
} Institute of Materials Science
} 97 North Eagleville Road
} University of Connecticut
} Storrs, CT 06269
} vox: 860-486-5453
} fax: 860-486-4745
}
}
}
} } From: {nb-at-dada.no}
} } Reply-To: {nb-at-dada.no}
} } Date: Tue, 19 Oct 2010 01:51:50 -0500
} } To: {raristau-at-ims.uconn.edu}
} } Subject: [Microscopy] How to transport an SEM ?
} }
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} } ----------------------------------------------------------------------------
} }
} } Hi all,
} }
} } I'm rescuing an old JEOL 6400 with EDS, it is working fine where it is
} } now at a institute, but I have to take it home this week and has made
} } a room for it in my cellar. But how do I disconnect and transport it
} } in a proper manner?
} } If anyone has any advice on this I'm very pleased to know. I didn‚t
} } find any advice in the manuals.
} }
} } Thanks in advance
} } Nils Berner
} } nb-at-dada.no
} }
} } Ã…sveien 26
} } 1369 Stabekk
} } NORWAY
} }
} } Tlf +47 67123030
} } Mob +47 92866366
} }
} }


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From: johnf-at-geology.wisc.edu
Date: Fri, 22 Oct 2010 08:31:22 -0500
Subject: [Microscopy] problem with Hitachi S3400 W filaments

Contents Retrieved from Microscopy Listserver Archives
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We have been struggling for many months trying to understand why we
are having premature failure of Hitachi S3400 W filaments. We have
plotted lifetimes of filaments vs batches and have seen a marked
decline in life of filaments. Have any other Hitachi SEM users seen
this behavior. Not only that, we see two very strange modes of
failures of the filaments: either fracture (not thinning or melting)
very close to the tip, with _rough_ breakage surfaces (again, not
melting or thinning), and/or _thinning 2/3 of the way down the legs
to the posts.

John Fournelle
--
========================================================
John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480
Cameron Electron Microprobe Lab lab: (608) 265-4798
Department of Geoscience fax: (608) 262-0693
University of Wisconsin home: (608) 274-2245
1215 West Dayton St. email: johnf-at-geology.wisc.edu
Madison, WI 53706 amateur radio: WA3BTA
Personal http://www.geology.wisc.edu/~johnf/
Probe lab http://www.geology.wisc.edu/~johnf/sx51.html
Probe Sign Up Calender:
http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi

"The first rule of all intelligent tinkering is to save every cog and
wheel." -- Aldo Leopold

"For a successful technology, reality must take precedence over
public relations, for Nature cannot be fooled." -- Richard P.
Feynman

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From: rcsencsits-at-lbl.gov
Date: Fri, 22 Oct 2010 11:51:33 -0500
Subject: [Microscopy] Decommissioning Zeiss EM10

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In a couple of weeks we are decommissioning our vintage orange Zeiss EM10 circa 1976. I am not sure of the paperwork but it will be available as a unit or in pieces. Moving expenses are on you. For the last 5 years we have run at 80kV; it is used daily.

Any interested takers please email me.
Roseann

Roseann Csencsits, PhD
Manager Donner TEM Facility
Lawrence Berkeley Lab 01-365
1 Cyclotron Road
Berkeley, CA 94720
510-486-4548











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From: Georgianne.Ciraolo-at-cchmc.org
Date: Mon, 25 Oct 2010 08:12:58 -0500
Subject: [Microscopy] viaWWW: soft blocks

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Title-Subject: [Filtered] soft blocks

Message: Does anyone have a suggestion on what to do with blocks that
are to soft to cut? These were embed in LX-112 and for some reason
they did not harden. Is there anyway that this experiment can be
salvaged? Thanks in advance

Georgianne Ciraolo
Dept of Pathology
Cincinnati Children's Hospital

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From: TindallR-at-missouri.edu
Date: Mon, 25 Oct 2010 08:57:46 -0500
Subject: [Microscopy] viaWWW: soft blocks

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Gergianne,

Ah, the dreaded "gummy bears". I am not familiar with this particular resin, but I don't know of any way to save samples that have incompletely polymerized and refuse to polymerize further with standard EM resins. For us, it's pretty much a write-off.

If there actually is a method of saving blocks like this, I would love to know it...

Good luck,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




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Title-Subject: [Filtered] soft blocks

Message: Does anyone have a suggestion on what to do with blocks that
are to soft to cut? These were embed in LX-112 and for some reason
they did not harden. Is there anyway that this experiment can be
salvaged? Thanks in advance

Georgianne Ciraolo
Dept of Pathology
Cincinnati Children's Hospital

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From: dcromey-at-email.arizona.edu
Date: Mon, 25 Oct 2010 09:36:12 -0500
Subject: [Microscopy] viaWWW: soft blocks

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Georgianne,

I worked with LX-112 a long time ago on peripheral nerve tissue. Sometimes
there was enough attached fat on the sample that it impeded good
polymerization. Depending on how soft your blocks are, I have sometimes had
good luck putting some of the catalyst (I no longer remember the name for
this component, but it's usually the smallest bottle) on a swab, smearing it
on the surface (already trimmed and faced) and putting the block back in the
oven overnight. It tends to firm up that surface enough for decent
sections, as long as you don't go in too far from the surface.

Doug

^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Douglas W. Cromey, M.S. - Assistant Scientific Investigator
Dept. of Cell Biology & Anatomy, University of Arizona
1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA

office: AHSC 4212 email: Cromey-at-Arizona.edu
voice: 520-626-2824 fax: 520-626-2097

http://swehsc.pharmacy.arizona.edu/exppath/
Home of: "Microscopy and Imaging Resources on the WWW"

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Title-Subject: [Filtered] soft blocks

Message: Does anyone have a suggestion on what to do with blocks that
are to soft to cut? These were embed in LX-112 and for some reason
they did not harden. Is there anyway that this experiment can be
salvaged? Thanks in advance

Georgianne Ciraolo
Dept of Pathology
Cincinnati Children's Hospital

Login Host: 205.142.197.75
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From: W.Muss-at-salk.at
Date: Mon, 25 Oct 2010 09:41:13 -0500
Subject: [Microscopy] Re: Soft blocks

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Dear Georgianne,
(dear all),

assisting Randy Tindall's opinion/posting: no way / no method to save the non-polymerized blocks as to my knowledge too.

I have embedded diagnostic tissue(s) in LX-112 (via PropOxide as well as Acetonitrile as an intermedium) without any problems so far for the last 20 years, using the same weight formula(s) as for (old) Epon 812 and some substitute expox-resin (e.g. glycidether 100 from Serva). LX-112 being a good stuff (disclaimer: only satisfied consumer client, no financial interest in Ladd Res.Int,USA).

Concerning {reasons} , why "gummi(y)-bears": manifold.... starting at least in the dehydration sequence...e. g.
also using old, perhaps deteriorated PO, or deteriorated resin component, like old catalyst (using DMP-30? or another one); insufficient mixing of complete resin or "forgotten" catalyst which can happen....

Sorry about the meager/slim future aspect of / fat chance for your specimens.....

And last but not least: If anybody knows of a method to rescue such unpolymerized or not fully polymerized blocks I would love to know about it, too. Thank you!

Best wishes and regards

Wolfgang MUSS
EM-Lab, Pathology
SALK-LKH (Gen.Hosp.)
Salzburg, Austria



} -----Ursprüngliche Nachricht-----
} Von: Georgianne.Ciraolo-at-cchmc.org [mailto:Georgianne.Ciraolo-at-cchmc.org]
} Gesendet: Montag, 25. Oktober 2010 15:18
} An: Muß Wolfgang
} Betreff: [Microscopy] Soft blocks [embedding with LX-112, do not/did
} not harden...]
}
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} Email: Georgianne.Ciraolo-at-cchmc.org
} Name: Georgianne Ciraolo
} Organization: Cincinnati Children's Hospital
} Title-Subject: soft blocks
} Message: Does anyone have a suggestion on what to do with blocks that
} are to soft to cut?
}
} These were embed in LX-112 and for some reason they did not harden. Is
} there anyway that this experiment can be
} salvaged?
}
} Thanks in advance
}
} Georgianne Ciraolo
} Dept of Pathology
} Cincinnati Children's Hospital
}
}
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From: thoward-at-unm.edu
Date: Mon, 25 Oct 2010 10:12:14 -0500
Subject: [Microscopy] Re: viaWWW: soft blocks

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

Georgianne -

I've had this happen once or twice, & have been desperate
(& curious) enough to take blocks back down to propylene
oxide or acetone to try to recover samples that ended up
in gummi bear-type resin. Trim the excess resin from
around your tissue pieces and then put the tissue back
into your final solvent - use a fresh bottle, since I'm
assuming incomplete dehydration was the problem. If you
went straight from ethanol to resin, you'll probably need
to go to one of the traditional "transitional" solvents
(PO, acetone, whatever). Ethanol-to-resin isn't a problem
if you are careful, but I don't think you can remove the
semi-polymerized resin very well with just EtOH. I've
never tried, though.... Go through a couple of changes of
the solvent (time depends on how many samples you
have/vial, tissue volume, etc.), then re-infiltrate with
fresh resin (several changes) & hope for the best! Doesn't
always give great results, but if they are one-of-a-kind
samples it is worth a try.

For the record, I usually don't keep resin components more
than a year once the bottle is opened & I try to be really
careful about keeping the final solvent water-free.

Good luck!

Tamara

On Mon, 25 Oct 2010 08:14:52 -0500
Georgianne.Ciraolo-at-cchmc.org wrote:
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} Name: Georgianne Ciraolo
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} Organization: Cincinnati Children's Hospital
}
} Title-Subject: [Filtered] soft blocks
}
} Message: Does anyone have a suggestion on what to do
} with blocks that
} are to soft to cut? These were embed in LX-112 and for
} some reason
} they did not harden. Is there anyway that this
} experiment can be
} salvaged? Thanks in advance
}
} Georgianne Ciraolo
} Dept of Pathology
} Cincinnati Children's Hospital
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***************************
Tamara Howard
Cell Biology & Physiology
UNM-HSC
Albuquerque, NM
***************************


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From: kenconverse-at-qualityimages.biz
Date: Mon, 25 Oct 2010 10:13:57 -0500
Subject: [Microscopy] problem with Hitachi S3400 W filaments

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

Hi John,
I think there is a possibility that there are 2 separate problems.

1) The fractures may be caused by a bad batch of W wire. This happens from
time to time, I am told.

2) The thinning way down the legs may be a vacuum problem. I've found that
having a very small leak in the gun of most instruments shortens filament
life more than a large leak in the chamber. I think this is due to the
creation of localized high pressure areas. You may have a small leak in the
insulator that the wehnelt is mounted on, producing very high pressures
inside the wehnelt.

Perhaps others with direct knowledge of the S3400 will chime in.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: johnf-at-geology.wisc.edu [mailto:johnf-at-geology.wisc.edu]
Sent: Friday, October 22, 2010 9:34 AM
To: kenconverse-at-qualityimages.biz

We have been struggling for many months trying to understand why we
are having premature failure of Hitachi S3400 W filaments. We have
plotted lifetimes of filaments vs batches and have seen a marked
decline in life of filaments. Have any other Hitachi SEM users seen
this behavior. Not only that, we see two very strange modes of
failures of the filaments: either fracture (not thinning or melting)
very close to the tip, with _rough_ breakage surfaces (again, not
melting or thinning), and/or _thinning 2/3 of the way down the legs
to the posts.

John Fournelle
--
========================================================
John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480
Cameron Electron Microprobe Lab lab: (608) 265-4798
Department of Geoscience fax: (608) 262-0693
University of Wisconsin home: (608) 274-2245
1215 West Dayton St. email: johnf-at-geology.wisc.edu
Madison, WI 53706 amateur radio: WA3BTA
Personal http://www.geology.wisc.edu/~johnf/
Probe lab http://www.geology.wisc.edu/~johnf/sx51.html
Probe Sign Up Calender:
http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi

"The first rule of all intelligent tinkering is to save every cog and
wheel." -- Aldo Leopold

"For a successful technology, reality must take precedence over
public relations, for Nature cannot be fooled." -- Richard P.
Feynman

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4, 24 -- Subject: problem with Hitachi S3400 W filaments
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==============================Original Headers==============================
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From: protrain-at-emcourses.com
Date: Mon, 25 Oct 2010 11:18:30 -0500
Subject: [Microscopy] problem with Hitachi S3400 W filaments

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

Hi All

I think Ken is absolutely correct. The wire is just simply tungsten wire it
is not produced specially for electron microscope filament production.

This wire sounds over brittle to me according to the type of fracture,
perhaps a material scientist could tell us what embrittles tungsten; even a
EDX analysis may answer that?

The other possible solution mentioned, a poor vacuum, would produce the
following clues

1. Thinning over the complete filament not just near the tip
2. Short filament life
3. An oily ozone smell in the gun chamber
4. Yellow/orange filament ceramic contamination

Most of all do not be fooled by and vacuum indicator as the gauge is likely
to be well away from the gun area. The gun is a fast pumping area and the
best way to recognise this would be to compare penning and pirani values.
Fast pumping areas have reasonable penning values monitored on the column
but poor pirani values at the backing pump. The easy flow from the high
pumping area records as reasonable but the excess gas gives the backing pump
a hard time.

Good luck

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz]
Sent: 25 October 2010 16:15
To: protrain-at-emcourses.com

Hi John,
I think there is a possibility that there are 2 separate problems.

1) The fractures may be caused by a bad batch of W wire. This happens from
time to time, I am told.

2) The thinning way down the legs may be a vacuum problem. I've found that
having a very small leak in the gun of most instruments shortens filament
life more than a large leak in the chamber. I think this is due to the
creation of localized high pressure areas. You may have a small leak in the
insulator that the wehnelt is mounted on, producing very high pressures
inside the wehnelt.

Perhaps others with direct knowledge of the S3400 will chime in.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: johnf-at-geology.wisc.edu [mailto:johnf-at-geology.wisc.edu]
Sent: Friday, October 22, 2010 9:34 AM
To: kenconverse-at-qualityimages.biz

We have been struggling for many months trying to understand why we
are having premature failure of Hitachi S3400 W filaments. We have
plotted lifetimes of filaments vs batches and have seen a marked
decline in life of filaments. Have any other Hitachi SEM users seen
this behavior. Not only that, we see two very strange modes of
failures of the filaments: either fracture (not thinning or melting)
very close to the tip, with _rough_ breakage surfaces (again, not
melting or thinning), and/or _thinning 2/3 of the way down the legs
to the posts.

John Fournelle
--
========================================================
John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480
Cameron Electron Microprobe Lab lab: (608) 265-4798
Department of Geoscience fax: (608) 262-0693
University of Wisconsin home: (608) 274-2245
1215 West Dayton St. email: johnf-at-geology.wisc.edu
Madison, WI 53706 amateur radio: WA3BTA
Personal http://www.geology.wisc.edu/~johnf/
Probe lab http://www.geology.wisc.edu/~johnf/sx51.html
Probe Sign Up Calender:
http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi

"The first rule of all intelligent tinkering is to save every cog and
wheel." -- Aldo Leopold

"For a successful technology, reality must take precedence over
public relations, for Nature cannot be fooled." -- Richard P.
Feynman

==============================Original Headers==============================
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==============================Original Headers==============================
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From: ehaller-at-health.usf.edu
Date: Mon, 25 Oct 2010 12:32:53 -0500
Subject: [Microscopy] viaWWW: soft blocks

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

Hi, Georgianne,

As a last resort, you can try to polymerize the blocks at 90 degrees C for a few hours, or even overnight, and try cutting them after they cool. Sometimes this will harden the surface of the blocks enough to allow a few sections. Cut a few sections thicker than usual, pick them up on coated grids if you need to, and see if you can use them. Cut smaller block faces than usual also. You will also need to cut faster than usual to get sections. I've cut blocks with the consistency of bubblegum before, or pencil erasers (a researcher gave me 30 year old blocks that had been in someone's attic) by popping them in a hot oven for a few hours, then cutting just the first few microns at a speed faster than I've cut any thins before. It worked, he got his photos, and was a happy camper.

Ed Haller

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-cas.usf.edu
________________________________________
X-from: Georgianne.Ciraolo-at-cchmc.org [Georgianne.Ciraolo-at-cchmc.org]
Sent: Monday, October 25, 2010 9:24 AM
To: Haller, Edward

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: Georgianne.Ciraolo-at-cchmc.org
Name: Georgianne Ciraolo

Organization: Cincinnati Children's Hospital

Title-Subject: [Filtered] soft blocks

Message: Does anyone have a suggestion on what to do with blocks that
are to soft to cut? These were embed in LX-112 and for some reason
they did not harden. Is there anyway that this experiment can be
salvaged? Thanks in advance

Georgianne Ciraolo
Dept of Pathology
Cincinnati Children's Hospital

Login Host: 205.142.197.75
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From: swalck-at-southbaytech.com
Date: Mon, 25 Oct 2010 13:38:00 -0500
Subject: [Microscopy] problem with Hitachi S3400 W filaments

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

Just a little tutorial about Tungsten. Tungsten will embrittle when it is
heated over the last processing temperature. When it is being drawn, the
manufacturers are careful not to heat it over the previous temperature at
which it was drawn. As soon as you turn on a tungsten filament, you will
heat it above its last processing temperature, so it will be brittle.

When tungsten is used as a filament, a small amount of thoria is added to
the mix. What this does it pin the grain boundaries from excessive grain
growth while it is hot. Without the thoria, the grains will grow and you
can actually see them offset along the wire. The grains will appear to have
sheared along the grain boundaries of these large grains and is usually the
failure mechanism when the thoria is not present. With the thoria present,
the grains stay small and the wire gets progressively smaller as it
evaporates at a slow rate within the gun. You will see the wire smaller at
the failure point.

A poor vacuum will also cause tungsten filaments to fail. Look for a yellow
oxide telltale. An overheated filament that evaporates will have a more
bluish color of evaporated material on the surrounding supports.

-Scott

Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA 92673

eMail: SWalck-at-SouthBayTech.com
Phone: 949-492-2600
Toll Free: 1-800-728-2233 (USA)
Fax: 949-492-1499

www.SouthBayTech.com



-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: Monday, October 25, 2010 9:29 AM
To: swalck-at-southbaytech.com

Hi All

I think Ken is absolutely correct. The wire is just simply tungsten wire it
is not produced specially for electron microscope filament production.

This wire sounds over brittle to me according to the type of fracture,
perhaps a material scientist could tell us what embrittles tungsten; even a
EDX analysis may answer that?

The other possible solution mentioned, a poor vacuum, would produce the
following clues

1. Thinning over the complete filament not just near the tip
2. Short filament life
3. An oily ozone smell in the gun chamber
4. Yellow/orange filament ceramic contamination

Most of all do not be fooled by and vacuum indicator as the gauge is likely
to be well away from the gun area. The gun is a fast pumping area and the
best way to recognise this would be to compare penning and pirani values.
Fast pumping areas have reasonable penning values monitored on the column
but poor pirani values at the backing pump. The easy flow from the high
pumping area records as reasonable but the excess gas gives the backing pump
a hard time.

Good luck

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz]

Sent: 25 October 2010 16:15
To: protrain-at-emcourses.com

Hi John,
I think there is a possibility that there are 2 separate problems.

1) The fractures may be caused by a bad batch of W wire. This happens from
time to time, I am told.

2) The thinning way down the legs may be a vacuum problem. I've found that
having a very small leak in the gun of most instruments shortens filament
life more than a large leak in the chamber. I think this is due to the
creation of localized high pressure areas. You may have a small leak in the
insulator that the wehnelt is mounted on, producing very high pressures
inside the wehnelt.

Perhaps others with direct knowledge of the S3400 will chime in.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: johnf-at-geology.wisc.edu [mailto:johnf-at-geology.wisc.edu]
Sent: Friday, October 22, 2010 9:34 AM
To: kenconverse-at-qualityimages.biz

We have been struggling for many months trying to understand why we
are having premature failure of Hitachi S3400 W filaments. We have
plotted lifetimes of filaments vs batches and have seen a marked
decline in life of filaments. Have any other Hitachi SEM users seen
this behavior. Not only that, we see two very strange modes of
failures of the filaments: either fracture (not thinning or melting)
very close to the tip, with _rough_ breakage surfaces (again, not
melting or thinning), and/or _thinning 2/3 of the way down the legs
to the posts.

John Fournelle
--
========================================================
John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480
Cameron Electron Microprobe Lab lab: (608) 265-4798
Department of Geoscience fax: (608) 262-0693
University of Wisconsin home: (608) 274-2245
1215 West Dayton St. email: johnf-at-geology.wisc.edu
Madison, WI 53706 amateur radio: WA3BTA
Personal http://www.geology.wisc.edu/~johnf/
Probe lab http://www.geology.wisc.edu/~johnf/sx51.html
Probe Sign Up Calender:
http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi

"The first rule of all intelligent tinkering is to save every cog and
wheel." -- Aldo Leopold

"For a successful technology, reality must take precedence over
public relations, for Nature cannot be fooled." -- Richard P.
Feynman

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From: tina-at-pbrc.hawaii.edu
Date: Mon, 25 Oct 2010 13:38:33 -0500
Subject: [Microscopy] Re: viaWWW: soft blocks

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

I've used LX-112 for years, love it, and have rarely had problems with it.
So saying, I now have gummy bears in the oven! I suspect the catalyst is
the culprit in this case. Often when it gets old, addition of it to the
resin makes the resin darker orange. In this case the resin didn't darken
at all, which made me suspicious. After two days in the oven at 60C I was
still getting fingerprints int he surface. I turned the oven up to about
75C, which helped make them harder to the fingernail, so we'll see.

In addition to upping the temperture, I have also known people to try UV
and, in one desperate case, one friend froze the blocks and quickly
sectioned them before they thawed. I can't remember if it was -20C, -40C,
or colder, but too cold and they would be brittle.

Wish us both luck!
Aloha,
Tina


} Message: Does anyone have a suggestion on what to do with blocks that
} are to soft to cut? These were embed in LX-112 and for some reason
} they did not harden. Is there anyway that this experiment can be
} salvaged? Thanks in advance
}
} Georgianne Ciraolo
} Dept of Pathology
} Cincinnati Children's Hospital
}
} Login Host: 205.142.197.75
} ---------------------------------------------------------------------------
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} 7, 22 -- Subject: viaWWW: soft blocks
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}

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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From: allan.mitchell-at-stonebow.otago.ac.nz
Date: Mon, 25 Oct 2010 14:14:19 -0500
Subject: [Microscopy] soft blocks

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

Hi all

On occasions we have had soft blocks we have been able to rescue them
by trimming away most the gooey resin, soaking in propylene oxide for
an hour or 2 (if the tissue is robust enough you can apparently
sonicate for a short period in propylene oxide (Bauman and Mendell
1974) but I have never done this. After soaking in propylene oxide
re-infiltrate with increasing resin mixture in propylene oxide again
followed by polymerisation. This has worked for us in most cases.

This method came from Principles and Techniques of Electron
Microscopy, Biological Applications, by M.A Hayat (3rd Edition) page
133. Bauman and Mendells paper is from Stain Technology Vol 49, pp
119 (1974). They were using Spurrs, we have used this method with
some success with Agar 100 epoxy resin.

Good luck

Allan





Allan Mitchell
Microscopy Otago - Electron Microscopy
c/- Department of Anatomy and Structural Biology
Otago School of Medical Sciences
P.O. Box 913
Dunedin
New Zealand

Phone (03) 479 5642 or 479 7301
Fax (03) 479 5086 or 479 7254

EM Centre: http://ocem.otago.ac.nz/
Confocal Centre: http://occm.otago.ac.nz/
NZ Microscopy Society: http://microscopynz.co.nz/




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From: mls1-at-ebsciences.com
Date: Mon, 25 Oct 2010 16:42:04 -0500
Subject: [Microscopy] problem with Hitachi S3400 W filaments

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

John,

Ken and Steve are correct. The W wire we use is not specific to EM Filaments, however, the quality/purity is critical. And despite the material certifications that we filament manufacturers require from our suppliers, we occasionally do get a questionable batch. This makes Customer feedback a critical component of our quality control efforts. If we hear from a Customer who is having trouble with filament life and there is even a remote chance that the it is a quality issue, we follow-up with other Customers who have used tips from the same lot. So if you go back to your supplier, be it us (EBS) or otherwise, they could have information that can help you. We have not heard of any problems with our Hitachi filaments in recent memory.

Generally speaking, if the problem is with the W material, your filament life would have diminished with the first bad filament you installed and subsequent tips from the same lot would have yielded results very similar to the first one. However, if the degradation in life has been progressive over time, that would be more indicative of a vacuum issue.

I hope this helps.

Best regards,

Michael R. Nesta
President
Energy Beam Sciences, Inc.
Tel: 860 653-0411
Fax: 860 653-0422
www.ebsciences.com
"Adding Brilliance to Your Vision"


-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: Monday, October 25, 2010 12:27 PM
To: mls1

Hi All

I think Ken is absolutely correct. The wire is just simply tungsten wire it
is not produced specially for electron microscope filament production.

This wire sounds over brittle to me according to the type of fracture,
perhaps a material scientist could tell us what embrittles tungsten; even a
EDX analysis may answer that?

The other possible solution mentioned, a poor vacuum, would produce the
following clues

1. Thinning over the complete filament not just near the tip
2. Short filament life
3. An oily ozone smell in the gun chamber
4. Yellow/orange filament ceramic contamination

Most of all do not be fooled by and vacuum indicator as the gauge is likely
to be well away from the gun area. The gun is a fast pumping area and the
best way to recognise this would be to compare penning and pirani values.
Fast pumping areas have reasonable penning values monitored on the column
but poor pirani values at the backing pump. The easy flow from the high
pumping area records as reasonable but the excess gas gives the backing pump
a hard time.

Good luck

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz]
Sent: 25 October 2010 16:15
To: protrain-at-emcourses.com

Hi John,
I think there is a possibility that there are 2 separate problems.

1) The fractures may be caused by a bad batch of W wire. This happens from
time to time, I am told.

2) The thinning way down the legs may be a vacuum problem. I've found that
having a very small leak in the gun of most instruments shortens filament
life more than a large leak in the chamber. I think this is due to the
creation of localized high pressure areas. You may have a small leak in the
insulator that the wehnelt is mounted on, producing very high pressures
inside the wehnelt.

Perhaps others with direct knowledge of the S3400 will chime in.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: johnf-at-geology.wisc.edu [mailto:johnf-at-geology.wisc.edu]
Sent: Friday, October 22, 2010 9:34 AM
To: kenconverse-at-qualityimages.biz

We have been struggling for many months trying to understand why we
are having premature failure of Hitachi S3400 W filaments. We have
plotted lifetimes of filaments vs batches and have seen a marked
decline in life of filaments. Have any other Hitachi SEM users seen
this behavior. Not only that, we see two very strange modes of
failures of the filaments: either fracture (not thinning or melting)
very close to the tip, with _rough_ breakage surfaces (again, not
melting or thinning), and/or _thinning 2/3 of the way down the legs
to the posts.

John Fournelle
--
========================================================
John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480
Cameron Electron Microprobe Lab lab: (608) 265-4798
Department of Geoscience fax: (608) 262-0693
University of Wisconsin home: (608) 274-2245
1215 West Dayton St. email: johnf-at-geology.wisc.edu
Madison, WI 53706 amateur radio: WA3BTA
Personal http://www.geology.wisc.edu/~johnf/
Probe lab http://www.geology.wisc.edu/~johnf/sx51.html
Probe Sign Up Calender:
http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi

"The first rule of all intelligent tinkering is to save every cog and
wheel." -- Aldo Leopold

"For a successful technology, reality must take precedence over
public relations, for Nature cannot be fooled." -- Richard P.
Feynman

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From: speransv-at-mail.nih.gov
Date: Mon, 25 Oct 2010 16:59:57 -0500
Subject: [Microscopy] Re: viaWWW: soft blocks

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

One more vote for putting blocks in a freezer.
Long ago and far away, we used to do that. And it did help.

Now I know that those blocks were softish because they were not polymerized right: there was a mysterious tradition teaching to put the resin first into 37C for 1 day, then into 60C. While maybe not quite as gummy as the candy store gummy bears, those blocks just kept yielding to the screw when you tried to secure them in the holder!
Here, in the Land Of The Free, I learned to polymerize right, and never had this problem again.
(I do replace the accelerator often with a new bottle.)

Vlad
________________________________________________
Vlad Speransky, Staff Scientist
Biomedical Engineering and Physical Science Shared Resource
National Institute of Biomedical Imaging and Bioengineering, NIH
13 South Dr, Rm. 3N17 MSC 5766
Bethesda, MD 20892
301 496-3989
vladislav_speransky-at-nih.gov
http://www.nibib.nih.gov/Research/Intramural/SharedResource/Speransky

Opinions and experiences related are those of Vlad Speransky and do not represent the NIH. On the good side, this message is not confidential and can be freely shared and reproduced.

________________________________________
X-from: tina-at-pbrc.hawaii.edu [tina-at-pbrc.hawaii.edu]
Sent: Monday, October 25, 2010 2:51 PM
To: Speransky, Vlad (NIH/NIBIB) [E]

I've used LX-112 for years, love it, and have rarely had problems with it.
So saying, I now have gummy bears in the oven! I suspect the catalyst is
the culprit in this case. Often when it gets old, addition of it to the
resin makes the resin darker orange. In this case the resin didn't darken
at all, which made me suspicious. After two days in the oven at 60C I was
still getting fingerprints int he surface. I turned the oven up to about
75C, which helped make them harder to the fingernail, so we'll see.

In addition to upping the temperture, I have also known people to try UV
and, in one desperate case, one friend froze the blocks and quickly
sectioned them before they thawed. I can't remember if it was -20C, -40C,
or colder, but too cold and they would be brittle.

Wish us both luck!
Aloha,
Tina


} Message: Does anyone have a suggestion on what to do with blocks that
} are to soft to cut? These were embed in LX-112 and for some reason
} they did not harden. Is there anyway that this experiment can be
} salvaged? Thanks in advance
}
} Georgianne Ciraolo
} Dept of Pathology
} Cincinnati Children's Hospital
}
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}

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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14, 25 -- From speransv-at-mail.nih.gov Mon Oct 25 16:59:57 2010
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From: rpowell-at-nanoprobes.com
Date: Mon, 25 Oct 2010 19:07:00 -0500
Subject: [Microscopy] viaWWW: Steric Hindrance with Copper Reagents, Grid and

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Email: rpowell-at-nanoprobes.com
Name: Rick Powell

Organization: Nanoprobes, Incorporated

Title-Subject: [Filtered] Re: viaWWW: Steric Hindrance with Copper
Reagents, Grid and

Message: Hello Vickie:

Apologies for being late to the discussion...but there are a couple
more aspects of your system that it might be helpful to keep in mind:

(1) Rather than create steric hindrance, the copper may be acting as
a nucleating agent for silver deposition during the silver
enhancement step; thus you may be seeing a combination of specific
labeling with silver-enhanced gold, plus background silver deposition
triggered by copper ions. We have not studied copper, but other
redox-active metals can lead to silver deposition (see Gorm
Danscher's work for some examples and applications).

(2) You can remove transition metal ions using a chelating agent.
Disodium EDTA (ethylene diamine tetraacetic acid) is a good one as it
has a pH about 4.6 (we sometimes use 0.05 M on blots). We also found
that washing with sodium citrate buffer (0.02 M to 0.1 M) before the
silver enhancement gave us lower EM background than any other wash
buffer during early experiments with FluoroNanogold.

(3) Nonfat dried milk (1% up to 5%) in both the blocking and
incubation buffers is effective for controlling background,
especially with FluoroNanogold.

Hope this helps; feel free to contact me (off or on the list) if you
have more questions. More information on the incubations and washes
(as Jan points out) may let us provide more specific suggestions.

Regards,

Rick Powell

*****************************************************************************************
Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com

NANOPROBES, Incorporated
US Toll-free: (877) 447-6266 * Tel: (919) 845-6324 * Fax:
(631) 980-3608
*****************************************************************************************


Message: We have tried some labeling of nuclei for thin-section TEM
with our primary antibody, the Click-It EdU reagents (containing
copper) and fluoronanogold, which we silver-enhanced to ~20 nm.

It appears that we did not get specific labeling and so-called
"dirty" sections. Some gold was within the nucleus and in small
clusters or individuals, but also throughout the cell.

We have heard that copper grids cannot be used for immunogold
labeling. It is possible that the copper sulfate from the EdU reagent
kit may have created some steric hindrance with the gold or some
other feature of these two metals render them ineffective if used
together?

I wanted to add that we used nickel-asbestos finder grids, not copper
ones. The copper was in the EdU reagent.

Thanks,
Vickie



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From: Georgianne.Ciraolo-at-cchmc.org
Date: Mon, 25 Oct 2010 19:07:33 -0500
Subject: [Microscopy] [Filtered] MicroscopyListserverviaWWW: soft blocks- Thanks

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Email: Georgianne.Ciraolo-at-cchmc.org
Name: Georgianne Ciraolo

Organization: Cincinnati Children's Hospital

Title-Subject: [Filtered] soft blocks

Message:
Thanks to everyone for your good suggestions and encouragement for my
soft blocks question.

Have a great day

Regards

Georgianne

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From: lahernan-at-udec.cl
Date: Mon, 25 Oct 2010 21:01:23 -0500
Subject: [Microscopy] looking for WDS spectrometer for a JEOL 8600 probe

Contents Retrieved from Microscopy Listserver Archives
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Dear all,
I'm trying to add a new WDS spectrometer to our "earthquaked" JEOL JXA
8600 electron microprobe. Unfortunately these spectrometers are not longer
available at the factory so I wonder if there is somebody in the
Microscopy list who may have some offer.

If any, please contact me as soon as possible.

Best regards

Laura Hernandez



**********************************
MSc. Laura Hernandez
Laboratorio Microsonda Electronica
Instituto GEA
Universidad de Concepcion
Casilla 160c
Concepcion, 3
CHILE

TE +56-41-2204864 (oficina)
+56-41-2204861 (laboratorio)


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From: mark.grimson-at-ttu.edu
Date: Tue, 26 Oct 2010 19:41:07 -0500
Subject: [Microscopy] viaWWW: Service insurance v. OEM service contracts

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Email: mark.grimson-at-ttu.edu
Name: Mark Grimson

Organization: Texas Tech University

Title-Subject: [Filtered] Service insurance v. OEM service contracts

Message: Hello, an insurance equipment company will be visiting us in
the near future in an attempt to talk us into getting a policy
through them rather than the manufacturer's service contract, which
we have had for 40 years with great success and working relationship.
I am sure the insurance might be ok for some equipment, but not SEMs
and TEMs.

I am looking for any rationale I can use to help explain why the
Insurance route is not the way to go, and to stick with our
maintenance service contracts. Anecdotes or any experiences with
insurance on em's will be greatly appreciated. Thanks, Mark

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From: sgkcck-at-aol.com
Date: Tue, 26 Oct 2010 19:41:51 -0500
Subject: [Microscopy] viaWWW: Aurion Immunogold Workshop

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Email: sgkcck-at-aol.com
Name: Stacie Kirsch

Organization: Electron Microscopy Sciences

Title-Subject: [Filtered] Aurion Immunogold Workshop

Message: Electron Microscopy Sciences and Aurion are pleased to offer
a workshop on ImmunoGold Silver Staining at the House Ear Institute,
Los Angeles California from February 21-23, 2011. The course will
provide researchers with the opportunity to learn the theory and
practice of immunogold labeling while processing their own samples
under expert guidance.
The Immuno Gold Silver Staining Workshop is designed and taught by
Mr. Peter van de Plas who has been with Aurion since 1991. Mr. van de
Plas worked closely together with Dr. Leunissen in founding a firm
basis for Aurion including development of product applications. He
has been invited to many international microscopy conferences and
workshops and is especially experienced in providing hands-on
training. Dr. Paul Webster, head of Ahmanson Advanced EM & Imaging
Center at the at the House Ear Institute, will be providing expertise
in immunolabeling and applications of electron microscopy to
biomedical research.
The three day course will cover the properties of gold particles and
their protein conjugates, theories underlying immunogold labeling
protocols, and silver enhancement of gold particles. Also covered in
the course will be immunogold labeling on a variety of sample
preparations for LM, immunogold labeling for EM, pre/post-embedding
double immunogold labeling, and background minimization in immunogold
labeling.
Participants are encouraged to bring their own samples to maximize
learning. Please visit the course site for additional details:
http://www.emsdiasum.com/microscopy/products/immunogold/workshop.aspx
For more information contact
Stacie Kirsch
sgkcck-at-aol.com
215-412-8400



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From: rcsencsits-at-lbl.gov
Date: Tue, 26 Oct 2010 20:00:46 -0500
Subject: [Microscopy] Re: viaWWW: Service insurance v. OEM service contracts

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

Hi Mark,

With an insurance plan and I assume a pay-as-you-go service, you may find that the cost of replacements parts may be much higher than when under service contract. Typically when under contract the service engineer's time charges and the manufacturer's parts are at a discounted rate. Will insurance give you preventive visits as well as quick response for emergency visits?
If you machine is very old a third party service contract may work but I would not chance it on newer equipment.


Roseann Csencsits, PhD
Manager Donner TEM Facility
Lawrence Berkeley Lab 01-365
1 Cyclotron Road
Berkeley, CA 94720
510-486-4548









On Oct 26, 2010, at 5:53 PM, mark.grimson-at-ttu.edu wrote:

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} Title-Subject: [Filtered] Service insurance v. OEM service contracts
}
} Message: Hello, an insurance equipment company will be visiting us in
} the near future in an attempt to talk us into getting a policy
} through them rather than the manufacturer's service contract, which
} we have had for 40 years with great success and working relationship.
} I am sure the insurance might be ok for some equipment, but not SEMs
} and TEMs.
}
} I am looking for any rationale I can use to help explain why the
} Insurance route is not the way to go, and to stick with our
} maintenance service contracts. Anecdotes or any experiences with
} insurance on em's will be greatly appreciated. Thanks, Mark
}
}


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15, 32 -- Subject: Re: [Microscopy] viaWWW: Service insurance v. OEM service contracts
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From: vray-at-partbeamsystech.com
Date: Tue, 26 Oct 2010 21:02:25 -0500
Subject: [Microscopy] Re: viaWWW: Service insurance v. OEM service contracts

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

Hi Mark,

No insurance company have yet invented a magical way of up-keeping
complex particle beam instruments without the proper preventive
maintenance and qualified service. Neither have they a way of
outsourcing service to a vendor from central China or rural India
-at-US$2/Hr, to create money for covering there's own overhead expenses and
making profit without passing these extra costs to you, the end user.
One way or another, as direct expense and/or as cost of extended down
time when something serious happens to one of your instruments, service
insurance arrangement will ultimately get you into paying for overhead
expenses and profits of the insurance company - that is in addition to
the costs of parts, labor, travel, and overhead expenses of the service
organization (most likely the same OEM as you are using now).

I clearly understand why service insurance arrangement will good for the
equipment insurance company, but why and how could it possibly be good
for the end user is beyond my (limited) comprehension.

As institutional lab your safest bet is to stay with OEM service
contract, unless you and your boss are prepared to deal with political
implications of alternative approaches. If you are really desperate for
some cost reduction, then probably second safest bet is to get service
contract from one of reputable third-party SEM/TEM service providers,
there are few of them on this list.

Best of luck dealing with visit of Sirens :)

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com

On 10/26/2010 8:42 PM, mark.grimson-at-ttu.edu wrote:
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}
} Title-Subject: [Filtered] Service insurance v. OEM service contracts
}
} Message: Hello, an insurance equipment company will be visiting us in
} the near future in an attempt to talk us into getting a policy
} through them rather than the manufacturer's service contract, which
} we have had for 40 years with great success and working relationship.
} I am sure the insurance might be ok for some equipment, but not SEMs
} and TEMs.
}
} I am looking for any rationale I can use to help explain why the
} Insurance route is not the way to go, and to stick with our
} maintenance service contracts. Anecdotes or any experiences with
} insurance on em's will be greatly appreciated. Thanks, Mark
}
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From: gary-at-gaugler.com
Date: Wed, 27 Oct 2010 01:03:56 -0500
Subject: [Microscopy] Re: viaWWW: Service insurance v. OEM service

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

An insurance policy is nowhere near the same as a factory
maintenance contract, IMO. No way. I have never had an insurance
policy other than for natural hazards (but these can be covered)
and burglary, vandalism and business interruption and liability.

When talking about a SEM or TEM, it requires a very deep breath.
As Dirty Harry said, "Do you feel lucky?" OK, a HT tank fails
and needs replaced. A nominal $14K item but zero with a
maintenance contract. The odds of this happening? Greater
than zero. It has happened to me. Later model SEM HT tanks
do more than just HT...they do HT, filament current, extractor
current, SE HT and other high voltage sources. So there are
multiple points of failure here. Either one will take your tool
off-line. Without a contract, this will cost you time and
money. Oh, no contract? You are at the bottom of the
priority list. So good luck.

Some makers offer different levels of coverage and support
at different costs. Evaluate these based on your usage load
and up-time history. Also, carefully consider the state of the
art of the tool. The more modern it is, the more you will need
a maker's contract. Sad but true. More technologically advanced tools
lead to more opportunities for failures but also offers favorable
user interfaces and novel features.

gary g.


At 05:43 PM 10/26/2010, you wrote:
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From: nizets2-at-yahoo.com
Date: Wed, 27 Oct 2010 03:07:14 -0500
Subject: [Microscopy] viaWWW: Service insurance v. OEM service contracts

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

Hi Mark!

Be conscious that the insurance company will require you to make a yearly
maintenance service by the manufacturer and given that you will make no service
contract with them the maintenance will cost a lot.

Regards,

Stephane

 


----- Original Message ----
X-from: "mark.grimson-at-ttu.edu" {mark.grimson-at-ttu.edu}
To: nizets2-at-yahoo.com
Sent: Wed, October 27, 2010 2:46:37 AM

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Email: mark.grimson-at-ttu.edu
Name: Mark Grimson

Organization: Texas Tech University

Title-Subject: [Filtered] Service insurance v. OEM service contracts

Message: Hello, an insurance equipment company will be visiting us in
the near future in an attempt to talk us into getting a policy
through them rather than the manufacturer's service contract, which
we have had for 40 years with great success and working relationship.
I am sure the insurance might be ok for some equipment, but not SEMs
and TEMs.

  I am looking for any rationale I can use to help explain why the
Insurance route is not the way to go, and to stick with our
maintenance service contracts.  Anecdotes or any experiences with
insurance on em's will be greatly appreciated. Thanks, Mark

  Login Host: 129.118.38.156
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From: TindallR-at-missouri.edu
Date: Wed, 27 Oct 2010 08:59:01 -0500
Subject: [Microscopy] viaWWW: Service insurance v. OEM service contracts

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

Whenever this question arises, I feel duty-bound to chime in.

Stay with your manufacturer or a qualified independent service provider! Our two-year experience with insurance companies was disastrous. Virtually none of the promises they made were kept, such as "You will get the same level of service as you do now", or "You won't have to worry about paperwork". It was somewhat cheaper, but at what a cost.

It once took 9 months to arrange for a preventive maintenance visit on one of our scopes. We had to justify parts replacement and service calls above a certain dollar level. We had to wait in line behind the customers who had OEM service contracts until the companies could fit us in. Etc., etc., etc.

Once back on service contracts, all of these problems disappeared and we again received the same excellent level of service we had received before----no arguing over needed parts, no extended waits for service, no extra levels of bureaucracy to go through.

Stay with your OEMs or trusted independent. If yours are half as good as ours (JEOL, Hitachi and FEI), reward them with a little loyalty and it will pay off.

Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com


-----Original Message-----
X-from: mark.grimson-at-ttu.edu [mailto:mark.grimson-at-ttu.edu]
Sent: Tuesday, October 26, 2010 7:43 PM
To: Tindall, Randy D.

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Email: mark.grimson-at-ttu.edu
Name: Mark Grimson

Organization: Texas Tech University

Title-Subject: [Filtered] Service insurance v. OEM service contracts

Message: Hello, an insurance equipment company will be visiting us in
the near future in an attempt to talk us into getting a policy
through them rather than the manufacturer's service contract, which
we have had for 40 years with great success and working relationship.
I am sure the insurance might be ok for some equipment, but not SEMs
and TEMs.

I am looking for any rationale I can use to help explain why the
Insurance route is not the way to go, and to stick with our
maintenance service contracts. Anecdotes or any experiences with
insurance on em's will be greatly appreciated. Thanks, Mark

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From: klivi-at-jhu.edu
Date: Wed, 27 Oct 2010 09:02:32 -0500
Subject: [Microscopy] Re: viaWWW: Service insurance v. OEM service contracts

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

Dear Mark and others,

We currently have an insurance company (IC) as a middle man between us and our microscope company (MC). This is the way it works... We pay the IC to supply us with a service agreement at a price lower than what the MC charges. The agreement matches what the MC would supply (this is important) including preventative maintenance visits. How it is supposed to work is that when we have a service need, we call IC who calls MC who sends the engineer. How it actually works is that the MC will not honor the purchase orders of the IC, so we are forced to do it another way. We have to inform both IC and MC of our need, we then generate a PO# for MC, MC services the scope, we pay MC and IC reimburses us. I personally only have to do the first part, but other labs may have to do the PO generation and reimbursement paperwork.

So here are the pros and cons...
Pros - 1) We get a reduced price for a service contract. 2) The service comes from the same MC as before, so we are not reducing service quality. 3) We get to appease the Deans.

Cons - 1) More paper work for us. 2) Takes some time to generate PO# (depending upon your accounting system). 3) You are not top priority on the MC list (i.e., slower response time).

So, it really comes down to whether you think the increase in paperwork and slower response time is worth the money you save. Some have the resources, others do not (especially in these times). Interestingly, the MCs stand to gain by having people pay per diem and top dollar for parts. The microscopy community may benefit from competition on pricing.

I am not trying to convince anyone to go either way, but I thought it was important to properly describe this whole process and let managers decide upon facts.

Ciao for now,
Ken

On Oct 26, 2010, at 8:44 PM, mark.grimson-at-ttu.edu wrote:

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}
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} through them rather than the manufacturer's service contract, which
} we have had for 40 years with great success and working relationship.
} I am sure the insurance might be ok for some equipment, but not SEMs
} and TEMs.
}
} I am looking for any rationale I can use to help explain why the
} Insurance route is not the way to go, and to stick with our
} maintenance service contracts. Anecdotes or any experiences with
} insurance on em's will be greatly appreciated. Thanks, Mark
}
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|_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
Kenneth JT Livi, PhD
Director, The High-Resolution Analytical Electron Microbeam Facility
of the Integrated Imaging Center
Departments of Earth and Planetary Sciences and Biology
Olin Hall
3400 N Charles Street
Johns Hopkins University
Baltimore, Maryland 21218 USA
| | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| | | :|










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From: PhillipsT-at-missouri.edu
Date: Wed, 27 Oct 2010 10:52:54 -0500
Subject: [Microscopy] melanocytes

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

I realize this isn't strictly a microscopy question but more of a histology= question. I teach histology and therefore like to think I know a bit abou= t it but apparently not everything! I was looking at an unusual conjunctiv= al tissue and the blood vessels in one region are surrounded by melanocytes= . The overlying mucosa does not have melanoctyes. I know why the epidermis= has melanoctyes but can't rationalize why they would localize around blood= vessels. Anyone have insight? Thanks, Tom



Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/




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From: gradice-at-richmond.edu
Date: Wed, 27 Oct 2010 15:09:22 -0500
Subject: [Microscopy] re: melanocytes

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

I should point out this is a healthy avian tissue - it is not a melanocyte nevus.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: Derrick Horne [mailto:dhorne-at-interchange.ubc.ca]
Sent: Wednesday, October 27, 2010 11:21 AM
To: Phillips, Thomas E.

I don't know about mammals, but pigment cells in/on internal organs
are quite common in amphibians. Xenopus has a beautiful silver
pericardium, for example, because of the pigment cells there.

For a review, try: Moresco and Olivera, 2009, South American Journal
of Herpetology 4:1-8.
Available free online.

Searches for "extracutaneous pigmentary system" or extracutaneous
melanocytes" will also turn up leads.



Gary Radice
Department of Biology
University of Richmond
Richmond VA 23173
804-289-8107


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From: PhillipsT-at-missouri.edu
Date: Wed, 27 Oct 2010 18:23:23 -0500
Subject: [Microscopy] re: melanocytes

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

Gary's tip on using "extracutaneous melanocytes" was exactly what I needed. I had been using a combination of melanocyte AND blood vessels but that was pulling up all the pathology papers on melanocyte nevi. The thing I am looking at is in healthy tissue. Using this search term, I found a great paper (if you are interested- Plonka et al. (2009) Exp. Dermatol. 18:799-821 entitled "What are melanocytes doing all day?" In this paper, the author raises the question of the title and then 4 or 5 writers take turns trying to answer it in a page each followed by 6 different individuals making 1 page comments on the writers views. This seems like a neat concept.

Thanks for all the tips. Tom


-----Original Message-----
X-from: gradice-at-richmond.edu [mailto:gradice-at-richmond.edu]
Sent: Wednesday, October 27, 2010 3:10 PM
To: Phillips, Thomas E.

I don't know about mammals, but pigment cells in/on internal organs
are quite common in amphibians. Xenopus has a beautiful silver
pericardium, for example, because of the pigment cells there.

For a review, try: Moresco and Olivera, 2009, South American Journal
of Herpetology 4:1-8.
Available free online.

Searches for "extracutaneous pigmentary system" or extracutaneous
melanocytes" will also turn up leads.



Gary Radice
Department of Biology
University of Richmond
Richmond VA 23173
804-289-8107


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From: jrminter-at-rochester.rr.com
Date: Wed, 27 Oct 2010 18:38:08 -0500
Subject: [Microscopy] Re: viaWWW: Service insurance v. OEM service

Contents Retrieved from Microscopy Listserver Archives
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I wholeheartedly agree with Gary and the other posters who warned of the downside of insurance contracts versus OEM service. About 5 years ago one of the reputable insurance companies in the field convinced my management that we could save at least 15% by letting them underwrite the policies. The folks at the insurance company were helpful and processed the paperwork promptly. We discovered that the service on our FEG SEM and TEMs were the largest expenditure for our analytical department.

One major problem was that the vendor could not ship boards until a PO for the cost was received from the insurance company. This required a quote for the part and made it hard to get boards shipped out the same afternoon. Frequently the engineer has it narrowed down to a couple of boards. Under contract an engineer would get whatever parts might be needed for the next day sent out overnight and send back the unused ones. This was much more difficult under the insurance contract, because quotes would be needed for each part and the insurance company was understandably cost conscious.

As others have noted, contract customers take priority. This was an issue for us. The final straw was that it looked like we needed an HV tank for our 200 kV TEM. There was an exclusion for transformers in the insurance contract and my managers were not happy that we would be on the hook. This time it turned out to be a cable that was covered, but it could have been a major unplanned expense.

We finally put together a business justification to put our TEM and FEG-SEM back under vendor contract. Actually everybody was pleased with this. The insurance company is a good choice for instrumentation that does not need service frequently and does not use very expensive parts. We could easily choose alternate vendors when it made sense. We used this option with our microtomes. Another benefit of working with the insurance company was that we had put in an allotment for consumables. When we needed LaB6 cathodes and apertures the microscope vendor could ship and the insurance vendor would charge it to the right account - without us having to generate a purchase order. As I said, I am pleased with the insurance company for auxiliary equipment but think vendor service is best for high end SEMs and TEMs.

The last point I will make is that if you are considering an insurance provider is whether your microscope vendor will accept their POs or if you have to write a company PO and get reimbursed. At least one major microscope manufacturer will not accept POs from many insurance companies. They had too many problems collecting. The company we used had a history of prompt payment, so the microscope company agreed to continue to take their POs. People considering such options should have a frank discussion with the service manager for their microscope vendor.

Best Regards,
John Minter

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From: thoward-at-unm.edu
Date: Thu, 28 Oct 2010 12:01:02 -0500
Subject: [Microscopy] DyLight antibody conjugates?

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

I'm curious about the DyLight antibody conjugates - has
anyone tried them as secondaries for epifluorescence
and/or confocal? I've tried PubMed and searched the
archives for both this listserve and the confocal list &
haven't come up with anything useful as far as real life
experience with these fluors.

Thanks!

Tamara

***************************
Tamara Howard
Cell Biology & Physiology
UNM-HSC
Albuquerque, NM
***************************


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From: dhorne-at-interchange.ubc.ca
Date: Thu, 28 Oct 2010 20:52:11 -0500
Subject: [Microscopy] viaWWW: Image Drift Correction Software

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Email: dhorne-at-interchange.ubc.ca
Name: Derrick Horne

Organization: UBC BioImaging Facility

Title-Subject: [Filtered] Image Drift Correction Software in SEM and TEM

Message: Does anyone have experience with or knowledge of the current
availability of software that helps correct for image drift in SEM
and TEM images?

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From: gary-at-gaugler.com
Date: Thu, 28 Oct 2010 22:39:42 -0500
Subject: [Microscopy] Re: viaWWW: Image Drift Correction Software

Contents Retrieved from Microscopy Listserver Archives
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what SEM and what TEM and what application??

gary g.



At 06:53 PM 10/28/2010, you wrote:



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From: dhorne-at-interchange.ubc.ca
Date: Fri, 29 Oct 2010 13:23:54 -0500
Subject: [Microscopy] Re: viaWWW: Image Drift Correction Software

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

Hi Gary,

That's a valid question.

The immediate need is for SE/BSE imaging on a Hitachi S4700 FESEM, but
we also run an H7600 TEM, a S2600 VP-SEM, and a Tecnai G2 20 Twin. The
motor drive on the Tecnai does a more than adequate job of maintaining
the stage position during tomography acquisitions, and the built in
software is fantastic at aligning the data sets. The FESEM is a little
trickier as there is no feedback to the stage from the image.

We're not all that interested in pairing up image drift in EDS
applications at this point as we do not currently run that on our
FESEM. Essentially, what we're hoping to do is take full advantage of
the resolution of the FESEM in situations where our users cannot stop
image drift by adjusting the SEM/sample setup, and salvage data by
"reconstructing" the image. The source of the drift is variable, and
it is apparent that sometimes it is mechanical (e.g. a poorly isolated
compressor), and sometimes it appears to be electrostatic drift.

We spend much time working with our users to ensure they have the best
preparation possible, but in the case of sub-optimal preparation we
would like a plan B.

I should mention that we are mainly, but not exclusively, imaging
biological samples at 100kx or greater, and the drift we're talking
about is in the range of a few pixels in a 1280x960 image over an 80s
line capture. The features of interest that become obscured are in the
5-20nm range. When we are imaging materials samples (e.g. quantum dots
or nanotubes) we could be attempting to image at magnifications .250kx.

I hope this is a clear explanation of what is happening, but I would
be more than happy to clarify anything I've stated.

Thanks,

Derrick



On 28-Oct-10, at 8:39 PM, Gary Gaugler wrote:

} what SEM and what TEM and what application??
}
} gary g.
}
}
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} At 06:53 PM 10/28/2010, you wrote:
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} } Title-Subject: [Filtered] Image Drift Correction Software in SEM
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From: johnf-at-geology.wisc.edu
Date: Sat, 30 Oct 2010 12:28:48 -0500
Subject: [Microscopy] RE: problem with Hitachi S3400 W filaments

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

Thanks to everyone who offered suggestions.

On Oct 22 we removed the "new" Wehnalt which had been in operation a
couple of weeks. There was a purple-blue tint around the opening.
Installing a "totally new" Wehnalt and then examining the slightly
used but discolored Wehnalt, it was not obvious (even dropping E0 to
5 kV) whether any elements present beside W in EDS spectrum. However,
upon close inspection of the inner walls of the aperture, there was a
7 micron thick band around the whole opening, which by EDS indication
was a tungsten oxide. Then very close inspection of a hard to see
portion of the ceramic in the gun showed some light brownish
discoloration.

Many people may not be aware of (and I certainly wasn't): there is NO
high vacuum gauge in the Hitachi S3400! ALL vacuum readings and
decisions of the automation are based upon readings of Pirani
(thermocouple) gauges. Ergo, there was nothing in the operation of
the system to indicate a large leak. (Poor design in my book).

Given the tungsten oxide plus the suggestions by several folks on
this list, it was obvious to me that there had to be a lead, which
would explain the tungsten oxide coating as well as the galling of 2
Wehnalt caps.

Yesterday our Hitachi FSE installed an ionization gauge on the system
(above the turbo pump, below the gate valve to the chamber, in line
with the pumping line on the gun. First reading in high vac mode was
1x10-3 torr, whereas in VP mode (chamber closed to direct reading of
gauge) it was 1x10-4 torr. The published "ultimate" vacuum for the
high vac mode is 1x10-5 torr.

--} A big leak in the chamber it would seem!

After 10 minutes of so of searching, the leak was found (had to use a
flashlight)... a virtually invisible small stiff hair/fiber laying
across the chamber door o-ring. Inspection of the opposite surface
showed a small amount of oil surrounding that area. Pumping back
down, we achieve 10-4 torr range immediately. Pumping now for 17
hours has us at 2.2x10-5 torr.

Lesson 1: If your SEM doesn't have a high vacuum gauge, you risk the
problems we went thru. Problems with short filament life will be
assumed to be operator's problems with oversaturation, when they
could be a vacuum leak.
Lesson 2: If at time of purchase I knew that there was no high vac
gauge, I would have required that to be installed, and even if it
cost another $1500 or so, it would definitely be worth it.

We will be installing an ionization gauge on our SEM. Hitachi is
allowing us to borrow theirs until we get our own installed. And they
did provide us with a replacement Wehnalt and a box of filaments at
no cost.

John Fournelle

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--
========================================================
John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480
Cameron Electron Microprobe Lab lab: (608) 265-4798
Department of Geoscience fax: (608) 262-0693
University of Wisconsin home: (608) 274-2245
1215 West Dayton St. email: johnf-at-geology.wisc.edu
Madison, WI 53706 amateur radio: WA3BTA
Personal http://www.geology.wisc.edu/~johnf/
Probe lab http://www.geology.wisc.edu/~johnf/sx51.html
Probe Sign Up Calender:
http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi

"The first rule of all intelligent tinkering is to save every cog and
wheel." -- Aldo Leopold

"For a successful technology, reality must take precedence over
public relations, for Nature cannot be fooled." -- Richard P.
Feynman

==============================Original Headers==============================
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15, 25 -- Subject: [Microscopy] RE: problem with Hitachi S3400 W filaments
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From: dsherman-at-purdue.edu
Date: Sat, 30 Oct 2010 13:02:21 -0500
Subject: [Microscopy] TEM camera recommendations

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


Hi all,

We are considering adding a 2K camera (side-mount) to a TEM for primarily
live image previewing of samples, including low light situations. I would
appreciate hearing from those of you with recommendations/comments as to
camera models, reliability, ease of software use in a multi-user
environment, and details of pre- and post-install interactions with
vendors. Options on cooled verses non-cooled for this resolution camera
would be appreciated.

Please no vendor response at this time.
It would be best to post responses off-line.

Thanks,
Debby
-
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy/



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From: bozzola-at-siu.edu
Date: Sat, 30 Oct 2010 13:34:59 -0500
Subject: [Microscopy] problem with Hitachi S3400 W filaments

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Excellent, and significant observations! I believe it's important (for
monitoring purposes) that EMs be equipped with vacuum gauges capable
of reading in the high vacuum range. Whether this is a cold cathode or
heated cathode ionization gauge is up to the end user.

The most critical area is obviously the gun, but some older
microscopes cannot readily be retrofitted in-house in the gun area
(unless you have techs who can tie into vacuum lines). Most of our
EMs, even a 28 y/o SEM, have ionization gauges to read vacuum levels.
Many times, the vacuum readings have indicated problems (such as a
damaged O-ring -- or a fiber across one) and we took corrective action
before losing a filament. The key is that you have to monitor the
readings to see if there is a loss of vacuum. Don't just rely on the
automatic valving system.

A word of caution if you add an ionization gauge to the specimen
chamber area. This is the easiest place to locate a gauge due to the
many ports that are easily modified to accept a vacuum probe. However,
the light generated during the ionization process may blind (or
possibly damage) the photomultiplier (PM) in your SE detector. In our
old SEM, we have a circuit that shuts off the kV and PM when the
vacuum gauge is activated, hence preventing damage.

If you are fortunate enough to be shopping for a new EM, then insist
that HV gauges be installed in the gun area. The several thousand
dollars of added cost will save you many filaments and headaches over
the years.

John Bozzola
IMAGE Center
Southern Illinois University
Carbondle, IL

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From: TindallR-at-missouri.edu
Date: Mon, 1 Nov 2010 11:12:03 -0500
Subject: [Microscopy] Cryoultramicrotomy: Ion generator---or not?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Collective,

Our Diatome Staticline II ion generator for cryoultramicrotomy seems not be generating ions. We had a mangled cable repaired by our crack fine instrument repair shop, but this did not cure the ailment, so we may need to purchase a new one. The tip seems to be in good condition.

One question, though, is how do you tell if an ion beam is being generated, short of actually firing up the cryo-UM? I've never noticed that it has any visible effect on the nitrogen vapor as it goes through it, otherwise we could just test it on a little bowl of LN2.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




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From: spurgeon-at-drexel.edu
Date: Mon, 1 Nov 2010 11:44:35 -0500
Subject: [Microscopy] FIB / Atom Probe - Commercially available Si microtip array?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hello everyone,

I am looking to prepare a large number of samples for atom probe
tomography using a FIB lift out technique. I was reading a paper by
Thompson et al. who describe the use of a MEMS-fabricated array of Si
microtips to mount their lift out sections prior to annular milling.
The image given in their paper can be found here:
http://i56.tinypic.com/2e188d3.jpg (Sorry, scale bar is missing)

My question is: are there any commercial vendors that sell such
arrays? If not, how difficult would it be to prepare something like
this via a lithography technique? Any suggestions would be greatly
appreciated. Thanks!

The reference for the paper I mentioned is: Thompson, K. et al. "In
situ site-specific specimen preparation for atom probe tomography."
Ultramicroscopy. 107:131-139 (2007).
doi:10.1016/j.ultramic.2006.06.008

--
Steven Spurgeon
Dynamic Characterization Group
Department of Materials Science & Engineering
Drexel University

Cell: 719.330.0441
Office: Bossone 102
Email: spurgeon-at-drexel.edu
Web: http://www.materials.drexel.edu/dcg/

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From: lcgould-at-med.cornell.edu
Date: Mon, 1 Nov 2010 13:14:21 -0500
Subject: [Microscopy] Leica Ultracut S parts

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi All-
My Leica Ultracut S fried its lamp housing last week. I've been told that its the oscillator board. I've also been told that there are no parts out there of any kind for the "S" since it had a short production run and is no longer supported by Leica. Does anyone out there have parts? I would even be willing to buy the whole microscope carrier assembly.
thanks

Lee


Lee Cohen-Gould, M.S., C.E.M.T.
Sr. Staff Associate in Biochemistry and
Cell & Developmental Biology
Director, Electron Microscopy & Histology
and Optical Microscopy Core Facilities
Weill Cornell Medical College

voice (212)746-6146
fax (212)746-8175
http://www.med.cornell.edu/research/rea_sup/
http://www.cornellcelldevbiology.org
http://www.cornellbiochem.org


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From: pmachado-at-igc.gulbenkian.pt
Date: Mon, 1 Nov 2010 13:40:46 -0500
Subject: [Microscopy] Re: Leica Ultracut S parts

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Lee,

We had the same problem here (I'm guessing it's the same board, one
located above the lamps) almost 6 months ago with our ultracut s. Leica
could not solve the problem (for reasons that are not valid in my
opinion).
The maintenance people here manage to build an external power source for
the lamps (not connected to the microtome interface).

X-from what I learned any person qualified on electrical/lighting
maintenance can solve your problem. Our board took a while to arrive but
now all is well (5 months total time, Leica saga took 3 ou of 5).

Oh, another important detail is that those kind of lamp are rare now, so
you should have a stock at hand.

I hope this can help you.

Good luck,
Pedro Machado
Instituto Gulbenkian de Ciencia
PT

}
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} Hi All-
} My Leica Ultracut S fried its lamp housing last week. I've been told that
} its the oscillator board. I've also been told that there are no parts out
} there of any kind for the "S" since it had a short production run and is
} no longer supported by Leica. Does anyone out there have parts? I would
} even be willing to buy the whole microscope carrier assembly.
} thanks
}
} Lee
}
}
} Lee Cohen-Gould, M.S., C.E.M.T.
} Sr. Staff Associate in Biochemistry and
} Cell & Developmental Biology
} Director, Electron Microscopy & Histology
} and Optical Microscopy Core Facilities
} Weill Cornell Medical College
}
} voice (212)746-6146
} fax (212)746-8175
} http://www.med.cornell.edu/research/rea_sup/
} http://www.cornellcelldevbiology.org
} http://www.cornellbiochem.org
}
}
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From: bozzola-at-siu.edu
Date: Mon, 1 Nov 2010 14:14:12 -0500
Subject: [Microscopy] Re: Cryoultramicrotomy: Ion generator---or not?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Randy,

Easiest way would be to generate static and see if it is neutralized.

For example: grids in a petri dish that is slid across a plastic
surface will cause the grids to fly to the top of the dish. The ion
gun should neutralize the charge, causing the grids to drop back onto
the filter paper in the plate.

Or, you could rub a balloon against a cat and see if it causes the
balloon to drop off.

John Bozzola

--
John J. Bozzola, Ph.D., Professor & Director of IMAGE
Integrated Microscopy & Graphics Expertise
Southern Illinois University
750 Communications Drive
Carbondale, IL  62901
Phone: 618-453-3730


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From: speransv-at-mail.nih.gov
Date: Mon, 1 Nov 2010 14:20:02 -0500
Subject: [Microscopy] Cryoultramicrotomy: Ion generator---or not?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Ours came with this little thing called Multi Check testing device. It lights up when facing a working ionizer tip. Maybe you have it too somewhere?
If not, I would contact Al Coritz, now at Diatome US. Perhaps he can suggest other ways to test.

Vlad
________________________________________________
Vlad Speransky, Staff Scientist
Biomedical Engineering and Physical Science Shared Resource
National Institute of Biomedical Imaging and Bioengineering, NIH
13 South Dr, Rm. 3N17 MSC 5766
Bethesda, MD 20892
301 496-3989
vladislav_speransky-at-nih.gov
http://www.nibib.nih.gov/Research/Intramural/SharedResource/Speransky


________________________________________
X-from: TindallR-at-missouri.edu [TindallR-at-missouri.edu]
Sent: Monday, November 01, 2010 12:23 PM
To: Speransky, Vlad (NIH/NIBIB) [E]

Dear Collective,

Our Diatome Staticline II ion generator for cryoultramicrotomy seems not be generating ions. We had a mangled cable repaired by our crack fine instrument repair shop, but this did not cure the ailment, so we may need to purchase a new one. The tip seems to be in good condition.

One question, though, is how do you tell if an ion beam is being generated, short of actually firing up the cryo-UM? I've never noticed that it has any visible effect on the nitrogen vapor as it goes through it, otherwise we could just test it on a little bowl of LN2.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




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From: TindallR-at-missouri.edu
Date: Mon, 1 Nov 2010 14:26:22 -0500
Subject: [Microscopy] Ionizer----oops

Contents Retrieved from Microscopy Listserver Archives
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Thanks to those who replied to my question about how to detect ions. Turns out we have this thing called a Multi Check that is an...ahem....ion detector for the unit. Soon as I get the battery replaced, we are good to go. Sheeesh, Tindall......

Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com




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From: allan.mitchell-at-stonebow.otago.ac.nz
Date: Mon, 1 Nov 2010 14:31:52 -0500
Subject: [Microscopy] Re: Cryoultramicrotomy: Ion generator---or not?

Contents Retrieved from Microscopy Listserver Archives
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Hi Randy

Our staticline ioniser came with a little hand held device you place
in front of the ioniser when switched on which checks ion discharge.
A green LED comes on if there are ions being generated. The device
was called a MultiCheck. (Turn on ioniser (at RT) move nose of tester
towards ioniser, at about 4 cm the red light should glow green).

I guess another check to see if it is functioning would be to turn it
onto full power when cryosectioning and see if the cryosections fly
away. Do they stop flying away as the power is reduced?

I did a cryo-course in Utrecht a couple of years back and a couple of
things came up at this course about ionisers that I did not know
about, and was not told about when we purchased it. These notes are
below:
1. Ioniser not working - clean nozzle parts with brush to remove
ice. Ice build up will reduce effect.

2. Tips erode over time and become covered in contamination (they
blacken). Can be cleaned with sandpaper and gentle rubbing.
3. Tips erode fast if taken out of cryochamber with transformer
still on. Can be stuffed with an 1 hour.

4. Otherwise should last at least 2 years before electrode requires
replacement (so they do need replacement).

There was also some discussion about the voltage regulator playing up
sometimes and needing replacement. I recall something about being
careful to get the correct voltage (i think) for replacement)

Hope this helps

Allan


On 2/11/2010, at 5:21 AM, {TindallR-at-missouri.edu}
{TindallR-at-missouri.edu} wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear Collective,
}
} Our Diatome Staticline II ion generator for cryoultramicrotomy seems
} not be generating ions. We had a mangled cable repaired by our
} crack fine instrument repair shop, but this did not cure the
} ailment, so we may need to purchase a new one. The tip seems to be
} in good condition.
}
} One question, though, is how do you tell if an ion beam is being
} generated, short of actually firing up the cryo-UM? I've never
} noticed that it has any visible effect on the nitrogen vapor as it
} goes through it, otherwise we could just test it on a little bowl of
} LN2.
}
} Cheers,
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
} Tel: (573) 882-8304
} Fax: (573) 884-2227
} Email: tindallr-at-missouri.edu
} Web: http://www.emc.missouri.edu
} On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
} Sons of Norway: http://www.sofn.com
}
}

Allan Mitchell
Microscopy Otago - Electron Microscopy
c/- Department of Anatomy and Structural Biology
Otago School of Medical Sciences
P.O. Box 913
Dunedin
New Zealand

Phone (03) 479 5642 or 479 7301
Fax (03) 479 5086 or 479 7254

EM Centre: http://ocem.otago.ac.nz/
Confocal Centre: http://occm.otago.ac.nz/
NZ Microscopy Society: http://microscopynz.co.nz/




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From: joelsheffield-at-gmail.com
Date: Mon, 1 Nov 2010 16:25:13 -0500
Subject: [Microscopy] re: Aldehyde and buffer concentration

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Let me add one point to this discussion. It is clear that during the
process of fixation a series of changes occur to the membranes
of cells, as their proteins become cross linked, precipitated, etc. Thus, a
simple calculation of tonicity may not be relevant since the permeability of
the cells changes during the process. During my thesis work, in the dark
ages, I had the good fortune to be working with populations of dissociated
retina cells of many sizes in suspension, and a Coulter Counter which
allowed analysis of cell size. These days, you could do it with a FACS
machine. At any rate, I experimented with various buffer concentrations and
fixatives so as to get a size distribution of fixed cells that was similar
to that of the unfixed cells. My final fixative mixture was 2.5%
glutaraldehyde in 0.08M Sodium Phosphate. Your results may vary. (Sheffield
and Moscona, 1970, Develop. Biol *23*:36-31, and Sheffield, J. Morphol, *132
*:245-264, 1970.

Joel Sheffield


On Wed, Oct 20, 2010 at 4:06 PM, |--leunissen-at-aurion.nl--| wrote:



------------------------------
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--| ----------------------------------------------
--| The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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--|
--| Dear list
--|
--| I have never quite understood the relevance of the concentration of the
--| buffer components in glutaraldehyde or formaldehyde fixatives.
--| No doubt there is an effect of the buffer, this has been amply illustrated.
--| But if one considers the contribution of the aldehydes to the molarity of
--| the fixative solution, it seems something else than tonicity is at play.
--|
--| Monomeric glutaraldehyde has a Mwt of 100. A 4% solution corresponds to
--| 0.4M.
--|
--| The situation is even more strange for formaldehyde with a Mwt of 30. A 4%
--| solution being 1.33M.
--|
--| Clearly at the very onset of fixation the situation is that the fixative is
--| highly hypertonic.
--| Perhaps it is rather the buffer capacity than the osmolarity that is
--| important?
--|
--| Always keen to learn..
--|
--| Jan Leunissen
--|
--| Aurion
--|



--
Joel B. Sheffield, Ph.D.
Biology Department, Temple University
1900 North 12th Street
Philadelphia, PA 19122
jbs-at-temple.edu
(215) 204 8839, fax (215) 204 0486
http://astro.temple.edu/~jbs

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From: bill.mcmanus-at-usu.edu
Date: Mon, 1 Nov 2010 17:12:42 -0500
Subject: [Microscopy] Blown Turbo pump

Contents Retrieved from Microscopy Listserver Archives
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Hello all:

After not being used for some time, when started the Varian turbo V200
in my IBS coater, the pump went into a full scream. By time I got it
turned off there were metal fragments on the little magnet in the oil
sump. Does anyone out there happen to have an extra laying around, or
one that is compatible? I could pay a reasonable price for a working
pump.

Thanks for any help

Bill McManus
Research Scientist
Western Dairy Center
Utah State University
bill.mcmanus-at-usu.edu


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From: swtkeller-at-yahoo.com
Date: Mon, 1 Nov 2010 17:27:55 -0500
Subject: [Microscopy] viaWWW: TEM-looking for a compatable computer or card/cable for

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Email: swtkeller-at-yahoo.com
Name: Sandra keller

Organization: TA-SICCO

Title-Subject: [Filtered] TEM-looking for a compatable computer or
card/cable for Gatan 694 camera

Message: Hi All:
I am trying to resurrect an old Gatan 694 camera. I am told I should look for:
1) a Gatan DMA Mac NuBus card
2) a Mac DMA cable that goes from the computer to the SSC camera controller
3)a computer that runs MacOS 9.0.2 to support the camera,

If none of these components are available, I would consider a
complete functioning Gatan 694 camera system with all the
components). Please feel free to contact me offline with any offers...

BR,
Sandra

Login Host: 71.91.112.63
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From: Elliott-at-arizona.edu
Date: Mon, 1 Nov 2010 19:16:59 -0500
Subject: [Microscopy] re: Aldehyde and buffer concentration

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I like the estimate of the effective osmolality given in Maunsbach and
Afzelius "Biomedical Electron Microscopy"

Osm (eff) = Osm (buffer) + 0.3 x Osm (glut) + 0.1 x Osm (form)

It's not perfect, but it is a very good starting point when I am
developing a fixative.

David

------------------------------
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--| ----------------------------------------------
--| The Microscopy ListServer -- CoSponsor: The Microscopy Society of
America
--| To Subscribe/Unsubscribe --
--| http://www.microscopy.com/MicroscopyListserver
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--| http://www.microscopy.com/MicroscopyListserver/FAQ.html
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----------------------------------------------------------------------------
--|
--| Dear list
--|
--| I have never quite understood the relevance of the concentration
of the
--| buffer components in glutaraldehyde or formaldehyde fixatives.
--| No doubt there is an effect of the buffer, this has been amply
illustrated.
--| But if one considers the contribution of the aldehydes to the
molarity of
--| the fixative solution, it seems something else than tonicity is at
play.
--|
--| Monomeric glutaraldehyde has a Mwt of 100. A 4% solution
corresponds to
--| 0.4M.
--|
--| The situation is even more strange for formaldehyde with a Mwt of
30. A 4%
--| solution being 1.33M.
--|
--| Clearly at the very onset of fixation the situation is that the
fixative is
--| highly hypertonic.
--| Perhaps it is rather the buffer capacity than the osmolarity that is
--| important?
--|
--| Always keen to learn..
--|
--| Jan Leunissen
--|
--| Aurion
--|



--
Joel B. Sheffield, Ph.D.
Biology Department, Temple University
1900 North 12th Street
Philadelphia, PA 19122
jbs-at-temple.edu
(215) 204 8839, fax (215) 204 0486
http://astro.temple.edu/~jbs

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10, 30 -- Subject: re: Aldehyde and buffer concentration
10, 30 -- From: Joel Sheffield {joelsheffield-at-gmail.com}
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From: jflaci-at-ms.sapientia.ro
Date: Tue, 2 Nov 2010 03:49:10 -0500
Subject: [Microscopy] Re: viaWWW: TEM-looking for a compatable computer or

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi there!

I'm using the same camera on our JEOL 100U.

The Mac we are using is a Quadra 700, but this:

http://cgi.ebay.com/Macintosh-Quadra-800-great-condition-/170558285904?pt=LH_DefaultDomain_0&hash=item27b610e850

would be appropiate for You.

The main problem is the DMA card, as the cable is simple to make, is a
pin-to-pin cable, any electronist could make that for You.
I know this for sure, as a colleague of mine made ours, as we missed it also.

Laci

PS: do You have that vacuum chamber for the camera which fixes it to
the microscope?

==============================Original Headers==============================
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From: CGoldsmith-at-cdc.gov
Date: Tue, 2 Nov 2010 08:13:01 -0500
Subject: [Microscopy] viaWWW: PhD position with the Health Protection Agency in London

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Email: CGoldsmith-at-cdc.gov
Name: Cynthia Goldsmith

Organization: Centers for Disease Control and Prevention (CDC)

Title-Subject: [Filtered] PhD position with the
Health Protection Agency in London

Message: Job title: Lead Electron Microscopist
Division: Centre for Infections, Colindale
Department: Virus Reference Department (VRD)
Accountable to: Deputy Director, VRD
Salary: £43,202 - £50,972 per annum
Grade: AfC Band 8a
Location: Colindale, NW London
Hours: 37.5 per week
Ref: 919-CFI-MK-6983877

The Health Protection Agency (HPA) is an
independent body that protects the health and
well-being of everyone in England and Wales. The
Agency plays a critical role in protecting people
from infectious diseases and in preventing harm
when hazards involving chemicals, poisons or
radiation occur. We also prepare for new and
emerging threats, such as a bio-terrorist attack
or virulent new strains of pathogens.

The Centre for Infections at Colindale (CfI), NW
London is an internationally renowned centre of
excellence for expertise, reference microbiology,
research, surveillance and epidemiology in
relation to human health, working directly and
indirectly at the local, regional, national and
international level. The modern purpose built
centre is well equipped and provides excellent
Library, Occupational Health and Refectory
facilities. There is also a Nursery and Gym on
site.

An exciting opportunity has opened up for an
experienced post-doctoral electron microscopist.
The Electron Microscopy facility is currently
being completely refurbished, including the
installation of a new transmission electron
microsope. While it is situated in the Virus
Reference Department, it offers a service to all
departments within CfI, has close working
relationships with the other arms of HPA
Microbiology, and provides expert and specialist
services to the NHS, academic partners, and
others. The facility is a critical component of
HPAís capability to respond both to emergencies,
emerging infections and the investigation of
diseases of suspected but unknown infectious
Êtiology. It also contributes significantly to
R&D activities, including the development of
novel diagnostic tools. The newly refurbished
facility will incorporate a JEOL 1400TEM fitted
with an AMT digital camera and tomography,
together with an off-microscope workstation, and
will be equipped for both negative staining and
thin sectioning.

Ideally, the successful candidate will have a PhD
(or equivalent experience) with substantial
relevant specialist knowledge and practical
experience in EM and associated techniques.
While service delivery will be a priority we are
looking for an individual with the energy and
vision to take and develop opportunities to make
optimal use of the facilities, expertise and
scope of the health protection activities in the
HPA and to develop their own research projects.
The National Institute for Biological Standards &
Control recently joined HPA, providing access to
additional imaging facilities sited there. These
include SEM, cryo-TEM, cryo-SEM, confocal
microscopy and live cell imaging, broadening
experience and opportunity, as well as plans for
complementary and synergistic working, while
further strengthening imaging capabilities within
the HPA.

For an informal discussion please contact Prof
John Parry on ext 6208 or Prof David Brown on ext
6018.

Closing date: 15 November 2010

For more information about the HPA see the HPA website www.hpa.org.uk/careers

The Health Protection Agency promotes diversity
in the workplace and is an equal opportunities
employer.


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From: kenconverse-at-qualityimages.biz
Date: Tue, 2 Nov 2010 08:21:31 -0500
Subject: [Microscopy] Blown Turbo pump

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Bill,
Have you looked at Duniway Stockroom?
www.duniway.com
or more specifically
www.duniway.com/images/pdf/pg/p-95-turbo-pumps-rebuilt.pdf

No financial interest, just a happy customer.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: bill.mcmanus-at-usu.edu [mailto:bill.mcmanus-at-usu.edu]
Sent: Monday, November 01, 2010 6:14 PM
To: kenconverse-at-qualityimages.biz

Hello all:

After not being used for some time, when started the Varian turbo V200
in my IBS coater, the pump went into a full scream. By time I got it
turned off there were metal fragments on the little magnet in the oil
sump. Does anyone out there happen to have an extra laying around, or
one that is compatible? I could pay a reasonable price for a working
pump.

Thanks for any help

Bill McManus
Research Scientist
Western Dairy Center
Utah State University
bill.mcmanus-at-usu.edu


==============================Original Headers==============================
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From: brad-at-nanounity.com
Date: Tue, 2 Nov 2010 15:11:56 -0500
Subject: [Microscopy] viaWWW: Northern California Society for Microscopy

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Email: brad-at-nanounity.com
Name: Brad Rangell

Organization: Nanounity

Title-Subject: [Filtered] Northern California Society for Microscopy

Message: Dear Subscribers,

This message is regarding local affiliate societies. There is a group
of interested members in the process of reinstating the Northern
California Society for Microscopy (NCSM). Currently, we have about 60
individuals and 3 corporate sponsors that have committed to
membership. We are in the membership drive phase and want to
encourage other interested people to join. If you are in the Northern
California region and would like to sign up you can send an email to
the contacts below. In the coming few weeks, a date will be picked
for an inaugural meeting.

Membership contacts:

Brad Rangell
brad-at-nanounity.com

Steven Samuelsson
Steven.samuelsson-at-sri.com


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From: rbeavers-at-mail.smu.edu
Date: Tue, 2 Nov 2010 15:26:08 -0500
Subject: [Microscopy] Shipping Leo 906E TEM

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Do any of you have dimensions and weight requirements for shipping a Leo 906E TEM

Roy Beavers

Southern Methodist University
Department of Earth Sciences
P.O. Box 750395
Dallas, TX 75275
Voice: 214-768-2756
Fax: 214-768-2701
Email: rbeavers-at-smu.edu



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From: alice.dohnalkova-at-pnl.gov
Date: Tue, 2 Nov 2010 16:16:24 -0500
Subject: [Microscopy] viaWWW: stabilizing biofilm?

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Email: alice.dohnalkova-at-pnl.gov
Name: Alice Dohnalkova

Organization: Pacific Northwest National Laboratory

Title-Subject: [Filtered] stabilizing biofilm?

Message: Dear List,

Could you please recommend a method for stabilizing / immobilization
of a natural biofilm (typical thickness 5-10 mm) for EM processing? I
was told this biofilm doesn't hold together too well, and the
sampling site is pretty remote, so there is a good chance it would
get damaged by the time it will reach me.
The sampling team will have 2.5% glut available, but no means of
preparing / heating e.g. agarose or gelatin (that would be the idea
of putting a thin layer on top of a biofilm).

I'd really appreciate any suggestion.
Thanks!
Alice.

Alice Dohnalkova
Sr. Research Scientist
ENVIRONMENTAL MOLECULAR SCIENCES LABORATORY
Pacific Northwest National Laboratory
902 Battelle Boulevard
P.O. Box 999, MSIN K8-93
Richland, WA 99352 USA
Tel: 509-371-6515
Fax: 509-371-6242
Alice.dohnalkova-at-pnl.gov
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From: jtwilley-at-sprynet.com
Date: Tue, 2 Nov 2010 22:57:43 -0500
Subject: [Microscopy] Identification of fossilized tissue

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I would like to ask for the assistance of anyone with a background in biology who might be able to differentiate plant fiber from leather or animal tissue preserved in an iron oxide concretion. SEM views of a cross section show well-preserved cellular microstructures of some organic precursor pseudomorphically replaced by iron oxide corrosion products. I can forward photos off-list.

John Twilley

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From: marie.cantino-at-uconn.edu
Date: Wed, 3 Nov 2010 15:08:01 -0500
Subject: [Microscopy] EDS detection of Boron

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Can anyone give me a typical detection limit for boron in an organic
matrix using a thin window EDS detector?

Thanks!

Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369


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From: mike.flaws-at-canterbury.ac.nz
Date: Wed, 3 Nov 2010 16:10:48 -0500
Subject: [Microscopy] Hitachi H600 available

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Hi,

We have a 1979 Hitachi H600 TEM available complete or as parts. The
microscope is in reasonable condition although the rotary pumps could do
with a service.

Mike Flaws
Technical Services Manager
Department of Mechanical Engineering
University of Canterbury
Private Bag 4800
Christchurch
New Zealand

Telephone: +64-3-366 7001 Ext 7354
Facsimile: +64-3-364 2078
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From: wesaia-at-iastate.edu
Date: Wed, 3 Nov 2010 17:22:51 -0500
Subject: [Microscopy] Polaron E5100 sputter coater available

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We have taken delivery on a new, high-resolution sputter coater. That means we need to move our old Polaron E5100 out of the lab. (That's the model with the annular target.)

Is there anyone out there interested in it that we should go through the steps of listing as excess equipment? It still works but pumps down slowly.

We have one gold and one silver target with a lot of life in them (guessing upwards of 50%). We could include those or send them back for credit to the folks that have been providing us with our gold targets of late.

We have a direct-drive rotary pump on the unit now, but we intend to keep it. We do have a nice, belt-drive, 100-lpm rotary pump that we have kept around from an old microprobe that would probably do well on the coater.

Is anyone interested?
Please reply directly.

Warren Straszheim
515-294-8187


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From: mjbehr-at-dow.com
Date: Wed, 3 Nov 2010 20:29:33 -0500
Subject: [Microscopy] viaWWW: Microanalysis standard-revised

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Email: mjbehr-at-dow.com
Name: Michael Behr

Title-Subject: [Filtered] Microanalysis standard-revised

Message: I am seeking recommendation for replacement of NIST srm
2063a thin film microanalysis standard. Is there a newer version of
this available, or is someone willing to sell lend/sell a 2063a
standard to me? (I had the numbers backwards in previous
posting).Thank you!

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From: gary-at-gaugler.com
Date: Wed, 3 Nov 2010 20:40:34 -0500
Subject: [Microscopy] Re: EDS detection of Boron

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I'm not sure what you mean here or are seeking.
What do you define as the "detection limit?"

A good Si(Li) detector will barely detect
B at Ka=183eV but a modern SDD will do much
better. I work with non-bio specimens mostly
for EDS but for low Z work, low KV works well.

Perhaps you mean best resolution? That is product
specific and also condition specific. Tell us
more and we might be able to help to a more useful
level.

gary g.

Sigh...here come all of the out of office replies.


At 01:09 PM 11/3/2010, you wrote:



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From: eschumacher-at-mccrone.com
Date: Thu, 4 Nov 2010 07:23:56 -0500
Subject: [Microscopy] Emergency Plan: Restoring Analytical Services

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Greetings Fellow Microscopists,

We've had some discussions here recently on how we might quickly regain at least some capabilities should we suffer a natural disaster or similar event that disabled some or all of our equipment, or made our laboratory space temporarily unusable. We wondered if other labs have considered these scenarios, formulated action plans, or have possibly gone through such an event and have experiences to share. Your input is greatly appreciated; I'll plan to post a summary of responses.

Best regards,

Elaine

*********************************************************************
Elaine F.Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com



*********************************************************************
This message and any attachments are solely for the
intended recipient. If you are not the intended recipient,
disclosure, copying, use or distribution of the information
included in this message is prohibited.
*********************************************************************


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From: TindallR-at-missouri.edu
Date: Thu, 4 Nov 2010 10:05:43 -0500
Subject: [Microscopy] So long and Thanks

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Collective,

I'm getting ready to take a break from the world of Electron Microscopy and just wanted to thank all the Denizens of the List who have helped rescue my hindquarters over the years. Seemed like no matter how deep the hole I was in, someone out there had the right rope to pull me out and the generosity to throw it to me.

This is a really great resource thanks to all of you, and, of course, our Friendly Neighborhood Sysop Nestor, who keeps it running smoothly. It has been a great pleasure to be part of it.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: maryflet-at-interchange.ubc.ca
Date: Thu, 4 Nov 2010 11:02:35 -0500
Subject: [Microscopy] position available at UBC, Vancouver, Canada

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Listserver,

Due to my upcoming retirement, my position will be opening up to someone who
likes to work with electron microscopes.

Position Available:

Electron Microscopy Technician



The Department of Materials Engineering at the University of British
Columbia is currently seeking a technician to oversee their electron
microscopy laboratory. The primary functions of this position are to
maintain the electron microscopes (SEM and TEM), to train graduate students
in the use of analytical electron microscopes and to facilitate the
undergraduate teaching that makes use of the electron microscopy facilities.
The candidate should have previous experience in the use and maintenance of
electron microscopes. Previous experience as a technician in an academic
electron microscopy laboratory would be an asset. For more information
please see: "www.mtrl.ubc.ca/diverse/EMpositionMTRLUBC.pdf

To Apply: Please email your CV and cover letter to Mrs. Fiona Webster:
fiona.webster-at-apsc.ubc.ca



I hope we get someone soon enough for me to train them.



Regards,


Mary Fletcher
Electron Microscopist
Materials Eng. UBC
#309 - 6350 Stores Road
Vancouver, B.C. V6T 1Z4
Canada
Tel: 604-822-5648
Fax: 604-822-3619
email: maryflet-at-interchange.ubc.ca



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From: marie.cantino-at-uconn.edu
Date: Thu, 4 Nov 2010 11:09:09 -0500
Subject: [Microscopy] RE: EDS detection of Boron

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Thanks to everyone who took the time to respond to my (rather vague)
question about Boron detection. The question was elicited by a user
who would like to look at Boron distributions in plant tissue. The
estimated concentration was in the 100 ppm range. No information was
specified about the detector because we don't have it . . .yet.
However, from the feedback, it sounds like 100 ppm (which corresponds
to .01% by my calculation) is probably undetectable under any
condition. I'm not sure whether his estimate was for wet or dry
weight, but at best this might gain us (after drying) a factor of 8 or
so, still undetectable.

Marie

Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369


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From: ehaller-at-health.usf.edu
Date: Thu, 4 Nov 2010 13:02:19 -0500
Subject: [Microscopy] So long and Thanks

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Randy,

You will be missed by those of us who have taken advantage of your wisdom and experience over many years of participation on the Listserv. I do not think I am remiss in inviting you to re-join us in the future. You do not need to be at a scope to be a microscopist!
Thanks,
Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E - Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not represent the NIH. This message is not confidential and can be freely shared and reproduced.


________________________________
X-from: Randy Tindall {TindallR-at-missouri.edu}
Reply-To: Randy Tindall {TindallR-at-missouri.edu}

Hopefully, it will be a short break, Randy. I looked forward to your insightful posts. Enjoy your time off. You're a real resource to the whole of us! Please keep in touch!

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-cas.usf.edu
________________________________________
X-from: TindallR-at-missouri.edu [TindallR-at-missouri.edu]
Sent: Thursday, November 04, 2010 11:13 AM
To: Haller, Edward

Dear Collective,

I'm getting ready to take a break from the world of Electron Microscopy and just wanted to thank all the Denizens of the List who have helped rescue my hindquarters over the years. Seemed like no matter how deep the hole I was in, someone out there had the right rope to pull me out and the generosity to throw it to me.

This is a really great resource thanks to all of you, and, of course, our Friendly Neighborhood Sysop Nestor, who keeps it running smoothly. It has been a great pleasure to be part of it.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu
On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount=Week&NavType=Both&Type=TimePlan
Sons of Norway: http://www.sofn.com



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From: leunissen-at-aurion.nl
Date: Thu, 4 Nov 2010 13:11:54 -0500
Subject: [Microscopy] Re: So long and Thanks

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Randy,

I hope I can still add to the best wishes before Nestor will be forced to call it a day because of an email tsunami...
I am sure you will remember the knife breaker blues. Thanks a lot for great times.

All the best

Jan

On 5/11/2010, at 4:06 AM, TindallR-at-missouri.edu wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear Collective,
}
} I'm getting ready to take a break from the world of Electron Microscopy and just wanted to thank all the Denizens of the List who have helped rescue my hindquarters over the years. Seemed like no matter how deep the hole I was in, someone out there had the right rope to pull me out and the generosity to throw it to me.
}
} This is a really great resource thanks to all of you, and, of course, our Friendly Neighborhood Sysop Nestor, who keeps it running smoothly. It has been a great pleasure to be part of it.
}
} Cheers,
} Randy
}
} Randy Tindall
} Senior EM Specialist
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From: Jean.Underwood-at-Umassmed.edu
Date: Thu, 4 Nov 2010 19:38:13 -0500
Subject: [Microscopy] viaWWW: Electron microscope film use and source

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Email: Jean.Underwood-at-Umassmed.edu
Name: Jean Underwood

Organization: UMass Medical School

Title-Subject: [Filtered] Electron microscope film use and source

Message: Although the EM facility has primarily gone digital here, we
prefer to still use film for the detail it has for the type of images
we take. We will need to purchase our own film now as the facility
will no longer be supporting film, and I was wondering who currently
is using Kodak 4489 electron microscope film, 3 1/4 x 4, its general
availability, and what sources you are using to buy it from?
Thanks,
Jean Underwood
Cell Biology
UMass Medical School
Jean.Underwood-at-umassmed.edu
(508) 856-6021

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From: sllara-at-u.washington.edu
Date: Thu, 4 Nov 2010 19:40:58 -0500
Subject: [Microscopy] viaWWW: Best Wishes

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Name: Stephanie Lara

Organization: University of Washington

Title-Subject: [Filtered] Best Wishes

Message: We will miss the perspective and humor of "We Do Small".

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From: dsherman-at-purdue.edu
Date: Thu, 4 Nov 2010 21:08:07 -0500
Subject: [Microscopy] Re: viaWWW: Electron microscope film use and source

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Jean,

I am also a film person and will continue to be for any images that require
real resolution retention. Digital cannot equal it unless you have an 8K
camera. However, we have used Kodak SO-163 for many years. I think we got
on to it since it can be manipulated more by changing developer
concentration and time. Our structural biologists liked it for cryoTEM
images and we figured it would be helpful if both facilities used the same
film.

We get it from Aremac Holdings (formerly National Graphic Supply) in Albany,
NY 866-972-6464. I expect that they have the 4489 as well.

Debby


---
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy




On 11/4/10 8:40 PM, "Jean.Underwood-at-Umassmed.edu"
{Jean.Underwood-at-Umassmed.edu} wrote:

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} Title-Subject: [Filtered] Electron microscope film use and source
}
} Message: Although the EM facility has primarily gone digital here, we
} prefer to still use film for the detail it has for the type of images
} we take. We will need to purchase our own film now as the facility
} will no longer be supporting film, and I was wondering who currently
} is using Kodak 4489 electron microscope film, 3 1/4 x 4, its general
} availability, and what sources you are using to buy it from?
} Thanks,
} Jean Underwood
} Cell Biology
} UMass Medical School
} Jean.Underwood-at-umassmed.edu
} (508) 856-6021
}
} Login Host: 146.189.245.137
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From: oshel1pe-at-cmich.edu
Date: Fri, 5 Nov 2010 09:11:02 -0500
Subject: [Microscopy] Re: viaWWW: Electron microscope film use and source

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Jean,

We shoot Kodak 4489 in our CM-10, and Electron Microscopy Sciences,
SPI, Ted Pella, and Ladd Research also carry this film.

Phil

} This Question/Comment was submitted to the Microscopy Listserver
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} Email: Jean.Underwood-at-Umassmed.edu
} Name: Jean Underwood
}
} Organization: UMass Medical School
}
} Title-Subject: [Filtered] Electron microscope film use and source
}
} Message: Although the EM facility has primarily gone digital here, we
} prefer to still use film for the detail it has for the type of images
} we take. We will need to purchase our own film now as the facility
} will no longer be supporting film, and I was wondering who currently
} is using Kodak 4489 electron microscope film, 3 1/4 x 4, its general
} availability, and what sources you are using to buy it from?
} Thanks,
} Jean Underwood
} Cell Biology
} UMass Medical School
} Jean.Underwood-at-umassmed.edu
} (508) 856-6021

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: walter.bobrowski-at-pfizer.com
Date: Fri, 5 Nov 2010 13:11:35 -0500
Subject: [Microscopy] viaWWW: Bacterial Immunogold Protocol

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Title-Subject: [Filtered] Bacterial Immunogold Protocol

Message: Attempting pre-embedding immunogold of bacterial
suspensions. First round reveals complete absence of 12nm gold
particles (which worked for post-embedding IEM). Did we spin off the
particles (3000 rpm for 5 minutes) during pelleting? Does anyone have
an ideal protocol?

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6, 18 -- Subject: viaWWW: Bacterial Immunogold Protocol
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From: PhillipsT-at-missouri.edu
Date: Fri, 5 Nov 2010 13:16:14 -0500
Subject: [Microscopy] viaWWW: Bacterial Immunogold Protocol

Contents Retrieved from Microscopy Listserver Archives
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You wouldn't spin them off. I presume you are aiming for a surface antigen. Steric hinderance might block access to the antibody in pre-embedding. You could try a nanogold secondary and then use a gold enhancement step to maximize signal. Does a fluorescent-tagged secondary work at the LM level with this approach?




Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


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Email: walter.bobrowski-at-pfizer.com
Name: Walter Bobrowski

Organization: MSA

Title-Subject: [Filtered] Bacterial Immunogold Protocol

Message: Attempting pre-embedding immunogold of bacterial
suspensions. First round reveals complete absence of 12nm gold
particles (which worked for post-embedding IEM). Did we spin off the
particles (3000 rpm for 5 minutes) during pelleting? Does anyone have
an ideal protocol?

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From: Shea.Miller-at-AGR.GC.CA
Date: Fri, 5 Nov 2010 13:40:03 -0500
Subject: [Microscopy] LM: mounting media for FITC labelled samples

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Hello list!

I have a user that is using FITC-labelled antibodies for immunohistochemistry on plant tissues. We have been using "Aquaperm" mounting medium (Lipshaw/Immunon)... a bottle that is at least 10 years old, and works well. We would like to buy more, but it no longer seems to be available. Does anyone have a favourite hardening mounting medium that is good with FITC? I have tried ordering a couple of things that "seemed" to have the same specs, but they just don't work the same way.



Thanks in advance

shea


Dr. S. Shea Miller
ECORC | CRECO
Agriculture and Agri-Food Canada | Agriculture et Agroalimentaire Canada
960 Carling Avenue | 960, avenue Carling
Ottawa, ON K1A 0C6
E-mail Address / Adresse courriel shea.miller-at-agr.gc.ca
Telephone | Téléphone 613-759-1760
Facsimile | Télécopieur 613-759-1701
Teletypewriter | Téléimprimeur 613-773-2600
Government of Canada | Gouvernement du Canada



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From: William.F.Tivol-at-aero.org
Date: 11/02/2010 02:26 PM
Subject: [Microscopy] viaWWW: stabilizing biofilm?

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Dear Alice,
That's a really thick biofilm. Not only would it have not to be
disturbed, it probably would have to be kept wet. Perhaps the best way to
preserve it would be to use something like a petri dish as a cookie
cutter, push it through the biofilm, then slide a blade between the film
and its substrate (or cut through the substrate, depending on whether the
substrate is rock or sand, etc.). This should result in containing a
piece of the biofilm in relatively intact form, and a fixative could be
added at that point. It might also be possible to seal the petri dish and
transport the whole thing to your lab. Good luck.
Yours,
Bill



X-from: alice.dohnalkova-at-pnl.gov
To: William.F.Tivol-at-aero.org


Email: alice.dohnalkova-at-pnl.gov
Name: Alice Dohnalkova

Organization: Pacific Northwest National Laboratory

Title-Subject: [Filtered] stabilizing biofilm?

Message: Dear List,

Could you please recommend a method for stabilizing / immobilization
of a natural biofilm (typical thickness 5-10 mm) for EM processing? I
was told this biofilm doesn't hold together too well, and the
sampling site is pretty remote, so there is a good chance it would
get damaged by the time it will reach me.
The sampling team will have 2.5% glut available, but no means of
preparing / heating e.g. agarose or gelatin (that would be the idea
of putting a thin layer on top of a biofilm).

I'd really appreciate any suggestion.
Thanks!
Alice.

Alice Dohnalkova
Sr. Research Scientist
ENVIRONMENTAL MOLECULAR SCIENCES LABORATORY
Pacific Northwest National Laboratory
902 Battelle Boulevard
P.O. Box 999, MSIN K8-93
Richland, WA 99352 USA
Tel: 509-371-6515
Fax: 509-371-6242
Alice.dohnalkova-at-pnl.gov
www.pnl.gov


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From: William.F.Tivol-at-aero.org
Date: 11/02/2010 02:26 PM
Subject: [Microscopy] viaWWW: stabilizing biofilm?

Contents Retrieved from Microscopy Listserver Archives
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From: sjrobin-at-illinois.edu
Date: Fri, 5 Nov 2010 16:17:19 -0500
Subject: [Microscopy] TEM fixation of unicellular cyanobacteria

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Hello Out There--

I have made several attempts to fix and embed, for TEM, small pellets of
unicellular cyanobacteria. The pellets (in 'Eppy' tubes) are
E.M.-tissue-sized, no more than 0.5 mm on the narrow side. I fix using
2.0% form./2.5% glut. in 0.1 M cacodylate buffer for 4 hours in the
fridge, do a brief (10 minute) buffer rinse, fix 90 minutes in the dark
in 1% OsO4 in the same buffer, do another buffer rinse, then do an
ethanol dehydration, run through propylene oxide as a transitional
solvent, and embed in Epon 812. All standard for tissue and bacteria for
me for the past 28 years. But the fixation is horrendously bad, the
sections have holes in them that I could work around otherwise, and the
Epon appears dark (maybe too long in the osmium for this kind of
sample?). The first couple of times I tried this, the glut. came from
the vendor at the wrong concentration (I sent them the lot no. and they
told me I shouldn't have been sent that formulation). But with this last
prep I used what should have been good glut. (25% E.M. grade from
ampoules, diluted to 2.5%). And it looks just as bad. SEM samples (CHO
cells) that I used the same good glut. on looked fine.

If you have any tips or tricks I would very much appreciate your help.

Thank you

Scott

--
Microscopy Suite | Imaging Technology Group
Beckman Institute for Advanced Science and Technology
University of Illinois at Urbana-Champaign
405 North Mathews Avenue, Urbana IL 61801
217 265 5071 | 217 244 6219 (fax)
sjrobin-at-illinois.edu
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http://bugscope.beckman.illinois.edu




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From: gw265-at-cam.ac.uk
Date: Fri, 5 Nov 2010 17:21:42 -0500
Subject: [Microscopy] film casting: the non-Hollywood version

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Hi all,

Re: casting formvar or pioloform films for TEM grids - two questions:

(1) the old question of which slides are best for casting plastic films
has reared its head again. I've always found that a freshly-opened box of
precleaned slides is generally ok, but if the (presumably airtight)
plastic has been off the box more than about a day before, then the film
just won't come off the slide. Of course some brands are better than
others, but the newness seems to have an effect; particularly in warm,
humid places. It has been postulated that there is oil that picks up
humidity in the air, on the surface of these slides. Is this just a myth
I've been perpetrating all these years for the benefit of the slide
industry?

Does anyone know what the physical basis of the myth might be?


(2) How old is an old solution of 6% formvar? I recently cast some films
for slot grids that had the interesting property of forming hundreds of
small holes under the beam, in the first few minutes, then stabilising
(the holes didn't grow for a while), then ripping right across the film in
several places so that the ribbons of sections were the only things
holding it all together.

The films were pale yellow when they were cast earlier this week, carbon
coated with 2x 500ms pulses a day later, the beam was 120kV, 100um
condenser aperture, 2nd objective aperture in. Staining had been done the
day before looking, and the grids had been dried overnight with the lid of
the grid-box off. There was basically no dirt on the grids, and there were
3 ribbons per grid of cleanly-cut 50nm sections, covering most of the
slot.

The formvar solution was from January, had probably been used weekly since
then, and the hypothesis is that it goes off. I would suspect that the
concentration certainly changes with that much time with the lid off the
bottle. Any ideas?


thanks

Giselle Walker
University of Cambridge
UK

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From: leunissen-at-aurion.nl
Date: Fri, 5 Nov 2010 19:36:19 -0500
Subject: [Microscopy] viaWWW: Bacterial Immunogold Protocol

Contents Retrieved from Microscopy Listserver Archives
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Dear Walter and Tom,


yes, if this is about labelling a surface antigen it could very well be steric hinderance. In E coli LPS chains can be responsible. To reach antigens in the outer membrane we needed to use cryo ultramicrotomy, intact cell labelling did not work.

This was described in

Steric hindrance in immunolabelling. Voorhout WF, Leunissen-Bijvelt JJ, Leunissen JLM, Verkleij AJ.
J Microsc. 1986 Mar;141(Pt 3):303-10.


Good luck


Jan Leunissen
Aurion - Immunogold Reagents
http://www.aurion.nl


On 6/11/2010, at 7:16 AM, phillipst-at-missouri.edu wrote:

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} You wouldn't spin them off. I presume you are aiming for a surface antigen. Steric hinderance might block access to the antibody in pre-embedding. You could try a nanogold secondary and then use a gold enhancement step to maximize signal. Does a fluorescent-tagged secondary work at the LM level with this approach?
}
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} Thomas E. Phillips, Ph.D
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} Title-Subject: [Filtered] Bacterial Immunogold Protocol
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} Message: Attempting pre-embedding immunogold of bacterial
} suspensions. First round reveals complete absence of 12nm gold
} particles (which worked for post-embedding IEM). Did we spin off the
} particles (3000 rpm for 5 minutes) during pelleting? Does anyone have
} an ideal protocol?
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From: FMonson-at-wcupa.edu
Date: 11/02/2010 02:26 PM
Subject: [Microscopy] viaWWW: stabilizing biofilm?

Contents Retrieved from Microscopy Listserver Archives
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I agree with Bill on the sampling strategy, and only have one addition with respect to fixation.
If the biofilm is to be taken essentially intact, then I would suggest a double container (i.e., a peanut butter jar - large for Petri dish). In any case for the time up to shipping, store the Petri dish with biofilm in the closed container with the fixative set so that its fumes can penetrate the biofilm without touching it. We have found it helpful to fix microorganisms in the diffusing gas rather than in the added solution. After a few hours (overnight with OSO4 or HCHO) we can immerse in glut/buffer to finish and proceed. Especially with the OsO4, we find that the pm is stabilized(?) against frank destruction in subsequent steps.

Hoping I'm correct in your case,

Fred Monson
CMIRT
West Chester University
West Chester, PA, 19320
610-738-0437
http://cmirt.wcupa.edu
________________________________________
X-from: William.F.Tivol-at-aero.org [William.F.Tivol-at-aero.org]
Sent: Friday, November 05, 2010 4:20 PM
To: Monson, Frederick

Dear Alice,
That's a really thick biofilm. Not only would it have not to be
disturbed, it probably would have to be kept wet. Perhaps the best way to
preserve it would be to use something like a petri dish as a cookie
cutter, push it through the biofilm, then slide a blade between the film
and its substrate (or cut through the substrate, depending on whether the
substrate is rock or sand, etc.). This should result in containing a
piece of the biofilm in relatively intact form, and a fixative could be
added at that point. It might also be possible to seal the petri dish and
transport the whole thing to your lab. Good luck.
Yours,
Bill



X-from: alice.dohnalkova-at-pnl.gov
To: William.F.Tivol-at-aero.org


Email: alice.dohnalkova-at-pnl.gov
Name: Alice Dohnalkova

Organization: Pacific Northwest National Laboratory

Title-Subject: [Filtered] stabilizing biofilm?

Message: Dear List,

Could you please recommend a method for stabilizing / immobilization
of a natural biofilm (typical thickness 5-10 mm) for EM processing? I
was told this biofilm doesn't hold together too well, and the
sampling site is pretty remote, so there is a good chance it would
get damaged by the time it will reach me.
The sampling team will have 2.5% glut available, but no means of
preparing / heating e.g. agarose or gelatin (that would be the idea
of putting a thin layer on top of a biofilm).

I'd really appreciate any suggestion.
Thanks!
Alice.

Alice Dohnalkova
Sr. Research Scientist
ENVIRONMENTAL MOLECULAR SCIENCES LABORATORY
Pacific Northwest National Laboratory
902 Battelle Boulevard
P.O. Box 999, MSIN K8-93
Richland, WA 99352 USA
Tel: 509-371-6515
Fax: 509-371-6242
Alice.dohnalkova-at-pnl.gov
www.pnl.gov


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From: FMonson-at-wcupa.edu
Date: 11/02/2010 02:26 PM
Subject: [Microscopy] viaWWW: stabilizing biofilm?

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From: AJBowling-at-dow.com
Date: Sun, 7 Nov 2010 10:41:32 -0600
Subject: [Microscopy] TEM fixation of unicellular cyanobacteria

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Hi Scott,

There are a few things that I can think of that might help. For the
fixation, you might try fixing in suspension (not pelleted), with higher
concentration of glut (e.g. 6%), for longer (overnight at 4). Some
cyanobacteria make polysaccharide coatings that are difficult for
fixatives and dehydrants to penetrate. In a worst case, you might have
to resuspend and re-spin at each step to get decent fixation,
dehydration, and infiltration. As for the dark resin, maybe moving to a
fresh tube after osmication might help, since osmium can react with
plastics. Or maybe do the osmication in a glass scintillation vial or
test tube, then move back to the eppy for pelleting and dehydration,
etc.

Good luck,

Andy Bowling



_____________________
Andrew J Bowling, PhD
Dow AgroSciences
9330 Zionsville Rd
Indianapolis, IN 46268
317-337-3878



-----Original Message-----
X-from: sjrobin-at-illinois.edu [mailto:sjrobin-at-illinois.edu]
Sent: Friday, November 05, 2010 5:27 PM
To: Bowling, Andrew (AJ)

Hello Out There--

I have made several attempts to fix and embed, for TEM, small pellets of
unicellular cyanobacteria. The pellets (in 'Eppy' tubes) are
E.M.-tissue-sized, no more than 0.5 mm on the narrow side. I fix using
2.0% form./2.5% glut. in 0.1 M cacodylate buffer for 4 hours in the
fridge, do a brief (10 minute) buffer rinse, fix 90 minutes in the dark
in 1% OsO4 in the same buffer, do another buffer rinse, then do an
ethanol dehydration, run through propylene oxide as a transitional
solvent, and embed in Epon 812. All standard for tissue and bacteria for
me for the past 28 years. But the fixation is horrendously bad, the
sections have holes in them that I could work around otherwise, and the
Epon appears dark (maybe too long in the osmium for this kind of
sample?). The first couple of times I tried this, the glut. came from
the vendor at the wrong concentration (I sent them the lot no. and they
told me I shouldn't have been sent that formulation). But with this last
prep I used what should have been good glut. (25% E.M. grade from
ampoules, diluted to 2.5%). And it looks just as bad. SEM samples (CHO
cells) that I used the same good glut. on looked fine.

If you have any tips or tricks I would very much appreciate your help.

Thank you

Scott

--
Microscopy Suite | Imaging Technology Group
Beckman Institute for Advanced Science and Technology
University of Illinois at Urbana-Champaign
405 North Mathews Avenue, Urbana IL 61801
217 265 5071 | 217 244 6219 (fax)
sjrobin-at-illinois.edu
http://itg.beckman.illinois.edu
http://bugscope.beckman.illinois.edu




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24, 34 -- From AJBowling-at-dow.com Sun Nov 7 10:41:31 2010
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From: ZZhang-at-uwyo.edu
Date: Mon, 8 Nov 2010 09:34:23 -0600
Subject: [Microscopy] RE: TEM fixation of unicellular cyanobacteria

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Scott

For several years we had research students preparing isolated
cyanobacteria or lichens which contained cyanobacteria. The walls of
the vegetative cells could be a problem, often the heterocysts were
even worse and some storage bodies had a habit of dropping out (eg
polyphosphate).

Sometimes the only good results were obtained with KMnO4 fixation,
prolonged dehydration and embedding for extended periods in low
viscosity resins such as Spurr's. Our researchers got good results in
the end but it did seem like a bit of a black art.

Good luck

Malcolm

Malcolm Haswell
Imaging Suite
Faculty of Applied Sciences
University of Sunderland
SUNDERLAND
SR1 3SD
UK
email: malcolm.haswell-at-sunderland.ac.uk

----- Original Message -----
X-from: AJBowling-at-dow.com

Hi Scott:

We work with budding yeast for TEM analysis all the time. Yeast also has a thick cell wall, in a sense, similar to cyanobacteria.

The OsO4 cannot penetrate through the cell wall and you have to use enzyme to remove the cell wall before applying the OsO4. We use Zymolyase 100T for yeast cell wall.

As others suggested, potassium permanganate can be used as a alternative to OsO4. Here is a reference. Let me know if you need a PDF copy of the paper.

Yang H, Ren Q and Zhang Z (2006) Chromosome or chromatin condensation leads to meiosis or apoptosis in stationary yeast (Saccharomyces cerevisiae) cells. FEMS Yeast Res. 6(8):1254-1263.

Good luck,

Zhaojie


Zhaojie Zhang, Ph. D.
Director, Jenkins Microscopy Facility
University of Wyoming
Laramie, WY 82071
PHONE: 307-766-3038
FAX: 307-766-5625






} Hello Out There--
}
} I have made several attempts to fix and embed, for TEM, small
} pellets of
} unicellular cyanobacteria. The pellets (in 'Eppy' tubes) are
} E.M.-tissue-sized, no more than 0.5 mm on the narrow side. I fix
using
} 2.0% form./2.5% glut. in 0.1 M cacodylate buffer for 4 hours in the
} fridge, do a brief (10 minute) buffer rinse, fix 90 minutes in the
} darkin 1% OsO4 in the same buffer, do another buffer rinse, then
} do an
} ethanol dehydration, run through propylene oxide as a transitional
} solvent, and embed in Epon 812. All standard for tissue and
} bacteria for
} me for the past 28 years. But the fixation is horrendously bad, the
} sections have holes in them that I could work around otherwise,
} and the
} Epon appears dark (maybe too long in the osmium for this kind of
} sample?). The first couple of times I tried this, the glut. came from
} the vendor at the wrong concentration (I sent them the lot no. and
} theytold me I shouldn't have been sent that formulation). But with
} this last
} prep I used what should have been good glut. (25% E.M. grade from
} ampoules, diluted to 2.5%). And it looks just as bad. SEM samples
(CHO
} cells) that I used the same good glut. on looked fine.
}
} If you have any tips or tricks I would very much appreciate your
} help.
}
} Thank you
}
} Scott
}
} --
} Microscopy Suite | Imaging Technology Group
} Beckman Institute for Advanced Science and Technology
} University of Illinois at Urbana-Champaign
} 405 North Mathews Avenue, Urbana IL 61801
} 217 265 5071 | 217 244 6219 (fax)
} sjrobin-at-illinois.edu
} http://itg.beckman.illinois.edu
} http://bugscope.beckman.illinois.edu
}
}
}
}
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From: allan.mitchell-at-stonebow.otago.ac.nz
Date: Tue, 9 Nov 2010 00:59:16 -0600
Subject: [Microscopy] JEOL 840 SEM being disposed of

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi all

We are about to dismantle and dispose of the JEOL JSM 840 SEM here at
the University of Otago. If there are components that anybody would
like saved please let me know and we will try to rescue them. Freight
costs are the responsibility of the person making the request.

Have a great day

Allan




Allan Mitchell
Microscopy Otago - Electron Microscopy
c/- Department of Anatomy and Structural Biology
Otago School of Medical Sciences
P.O. Box 913
Dunedin
New Zealand

Phone (03) 479 5642 or 479 7301
Fax (03) 479 5086 or 479 7254

EM Centre: http://ocem.otago.ac.nz/
Confocal Centre: http://occm.otago.ac.nz/
NZ Microscopy Society: http://microscopynz.co.nz/




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From: bradbury.arcana-at-btinternet.com
Date: Tue, 9 Nov 2010 05:40:09 -0600
Subject: [Microscopy] viaWWW: SEM circuits- ISI TV Mini SEM

Contents Retrieved from Microscopy Listserver Archives
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Email: bradbury.arcana-at-btinternet.com
Name: david charles bradbury

Organization: retired geologist

Title-Subject: [Filtered] SEM circuits

Message: Morning Folks.
I am a retired mineralogist/geologist, recently I bought a S/H SEM,
sadly it was damaged by the previous owner, ie a few cables, I am
asking has anyone the circuits for the following SEM. ISI TV Mini SEM
? I am quite happy to pay for the circuits and postage to UK. Hope
someone can help.
Many Thanks...David Bradbury..UK.

Login Host: 81.132.11.226
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From: lesley.bechtold-at-jax.org
Date: Tue, 9 Nov 2010 14:49:39 -0600
Subject: [Microscopy] viaWWW: EM Facility Management

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Email: lesley.bechtold-at-jax.org
Name: Lesley Bechtold

Organization: The Jackson Laboratory

Title-Subject: [Filtered] EM Facility Management

Message: Last year, we carried out an online survey on EM Facility
Management. If anyone is intetested in the results, please email me
directly.

If anyone would like to parcticipate in this year's survey, I'd be
happy to send you the link.

Lesley Bechtold

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The Jackson Laboratory
Bar Harbor ME 04609

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From: Lindsay.P.Keller-at-nasa.gov
Date: Tue, 9 Nov 2010 15:42:30 -0600
Subject: [Microscopy] viaWWW: mapping montage misfit with Noran System Six

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Email: Lindsay.P.Keller-at-nasa.gov
Name: L. Keller

Organization: NASA

Title-Subject: [Filtered] mapping montage misfit with Noran System Six

Message: We are using a Noran SDD on our FEG SEM to produce spectrum
images of rock thin sections. One issue we are having is that in
assembling montages of large areas, there are gaps between the
individual component images - we've double checked the calibration a
couple times, but still have an issue. Has anyone else encountered
this problem? Thanks.

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From: donovan-at-uoregon.edu
Date: Tue, 9 Nov 2010 16:03:53 -0600
Subject: [Microscopy] Re: viaWWW: mapping montage misfit with Noran

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Hi Lindsay,
I've been doing some NSS montage mapping lately also and find that
the mismatch is very sensitive to the scan rotation. In fact on my
SX100 I have documented small but steady changes in scan rotation as
a function of mag. The eye is very sensitive to this error.

You therefore must calibrate at the same mag that you will be
acquiring the images for best results. Are you doing that?
john

At 01:45 PM 11/9/2010, you wrote:



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From: rcommon-at-msu.edu
Date: Tue, 9 Nov 2010 17:51:26 -0600
Subject: [Microscopy] GMA resin

Contents Retrieved from Microscopy Listserver Archives
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GMA kits for resins such as Immuno-Bed, JB-4 and Technivit have become
very expensive. I would like to try mixing my own resin from
components. Does anybody out there have a tried and true recipe for a
resin with properties similar to these commercial products? Since I
often work with large pigmented specimens, I need something that can
polymerize at low temperature without UV light. I need something with
good cutting properties but Immuno staining is not important. I would
be very grateful for any help.

Ralph Common

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From: kamlennon-at-yahoo.com
Date: Tue, 9 Nov 2010 20:44:04 -0600
Subject: [Microscopy] Osmium safety advice etc.

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi Listers,

I'm gearing up to teach introductory electron microscopy to undergraduate
biology majors again this Spring and have hit a few safety road blocks on which
I'd greatly appreciate your advice. Yes, undergrads in an EM lab is a big, red
safety flag on it's own!

First, I'm on my own with this, as there is no one else for miles who has any
expertise with EM and we don't have a dedicated health and safety unit at my
University. I am very spoiled in that I've always been an EM user with dedicated
staff and safety personnel within reach. So, the first thing that I need is a
full face respirator in the event that someone drops Osmium because it's all up
to me to clean it up. My first question is, do I need filtered air or supplied
air? Any other suggestions?

Secondly, I have a faint memory of there being a way to reduce the toxicity of
Osmium waste by binding the osmium to oil. Am I hallucinating or does someone
have some tips on doing this? We're trying to reduce the toxicity and save the
University some cash in waste disposal fees because it will be cheaper to deal
with less toxic waste.


Now, a couple of other question related to the class. I know that I'm going to
run into some trouble when I tell our budget keepers that I've got to order new
supplies because 2 year old glut is not acceptable. EM is expensive! We have a
heavy focus on field science, so the expense of this class comes as a shock.
Does anyone have any insight as to the "shelf life" of unopened ampules of
glutaraldehyde? Maybe this would be a better buy this year.

Lastly, as a plant person, I'm struggling with how to get my students experience
with animal tissue because I lost my colleague who used to sacrifice rats for
his research and lend me some tissue for my class. I do have a connection to a
local vet and am wondering if anyone has any advice as to whether having him pop
some of the tissues that he harvests from his patients into vials of Karnovsky's
would be a good option. My students may get a lot of cat testicles and ovaries,
but at least they'd get some animal tissue!

Thanks in advance for any advice you can lend. I'm sure that I'll be in touch as
I muddle my way through another semester of enlightening undergrads.
-Kristen Lennon

Kristen Lennon, Ph.D.
Department of Biology
Frostburg State University
Frostburg, MD 21532
kalennon-at-frostburg.edu





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From: dsherman-at-purdue.edu
Date: Tue, 9 Nov 2010 22:18:09 -0600
Subject: [Microscopy] Re: Osmium safety advice etc.

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Kristen,

First thing is don't panic. Not all undergrads are disaster in the making.
Lots of grad students are the same....I have engineering students who have
never used a pipette or pH meter let alone worked in or near a fume hood!
I've also had undergrads who have been fantastic students in all respects.

A couple of mandatory rules will help, including handling all chemicals in
hoods AT ALL TIMES! This should limit the inevitable small spills to at
least fairly safe confines. I also make students work on enamel trays in the
hood to further confine spills. And of course all students need to wear
gloves when working with chemicals of any kind.

Regarding osmium...fill a 1 liter polypropylene bottle (preferably
wide-mouth) about 1/3 full with vegetable oil...the stuff you use for
cooking. Add osmium waste. The osmium will react with the oil which
renders it much less reactive. We always treat osmium waste this way to aid
safer disposal.

Don't worry about the glutaraldehyde. As long as it has been sealed in vials
it should be fine. Remember this is a basic class and you are not trying to
optimize sample preparation. Only problem I have had with older
glutaraldehyde is when the concentration is above 25%. In this case the
glut was cloudy when diluted. However, we do try to use up glutaraldehyde
within a year just to avoid potential problems. We also try to use it
within a week once the vial is opened and the glutaraldehyde is diluted.

As to animal tissue, is anyone at your university working with mice or zebra
fish who could provide some tissue? Perhaps someone is working with
fertilized eggs and you can get a chicken embryo.

Good luck...it is always a challenge teaching lab courses but can be very
satisfying as well if you actually turn on some of the students to science
research...especially microscopy.

Debby


---
Debby Sherman, Director Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www.ag.purdue.edu/facilities/microscopy









On 11/9/10 9:46 PM, "kamlennon-at-yahoo.com" {kamlennon-at-yahoo.com} wrote:

}
}
}
} ----------------------------------------------------------------------------
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} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi Listers,
}
} I'm gearing up to teach introductory electron microscopy to undergraduate
} biology majors again this Spring and have hit a few safety road blocks on
} which
} I'd greatly appreciate your advice. Yes, undergrads in an EM lab is a big, red
} safety flag on it's own!
}
} First, I'm on my own with this, as there is no one else for miles who has any
} expertise with EM and we don't have a dedicated health and safety unit at my
} University. I am very spoiled in that I've always been an EM user with
} dedicated
} staff and safety personnel within reach. So, the first thing that I need is a
} full face respirator in the event that someone drops Osmium because it's all
} up
} to me to clean it up. My first question is, do I need filtered air or supplied
} air? Any other suggestions?
}
} Secondly, I have a faint memory of there being a way to reduce the toxicity of
} Osmium waste by binding the osmium to oil. Am I hallucinating or does someone
} have some tips on doing this? We're trying to reduce the toxicity and save the
} University some cash in waste disposal fees because it will be cheaper to deal
} with less toxic waste.
}
}
} Now, a couple of other question related to the class. I know that I'm going to
} run into some trouble when I tell our budget keepers that I've got to order
} new
} supplies because 2 year old glut is not acceptable. EM is expensive! We have a
} heavy focus on field science, so the expense of this class comes as a shock.
} Does anyone have any insight as to the "shelf life" of unopened ampules of
} glutaraldehyde? Maybe this would be a better buy this year.
}
} Lastly, as a plant person, I'm struggling with how to get my students
} experience
} with animal tissue because I lost my colleague who used to sacrifice rats for
} his research and lend me some tissue for my class. I do have a connection to a
} local vet and am wondering if anyone has any advice as to whether having him
} pop
} some of the tissues that he harvests from his patients into vials of
} Karnovsky's
} would be a good option. My students may get a lot of cat testicles and
} ovaries,
} but at least they'd get some animal tissue!
}
} Thanks in advance for any advice you can lend. I'm sure that I'll be in touch
} as
} I muddle my way through another semester of enlightening undergrads.
} -Kristen Lennon
}
} Kristen Lennon, Ph.D.
} Department of Biology
} Frostburg State University
} Frostburg, MD 21532
} kalennon-at-frostburg.edu
}
}
}
}
}
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} 12, 28 -- Date: Tue, 9 Nov 2010 18:44:02 -0800 (PST)
} 12, 28 -- From: Kristen Lennon {kamlennon-at-yahoo.com}
} 12, 28 -- Subject: Osmium safety advice etc.
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From: nizets2-at-yahoo.com
Date: Wed, 10 Nov 2010 03:29:41 -0600
Subject: [Microscopy] Osmium safety advice etc.

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi Kristen!

The toxicity of a product is a fonction of its concentration and the amount that
is absorbed, so use diluted solutions and don't let the students play with large
solutions and you have made a big step in the security. The osmium solution must
never leave the hood, so I don't see why you would need a full face protection.
If a few drops have left the hood, safety (closed) goggles should be enough.
Otherwise Debby gave good advices. To neutralize osmium one needs unsaturated
fatty acids, generally sunflower oil is cheap enough to be used.
As for the samples your idea is good but the vet should better slice the organs
or alternatively the organs may be conserved in ice (for 1-2 hours should not be
too bad). Then you can slice them and fix them yourself (or the student can do
it). Skin should also no be too hard to get from a vet and the histology is
interesting and not too complicated.

Stephane


 


----- Original Message ----
X-from: "kamlennon-at-yahoo.com" {kamlennon-at-yahoo.com}
To: nizets2-at-yahoo.com
Sent: Wed, November 10, 2010 3:48:20 AM

Hi Listers,

I'm gearing up to teach introductory electron microscopy to undergraduate
biology majors again this Spring and have hit a few safety road blocks on which
I'd greatly appreciate your advice. Yes, undergrads in an EM lab is a big, red
safety flag on it's own!

First, I'm on my own with this, as there is no one else for miles who has any
expertise with EM and we don't have a dedicated health and safety unit at my
University. I am very spoiled in that I've always been an EM user with dedicated

staff and safety personnel within reach. So, the first thing that I need is a
full face respirator in the event that someone drops Osmium because it's all up
to me to clean it up. My first question is, do I need filtered air or supplied
air? Any other suggestions?

Secondly, I have a faint memory of there being a way to reduce the toxicity of
Osmium waste by binding the osmium to oil. Am I hallucinating or does someone
have some tips on doing this? We're trying to reduce the toxicity and save the
University some cash in waste disposal fees because it will be cheaper to deal
with less toxic waste.


Now, a couple of other question related to the class. I know that I'm going to
run into some trouble when I tell our budget keepers that I've got to order new
supplies because 2 year old glut is not acceptable. EM is expensive! We have a
heavy focus on field science, so the expense of this class comes as a shock.
Does anyone have any insight as to the "shelf life" of unopened ampules of
glutaraldehyde? Maybe this would be a better buy this year.

Lastly, as a plant person, I'm struggling with how to get my students experience

with animal tissue because I lost my colleague who used to sacrifice rats for
his research and lend me some tissue for my class. I do have a connection to a
local vet and am wondering if anyone has any advice as to whether having him pop

some of the tissues that he harvests from his patients into vials of Karnovsky's

would be a good option. My students may get a lot of cat testicles and ovaries,
but at least they'd get some animal tissue!

Thanks in advance for any advice you can lend. I'm sure that I'll be in touch as

I muddle my way through another semester of enlightening undergrads.
-Kristen Lennon

Kristen Lennon, Ph.D.
Department of Biology
Frostburg State University
Frostburg, MD 21532
kalennon-at-frostburg.edu


     


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From: oshel1pe-at-cmich.edu
Date: Wed, 10 Nov 2010 08:33:29 -0600
Subject: [Microscopy] Re: Osmium safety advice etc.

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Kristen,

First, to echo other responders, don't worry
about teaching to undergrads. We have a
microscopy major within our Biology B.Sc. program
and routinely teach TEM and SEM to undergrads.
I've never found that undergrads are a big safety
problem. (Engineering grad students, now ...)
Teach proper techniques (not over-done panicky
Safety Committee Ohmygodosmium!) and they'll be
fine. Do emphasize that OsO4 is highly volatile
and *must* be used in a hood. Show them a
blackened OsO4 storage container and the inside
of your fixative 'frig. They'll get the point.
Keep a spill kit handy and show them how to use it.
OsO4 waste: as others have written, use vegetable
oil in a plastic container (not glass - glass
breaks, then you have a nasty mess), but find the
cheap highly polyunsaturated oil, like Canola.
The more unsaturated the oil, the more it binds
up OsO4. Also, add kitty litter to the waste
container. This keeps the oil from sloshing
around and increases the surface area of oil
available to bind OsO4.
And! Have a separate wide-mouth container for
solid OsO4 contaminated waste - transfer
pipettes, gloves, etc.
Emergency respirator - filtered is fine IF it's
the right filter. But mostly, have kitty litter
and oil handy to dump on any spill. But, if the
OsO4 is kept in the hood, then any spill should
be in the hood - preferably in a tray - so a
respirator really isn't necessary.

Re: samples. You can get away with using tissues
from a local vet or other people's research.
Unless you have an active animal-care committee
and animal research protocols. Then you'll likely
discover that you must have your own animal
tissue protocols even if you are using samples
taken from animals used in research or brought in
from off campus.
But.
If you use insects like crickets - used for
reptile food - or the cockroaches and spiders
hanging around your building, then you have no
issues with animal care committees, animal use
protocols (there are laws and regulations
involved here beyond your campus regs) and so on.
(Or isopods - pillbugs and sowbugs - or worms or
etc.) Fruitflies.
Plus, you get really neat specimens:
contracted/relaxed muscles, Malpighian tubules,
venom and silk glands, and lots more.
And there are lots of EM images in the literature
and the 15 volume "Microscopic Anatomy of
Invertebrates" published by Wiley-Liss and Miriam
Rothschild's beautiful "Insect Tissues via the
Flea".
Or, brine shrimp (Artemia) are easy to culture.
All the specimens for this would be free.

Note: just dumping testicles and ovaries into a
jar of Karnovsky's would be a good exercise in
poor preservation and why one minces tissue for
EM fixation.

Phil

} Hi Listers,
}
} I'm gearing up to teach introductory electron microscopy to undergraduate
} biology majors again this Spring and have hit a
} few safety road blocks on which
} I'd greatly appreciate your advice. Yes, undergrads in an EM lab is a big, red
} safety flag on it's own!
}
} First, I'm on my own with this, as there is no one else for miles who has any
} expertise with EM and we don't have a dedicated health and safety unit at my
} University. I am very spoiled in that I've
} always been an EM user with dedicated
} staff and safety personnel within reach. So, the first thing thatÝI need is a
} full face respirator in the event that someone
} drops Osmium because it's all up
} to me to clean it up. My first question is, do I need filtered air or supplied
} air? Any other suggestions?
}
} Secondly, I have a faint memory of there being a way to reduce the toxicity of
} Osmium waste by binding the osmium to oil. Am I hallucinating or does someone
} have some tips on doing this? We're trying to reduce the toxicity and save the
} University some cash in waste disposal fees because it will be cheaper to deal
} with less toxic waste.
}
}
} Now, a couple of other question related to the class. I know that I'm going to
} run into some trouble when I tell our budget
} keepers that I've got to order new
} supplies because 2 year old glut is not acceptable. EM is expensive! We have a
} heavy focus on field science, so the expense of this class comes as a shock.
} Does anyone have any insight as to the "shelf life" of unopened ampules of
} glutaraldehyde? Maybe this would be a better buy this year.
}
} Lastly, as a plant person, I'm struggling with
} how toÝget my students experience
} with animal tissueÝbecause I lost my colleague whoÝused to sacrificeÝrats for
} his research and lend me some tissue for my class. I do have a connection to a
} local vet and am wondering if anyone has any
} advice as to whether having him pop
} some of the tissues that he harvests from his
} patients into vials of Karnovsky's
} would be a good option. My students may get a
} lot of cat testicles and ovaries,
} but at least they'd get some animal tissue!
}
} Thanks in advance for any advice you can lend.
} I'm sure that I'll be in touch as
} I muddle my way through another semester of enlightening undergrads.
} -Kristen Lennon
}
} Kristen Lennon, Ph.D.
} Department of Biology
} Frostburg State University
} Frostburg, MD 21532
} kalennon-at-frostburg.edu

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576


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From: David.Patton-at-uwe.ac.uk
Date: Wed, 10 Nov 2010 09:42:44 -0600
Subject: [Microscopy] Re: Osmium safety advice etc.

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I would like to broaden the topic a little and request advice.

Osmium. I have been hording unreacted waste in Kilner/Mason jars in my fridge. I suppose that when it is reacted with oil then it is no longer volatile. Is this mixture in a polypropylene bottle with an ordinary screw lid a satisfactory container to pass on to a waste disposal operative?

Supplementary question: Phil - Kitty litter - I presume you mean the mineral type - how much do you use? Eg 10% of the bottle's volume?

Waste gluraraldehyde in buffer. This will probably still be volatile. What is a satisfactory container to pass on to a waste disposal operative?

Dave

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: 10 November 2010 14:38
To: David Patton

Kristen,

First, to echo other responders, don't worry
about teaching to undergrads. We have a
microscopy major within our Biology B.Sc. program
and routinely teach TEM and SEM to undergrads.
I've never found that undergrads are a big safety
problem. (Engineering grad students, now ...)
Teach proper techniques (not over-done panicky
Safety Committee Ohmygodosmium!) and they'll be
fine. Do emphasize that OsO4 is highly volatile
and *must* be used in a hood. Show them a
blackened OsO4 storage container and the inside
of your fixative 'frig. They'll get the point.
Keep a spill kit handy and show them how to use it.
OsO4 waste: as others have written, use vegetable
oil in a plastic container (not glass - glass
breaks, then you have a nasty mess), but find the
cheap highly polyunsaturated oil, like Canola.
The more unsaturated the oil, the more it binds
up OsO4. Also, add kitty litter to the waste
container. This keeps the oil from sloshing
around and increases the surface area of oil
available to bind OsO4.
And! Have a separate wide-mouth container for
solid OsO4 contaminated waste - transfer
pipettes, gloves, etc.
Emergency respirator - filtered is fine IF it's
the right filter. But mostly, have kitty litter
and oil handy to dump on any spill. But, if the
OsO4 is kept in the hood, then any spill should
be in the hood - preferably in a tray - so a
respirator really isn't necessary.

Re: samples. You can get away with using tissues
from a local vet or other people's research.
Unless you have an active animal-care committee
and animal research protocols. Then you'll likely
discover that you must have your own animal
tissue protocols even if you are using samples
taken from animals used in research or brought in
from off campus.
But.
If you use insects like crickets - used for
reptile food - or the cockroaches and spiders
hanging around your building, then you have no
issues with animal care committees, animal use
protocols (there are laws and regulations
involved here beyond your campus regs) and so on.
(Or isopods - pillbugs and sowbugs - or worms or
etc.) Fruitflies.
Plus, you get really neat specimens:
contracted/relaxed muscles, Malpighian tubules,
venom and silk glands, and lots more.
And there are lots of EM images in the literature
and the 15 volume "Microscopic Anatomy of
Invertebrates" published by Wiley-Liss and Miriam
Rothschild's beautiful "Insect Tissues via the
Flea".
Or, brine shrimp (Artemia) are easy to culture.
All the specimens for this would be free.

Note: just dumping testicles and ovaries into a
jar of Karnovsky's would be a good exercise in
poor preservation and why one minces tissue for
EM fixation.

Phil

} Hi Listers,
}
} I'm gearing up to teach introductory electron microscopy to undergraduate
} biology majors again this Spring and have hit a
} few safety road blocks on which
} I'd greatly appreciate your advice. Yes, undergrads in an EM lab is a big, red
} safety flag on it's own!
}
} First, I'm on my own with this, as there is no one else for miles who has any
} expertise with EM and we don't have a dedicated health and safety unit at my
} University. I am very spoiled in that I've
} always been an EM user with dedicated
} staff and safety personnel within reach. So, the first thing thatÝI need is a
} full face respirator in the event that someone
} drops Osmium because it's all up
} to me to clean it up. My first question is, do I need filtered air or supplied
} air? Any other suggestions?
}
} Secondly, I have a faint memory of there being a way to reduce the toxicity of
} Osmium waste by binding the osmium to oil. Am I hallucinating or does someone
} have some tips on doing this? We're trying to reduce the toxicity and save the
} University some cash in waste disposal fees because it will be cheaper to deal
} with less toxic waste.
}
}
} Now, a couple of other question related to the class. I know that I'm going to
} run into some trouble when I tell our budget
} keepers that I've got to order new
} supplies because 2 year old glut is not acceptable. EM is expensive! We have a
} heavy focus on field science, so the expense of this class comes as a shock.
} Does anyone have any insight as to the "shelf life" of unopened ampules of
} glutaraldehyde? Maybe this would be a better buy this year.
}
} Lastly, as a plant person, I'm struggling with
} how toÝget my students experience
} with animal tissueÝbecause I lost my colleague whoÝused to sacrificeÝrats for
} his research and lend me some tissue for my class. I do have a connection to a
} local vet and am wondering if anyone has any
} advice as to whether having him pop
} some of the tissues that he harvests from his
} patients into vials of Karnovsky's
} would be a good option. My students may get a
} lot of cat testicles and ovaries,
} but at least they'd get some animal tissue!
}
} Thanks in advance for any advice you can lend.
} I'm sure that I'll be in touch as
} I muddle my way through another semester of enlightening undergrads.
} -Kristen Lennon
}
} Kristen Lennon, Ph.D.
} Department of Biology
} Frostburg State University
} Frostburg, MD 21532
} kalennon-at-frostburg.edu

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576


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From: oshel1pe-at-cmich.edu
Date: Wed, 10 Nov 2010 10:08:49 -0600
Subject: [Microscopy] Osmium safety advice etc. CORRECT STORAGE

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dave,

} I would like to broaden the topic a little and request advice.
}
} Osmium. I have been hording unreacted waste in
} Kilner/Mason jars in my fridge. I suppose that
} when it is reacted with oil then it is no longer
} volatile. Is this mixture in a polypropylene
} bottle with an ordinary screw lid a satisfactory
} container to pass on to a waste disposal
} operative?

No. The osmium is volatile enough to escape a
(tightly) capped jar. The inside of your 'fridge
is pretty black from osmium, I imagine. Parafilm
(or what you have in the UK) the ever-loving the
cap of the jars.

} Supplementary question: Phil - Kitty litter - I
} presume you mean the mineral type - how much do
} you use? Eg 10% of the bottle's volume?

Yes, the clay type. This is less important than
is using something absorbent that the oil will
cling to. This is what increases the effective
surface area of the oil, making more of it
available to the osmium.
I eyeball a layer a couple of centimeters deep in
a US gallon/4L jug, then add around 200 - 500 mL
or so of oil.
As waste osmium accumulates, I'll add oil and maybe litter if it looks needed.

} Waste gluraraldehyde in buffer. This will
} probably still be volatile. What is a
} satisfactory container to pass on to a waste
} disposal operative?

A tightly capped plastic bottle. Glut is less
volatile than osmium, but Parafilming the cap
isn't a bad idea.

Mind, though, this can also depend on your local
laws and regulations, and the local hazardous
waste disposal people. They may have other ideas,
in which case you must do things their way.
Note that accumulating hazardous waste can be a
particular problem. Here in the US, there are
laws about how much waste of what kind can be
accumulated in a local area - like an individual
lab - and in a waste collection site and so on up
the ladder to the waste disposal/processing area.
I'd think the UK has similar laws. And in what
the waste is accumulated.
Your 'fridge hoard of unreacted waste may be a serious problem in this regard.

Phil

} Dave
}
} -----Original Message-----
} From: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
} Sent: 10 November 2010 14:38
} To: David Patton
} Subject: [Microscopy] Re: Osmium safety advice etc.
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576


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12, 33 -- Subject: RE: [Microscopy] Re: Osmium safety advice etc. CORRECT STORAGE
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From: PhillipsT-at-missouri.edu
Date: Wed, 10 Nov 2010 10:10:10 -0600
Subject: [Microscopy] Re: Osmium safety advice etc.

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

First, let me strongly suggest you not store unreacted waste in Mason jars in your refrigerator. My experience is no single jar seal is sufficient to contain osmium vapor. I store my 2% osmium and osmium waste in my fume hood. My 2% osmium stock is in a tightly sealed orange-cap Schott bottle. The orange cap has gone completely black. The entire Schott bottle is kept inside a clear plastic polycarbonate jar. The polycarbonate has gone completely black. I have seen, in other labs in the past, refrigerators with blackened walls due to storage of osmium as you suggest you do. As a general rule, store your osmium in double containers and keep them in a fume hood.

Secondly, it is not allowed at my University for an investigator to "process" hazardous waste without a written protocol that has been approved. I can't simply put my osmium waste in oil such as Debbie suggests. I think I once asked about that and was told not to do that. I don't know the rationale behind this denial.




Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: David.Patton-at-uwe.ac.uk [mailto:David.Patton-at-uwe.ac.uk]
Sent: Wednesday, November 10, 2010 9:44 AM
To: Phillips, Thomas E.

I would like to broaden the topic a little and request advice.

Osmium. I have been hording unreacted waste in Kilner/Mason jars in my fridge. I suppose that when it is reacted with oil then it is no longer volatile. Is this mixture in a polypropylene bottle with an ordinary screw lid a satisfactory container to pass on to a waste disposal operative?

Supplementary question: Phil - Kitty litter - I presume you mean the mineral type - how much do you use? Eg 10% of the bottle's volume?

Waste gluraraldehyde in buffer. This will probably still be volatile. What is a satisfactory container to pass on to a waste disposal operative?

Dave

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: 10 November 2010 14:38
To: David Patton

Kristen,

First, to echo other responders, don't worry
about teaching to undergrads. We have a
microscopy major within our Biology B.Sc. program
and routinely teach TEM and SEM to undergrads.
I've never found that undergrads are a big safety
problem. (Engineering grad students, now ...)
Teach proper techniques (not over-done panicky
Safety Committee Ohmygodosmium!) and they'll be
fine. Do emphasize that OsO4 is highly volatile
and *must* be used in a hood. Show them a
blackened OsO4 storage container and the inside
of your fixative 'frig. They'll get the point.
Keep a spill kit handy and show them how to use it.
OsO4 waste: as others have written, use vegetable
oil in a plastic container (not glass - glass
breaks, then you have a nasty mess), but find the
cheap highly polyunsaturated oil, like Canola.
The more unsaturated the oil, the more it binds
up OsO4. Also, add kitty litter to the waste
container. This keeps the oil from sloshing
around and increases the surface area of oil
available to bind OsO4.
And! Have a separate wide-mouth container for
solid OsO4 contaminated waste - transfer
pipettes, gloves, etc.
Emergency respirator - filtered is fine IF it's
the right filter. But mostly, have kitty litter
and oil handy to dump on any spill. But, if the
OsO4 is kept in the hood, then any spill should
be in the hood - preferably in a tray - so a
respirator really isn't necessary.

Re: samples. You can get away with using tissues
from a local vet or other people's research.
Unless you have an active animal-care committee
and animal research protocols. Then you'll likely
discover that you must have your own animal
tissue protocols even if you are using samples
taken from animals used in research or brought in
from off campus.
But.
If you use insects like crickets - used for
reptile food - or the cockroaches and spiders
hanging around your building, then you have no
issues with animal care committees, animal use
protocols (there are laws and regulations
involved here beyond your campus regs) and so on.
(Or isopods - pillbugs and sowbugs - or worms or
etc.) Fruitflies.
Plus, you get really neat specimens:
contracted/relaxed muscles, Malpighian tubules,
venom and silk glands, and lots more.
And there are lots of EM images in the literature
and the 15 volume "Microscopic Anatomy of
Invertebrates" published by Wiley-Liss and Miriam
Rothschild's beautiful "Insect Tissues via the
Flea".
Or, brine shrimp (Artemia) are easy to culture.
All the specimens for this would be free.

Note: just dumping testicles and ovaries into a
jar of Karnovsky's would be a good exercise in
poor preservation and why one minces tissue for
EM fixation.

Phil

} Hi Listers,
}
} I'm gearing up to teach introductory electron microscopy to undergraduate
} biology majors again this Spring and have hit a
} few safety road blocks on which
} I'd greatly appreciate your advice. Yes, undergrads in an EM lab is a big, red
} safety flag on it's own!
}
} First, I'm on my own with this, as there is no one else for miles who has any
} expertise with EM and we don't have a dedicated health and safety unit at my
} University. I am very spoiled in that I've
} always been an EM user with dedicated
} staff and safety personnel within reach. So, the first thing thatÝI need is a
} full face respirator in the event that someone
} drops Osmium because it's all up
} to me to clean it up. My first question is, do I need filtered air or supplied
} air? Any other suggestions?
}
} Secondly, I have a faint memory of there being a way to reduce the toxicity of
} Osmium waste by binding the osmium to oil. Am I hallucinating or does someone
} have some tips on doing this? We're trying to reduce the toxicity and save the
} University some cash in waste disposal fees because it will be cheaper to deal
} with less toxic waste.
}
}
} Now, a couple of other question related to the class. I know that I'm going to
} run into some trouble when I tell our budget
} keepers that I've got to order new
} supplies because 2 year old glut is not acceptable. EM is expensive! We have a
} heavy focus on field science, so the expense of this class comes as a shock.
} Does anyone have any insight as to the "shelf life" of unopened ampules of
} glutaraldehyde? Maybe this would be a better buy this year.
}
} Lastly, as a plant person, I'm struggling with
} how toÝget my students experience
} with animal tissueÝbecause I lost my colleague whoÝused to sacrificeÝrats for
} his research and lend me some tissue for my class. I do have a connection to a
} local vet and am wondering if anyone has any
} advice as to whether having him pop
} some of the tissues that he harvests from his
} patients into vials of Karnovsky's
} would be a good option. My students may get a
} lot of cat testicles and ovaries,
} but at least they'd get some animal tissue!
}
} Thanks in advance for any advice you can lend.
} I'm sure that I'll be in touch as
} I muddle my way through another semester of enlightening undergrads.
} -Kristen Lennon
}
} Kristen Lennon, Ph.D.
} Department of Biology
} Frostburg State University
} Frostburg, MD 21532
} kalennon-at-frostburg.edu

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576


==============================Original Headers==============================
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19, 36 -- From: David Patton {David.Patton-at-uwe.ac.uk}
19, 36 -- Subject: RE: [Microscopy] Re: Osmium safety advice etc. CORRECT STORAGE
19, 36 -- CONTAINERS FOR WASTE OSMIUM AND GLUTARALDEHYDE
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33, 33 -- From PhillipsT-at-missouri.edu Wed Nov 10 10:10:10 2010
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From: ben.micklem-at-pharm.ox.ac.uk
Date: Wed, 10 Nov 2010 10:18:23 -0600
Subject: [Microscopy] Re: Osmium safety advice etc. CORRECT STORAGE

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Before we take our osmium waste to our Safety Office we store it with
sawdust and oil in a plastic bottle (for the same purpose as cat litter).
About 50-75% of the 1 litre bottle is filled with sawdust, and enough
vegetable oil is poured in to generously cover the sawdust (which compacts
down a bit)- no real reason why we use these quantities, but they seem
sensible amounts.

We also have a dedicated bin for osmium-contaminated gloves, disposable
pipettes and benchcoat; after they have been left for at least 12 hours in
the hood for the osmium to evaporate. I think this bin is emptied into the
normal lab waste bins, but it is contained within black bags and treated
more sensitively to prevent exposure of the staff emptying bins.


Ben


--
Imaging Technician
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271866
{http://mrcanu.pharm.ox.ac.uk/}




On 10/11/2010 15:51, "David.Patton-at-uwe.ac.uk" {David.Patton-at-uwe.ac.uk}
wrote:


} I would like to broaden the topic a little and request advice.
}
} Osmium. I have been hording unreacted waste in Kilner/Mason jars in my
} fridge. I suppose that when it is reacted with oil then it is no longer
} volatile. Is this mixture in a polypropylene bottle with an ordinary
} screw lid a satisfactory container to pass on to a waste disposal
} operative?
}
} Supplementary question: Phil - Kitty litter - I presume you mean the
} mineral type - how much do you use? Eg 10% of the bottle's volume?
}
} Waste gluraraldehyde in buffer. This will probably still be volatile.
} What is a satisfactory container to pass on to a waste disposal operative?
}
} Dave



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From: oshel1pe-at-cmich.edu
Date: Wed, 10 Nov 2010 10:24:13 -0600
Subject: [Microscopy] RE: Osmium safety advice etc. CORRECT STORAGE

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Tom,

Wow. Really? I've put osmium waste in oil
everywhere I've been, or seen that where I've
gone. I've surprised to see putting osmium in oil
as "processing" and not "storage" (or some
similar synonym).
Mind, the simplest is just to do it, and state
it's part of the experimental protocol. The
osmium-in-oil (aka osmium dioxide bound to
unsaturated carbon-carbon bonds in an oily
polymer) is the waste. Not the osmium removed
from the specimens.

Phil

} First, let me strongly suggest you not store
} unreacted waste in Mason jars in your
} refrigerator. My experience is no single jar
} seal is sufficient to contain osmium vapor. I
} store my 2% osmium and osmium waste in my fume
} hood. My 2% osmium stock is in a tightly sealed
} orange-cap Schott bottle. The orange cap has
} gone completely black. The entire Schott bottle
} is kept inside a clear plastic polycarbonate
} jar. The polycarbonate has gone completely
} black. I have seen, in other labs in the past,
} refrigerators with blackened walls due to
} storage of osmium as you suggest you do. As a
} general rule, store your osmium in double
} containers and keep them in a fume hood.
}
} Secondly, it is not allowed at my University for
} an investigator to "process" hazardous waste
} without a written protocol that has been
} approved. I can't simply put my osmium waste in
} oil such as Debbie suggests. I think I once
} asked about that and was told not to do that. I
} don't know the rationale behind this denial.
}
}
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://www.biology.missouri.edu/faculty/phillips.html
} http://www.biotech.missouri.edu/mcc/
}
}
} -----Original Message-----
} X-from: David.Patton-at-uwe.ac.uk [mailto:David.Patton-at-uwe.ac.uk]
} Sent: Wednesday, November 10, 2010 9:44 AM
} To: Phillips, Thomas E.
} Subject: [Microscopy] Osmium safety advice etc. CORRECT STORAGE
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576


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From: richard.miron-at-zmk.unibe.ch
Date: Wed, 10 Nov 2010 10:29:14 -0600
Subject: [Microscopy] Aurion R-Gent SE-LM Silver Enhancement Reagents

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hello,

Has anyone modified their protocol or has successfully used aurion
silver enhancement reagent for SE to LM. So far we haven?t been able
to detect any silver enhancement but our gold labeling has worked in
TEM for LR White sections and the antibody is used successfully for
fluorescence and paraffin sections. Does anyone modify their protocol
to improve their results. Aurion doesn?t seem to know what?s wrong
and I?ve recently tried it with a second antibody.

Rick



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From: PhillipsT-at-missouri.edu
Date: Wed, 10 Nov 2010 10:37:13 -0600
Subject: [Microscopy] RE: Osmium safety advice etc. CORRECT STORAGE

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Years ago, a guy in charge of our hazard waste told me one rationale for not allowing any "processing". Corporations had to pay $XXX dollars to get rid of barrels of anything that contained what was classified as "hazardous". But barrels of very low 3H radioactivity were allowed to be poured down the drain with plenty of rinsing under federal guidelines. The scam that some industrial corporations were doing was adding a small amount of radioactivity to their waste stream. Those "hazardous" barrels were now under federal "radioisotope" waste restrictions and therefore could be poured down the drain at a significant cost savings. I believe this loophole has been plugged.

We also can't call anything in our lab waste nor can we label it as "waste". We only have "Used, unwanted chemicals" since "waste" implies it has to be handled with a special protocol and the "used, unwanted chemicals" folks don't want their hands tied.

The EHS folks at my university have some tedious rules but one thing they do deserves great credit and should be copied at all universities. They do a special pickup for any "unwanted chemicals" which includes any chemical that you use 1 mg in an experiment once or twice and don't need the remaining 199.8 gm in the original bottle. These chemicals are stored in their facility, listed on a website and available for any registered user on campus to go and get for free. Lots of chemicals don't go bad just because they are a year or ten old! In many ways this is no different than using an older bottle on your own shelf or borrowing from a neighboring lab. I can't tell you how often this has saved me money and allowed me to do a pilot experiment I wouldn't have tried if it meant ordering some $100 bottle. If your university doesn't have this, you should agitate for it. It saves money and reduces the toxic waste stream.


Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Wednesday, November 10, 2010 10:25 AM
To: Phillips, Thomas E.

Tom,

Wow. Really? I've put osmium waste in oil
everywhere I've been, or seen that where I've
gone. I've surprised to see putting osmium in oil
as "processing" and not "storage" (or some
similar synonym).
Mind, the simplest is just to do it, and state
it's part of the experimental protocol. The
osmium-in-oil (aka osmium dioxide bound to
unsaturated carbon-carbon bonds in an oily
polymer) is the waste. Not the osmium removed
from the specimens.

Phil

} First, let me strongly suggest you not store
} unreacted waste in Mason jars in your
} refrigerator. My experience is no single jar
} seal is sufficient to contain osmium vapor. I
} store my 2% osmium and osmium waste in my fume
} hood. My 2% osmium stock is in a tightly sealed
} orange-cap Schott bottle. The orange cap has
} gone completely black. The entire Schott bottle
} is kept inside a clear plastic polycarbonate
} jar. The polycarbonate has gone completely
} black. I have seen, in other labs in the past,
} refrigerators with blackened walls due to
} storage of osmium as you suggest you do. As a
} general rule, store your osmium in double
} containers and keep them in a fume hood.
}
} Secondly, it is not allowed at my University for
} an investigator to "process" hazardous waste
} without a written protocol that has been
} approved. I can't simply put my osmium waste in
} oil such as Debbie suggests. I think I once
} asked about that and was told not to do that. I
} don't know the rationale behind this denial.
}
}
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://www.biology.missouri.edu/faculty/phillips.html
} http://www.biotech.missouri.edu/mcc/
}
}
} -----Original Message-----
} X-from: David.Patton-at-uwe.ac.uk [mailto:David.Patton-at-uwe.ac.uk]
} Sent: Wednesday, November 10, 2010 9:44 AM
} To: Phillips, Thomas E.
} Subject: [Microscopy] Osmium safety advice etc. CORRECT STORAGE
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576


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From: ehaller-at-health.usf.edu
Date: Wed, 10 Nov 2010 10:39:24 -0600
Subject: [Microscopy] Re: Osmium safety advice etc.

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi, Dave,

I keep my Health and Safety people happy with plastic waste containers. I use empty ethanol bottles. I purchase 100% ethanol in pint bottles, then use them for my osmium disposal. I purchase 95% ethanol in gallon plastic bottles, then use these for my glutaraldehyde disposal. Plastic will not break if dropped, and is non-reactive to either chemical. Instead of oil to neutralize osmium, I use either alcohol or acetone, both of which turn osmium tetroxide into osmium black, which is inert. I start the waste bottle with 10-15mL of 100% ethanol in it, add my waste osmium, then put my lower percent dehydrating fluids (35%, 50%) in with my osmium. I end up with less volume to dispose of, and it still renders the osmium unreactive. You can test this by looking at the lid and walls of the ethanol bottle. They will turn black if the osmium is reactive, but remain white if the osmium is neutralized. My ethanol bottles are always white when I call Health and Safety to pick up my osmium waste. As a rule, I'm always working with 1-2mL of osmium per sample, so my waste volumes are not that much to begin with. I also collect my solid waste contaminated with either osmium or glutaraldehyde (pipets, tissues, vials) in a wide-mouthed plastic jar, seal it and have Health and Safety pick it up. These things stay reactive, so only open them in the fume hood. I store all of these containers in the hood until disposal.

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-cas.usf.edu
________________________________________
X-from: David.Patton-at-uwe.ac.uk [David.Patton-at-uwe.ac.uk]
Sent: Wednesday, November 10, 2010 10:51 AM
To: Haller, Edward

I would like to broaden the topic a little and request advice.

Osmium. I have been hording unreacted waste in Kilner/Mason jars in my fridge. I suppose that when it is reacted with oil then it is no longer volatile. Is this mixture in a polypropylene bottle with an ordinary screw lid a satisfactory container to pass on to a waste disposal operative?

Supplementary question: Phil - Kitty litter - I presume you mean the mineral type - how much do you use? Eg 10% of the bottle's volume?

Waste gluraraldehyde in buffer. This will probably still be volatile. What is a satisfactory container to pass on to a waste disposal operative?

Dave

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: 10 November 2010 14:38
To: David Patton

Kristen,

First, to echo other responders, don't worry
about teaching to undergrads. We have a
microscopy major within our Biology B.Sc. program
and routinely teach TEM and SEM to undergrads.
I've never found that undergrads are a big safety
problem. (Engineering grad students, now ...)
Teach proper techniques (not over-done panicky
Safety Committee Ohmygodosmium!) and they'll be
fine. Do emphasize that OsO4 is highly volatile
and *must* be used in a hood. Show them a
blackened OsO4 storage container and the inside
of your fixative 'frig. They'll get the point.
Keep a spill kit handy and show them how to use it.
OsO4 waste: as others have written, use vegetable
oil in a plastic container (not glass - glass
breaks, then you have a nasty mess), but find the
cheap highly polyunsaturated oil, like Canola.
The more unsaturated the oil, the more it binds
up OsO4. Also, add kitty litter to the waste
container. This keeps the oil from sloshing
around and increases the surface area of oil
available to bind OsO4.
And! Have a separate wide-mouth container for
solid OsO4 contaminated waste - transfer
pipettes, gloves, etc.
Emergency respirator - filtered is fine IF it's
the right filter. But mostly, have kitty litter
and oil handy to dump on any spill. But, if the
OsO4 is kept in the hood, then any spill should
be in the hood - preferably in a tray - so a
respirator really isn't necessary.

Re: samples. You can get away with using tissues
from a local vet or other people's research.
Unless you have an active animal-care committee
and animal research protocols. Then you'll likely
discover that you must have your own animal
tissue protocols even if you are using samples
taken from animals used in research or brought in
from off campus.
But.
If you use insects like crickets - used for
reptile food - or the cockroaches and spiders
hanging around your building, then you have no
issues with animal care committees, animal use
protocols (there are laws and regulations
involved here beyond your campus regs) and so on.
(Or isopods - pillbugs and sowbugs - or worms or
etc.) Fruitflies.
Plus, you get really neat specimens:
contracted/relaxed muscles, Malpighian tubules,
venom and silk glands, and lots more.
And there are lots of EM images in the literature
and the 15 volume "Microscopic Anatomy of
Invertebrates" published by Wiley-Liss and Miriam
Rothschild's beautiful "Insect Tissues via the
Flea".
Or, brine shrimp (Artemia) are easy to culture.
All the specimens for this would be free.

Note: just dumping testicles and ovaries into a
jar of Karnovsky's would be a good exercise in
poor preservation and why one minces tissue for
EM fixation.

Phil

} Hi Listers,
}
} I'm gearing up to teach introductory electron microscopy to undergraduate
} biology majors again this Spring and have hit a
} few safety road blocks on which
} I'd greatly appreciate your advice. Yes, undergrads in an EM lab is a big, red
} safety flag on it's own!
}
} First, I'm on my own with this, as there is no one else for miles who has any
} expertise with EM and we don't have a dedicated health and safety unit at my
} University. I am very spoiled in that I've
} always been an EM user with dedicated
} staff and safety personnel within reach. So, the first thing thatÝI need is a
} full face respirator in the event that someone
} drops Osmium because it's all up
} to me to clean it up. My first question is, do I need filtered air or supplied
} air? Any other suggestions?
}
} Secondly, I have a faint memory of there being a way to reduce the toxicity of
} Osmium waste by binding the osmium to oil. Am I hallucinating or does someone
} have some tips on doing this? We're trying to reduce the toxicity and save the
} University some cash in waste disposal fees because it will be cheaper to deal
} with less toxic waste.
}
}
} Now, a couple of other question related to the class. I know that I'm going to
} run into some trouble when I tell our budget
} keepers that I've got to order new
} supplies because 2 year old glut is not acceptable. EM is expensive! We have a
} heavy focus on field science, so the expense of this class comes as a shock.
} Does anyone have any insight as to the "shelf life" of unopened ampules of
} glutaraldehyde? Maybe this would be a better buy this year.
}
} Lastly, as a plant person, I'm struggling with
} how toÝget my students experience
} with animal tissueÝbecause I lost my colleague whoÝused to sacrificeÝrats for
} his research and lend me some tissue for my class. I do have a connection to a
} local vet and am wondering if anyone has any
} advice as to whether having him pop
} some of the tissues that he harvests from his
} patients into vials of Karnovsky's
} would be a good option. My students may get a
} lot of cat testicles and ovaries,
} but at least they'd get some animal tissue!
}
} Thanks in advance for any advice you can lend.
} I'm sure that I'll be in touch as
} I muddle my way through another semester of enlightening undergrads.
} -Kristen Lennon
}
} Kristen Lennon, Ph.D.
} Department of Biology
} Frostburg State University
} Frostburg, MD 21532
} kalennon-at-frostburg.edu

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576


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From: dsherman-at-purdue.edu
Date: Wed, 10 Nov 2010 10:43:03 -0600
Subject: [Microscopy] Re: Osmium safety advice etc. CORRECT STORAGE

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dave,
We use polypropylene as it is inert and won't break. We like the 1 liter
size as that is the most I want to accumulate before getting it out of the
lab. Large mouth screw cap works great as it is easy to pour into the
container with out spillage and easy to close. We can get these bottles
from campus stores and use them for all our waste products (fixatives, glass
vials from fixatives, resins, ETOH, acetone, etc)...each in a separate
container and appropriately labeled.
--
Debby


} From: "David.Patton-at-uwe.ac.uk" {David.Patton-at-uwe.ac.uk}
} Reply-To: "David.Patton-at-uwe.ac.uk" {David.Patton-at-uwe.ac.uk}
} Date: Wed, 10 Nov 2010 10:44:54 -0500
} To: Debby Sherman {dsherman-at-purdue.edu}
} Subject: [Microscopy] Osmium safety advice etc. CORRECT STORAGE
}
}
}
}
} ----------------------------------------------------------------------------
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}
} I would like to broaden the topic a little and request advice.
}
} Osmium. I have been hording unreacted waste in Kilner/Mason jars in my fridge.
} I suppose that when it is reacted with oil then it is no longer volatile. Is
} this mixture in a polypropylene bottle with an ordinary screw lid a
} satisfactory container to pass on to a waste disposal operative?
}
} Supplementary question: Phil - Kitty litter - I presume you mean the mineral
} type - how much do you use? Eg 10% of the bottle's volume?
}
} Waste gluraraldehyde in buffer. This will probably still be volatile. What
} is a satisfactory container to pass on to a waste disposal operative?
}
} Dave
}
} -----Original Message-----
} X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
} Sent: 10 November 2010 14:38
} To: David Patton
} Subject: [Microscopy] Re: Osmium safety advice etc.
}
}
}
}
} ----------------------------------------------------------------------------
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}
} Kristen,
}
} First, to echo other responders, don't worry
} about teaching to undergrads. We have a
} microscopy major within our Biology B.Sc. program
} and routinely teach TEM and SEM to undergrads.
} I've never found that undergrads are a big safety
} problem. (Engineering grad students, now ...)
} Teach proper techniques (not over-done panicky
} Safety Committee Ohmygodosmium!) and they'll be
} fine. Do emphasize that OsO4 is highly volatile
} and *must* be used in a hood. Show them a
} blackened OsO4 storage container and the inside
} of your fixative 'frig. They'll get the point.
} Keep a spill kit handy and show them how to use it.
} OsO4 waste: as others have written, use vegetable
} oil in a plastic container (not glass - glass
} breaks, then you have a nasty mess), but find the
} cheap highly polyunsaturated oil, like Canola.
} The more unsaturated the oil, the more it binds
} up OsO4. Also, add kitty litter to the waste
} container. This keeps the oil from sloshing
} around and increases the surface area of oil
} available to bind OsO4.
} And! Have a separate wide-mouth container for
} solid OsO4 contaminated waste - transfer
} pipettes, gloves, etc.
} Emergency respirator - filtered is fine IF it's
} the right filter. But mostly, have kitty litter
} and oil handy to dump on any spill. But, if the
} OsO4 is kept in the hood, then any spill should
} be in the hood - preferably in a tray - so a
} respirator really isn't necessary.
}
} Re: samples. You can get away with using tissues
} from a local vet or other people's research.
} Unless you have an active animal-care committee
} and animal research protocols. Then you'll likely
} discover that you must have your own animal
} tissue protocols even if you are using samples
} taken from animals used in research or brought in
} from off campus.
} But.
} If you use insects like crickets - used for
} reptile food - or the cockroaches and spiders
} hanging around your building, then you have no
} issues with animal care committees, animal use
} protocols (there are laws and regulations
} involved here beyond your campus regs) and so on.
} (Or isopods - pillbugs and sowbugs - or worms or
} etc.) Fruitflies.
} Plus, you get really neat specimens:
} contracted/relaxed muscles, Malpighian tubules,
} venom and silk glands, and lots more.
} And there are lots of EM images in the literature
} and the 15 volume "Microscopic Anatomy of
} Invertebrates" published by Wiley-Liss and Miriam
} Rothschild's beautiful "Insect Tissues via the
} Flea".
} Or, brine shrimp (Artemia) are easy to culture.
} All the specimens for this would be free.
}
} Note: just dumping testicles and ovaries into a
} jar of Karnovsky's would be a good exercise in
} poor preservation and why one minces tissue for
} EM fixation.
}
} Phil
}
} } Hi Listers,
} }
} } I'm gearing up to teach introductory electron microscopy to undergraduate
} } biology majors again this Spring and have hit a
} } few safety road blocks on which
} } I'd greatly appreciate your advice. Yes, undergrads in an EM lab is a big,
} } red
} } safety flag on it's own!
} }
} } First, I'm on my own with this, as there is no one else for miles who has any
} } expertise with EM and we don't have a dedicated health and safety unit at my
} } University. I am very spoiled in that I've
} } always been an EM user with dedicated
} } staff and safety personnel within reach. So, the first thing thatÝI need is a
} } full face respirator in the event that someone
} } drops Osmium because it's all up
} } to me to clean it up. My first question is, do I need filtered air or
} } supplied
} } air? Any other suggestions?
} }
} } Secondly, I have a faint memory of there being a way to reduce the toxicity
} } of
} } Osmium waste by binding the osmium to oil. Am I hallucinating or does someone
} } have some tips on doing this? We're trying to reduce the toxicity and save
} } the
} } University some cash in waste disposal fees because it will be cheaper to
} } deal
} } with less toxic waste.
} }
} }
} } Now, a couple of other question related to the class. I know that I'm going
} } to
} } run into some trouble when I tell our budget
} } keepers that I've got to order new
} } supplies because 2 year old glut is not acceptable. EM is expensive! We have
} } a
} } heavy focus on field science, so the expense of this class comes as a shock.
} } Does anyone have any insight as to the "shelf life" of unopened ampules of
} } glutaraldehyde? Maybe this would be a better buy this year.
} }
} } Lastly, as a plant person, I'm struggling with
} } how toÝget my students experience
} } with animal tissueÝbecause I lost my colleague whoÝused to sacrificeÝrats for
} } his research and lend me some tissue for my class. I do have a connection to
} } a
} } local vet and am wondering if anyone has any
} } advice as to whether having him pop
} } some of the tissues that he harvests from his
} } patients into vials of Karnovsky's
} } would be a good option. My students may get a
} } lot of cat testicles and ovaries,
} } but at least they'd get some animal tissue!
} }
} } Thanks in advance for any advice you can lend.
} } I'm sure that I'll be in touch as
} } I muddle my way through another semester of enlightening undergrads.
} } -Kristen Lennon
} }
} } Kristen Lennon, Ph.D.
} } Department of Biology
} } Frostburg State University
} } Frostburg, MD 21532
} } kalennon-at-frostburg.edu
}
} --
} Philip Oshel
} Microscopy Facility Supervisor
} Biology Department
} 024C Brooks Hall
} Central Michigan University
} Mt. Pleasant, MI 48859
} (989) 774-3576
}
}
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} 19, 36 -- Date: Wed, 10 Nov 2010 15:42:41 +0000
} 19, 36 -- From: David Patton {David.Patton-at-uwe.ac.uk}
} 19, 36 -- Subject: RE: [Microscopy] Re: Osmium safety advice etc. CORRECT
} STORAGE
} 19, 36 -- CONTAINERS FOR WASTE OSMIUM AND GLUTARALDEHYDE
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From: jkrupp-at-deltacollege.edu
Date: Wed, 10 Nov 2010 11:52:17 -0600
Subject: [Microscopy] Thick sections fall off

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Greetings

Some of my students have asked me to inquire about a problem they are having with thick plastic sections falling off their glass slides.

So, any advice or tricks of the trade? These are 1 um or so plastic sections, dried down in a drop of water onto a clean glass slide. The sections seem to slip off the slide during rinsing after toluidine blue staining. These are very good students who will appreciate any replies.

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu





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From: oshel1pe-at-cmich.edu
Date: Wed, 10 Nov 2010 12:08:41 -0600
Subject: [Microscopy] Re: Thick sections fall off

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Jon,

What temperature are they using to dry down the slides & for
staining? We use ('round about) 90 C. And *gentle* washing.

Phil

} Greetings
}
} Some of my students have asked me to inquire about a problem they
} are having with thick plastic sections falling off their glass
} slides.
}
} So, any advice or tricks of the trade? These are 1 um or so plastic
} sections, dried down in a drop of water onto a clean glass slide.
} The sections seem to slip off the slide during rinsing after
} toluidine blue staining. These are very good students who will
} appreciate any replies.
}
} Jon
}
} Jonathan Krupp
} Delta College
} 5151Pacific Ave.
} Stockton, CA 95207
} 209-954-5284
} jkrupp-at-deltacollege.edu

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: jsiegmund-at-7thwavelabs.com
Date: Wed, 10 Nov 2010 12:10:56 -0600
Subject: [Microscopy] Thick sections fall off

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html



Jon,
Are you using adhesive slides? If yes, try to leave the slides with
sections 20 min on the hotplate at around 58C, before staining.

Joe,

7thwaveLaboratories
Chesterfield, MO


-----Original Message-----
X-from: jkrupp-at-deltacollege.edu [mailto:jkrupp-at-deltacollege.edu]
Sent: Wednesday, November 10, 2010 12:01 PM
To: Joachim Siegmund

Greetings

Some of my students have asked me to inquire about a problem they are
having with thick plastic sections falling off their glass slides.

So, any advice or tricks of the trade? These are 1 um or so plastic
sections, dried down in a drop of water onto a clean glass slide. The
sections seem to slip off the slide during rinsing after toluidine blue
staining. These are very good students who will appreciate any replies.

Jon

Jonathan Krupp
Delta College
5151Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu





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From: W.Muss-at-salk.at
Date: Wed, 10 Nov 2010 12:14:14 -0600
Subject: [Microscopy] Re: Osmium safety advice etc. // CORRECT STORAGE & Fix GA

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Ed,
(dear all),

I am really delighted you pointed to the possibility (and your method) to reduce OsO4 to OsO2 (as it in fact will happen during dehydration of osmicated specimens too) by using ("absolute") EtOH, most preferably from the last dehydration steps one has to perform.

Re: normally one ends up with dehydration of spec's (e. g. at least 2 - 3 times EtOH absolute waterfree... yes, also you are right with acetone which for that purpose is suited as well).
I don't know what others do with those fractions of used EtOH or Acetone.... But:

I use those 3-2 fractions of nearly anhydrous EtOH to reduce the used osmium since 1985 in our lab (and have presented a poster on this -at- EMSA 50th Ann. Meeting Boston 1992, as well as several personal postings on the MSA Listserver) instead of using "corn oil" which obviousely might prevent successful (or at least simple) recovery of OsO2 if this is wanted.


Note: In the Listserver Archive, 2001 (search phrase: osmium, some 25 postings )
on Wed, 23 May 2001 Dr. Frederick Monson stated (besides other postings):

=========================================================================================================
} } found this on the NET, and I hope Dr. Kiernan doesn't mind.

Re: "Disposal" of Osmium Tetroxide "Waste"
X-from: "J. A. Kiernan" {jkiernan-at-julian.uwo.ca}
------------------------------------------------------------------------
On Thu, 23 Nov 2000, stephen asquith wrote:

{ disposal of this wonderful material but it still ... } ... containers full of osmium in alcohol or corn oil ... } ... what should be done with it then?

Osmium tetroxide is indeed wonderful stuff. Os is a rare element, so disposal of used solutions should consist of recycling, not dumping, even though osmium compounds are not considered environmentally hazardous (Smith et al., 1978 Trace Metal in the Environment, vol 4. Ann Arbor Science Publishers). The colourless soluble toxic tetroxide is rapidly reduced by almost any kind of dirt to a black, insoluble dioxide, usually in a colloidal form that's readily dispersed by moving water if it isn't firmly stuck to the solid organic matter that brought about the reduction.
If OsO4 slops are collected in alcohol, the Osmium (now in the form of crude, harmless, insoluble osmium dioxide) can be reoxidized, purified, re-reduced to pure OsO2 and stored.

OsO2 is easily re-oxidized to give a buffered solution of osmium tetroxide (2% or less).
See J Microsc 113:77-82 (1978); the procedure does involve certain hazards, so it must be done carefully. Recovered OsO4 can also be used to make OSMETH, which is a beautiful golden crystalline solid that contains osmium tetroxide complexed with methenamine (= hexamethylene tetramine or hexamine (Hanker et al., 1976 Histochemistry 49:263-291). It costs next to nothing to make your own osmeth from recycled OsO4, but osmeth is very expensive to buy. Osmeth does not emit osmic fumes. When it is dissolved (in DMF followed by dilution in an aqueous buffer) it becomes a dilute (0.25%) working osmium tetroxide solution. I can vouch for the excellence of home-made osmeth for post-osmication (for EM). It may also be OK as a primary fixative or for LM staining, but I haven't encountered (personally, by anecdote or in the literature) any use of osmeth other than for postosmication. Perhaps someone reading this message will put me right on this. Osmium tetroxide collected into vegetable oil could not be recycled by the simple method cited above, and the recovery methods used by chemists (which use apparatus etc not found in histology labs) would be made more difficult by the presence of the oil. John A. Kiernan, Department of Anatomy & Cell Biology, The University of Western Ontario, LONDON, Canada N6A 5C1

Frederick C. Monson, PhD
West Chester University of Pennsylvania
Center for Advanced Scientific Imaging (CASI)
Schmucker II Science Center (Room: SS024(Basement))
South Church Street
West Chester, PA, 19383
MailDrop: Department of Geology/Astronomy
Phone: 610-738-0437
Fax: 610-436-3036
email: fmonson-at-wcupa.edu
Please call before visiting.
=====================================================================================================

Concerning disposal of Glutaraldehyde after prior deactivation see: eg.
http://www.dow.com/PublishedLiterature/dh_00de/0901b803800de37f.pdf?filepath=biocides/pdfs/noreg/253-01443.pdf&fromPage=GetDoc
, which mentions chemical deactivation via alkalinization or reducing processes like use of sodium bisulfite, sodium hydroxide (also find hints on disposal).
Smaller amounts and concentrations used for EM-prep's also IMO directly after fixation step(s) can be deactivated by overlaying hydrous NH4Cl (ammonium chloride, 0.05 - 1 M), practically spoken: if you decant or suck off fixative into a washing bottle and use one washing buffer containing 50 mM Ammonium chloride (recipe ROTH et al 1981; similar effects like use of Na-borohydride, glycine or lysine) you directly could deactivate at least most of the unreacted aldehydes.

Any practical advice with OsO4-Ethanol reduction, further storage/disposal and /or recovery I should be happy to tell on request (knowing that there might be differences in legisalation USA-Europe),

cordially yours,


Wolfgang MUSS PhD
Member of MSA since 1996
EM-Lab, Pathology SALK-LKH(Gen.Hosp.) & PMU
SALZBURG, AUSTRIA





} -----Ursprüngliche Nachricht-----
} Von: ehaller-at-health.usf.edu [mailto:ehaller-at-health.usf.edu]
} Gesendet: Mittwoch, 10. November 2010 17:43
} An: Muß Wolfgang
} Betreff: [Microscopy] Re: Osmium safety advice etc. // CORRECT STORAGE
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi, Dave,
}
} I keep my Health and Safety people happy with plastic waste
} containers.
} I use empty ethanol bottles. I purchase 100% ethanol in pint bottles,
} then use them for my osmium disposal.
} I purchase 95% ethanol in gallon plastic bottles, then use these for my
} glutaraldehyde disposal.
}
} Plastic will not break if dropped, and is non-reactive to either
} chemical.
} Instead of oil to neutralize osmium, I use either alcohol or acetone,
} both of which turn osmium tetroxide into osmium black, which is inert.
}
} I start the waste bottle with 10-15mL of 100% ethanol in it, add my
} waste osmium, then put my lower percent dehydrating fluids (35%, 50%)
} in with my osmium.
} I end up with less volume to dispose of, and it still renders the
} osmium unreactive. You can test this by looking at the lid and walls of
} the ethanol bottle.
} They will turn black if the osmium is reactive, but remain white if the
} osmium is neutralized. My ethanol bottles are always white when I call
} Health and Safety to pick up my osmium waste. As a rule, I'm always
} working with 1-2mL of osmium per sample, so my waste volumes are not
} that much to begin with.
}
} I also collect my solid waste contaminated with either osmium or
} glutaraldehyde (pipets, tissues, vials) in a wide-mouthed plastic jar,
} seal it and have Health and Safety pick it up. These things stay
} reactive, so only open them in the fume hood. I store all of these
} containers in the hood until disposal.
}
} Ed
}
} Edward Haller, Lab Manager
} University of South Florida
} Integrative Biology Department
} Electron Microscopy Core
} SCA 110
} 4202 East Fowler Avenue
} Tampa, FL 33620
} 813-974-2676
} ehaller-at-cas.usf.edu
} ________________________________________
} X-from: David.Patton-at-uwe.ac.uk [David.Patton-at-uwe.ac.uk]
} Sent: Wednesday, November 10, 2010 10:51 AM
} To: Haller, Edward
} Subject: [Microscopy] Osmium safety advice etc. CORRECT STORAGE
}
} -----------------------------------------------------------------------
} -----
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} -----------------------------------------------------------------------
} -----
}
} I would like to broaden the topic a little and request advice.
}
} Osmium. I have been hording unreacted waste in Kilner/Mason jars in my
} fridge. I suppose that when it is reacted with oil then it is no longer
} volatile. Is this mixture in a polypropylene bottle with an ordinary
} screw lid a satisfactory container to pass on to a waste disposal
} operative?
}
} Supplementary question: Phil - Kitty litter - I presume you mean the
} mineral type - how much do you use? Eg 10% of the bottle's volume?
}
} Waste gluraraldehyde in buffer. This will probably still be volatile.
} What is a satisfactory container to pass on to a waste disposal
} operative?
}
} Dave
}
} -----Original Message-----
} X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
} Sent: 10 November 2010 14:38
} To: David Patton
} Subject: [Microscopy] Re: Osmium safety advice etc.
}
}
}
}
} -----------------------------------------------------------------------
} -----
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} -----------------------------------------------------------------------
} -----
}
} Kristen,
}
} First, to echo other responders, don't worry
} about teaching to undergrads. We have a
} microscopy major within our Biology B.Sc. program
} and routinely teach TEM and SEM to undergrads.
} I've never found that undergrads are a big safety
} problem. (Engineering grad students, now ...)
} Teach proper techniques (not over-done panicky
} Safety Committee Ohmygodosmium!) and they'll be
} fine. Do emphasize that OsO4 is highly volatile
} and *must* be used in a hood. Show them a
} blackened OsO4 storage container and the inside
} of your fixative 'frig. They'll get the point.
} Keep a spill kit handy and show them how to use it.
} OsO4 waste: as others have written, use vegetable
} oil in a plastic container (not glass - glass
} breaks, then you have a nasty mess), but find the
} cheap highly polyunsaturated oil, like Canola.
} The more unsaturated the oil, the more it binds
} up OsO4. Also, add kitty litter to the waste
} container. This keeps the oil from sloshing
} around and increases the surface area of oil
} available to bind OsO4.
} And! Have a separate wide-mouth container for
} solid OsO4 contaminated waste - transfer
} pipettes, gloves, etc.
} Emergency respirator - filtered is fine IF it's
} the right filter. But mostly, have kitty litter
} and oil handy to dump on any spill. But, if the
} OsO4 is kept in the hood, then any spill should
} be in the hood - preferably in a tray - so a
} respirator really isn't necessary.
}
} Re: samples. You can get away with using tissues
} from a local vet or other people's research.
} Unless you have an active animal-care committee
} and animal research protocols. Then you'll likely
} discover that you must have your own animal
} tissue protocols even if you are using samples
} taken from animals used in research or brought in
} from off campus.
} But.
} If you use insects like crickets - used for
} reptile food - or the cockroaches and spiders
} hanging around your building, then you have no
} issues with animal care committees, animal use
} protocols (there are laws and regulations
} involved here beyond your campus regs) and so on.
} (Or isopods - pillbugs and sowbugs - or worms or
} etc.) Fruitflies.
} Plus, you get really neat specimens:
} contracted/relaxed muscles, Malpighian tubules,
} venom and silk glands, and lots more.
} And there are lots of EM images in the literature
} and the 15 volume "Microscopic Anatomy of
} Invertebrates" published by Wiley-Liss and Miriam
} Rothschild's beautiful "Insect Tissues via the
} Flea".
} Or, brine shrimp (Artemia) are easy to culture.
} All the specimens for this would be free.
}
} Note: just dumping testicles and ovaries into a
} jar of Karnovsky's would be a good exercise in
} poor preservation and why one minces tissue for
} EM fixation.
}
} Phil
}
} } Hi Listers,
} }
} } I'm gearing up to teach introductory electron microscopy to
} undergraduate
} } biology majors again this Spring and have hit a
} } few safety road blocks on which
} } I'd greatly appreciate your advice. Yes, undergrads in an EM lab is a
} big, red
} } safety flag on it's own!
} }
} } First, I'm on my own with this, as there is no one else for miles who
} has any
} } expertise with EM and we don't have a dedicated health and safety unit
} at my
} } University. I am very spoiled in that I've
} } always been an EM user with dedicated
} } staff and safety personnel within reach. So, the first thing thatÝI
} need is a
} } full face respirator in the event that someone
} } drops Osmium because it's all up
} } to me to clean it up. My first question is, do I need filtered air or
} supplied
} } air? Any other suggestions?
} }
} } Secondly, I have a faint memory of there being a way to reduce the
} toxicity of
} } Osmium waste by binding the osmium to oil. Am I hallucinating or does
} someone
} } have some tips on doing this? We're trying to reduce the toxicity and
} save the
} } University some cash in waste disposal fees because it will be cheaper
} to deal
} } with less toxic waste.
} }
} }
} } Now, a couple of other question related to the class. I know that I'm
} going to
} } run into some trouble when I tell our budget
} } keepers that I've got to order new
} } supplies because 2 year old glut is not acceptable. EM is expensive!
} We have a
} } heavy focus on field science, so the expense of this class comes as a
} shock.
} } Does anyone have any insight as to the "shelf life" of unopened
} ampules of
} } glutaraldehyde? Maybe this would be a better buy this year.
} }
} } Lastly, as a plant person, I'm struggling with
} } how toÝget my students experience
} } with animal tissueÝbecause I lost my colleague whoÝused to
} sacrificeÝrats for
} } his research and lend me some tissue for my class. I do have a
} connection to a
} } local vet and am wondering if anyone has any
} } advice as to whether having him pop
} } some of the tissues that he harvests from his
} } patients into vials of Karnovsky's
} } would be a good option. My students may get a
} } lot of cat testicles and ovaries,
} } but at least they'd get some animal tissue!
} }
} } Thanks in advance for any advice you can lend.
} } I'm sure that I'll be in touch as
} } I muddle my way through another semester of enlightening undergrads.
} } -Kristen Lennon
} }
} } Kristen Lennon, Ph.D.
} } Department of Biology
} } Frostburg State University
} } Frostburg, MD 21532
} } kalennon-at-frostburg.edu
}
} --
} Philip Oshel
} Microscopy Facility Supervisor
} Biology Department
} 024C Brooks Hall
} Central Michigan University
} Mt. Pleasant, MI 48859
} (989) 774-3576
}
}
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From: W.Muss-at-salk.at
Date: Wed, 10 Nov 2010 12:30:37 -0600
Subject: [Microscopy] Re: Thick sections fall off

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html


Dear Jon,

hope your students will be delighted to know that - at least using Epon or Epon-Substitutes (like glycidether 100, Embed or also LADD LX-112 expoxy resins) - I did have no problems staining large (up to 4 x 5 mm ) 1µm sections with a multi-step polychromatic staining sequence (at least one step 8 min -at- 80 degrees C) if the sections were left on a hotplate -at-100-120 degrees C for at least 5 min.

Certainly there could be differences in treating (heating) sections on to the glass slides for other resin types, critical in my opinion too perhaps is age and cleanliness of slides (mine before use are batch-treated in diluted HCl-EtOH solution, rinsed thoroughly in A. dest and dried in a dustfree location before use).

Heating sections to such high temperatures before staining IMO have not much adverse effects in terms of staining quality or intensity.


Best regards and good luck,

Wolfgang MUSS
EM-Lab, Pathology
SALK-LKH (Gen.Hosp.) & PMU
SALZBURG, Austria



} -----Ursprüngliche Nachricht-----
} Von: jsiegmund-at-7thwavelabs.com [mailto:jsiegmund-at-7thwavelabs.com]
} Gesendet: Mittwoch, 10. November 2010 19:13
} An: Muß Wolfgang
} Betreff: [Microscopy] RE: Thick sections fall off
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} Jon,
} Are you using adhesive slides? If yes, try to leave the slides with
} sections 20 min on the hotplate at around 58C, before staining.
}
} Joe,
}
} 7thwaveLaboratories
} Chesterfield, MO
}
}
} -----Original Message-----
} X-from: jkrupp-at-deltacollege.edu [mailto:jkrupp-at-deltacollege.edu]
} Sent: Wednesday, November 10, 2010 12:01 PM
} To: Joachim Siegmund
} Subject: [Microscopy] Thick sections fall off
}
}
}
}
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}
} Greetings
}
} Some of my students have asked me to inquire about a problem they are
} having with thick plastic sections falling off their glass slides.
}
} So, any advice or tricks of the trade? These are 1 µm or so plastic
} sections, dried down in a drop of water onto a clean glass slide. The
} sections seem to slip off the slide during rinsing after toluidine blue
} staining. These are very good students who will appreciate any replies.
}
} Jon
}
} Jonathan Krupp
} Delta College
} 5151Pacific Ave.
} Stockton, CA 95207
} 209-954-5284
} jkrupp-at-deltacollege.edu
}
}
}
}
}
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From: kderr-at-nysbc.org
Date: Wed, 10 Nov 2010 12:30:57 -0600
Subject: [Microscopy] Re: Osmium safety advice etc. CORRECT STORAGE?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I work at a private facility without a safety office to drop my waste off
at. Waste disposal comes directly out of my budget. I found neutralizing
osmium with oil = 3 times the waste volume = $$$$(not to mention the
environmental impact). Tannic acid is a mordant for osmium = I neutralize
osmium with tannic acid. I allow this to dry out in the fume hood reducing
the waste volume. My fume hood walls, ceiling and dampers are still white
which I take as indication that this is working as planned. I have never
heard on anyone else neutralizing osmium in this way. Does anyone see
pitfalls to this method that I may have missed?

I am intrigued about using ethanol and/or acetone, what is the basis for
this? Anyone use a similar method?

Cheers,
KD Derr
Technical Assistant
New York Structural Biology Center





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Before we take our osmium waste to our Safety Office we store it with
sawdust and oil in a plastic bottle (for the same purpose as cat litter).
About 50-75% of the 1 litre bottle is filled with sawdust, and enough
vegetable oil is poured in to generously cover the sawdust (which compacts
down a bit)- no real reason why we use these quantities, but they seem
sensible amounts.

We also have a dedicated bin for osmium-contaminated gloves, disposable
pipettes and benchcoat; after they have been left for at least 12 hours in
the hood for the osmium to evaporate. I think this bin is emptied into the
normal lab waste bins, but it is contained within black bags and treated
more sensitively to prevent exposure of the staff emptying bins.


Ben


--
Imaging Technician
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271866
{http://mrcanu.pharm.ox.ac.uk/}




On 10/11/2010 15:51, "David.Patton-at-uwe.ac.uk" {David.Patton-at-uwe.ac.uk}
wrote:


} I would like to broaden the topic a little and request advice.
}
} Osmium. I have been hording unreacted waste in Kilner/Mason jars in my
} fridge. I suppose that when it is reacted with oil then it is no longer
} volatile. Is this mixture in a polypropylene bottle with an ordinary
} screw lid a satisfactory container to pass on to a waste disposal
} operative?
}
} Supplementary question: Phil - Kitty litter - I presume you mean the
} mineral type - how much do you use? Eg 10% of the bottle's volume?
}
} Waste gluraraldehyde in buffer. This will probably still be volatile.
} What is a satisfactory container to pass on to a waste disposal operative?
}
} Dave



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From: Shea.Miller-at-AGR.GC.CA
Date: Wed, 10 Nov 2010 12:36:27 -0600
Subject: [Microscopy] GMA resin

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I have been making my own GMA for years...

My "standard" recipe (to make ~100 mL:

2-hydroxyethyl methacrylate (glycol methacrylate) 95 mL
Carbowax 400 (polyethylene glycol) 5 mL
Benzoyl peroxide 0.5%

Mix on stirrer (gently.... Don't want to incorporate oxygen) until bp dissolves. You can store it in the freezer if you don't use the whole thing at once.
I usually polymerize at low temp (40-45 degrees) for about 48 hrs, then raise the temp to about 55 for a further 24 hrs to finish it off. Supposedly, you can manipulate the relative hardness by changing the amount of Polyethylene glycol 400, but I have never experimented with that.

Original reference: Feder, N. and O'Brien, T.P. (1968) Plant Microtechnique: Some Principles and New Methods. American Journal of Botany 55 (1): 123-142



Dr. S. Shea Miller
ECORC | CRECO
Agriculture and Agri-Food Canada | Agriculture et Agroalimentaire Canada
960 Carling Avenue | 960, avenue Carling
Ottawa, ON K1A 0C6
E-mail Address / Adresse courriel shea.miller-at-agr.gc.ca
Telephone | Téléphone 613-759-1760
Facsimile | Télécopieur 613-759-1701
Teletypewriter | Téléimprimeur 613-773-2600
Government of Canada | Gouvernement du Canada



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From: ehaller-at-health.usf.edu
Date: Wed, 10 Nov 2010 12:59:22 -0600
Subject: [Microscopy] Re: Osmium safety advice etc. // CORRECT STORAGE & Fix GA

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi, Wolfgang!

Thank you for saving that valuble information, and refreshing our memories! It was good to have the information in one place. I discovered the osmium and alcohol or acetone trick independantly decades ago and have been using it since. I didn't like the volume of corn oil waste that was generated, and knew that our tissue was non-reactive after processing for TEM or SEM, so I applied the same logic to osmium waste. Thank you for the references to back up the discovery! I'm glad that it was published. I also appreciate the information about neutralizing glutaraldehyde.

Sincerely,

Ed Haller

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-cas.usf.edu
________________________________________
X-from: W.Muss-at-salk.at [W.Muss-at-salk.at]
Sent: Wednesday, November 10, 2010 1:24 PM
To: Haller, Edward

Dear Ed,
(dear all),

I am really delighted you pointed to the possibility (and your method) to reduce OsO4 to OsO2 (as it in fact will happen during dehydration of osmicated specimens too) by using ("absolute") EtOH, most preferably from the last dehydration steps one has to perform.

Re: normally one ends up with dehydration of spec's (e. g. at least 2 - 3 times EtOH absolute waterfree... yes, also you are right with acetone which for that purpose is suited as well).
I don't know what others do with those fractions of used EtOH or Acetone.... But:

I use those 3-2 fractions of nearly anhydrous EtOH to reduce the used osmium since 1985 in our lab (and have presented a poster on this -at- EMSA 50th Ann. Meeting Boston 1992, as well as several personal postings on the MSA Listserver) instead of using "corn oil" which obviousely might prevent successful (or at least simple) recovery of OsO2 if this is wanted.


Note: In the Listserver Archive, 2001 (search phrase: osmium, some 25 postings )
on Wed, 23 May 2001 Dr. Frederick Monson stated (besides other postings):

=========================================================================================================
} } found this on the NET, and I hope Dr. Kiernan doesn't mind.

Re: "Disposal" of Osmium Tetroxide "Waste"
X-from: "J. A. Kiernan" {jkiernan-at-julian.uwo.ca}
------------------------------------------------------------------------
On Thu, 23 Nov 2000, stephen asquith wrote:

{ disposal of this wonderful material but it still ... } ... containers full of osmium in alcohol or corn oil ... } ... what should be done with it then?

Osmium tetroxide is indeed wonderful stuff. Os is a rare element, so disposal of used solutions should consist of recycling, not dumping, even though osmium compounds are not considered environmentally hazardous (Smith et al., 1978 Trace Metal in the Environment, vol 4. Ann Arbor Science Publishers). The colourless soluble toxic tetroxide is rapidly reduced by almost any kind of dirt to a black, insoluble dioxide, usually in a colloidal form that's readily dispersed by moving water if it isn't firmly stuck to the solid organic matter that brought about the reduction.
If OsO4 slops are collected in alcohol, the Osmium (now in the form of crude, harmless, insoluble osmium dioxide) can be reoxidized, purified, re-reduced to pure OsO2 and stored.

OsO2 is easily re-oxidized to give a buffered solution of osmium tetroxide (2% or less).
See J Microsc 113:77-82 (1978); the procedure does involve certain hazards, so it must be done carefully. Recovered OsO4 can also be used to make OSMETH, which is a beautiful golden crystalline solid that contains osmium tetroxide complexed with methenamine (= hexamethylene tetramine or hexamine (Hanker et al., 1976 Histochemistry 49:263-291). It costs next to nothing to make your own osmeth from recycled OsO4, but osmeth is very expensive to buy. Osmeth does not emit osmic fumes. When it is dissolved (in DMF followed by dilution in an aqueous buffer) it becomes a dilute (0.25%) working osmium tetroxide solution. I can vouch for the excellence of home-made osmeth for post-osmication (for EM). It may also be OK as a primary fixative or for LM staining, but I haven't encountered (personally, by anecdote or in the literature) any use of osmeth other than for postosmication. Perhaps someone reading this message will put me right on this. Osmium tetroxide collected into vegetable!

oil could not be recycled by the simple method cited above, and the recovery methods used by chemists (which use apparatus etc not found in histology labs) would be made more difficult by the presence of the oil. John A. Kiernan, Department of Anatomy & Cell Biology, The University of Western Ontario, LONDON, Canada N6A 5C1

Frederick C. Monson, PhD
West Chester University of Pennsylvania
Center for Advanced Scientific Imaging (CASI)
Schmucker II Science Center (Room: SS024(Basement))
South Church Street
West Chester, PA, 19383
MailDrop: Department of Geology/Astronomy
Phone: 610-738-0437
Fax: 610-436-3036
email: fmonson-at-wcupa.edu
Please call before visiting.
=====================================================================================================

Concerning disposal of Glutaraldehyde after prior deactivation see: eg.
http://www.dow.com/PublishedLiterature/dh_00de/0901b803800de37f.pdf?filepath=biocides/pdfs/noreg/253-01443.pdf&fromPage=GetDoc
, which mentions chemical deactivation via alkalinization or reducing processes like use of sodium bisulfite, sodium hydroxide (also find hints on disposal).
Smaller amounts and concentrations used for EM-prep's also IMO directly after fixation step(s) can be deactivated by overlaying hydrous NH4Cl (ammonium chloride, 0.05 - 1 M), practically spoken: if you decant or suck off fixative into a washing bottle and use one washing buffer containing 50 mM Ammonium chloride (recipe ROTH et al 1981; similar effects like use of Na-borohydride, glycine or lysine) you directly could deactivate at least most of the unreacted aldehydes.

Any practical advice with OsO4-Ethanol reduction, further storage/disposal and /or recovery I should be happy to tell on request (knowing that there might be differences in legisalation USA-Europe),

cordially yours,


Wolfgang MUSS PhD
Member of MSA since 1996
EM-Lab, Pathology SALK-LKH(Gen.Hosp.) & PMU
SALZBURG, AUSTRIA





} -----Ursprüngliche Nachricht-----
} Von: ehaller-at-health.usf.edu [mailto:ehaller-at-health.usf.edu]
} Gesendet: Mittwoch, 10. November 2010 17:43
} An: Muß Wolfgang
} Betreff: [Microscopy] Re: Osmium safety advice etc. // CORRECT STORAGE
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} ----------------------------------------------------------------------------
}
} Hi, Dave,
}
} I keep my Health and Safety people happy with plastic waste
} containers.
} I use empty ethanol bottles. I purchase 100% ethanol in pint bottles,
} then use them for my osmium disposal.
} I purchase 95% ethanol in gallon plastic bottles, then use these for my
} glutaraldehyde disposal.
}
} Plastic will not break if dropped, and is non-reactive to either
} chemical.
} Instead of oil to neutralize osmium, I use either alcohol or acetone,
} both of which turn osmium tetroxide into osmium black, which is inert.
}
} I start the waste bottle with 10-15mL of 100% ethanol in it, add my
} waste osmium, then put my lower percent dehydrating fluids (35%, 50%)
} in with my osmium.
} I end up with less volume to dispose of, and it still renders the
} osmium unreactive. You can test this by looking at the lid and walls of
} the ethanol bottle.
} They will turn black if the osmium is reactive, but remain white if the
} osmium is neutralized. My ethanol bottles are always white when I call
} Health and Safety to pick up my osmium waste. As a rule, I'm always
} working with 1-2mL of osmium per sample, so my waste volumes are not
} that much to begin with.
}
} I also collect my solid waste contaminated with either osmium or
} glutaraldehyde (pipets, tissues, vials) in a wide-mouthed plastic jar,
} seal it and have Health and Safety pick it up. These things stay
} reactive, so only open them in the fume hood. I store all of these
} containers in the hood until disposal.
}
} Ed
}
} Edward Haller, Lab Manager
} University of South Florida
} Integrative Biology Department
} Electron Microscopy Core
} SCA 110
} 4202 East Fowler Avenue
} Tampa, FL 33620
} 813-974-2676
} ehaller-at-cas.usf.edu
} ________________________________________
} X-from: David.Patton-at-uwe.ac.uk [David.Patton-at-uwe.ac.uk]
} Sent: Wednesday, November 10, 2010 10:51 AM
} To: Haller, Edward
} Subject: [Microscopy] Osmium safety advice etc. CORRECT STORAGE
}
} -----------------------------------------------------------------------
} -----
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} -----
}
} I would like to broaden the topic a little and request advice.
}
} Osmium. I have been hording unreacted waste in Kilner/Mason jars in my
} fridge. I suppose that when it is reacted with oil then it is no longer
} volatile. Is this mixture in a polypropylene bottle with an ordinary
} screw lid a satisfactory container to pass on to a waste disposal
} operative?
}
} Supplementary question: Phil - Kitty litter - I presume you mean the
} mineral type - how much do you use? Eg 10% of the bottle's volume?
}
} Waste gluraraldehyde in buffer. This will probably still be volatile.
} What is a satisfactory container to pass on to a waste disposal
} operative?
}
} Dave
}
} -----Original Message-----
} X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
} Sent: 10 November 2010 14:38
} To: David Patton
} Subject: [Microscopy] Re: Osmium safety advice etc.
}
}
}
}
} -----------------------------------------------------------------------
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}
} Kristen,
}
} First, to echo other responders, don't worry
} about teaching to undergrads. We have a
} microscopy major within our Biology B.Sc. program
} and routinely teach TEM and SEM to undergrads.
} I've never found that undergrads are a big safety
} problem. (Engineering grad students, now ...)
} Teach proper techniques (not over-done panicky
} Safety Committee Ohmygodosmium!) and they'll be
} fine. Do emphasize that OsO4 is highly volatile
} and *must* be used in a hood. Show them a
} blackened OsO4 storage container and the inside
} of your fixative 'frig. They'll get the point.
} Keep a spill kit handy and show them how to use it.
} OsO4 waste: as others have written, use vegetable
} oil in a plastic container (not glass - glass
} breaks, then you have a nasty mess), but find the
} cheap highly polyunsaturated oil, like Canola.
} The more unsaturated the oil, the more it binds
} up OsO4. Also, add kitty litter to the waste
} container. This keeps the oil from sloshing
} around and increases the surface area of oil
} available to bind OsO4.
} And! Have a separate wide-mouth container for
} solid OsO4 contaminated waste - transfer
} pipettes, gloves, etc.
} Emergency respirator - filtered is fine IF it's
} the right filter. But mostly, have kitty litter
} and oil handy to dump on any spill. But, if the
} OsO4 is kept in the hood, then any spill should
} be in the hood - preferably in a tray - so a
} respirator really isn't necessary.
}
} Re: samples. You can get away with using tissues
} from a local vet or other people's research.
} Unless you have an active animal-care committee
} and animal research protocols. Then you'll likely
} discover that you must have your own animal
} tissue protocols even if you are using samples
} taken from animals used in research or brought in
} from off campus.
} But.
} If you use insects like crickets - used for
} reptile food - or the cockroaches and spiders
} hanging around your building, then you have no
} issues with animal care committees, animal use
} protocols (there are laws and regulations
} involved here beyond your campus regs) and so on.
} (Or isopods - pillbugs and sowbugs - or worms or
} etc.) Fruitflies.
} Plus, you get really neat specimens:
} contracted/relaxed muscles, Malpighian tubules,
} venom and silk glands, and lots more.
} And there are lots of EM images in the literature
} and the 15 volume "Microscopic Anatomy of
} Invertebrates" published by Wiley-Liss and Miriam
} Rothschild's beautiful "Insect Tissues via the
} Flea".
} Or, brine shrimp (Artemia) are easy to culture.
} All the specimens for this would be free.
}
} Note: just dumping testicles and ovaries into a
} jar of Karnovsky's would be a good exercise in
} poor preservation and why one minces tissue for
} EM fixation.
}
} Phil
}
} } Hi Listers,
} }
} } I'm gearing up to teach introductory electron microscopy to
} undergraduate
} } biology majors again this Spring and have hit a
} } few safety road blocks on which
} } I'd greatly appreciate your advice. Yes, undergrads in an EM lab is a
} big, red
} } safety flag on it's own!
} }
} } First, I'm on my own with this, as there is no one else for miles who
} has any
} } expertise with EM and we don't have a dedicated health and safety unit
} at my
} } University. I am very spoiled in that I've
} } always been an EM user with dedicated
} } staff and safety personnel within reach. So, the first thing thatÝI
} need is a
} } full face respirator in the event that someone
} } drops Osmium because it's all up
} } to me to clean it up. My first question is, do I need filtered air or
} supplied
} } air? Any other suggestions?
} }
} } Secondly, I have a faint memory of there being a way to reduce the
} toxicity of
} } Osmium waste by binding the osmium to oil. Am I hallucinating or does
} someone
} } have some tips on doing this? We're trying to reduce the toxicity and
} save the
} } University some cash in waste disposal fees because it will be cheaper
} to deal
} } with less toxic waste.
} }
} }
} } Now, a couple of other question related to the class. I know that I'm
} going to
} } run into some trouble when I tell our budget
} } keepers that I've got to order new
} } supplies because 2 year old glut is not acceptable. EM is expensive!
} We have a
} } heavy focus on field science, so the expense of this class comes as a
} shock.
} } Does anyone have any insight as to the "shelf life" of unopened
} ampules of
} } glutaraldehyde? Maybe this would be a better buy this year.
} }
} } Lastly, as a plant person, I'm struggling with
} } how toÝget my students experience
} } with animal tissueÝbecause I lost my colleague whoÝused to
} sacrificeÝrats for
} } his research and lend me some tissue for my class. I do have a
} connection to a
} } local vet and am wondering if anyone has any
} } advice as to whether having him pop
} } some of the tissues that he harvests from his
} } patients into vials of Karnovsky's
} } would be a good option. My students may get a
} } lot of cat testicles and ovaries,
} } but at least they'd get some animal tissue!
} }
} } Thanks in advance for any advice you can lend.
} } I'm sure that I'll be in touch as
} } I muddle my way through another semester of enlightening undergrads.
} } -Kristen Lennon
} }
} } Kristen Lennon, Ph.D.
} } Department of Biology
} } Frostburg State University
} } Frostburg, MD 21532
} } kalennon-at-frostburg.edu
}
} --
} Philip Oshel
} Microscopy Facility Supervisor
} Biology Department
} 024C Brooks Hall
} Central Michigan University
} Mt. Pleasant, MI 48859
} (989) 774-3576
}
}
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From: baskin-at-bio.umass.edu
Date: Wed, 10 Nov 2010 13:29:13 -0600
Subject: [Microscopy] GMA resin --- and technovit

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Colleagues,
The original poster mentioned that
Technovit was a kind of GMA. I never knew that.
Sorry to be asking about "trade secrets" but does
anyone know how Technovit compares to the nice
and easy recipe from Shea Miller below??

Thanks,
Tobias

}
}
} I have been making my own GMA for years...
}
} My "standard" recipe (to make ~100 mL:
}
} 2-hydroxyethyl methacrylate (glycol methacrylate) 95 mL
} Carbowax 400 (polyethylene glycol) 5 mL
} Benzoyl peroxide
} 0.5%
}
} Mix on stirrer (gently.... Don't want to
} incorporate oxygen) until bp dissolves. You can
} store it in the freezer if you don't use the
} whole thing at once.
} I usually polymerize at low temp (40-45 degrees)
} for about 48 hrs, then raise the temp to about
} 55 for a further 24 hrs to finish it off.
} Supposedly, you can manipulate the relative
} hardness by changing the amount of Polyethylene
} glycol 400, but I have never experimented with
} that.
}
} Original reference: Feder, N. and O'Brien,
} T.P. (1968) Plant Microtechnique: Some
} Principles and New Methods. American Journal
} of Botany 55 (1): 123-142
}
}
}
} Dr. S. Shea Miller
} ECORC | CRECO
} Agriculture and Agri-Food Canada | Agriculture et Agroalimentaire Canada
} 960 Carling Avenue | 960, avenue Carling
} Ottawa, ON K1A 0C6
} E-mail Address / Adresse courriel shea.miller-at-agr.gc.ca
} Telephone | TÈlÈphone 613-759-1760
} Facsimile | TÈlÈcopieur 613-759-1701
} Teletypewriter | TÈlÈimprimeur 613-773-2600
Government of Canada | Gouvernement du Canada
--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Biology Department
/_ / __ /__ \ \ \__ 611 N. Pleasant St.
/ / / \ \ \ University of Massachusetts
/ / / \ \ \ Amherst, MA, 01003
/ / ___ / \ \__/ \ ____
www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243


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From: oshel1pe-at-cmich.edu
Date: Thu, 11 Nov 2010 07:37:48 -0600
Subject: [Microscopy] Fwd: Ask-A-Microscopist amylose and amylopectin

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Jon,

My hot plate is rather hot but I have not taken the temperature and it is off. The most frequent reason for my sections falling off the slide is impatience. When I think it is time to put the stain on I stop and wait another half minute. Total time would be maybe 3 minutes for I use a rather small drop of water most times. I guess at least a minute or so after the water has evaporated would be fine. I would go nuts if I waited 20 minutes.

For the staining I wait until a small ring can just be seen at the stain drop boundary, rinse immediately and dry down again. The time for this step can be shortened considerably by absorbing most of the water on the slide around the section with any type of tissue or paper towel.

OH, if a section is much thicker, say 2 microns it will, in my hands, still wash off if not treated very gently. I watch for a floating section, rescue it with a loop, pass it through several drops of water to clean off the stain and then dry it again onto the slide.

The suggestion of using treated slides is a good one.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E - Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not represent the NIH. This message is not confidential and can be freely shared and reproduced.


________________________________
X-from: {jkrupp-at-deltacollege.edu}
Reply-To: {jkrupp-at-deltacollege.edu}


} From: Agnes Höchstötter {agneshoechst-at-web.de}
} Subject: Ask-A-Microscopist
} realname - Agnes Höchstötter
} Email - agneshoechst-at-web.de
} ORGANIZATION - TU München
} EDUCATION - Undergraduate College
} LOCATION - Freising, Bavaria, Germany
} SUBJECT_OF_QUESTION - CLSM starch-staining
} QUESTION - Confocal laser scanning microscopy:
} Is it possible to visualize/stain amylose and
} amylopectin independently/separately of each
} other in a starch-sample or in cereal-based
} products?
}
--
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From: semhawk-at-yahoo.com
Date: Thu, 11 Nov 2010 12:51:28 -0600
Subject: [Microscopy] viaWWW: Gatan792/JEOL1010 serial communications problem

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Email: semhawk-at-yahoo.com
Name: Pierre A. Bustanoby

Organization: Specialized Electron Microscope Service

Title-Subject: [Filtered] Gatan792/JEOL1010 serial communications problem

Message: We are experiencing an initialization problem between
the JEOL 1010 and a Gatan 792 CCD camera.

Upon starting "Gatan Micrograph" the imaging window opens with the
following error:

"Unable to initialize microscope control:Failed to get reply for JEOL
serial command "ACCSET""

We have verified that the JEOL serial board is alive
with a turn around connector.

Has anyone run across this problem before??

Thanks!!

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From: bjacks13-at-uwyo.edu
Date: Thu, 11 Nov 2010 12:56:13 -0600
Subject: [Microscopy] viaWWW: Student Project on Inverted Microscopes

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Email: bjacks13-at-uwyo.edu
Name: Britney Jackson

Organization: University of Wyoming

Title-Subject: [Filtered] Student Project on Inverted Microscopes

Message: Hi,
My name is Britney Jackson, and I am a student at the University of
Wyoming. I am doing a project where I am trying to create a
business. It has come to my attention that there are some problems
with the oil spills on the lens objective. I am trying to create a
business concept around a product that would be placed around the
objectives to stop the oil spills. The fabric that I would use would
be an oil absorbent, and was wondering if anyone would answer a few
questions for my project.

1. How big of a problem are oil spills to you
2. How much will it cost you if your objective does get ruined?
3. What methods do you use for this problem right now, and do they
work well or would you be willing to look at another product?
4. How much time do you spend cleaning your objectives to make sure
that the oil doesn't ruin them?

If you could answer these questions for me it would help with my
project tremendously. Thank you and I hope to hear from you.

Britney Jackson

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From: naomi_mccallum-at-health.qld.gov.au
Date: Thu, 11 Nov 2010 18:05:56 -0600
Subject: [Microscopy] Re: viaWWW: Gatan792/JEOL1010 serial communications

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Hi Pierre

We had a similar problem between our JEOL JEM-1011 and Olympus Morada. Our situation was complicated by a damaged coms cable and a PC crash. So to abridge War & Peace from our experience:

1. First determine if tools have a pecking order. In our lab you have to turn on EM first, then PC (and even iTEM software before Stream software). (Yes Voodoo ritual)
2. From the JEOL, enter "ext 1" and JEOL should respond OK.
3. Then from JEOL enter "MAGCOD" and JEOL should reply OK: magcod [EM ...blah blah blah]
If you get Timeout this confirms com problem.
4. Investigate COM port and serial switch designation
Our camera software has a .ini file that designates the serial switch setting. Also, JEOL is dominant and needed to be assigned to COM1, Camera is passive and assigned to COM2. This needs to be checked in the camera software config and the PC config.

Sorry, I am a technovice so I can't use tech speak. If probs continue, you could contact JEOL and ask for advice from Head Service Engineer, JEOL Australasia.

Hope this helps,
Naomi


Naomi McCallum
Supervising Scientist, EM Unit, Anatomical Pathology
Pathology Queensland
_________________________________________________
Clinical and Statewide Services Division| Queensland Health

Block 7 Level 2
RBWH Queensland 4029
Ph: 07 3636 8057
Mob:
Fax: 07 3636 8908
Email: naomi_mccallum-at-health.qld.gov.au
Web: http://www.health.qld.gov.au/qhcss/qhps


} } } {semhawk-at-yahoo.com} 12/11/2010 5:00 am } } }



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Email: semhawk-at-yahoo.com
Name: Pierre A. Bustanoby

Organization: Specialized Electron Microscope Service

Title-Subject: [Filtered] Gatan792/JEOL1010 serial communications problem

Message: We are experiencing an initialization problem between
the JEOL 1010 and a Gatan 792 CCD camera.

Upon starting "Gatan Micrograph" the imaging window opens with the
following error:

"Unable to initialize microscope control:Failed to get reply for JEOL
serial command "ACCSET""

We have verified that the JEOL serial board is alive
with a turn around connector.

Has anyone run across this problem before??

Thanks!!

Login Host: 140.107.55.36
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From: dlowry-at-asu.edu
Date: Fri, 12 Nov 2010 00:24:34 -0600
Subject: [Microscopy] viaWWW: Leica Ultracut service

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Name: David Lowry

Organization: Arizona State University

Title-Subject: [Filtered] Leica Ultracut service

Message: we are looking for a qualified service technician to provide
a periodic maintenance for Leica Ultracut microtomes (E and R
models), location is the SW USA region.

thank you-

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From: brendan.griffin-at-uwa.edu.au
Date: Fri, 12 Nov 2010 00:25:03 -0600
Subject: [Microscopy] viaWWW: Electron microprobe position

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Email: brendan.griffin-at-uwa.edu.au
Name: Brendan Griffin

Organization: The University of Western Australia

Title-Subject: [Filtered] Electron microprobe position

Message:
The CMCA has recently installed a state-of-the-art JEOL 8530F
field-emission electron microprobe and the focus of this position is
to provide instruction and guidance to users on the operation of this
instrument and associated microanalysis software. We are seeking a
highly motivated and appropriately experienced person to be involved
with the day-to-day operation of the electron microscopy facilities
including maintenance.

For more information please go to Ref 3280 at
https://www.his.admin.uwa.edu.au/jobvacs/external/general/ads.htm

Thanks, Jeanette

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From: benada-at-biomed.cas.cz
Date: Fri, 12 Nov 2010 02:01:39 -0600
Subject: [Microscopy] Re: viaWWW: Gatan792/JEOL1010 serial communications problem

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Hello Pierre,
A long time ago we had a similar problem with serial communication between PC and Compustage of
our microscope. The trouble was in COM1 port setting. Someone/something (???) changed it and the
communication was completely lost.
Try to check "Baud rate", "Parity", "Data Bits", "Stop Bits" setting for specified COM port in
your communication protocol for PC {--} Gatan792/JEOL1010 interface talk. It should be
identical on both sides.

Best regards from Prague
Oldrich

---------------------------
Oldrich Benada
Institute of Microbiology of the ASCR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

On Thursday 11 of November 2010 19:54:20 semhawk-at-yahoo.com wrote:
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}
} Organization: Specialized Electron Microscope Service
}
} Title-Subject: [Filtered] Gatan792/JEOL1010 serial communications problem
}
} Message: We are experiencing an initialization problem between
} the JEOL 1010 and a Gatan 792 CCD camera.
}
} Upon starting "Gatan Micrograph" the imaging window opens with the
} following error:
}
} "Unable to initialize microscope control:Failed to get reply for JEOL
} serial command "ACCSET""
}
} We have verified that the JEOL serial board is alive
} with a turn around connector.
}
} Has anyone run across this problem before??
}
} Thanks!!
}
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From: eschumacher-at-mccrone.com
Date: Fri, 12 Nov 2010 08:11:35 -0600
Subject: [Microscopy] Emergency Plan: Summary of Responses

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Hello Fellow Listers,

Thanks to those of you who responded to my recent inquiry about emergency plans for restoration of equipment and services. There was quite a bit of interest in the question, and a number of good suggestions, which I've summarized below. Responses tended to fall into categories of preparation to minimize damage to equipment, and strategies for cleanup and restoration of services. Flooding seemed to be the most common scenario, though I received some input about fire damage and electrical outages as well.

On the planning and preparation side, suggestions included: generators to run sump pumps, early warning to allow for safe shutdown of equipment, sand bagging of lab doors, a phone tree for notification of lab personnel, keeping cables and control boxes off of the floor as much as possible, UPS backup of instruments coupled with a system that notifies you of power failure, allowing time to come in and shut instruments down safely while still on the UPS, and shutdown at master shutoff switches if possible, to minimize damage from a power surge at the instrument when service is restored. Consider plans for computer backup, including both data files and the customized software that you use to run instruments and analyze data. If you have insurance, what is the age of your equipment, and are you sufficiently covered for replacement value?

In terms of cleanup and restoration, consider where you might stand in the priority list for cleanup, both within your organization and with your instrument service providers. It will almost certainly be up to lab personnel to step in and deal with the brunt of the recovery operation, but bear in mind that, in the event of a large scale disaster in your area, your staff will be dealing with personal issues, too, ranging from difficulty in getting to the lab to loss of their own homes, and will need support.

Thanks to Bill Anderson for this list of things to have on hand in the aftermath of flooding, which seems to be the most common problem:

Phone numbers for physical plant or a plumber to stop the water flow
Well labeled electrical panels to know which circuit breakers to flip off
and just as important - which ones NOT to flip off in a panic situation
Plastic sheeting to throw over systems and workbenches
Wet vacs or similar vacuum cleaners that can suck up water
Electrical extension cords that won't electrocute the vacuum-er
Fans to help dry out the lab
Wedge blocks to hold doors open
Absorbent pads, sponges etc.

Bill notes that most microscopes, if installed correctly, can tolerate a few inches of water on the floor. However they should be shut off and restarted after the humidity is back to normal levels.

Though flooding is common, I got input about fire and smoke damage, difficult to deal with because it permeates everything. Remediation may involve hiring a specialized cleanup company to go over surfaces to remove soot, and use of specialized paint to seal pipes, walls and other surfaces. Delicate electronic and optical equipment will also require specialized cleanup, or may need replacement.

Finally, a general note about the instruments themselves; because we deal with equipment that is complex, expensive and has some potential associated hazards, we should operate in "what if" mode every day. Monitoring equipment performance, especially with older components, and keeping in mind what might happen during off hours (electronics failure, cooling hose leak, etc.), is a good habit to develop. One last look in the lab before you leave every day to see how the instrumentation has been left may save you some down time.

Thanks again to everyone who responded. Your interest in the question and your suggestions are very much appreciated.

All the best,

Elaine



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Elaine F.Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com



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From: vakimler-at-med.wayne.edu
Date: Sun, 14 Nov 2010 19:24:29 -0600
Subject: [Microscopy] viaWWW: Compromised Antigenicity

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Compromised Antigenicity

Message: When using LR-White Medium or Hard, does the addition of the
benzoyl peroxide compromise immunogold-conjugate antigenicity of
epitopes on thin section TEM? Some of our antibodies appear to work
and some do not.

Would it be advisable to increase the concentration (dilution) of the
one(s)that give us labeling that is weak/different from the controls?

Thanks!

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From: cogswell-at-nbnet.nb.ca
Date: Sun, 14 Nov 2010 19:24:55 -0600
Subject: [Microscopy] viaWWW: Parts for Vibrotome H1200

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Email: cogswell-at-nbnet.nb.ca
Name: Steven Cogswell

Organization: University of New Brunswick

Title-Subject: [Filtered] Parts for Vibrotome H1200

Message: Hi folks;

I've got a Vibrotome H1200 that I'm looking for parts for.
Specifically I need the stupid plastic spline that's on the bottom of
the lifting mechanism's rotator shaft. Here's a picture:

http://imgur.com/uXw4W.jpg

The company appears to have been bought and sold a dozen times since
we bought this unit. Maybe Quorum, maybe Leica own them now, but
nobody I contact at those companies knows how to get parts.

Does anyone know where to get parts? Do you have parts you'd like to sell me?

(Aside: a machine made almost entirely by milling out a huge chunk of
aluminum, and the weak plastic part has to take all the torque.
Sheesh.)

Thanks for your time,
Steven Cogswell, P.Eng.

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From: chengge.jiao-at-fei.com
Date: Sun, 14 Nov 2010 19:25:15 -0600
Subject: [Microscopy] viaWWW: Slow speed diamond saw

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Email: chengge.jiao-at-fei.com
Name: c jiao

Organization: fei

Title-Subject: [Filtered] Slow speed diamond saw

Message: Dear All

I need to find a right brand slow speed diamond saw for our sample
preparation before FIB sample for 3D EBSD. The most important thing
for me is to buy a right one. However, I am not familiar with this
kind of product.

Our sample size is usually not big, 20 mm x 20 mm, but we need to
have good cutting accuracy and finishing quality, and it is better if
the cutting thickness is not too thick as well.

It would be even nice, if you can let me know the price.

Thanks and Regards

C Jiao

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From: leunissen-at-aurion.nl
Date: Sun, 14 Nov 2010 21:15:54 -0600
Subject: [Microscopy] Re: viaWWW: Compromised Antigenicity

Contents Retrieved from Microscopy Listserver Archives
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Hello Vickie,

I can not answer whether benzoyl peroxide is the culprit, as it is hard to isolate causes in your case.

Any treatment may impair epitope recognition and different epitopes may be sensitive to different treatments. Sometimes this is because of chemical modification, other times because of steric hinderance as a result of for instance cross linking. Resins may mask epitopes by encapsulating them and these are just a few examples. These issues are not easily covered in an email but they are often covered in immuno workshops that are announced in this forum on a regular basis. You could check the archives.

Increasing primary antibody concentration to boost the signal may sometimes help. But keep in mind that you can get any protein to interact with any other protein or macromolecule as long as your concentrations are high enough. In other words: the reliability that what you detect is actually 100% antigen may decrease with increasing antibody concentration. I know sometimes high concentrations are used, but this is more helping to achieve a fast result than that it should be required to get a result in the first place. These antibodies of course need to be 'cooperative', i.e. not sticky and of high specificity.

As a rule an antibody concentration of 1-10µg/ml should be more than sufficient. In my experience concentrations between 0.1 and 1 µg/ml are preferred. If that does not reveal a consistent pattern, then it is likely that the antigen is not detectable in sections and you may need to resort to a different fixation, different preparation method.

It is usually not easy or even possible to get consistently positive detections with just one prep technique and as usual such issues need to be approached from a few different angles....

Please feel free to contact me off-list if you need more info.


Jan Leunissen
Aurion
http://www.aurion.nl

On 15/11/2010, at 2:24 PM, vakimler-at-med.wayne.edu wrote:

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} Title-Subject: [Filtered] Compromised Antigenicity
}
} Message: When using LR-White Medium or Hard, does the addition of the
} benzoyl peroxide compromise immunogold-conjugate antigenicity of
} epitopes on thin section TEM? Some of our antibodies appear to work
} and some do not.
}
} Would it be advisable to increase the concentration (dilution) of the
} one(s)that give us labeling that is weak/different from the controls?
}
} Thanks!
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From: benada-at-biomed.cas.cz
Date: Mon, 15 Nov 2010 02:55:53 -0600
Subject: [Microscopy] Re: viaWWW: mapping montage misfit with Noran System Six

Contents Retrieved from Microscopy Listserver Archives
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Hello Lindsay,
I do not know anything about the Noran System Six, but we had similar problem on our TEM Philips
CM100 and AnalySis system. We were not able to do proper image stitching (montage) via movement
of the stage. We also had some gaps between individual images in final composed image. The
problem was in the communication speed between PC and the microscope.
Our system communicates with the stage through the serial line (RS-232) and the default baud
rate setting of COM port in WinXP was 115200. When we changed it to 9600 the image stitching
started to work.

Best regards
Oldrich

---------------------------
Oldrich Benada
Institute of Microbiology of the ASCR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic


On Tuesday 09 of November 2010 22:44:04 you wrote:
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} Name: L. Keller
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}
} Title-Subject: [Filtered] mapping montage misfit with Noran System Six
}
} Message: We are using a Noran SDD on our FEG SEM to produce spectrum
} images of rock thin sections. One issue we are having is that in
} assembling montages of large areas, there are gaps between the
} individual component images - we've double checked the calibration a
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==============================Original Headers==============================
6, 23 -- From benada-at-biomed.cas.cz Mon Nov 15 02:55:53 2010
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From: mary.anton-at-srs.gov
Date: Mon, 15 Nov 2010 08:40:32 -0600
Subject: [Microscopy] viaWWW: Fullam Mark II carbon coater

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Email: mary.anton-at-srs.gov
Name: Mary Anton

Organization: SRNL

Title-Subject: [Filtered] Fullam Mark II carbon coater

Message: I have a Fullam Mark II carbon coater that is currently not
working. I do not have any schematics on it and wonder if anyone
does. If not, does someone know what fuses it requires? Pella did
not buy Fullam's coater line and this information is now not
available through the manufacturer. Thanks for your help.


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==============================Original Headers==============================
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From: larry.ackerman-at-ucsf.edu
Date: Mon, 15 Nov 2010 12:44:43 -0600
Subject: [Microscopy] Re: viaWWW: Parts for Vibrotome H1200

Contents Retrieved from Microscopy Listserver Archives
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Hi Steven,
Your instrument is apparently a Bio-Rad Laboratories Micro-Cut H1200
Vibratome probably also marketed by others such as EMS. I saw a couple
units for sale on dotmed.com and medwow.com but maybe your best bet is
to contact the Vibratome experts at:

http://www.vibratome.com

Good luck,
Larry


cogswell-at-nbnet.nb.ca wrote:
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} Email: cogswell-at-nbnet.nb.ca
} Name: Steven Cogswell
}
} Organization: University of New Brunswick
}
} Title-Subject: [Filtered] Parts for Vibrotome H1200
}
} Message: Hi folks;
}
} I've got a Vibrotome H1200 that I'm looking for parts for.
} Specifically I need the stupid plastic spline that's on the bottom of
} the lifting mechanism's rotator shaft. Here's a picture:
}
} http://imgur.com/uXw4W.jpg
}
} The company appears to have been bought and sold a dozen times since
} we bought this unit. Maybe Quorum, maybe Leica own them now, but
} nobody I contact at those companies knows how to get parts.
}
} Does anyone know where to get parts? Do you have parts you'd like to sell me?
}
} (Aside: a machine made almost entirely by milling out a huge chunk of
} aluminum, and the weak plastic part has to take all the torque.
} Sheesh.)
}
} Thanks for your time,
} Steven Cogswell, P.Eng.
}
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} ==============================Original Headers==============================
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} .
}

--
Larry Ackerman, Specialist
Electron Microscopy Lab
Manager, DERC Microscopy Core
UCSF, Dept. of Anatomy, Rm S1355
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

415-999-4758

==============================Original Headers==============================
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From: beth-at-plantbio.uga.edu
Date: Mon, 15 Nov 2010 16:06:36 -0600
Subject: [Microscopy] DNA extraction after SEM?

Contents Retrieved from Microscopy Listserver Archives
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Hi all,
Does anyone know if it is easy to do DNA extraction on insects after
they have been sputter-coated with gold or gold palladium? Or, is it
better to sacrifice a leg beforehand? Theirs, not mine;-)
thanks for any advice,
Beth

} I have one more question: are the samples still usable for DNA
} extraction afterwards or should we take a few legs off just to be
} sure?


==============================Original Headers==============================
3, 19 -- From beth-at-plantbio.uga.edu Mon Nov 15 16:06:36 2010
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From: nizets2-at-yahoo.com
Date: Tue, 16 Nov 2010 03:36:37 -0600
Subject: [Microscopy] DNA extraction after SEM?

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Hi Beth!

I am no insect specialist so here I only rely on common sense.
Since you are coating the mineralized external skeleton of the insect I don't
see how the coating in itself could have an impact on DNA.
The question is different if you consider the whole processing to prepare the
insect for SEM analysis: fixation and dehydration for example.
In any case, I also don't see why one should break a leg to extract DNA. Can't
you simply extract some liquid (hemolymph) with a fine needle and a syringe? If
not, ask your friendly neighbourspider to show you the way.

Regards,

Stephane



----- Original Message ----
X-from: "beth-at-plantbio.uga.edu" {beth-at-plantbio.uga.edu}
To: nizets2-at-yahoo.com
Sent: Mon, November 15, 2010 11:11:10 PM

Hi all,
Does anyone know if it is easy to do DNA extraction on insects after 
they have been sputter-coated with gold or gold palladium? Or, is it 
better to sacrifice a leg beforehand? Theirs, not mine;-)
thanks for any advice,
Beth

} I have one more question: are the samples still usable for DNA 
} extraction afterwards or should we take a few legs off just to be 
} sure?


==============================Original Headers==============================
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From: delannoy-at-jhmi.edu
Date: Tue, 16 Nov 2010 08:50:52 -0600
Subject: [Microscopy] neg stain in sections?

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Hello Members,
I have a user who ran some collagen sheets, standard TEM. The samples got both osmium and uranyl acetate (after primary fixation). When we double stain afterwards we see mostly a negative stained image of the sectioned collagen. We have tried both aqueous and methanolic uranyl acetate followed by lead citrate, but still keep getting white fibers (neg stained on sections). Any suggestions as to what is happening and a remedy will be greatly appreciated. We have also sectioned from 60 to 80 nm thickness.

Thanks,
M Delannoy

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From: rgore11-at-hotmail.com
Date: Tue, 16 Nov 2010 09:34:05 -0600
Subject: [Microscopy] viaWWW: TEM section sink into =?iso-8859-1?Q?liquid=9C?=

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Email: rgore11-at-hotmail.com
Name: Rick Gore

Title-Subject: [Filtered] TEM section sink into liquidœ

Message: Dear all,

I am trying to prepare TEM
section (~50nm thick) using the
wet-cryo-ultramicrotomy technique. The liquid I
use is DMSO/H2O mixture(60:40 in v). I used "wet"
diamond cryo knife. But instead of floating on
the surface of the liquid, my TEM section sink
into the liquid as soon as it was cut. Would you
please help me out?

Alternatively, I have tried "dry"
cryo-ultramicrotomy technique. But the TEM
sections curl like hell...So I prefer the
wet-cryo-ultramicrotomy technique, but any
suggestion on how to make the section flat is
highy appreciated.

Thanks a lot for your consideration.

Sincerely,
Rick Gore


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From: delannoy-at-jhmi.edu
Date: Tue, 16 Nov 2010 09:50:22 -0600
Subject: [Microscopy] naeg stain in sections

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Forgot to mention, UA stained only and unstained sections show the same negative image.

MD

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From: W.Muss-at-salk.at
Date: Tue, 16 Nov 2010 09:56:40 -0600
Subject: [Microscopy] AW: neg stain in sections?

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Dear M. Delannoy,

from older (and somehow specific) literature and some personal comments made by scientists like Feroze N. Ghadially in the late 80ies, I know that the problem originates with fixation (FA-GA, Osmium), washes in buffer and perhaps also ethanols used for dehydration of TEM - tissue preparations like liver, kidney and others.

In earlier times (when a "novice", as Ghadially states) I faced the problem of "negatively" stained collagen fibres/fibrils more often, especially in diseased tissues, but have not had such an adverse effect any more in specimens treated with paraphenylenediamine in 70% Etanol (after osmication, following the 50% EtOH dehydration step).
It also could be there were alterations in the quality of your staining solutions used so far (pH UO2Ac. & Pbcitrate as well?) and, if I remember correctly, perhaps also somehow altered resin quality or structural/biochemical alteration of the fibril composition was/were reported to be a candidate for that staining problem.

E.g., Feroze N Ghadially in his famous books
{Ultrastructural Pathology of the Cell and matrix { (4th ed, Volume One, p.144; Butterworth-Heinemann,Boston... 1997) in general comments on:
} Misinterpretation of viral particles {
"...Structures mistakenly interpreted as virus or as virus-like particles include: 1) , 2)., .., ....,
16) cross-cut poorly stained collagen fibrils where only the periphery is stained (so called "hollow collagen", "tubular collagen" or "negatively stained collagen").... (end of quote)

Unfortunately he has not given a comprehensive comment for that staining problem.... but I remember there also were some articles on that problem in the journal {Ultrastructural Pathology} .

On page 1308 (Volume Two, see above), Ghadially writes:
"...A more pronounced version of the same artifact [hollow collagen, where only the periphery of the fibril(s) is/are stained] is the socalled negatively stained collagen, where the collagen fibrils have failed to stain and appear quite lucent against a darker matrix. It is possible that, in some circumstances, negatively stained collagen and "tubular collagen" may reflect a pathologically altered collagen or an altered interfibrillary matrix that hampers the staining of the collagen, but to the best of my knowledge no proven example of this exists."==} cf. plate 564, Fig 2 (p. 1310) (end of quote )

Interesting problem.... I will go through my files to find any hint - but this would need a little bit time.

Meanwhile one COULD destain left unstained grids of the misstained sections (they must not have been exposed to the e-beam!) by washing cautiously e. g. in 0.2 N NaOH.....[washing afterwards carefully with a.dest.) and then try double-staining again,

perhaps you can "prestain"[i.e. before UO2Ac followed by Lead-citrate] additionally with 0.05-0.5 % hydrous Tannic acid (low molecular weight)-solution [ultrasonic dissolution and filtered befor use] for say 5-8 min -at-room temperature. Perhaps this will rescue the "normal, positive contrast staining" of your collagen fibrils.

It would be interesting to get some specific information on "sample", fixation, dehydration, resin as well as staining solutions you have used for the particular speciemn preparation.


Best wishes and good luck,


Wolfgang MUSS PhD
Salzburg, Austria




} -----Ursprüngliche Nachricht-----
} Von: delannoy-at-jhmi.edu [mailto:delannoy-at-jhmi.edu]
} Gesendet: Dienstag, 16. November 2010 15:55
} An: Muß Wolfgang
} Betreff: [Microscopy] neg stain in sections?
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} Hello Members,
} I have a user who ran some collagen sheets, standard TEM.
} The samples got both osmium and uranyl acetate (after primary
} fixation).
}
} When we double stain afterwards we see mostly a negative stained image
} of the sectioned collagen.
}
} We have tried both aqueous and methanolic uranyl acetate followed by
} lead citrate, but still keep getting white fibers (neg stained on
} sections).
}
} Any suggestions as to what is happening and a remedy will be greatly
} appreciated. We have also sectioned from 60 to 80 nm thickness.
}
} Thanks,
} M Delannoy
}
}
}
} ==============================Original
} Headers==============================
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From: raristau-at-ims.uconn.edu
Date: Tue, 16 Nov 2010 10:27:32 -0600
Subject: [Microscopy] JEM 2010

Contents Retrieved from Microscopy Listserver Archives
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I am looking for advice, instructions, warnings, etc. about attempting a
full alignment on our ten-year-old JEOL JEM2010 with FasTEM. Something that
goes far beyond basic alignments one does each day; information not in the
instruction manual included with the microscope.

Specifically, When should one use/not use free lens controls? How to correct
image shift when magnification is changed? Why do select area diffraction
pattern come from an area outside the region selected by the SA aperture?
These, and other issues, I presume could be resolved if a complete alignment
were done on a semi-annual basis.

If someone has a description of what turning each of the alignment knobs
actually does to each lens/coil, that also would be very helpful. I am
attempting to compile a table of these relations by observing lens current
changes while turning each knob, but this has proved very tedious and
imprecise.

Why am I needing this information? 1) We are a user facility with 20+
operators of various skill levels. 2) We don't carry service contracts on
our microscopes so I can't call for help every time the beam gets lost. My
institution is willing to pay for on-demand service when necessary, but I
consider keeping a microscope "in tune" something that should be within the
bailiwick of the on-site supervisor.

Cheers
Roger

Roger A. Ristau, PhD
Electron Microscopy Specialist
Institute of Materials Science
97 North Eagleville Road
University of Connecticut
Storrs, CT 06269
vox: 860-486-5453
fax: 860-486-4745






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From: rcsencsits-at-lbl.gov
Date: Tue, 16 Nov 2010 11:00:59 -0600
Subject: [Microscopy] Re: JEM 2010

Contents Retrieved from Microscopy Listserver Archives
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Hi Roger,

Quite right, when done correctly and and saved, the alignments on a JEOL TEM are a simple button touch (green N button under plastic cap, or computer recall) to recall and do not change much over the many months and I would say years. I am swamped right now but can try to help you off-line tomorrow. Free lens is never needed for regular use.
Deflectors are what you need to track and set in a spreadsheet for referrals. If a battery to memory fails you may lose the values so it is wise to have them written down.

Roseann Csencsits, PhD
Manager Donner TEM Facility
Lawrence Berkeley Lab 01-365
1 Cyclotron Road
Berkeley, CA 94720
510-486-4548









On Nov 16, 2010, at 8:34 AM, raristau-at-ims.uconn.edu wrote:

}
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} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
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} I am looking for advice, instructions, warnings, etc. about attempting a
} full alignment on our ten-year-old JEOL JEM2010 with FasTEM. Something that
} goes far beyond basic alignments one does each day; information not in the
} instruction manual included with the microscope.
}
} Specifically, When should one use/not use free lens controls? How to correct
} image shift when magnification is changed? Why do select area diffraction
} pattern come from an area outside the region selected by the SA aperture?
} These, and other issues, I presume could be resolved if a complete alignment
} were done on a semi-annual basis.
}
} If someone has a description of what turning each of the alignment knobs
} actually does to each lens/coil, that also would be very helpful. I am
} attempting to compile a table of these relations by observing lens current
} changes while turning each knob, but this has proved very tedious and
} imprecise.
}
} Why am I needing this information? 1) We are a user facility with 20+
} operators of various skill levels. 2) We don't carry service contracts on
} our microscopes so I can't call for help every time the beam gets lost. My
} institution is willing to pay for on-demand service when necessary, but I
} consider keeping a microscope "in tune" something that should be within the
} bailiwick of the on-site supervisor.
}
} Cheers
} Roger
}
} Roger A. Ristau, PhD
} Electron Microscopy Specialist
} Institute of Materials Science
} 97 North Eagleville Road
} University of Connecticut
} Storrs, CT 06269
} vox: 860-486-5453
} fax: 860-486-4745
}
}



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From: germpore-at-sonic.net
Date: Tue, 16 Nov 2010 11:12:47 -0600
Subject: [Microscopy] Re: DNA extraction after SEM?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

beth-at-plantbio.uga.edu wrote:

} Hi all,
} Does anyone know if it is easy to do DNA extraction on insects after
} they have been sputter-coated with gold or gold palladium? Or, is it
} better to sacrifice a leg beforehand? Theirs, not mine;-)
} thanks for any advice,
} Beth
}
} } I have one more question: are the samples still usable for DNA
} } extraction afterwards or should we take a few legs off just to be
} } sure?

I don't know about insects per se, but I do know that there has been
at least one experiment demonstrating that fully dehydrated and coated
fungal spores examined under SEM (not ESEM, I should point out) could
retain viability and later be germinated:

http://dx.doi.org/10.1016/0147-5975(91)90013-4

That would imply that certainly imply that intact DNA sequences could
be recovered from a specimen prepared for SEM. Then again, fungal
spores are particularly resistant structures and your milage may vary
with other kinds of specimens.

I would worry, of course, about the potential for damage to DNA
sequences from the process and amplification of damaged sequences
during PCR. However, presuming you've amplified an adequate number of
existing copies of the sequence to begin with, the damaged DNA
sequences should simply represent background noise. (And I might be
being presumptuous to assume you would be sequencing the DNA after
extraction.)

Peter Werner
Merritt College Microscopy Program
http://www.merritt.edu/apps/comm.asp?Q=40718

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From: hanke-at-mee-inc.com
Date: Tue, 16 Nov 2010 16:19:59 -0600
Subject: [Microscopy] Re: viaWWW: Slow speed diamond saw

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Most of the metallography and microscopy supply companies
can supply a slow-speed saw that will suit your needs. I
think that the functionality of the low-end versions of
these saws is about the same. The biggest differences are
probably fit and finish and in the sample holding vices that
are available. Some higher speed models are available from
some sources that may interest you. I suggest that you
contact your local sales reps for Buehler, Struers, and
Allied HighTech, as well as look on line at offerings from
South Bay, Ladd and others to determine what features and
cost best fit your situation.

--
Larry D. Hanke, P.E.
Materials Evaluation and Engineering, Inc.
Practical Solutions Through Technology and Innovation
http://www.mee-inc.com (763) 449-8870


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From: nicholls-at-uic.edu
Date: Tue, 16 Nov 2010 16:32:03 -0600
Subject: [Microscopy] Re: JEM 2010

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Roger

The simple answer to your questions are:-

For basic use of the microscope Free Lens Control should not be used at
all! FLC turns the microscope into an interesting optical bench but is not
needed for standard use of the microscope.

Image shift with magnification change is usually the result of condenser
alignment issues and can usually be addressed by selecting the appropriate
alignment on the right hand draw and returning the value to the last
engineer set one by pressing the green N button on the right hand side of
the draw. SAD patterns not coming from the same area as in Imaging mode is
also probably the result of the same alignment being incorrect.

To return all alignments to the last engineer setting select each in turn
and press N. However I suggest noting all values down (preferably in
hexadecimal - on FasTem server select Maintenance } System Status } System
Maintenance Page } Maintenance DA/C to put all DAC values onto Microscope
Screen (select System Main page } P1 Main to get back to original setting)).

All of the alignment coils controls the alignment coils in the column.
Shifts shift the bean and DEF tilt the beam. Shifts apply equal but
opposite current to the upper and lower coils while DEF apply different
currents to upper and lower coils.

Any deeper alignment than this really does need an engineer and has to be
done outside of FasTem.

Alan



At 10:28 AM 11/16/2010, you wrote:



} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America

Alan W Nicholls, PhD
Interim Associate Director - RRC
Director of Research Service Facility (Electron Microscopy)
Research Resources Center - East (M/C 337)
Room 110 Science and Engineering South Building
The University of Illinois at Chicago
845 West Taylor St
Chicago, IL 60607-7058

Tel: 312 996 1227
Fax: 312 996 8091
Office: Room 110

Web site www.rrc.uic.edu


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From: rschmitz-at-uwsp.edu
Date: Tue, 16 Nov 2010 17:57:38 -0600
Subject: [Microscopy] Trouble staining grids

Contents Retrieved from Microscopy Listserver Archives
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Dear members of the list server:

My students and I are having trouble staining our grids. I have been using
50% ethanolic uranyl acetate (Dawes 1988) and Venable and Coggesall lead
citrate since 1990's. Suddenly neither stain works, the grids show no sign
of staining and I am not even getting precipitate artifact. I have
rechecked my recipes for error. Checked the double distilled water from two
different sources and none of my colleagues doing molecular procedures
report problems with the water. Glassware is acid washed, and we use new
glass scintillation vials and new dispensing syringes (plastic) with
millipore filters to double filter the stain before using. Boiled the water
to remove CO2. We have tried different batches of uranyl acetate and lead
citrate. We have also varied staining times from 5 to 30 minutes, all with
no success.

Does any one have any idea of what to check next. As I have said I have
used this methods for staining grids successfully since the early nineties
both in my research and when teaching my undergrad course in TEM Method. To
have methods stop working out of the clear blue sky is baffling.

I am about to start trying different recipes for stain.

Any questions, comments and/or suggestion would be greatly appreciated

Bob
Dr. Robert J. Schmitz
Associate Professor of Anatomical Sciences
Department of Biology
University of Wisconsin-Stevens Point
800 Reserve Street, 380 TNR Bld
Stevens Point, WI 54481
715-346-2420
Email: rschmitz-at-uwsp.edu
http://www.uwsp.edu/biology/faculty/rschmitz/index.html




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From: j.knowles-at-ucl.ac.uk
Date: Tue, 16 Nov 2010 20:39:41 -0600
Subject: [Microscopy] viaWWW: TEM EDX 9800 system video outputs

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Email: j.knowles-at-ucl.ac.uk
Name: Jonathan Knowles

Organization: University College London

Title-Subject: [Filtered] TEM EDX 9800 system video outputs

Message: Hi

Does anybody have any idea about the video outputs from the EDX 9800
system. To explain, we had this detector in a box and put it on the
TEM to see if it all works and amazinlgy does. SO what we wanted to
do was try and take the video output from the EDX and read/grab it
via a Matrox card. This would allow us to get rid of both the massive
monitor and the the serial port printer.

I can see a ghostly image via the matrox software if I connect one of
the RGB cables but the timing is not right so the image is low
quality and distorted. I thought rather than empirically trying all
different settings (which with the Matrox cards is a large number!) I
would ask first.

Thanks

Jonathan

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From: weisse-at-mskcc.org
Date: Tue, 16 Nov 2010 20:40:16 -0600
Subject: [Microscopy] viaWWW: SEM Critical Point Dryer

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Email: weisse-at-mskcc.org
Name: Elizabeth Weiss

Organization: CUNY Hunter College

Title-Subject: [Filtered] SEM Critical Point Dryer

Message: Hunter College Biological Sciences is looking to procure an
SEM Critical Point Dryer for upcoming courses, preferably an EMS 850,
but others would be considered. Please respond ASAP as time is of
the essence. Liz

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From: pmachado-at-igc.gulbenkian.pt
Date: Tue, 16 Nov 2010 20:41:11 -0600
Subject: [Microscopy] viaWWW: TEM project on imaging

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Email: pmachado-at-igc.gulbenkian.pt
Name: Pedro Machado

Organization: IGC

Title-Subject: [Filtered] TEM project on imaging

Message: Dear List Members,

I am a TEM technician working with biological samples and taking a
masters on technology and digital art.
I would like to do my masters project on TEM imaging. I am still
going through the literature and trying to gather more data.

I would like to ask you what kind of software applications you would
like to have for your TEM when working on biological samples (would
you like to have a pattern recognition software for your favourite
organelle? would you like your TEM to gather all the fields in a
grids and present the data to you in a googlemaps fashion?etc?).

Any opinion/advice would be really useful as well as suggesting
software that has this or that useful function (or lacks something
you would really like to have).

Currently I am using a JEOL 100 CX II TEM without digital camera.
We are still preparing a room for a new Hitachi 7650, so I never got
to use a TEM with a digital interface enough to know its weak points
and explore them.

Thank you all,
Pedro Machado



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From: leunissen-at-aurion.nl
Date: Wed, 17 Nov 2010 02:36:52 -0600
Subject: [Microscopy] =?iso-8859-1?Q?Re=3A_=5BMicroscopy=5D_viaWWW=3A_TEM_section_sink?=

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Hello Rick,


Using the wet-sectioning technique developed by the legendary Prof. Wilhelm Bernhard and colleagues Leduc, Kuhlmann and Viron it can not be avoided that cryo-sections go under the liquid, at least partially. There is no surface tension to support the sections. Usually you can see some ripples in the liquid surface indicating where the sections are located. The sections can be picked up with Marinozzi rings and stretched in these rings to some extent, but not by much. Sections cut from gelatin embedded specimens curl quite a bit as a result from them hitting the viscous DMSO/Water trough liquid.

The Tokuyasu technique in general gives much flatter sections under proper conditions, but to elaborate on this would be much more appropriate for other list members. Bit rusty....


Jan





} -----
} Email: rgore11-at-hotmail.com
} Name: Rick Gore
}
} Title-Subject: [Filtered] TEM section sink into liquidË›
}
} Message: Dear all,
}
} I am trying to prepare TEM
} section (~50nm thick) using the
} wet-cryo-ultramicrotomy technique. The liquid I
} use is DMSO/H2O mixture(60:40 in v). I used "wet"
} diamond cryo knife. But instead of floating on
} the surface of the liquid, my TEM section sink
} into the liquid as soon as it was cut. Would you
} please help me out?
}
} Alternatively, I have tried "dry"
} cryo-ultramicrotomy technique. But the TEM
} sections curl like hell...So I prefer the
} wet-cryo-ultramicrotomy technique, but any
} suggestion on how to make the section flat is
} highy appreciated.
}
} Thanks a lot for your consideration.
}
} Sincerely,
} Rick Gore
}



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From: bradbury.arcana-at-btinternet.com
Date: Wed, 17 Nov 2010 07:01:07 -0600
Subject: [Microscopy] viaWWW: SEM and Micrometeorites

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Email: bradbury.arcana-at-btinternet.com
Name: dave bradbury

Organization: retired

Title-Subject: [Filtered] SEM and Micrometeorites

Message: Hi All. Any one out there can help an old timer with a
problem with an old SEM. my oldie SEM is an ISI Akashi mini TV SEM.
some very carless person has shredded the cables, I am looking for a
circuit diagram for the above Model MRS 22.
Also has anyone any info on SEM prep for micrometeorites, I have
developed a method of collecting good yeilds of the same, will share
the method to any interested Free.
If you can help ..Many thanks.
Dave Bradbury...UK

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From: corrie.van-hoek-at-tatasteel.com
Date: Wed, 17 Nov 2010 07:23:49 -0600
Subject: [Microscopy] viaWWW: Thin Film analysis with GMRFILM

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Email: corrie.van-hoek-at-tatasteel.com
Name: Corrie van Hoek

Organization: Tata Steel Europe

Title-Subject: [Filtered] Thin Film analysis with GMRFILM

Message: I try to determine the thickness of an Fe-oxide layer on an
iron substrate.
K-ratio's of O and Fe are obtained. Although the help message of the
software indicates that thickness of layer containing an element
present in layer and substrate can be calculated, I have not
succeeded to find the proper way to do this.
Who can give me the exact instructions which options I need to select
and what I need to enter?

Thanks for working with me on this subject.

Corrie

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From: kjl226-at-vt.edu
Date: Wed, 17 Nov 2010 07:25:42 -0600
Subject: [Microscopy] viaWWW: contact information for TEM sales representatives

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Email: kjl226-at-vt.edu
Name: Kathy J. Lowe

Organization: Vet. School at Virginia Tech

Title-Subject: [Filtered] contact information for TEM sales representatives

Message: I am trying to get in contact with FEI and Hitachi sales
representatives. I have called and emailed both companies but have
not received a response yet.
Can anyone help me with contact information of sales representation
on the east coast?

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From: bigelow-at-umich.edu
Date: Wed, 17 Nov 2010 11:29:59 -0600
Subject: [Microscopy] Backing pump

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Bob,

Have you changed the type of filters that you are using?
I had a similar problem about a year ago and found that when I did not use a filter on my lead stain syringe from a new box of filters the stain was fine (Millex - GV [yellow] PVDF 22. Micron Low protein binding Durapore membrane for sterilization of aqueous solutions).
I went back to using the old type filter and have had no problems since. I am currently using Millex-GS [blue] MCE - Millipore MF Membrane for sterilization of aqueous solutions. The above two types of syringe filters are available to me from my supply room and I mention the description for comparison reasons only. I know that there are similar filters from different companies that also work very well.

I did try several different lead stains before I thought about the filters being from a different box. I had picked up the different type of filters because the blue ones were out of stock and I thought that one 0.22 micron filter for sterilization of aqueous solutions was as good as another. HA!

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E - Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not represent the NIH. This message is not confidential and can be freely shared and reproduced.


________________________________
X-from: {rschmitz-at-uwsp.edu}
Reply-To: {rschmitz-at-uwsp.edu}

The question: Will replacing the oil-sealed rotary vane backing
pump with an oil-free scroll pump reduce contamination problems from
oil backstreaming in our electron microscope that is equipped with an
oil diffusion pump as the main vacuum pump?

The Answer: No, I don't think replacing the rotary vane pump with a
dry pump, and retaining the oil diffusion pump, would do any good.
Oil diffusion pumps are such a persistent source of oil vapors, that
there would probably still be problems. This matter is discussed in
come detail in Section 5.5 (p. 190) of my book, Vacuum Methods in
Electron Microscopy.. Here's a short quote:

"Fundamentally, however, there must always be molecules of the pump
fluid in the inlet of the pump at a concentration corresponding to
the vapour pressure of the fluid at the temperature prevailing
there. These molecules have random, undirected motion and are
therefore free to migrate into the vacuum system."

You can, of course, capture molecules moving from the pump into the
vacuum system by installing a liquid nitrogen trap between the pump
and the system; HOWEVER, any time that trap is allowed to warm up to
room temperature the molecules of pump fluid that have collected on
it will still be able to migrate into the system.

Actually, if you are willing to make a substantial modification of
your vacuum system the best approach might be to replace the oil
diffusion pump with a turbomolecular, or turbo-drag, pump, because,
as discussed in Section 6.1.4 of Vacuum Methods, these pumps are
virtually free from oil backstreaming, even when backed by a common
oil-sealed rotary vane pump, if the system is properly operated!
(see p.235). If you do this, however, there will still be the
possibility of contamination from the oil molecules that have
accumulated on the interior parts of the instrument over the years it
has been operated with the oil diffusion+rotary vane pumping system.

Finally, I've been visiting Dr. Larry Allard at the Advanced
Microscopy Laboratory at the Oak Ridge National Laboratory from time
to time. There he has one of the new aberration-corrected JEOL
microscopes that is especially designed to be oil-free - turbo pumps
backed by scroll pumps and a large liquid nitrogen trap in the main
vacuum line, etc. Believe it or not, at the level of resolution of
that instrument there still are problems with carbonaceous
contamination, and this time it appears to be mostly from the
specimens themselves.

Cheers, and Happy Thanksgiving,

--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From: eric-miller-at-northwestern.edu
Date: Wed, 17 Nov 2010 14:24:15 -0600
Subject: [Microscopy] Disposal Of SOLID Osmium Waste From An Os Coater?

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We have a Filgen OPC60A Osmium coater and we're not sure how to dispose
of or recycle the waste. All the information on Os disposal I've been
able to find has been for 2-4% Os solution, not solid waste. Does anyone
have any suggestions?

--


ERiC Jay Miller
Electron Microscopist
NUANCE Center

Northwestern University
Mail: 2036 Cook Hall
Office: 1152 Cook Hall
2220 Campus Drive
Evanston, IL 60208-3108

ph: (847) 467-0789
fax: (847) 467-657

http://www.nuance.northwestern.edu


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From: protrain-at-emcourses.com
Date: Wed, 17 Nov 2010 15:05:16 -0600
Subject: [Microscopy] JEM 2010

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Hi

The reality of TEM alignment or SEM for that matter is that there are basic
procedures that are repeated to a conclusion to enable the column to be
aligned to the level that you require. Just one caution people do spend too
much time messing with instrument alignment; "if it is not broken don't fix
it"!

1. One method requires repeating two actions with sets of balancing
coils or a mechanical alignment. The most common procedure is that of
aligning the gun and illumination shifts to bringing the double condenser
system into alignment. In this procedure the gun shift is aligned when C1
is set on a large spot size and C2 is brought to focus. Then C1 is set at a
higher current, C2 brought to focus and illumination shift used to obtain
the alignment. The procedure is repeated until a common centre is achieved.
A similar procedure may be used to align the last two lenses in the column;
let's call them projectors. Centre a feature in an image on the screen with
the final projector; P2 . Determine the point when the P1 starts to take
part in the magnification process and centre the feature with the P1
alignment. Drop the magnification and centre P2 increase the magnification
and centre P1 repeating until you have a constant centre.

2. Moving back up the column another common alignment method is to
watch the image as the magnification is increased checking to see when the
next lens up the column is being activated. You may see the flash of a
diffraction spot which going up the magnification range is in one position.
Moving down the magnification range note the second position of the
diffraction spot. Having judged these two positions move the lens
(Intermediate?) to place the centre point between these two spots on the
centre of the screen.

3. The simplest method for bringing the image centre on axis throughout
the magnification range is to take note of which lens is changing as you run
up the magnification. Image shifts occur as the emphasis moves from lens to
lens as the magnification range develops. When the selected image point
takes a shift check on the lens now contributing and move the image point to
the column axis with that lens alignment facility.

4. As for movement between selected area mode and diffraction the most
common error is not applying the rule correctly. In selected area mode the
operator should use the freed up Intermediate lens to focus the selected
area aperture edge. Once the aperture is in focus then the specimen must be
brought to focus in the normal way. This double action brings the image
focus to be in the same plane as the aperture which should result in a true
representation of this area when moving to diffraction.

5 Illumination shift when changing focus is rarely an alignment fault!
The lens field from the very strong objective current overlaps the weak
second condenser field and dominates; this results as a shift. Hitachi
tried to overcome this through the top hat pole piece design reducing the
field overlap. JEOL for many years simply had a deflection coil that
deflected the beam to compensate for an objective current change and
illumination shift. Philips (now FEI) compensated for the illumination
shift due to this overlap by an alignment of the two objective pole piece
units.

6 Most instruments change the final projector through the very lowest
part of the magnification range; usually recognised by strong image rotation
unless they compensate with a lens higher up the column. On some occasion
one lens is providing magnification whilst another lens is demagnifying, all
because it is best to use a strong lens even at low magnifications.

Just one observation, having watched many hundreds of people set up an
instrument, only spend an appropriate period of time aligning an instrument.
Working at high magnification for a long period of time put effort into the
alignment. Just a quick look see then only spend minutes on the alignment.

I often compare the "urge to align" with letting the air out of your car
tires each morning and then reinflating them ready for the day. If the
microscope was aligned yesterday it will be aligned today.

Finally, keep a note of the lens currents at steps throughout your
magnification range and a note of "normal" deflection coil settings also
helps.

Enjoy

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com




-----Original Message-----
X-from: raristau-at-ims.uconn.edu [mailto:raristau-at-ims.uconn.edu]
Sent: 16 November 2010 16:28
To: protrain-at-emcourses.com

I am looking for advice, instructions, warnings, etc. about attempting a
full alignment on our ten-year-old JEOL JEM2010 with FasTEM. Something that
goes far beyond basic alignments one does each day; information not in the
instruction manual included with the microscope.

Specifically, When should one use/not use free lens controls? How to correct
image shift when magnification is changed? Why do select area diffraction
pattern come from an area outside the region selected by the SA aperture?
These, and other issues, I presume could be resolved if a complete alignment
were done on a semi-annual basis.

If someone has a description of what turning each of the alignment knobs
actually does to each lens/coil, that also would be very helpful. I am
attempting to compile a table of these relations by observing lens current
changes while turning each knob, but this has proved very tedious and
imprecise.

Why am I needing this information? 1) We are a user facility with 20+
operators of various skill levels. 2) We don't carry service contracts on
our microscopes so I can't call for help every time the beam gets lost. My
institution is willing to pay for on-demand service when necessary, but I
consider keeping a microscope "in tune" something that should be within the
bailiwick of the on-site supervisor.

Cheers
Roger

Roger A. Ristau, PhD
Electron Microscopy Specialist
Institute of Materials Science
97 North Eagleville Road
University of Connecticut
Storrs, CT 06269
vox: 860-486-5453
fax: 860-486-4745






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From: jacqueline.ayotte-at-ticona.com
Date: Wed, 17 Nov 2010 17:33:42 -0600
Subject: [Microscopy] viaWWW: VSI or AFM Estimate

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Email: jacqueline.ayotte-at-ticona.com
Name: Jackie Ayotte

Organization: Ticona Engineering Polymers

Title-Subject: [Filtered] VSI or AFM Estimate

Message: Hello Listers,

Interested in an estimate for contracting out VSI (vertical scanning
interferometry)work on polymer based material.

Second option would be AFM - so an estimate for this analysis would
also be appreciated.

Just looking for basic information (since I have no idea whatsoever)
- a price range (per sample, day, or hour) would also suffice.


Thank you to anyone who'll take a minute to forward me a general estimate.

Best Regards,
Jackie

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From: gw265-at-cam.ac.uk
Date: Wed, 17 Nov 2010 18:24:20 -0600
Subject: [Microscopy] chasing small specimens round the AFS

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Does anyone have any advice on how to not lose tiny specimens during
freeze substitution? They're currently sitting in the planchettes (hats)
in which they were high pressure frozen, at -90 in acetone + glut + tannic
acid.

While the specimens are below zero, ok, they stay in the planchette and
you can change solutions easily enough. But we're wondering if they fall
out once things go above zero, and if so, then what do you do? Is
trapping in agar an option? Seems a bad idea to go from osmium + acetone,
to acetone, to agar + water, back to acetone.... but if that's the only
way to hang onto the specimen, we'll do it.

When doing chemical fixations at room temperature we would traditionally
fix cells in solution, pellet, resuspend in washes, and eventually trap in
agar, then dehydrate the agar.

Any advice greatly appreciated - this part of the process has always been
entrusted to experienced techs before, and I've been stupid enough to fail
to ask what they did. The techs where I'm working have only worked with
bigger bits of tissue!

thanks

Giselle

Dr Giselle Walker
University of Cambridge


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From: reinhard.rachel-at-biologie.uni-regensburg.de
Date: Thu, 18 Nov 2010 03:12:45 -0600
Subject: [Microscopy] chasing small specimens

Contents Retrieved from Microscopy Listserver Archives
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Dear Giselle,
you may consider to embed your specimen before doing HPF into a container,
like cellulose capillaries - as suggested in 1994 by H.Hohenberg, and later
modified and employed by him and also by us. This may simplify the handling of
your specimen.
Whether you can insert your specimen now, at 0°C, into the capillaries? I
don't know - I have never thought about this. Before HPF is the way to go, for
me.
see: Hohenberg et al. 1994 J Microscopy 175:34-43
Kuret et al. 1996 J Comput Assisted Microsc 8: 261-262
Tiedemann et al. 1998 J. Microscopy 189: 163-171
and
Rieger et al. 1995 J. Struct Biol 1995: 78-87
Rieger et al. 1997 Arch Microbiol 168: 373-379

kind regards,
Reinhard

--
PD Dr. Reinhard Rachel
Universitaet Regensburg
Centre for EM / Anatomy
Faculty for Biology&Preclin. Med.
Universitaetsstr. 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29

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From: baskin-at-bio.umass.edu
Date: Thu, 18 Nov 2010 06:59:27 -0600
Subject: [Microscopy] Re: chasing small specimens round the AFS

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Hi Giselle,
I don't have any experience with high-pressure freezing so I
am going to extrapolate from my experience with regular pressure
freezing. We have ameliorated (I won't say solved) problems with
falling off samples in the past by making the substrate stickier. I
suspect that if you put your planchetts in a plasma for 30 sec or a
minute before using them, they will be stickier (of course they have
to be in the open state so the inside surface gets exposed to the
plasma). This cleans the grease / hydrocarbons off and makes the
surface nice and hydrophillic. At least it does for glass. If that is
not enough, after the plasma, you could allow some polyLlysine to
coat the inside of the planchette. Also you might try putting an agar
film on the surface of the planchette (I don't know exactly the
geometry so I don't know how hard or easy that might be). But the
point is that agar can be fairly sticky for samples. You are not
trapping it in agar, but allowing the sample to rest on (and
hopefully stick to) the agar surface, rather than on the metal
surface.

I am sorry these are just "educated guesses" but these kinds
of approaches have helped with ambient pressure freezing.

As ever
Tobias Baskin


At 6:24 PM -0600 11/17/10, gw265-at-cam.ac.uk wrote:

}
}
} Does anyone have any advice on how to not lose tiny specimens during
} freeze substitution? They're currently sitting in the planchettes (hats)
} in which they were high pressure frozen, at -90 in acetone + glut + tannic
} acid.
}
} While the specimens are below zero, ok, they stay in the planchette and
} you can change solutions easily enough. But we're wondering if they fall
} out once things go above zero, and if so, then what do you do? Is
} trapping in agar an option? Seems a bad idea to go from osmium + acetone,
} to acetone, to agar + water, back to acetone.... but if that's the only
} way to hang onto the specimen, we'll do it.
}
} When doing chemical fixations at room temperature we would traditionally
} fix cells in solution, pellet, resuspend in washes, and eventually trap in
} agar, then dehydrate the agar.
}
} Any advice greatly appreciated - this part of the process has always been
} entrusted to experienced techs before, and I've been stupid enough to fail
} to ask what they did. The techs where I'm working have only worked with
} bigger bits of tissue!
}
} thanks
}
} Giselle
}
} Dr Giselle Walker
} University of Cambridge
}

--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Biology Department
/_ / __ /__ \ \ \__ 611 N. Pleasant St.
/ / / \ \ \ University of Massachusetts
/ / / \ \ \ Amherst, MA, 01003
/ / ___ / \ \__/ \ ____
www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

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From: randy-nessler-at-uiowa.edu
Date: Thu, 18 Nov 2010 19:10:23 -0600
Subject: [Microscopy] viaWWW: TEM of particulates

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Email: randy-nessler-at-uiowa.edu
Name: Randy Nessler

Organization: Univesity of Iowa

Title-Subject: [Filtered] TEM of particulates

Message: For those of you who routinely image nanoparticles in your
TEM, what sample preparation tricks do you use to prevent the
nanoparticles from becoming contaminants to the movable and fixed
apertures?

TIA,

Randy

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From: rausti-at-lsuhsc.edu
Date: Thu, 18 Nov 2010 19:10:55 -0600
Subject: [Microscopy] viaWWW: protocol to do EM on fingernails

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Email: rausti-at-lsuhsc.edu
Name: Ron Austin

Organization: Dept of Pathology, LSU Medical School, Shreveport, LA

Title-Subject: [Filtered] Protocol

Message: Does anyone have a protocol to do EM on fingernails? We are
looking for melanin.

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From: I.J.Portman-at-warwick.ac.uk
Date: Fri, 19 Nov 2010 03:23:24 -0600
Subject: [Microscopy] viaWWW: TEM of particulates

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We do a lot of nanoparticle work, mostly with polymers but also with
metal and ceramics.

I've not found contamination of the apertures to be much of a problem,
the vast majority of out work is done in cryo conditions so the targets
are neatly trapped in water ice. For room temperature work all our stuff
is applied as an aqueous suspension then blotted or allowed to dry, as
long as noone puts in grids so heavily loaded that bits are flaking off
we don't get problems.

I've once had a problem with silica spheres that charged up and
scattered under the beam, a few did make it to the apertures but simply
cycling the aperture in and out a couple of times was sufficient to
dislodge them.
Cheers
Ian

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Email: randy-nessler-at-uiowa.edu
Name: Randy Nessler

Organization: Univesity of Iowa

Title-Subject: [Filtered] TEM of particulates

Message: For those of you who routinely image nanoparticles in your
TEM, what sample preparation tricks do you use to prevent the
nanoparticles from becoming contaminants to the movable and fixed
apertures?

TIA,

Randy

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From: nicanornick-at-yahoo.com
Date: Fri, 19 Nov 2010 07:57:40 -0600
Subject: [Microscopy] viaWWW: SEM - absorb. current variation

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Email: nicanornick-at-yahoo.com
Name: Cimpoesu Nicanor

Organization: Technical University "Gh. Asachi" Iasi, Romania

Title-Subject: [Filtered] SEM - absorb. current variation

Message: Hello

I have a problem with my SEM equipment marked by a absorb. current
variation after filament exchange (good centered), with a value only
of 20 pA at PC10 and big variations (from 20 to 600 for example) at
scan rate 6 or bigger or magnification bigger than 1000x. So these
variations modify my images by changing the contrast and take place
in the same time with the absorb. current variation. I clean most of
the metallic parts (anode, cathode or apertures or supports) an have
no improve, under 5 scanning rate the image doesn t variate but the
image is not very good. I think that can be an electrical problem but
for reduce amplifications or less than 6 scann rate the images
obtained are fine.

Please give me some clues about what should I re-check or what can be
the problem.

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From: kenconverse-at-qualityimages.biz
Date: Fri, 19 Nov 2010 08:41:32 -0600
Subject: [Microscopy] viaWWW: SEM - absorb. current variation

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Cimpoesu,
We probably need some more information. Are you having problems with faster
or slower scan rates? Have you checked the absorbed current behavior in a
Faraday cup with different magnifications and scan speeds? What is your
sample? What kind of SEM are you using and under what conditions?

I'm leaning towards sample charging if the problem is worse with higher
magnifications and slower scan speeds. There won't be any problem if you
check the absorbed current with a Faraday cup.

Some samples, such as quartz grains, will charge even though they are coated
because the quartz is an excellent insulator. The coating will carry off
any surface charge, but a 20 or 30 kV beam will inject a substantial charge
several microns into the grain, where it will remain for long periods of
time, unless the sample is brought to atmosphere. There are many other
types of samples that can have similar issues. Try changing your
accelerating voltage and condenser lens setting (spot size) and see if
things change.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


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Email: nicanornick-at-yahoo.com
Name: Cimpoesu Nicanor

Organization: Technical University "Gh. Asachi" Iasi, Romania

Title-Subject: [Filtered] SEM - absorb. current variation

Message: Hello

I have a problem with my SEM equipment marked by a absorb. current
variation after filament exchange (good centered), with a value only
of 20 pA at PC10 and big variations (from 20 to 600 for example) at
scan rate 6 or bigger or magnification bigger than 1000x. So these
variations modify my images by changing the contrast and take place
in the same time with the absorb. current variation. I clean most of
the metallic parts (anode, cathode or apertures or supports) an have
no improve, under 5 scanning rate the image doesn t variate but the
image is not very good. I think that can be an electrical problem but
for reduce amplifications or less than 6 scann rate the images
obtained are fine.

Please give me some clues about what should I re-check or what can be
the problem.

Login Host: 81.180.223.197
---------------------------------------------------------------------------

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From: liaonu-at-gmail.com
Date: Fri, 19 Nov 2010 11:09:58 -0600
Subject: [Microscopy] How to measure electron dose in a TEM.

Contents Retrieved from Microscopy Listserver Archives
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Dear all,

I look at some biological materials in a JEOL 2100F with a Gatan
Tridiem GIF camera. The problem I have is to figure out the electron
dose. The viewing screen gives the number of the current, but I am not
sure how accurate this number is. I was wondering if there is a
faraday cup in the GIF system with that we can measure the current.
Does anyone have an idea on this, or a manu of the GIF system?

Thanks and I would appreciate your inputs.
-
Yifeng Liao
Northwestern University
Evanston, IL 60208

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From: William.F.Tivol-at-aero.org
Date: 11/18/2010 05:27 PM
Subject: [Microscopy] viaWWW: TEM of particulates

Contents Retrieved from Microscopy Listserver Archives
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Dear Randy,
I've only looked at nanoparticles in cryo, and we assumed that too
few of them would leave the grid to be a contamination problem. I am
pretty sure that this is a good assumption for the areas of the grid that
had minimal or no exposure to the beam, but it may not be so good for the
areas that were imaged. On the other hand, the scope was not located in a
clean room, so any particles in the ambient environment could make their
way into the airlock and then into the scope.
Yours,
Bill



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Email: randy-nessler-at-uiowa.edu
Name: Randy Nessler

Organization: Univesity of Iowa

Title-Subject: [Filtered] TEM of particulates

Message: For those of you who routinely image nanoparticles in your
TEM, what sample preparation tricks do you use to prevent the
nanoparticles from becoming contaminants to the movable and fixed
apertures?

TIA,

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From: kenconverse-at-qualityimages.biz
Date: Fri, 19 Nov 2010 11:56:31 -0600
Subject: [Microscopy] viaWWW: SEM - absorb. current variation

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Dear listers,
I'm not familiar with Tescan instruments and I seem to be having trouble
figuring out how an increase in the scan rate might make the absorbed
current unstable. Please see below.

If anyone has any ideas, I'm sure Nicanor Cimpoesu would love to hear from
you.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


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Email: nicanornick-at-yahoo.com
Name: Cimpoesu Nicanor

Organization: Technical University "Gh. Asachi" Iasi, Romania

Title-Subject: [Filtered] SEM - absorb. current variation

Message: Hello

I have a problem with my SEM equipment marked by a absorb. current
variation after filament exchange (good centered), with a value only
of 20 pA at PC10 and big variations (from 20 to 600 for example) at
scan rate 6 or bigger or magnification bigger than 1000x. So these
variations modify my images by changing the contrast and take place
in the same time with the absorb. current variation. I clean most of
the metallic parts (anode, cathode or apertures or supports) an have
no improve, under 5 scanning rate the image doesn t variate but the
image is not very good. I think that can be an electrical problem but
for reduce amplifications or less than 6 scann rate the images
obtained are fine.

Please give me some clues about what should I re-check or what can be
the problem.

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From: William.F.Tivol-at-aero.org
Date: 11/19/2010 09:23 AM
Subject: [Microscopy] How to measure electron dose in a TEM.

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Dear Yifeng,
When we had this problem on our scope, the service engineer
calibrated the screen current using a Faraday cup. After that, we could
rely on the screen current measurement to calibrate the CCDs, which was
done each month. Then we could calculate the dose for the magnification
we wanted in terms of the counts/pixel on the CCD. There is a way to have
the beam hit the drift tube in the GIF, but that might not be an accurate
absolute measurement, although it can be calibrated with the Faraday cup
at the same time as the screen.
Yours,
Bill



X-from: liaonu-at-gmail.com
To: William.F.Tivol-at-aero.org






Dear all,

I look at some biological materials in a JEOL 2100F with a Gatan
Tridiem GIF camera. The problem I have is to figure out the electron
dose. The viewing screen gives the number of the current, but I am not
sure how accurate this number is. I was wondering if there is a
faraday cup in the GIF system with that we can measure the current.
Does anyone have an idea on this, or a manu of the GIF system?

Thanks and I would appreciate your inputs.
-
Yifeng Liao
Northwestern University
Evanston, IL 60208


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From: wesaia-at-iastate.edu
Date: Fri, 19 Nov 2010 12:43:21 -0600
Subject: [Microscopy] viaWWW: SEM - absorb. current variation

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You mentioned using a metallic specimen. That could be about as good as a faraday cup, assuming it is not oxidized. I would also consider a piece of graphite. We have plenty of that from rods used for carbon evaporation. We have also used graphite to make specimen stubs. Drilling a hole partway through the graphite and centering that under the beam would be a first attempt at a Faraday cup. You could also fasten an aperture over the top of the hole with some conductive paint. That would make it a very effective cup.

I am confused as well why this would be a problem at fast scan rates. Charging should be less of a problem. Are you seeing bands of changing brightness as the beam scans down the frame? That would indicate some instability. Maybe it is fast enough that it only shows as random noise at slow scans. Maybe you can post a picture out there. (I would be willing to receive one directly, but they cannot go through the list.)

Are all other conditions the same? Be sure that you have not accidentally gone to a much higher or lower beam current using a different aperture.

Could the filament be unstable? Do you have another spare filament that you could try? Maybe the first one is wandering during these early hours of use. Maybe it is a rare bad filament. That would be unlucky, but possible.

Warren

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Email: nicanornick-at-yahoo.com
Name: Cimpoesu Nicanor

Organization: Technical University "Gh. Asachi" Iasi, Romania

Title-Subject: [Filtered] SEM - absorb. current variation

Message: Hello

I have a problem with my SEM equipment marked by a absorb. current
variation after filament exchange (good centered), with a value only
of 20 pA at PC10 and big variations (from 20 to 600 for example) at
scan rate 6 or bigger or magnification bigger than 1000x. So these
variations modify my images by changing the contrast and take place
in the same time with the absorb. current variation. I clean most of
the metallic parts (anode, cathode or apertures or supports) an have
no improve, under 5 scanning rate the image doesn t variate but the
image is not very good. I think that can be an electrical problem but
for reduce amplifications or less than 6 scann rate the images
obtained are fine.

Please give me some clues about what should I re-check or what can be
the problem.

Login Host: 81.180.223.197
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From: jsb43-at-cam.ac.uk
Date: Fri, 19 Nov 2010 12:55:35 -0600
Subject: [Microscopy] Re: How to measure electron dose in a TEM.

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Yifeng,

We regularly use the GIF drift tube as a Faraday cup to measure the probe
current.

The Drift Tube connector is one of the BNC-type connectors near the neck of
the GIF. Ours is the middle one of the three for the GIF 200 and GIF2000
(it is worth checking the schematics of the GIF connections). Switch off
the Gatan Instrumentation Bin (GIB) and connect coaxial BNC lead to a
pico-ammeter. You'll nead to focus the beam down as small as possible and
use the beam shifts (imaging mode) or diffraction shifts (diffraction mode)
to steer the beam into the spectrometer.

If you need to, I can provide some photos of the arrangement next week when
I get hold of the pico-ammeter.

Yours, Dr Jon Barnard

Dept of Materials Science & Metallurgy
University of Cambridge, UK

P.S. If you use a swivel chair with your microscope, you can see (&
measure) the induced current in the GIF by moving around on it! A good way
to see if the chairs you use are incompatible with operating GIF,
especially for sensitive spectrum measurements!


==============================Original Headers==============================
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From: ThiruvengadathanR-at-missouri.edu
Date: Fri, 19 Nov 2010 17:52:16 -0600
Subject: [Microscopy] viaWWW: Request for your comments on LVEM5

Contents Retrieved from Microscopy Listserver Archives
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Email: ThiruvengadathanR-at-missouri.edu
Name: Rajagopalan Thiruvengadathan

Organization: University of Missouri Columbia

Title-Subject: [Filtered] Request for your comments on LVEM5

Message: I am a research faculty in the area of Materials Science.

I request and really appreciate your comments on LVEM5- BENCHTOP
ELECTRON MICROSCOPE from Delong America Inc.

1. How is the service after warranty?.

2. Any major/minor problems with the working of this instrument?

3. Can we get cross-sectional imaging OF THIN SPECIMENS? What resolution?

4. Is there need for sample preparation accessories?

5. How is the contrast with low density materials such as polymers
and/or porous materials?

6. The company claims that they are the only ones in the market to
provide such a BENCH-SCALE TEM microscope with optional modes for BSE
and STEM. Any comment?

7. Comment on the cost of the tool as opposed to the features in imaging modes?

8. Ultimate working resolution for inorganic specimens?

9. How easy is to align the microscope?

10. What about Biological specimens imaging using this tool?

11. Any special sample preparation procedures necessary?

12. How about thin film specimens?

I once again thank you for your time and valuable expertise/advice on
this instrument.

Thanks.

Raj.

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From: tomas.hrncir-at-tescan.cz
Date: Mon, 22 Nov 2010 03:18:27 -0600
Subject: [Microscopy] viaWWW: SEM - absorb. current variation

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Nicanor Cimpoesu,
it is strange, that you have seen instabilities at high scanning rate
only. This seems to be some high frequency noise (which gets averaged
at low scanning rate). Please send me (directly, not to the list) some
images and microscope log files as well. Perhaps we can solve it
together and then inform all list members about the result.

Have you performed automated heating and centering of the new filament
after the replacement? What is actually your heating current and
emission current?

In my opinion, there might be several other reasons for filament
instability:

1. Filament centering screws (which hold the filament inside the wehnelt
cylinder) are loosened.

2. Filament is damaged (e.g. broken wire) and it's emission current is
low - try another filament.

3. Wehnelt cylinder is dirty - the first filament lifetime was 500
hours - probably it is necessary to clean wehnelt cylinder around the
central hole.

The problem of the sample should be definitely solved by using the
metal sample (e.g. an empty sample holder) or Faraday cup - by the way,
there should be two of them on your stage (see "Stage control" window).

By the way, in a case of such problems next time do not be afraid to
contact Tescan support (support-at-tescan.cz) and attach package of
microscope log files.
Best regards,
Tomas

--
Tomas Hrncir, Ph.D.
R&D - Physics
Tescan
Libusina trida 21
623 00 Brno - CZ
Phone: +420 547 130 468
Fax: +420 547 130 415
http://www.tescan.cz

On Fri, 19 Nov 2010 12:06:37 -0600
kenconverse-at-qualityimages.biz wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver On-Line Help
} http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear listers,
} I'm not familiar with Tescan instruments and I seem to be having
} trouble figuring out how an increase in the scan rate might make the
} absorbed current unstable. Please see below.
}
} If anyone has any ideas, I'm sure Nicanor Cimpoesu would love to hear
} from you.
}
} Ken Converse
} owner
}
} QUALITY IMAGES
} Servicing Scanning Electron Microscopes
} Since 1981
} 474 So. Bridgton Rd.
} Bridgton, ME 04009
} 207-647-4348
} Fax 207-647-2688
} kenconverse-at-qualityimages.biz
} qualityimages.biz
}
}
} -----Original Message-----
} X-from: nicanor nick [mailto:nicanornick-at-yahoo.com]
} Sent: Friday, November 19, 2010 10:00 AM
} To: Ken Converse
} Subject: Re: [Microscopy] viaWWW: SEM - absorb. current variation
}
} Dear sir,
}
} I dont have now a Faraday Cup but I will get one monday morning and
} try to analyze the problem, I don t analyze a specific material , I
} use the normal stub
} provided with microscope, the model is Vega Tescan LMH II, and I am at
} 500 hours
} usage of the microscope and 3 hours of new filament (by from a Tescan
} delivery
} brand from Romania - so should be trusted). I have my absorb. current
} variations
} at high scanning rates (more than 4) when I try to obtain a proper
} image of a
} metallic material. the problem persist at 5, 10 and 20 kV filament
} power supply
} and the manual centering of the gun doesn t give s any results.
}
}
} Thank you for your kind response
}
} Assist. PhD. Eng. Nicanor Cimpoesu
} Faculty of Materials Science and Engineering
} "Gh. Asachi" Technical University from Iasi
} Bd. D. Mangeron 61A 700050 IASI
} Romania
} http://www.tuiasi.ro/facultati/sim/index.php?page=917
}
}
}
} ----- Original Message ----
} X-from: Ken Converse {kenconverse-at-qualityimages.biz}
} To: nicanornick-at-yahoo.com; MSA Listserver {microscopy-at-microscopy.com}
} Sent: Fri, November 19, 2010 4:41:18 PM
} Subject: RE: [Microscopy] viaWWW: SEM - absorb. current variation
}
} Cimpoesu,
} We probably need some more information. Are you having problems with
} faster or slower scan rates? Have you checked the absorbed current
} behavior in a Faraday cup with different magnifications and scan
} speeds? What is your sample? What kind of SEM are you using and
} under what conditions?
}
} I'm leaning towards sample charging if the problem is worse with higher
} magnifications and slower scan speeds. There won't be any problem if
} you check the absorbed current with a Faraday cup.
}
} Some samples, such as quartz grains, will charge even though they are
} coated because the quartz is an excellent insulator. The coating will
} carry off any surface charge, but a 20 or 30 kV beam will inject a
} substantial charge several microns into the grain, where it will
} remain for long periods of time, unless the sample is brought to
} atmosphere. There are many other types of samples that can have
} similar issues. Try changing your accelerating voltage and condenser
} lens setting (spot size) and see if things change.
}
} Ken Converse
} owner
}
} QUALITY IMAGES
} Servicing Scanning Electron Microscopes
} Since 1981
} 474 So. Bridgton Rd.
} Bridgton, ME 04009
} 207-647-4348
} Fax 207-647-2688
} kenconverse-at-qualityimages.biz
} qualityimages.biz
}
}
} -----Original Message-----
} X-from: nicanornick-at-yahoo.com [mailto:nicanornick-at-yahoo.com]
} Sent: Friday, November 19, 2010 9:01 AM
} To: kenconverse-at-qualityimages.biz
} Subject: [Microscopy] viaWWW: SEM - absorb. current variation
}
}
}
}
} ----------------------------------------------------------------------------
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} America To Subscribe/Unsubscribe --
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} http://www.microscopy.com/MicroscopyListserver/FAQ.html
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} This Question/Comment was submitted to the Microscopy Listserver
} using the WWW based Form at
} http://microscopy.com/MicroscopyListserver/MLFormMail.html
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} replying please copy both nicanornick-at-yahoo.com as well as the
} MIcroscopy Listserver
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}
} Email: nicanornick-at-yahoo.com
} Name: Cimpoesu Nicanor
}
} Organization: Technical University "Gh. Asachi" Iasi, Romania
}
} Title-Subject: [Filtered] SEM - absorb. current variation
}
} Message: Hello
}
} I have a problem with my SEM equipment marked by a absorb. current
} variation after filament exchange (good centered), with a value only
} of 20 pA at PC10 and big variations (from 20 to 600 for example) at
} scan rate 6 or bigger or magnification bigger than 1000x. So these
} variations modify my images by changing the contrast and take place
} in the same time with the absorb. current variation. I clean most of
} the metallic parts (anode, cathode or apertures or supports) an have
} no improve, under 5 scanning rate the image doesn t variate but the
} image is not very good. I think that can be an electrical problem but
} for reduce amplifications or less than 6 scann rate the images
} obtained are fine.
}
} Please give me some clues about what should I re-check or what can be
} the problem.
}
} Login Host: 81.180.223.197
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} ==============================Original
} Headers============================== 8, 22 -- From
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From: jmircheski-at-us.es
Date: Mon, 22 Nov 2010 05:18:45 -0600
Subject: [Microscopy] TEM distinguishing between mast cells and other granulocytes

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear List Users,

I have been working recently with mouse cells harvested by peritoneal
lavage. My object of interest are the peritoneal mast cells. In theory, they
are quite distinguishable – big, round or oval shaped cells with a nice
nucleus, big and a lot electron dense vesicles. This is fine, when the
images are of text-book quality. But, the peritoneal cells are heterogeneous
and other granulocytes are present. How to distinguish mast cells (at TEM
level) from other granulocytes especially when the images are not of the
best quality (e.g. the nucleus is not fully visible, there are less vesicles
in the section, the shape of the cell is different)? Eosinophils are easy to
identify and eliminate, but neutrophils and basophils???

I have been searching through the net to find some features that can help me
distinguish between the mast cells and other granulocytes, but with
questionable success so far.
Therefore, I’d like to ask the list if anyone had some experience working
with these cells.

Any advice is welcomed. Thanks a lot.

Best regards,

Josif

Dr. Josif Mircheski
____________________________________________________________________________
___
Instituto de Biomedicina de Sevilla (IBIS)
Hosp.Univ. Virgen del Rocío/CSIC/Univ. de Sevilla y
Dpto. Fisiologia Médica y Biofísica
Universidad de Sevilla
Facultad de Medicina
Avda. Sánchez-Pizjuán 4
41009-Sevilla
 
Phone:+34-954556103
Fax:+34-954551769
e-mail: jmircheski-at-us.es




==============================Original Headers==============================
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From: oshel1pe-at-cmich.edu
Date: Mon, 22 Nov 2010 08:55:02 -0600
Subject: [Microscopy] Fwd: Ask-A-Microscopist tomography reading list?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

} Date: Sun, 21 Nov 2010 04:40:29 -0800 (PST)
} From: Dr Chaitali Dekiwadia {dcd-at-unimelb.edu.au}
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Sunday, November 21,
} 2010 at 04:40:20 AM.
}
} realname - Dr Chaitali Dekiwadia
} Email - dcd-at-unimelb.edu.au
} ORGANIZATION - University of Melbourne
} EDUCATION - Graduate College
} LOCATION - Melbourne,Australia
} SUBJECT_OF_QUESTION - electron tomography
} QUESTION - Hi,
}
} Currently I am undertaking a training session on Electron
} tomography. I find it really difficult to understand the technique
} and reconstruction of tomogram using IMOD software. Is there any
} recommended reading material or method that would make understanding
} to electron tomography easier?
}
} Thanks
}
--
***************************************************************************************
Forwarded from "Ask a Microscopist"
Please remember that the person asking the question is likely
not a member the listserver, and
**any reply should go directly to the poster**
as well as to the list.
Using the "reply" function in your email does *not* send your answer
to the person asking the question.
Please copy their email address from their question.
****************************************************************************************

==============================Original Headers==============================
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From: underwoo-at-u.washington.edu
Date: Mon, 22 Nov 2010 13:08:21 -0600
Subject: [Microscopy] Re: TEM distinguishing between mast cells and other

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi Josif,

I agree that there can be many times in which the particular section can't give enough clues to make a definitive identification. However, the granule size between mast cells/basophils and neutrophils is as large as it gets (light level vs. EM level visible) and as you said you can easily exclude eosinophils by the crystalline forms in the granules. So that takes care of neutrophils and eosinohils but mast cells compared to basophils, that is a tough one. I thought the only difference between the two was tissue location and I know very little about any morphological markers that could distinguish between the two. Hopefully someone else may know.

Robert Underwood
University of Washington
Dermatology

On Mon, 22 Nov 2010, jmircheski-at-us.es wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear List Users,
}
} I have been working recently with mouse cells harvested by peritoneal
} lavage. My object of interest are the peritoneal mast cells. In theory, they
} are quite distinguishable ? big, round or oval shaped cells with a nice
} nucleus, big and a lot electron dense vesicles. This is fine, when the
} images are of text-book quality. But, the peritoneal cells are heterogeneous
} and other granulocytes are present. How to distinguish mast cells (at TEM
} level) from other granulocytes especially when the images are not of the
} best quality (e.g. the nucleus is not fully visible, there are less vesicles
} in the section, the shape of the cell is different)? Eosinophils are easy to
} identify and eliminate, but neutrophils and basophils???
}
} I have been searching through the net to find some features that can help me
} distinguish between the mast cells and other granulocytes, but with
} questionable success so far.
} Therefore, I?d like to ask the list if anyone had some experience working
} with these cells.
}
} Any advice is welcomed. Thanks a lot.
}
} Best regards,
}
} Josif
}
} Dr.?Josif?Mircheski
} ____________________________________________________________________________
} ___
} Instituto de Biomedicina de Sevilla (IBIS)
} Hosp.Univ. Virgen del Roc?o/CSIC/Univ. de Sevilla y
} Dpto. Fisiologia M?dica y Biof?sica
} Universidad de Sevilla
} Facultad de Medicina
} Avda. S?nchez-Pizju?n 4
} 41009-Sevilla
} ?
} Phone:+34-954556103
} Fax:+34-954551769
} e-mail:?jmircheski-at-us.es
}
}
}
}
} ==============================Original Headers==============================
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} 10, 38 -- To: {Microscopy-at-microscopy.com}
} 10, 38 -- Subject: TEM distinguishing between mast cells and other granulocytes
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==============================Original Headers==============================
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From: ibarke2-at-uwo.ca
Date: Mon, 22 Nov 2010 14:39:59 -0600
Subject: [Microscopy] FEG-SEM High Vacuum, High Current, WDS Plant Analysis

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html


Hello,
I am looking for some suggestions of how to run some FEG-SEM/WDS analyses on plant material.
I am a geologist by trade, and this will be the first time I've run plant material on the SEM so I'm seeking some helpful tips from some more experienced analysts.
We have a Hitachi SU6600 FEG-SEM with a WDS detector.  We are trying to determine the distribution of Cd and Zn in plant root structures, even though the Cd and Zn are in very low concentrations.
We would like to map the Cd and Zn using WDS. However, since they are at such low concentrations, I believe I would need very high probe current and long mapping times, which would easily destroy the plant material.  I was thinking of running the analysis at 20kV, 150nA, and under high vacuum (the WDS detector does not work in VP mode).
How have people over come sample damage for this type of analysis? Or have you done this type of analysis in a different way? How do you prep the samples (inject with conductive epoxy?), what do you coat with, and what would be the best operating conditions?
Please respond to ibarke2-at-uwo.ca
Many thanks!



Ivan R. Barker
UWO Earth Sciences MSc
ZAPLab Geochronology Research Technician
T: 519-661-2111 ext. 88397
E: ibarke2-at-uwo.ca
 





==============================Original Headers==============================
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From: matthew.weyland-at-mcem.monash.edu.au
Date: Mon, 22 Nov 2010 16:38:34 -0600
Subject: [Microscopy] Re: Fwd: Ask-A-Microscopist tomography reading list?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Chaitali,

I'd recommend the standard text for electron tomography, the book by Frank;

http://www.amazon.com/Electron-Tomography-Three-Dimensional-Visualization-Structures/dp/038731234X

This was updated fairly recently (2006) and is a good starting point. It covers
much of the subject in board strokes and has good references for the details!
IMOD has plenty of documentation;

http://bio3d.colorado.edu/imod/#Guides

But this does assume so basic knowledge (which you can get from Frank). If you
have specific technical questions relating to tomography you should be able to
get answers from the 3DEM mailing list (possibly more tomography oriented than
the Microscopy Listserver!);

https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem

Good luck,

Matthew

} realname - Dr Chaitali Dekiwadia
} } Email - dcd-at-unimelb.edu.au
} } ORGANIZATION - University of Melbourne
} } EDUCATION - Graduate College
} } LOCATION - Melbourne,Australia
} } SUBJECT_OF_QUESTION - electron tomography
} QUESTION - Hi,
} }
} } Currently I am undertaking a training session on Electron
} } tomography. I find it really difficult to understand the technique
} } and reconstruction of tomogram using IMOD software. Is there any
} } recommended reading material or method that would make understanding
} } to electron tomography easier?
} }
} } Thanks
} }

--
Dr M.Weyland, Senior Research Fellow & Electron Microscopist
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From: naomi_mccallum-at-health.qld.gov.au
Date: Mon, 22 Nov 2010 17:09:49 -0600
Subject: [Microscopy] Re: TEM distinguishing between mast cells and other

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Gentlemen and fellow listers

I have just consulted Norman F Cheville (Ultrastructural Pathology, 2nd Ed 2009) (for my own benefit also) and here are a few excerpts:
- Basophils are distinguished from connective tissue mast cells by their smaller, multilobed nuclei that have denser peripheral chromatin, less numerous and larger granules, and less complex cell surfaces that lack granule-containing dendritic processes.
- Basophil granules resemble those of mast cells
- Charcot-Leyden crystals, typical of eosinophils, have been reported in basophils. (We didn't want to hear that!)

- Mast cell ... The Golgi complex is large but endoplasmic reticulum small.
- Intermediate filaments are common and one or more lipid globules may be present, especially in activated mast cells.
- Species differences in basophill and mast cell numbers and in granule size are important to differences in inflammatory response. Granules are numerous in ...hamster but ... few in rabbit... guinea pig.
- Mast cell granules are round, membrane-bound, and heterogeneous. ... mature... homogeneous, amorphous, and dense. Granules with scrolls or concentrically arranged cords are less mature.
- some species ... compound granules have crystalloids of protein
- when ruptured... reveals small beaded filaments interpreted as heparin-protein complexes
- Mammalian mast cells and basophilshave granules that contain granzyme B (so some granules will be the same!)

Perhaps too much info can be confusing, but here it is. In diagnostic practice we look for fibroblasts in skin tissue specimens which we have to distinguish from mast cells and don't encounter as many basophils in this context. So our experience is a little different. However, the shape of the nucleus and the tell-tale immature "bunch of roses" granules are the main features used to distinguish them. But someone with more experience than I may have more sagely advice.

Regards
Naomi
} } } {underwoo-at-u.washington.edu} 23/11/2010 5:17 am } } }



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Hi Josif,

I agree that there can be many times in which the particular section can't give enough clues to make a definitive identification. However, the granule size between mast cells/basophils and neutrophils is as large as it gets (light level vs. EM level visible) and as you said you can easily exclude eosinophils by the crystalline forms in the granules. So that takes care of neutrophils and eosinohils but mast cells compared to basophils, that is a tough one. I thought the only difference between the two was tissue location and I know very little about any morphological markers that could distinguish between the two. Hopefully someone else may know.

Robert Underwood
University of Washington
Dermatology

On Mon, 22 Nov 2010, jmircheski-at-us.es wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear List Users,
}
} I have been working recently with mouse cells harvested by peritoneal
} lavage. My object of interest are the peritoneal mast cells. In theory, they
} are quite distinguishable ? big, round or oval shaped cells with a nice
} nucleus, big and a lot electron dense vesicles. This is fine, when the
} images are of text-book quality. But, the peritoneal cells are heterogeneous
} and other granulocytes are present. How to distinguish mast cells (at TEM
} level) from other granulocytes especially when the images are not of the
} best quality (e.g. the nucleus is not fully visible, there are less vesicles
} in the section, the shape of the cell is different)? Eosinophils are easy to
} identify and eliminate, but neutrophils and basophils???
}
} I have been searching through the net to find some features that can help me
} distinguish between the mast cells and other granulocytes, but with
} questionable success so far.
} Therefore, I?d like to ask the list if anyone had some experience working
} with these cells.
}
} Any advice is welcomed. Thanks a lot.
}
} Best regards,
}
} Josif
}
} Dr.?Josif?Mircheski
} ____________________________________________________________________________
} ___
} Instituto de Biomedicina de Sevilla (IBIS)
} Hosp.Univ. Virgen del Roc?o/CSIC/Univ. de Sevilla y
} Dpto. Fisiologia M?dica y Biof?sica
} Universidad de Sevilla
} Facultad de Medicina
} Avda. S?nchez-Pizju?n 4
} 41009-Sevilla
} ?
} Phone:+34-954556103
} Fax:+34-954551769
} e-mail:?jmircheski-at-us.es
}
}
}
}
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From: bnross-at-interchange.ubc.ca
Date: Tue, 23 Nov 2010 12:42:52 -0600
Subject: [Microscopy] Re: chasing small specimens round the AFS

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi Giselle,

When we have to do high pressure freezing with cells in suspension, we usually mix them in a 20% solution of BSA just before freezing. The BSA reacts with Osmium in the freeze-substitution process and most of the time you end up with a nice little solid brown/black "hockey puck" that is chock full of cells and very easy to embed and section. When you mix the cells with the BSA, try to have the cells as concentrated as possible in the BSA solution so you're not looking for a needle in a haystack, (or hockey puck!) while sectioning.

Hope this helps!
--
Bradford Ross

Microscopy Technician
BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996




-----Original Message-----

} Date: Wed Nov 17 16:30:12 PST 2010
} From: gw265-at-cam.ac.uk
} Subject: [Microscopy] chasing small specimens round the AFS
} To: bnross-at-interchange.ubc.ca
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} Does anyone have any advice on how to not lose tiny specimens during
} freeze substitution? They're currently sitting in the planchettes (hats)
} in which they were high pressure frozen, at -90 in acetone + glut + tannic
} acid.
}
} While the specimens are below zero, ok, they stay in the planchette and
} you can change solutions easily enough. But we're wondering if they fall
} out once things go above zero, and if so, then what do you do? Is
} trapping in agar an option? Seems a bad idea to go from osmium + acetone,
} to acetone, to agar + water, back to acetone.... but if that's the only
} way to hang onto the specimen, we'll do it.
}
} When doing chemical fixations at room temperature we would traditionally
} fix cells in solution, pellet, resuspend in washes, and eventually trap in
} agar, then dehydrate the agar.
}
} Any advice greatly appreciated - this part of the process has always been
} entrusted to experienced techs before, and I've been stupid enough to fail
} to ask what they did. The techs where I'm working have only worked with
} bigger bits of tissue!
}
} thanks
}
} Giselle
}
} Dr Giselle Walker
} University of Cambridge
}
}
} ==============================Original Headers==============================
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--
Bradford Ross

Microscopy Technician
BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996


==============================Original Headers==============================
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From: varguc-at-freemail.hu
Date: Tue, 23 Nov 2010 14:15:02 -0600
Subject: [Microscopy] Problems with an old LINK AN10/55 system

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear List,

we need your help. We have an old LINK AN10/55S system kept in good working conditions up to now. However, recently something has gone wrong and the instrument cannot be operated any more. Some time ago we had some keyboard problems. Some keys were unoperable but the insrument could be used with some care. Then an expert fixed the keys and everything was OK for some time. Now the instrument cannot be started. When we turn it on, the usual few lines of information appear on the screen and the instrument waits forever for the Enter key to be pressed, probably due to an interconnection
fault between the keyboard and the processor. The printer does not work, either.Both of them seem to get signals from the same board. We asked some help from Nano Analysis Hotline. The main point of their answer is: "One thing that can cause the type of fault you are seeing is the hard drive can get stuck and you should take it out and twist the spindle". (???)
The hard drive is a 20 Mbytes one with a SCSI interface (as far as I can remember). More than 15 years ago the hard drive once was overwritten. The Demon operating system seems to have no protection against it so we had to format the drive and install everything again.
Your experience, suggestions, spare parts (keyboard, hard drive etc.) any kind of help would be highly appreciated. Thank you in advance:

Mt. Laszlo Varga,
computer programmer (a former microscopist)

==============================Original Headers==============================
1, 16 -- From varguc-at-freemail.hu Tue Nov 23 14:15:02 2010
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1, 16 -- From: Varga Laszlo {varguc-at-freemail.hu}
1, 16 -- Subject: [Microscopy] Problems with an old LINK AN10/55 system
1, 16 -- To: Microscopy {Microscopy-at-microscopy.com}
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From: ron.doole-at-materials.ox.ac.uk
Date: Tue, 23 Nov 2010 15:10:23 -0600
Subject: [Microscopy] Problems with an old LINK AN10/55 system

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Laszlo,

When we were running our AN10 we had problems restarting if it was shut off for any period of time, the problem was the hard disc and the cure was to manually spin it and then switch it on, as the helpline have suggested. I guess this problem started after about 8-10 years of almost continual running, it seems that the bearing grease dries out and sticks when it gets cold. If you have not given it a try do so it has a good chance of working.

Regards,
Ron

Mr. Ron Doole Department of Materials
Senior Instrumentation Engineer. University of Oxford.
Phone +44 (0) 1865 273701 Parks Road. Oxford. OX1 3PH
Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
________________________________________
X-from: varguc-at-freemail.hu [varguc-at-freemail.hu]
Sent: 23 November 2010 20:28
To: Ron Doole

Dear List,

we need your help. We have an old LINK AN10/55S system kept in good working conditions up to now. However, recently something has gone wrong and the instrument cannot be operated any more. Some time ago we had some keyboard problems. Some keys were unoperable but the insrument could be used with some care. Then an expert fixed the keys and everything was OK for some time. Now the instrument cannot be started. When we turn it on, the usual few lines of information appear on the screen and the instrument waits forever for the Enter key to be pressed, probably due to an interconnection
fault between the keyboard and the processor. The printer does not work, either.Both of them seem to get signals from the same board. We asked some help from Nano Analysis Hotline. The main point of their answer is: "One thing that can cause the type of fault you are seeing is the hard drive can get stuck and you should take it out and twist the spindle". (???)
The hard drive is a 20 Mbytes one with a SCSI interface (as far as I can remember). More than 15 years ago the hard drive once was overwritten. The Demon operating system seems to have no protection against it so we had to format the drive and install everything again.
Your experience, suggestions, spare parts (keyboard, hard drive etc.) any kind of help would be highly appreciated. Thank you in advance:

Mt. Laszlo Varga,
computer programmer (a former microscopist)

==============================Original Headers==============================
1, 16 -- From varguc-at-freemail.hu Tue Nov 23 14:15:02 2010
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1, 16 -- Date: Tue, 23 Nov 2010 21:14:14 +0100 (CET)
1, 16 -- From: Varga Laszlo {varguc-at-freemail.hu}
1, 16 -- Subject: [Microscopy] Problems with an old LINK AN10/55 system
1, 16 -- To: Microscopy {Microscopy-at-microscopy.com}
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==============================Original Headers==============================
9, 30 -- From ron.doole-at-materials.ox.ac.uk Tue Nov 23 15:10:22 2010
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9, 30 -- From: Ron Doole {ron.doole-at-materials.ox.ac.uk}
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9, 30 -- Date: Tue, 23 Nov 2010 21:10:20 +0000
9, 30 -- Subject: RE: [Microscopy] Problems with an old LINK AN10/55 system
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From: wesaia-at-iastate.edu
Date: Tue, 23 Nov 2010 15:53:01 -0600
Subject: [Microscopy] Problems with an old LINK AN10/55 system

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I have not had experience with an AN10, but I had similar troubles with an old personal computer. I concur with your diagnosis.

I normally left the computer run around the clock. It stopped running during a power failure and the disk cooled off. Supposing that the lubricant had gotten stiff, I removed the disk and put it into a warm place (50-70C) for about 30 minutes. I removed it, reinstalled it, and it booted up fine.

I then replaced the drive as quickly as I could.

ws
________________________________________
X-from: ron.doole-at-materials.ox.ac.uk [ron.doole-at-materials.ox.ac.uk]
Sent: Tuesday, November 23, 2010 3:11 PM
To: wesaia-at-iastate.edu

Dear Laszlo,

When we were running our AN10 we had problems restarting if it was shut off for any period of time, the problem was the hard disc and the cure was to manually spin it and then switch it on, as the helpline have suggested. I guess this problem started after about 8-10 years of almost continual running, it seems that the bearing grease dries out and sticks when it gets cold. If you have not given it a try do so it has a good chance of working.

Regards,
Ron

Mr. Ron Doole Department of Materials
Senior Instrumentation Engineer. University of Oxford.
Phone +44 (0) 1865 273701 Parks Road. Oxford. OX1 3PH
Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
________________________________________
X-from: varguc-at-freemail.hu [varguc-at-freemail.hu]
Sent: 23 November 2010 20:28
To: Ron Doole

Dear List,

we need your help. We have an old LINK AN10/55S system kept in good working conditions up to now. However, recently something has gone wrong and the instrument cannot be operated any more. Some time ago we had some keyboard problems. Some keys were unoperable but the insrument could be used with some care. Then an expert fixed the keys and everything was OK for some time. Now the instrument cannot be started. When we turn it on, the usual few lines of information appear on the screen and the instrument waits forever for the Enter key to be pressed, probably due to an interconnection
fault between the keyboard and the processor. The printer does not work, either.Both of them seem to get signals from the same board. We asked some help from Nano Analysis Hotline. The main point of their answer is: "One thing that can cause the type of fault you are seeing is the hard drive can get stuck and you should take it out and twist the spindle". (???)
The hard drive is a 20 Mbytes one with a SCSI interface (as far as I can remember). More than 15 years ago the hard drive once was overwritten. The Demon operating system seems to have no protection against it so we had to format the drive and install everything again.
Your experience, suggestions, spare parts (keyboard, hard drive etc.) any kind of help would be highly appreciated. Thank you in advance:

Mt. Laszlo Varga,
computer programmer (a former microscopist)

==============================Original Headers==============================
1, 16 -- From varguc-at-freemail.hu Tue Nov 23 14:15:02 2010
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1, 16 -- From: Varga Laszlo {varguc-at-freemail.hu}
1, 16 -- Subject: [Microscopy] Problems with an old LINK AN10/55 system
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==============================Original Headers==============================
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==============================Original Headers==============================
16, 37 -- From wesaia-at-iastate.edu Tue Nov 23 15:53:01 2010
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16, 37 -- "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
16, 37 -- Date: Tue, 23 Nov 2010 15:52:56 -0600
16, 37 -- Subject: RE: [Microscopy] RE: Problems with an old LINK AN10/55 system
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From: r-bleher-at-northwestern.edu
Date: Tue, 23 Nov 2010 20:12:34 -0600
Subject: [Microscopy] viaWWW: Open Position: EM Technician at Northwestern University

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Email: r-bleher-at-northwestern.edu
Name: Reiner BLeher

Organization: Northwestern University

Title-Subject: [Filtered] Open Position: EM Technician at
Northwestern University

Message: Technician: Biological Specimen Preparation

The Keck Inorganic Signatures of Life Program in collaboration with
QBIC (Quantitative Biological Imaging Center:
http://chemgroups.northwestern.edu/qbic/ and the NUANCE (Northwestern
University's Atomic and Nanoscale Characterization Experimental
Center: http://www.nuance.northwestern.edu/epic at Northwestern
University have an immediate opening for a technician with background
in biological- and cryo-biological specimen preparation for electron
microscopy and related techniques.
The main tasks for this position include preparation of biological
samples for S/T/EM with conventional methods and cryo-techniques,
including plunge freezing and high pressure freezing, (cryo)
ultramicrotomy of samples, and observation of specimens with (cryo)
S/TEM and SEM. There are plenty opportunities for personal and
professional growth in the dynamic and vibrant research environment
of QBIC/NUANCE and Northwestern University.
The position requires hands-on experience in specimen preparation for
biological and soft materials, and basic knowledge in related
sciences and/or engineering themes.
Applications including a detailed CV, a list of three references with
email addresses and telephone numbers, and salary requirements should
be sent electronically using the following link:
http://www.northwestern.edu/hr/jobs/. After registering, use NUANCE
as keyword for job search. The offer ID is 16480. Only applications
sent through the eRecruit system will be considered.
Northwestern University is an equal opportunity, affirmative action employer.
Members of historically underrepresented groups are strongly
encouraged to apply.

Reiner Bleher
Northwestern University


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From: gmf.decaro-at-tiscali.it
Date: Tue, 23 Nov 2010 20:13:00 -0600
Subject: [Microscopy] viaWWW: JENAVAL MICROSCOPE - MANUAL NEEDED

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Email: gmf.decaro-at-tiscali.it
Name: Giovanni De Caro, MD

Organization: Ospedale Cardarelli - Campobasso - Italy

Title-Subject: [Filtered] JENAVAL MICROSCOPE - MANUAL NEEDED

Message: I urge the manual of the Carl Zeiss Jena JENAVAL research
microscope. Anyone of you can provide me one in pdf. format or send
me a photocopy (I pay for postal fee)? Please let me know. The manual
must be in english. Thank you.

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From: rpowell-at-nanoprobes.com
Date: Tue, 23 Nov 2010 20:13:43 -0600
Subject: [Microscopy] viaWWW: Re: FEG-SEM High Vacuum, High Current, WDS Plant Analysis

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Email: rpowell-at-nanoprobes.com
Name: Rick Powell

Organization: Nanoprobes, Incorporated

Title-Subject: [Filtered] Re: FEG-SEM High Vacuum, High Current, WDS
Plant Analysis

Message: [Commercial disclosure - we make silver enhancement reagents]

Hello Ivan:

Have you considered using autometallography to amplify the signal
from the metals? Although this is more familiar when used with
immunogold, Gorm Danscher has published a number of papers where he
uses treatment with metals and a reducing agent to demonstrate the
presence of traces of a number of different metals in biological
systems, including a couple of references that are specific for zinc.

You might check these references:

(1) Danscher, G., and Stoltenberg, M.: Zinc-specific
Autometallographic In Vivo Selenium Methods: Tracing of Zinc-enriched
(ZEN) Terminals, ZEN Pathways, and Pools of Zinc Ions in a Multitude
of Other ZEN Cells. J. Histochem. Cytochem., 53141-153 (2005).

(2) Danscher, G., and Stoltenberg, M.: Silver enhancement of quantum
dots resulting from (1) metabolism of toxic metals in animals and
humans, (2) in vivo, in vitro and immersion created
zinc-sulphur/zinc-selenium nanocrystals, (3) metal ions liberated
from metal implants and particles. Prog. Histochem. Cytochem., 41,
57-139 (2006).

(3) Stoltenberg, M., and Danscher, G.: Histochemical differentiation
of autometallographically traceable metals (Au, Ag, Hg, Bi, Zn):
protocols for chemical removal of separate Autometallographic metal
clusters in Epon sections. Histochem. J., 32, 645-52 (2000).

Not sure if these are applicable to your system, but hope this is some help,

Rick Powell


*****************************************************************************************
Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com

NANOPROBES, Incorporated
*****************************************************************************************

Hello,
I am looking for some suggestions of how to run some FEG-SEM/WDS
analyses on plant material.
I am a geologist by trade, and this will be the first time I've run
plant material on the SEM so I'm seeking some helpful tips from some
more experienced analysts.
We have a Hitachi SU6600 FEG-SEM with a WDS detector. We are trying
to determine the distribution of Cd and Zn in plant root structures,
even though the Cd and Zn are in very low concentrations.
We would like to map the Cd and Zn using WDS. However, since they are
at such low concentrations, I believe I would need very high probe
current and long mapping times, which would easily destroy the plant
material. I was thinking of running the analysis at 20kV, 150nA, and
under high vacuum (the WDS detector does not work in VP mode).
How have people over come sample damage for this type of analysis? Or
have you done this type of analysis in a different way? How do you
prep the samples (inject with conductive epoxy?), what do you coat
with, and what would be the best operating conditions?
Please respond to ibarke2-at-uwo.ca
Many thanks!



Ivan R. Barker
UWO Earth Sciences MSc
ZAPLab Geochronology Research Technician
T: 519-661-2111 ext. 88397
E: ibarke2-at-uwo.ca

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From: kwylly-at-ppg.com
Date: Tue, 23 Nov 2010 20:14:12 -0600
Subject: [Microscopy] viaWWW: SAICAS Testing

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Email: kwylly-at-ppg.com
Name: Kelly Wylly

Organization: PPG Industries

Title-Subject: [Filtered] SAICAS Testing

Message: This may be outside the scope of microscopy, but have any of
you heard of SAICAS testing that involves using a microtome to
measure the peel strength at different coating interfaces?

If you know anything about this testing or where this testing could
be performed, can you please let me know?

Thanks!

Kelly Wylly

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From: M.A.E.Jepson-at-lboro.ac.uk
Date: Wed, 24 Nov 2010 03:53:25 -0600
Subject: [Microscopy] electro-thinning Ni-based superalloy, CMSX4

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Dear all,

Firstly my apologies for posting this message twice!

I have been attempting to electro-thin 3mm discs of CMSX4 for TEM examination but am having some trouble obtaining useful samples. I am using a Struers Tenupol-5 with an etchant of 5% perchloric acid in methanol.

I have tried many many voltages, sample thicknesses and electrolyte temperatures and from the 50 or discs I have tried, only 1 is of any use! I have a feeling I may be using the wrong electrolyte for this application. Does anybody have any experience of thinning this material? If so, could you please advise me which electrolyte to use and perhaps suggest some starting conditions.... Any help which anyone can give will be very much appreciated.

Many thanks and regards

Mark Jepson

Dr Mark A. E. Jepson BEng (Hons), PhD

Department of Materials,
Loughborough University,
Loughborough,
Leicestershire.
LE11 3TU

Tel: +44 (0)1509 223169
http://www.lboro.ac.uk/departments/materials



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From: m.a.e.jepson-at-lboro.ac.uk
Date: Wed, 24 Nov 2010 08:26:45 -0600
Subject: [Microscopy] viaWWW: Electro-thinning Ni-based superalloy (CMSX4)

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Email: m.a.e.jepson-at-lboro.ac.uk
Name: Mark Jepson

Organization: Loughborough University, UK

Title-Subject: [Filtered] Electro-thinning Ni-based superalloy (CMSX4)

Message: Dear all,

I have been attempting to electro-thin 3mm discs of CMSX4 for TEM
examination but am having some trouble obtaining useful samples.

I am using a Struers Tenupol-5 with an etchant of 5% perchloric acid
in methanol. i have tried many many voltages, sample thicknesses and
electrolyte temperatures and from the 50 or discs I have tried, only
1 is of any use! I have a feeling I may be using the wrong
electrolyte for this application.

Does anybody have any experiece of thinning this material? If so,
could you please advise me which electrolyte to use and perhaps
suggest some starting conditions....

Any help which anyone can give will be very much appreciated.


Many thanks and regards

Mark Jepson

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From: bzuber-at-mrc-lmb.cam.ac.uk
Date: Wed, 24 Nov 2010 10:57:19 -0600
Subject: [Microscopy] 1st UB Practical course on Cryo-Electron Microscopy of Vitreous

Contents Retrieved from Microscopy Listserver Archives
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We are pleased to announce the

1st UB Practical course on Cryo-Electron Microscopy of Vitreous Sections
(CEMOVIS)
20-24 June 2011
University of Bern, Switzerland

Organisers : Daniel Studer & Benoît Zuber
Invited speakers: Jacques Dubochet (University of Lausanne), Helmut Gnaegi
(Diatome), Bruno Humbel (University of Lausanne).

The objective of the course is that participants get the practical skills
necessary to successfully apply CEMOVIS back in their laboratories.
Background theory on various aspects of CEMOVIS will be given by the
organisers and the invited speakers. Most of the time will be spent
practicing high-pressure freezing, cryo-sectioning, and low-dose TEM
imaging.

The course is intended for scientists whose research projects will benefit
from the use of CEMOVIS. Experience in either cryo-electron microscopy or
ultramicrotomy of resin-embedded specimens is a prerequisite. The number
of participants will be very limited to ensure optimal training.
Interested candidates are therefore invited to register early.

A registration fee applies and includes lunch. (PhD students: CHF 400;
non-profit organisation employees: CHF 800; others: CHF 1600).

To register please send a curriculum vitae and a cover letter briefly
describing your current or future research and how it is going to benefit
from CEMOVIS to daniel.studer-at-ana.unibe.ch or bzuber-at-mrc-lmb.cam.ac.uk.

Daniel Studer & Benoît Zuber


-----------
Benoît Zuber
MRC Laboratory of Molecular Biology
Hills Road
Cambridge
CB2 0QH
UK


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From: bnross-at-interchange.ubc.ca
Date: Wed, 24 Nov 2010 12:56:24 -0600
Subject: [Microscopy] chasing small specimens round the AFS

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi Allan,

You're welcome. I don't believe we've made the 20% BSA solution with culture media, but I know we've used either buffer or water, depending on the needs of the sample.

Thanks,
-Brad


-----Original Message-----

} Date: Tue Nov 23 10:58:40 PST 2010
} From: "Allan Mitchell" {allan.mitchell-at-stonebow.otago.ac.nz}
} Subject: Re: [Microscopy] Re: chasing small specimens round the AFS
} To: bnross-at-interchange.ubc.ca
}
} Hi Brad
}
} Great tip, thanks.
}
} Is the BSA made up in buffer, cell media or water?
}
}
} Allan
}
}
} On 24/11/2010, at 7:56 AM, {bnross-at-interchange.ubc.ca} wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } Hi Giselle,
} }
} } When we have to do high pressure freezing with cells in suspension, we usually mix them in a 20% solution of BSA just before freezing. The BSA reacts with Osmium in the freeze-substitution process and most of the time you end up with a nice little solid brown/black "hockey puck" that is chock full of cells and very easy to embed and section. When you mix the cells with the BSA, try to have the cells as concentrated as possible in the BSA solution so you're not looking for a needle in a haystack, (or hockey puck!) while sectioning.
} }
} } Hope this helps!
} } --
} } Bradford Ross
} }
} } Microscopy Technician
} } BioImaging Facility
} } University of British Columbia
} } 6270 University Blvd.
} } Vancouver, B.C.
} } Canada
} } V6T 1Z4
} }
} } phone 604-822-6996
} }
}
--
Bradford Ross

Microscopy Technician
BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996


==============================Original Headers==============================
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9, 27 -- From: Bradford Ross {bnross-at-interchange.ubc.ca}
9, 27 -- Subject: Re: [Microscopy] Re: chasing small specimens round the AFS
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From: bnross-at-interchange.ubc.ca
Date: Wed, 24 Nov 2010 16:05:21 -0600
Subject: [Microscopy] RE: chasing small specimens round the AFS

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Rick,

Thanks for the tips! We usually do a .5% Glut/.1% Tannic Acid intermediate step in our FS for cultured cells, or anything else that benefits from enhanced membrane contrast. Could this be making our pucks a bit darker than yours? They usually range from light brown to black, and I don't see any problems with the ultrastructure unless something has gone obviously wrong, such as ice crystal damage.

Thanks,
-Brad


-----Original Message-----

} Date: Wed Nov 24 11:34:22 PST 2010
} From: "Fetter, Richard" {fetterr-at-janelia.hhmi.org}
} Subject: RE: [Microscopy] chasing small specimens round the AFS
} To: "bnross-at-interchange.ubc.ca" {bnross-at-interchange.ubc.ca}
}
} Brad,
}
} The BSA can be made up in medium used for cultured cells, whether in suspension or grown on sapphire discs for HPF. We do this routinely with excellent results since the 20% BSA doesn't add significant osmotic strength to the solution.
}
} After OsO4 FS, the BSA should *not* turn dark brown or black. If it does, it indicates contamination in the acetone or other organic solvent being used. In the past, I used 'Molecular Sieves' to 'dry out' my acetone. I got the black pucks. When using the acetone direct from the reagent bottle, no black pucks, and better looking cells.
}
} After FS in OsO4 in acetone, the BSA puck should be a light tan in color.
}
} Rick
}
} -----Original Message-----
} From: bnross-at-interchange.ubc.ca [mailto:bnross-at-interchange.ubc.ca]
} Sent: Wednesday, November 24, 2010 2:07 PM
} To: Fetter, Richard
} Subject: [Microscopy] chasing small specimens round the AFS
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi Allan,
}
} You're welcome. I don't believe we've made the 20% BSA solution with culture media, but I know we've used either buffer or water, depending on the needs of the sample.
}
} Thanks,
} -Brad
}
}
} -----Original Message-----
}
} } Date: Tue Nov 23 10:58:40 PST 2010
} } From: "Allan Mitchell" {allan.mitchell-at-stonebow.otago.ac.nz}
} } Subject: Re: [Microscopy] Re: chasing small specimens round the AFS
} } To: bnross-at-interchange.ubc.ca
} }
} } Hi Brad
} }
} } Great tip, thanks.
} }
} } Is the BSA made up in buffer, cell media or water?
} }
} }
} } Allan
} }
} }
} } On 24/11/2010, at 7:56 AM, {bnross-at-interchange.ubc.ca} wrote:
} }
} } }
} } }
} } }
} } } ----------------------------------------------------------------------------
} } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } } ----------------------------------------------------------------------------
} } }
} } } Hi Giselle,
} } }
} } } When we have to do high pressure freezing with cells in suspension, we usually mix them in a 20% solution of BSA just before freezing. The BSA reacts with Osmium in the freeze-substitution process and most of the time you end up with a nice little solid brown/black "hockey puck" that is chock full of cells and very easy to embed and section. When you mix the cells with the BSA, try to have the cells as concentrated as possible in the BSA solution so you're not looking for a needle in a haystack, (or hockey puck!) while sectioning.
} } }
} } } Hope this helps!
} } } --
} } } Bradford Ross
} } }
} } } Microscopy Technician
} } } BioImaging Facility
} } } University of British Columbia
} } } 6270 University Blvd.
} } } Vancouver, B.C.
} } } Canada
} } } V6T 1Z4
} } }
} } } phone 604-822-6996
} } }
} }
} --
} Bradford Ross
}
} Microscopy Technician
} BioImaging Facility
} University of British Columbia
} 6270 University Blvd.
} Vancouver, B.C.
} Canada
} V6T 1Z4
}
} phone 604-822-6996
}
}
} ==============================Original Headers==============================
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--
Bradford Ross

Microscopy Technician
BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996


==============================Original Headers==============================
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From: drg.mitchell-at-sydney.edu.au
Date: Wed, 24 Nov 2010 17:10:51 -0600
Subject: [Microscopy] Re: electro-thinning Ni-based superalloy, CMSX4

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Michael

I use 4% perchloric in methanol at -40C in a Tenupol. It it my universal
polishing solution and I have had great results with everything from mild
steel, austenitics, superalloys, rare earths, titanium etc.

The trick with electropolishing is to forget about voltage and concentrate
on current. It is current which controls the electrochemical dissolution,
the voltage will vary all over the place as the conductivity of the solution
changes with increasing use (more metal salts). I always discard my solution
after use. Metal perchlorate salts, which crystallise out on bottle
stoppers, can be explosive.

For double-side thinning I use a current of between 120 -200mA. Start at the
low end and increase the current in 20mA increments for subsequent
specimens. Examine them to see where the sweet spot is. If you are right on
the edge of good electropolishing conditions, you might find what works one
day, doesn't the next.

If your current is too low, you will find your specimens to be badly
oxidised. If it is too high, your specimens will perforate very quickly
( {20s) and you have pinholes which look like apertures. This is based on
specimens ground to around 80ums thick. If you are working with
ferromagnetics, you might want to grind a little thinner, to reduce the
effect on the TEM fields. Electropolishing times will typically be between
20 and 60s.

Some other things to think about. Flow rate is not too critical, I use 3 on
a scale of 1 (slow) to 10 (fast). Temperature is very important. Low
temperatures reduce oxidation and increase viscosity, favouring the
establishment of good electopolishing conditions. You didn't mention if you
were cooling. If not it is a must with this solution. A small dewar, some
silicone tubing and a diaphragm pump to draw the LN2 through the Tenupol's
cooling loop and you're in business.

Cleanliness is paramount. Clean all equipment with methanol before and after
use. Make sure there is no water contamination in any of your
solutions/equipment. Set the optical sensitivity to maximum, and get the
specimen out of the Tenupol and into a beaker of methanol immediately the
perforation alarms sounds. When electropolishing stops - corrosion starts
and you might get etch pitting. Wash your specimens very, very thoroughly
with pure methanol from a wash bottle and air dry - do not use acetone.
Store specimens in a desiccator, never under vacuum.

I place all perchloric acid contaminated tissue waste into a very large
beaker of water as I go. At the end I rinse this through with fresh water,
then dispose of it sealed inside a ziplock bag, so there's no danger of
spontaneous combustion.

Good luck.

Regards

Dave Mitchell


Dr David Mitchell
Senior Microscopist, Laboratory Manager (Acting)

Contact:
PH +61 2 9036 7633
FAX +61 2 9351 7682
drg.mitchell-at-sydney.edu.au

Address:
Australian Centre for Microscopy & Microanalysis
Incorporating:
Australian Microscopy & Microanalysis Research Facility
ARC Centre of Excellence for Design in Light Metals
Madsen Building F09, Room 128A
The University of Sydney
NSW, 2006, Australia
sydney.edu.au/acmm {http://www.sydney.edu.au/acmm}
www.ammrf.org.au {http://www.ammrf.org.au/}

www.dmscripting.com



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16, 27 -- Subject: Re: [Microscopy] electro-thinning Ni-based superalloy, CMSX4
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From: allan.mitchell-at-stonebow.otago.ac.nz
Date: Thu, 25 Nov 2010 19:35:18 -0600
Subject: [Microscopy] EM Unit User Charges

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi all,

We are about to review our Business Plan and User Charges. With our current business plan we budget to recover all our costs. This includes: floor space rental, electricity, water, cleaning, staff salaries, all equipment maintenance and repair including the electron microscopes, replacement of preparation equipment (from pH meter to ultramicrotomes) and consumables. The only thing we do not recover is depreciation on the major equipment. Our University deemed this as unsustainable and would put research and teaching costs out of reach.

Our current funding model is a Club Model. Departments choose either to join the Club or not join. If they join then their researchers and students get a reduced rate on the equipment usage and don't pay for our technical support time. If a department does not join the Club then researchers and students from that department pay a higher equipment usage charge and also pay for our technical support time. In either case the consumable costs are the same.

To join the Club a department must pay an annual fee. This fee is calculated on the proportion of time that their researchers used the EM Unit in the last three year period (a 3 year rolling average is calculated each year). This is a subsidy from a department for their students and researchers.

If a department does not join the Club then each individual researcher and student from that department pays a equipment usage rate that is approximately two-thirds higher than the Club rate. They also pay our labour. All the club fees combined cover two-thirds of the Units annual costs (less consumables). The remaining third is recovered from the reduced rate equipment charges to club members , or the higher rate equipment charges to non-club members and labour charges, or any external work we do.

There is no underwriting from the host institution, we are expected to break even each year.

This system has by and large worked well for the last couple of years. On the one hand I have been able to budget each year for things that otherwise we would have had to leap through flaming hoops for. It has made EM more expensive for smaller departments or individual researchers from non-Club member departments. It has reduced the opportunity to experiment with techniques and to run pilot studies. We have noted a fall off in undergraduate exposure and we are getting postgraduates coming in who have been told how many hours they can have, including training. I don't believe the students are being well served by this (getting the wrong idea of EM, expensive to do rather than appreciating the value of the data they collect) and I think we are seeing a fall off in quality.

I have been asked to do a survey by by User Group to find out

a) if other EM Centres have to recover all of their costs (overhead and operational), or just operational costs and overhead costs are underwritten,

b) if full cost recover (overheads and operational) , what funding model is used?

c) what charge out differentials are applied for students, pilot studies, high usage, etc?

We are hoping to move to a model where pilot studies can be undertaken at minimal cost, higher usage attracts a lower charge, and postgraduate students are not penalised because they come from 'poorer' department,or non-club department.

All comments and feedback appreciated.

Have a great weekend

Allan




Allan Mitchell
Microscopy Otago - Electron Microscopy
c/- Department of Anatomy and Structural Biology
Otago School of Medical Sciences
P.O. Box 913
Dunedin
New Zealand

Phone (03) 479 5642 or 479 7301
Fax (03) 479 5086 or 479 7254

EM Centre: http://ocem.otago.ac.nz/
Confocal Centre: http://occm.otago.ac.nz/
NZ Microscopy Society: http://microscopynz.co.nz/





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From: dcd-at-unimelb.edu.au
Date: Mon, 29 Nov 2010 09:07:38 -0600
Subject: [Microscopy] viaWWW: Post-staining with triple lead

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Email: dcd-at-unimelb.edu.au
Name: Chaitali Dekiwadia

Organization: Melbourne university

Title-Subject: [Filtered] Post-staining with triple lead

Message:
Hi,

I post-stain the immunolabeled samples with triple lead. I try to
avoid precipitate formation using NaOH pellets around the staining
chamber, however there is lot of precipitation on the grids even if i
wash them several times in Milli Q water.

Is it advisable to wash the grids in 0.1N NaOH solution and re-stain
them again ?
If i wash them in NaOH solution will it affect the immunolabeling reaction?

Regards,
Chaitali

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From: PhillipsT-at-missouri.edu
Date: Mon, 29 Nov 2010 10:26:44 -0600
Subject: [Microscopy] viaWWW: Post-staining with triple lead

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Chaitali,

We seldom use lead stain on immuno. sections but rather use a smaller
Objective Aperture to boost tissue contrast, at least for initial viewing.

Whenever there is precipitation it is always good to look at an unstained
section to check if the precipitation is in the section rather than on it as
a result of the staining procedure. After checking that it is fine you can
stain the same grid for imaging if you wish.

Did you syringe-filter the stain before use or at least allow the stain to
settle a while before taking the stain from near the top of the tube?

I have also found less frequent lead precipitation on sections if I have the
grids submerged in non-acid water drops (1 pellet of NaOH in 0.5 L) before
putting them into the lead stain drops. I have also used water from this
same bottle for a brief rinse after the lead stain followed by some good
water washes.

I have not tried to wash off precipitation recently for I now pick up more
grids than I need ­ usually five. That way if I stain two, I have three in
reserve if there is a need to check or correct a staining problem. Many
years ago I did attempt to wash off lead precipitation but I must not have
done it correctly since it did not work for me and I have not done it since.

Hopefully others will add to my comments especially to the point of washing
the grids in 0.1N NaOH solution.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E ­ Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and can be freely shared
and reproduced.
============

X-from: {dcd-at-unimelb.edu.au}
Reply-To: {dcd-at-unimelb.edu.au}

My understanding is that to remove lead precipitate from sections, it is necessary to incubate the sections on an acidic solution. This is why lead staining has to follow uranyl acetate staining since the acidic uranyl solution would remove the lead. My guess is some precipitates will be refractory to being re-dissolved and removed but it is worth a try. My recollection is that once you have viewed a grid in the TEM, it is especially difficult, if not impossible, to remove precipitates so this is really only an option if you stained numerous grids and haven't looked at them all. Good luck. Tom




Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


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Email: dcd-at-unimelb.edu.au
Name: Chaitali Dekiwadia

Organization: Melbourne university

Title-Subject: [Filtered] Post-staining with triple lead

Message:
Hi,

I post-stain the immunolabeled samples with triple lead. I try to
avoid precipitate formation using NaOH pellets around the staining
chamber, however there is lot of precipitation on the grids even if i
wash them several times in Milli Q water.

Is it advisable to wash the grids in 0.1N NaOH solution and re-stain
them again ?
If i wash them in NaOH solution will it affect the immunolabeling reaction?

Regards,
Chaitali

Login Host: 60.228.112.27
---------------------------------------------------------------------------

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From: sgkcck-at-aol.com
Date: Mon, 29 Nov 2010 19:14:59 -0600
Subject: [Microscopy] viaWWW: Aurion Immuno Gold Silver Staining Workshop

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Email: sgkcck-at-aol.com
Name: Stacie Kirsch

Organization: Electron Microscopy Sciences

Title-Subject: [Filtered] Aurion Immuno Gold Silver Staining Workshop

Message: Aurion Immuno Gold Silver Staining Workshop
House Ear Institute
Los Angeles, California
February 21-23, 2011

The objective of the course is to provide researchers with the
opportunity to learn the theory and practice of immunogold labeling.
Participants will process their own samples under the expert guidance
of Mr. Peter van de Plas and Dr. Paul Webster. Mr van de Plas has
been with Aurion since 1991 and has been involved with product
applications and workshops, and is recognized for his through
knowledge of the immunogold silver staining techniques. Dr. Webster
is head of Ahmanson Advanced EM & Imaging Center at the at the House
Ear Institute and is widely known for his expertise in immunolabeling
and applying electron microscopy to biomedical research problems.

The brochure is available online
http://www.emsdiasum.com/microscopy/products/immunogold/workshop.pdf.

Please contact Stacie Kirsch for more information.

Tel: 215-412-8400 Fax: 215-412-8450
E-Mail: stacie-at-ems-secure.com, sgkcck-at-aol.com

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From: hyi-at-emory.edu
Date: Tue, 30 Nov 2010 09:40:38 -0600
Subject: [Microscopy] Contrast of cryoultrathin sections

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Dear EM Colleagues:

Hope you all have had a good holiday weekend.

I have been using two methods (See below) for staining/drying
cryo-ultrathin sections (by Tokuyasu method), but was not able to get very
good contrast. Does anyone have suggestions for what other methods I should
try? Thank you very much in advance.

Method 1:

Mix nine parts of 2% aqueous methyl cellulose with one part 4% aqueous
uranyl acetate solution. MC/UA mixture on ice for 10 minutes. Loop off grids
and drag the edge of the loop across a sheet of filter paper. Air dry.

Method 2:

Mix equal parts of 0.3 M oxalic acid with 4% aqueous uranyl acetate and
bringing the pH to about 7.5 with NH4OH. (Use pH paper). Float the grids,
section side down, on drops of UA/OA mixture for 5 minutes. Wash sections
with DD water, and then continue as described in Method 1.


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From: jpennin-at-purdue.edu
Date: Tue, 30 Nov 2010 10:46:44 -0600
Subject: [Microscopy] Contrast of cryoultrathin sections

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Hong Yi,
In my hands the classic 9 parts 2% methyl cellose to 1 pt. 4% UA has always
resulted in what I call positive staining. A slight increase in the
concetration of UA will make a world of difference. A lower percentage of
uranyl acetate results in a positive stain and a higher percentage results in a
negative stain. And then there is the grey area in-between when it may stain
positive or negative as is often seen with negative staining with viruses.

As you know the Tokuyasu technique has classically been used as a technique for
immunolabeling of membranes. This technique allows membranes in tissue to be
processed without the destructive effect that alchohol has on lipids. When the
tissue is truely negatively stained the only thing you will see is the white
outline of membranes.

When I first started a project using this method to look at golgi, I could not
see any golgi! When I started playing around with different concetrations I
finally came up with something that worked for me and gave beautiful negative
stain and beautiful ribbons of golgi.

I have not had the opportunity to do the Tokuyasu technique for several years
so I am having to dust the cobwebbs from my brain. If I look through my notes
I may be able to give you more specifics. However, the best way is to just
play with different concentrations with out lowering the concentration of MC
too much. I think I have gone as low as 1.5% MC (2% diluted with the UA).

My 2 cents!

Janice G. Pennington, M.S.
Research Electron Microscpist
Biological Electron Microscopy Facility
Department of Biological Sciences
Purdue University
jpennin-at-purdue.edu
(765)496-6781




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From: jehrman-at-mta.ca
Date: Tue, 30 Nov 2010 11:41:47 -0600
Subject: [Microscopy] for the microscopist who has everything...

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Season's Greetings all,

I was casting a wide net for lab gifts and came across this:

http://www.cafepress.com/+pay_electron_microscope_mug,340868475

or for all of us GWNers:

http://www.cafepress.ca/+pay_electron_microscope_mug,340868475

They have other microscopy related things, too (search for "electron
microscope") but this is my favorite!

No interest in the company, just several years of amusing and satisfying
gifts for all the people who suffer our particular flavor of craziness!

Jim

--

James M. Ehrman
Digital Microscopy Facility
Mount Allison University
63B York St.
Sackville, NB E4L 1G7
CANADA

phone: 506-364-2519
fax: 506-364-2505
email: jehrman-at-mta.ca
www: http://www.mta.ca/dmf

I doubt therefore I might be.


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From: Marta.Jarquin-Pardo-at-STJUDE.ORG
Date: Tue, 30 Nov 2010 12:46:01 -0600
Subject: [Microscopy] Director of light microscopy - St Jude Children's Research hospital

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Director of Light Microscopy - Cell and Tissue Imaging Center
St. Jude Children's Research Hospital
(Job Number 18238)

We announce an exciting opportunity for experts in live cell imaging technology to direct a high profile facility advancing research in cancer cell biology. The Cell and Tissue Imaging Center (CTIC) at St. Jude Children's Research Hospital combines technology with teamwork to facilitate studies that advance our understanding of fundamental biological processes and our ability to treat or prevent catastrophic diseases of childhood. The CTIC is a centralized, highly specialized shared resource for the science of light microscopy. The staff consists of the Director of Light Microscopy and three Imaging Scientists. The CTIC has recently undergone a $2.5M renovation, demonstrating a long-term institutional commitment to provide state-of-the-art imaging instrumentation and facilities to the research community at St. Jude Children's Research Hospital. The CTIC is dedicated to helping to solve challenging biological questions with the optimal acquisition and analysis methodologies.

The Director of the Light Microscopy Facility is responsible for managing the light microscopy facility within the institution's Cell and Tissue Imaging Center (CTIC) and for collaborating with faculty and staff on research projects. This center includes equipment for confocal laser scanning microscopy, multiphoton microscopy, spinning disc confocal microscopy, spectral imaging, fluorescence correlation spectroscopy, TIRF, widefield and cell microinjection. Of particular interest is instrumentation optimized for single molecule techniques, such as PALM, STORM and Fluorescence Correlation Spectroscopy. The Director of the Light Microscopy Facility coordinates experimental design for advanced light microscopy; trains faculty and staff on experimental design, light microscopy and data processing and analysis; and maintains image archives.

Requirements:

Bachelor's degree in an appropriate scientific field plus 10 years of post-degree relevant and productive work experience.
OR
Master's degree in an appropriate scientific field plus 9 years of post-degree relevant and productive work experience.
OR
PhD degree in an appropriate scientific field plus 5 years of post-degree relevant and productive work experience.

Preferred skills: Demonstrated excellence in the management of a light microscopy shared resource is preferred. Demonstrated progression in continuing education in advanced light microscopy techniques is preferred.

To find out more this position and to apply, please visit our website, www.stjude.org/jobs.

St. Jude offers a positive working culture, professional advancement and competitive compensation.

St. Jude is an Equal Opportunity Employer and a Drug-Free Workplace.

Candidates receiving offers of employment will be subject to pre-employment drug testing and background checks.


Marta Jarquin-Pardo,PhD


Email Disclaimer: www.stjude.org/emaildisclaimer



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From: David.Llewellyn-at-anu.edu.au
Date: Tue, 30 Nov 2010 22:36:38 -0600
Subject: [Microscopy] 2000ex/fx

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Have to give away complete goniometer stage for 2000 EX/FX all in one
piece, (assembled) contact me direct if you want it, cheers David.

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From: dcd-at-unimelb.edu.au
Date: Wed, 1 Dec 2010 07:31:15 -0600
Subject: [Microscopy] viaWWW: Post-staining with triple lead

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Email: dcd-at-unimelb.edu.au
Name: Chaitali

Organization: Melbourne university

Title-Subject: [Filtered] Post-staining with triple lead

Message: Hello All

Thanks for the reply.

I have tried all different ways to avoid lead precipitation such as
preparing the solution in boiled Milli Q water , spinning down the
lead citrate solution before using it.

However, i was interested in the protocol that suggested to wash
first in 0.01N NaOH after lead staining and then successive washes in
boiled Milli Q water.
My question is wont the NaOH wash precipitate the uranyl acetate
stain? As i read few protocols that stated the Uranyl acetate gets
precipitated in the presence of NaOH?

Thanks

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From: PhillipsT-at-missouri.edu
Date: Wed, 1 Dec 2010 08:00:41 -0600
Subject: [Microscopy] viaWWW: Post-staining with triple lead

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A 0.1 NaOH may be helpful but since it can form NaHCO3 by combining with atmospheric CO2 it may exacerbate the problem if you aren't careful since this can lead to PbCO3 precipitates forming. But it won't impact the uranyl acetate since the original lead solution was equally basic and that doesn't cause problems. I don't believe most people find a NaOH wash necessary so it is probably better to prevent the problem rather than trying to reverse it if possible. Like much of what we do in science, half the steps are unnecessary but we just don't know which half. A NaOH wash might be essential if you have a different less desirable step upstream from this one but unnecessary if you have a working streamlined approach. Be careful that you aren't breathing on your grids while staining them since the CO2 in your breath causes precipitates. I have caught many students intently staring at the grids from a short distance while trying to pick them up off a lead citrate drop. Everyone who has ever done a lot of TEM has had this problem. There are dozens of papers on how to prevent it which tells you that there is no one method that works for all or we wouldn't need to keep coming up with the solution. Good luck. Tom Phillips




Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


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Email: dcd-at-unimelb.edu.au
Name: Chaitali

Organization: Melbourne university

Title-Subject: [Filtered] Post-staining with triple lead

Message: Hello All

Thanks for the reply.

I have tried all different ways to avoid lead precipitation such as
preparing the solution in boiled Milli Q water , spinning down the
lead citrate solution before using it.

However, i was interested in the protocol that suggested to wash
first in 0.01N NaOH after lead staining and then successive washes in
boiled Milli Q water.
My question is wont the NaOH wash precipitate the uranyl acetate
stain? As i read few protocols that stated the Uranyl acetate gets
precipitated in the presence of NaOH?

Thanks

Login Host: 60.228.112.27
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From: david.wilbur-at-tufts.edu
Date: Wed, 1 Dec 2010 08:29:07 -0600
Subject: [Microscopy] SEM/EDS Need port cover for JEOL 840 microscope

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Fellow EM listers:

My Oxford EDS x-ray detector needs to be returned to Oxford for repair,
and unfortunately, I cannot locate the original cover for the EDS port
on the microscope. When this happened eight years ago, neither Oxford
nor JEOL could supply a cover, but I was able to obtain a loaner through
the generosity of this group. I do not want to lose the use of the SEM
for imaging while the detector is repaired. The microscope is a JEOL
JSM-840A. Is there anyone out there able and willing to help me with a
sale, rent, or loan of a suitable cover until my x-ray detector is repaired.

Thank you.

Dave Wilbur

--
__________________________________
David J. Wilbur, Ph.D.
Instrumentation Specialist
Department of Chemistry
Tufts University
voice: 617-627-2163
Fax: 617-627-3443
email: dwilbu01-at-tufts.edu
__________________________________



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From: oshel1pe-at-cmich.edu
Date: Wed, 1 Dec 2010 08:43:58 -0600
Subject: [Microscopy] Re: SEM/EDS Need port cover for JEOL 840 microscope

Contents Retrieved from Microscopy Listserver Archives
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Dave,

I have one. Let me know if you don't find a loaner that's more handy
than where I am in the middle of the mitten.

Phil

} Fellow EM listers:
}
} My Oxford EDS x-ray detector needs to be returned to Oxford for repair,
} and unfortunately, I cannot locate the original cover for the EDS port
} on the microscope. When this happened eight years ago, neither Oxford
} nor JEOL could supply a cover, but I was able to obtain a loaner through
} the generosity of this group. I do not want to lose the use of the SEM
} for imaging while the detector is repaired. The microscope is a JEOL
} JSM-840A. Is there anyone out there able and willing to help me with a
} sale, rent, or loan of a suitable cover until my x-ray detector is repaired.
}
} Thank you.
}
} Dave Wilbur
}
} --
} __________________________________
} David J. Wilbur, Ph.D.
} Instrumentation Specialist
} Department of Chemistry
} Tufts University
} voice: 617-627-2163
} Fax: 617-627-3443
} email: dwilbu01-at-tufts.edu
} __________________________________

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: ryan.deacon-at-jhuapl.edu
Date: Wed, 1 Dec 2010 15:40:55 -0600
Subject: [Microscopy] viaWWW: e-beam lithography systems

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Email: ryan.deacon-at-jhuapl.edu
Name: Ryan Deacon

Organization: Johns Hopkins University Applied Physics Lab

Title-Subject: [Filtered] e-beam lithography systems

Message: Hello List Members -

We will be purchasing an e-beam lithography system for our SEM in
the next few months. I am looking for feedback from any users of the
Nabity and Raith systems to help us with our decision - why did you
chose one over the other, are you still happy with your decision, any
issues regarding service and support, etc. Off line responses are
fine.

The lithography system will be used on a Hitachi S4700, without a
beam blanker. We are aware of the beam stability issues and the
limitations of not having the blanker - these will not pose a problem
for our intended use. And we will eventually be moving the e-beam
system over to a new thermal FEG.

Thanks in advance - Ryan


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From: gary-at-gaugler.com
Date: Wed, 1 Dec 2010 16:55:07 -0600
Subject: [Microscopy] Re: viaWWW: e-beam lithography systems

Contents Retrieved from Microscopy Listserver Archives
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You have a rather complicated situation and perhaps
a dilemma. One problem with the 4700 is poor stage
accuracy. This is only a problem or issue when
doing writing. If you want to move the nabity to
another SEM then the nabity would have to be able
to live with it. Again, stage accuracy and alignment
will still be a challenge. since nabity does not
supply a stage but rather relies on the SEM stage,
and a serial interface to control it.

A better but more costly system is the Raith C-100.
It is based on a Zeiss Supra column and plinth. Raith
supplies (attached) a special chamber to hold the
mask or wafer. The stage is laser aligned and moved
via piezo motors. The combination results in very
high accuracy and the ability to write extremely fine
feature sizes. The Schottky FE and high current option
offer great stability and short write time. Their
high speed beam blanker makes a job pretty much hands-off.

A Raith stage alone is also an option but has to be fit to the
SEM to replace the door and original SEM stage. Whether
or not that stage+door will fit on your new FESEM is
unknown to me. But the Raith stage would solve any
stage issues with the 2700. But depending on what you
want to do, you may still need a beam blanker.

If you get a thermal FE and it has magnetic immersion
final lens, that may be an issue with nabity and with
the Raith stage. That is probably why they use the
Zeiss Supra Gemini column because it is an electrostatic
system. Another nice feature is that it does not
have any final apertures.

gary g.

At 01:42 PM 12/1/2010, you wrote:



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From: germpore-at-sonic.net
Date: Thu, 2 Dec 2010 05:44:45 -0600
Subject: [Microscopy] Substitute for Chloral Hydrate?

Contents Retrieved from Microscopy Listserver Archives
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I'm finding that chloral hydrate is increasingly difficult to purchase
even for a school department, being a Schedule IV Drug. This is
unfortunate, as it was once a basic compound for use in microscopy,
for solutions such as Hoyer's mounting media, Melzer's reagent, or on
its own in an aqueous solution. Does anybody have suggestions for
other compounds that might serve as a good clearing agent, without
depending on being a strong acid or base for their clearing activity?

Thanks,
Peter

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From: DOrloff-at-ascb.org
Date: Thu, 2 Dec 2010 08:15:09 -0600
Subject: [Microscopy] Microscopy Image Repository

Contents Retrieved from Microscopy Listserver Archives
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With apologies for cross-posting.

Got Images?

Dear Colleague,

Won't you help us build a new valuable resource for researchers, students, and the public?

The Cell: An Image Library(tm) (www.cellimagelibrary.org) is a new research-, reference-, and education-focused cell image, video, and animation repository. An open-access resource for peer-reviewed images and their raw data, The Cell will help researchers share and archive their data. It will create a common file format for use by all. And its growing contents will be easy to search.

With your images-published and unpublished, we can further discovery and education.

The Cell: An Image Library(tm) is now accepting images for annotation and publication. We seek data from all organisms, cell types, microscopy modalities, and disease states. Uploading images is easy and will only take a few minutes. Our annotators peer review each unpublished image, and contributors will be acknowledged on the website. The Cell's contents can be cited in articles; contributions can be noted on CVs.

Won't you help your community?

Visit http://www.cellimagelibrary.org/pages/contribute for instructions and the link to submit your image data. Please see http://www.cellimagelibrary.org/pages/license for information about licensing options. Raw data as well as data with limited processing will be the most valuable. We accept images from 105 different formats. Thank you in advance for your contributions. If you have any questions or suggestions, please email ASCB Manager, Image Library, David Orloff at dorloff-at-ascb.org.

Sincerely,

Timothy J. Mitchison Caroline Kane Joan R. Goldberg
President, ASCB PI, The Cell Executive Director, ASCB

The Cell is funded by NIGMS Grand Opportunities grant RC2GM092708 to the American Society for Cell Biology (ASCB).



David Orloff
Manager, Image Library
The American Society for Cell Biology
8120 Woodmont Avenue, Suite 750
Bethesda, MD 20814-2762
T: 301-347-9300/Direct: 301-347-9305
F: 301-347-9310
E-mail: dorloff-at-ascb.org
Web site: www.ascb.org
The Cell: An Image Library(tm): www.cellimagelibrary.org

Don't miss the 50th ASCB Annual Meeting in Philadelphia, Dec. 11-15, 2010!



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From: oshel1pe-at-cmich.edu
Date: Thu, 2 Dec 2010 08:45:09 -0600
Subject: [Microscopy] Re: Substitute for Chloral Hydrate?

Contents Retrieved from Microscopy Listserver Archives
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Peter,

We use methyl salicylate. Around here, mostly for embryos and shrimp.
Works well for fluorescence microscopy, too.
It does require dehydration with ethanol.

Phil

} I'm finding that chloral hydrate is increasingly difficult to purchase
} even for a school department, being a Schedule IV Drug. This is
} unfortunate, as it was once a basic compound for use in microscopy,
} for solutions such as Hoyer's mounting media, Melzer's reagent, or on
} its own in an aqueous solution. Does anybody have suggestions for
} other compounds that might serve as a good clearing agent, without
} depending on being a strong acid or base for their clearing activity?
}
} Thanks,
} Peter

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: cvierret-at-mst.edu
Date: Thu, 2 Dec 2010 09:11:11 -0600
Subject: [Microscopy] viaWWW: Au/Pd Targets

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Name: Clarissa Wisner

Organization: MS&T

Title-Subject: [Filtered] Au/Pd Targets

Message: What does you lab do with worn out Au/Pd targets?

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From: oshel1pe-at-cmich.edu
Date: Thu, 2 Dec 2010 09:50:43 -0600
Subject: [Microscopy] Re: viaWWW: Au/Pd Targets

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Either sell them for the Au and Pd content, or send them to the
vendor from whom we're buying a new target to get money off the price
of the new target.

Phil

} This Question/Comment was submitted to the Microscopy Listserver
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} Email: cvierret-at-mst.edu
} Name: Clarissa Wisner
}
} Organization: MS&T
}
} Title-Subject: [Filtered] Au/Pd Targets
}
} Message: What does you lab do with worn out Au/Pd targets?
}

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: per.horstedt-at-medbio.umu.se
Date: Thu, 2 Dec 2010 10:11:19 -0600
Subject: [Microscopy] broken tapestreamer

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi all,
I need to replace the tape-streamer in an older Link AN10000 EDX system.
The original streamer is a Tandberg TDC 3610.
If someone has a working one to spare (or an equivalent) or can tell me
where to get one, please contact me off-list.

Cheers from a wintry northern Sweden,
Per Hörstedt

EM Platform
KBC Building
University of Umea
S-90186 Umea
Sweden

phone: int-46-90-786 5670
mail: per.horstedt-at-medbio.umu.se



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From: germpore-at-sonic.net
Date: Thu, 2 Dec 2010 14:13:24 -0600
Subject: [Microscopy] Thiodiethanol question

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

So I inquired the other day about chloral hydrate substitutes, and one
of the compounds that sounds promising is 2,2-thiodiethanol (aka TED
or thiodiglycol). (A big thanks to Keith Duncan for the tip, BTW.)
Article here:

http://onlinelibrary.wiley.com/doi/10.1002/jemt.20396/pdf

The question I have about it is, does anybody know whether this
compound is reactive with iodine or potassium iodide if they were
mixed in the same aqueous solution? I'm not a chemist, so I wouldn't
know offhand from looking at the classes of compound whether the two
are reactive. My concern is that they are miscible without reacting
and either creating another compound or creating a cloudy precipitate.
Or worse, creating a compound that's highly toxic. (I will note that
TDE is a precursor of mustard gas, which is why I'm particularly
concerned that it not be reactive with a halogen like iodine.)

Has anybody used TDE with iodine stains?

Peter

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From: edelmare-at-muohio.edu
Date: Thu, 2 Dec 2010 14:16:49 -0600
Subject: [Microscopy] TEM Sample Temperature

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html


O.k., I have a user who is trying to compare TEM images that he and I took
with those he has found in the literature. And he asks "I was trying to
compare my data from what I found in the literature and it seems to depend
on temperature (20C vs 30C vs 40C). Do you by chance know about what
temperature the images were taken at?"

So please, educate me. To my knowledge accurately holding and knowing
the precise temperature (+/- 10C) of the sample while being imaged in a
TEM is unrealistic.

("Knowing" the temperature in most cases, because watching melting points
or boiling points or some other structural change point might provide
accurate sample temperature in special cases.)

There is thermal energy imparted to the samples by the electron beam -
when the electron beam hits them, there are inelastic events between the
atoms of the sample and the primary electrons in the beam. How much
energy per unit time is determined by how much interaction there is,
dependant on the sample, the accelerating voltage (and beam current).
High density (atomic or structural) and/or thicker samples have higher
interaction thus more energy transfer. Low density samples have less.
Then there is then rate of thermal conductivity within the sample. Is one
area being overheated or is it rapidly being dissipated? As has been
discussed numerous times on this Listserv heating can easily be as high as
+1800C (the melting point of glass).

And cryo stage and thermal stages do not ensure the samples stay at a
specific temperature while being imaged. There two competing forces - the
thermal stage trying to hold sample temperature and the beam adding heat
to the sample. You have a thermal regulation source (cooling or heating) in
a sample holder which probably does a very good job of regulating the
temperature of the *grid* holding the sample. But then there is a matter of
thermal transfer from the grid to the sample. A watery matrix supporting a
cryo-sample has one thermal conductivity (poor conductivity of ice?) vs
particles supported on a carbon or polymer film (almost no thermal
conductivity?) vs gold crystals in direct contact with the supporting grid
(pretty good conductivity).

So is the statement "I was trying to compare my data from what I found in
literature and it seems to depend on temperature (20C vs 30C vs 40C)." a
little unrealistic? Knowing the sample temperature, in the high vacuum, while
being imaged with the beam? Even if you are using some sort of "low dose"
imaging methodology?

In this case specifically, the samples being imaged are organic bicelles and
being imaged at 20C or 30C or 40C means they were not imaged with a
cryo-stage. "Bicelles are a novel form of long-chain/short-chain phospholipid
aggregates, which are useful for biophysical and biochemical studies of
membrane-associated biomolecules."


Richard E. Edelmann, Ph.D.
Electron Microscopy Facility Director
364 Pearson Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-muohio.edu
http://www.emf.muohio.edu



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From: frah0010-at-umn.edu
Date: Thu, 2 Dec 2010 14:40:07 -0600
Subject: [Microscopy] digital replacement for thermal printers

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Microscopists,

We have an old Sony video thermal printer (UP-870MD) hooked up to our microprobe to make quick prints of the electron image that can be taped into lab notebooks, but it seems to have finally died -- I'll take a look at the power supply and such, but this seems like an opportune time to consider alternatives.

Does anyone know of a digital replacement for these thermal printers? I'm envisioning a replacement that, instead of printing photos on thermal paper, saves the image to a SD card or USB stick. Does such a device exist?

Best,
Ellery

Ellery Frahm, Ph.D.
Research Associate, Department of Geology & Geophysics
Manager & Principal Analyst, Electron Microprobe Laboratory
University of Minnesota - Twin Cities campus
http://probelab.geo.umn.edu/



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From: mike.bode-at-resaltatech.com
Date: Thu, 2 Dec 2010 15:04:46 -0600
Subject: [Microscopy] digital replacement for thermal printers

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html


Ellery,

Do you know the video specification for the Sony? If it is NTSC (or
perhaps PAL), an inexpensive PC card would be able to capture the image
and save to a file. If not NTSC (eg. slow scan), it might take a more
sophisticated (translate costly) card, but still could be done.

Woody


N.W. (Woody) White Jr.
Senior Electron Microscopist
Chemistry and Materials Center
AREVA NP Inc
An AREVA and Siemens Company
Lynchburg, VA
Lab Phone: 434.832.3004

-----Original Message-----
X-from: frah0010-at-umn.edu [mailto:frah0010-at-umn.edu]
Sent: Thursday, December 02, 2010 3:46 PM
To: WHITE Norvell (RS/IB)

Microscopists,

We have an old Sony video thermal printer (UP-870MD) hooked up to our
microprobe to make quick prints of the electron image that can be taped
into lab notebooks, but it seems to have finally died -- I'll take a
look at the power supply and such, but this seems like an opportune time
to consider alternatives.

Does anyone know of a digital replacement for these thermal printers?
I'm envisioning a replacement that, instead of printing photos on
thermal paper, saves the image to a SD card or USB stick. Does such a
device exist?

Best,
Ellery

Ellery Frahm, Ph.D.
Research Associate, Department of Geology & Geophysics Manager &
Principal Analyst, Electron Microprobe Laboratory University of
Minnesota - Twin Cities campus http://probelab.geo.umn.edu/



==============================Original
Headers==============================
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{frah0010-at-umn.edu} 7, 19 -- Content-Type: text/plain; charset=us-ascii
7, 19 -- Subject: digital replacement for thermal printers 7, 19 --

Hello Ellery,

I think you have 3 options:

1) You could replace the printer with the same or similar printer. I just
googled it and there seems to be a used one for sale here:
http://www.dotmed.com/listing/539198 (No affiliation, just saw that on the
web. Contact at your own risk).

2) I assume that the printer connects to a standard TV signal output and
prints from there. If that is the case, and you want to grab the image and
store it digitally, you would have to buy a "frame grabber" and use software
to read the data from the grabber into a computer and store it from there.
This is fairly cheap for a regular TV signal.

3) To acquire higher resolution/better SNR images from your probe, you would
probably have to use a dedicated SEM (or probe) interface, such as our
ADDA3. These interfaces, however, are more expensive than the first two
options.

If you have any questions, feel free to give me a call.

Mike
---
Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-748-4346
f: +1-303-202-6350
Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com





-----Original Message-----
X-from: frah0010-at-umn.edu [mailto:frah0010-at-umn.edu]
Sent: Thursday, December 02, 2010 1:47 PM
To: mike.bode-at-resaltatech.com

Microscopists,

We have an old Sony video thermal printer (UP-870MD) hooked up to our
microprobe to make quick prints of the electron image that can be taped into
lab notebooks, but it seems to have finally died -- I'll take a look at the
power supply and such, but this seems like an opportune time to consider
alternatives.

Does anyone know of a digital replacement for these thermal printers? I'm
envisioning a replacement that, instead of printing photos on thermal paper,
saves the image to a SD card or USB stick. Does such a device exist?

Best,
Ellery

Ellery Frahm, Ph.D.
Research Associate, Department of Geology & Geophysics
Manager & Principal Analyst, Electron Microprobe Laboratory
University of Minnesota - Twin Cities campus
http://probelab.geo.umn.edu/



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From: William.F.Tivol-at-aero.org
Date: 12/02/2010 12:28 PM
Subject: [Microscopy] Thiodiethanol question

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Peter,
As also not a chemist, I cannot translate between the names and
the chemical structure of TDE; however, if it is the sulfur equivalent of
ethylene glycol--that is, with two SH groups on it--it should be a
powerful reducing agent, which would react strongly with an oxidizing
agent like iodine.
Yours,
Bill



X-from: germpore-at-sonic.net
To: William.F.Tivol-at-aero.org






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So I inquired the other day about chloral hydrate substitutes, and one
of the compounds that sounds promising is 2,2-thiodiethanol (aka TED
or thiodiglycol). (A big thanks to Keith Duncan for the tip, BTW.)
Article here:

http://onlinelibrary.wiley.com/doi/10.1002/jemt.20396/pdf

The question I have about it is, does anybody know whether this
compound is reactive with iodine or potassium iodide if they were
mixed in the same aqueous solution? I'm not a chemist, so I wouldn't
know offhand from looking at the classes of compound whether the two
are reactive. My concern is that they are miscible without reacting
and either creating another compound or creating a cloudy precipitate.
Or worse, creating a compound that's highly toxic. (I will note that
TDE is a precursor of mustard gas, which is why I'm particularly
concerned that it not be reactive with a halogen like iodine.)

Has anybody used TDE with iodine stains?

Peter

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From: rcommon-at-msu.edu
Date: Thu, 2 Dec 2010 15:25:34 -0600
Subject: [Microscopy] Re: Thiodiethanol question

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I don't have an answer to your specific question, but I have extensive
experience using a variety of iodine reagents for working with fungi and
lichens (see Mycotaxon 42: 35-41. COMMON, R. S. 1991. ) Different
iodine reagents give different results in many cases. The reactions are
very sensitive to the concentration of iodine and the other components
of the reagent. I discuss a variety of reagents in the paper. For most
of my purposes Melzer's reagent was not the most useful, and it is
certainly not the best for detecting many reactive materials. In any
case I doubt you could find another combination of components that would
give precisely the same results as Melzer's.

Ralph Common

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From: frah0010-at-umn.edu
Date: Thu, 2 Dec 2010 15:28:56 -0600
Subject: [Microscopy] RE: digital replacement for thermal printers

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Thanks for the replies so far.

To clarify, it is, as Woody noted, a "slow scan" printer that allows for higher resolution than TV scan rates like NTSC or PAL. A few years ago, I tried a simple video capture card that captured the "TV rate" scan from the microprobe, and the result was, well, quite disappointing. The systems that directly tap into the signal (e.g., Orion) are too expensive and really are overkill for the capability we need to replace -- a quick and easy way to grab an image for one's notes. So I'd like to still take advantage of the higher resolution that this printer is (well, was) capable of printing. I've found that used models are available as well as new models, but I didn't want to overlook any digital options.

Best,
Ellery


Ellery Frahm, Ph.D.
Research Associate, Department of Geology & Geophysics
Manager & Principal Analyst, Electron Microprobe Laboratory
University of Minnesota - Twin Cities campus
http://probelab.geo.umn.edu/









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From: William.F.Tivol-at-aero.org
Date: 12/02/2010 12:28 PM
Subject: [Microscopy] TEM Sample Temperature

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Dear Richard,
I do not have my reference books with the relevant information
with me, but calculating the energy imparted to the specimen is fairly
straightforward. You can assume that any specimen will absorb the same
amount of energy as another if it is of the same thickness, measured in
units of gram/cm^2. This is not exact, but it will not be a significant
source of error. The datum you need is the energy loss in units of
eV*cm^2/gram for electrons of the appropriate energy. This is listed in
tables as the "stopping power". Some of that energy will go to the
production of secondary electrons, which will leave the specimen, and
other parts of that energy will go to excitations and displacements of
atoms, which will degrade to heat or will produce photons, which will also
leave the specimen. Therefore, if you consider only the stopping power,
and use that to calculate the energy deposited in the specimen, you will
get an upper limit to the heat introduced by the beam. The heat lost will
be due to conduction, convection, and radiation, of which the first
process will be the largest. For low dose cryo imaging, the conduction is
limited by the poor conductivity of ice, as you said, but even so, a
specimen embedded in amorphous ice and maintained at ~-180 C will not be
heated enough to convert the ice to the hexagonal crystal form, which
occurs at ~-130 C (if I remember correctly). A small condensed beam will
sublimate the ice where it is incident on the specimen, but will not cause
the amorphous-crystalline transition in the surrounding ice, so a high
local dose will deposit a lot of energy, but that energy is not
efficiently conducted as heat to nearby areas. One can certainly image
using a low dose rate, i.e., low beam current, without increasing the
temperature by as much as 50 degrees, and I am confident that with a
sufficiently low dose, one can make the temperature increase smaller than
10 degrees, but, as I said, I do not at present have access to the
stopping power tables to calculate this.
Yours,
Bill



X-from: edelmare-at-muohio.edu
To: William.F.Tivol-at-aero.org




O.k., I have a user who is trying to compare TEM images that he and I took

with those he has found in the literature. And he asks "I was trying to
compare my data from what I found in the literature and it seems to depend

on temperature (20C vs 30C vs 40C). Do you by chance know about what
temperature the images were taken at?"

So please, educate me. To my knowledge accurately holding and knowing
the precise temperature (+/- 10C) of the sample while being imaged in a
TEM is unrealistic.

("Knowing" the temperature in most cases, because watching melting points

or boiling points or some other structural change point might provide
accurate sample temperature in special cases.)

There is thermal energy imparted to the samples by the electron beam -
when the electron beam hits them, there are inelastic events between the
atoms of the sample and the primary electrons in the beam. How much
energy per unit time is determined by how much interaction there is,
dependant on the sample, the accelerating voltage (and beam current).
High density (atomic or structural) and/or thicker samples have higher
interaction thus more energy transfer. Low density samples have less.
Then there is then rate of thermal conductivity within the sample. Is one

area being overheated or is it rapidly being dissipated? As has been
discussed numerous times on this Listserv heating can easily be as high as

+1800C (the melting point of glass).

And cryo stage and thermal stages do not ensure the samples stay at a
specific temperature while being imaged. There two competing forces - the

thermal stage trying to hold sample temperature and the beam adding heat
to the sample. You have a thermal regulation source (cooling or heating)
in
a sample holder which probably does a very good job of regulating the
temperature of the *grid* holding the sample. But then there is a matter
of
thermal transfer from the grid to the sample. A watery matrix supporting
a
cryo-sample has one thermal conductivity (poor conductivity of ice?) vs
particles supported on a carbon or polymer film (almost no thermal
conductivity?) vs gold crystals in direct contact with the supporting grid

(pretty good conductivity).

So is the statement "I was trying to compare my data from what I found in
literature and it seems to depend on temperature (20C vs 30C vs 40C)." a
little unrealistic? Knowing the sample temperature, in the high vacuum,
while
being imaged with the beam? Even if you are using some sort of "low dose"

imaging methodology?

In this case specifically, the samples being imaged are organic bicelles
and
being imaged at 20C or 30C or 40C means they were not imaged with a
cryo-stage. "Bicelles are a novel form of long-chain/short-chain
phospholipid
aggregates, which are useful for biophysical and biochemical studies of
membrane-associated biomolecules."


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From: beth-at-plantbio.uga.edu
Date: Fri, 3 Dec 2010 10:35:33 -0600
Subject: [Microscopy] Fuji film for SEM

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Hi all,
Has anyone used the peel apart Fuji film for SEM? If yes, are you
using the FP-100B or some other?
thanks in advance for all replies,
Beth

Beth Richardson
Plant Biology EM Lab
University of Georgia
Athens, GA 30602


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From: pmcooke-at-earthlink.net
Date: Fri, 3 Dec 2010 11:58:39 -0600
Subject: [Microscopy] Video camera repair service?

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Anyone know where I can get a Javelin "SmartCam", model # JE3762DSP,
serviced? Looks like someone plugged it into a power source that was
defective. I use it for projecting images from light microscopes.

Thank you,

Peter
Peter M. Cooke
MICA
pmcooke-at-earthlink.net



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5, 28 -- Subject: Video camera repair service?
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From: kenconverse-at-qualityimages.biz
Date: Fri, 3 Dec 2010 12:56:08 -0600
Subject: [Microscopy] Video camera repair service?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Peter,
Look at http://www.photomachining.com/PMI2000F.html
It may be cheaper than repair.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: pmcooke-at-earthlink.net [mailto:pmcooke-at-earthlink.net]
Sent: Friday, December 03, 2010 1:01 PM
To: kenconverse-at-qualityimages.biz

Anyone know where I can get a Javelin "SmartCam", model # JE3762DSP,
serviced? Looks like someone plugged it into a power source that was
defective. I use it for projecting images from light microscopes.

Thank you,

Peter
Peter M. Cooke
MICA
pmcooke-at-earthlink.net



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From: ehaller-at-health.usf.edu
Date: Fri, 3 Dec 2010 15:15:03 -0600
Subject: [Microscopy] Fuji film for SEM

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi, Beth,

We've been using the film for a few months now. Fuji just discontinued making the FP100B black and white film in 4x5 format, due to poor sales. You can still purchase their 4x5 color film. The black and white version gave us similar results to Polaroid Type 55 film, but with a somewhat reduced palette of grey tones. We were satisfied with the results. Resolution was fine. We are switching to the color film for our old SEM, and will use an orange filter to render our images true black and white, since the phosphor on our CRT has a bluish tinge to it. I know that if we scan our images in Photoshop, the tinge is of no consequence. We had to purchase a film back for the film. I don't know if the old Polaroid 505 film back will hold the new Fuji film. I don't have one of these older backs.

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: LSA 119
________________________________________
X-from: beth-at-plantbio.uga.edu [beth-at-plantbio.uga.edu]
Sent: Friday, December 03, 2010 11:45 AM
To: Haller, Edward

Hi all,
Has anyone used the peel apart Fuji film for SEM? If yes, are you
using the FP-100B or some other?
thanks in advance for all replies,
Beth

Beth Richardson
Plant Biology EM Lab
University of Georgia
Athens, GA 30602


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From: maloneyb-at-fiu.edu
Date: Mon, 6 Dec 2010 15:03:10 -0600
Subject: [Microscopy] Microscope with camera lucida for sale

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Group: does anyone have a microscope for sale that has an attached
camera lucida?
Thanks so much.
Barbara


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From: schooley-at-mcn.org
Date: Mon, 6 Dec 2010 15:39:11 -0600
Subject: [Microscopy] Microscopy: Education

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Have you just realized that you've forgotten to get a holiday present
for your favorite grandchild? Don't panic - just visit Project
MICRO's booklists ('Microscopy for Children', 'Nanotechnology for
Kids') at http://www.microscopy.org/ProjectMICRO . Many internet
book dealers have fast shipping. If you need specific advice, I
promise to answer your Email promptly!
--
Caroline Schooley
Project MICRO Coordinator
Microscopy Society of America
45301 Caspar Point Road, Box 117
Caspar, CA 95420
Project MICRO: http://www.microscopy.org/ProjectMICRO

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From: marksmsa-at-gmail.com
Date: Mon, 6 Dec 2010 17:50:12 -0600
Subject: [Microscopy] Postdoctoral Position: Have Sample, Will Travel

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Postdoctoral Position: Have Sample, Will Travel

A slightly different type of postdoctoral position “Have Sample, Will
Travel” is available starting in 2011 at Northwestern University joint
with Argonne National Labs.

The idea of the position is to work jointly with the group of L. D.
Marks (www.numis.northwestern.edu), and various catalytic chemists in
the Institute for Atom-Efficient Chemical Transformations
(http://www.anl.gov/catalysis-science/) on a range of different
materials where we are trying to deliberately control the size, shape
and distribution of catalysts at the atomic scale by a variety of
different techniques. The job of the postdoctoral scientist will be to
interact with the groups synthesizing the systems to produce samples
appropriate for study by electron microscopy, find the appropriate
instrument to obtain the required information (e.g. Cs-corrected STEM,
HREM, EFTEM, EELS etc) then interact with scientists elsewhere to
collect the appropriate data, for instance by travelling to different
DOE laboratories or other Universities where specialized instruments
are available.

Requirements for the position include:
a) Strong communication abilities, since the scientist will need to
interact with many scientists who are not TEM specialists.
b) A strong background in state-of-the-art electron microscopy techniques.
c) A strongly motivated, self starter.

Additional background in heterogeneous catalysis is useful, but not essential.

Applicants should send a CV by email to L-marks-at-northwestern.edu which
should include their three best papers as well as the names and emails
of three referees.


--
Professor Laurence Marks
Department of Materials Science and Engineering
MSE Rm 2036 Cook Hall
2220 N Campus Drive
Northwestern University
Evanston, IL 60208, USA
Tel: (847) 491-3996 Fax: (847) 491-7820
email: L-marks at northwestern dot edu
Web: www.numis.northwestern.edu
EMM2007 http://ns.crys.ras.ru/EMMM07/
Commission on Electron Diffraction of IUCR
www.numis.northwestern.edu/IUCR_CED


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9, 33 -- Subject: Postdoctoral Position: Have Sample, Will Travel
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From: cvierret-at-mst.edu
Date: Mon, 6 Dec 2010 19:18:08 -0600
Subject: [Microscopy] viaWWW: EDAX and S4700

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Email: cvierret-at-mst.edu
Name: Clarissa Wisner

Organization: MS&T

Title-Subject: [Filtered] EDAX and S4700

Message: We are getting a new objective lens today. It should help
with our stigmation problem. My question is do others have a SDD
EDAX system on a S4700 and if so what is your count rate and how do
you configure the scope for that count rate.

Thanks,

Clarissa Wisner

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From: ralf.theissmann-at-uni-due.de
Date: Mon, 6 Dec 2010 19:18:12 -0600
Subject: [Microscopy] viaWWW: EDX Quantification

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Email: ralf.theissmann-at-uni-due.de
Name: Ralf Theissmann

Organization: University of Duisburg-Essen

Title-Subject: [Filtered] EDX Quantification

Message: Dear Listers,
I have a sample containing Ge and Au. The overlap of EDX peaks makes
it reasonable use the Gold M-Line and Ge-L line for qunatification.
Unfotunately, ES-Vision doesn't provide a k-factor for the Au M-line.
Can anyone provide a k-factor or is it reasonable to use one of a
line close to it?
Thanks,
Ralf

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From: gary-at-gaugler.com
Date: Mon, 6 Dec 2010 19:41:43 -0600
Subject: [Microscopy] Re: viaWWW: EDAX and S4700

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Which SDD detector and which DPP?

For SEMs like the 4700, the control options are
final condenser lens current (spot size) and final
aperture size.

gary g.


At 05:19 PM 12/6/2010, you wrote:



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From: swalck-at-southbaytech.com
Date: Mon, 6 Dec 2010 20:31:23 -0600
Subject: [Microscopy] Ionization gauge question...

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One of the controllers is set wrong. You set the emission for the ion
gauge, not the controller. The value of 10 is a common setting and I bet
that is the correct value. You need to look up the value for the gauge.

-Scott

Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA 92673

eMail: SWalck-at-SouthBayTech.com
Phone: 949-492-2600
Toll Free: 1-800-728-2233 (USA)
Fax: 949-492-1499

www.SouthBayTech.com



-----Original Message-----
X-from: jflaci-at-ms.sapientia.ro [mailto:jflaci-at-ms.sapientia.ro]
Sent: Wednesday, August 04, 2010 2:19 AM
To: swalck-at-southbaytech.com

Hi everybody!

We have in our lab an ULVAC ionization gauge controller, GI-N3.
We recently aquired another vacuummeter, and it shows almost double of the
pressure measured with the ionization gauge.

Until now we have set the emission of the ionization gauge to 5 when
the emission check
button was depressed. But now I wonder if it should set to 10 the same
way, as it reads then
almost the same pressure as the other gauge.

Does anybody have a user manual for this type of controller? If so,
could provide me the
necessary details from it?

Thanks

Jakab-Farkas Laszlo

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From: Artur.irani-at-yahoo.com
Date: Tue, 7 Dec 2010 04:04:26 -0600
Subject: [Microscopy] viaWWW: EPMA

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Name: kazem

Organization: IMPRC

Title-Subject: [Filtered] EPMA

Message: Hi
Is there any one to help me for solving some EPMA technical problems?
i am mineralogist and working with EPMA Sx100.
best regards

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From: nicholas.ritchie-at-nist.gov
Date: Tue, 7 Dec 2010 08:07:40 -0600
Subject: [Microscopy] RE: viaWWW: EDX Quantification

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It sounds like what you are interested in are the ZAF quantitative correction factors. However...
1. The correction factors depend on the composition so it is necessary to estimate the composition, calculate the corrections, then iteratively improve the estimated composition and recalculate the corrections until the composition computed k-ratios match the measured k-ratios. This is how all vendors software does it.
2. You need to specify additional information such as the detector take-off angle and beam energy.

A better way to approach this problem is to use my tool DTSA-II (available from http://www.cstl.nist.gov/div837/837.02/epq/dtsa2/index.html) to perform the quantification assuming you have or can collect standard spectra for both Au and Ge. After you have configured a detector, the "Quantification Alien" will step you though the process. See the documentation page at the above web site for details.

Finally, if you use standards you will probably get more accurate results quantifying using the Au L and Ge K than the Au M because the ZAF corrections are smaller due to reduced absorption. Fitting to high quality standards does a bang up job of resolving even the worst overlaps.

Nicholas Ritchie
National Institute of Standards and Technology


} -----Original Message-----
} From: ralf.theissmann-at-uni-due.de [mailto:ralf.theissmann-at-uni-due.de]
} Sent: Monday, December 06, 2010 8:21 PM
} To: Ritchie, Nicholas
} Subject: [Microscopy] viaWWW: EDX Quantification
}
}
}
}
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} Email: ralf.theissmann-at-uni-due.de
} Name: Ralf Theissmann
}
} Organization: University of Duisburg-Essen
}
} Title-Subject: [Filtered] EDX Quantification
}
} Message: Dear Listers,
} I have a sample containing Ge and Au. The overlap of EDX peaks makes
} it reasonable use the Gold M-Line and Ge-L line for qunatification.
} Unfotunately, ES-Vision doesn't provide a k-factor for the Au M-line.
} Can anyone provide a k-factor or is it reasonable to use one of a
} line close to it?
} Thanks,
} Ralf
}
} Login Host: 134.91.65.66
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} ==============================Original
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5, 29 -- Subject: RE: [Microscopy] viaWWW: EDX Quantification
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From: wesaia-at-iastate.edu
Date: Tue, 7 Dec 2010 08:43:24 -0600
Subject: [Microscopy] viaWWW: EDX Quantification

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I am not familiar with the ES-Vision, so I am a little confused about what you are asking.

You say they don't provide a "k-factor" for the Au M line. I would interpret that to mean that they have not included a standard intensity for Au-M that you can calculate a k-ratio from (k-ratio being the ratio of the elemental intensity in your unknown to the intensity in your standard). That is somewhat surprising. Au is readily available; I would have thought they would have included it and maybe skipped some other elements. We have had standardless capability for Au in our EDS systems for the last 20 years or more.

Perhaps, they have not included a stock reference, but you should be able to collect your own standards. If so, follow the instructions carefully. You must incorporate some reference for beam intensity whether by using a beam current reading or referencing back to another element. We had a user who failed to do that step and he shifted our k-ratios by about 25%. (He really should have asked for help first.)

I would echo Ritchie's comments that good EDS software should have little trouble deconvolving the overlap of a K-line series and an L-line series from two elements. If more elements are involved, things can quickly get more difficult. It is always a good idea to review the peak fit by whatever means they allow. Perhaps they show you the fitted spectrum or maybe they let you examine the residuals. (I had to do that yesterday when I saw a shoulder on the side of the Cu L-line. I suspected Na-K and the residuals quickly revealed that to be the case. They improved greatly upon adding Na to the element mix.)

Warren Straszheim

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Email: ralf.theissmann-at-uni-due.de
Name: Ralf Theissmann

Organization: University of Duisburg-Essen

Title-Subject: [Filtered] EDX Quantification

Message: Dear Listers,
I have a sample containing Ge and Au. The overlap of EDX peaks makes
it reasonable use the Gold M-Line and Ge-L line for qunatification.
Unfotunately, ES-Vision doesn't provide a k-factor for the Au M-line.
Can anyone provide a k-factor or is it reasonable to use one of a
line close to it?
Thanks,
Ralf

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From: wesaia-at-iastate.edu
Date: Tue, 7 Dec 2010 09:11:34 -0600
Subject: [Microscopy] viaWWW: EDAX and S4700

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Like Gary said, your primary controls over count rate (which directly depends on beam current) are your choice of objective aperture and your condenser lens (spot size) setting.

I don't have a S4700 or a SDD detector, so I cannot give you exact settings for particular results. You might also need to tell us which SDD detector you have. Count-rate will depend on the detector size. 10 mm2 used to be standard, but there are now 30 and 80-mm2 detectors which would give proportionally more counts. The count rate will also depend on where you position the detector within the column. Count rate falls off with the square of the distance from the sample. This also presumes you have the sample at the correct analytical height for your detector (i.e., your detector is aimed at your sample). If not, the detector collimator will severely cut back your signal.

I would assume you would like to see several thousand counts per second. More counts means short counting times for the same statistics. Like I said, I don't have an SDD or fast pulse processor. I usually set our system to 3000 cps which leads to about 30% dead time which is about my deadtime limit. Bad things start happening above 30%. On our Hitachi S-2460 with a tungsten gun, I typically use aperture 2 (aperture 4 is our smallest one) and a beam current setting of 195 on a scale of 0-255. I suppose that translates to 0.5 nA. We could probably quadruple that count rate before we ran out of beam. You should be able to do even better if you have a field emission gun.

What you will probably find yourself doing is balancing image resolution with x-ray count rate. If you want sharp images, you will have to live with a lower count rate and vice versa. I told you our starting point, but I mostly am doing EDS. If I need better images, I cut back the current to get the sharpness I need, even if it means choking off my beam so that there are few x-rays produced. I increase the current when I need to collect x-rays again.

Warren Straszheim

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Email: cvierret-at-mst.edu
Name: Clarissa Wisner

Organization: MS&T

Title-Subject: [Filtered] EDAX and S4700

Message: We are getting a new objective lens today. It should help
with our stigmation problem. My question is do others have a SDD
EDAX system on a S4700 and if so what is your count rate and how do
you configure the scope for that count rate.

Thanks,

Clarissa Wisner

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From: NEERAJG-at-clemson.edu
Date: Tue, 7 Dec 2010 11:26:16 -0600
Subject: [Microscopy] Re: Abstract Deadline for the 103rd Annual Meeting of the National

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Colleagues,

The abstract submission deadlines for the Biofouling and Shell Formation (Biomineralization) sessions are soon approaching. The abstract submission deadline is Monday 13th of December, the registration for the meeting is now open! Please see the meeting website for further details http://shellfish.org/103rdAnnualMeeting. Please see the announcement below for information on both sessions.


We are pleased to announce  two special sessions; 1) Biofouling  and 2) Shell Formation (Biomineralization) at the 103rd  Annual Meeting of the National Shellfisheries Association (NSA) in March 27-31 2011 in Baltimore, Maryland. (http://shellfish.org/ )



1) We anticipate the Biofouling session to be broad and multidisciplinary focusing on various aspects of marine and fresh water biofouling including but not limited to imaging. Since imaging is relevant to this community, we welcome abstracts on,

A) Imaging studies of fouling organism highlighting cellular and developmental processes
B) Tracking fouling organisms, development of lab and field assays for evaluating new coatings
B) Being multidisciplinary, we also welcome abstracts from folks who are working on the materials aspect of biofouling by engineering new and improved antifouling coatings and polymers, specially characterization of such materials using various microscopy techniques and or biological assays. 

We also welcome abstracts on other aspects of biofouling such as impact of invasive fouling species etc. I will be chairing the Biofouling session so please feel free to contact me if you have any questions, my contact details are in my signature.
 

2) Similarly we anticipate the Shell Formation (Biomineralization) session to be broad and multidisciplinary as well. We welcome abstracts on various aspects of Biomineralization including but not limited to,

A) Cellular and developmental processes involved in Biomineralization
B) Characterization of shells and or other biomineralized structures using various microscopy techniques
C) Physical, biological and  or chemical aspects of the Biomineralization process
D) Effects of ocean acidification on shell formation and biomineralization

We would also like to invite abstracts from the Bone formation community, folks who are working on the comparative cellular and developmental aspects of bone formation and biomineralization.

If you have any questions about the Shell Formation (Biomineralization) session please contact my colleague 

Dr. Andrew S. Mount
Associate Professor
Okeanos Research Group
Department of Biological Sciences
132 Long Hall
Clemson University
Clemson, SC-29631
USA
Email: mount-at-clemson.edu
Phone: 864-656-3597


Please forward this announcement to your friends and collaborators who are working in these areas. More information on the 103rd Annual Meeting of NSA can be found at http://shellfish.org/  


Best Regards,

Neeraj V. Gohad, Ph.D.
Postdoctoral Fellow
Okeanos Research Group
Department of Biological Sciences
132 Long Hall
Clemson University
Clemson,SC-29634
USA
Email: neerajg-at-clemson.edu  
Phone: 864-656-3597

Website: http://www.clemson.edu/okeanos    












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From: Jessica.Riesterer-at-empa.ch
Date: Tue, 7 Dec 2010 17:57:18 -0600
Subject: [Microscopy] viaWWW: AFM: Solution to drift inside the FIB

Contents Retrieved from Microscopy Listserver Archives
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Email: Jessica.Riesterer-at-empa.ch
Name: Jessica Riesterer

Organization: Empa - Thun, Switzerland

Title-Subject: [Filtered] AFM: Solution to drift inside the FIB

Message: A few months ago I had asked the listserver for any
suggestions on eliminating stage drift for my custom-built AFM that
is placed in situ in our Tescan Lyra FIB. The drift only occurred
when tilted towards the ion beam. I wanted to share that a solution
has been found. I have made a new adapter plate to fit the Tescan
stage with the rotation mechanism removed. The weight of our AFM was
not evenly distributed over the stage when at tilt. The new adapter
distributed the weight much better and has eliminated the drift.

Thank you for the suggestions that I received.

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From: cvierret-at-mst.edu
Date: Tue, 7 Dec 2010 17:57:50 -0600
Subject: [Microscopy] viaWWW: EDAX revisited

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Email: cvierret-at-mst.edu
Name: Clarissa Wisner

Organization: MS&T

Title-Subject: [Filtered] EDAX revisited

Message: Sorry i did not put enough information into my original
question. We have the Apollo detector with the Genesis software.
Current count rate is little better tan with the SiLi detector,
2500cps. This is at 15kV, objective aperture 1, emission current
10uA, condenser 1 at wide open. What I wanted to know was is this all
we can expect as far as count rates? When it was installed the
engineer signed off at ~15,000cps. This was wide open and at 30kV,
not the typical working conditions. We also have an oxford on a FEI
FIB and typical count rates area are 25,000-35,000cps, 15kV,
2.7nA,and on the Evex detector using 4pi I just did analysis at
100,000cps at 15kV. So you can see my confusion with the low count
rates, EDAX told mre the detector can onlt detect what is available.
Will the 4700 be able to produce more counts and if so how to
configure the system?

Thanks for all the previous replies.

Clarissa Wisner
Electron Microscope Specialist
Missouri University of Science and Technology
B-20 McNutt
Rolla, Mo 65409



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From: exploratorium-at-tiscali.it
Date: Tue, 7 Dec 2010 17:58:14 -0600
Subject: [Microscopy] viaWWW: WILD M40A MANUAL NEEDED

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Email: exploratorium-at-tiscali.it
Name: Giovanni De Caro, MD

Organization: Exploratorium Luigi MontalbÚ - Casalciprano - CB - Italia

Title-Subject: [Filtered] WILD M40A MANUAL NEEDED

Message: Hi all! i need teh manual for the WILD
M40 A inverted microsocpe. If any of you has one,
better if in PDF. format, please let me know.
Thanks a lot

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From: colijn.1-at-osu.edu
Date: Tue, 7 Dec 2010 19:04:53 -0600
Subject: [Microscopy] Re: viaWWW: EDAX revisited

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Hi Clarissa,

While you indicate the EMISSION current of your source, you don't say
anything about the BEAM current actually incident on the sample. The
best way to measure this is to use a Faraday cup, but you can get a
reasonable comparison by using the same sample in the your various
scopes with the same beam conditions and comparing the absorbed current
signal (nA) with a picoammeter.

My guesses as to the lack of counts
1. you have a lower beam current than in the other scopes.
2. the detector geometry is different (farther away or smaller
cross-sectional area, i.e. smaller solid angle)
3. Something (e.g. a backscatter detector) is shadowing the SSD. We
have one scope where we have to drop the sample below eucentric to do
EDS because the BS detector occludes the EDS detector.

Good luck,
Henk

At 12/7/2010 6:59 PM, cvierret-at-mst.edu wrote:
}
}
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}
} Email: cvierret-at-mst.edu
} Name: Clarissa Wisner
}
} Organization: MS&T
}
} Title-Subject: [Filtered] EDAX revisited
}
} Message: Sorry i did not put enough information into my original
} question. We have the Apollo detector with the Genesis software.
} Current count rate is little better tan with the SiLi detector,
} 2500cps. This is at 15kV, objective aperture 1, emission current
} 10uA, condenser 1 at wide open. What I wanted to know was is this all
} we can expect as far as count rates? When it was installed the
} engineer signed off at ~15,000cps. This was wide open and at 30kV,
} not the typical working conditions. We also have an oxford on a FEI
} FIB and typical count rates area are 25,000-35,000cps, 15kV,
} 2.7nA,and on the Evex detector using 4pi I just did analysis at
} 100,000cps at 15kV. So you can see my confusion with the low count
} rates, EDAX told mre the detector can onlt detect what is available.
} Will the 4700 be able to produce more counts and if so how to
} configure the system?
}
} Thanks for all the previous replies.
}
} Clarissa Wisner
} Electron Microscope Specialist
} Missouri University of Science and Technology
} B-20 McNutt
} Rolla, Mo 65409
}
}

--
Hendrik O. Colijn
www.ceof.ohio-state.edu
OSU Campus Electron Optics Facility colijn.1-at-osu.edu
040 Fontana Labs (614) 292-0674 (V)
116 W. 19th Ave. (614) 292-7523 (F)
Columbus, OH 43210

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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From: tina-at-pbrc.hawaii.edu
Date: Tue, 7 Dec 2010 19:37:55 -0600
Subject: [Microscopy] Re: viaWWW: EDAX revisited

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

On our S-4800 many of our users set the emission current to 20uA, but
sometimes it does not change it much. Our angle to the detector seems
critical, so you might play with that.

Aloha,
Tina

} Message: Sorry i did not put enough information into my original
} question. We have the Apollo detector with the Genesis software.
} Current count rate is little better tan with the SiLi detector,
} 2500cps. This is at 15kV, objective aperture 1, emission current
} 10uA, condenser 1 at wide open. What I wanted to know was is this all
} we can expect as far as count rates? When it was installed the
} engineer signed off at ~15,000cps. This was wide open and at 30kV,
} not the typical working conditions. We also have an oxford on a FEI
} FIB and typical count rates area are 25,000-35,000cps, 15kV,
} 2.7nA,and on the Evex detector using 4pi I just did analysis at
} 100,000cps at 15kV. So you can see my confusion with the low count
} rates, EDAX told mre the detector can onlt detect what is available.
} Will the 4700 be able to produce more counts and if so how to
} configure the system?
}
} Thanks for all the previous replies.
}
} Clarissa Wisner
} Electron Microscope Specialist
} Missouri University of Science and Technology
} B-20 McNutt
} Rolla, Mo 65409
}
}
}
} Login Host: 131.151.110.252
} ---------------------------------------------------------------------------
}
} ==============================Original Headers==============================
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}

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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From: germpore-at-sonic.net
Date: Tue, 7 Dec 2010 19:49:02 -0600
Subject: [Microscopy] Re: viaWWW: WILD M40A MANUAL NEEDED

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Giovanni De Caro, MD writes:

} Title-Subject: [Filtered] WILD M40A MANUAL NEEDED
}
} Message: Hi all! i need teh manual for the WILD
} M40 A inverted microsocpe. If any of you has one,
} better if in PDF. format, please let me know.
} Thanks a lot

There's a website selling this very thing:

http://www.wild-heerbrugg.com/catalog/product_info.php/products_id/Wild-M40-user-manual-131

I don't know of free source.

Peter




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From: gw265-at-cam.ac.uk
Date: Tue, 7 Dec 2010 22:00:34 -0600
Subject: [Microscopy] Resins & difficult-to-infiltrate algae & animal spores

Contents Retrieved from Microscopy Listserver Archives
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Hi all

does anyone have specific advice for what to do with
difficult-to-infiltrate single cells? I'm looking for recommendations on
resins and on time schedules for infiltration and polymerisation.

I'm working with unicellular eukaryotic algae (2-5 microns, cell wall about
100nm thick) and myxozoan spores (10 microns, spore wall about 500nm
thick). Don't have positive evidence for what the walls are made of, but
it's not obviously mineralised. Both are very susceptible to poor
infiltration and extracted-looking cytoplasm. The literature is
uninformative about how long the good published examples have been
infiltrated.

Does "the longer, the better" apply here? Or will that make the extraction
worse? Or wouldn't it matter on the timescale of a week in resin?
Unfortunately with the EM unit closing for Christmas, and the fact the
myxozoan spores come from an endangered species, I don't have the option of
trying different lengths of time to see what's best. I guess if people were
really adamant that longer is definitely better, I could leave them
infiltrating until the unit opens again on the 5th January...

thanks

Giselle

Dr Giselle Walker
University of Cambridge UK
&
University of Sydney, Australia

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From: david.knecht-at-uconn.edu
Date: Wed, 8 Dec 2010 08:58:50 -0600
Subject: [Microscopy] Fluorescence reflection problem

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I have an LED Illumination system that I have built for my Nikon TI microscopes and have an issue that I don't understand that perhaps someone else has insight on. I was looking at some samples recently and found that I had a huge amount of background signal in the GFP channel that I normally don't see- so much so that I could not see the actual fluorescence from the sample. The fluorescence illuminator is a 470nm peak LED (Thorlabs) using a Chroma 49002 GFP filter set. The transmitted light source is a white LED (Thorlabs). I tracked the problem down to reflected light coming through the system from the transmitted light pathway. It turns out that normally, I have a phase ring in place in the condenser and so the excitation light coming through the sample is absorbed and does not cause a problem. However, I had moved the phase ring out of the path that day, and that led to light going through the condenser, up to the collimation optic in front of the white light LED that I use for transmitted light imaging, reflecting off that optic and then going back down through the dichroic and emission filter flooding the image with green light. I am surprised that so much light could be reflecting off the collimation lens, but I am even more surprised that that light could get through both the dichroic and the emission filter. Any thoughts on why this is happening and the best way to solve it? It is not a huge problem because I can simple keep the phase ring in place, but I want to understand what is happening. Would anti-reflection coatings on the collimation optic fix the problem? Dave

Dr. David Knecht
Department of Molecular and Cell Biology
Co-head Flow Cytometry and Confocal Microscopy Facility
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)




==============================Original Headers==============================
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From: kenikea-at-ornl.gov
Date: Wed, 8 Dec 2010 09:25:05 -0600
Subject: [Microscopy] Access to image plate reader

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

After loaning our Ditabis image plate (IP) system to University of
Wisconsin, I have a visitor that needs access to an IP reader during
the next year. I was wondering if there is a reader within easy
driving distance of Knoxville, TN that he might access. Please reply
directly to me at kenikea-at-ornl.gov. Thank you. ED kenik

Microscopy Group
Materials Science and Technology Division
Oak Ridge National Laboratory
100 Bethel Valley Rd., Bldg. 4515, MS-6064
PO Box 2008
Oak Ridge, TN 37831-6064, USA
Tel: 865 574-5066 Fax: 865 576-5413 e-mail: kenikea-at-ornl.gov


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From: david.knecht-at-uconn.edu
Date: Wed, 8 Dec 2010 10:38:45 -0600
Subject: [Microscopy] Re: Fluorescence reflection problem

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

With the phase ring in place to eliminate the reflection, I tried with and without the exciter and you need the exciter to get normal fluorescence images. Without the exciter the background is too high. The LED profile (M470 at link below) makes it look like there should be no signal to leak through the dichroic and emission filters, but there is. The reflection problem with the exciter in place and no phase ring actually causes worse background than the LED with no exciter and no reflection. Dave

http://www.thorlabs.com/images/TabImages/MxxxL2_spectra_dwg-LRG_dwg.png

On Dec 8, 2010, at 10:43 AM, Aryeh Weiss wrote:

} Dear David,
}
} Are you using the excitation filter of that Chroma set, or are you
} depending on the LED being narrow bandwidth? If you are not filtering
} the LED, the nit can have enough light over 500nm to produce this result.
}
} Best regards,
} --aryeh
}
} On 12/8/10 5:02 PM, david.knecht-at-uconn.edu wrote:
} } ----------------------------------------------------------------------------
} }
} }
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe --
} } http://www.microscopy.com/MicroscopyListserver On-Line Help
} } http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } I have an LED Illumination system that I have built for my Nikon TI
} } microscopes and have an issue that I don't understand that perhaps
} } someone else has insight on. I was looking at some samples recently
} } and found that I had a huge amount of background signal in the GFP
} } channel that I normally don't see- so much so that I could not see
} } the actual fluorescence from the sample. The fluorescence illuminator
} } is a 470nm peak LED (Thorlabs) using a Chroma 49002 GFP filter set.
} } The transmitted light source is a white LED (Thorlabs). I tracked
} } the problem down to reflected light coming through the system from
} } the transmitted light pathway. It turns out that normally, I have a
} } phase ring in place in the condenser and so the excitation light
} } coming through the sample is absorbed and does not cause a problem.
} } However, I had moved the phase ring out of the path that day, and
} } that led to light going through the condenser, up to the collimation
} } optic in front of the white light LED tha! t I use for transmitted
} } light imaging, reflecting off that optic and then going back down
} } through the dichroic and emission filter flooding the image with
} } green light. I am surprised that so much light could be reflecting
} } off the collimation lens, but I am even more surprised that that
} } light could get through both the dichroic and the emission filter.
} } Any thoughts on why this is happening and the best way to solve it?
} } It is not a huge problem because I can simple keep the phase ring in
} } place, but I want to understand what is happening. Would
} } anti-reflection coatings on the collimation optic fix the problem?
} } Dave
} }
} } Dr. David Knecht Department of Molecular and Cell Biology Co-head
} } Flow Cytometry and Confocal Microscopy Facility U-3125 91 N.
} } Eagleville Rd. University of Connecticut Storrs, CT 06269
} } 860-486-2200 860-486-4331 (fax)
} }
} }
} }
} }
}
} --
} Aryeh Weiss
} School of Engineering
} Bar Ilan University
} Ramat Gan 52900 Israel
}
} Ph: 972-3-5317638
} FAX: 972-3-7384051
}

Dr. David Knecht
Department of Molecular and Cell Biology
Co-head Flow Cytometry and Confocal Microscopy Facility
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)




==============================Original Headers==============================
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From: raristau-at-ims.uconn.edu
Date: Wed, 8 Dec 2010 15:44:51 -0600
Subject: [Microscopy] TEM holder

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi All,

Does anyone know of a source for the small jeweled pivot-pins used to hold
specimen cup on double-tilt TEM holders? It seems that heavy-handed users in
our multi-user facility break these jewels every year or so, and I am not
satisfied with the hassle of sending out the holder for repair.

I am sure we are not the only ones to experience this. Does anyone have
hints for on-site repair?

Cheers
Roger

Roger A. Ristau, PhD
Electron Microscopy Specialist
Institute of Materials Science
97 North Eagleville Road
University of Connecticut
Storrs, CT 06269
vox: 860-486-5453
fax: 860-486-4745




==============================Original Headers==============================
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From: rosemary.white-at-csiro.au
Date: Wed, 8 Dec 2010 16:17:30 -0600
Subject: [Microscopy] Fluorescence reflection problem

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Haven't been following this thread, but in the new Zeiss upright compound
fluorescence microscopes, there is so much reflection from the top element
of the condenser (why this problem has arisen now, in the very latest
generation of microscopes is an open question), that you have to remove it
from the light path when doing fluorescence. There are similar reflection
problems in the fluorescence dissectors, but to a much lesser extent.
Perhaps try this - rotating the top element out of the way? It does mean
you then have to merge two images if you want to overlay fluorescence with
DIC, which is often impossible with moving fluorescence objects.

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au



On 9/12/10 3:44 AM, "david.knecht-at-uconn.edu" {david.knecht-at-uconn.edu} wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} With the phase ring in place to eliminate the reflection, I tried with and
} without the exciter and you need the exciter to get normal fluorescence
} images. Without the exciter the background is too high. The LED profile
} (M470 at link below) makes it look like there should be no signal to leak
} through the dichroic and emission filters, but there is. The reflection
} problem with the exciter in place and no phase ring actually causes worse
} background than the LED with no exciter and no reflection. Dave
}
} http://www.thorlabs.com/images/TabImages/MxxxL2_spectra_dwg-LRG_dwg.png
}
} On Dec 8, 2010, at 10:43 AM, Aryeh Weiss wrote:
}
} } Dear David,
} }
} } Are you using the excitation filter of that Chroma set, or are you
} } depending on the LED being narrow bandwidth? If you are not filtering
} } the LED, the nit can have enough light over 500nm to produce this result.
} }
} } Best regards,
} } --aryeh
} }
} } On 12/8/10 5:02 PM, david.knecht-at-uconn.edu wrote:
} } } ----------------------------------------------------------------------------
} } }
} } }
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } } To Subscribe/Unsubscribe --
} } } http://www.microscopy.com/MicroscopyListserver On-Line Help
} } } http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } } ----------------------------------------------------------------------------
} } }
} } } I have an LED Illumination system that I have built for my Nikon TI
} } } microscopes and have an issue that I don't understand that perhaps
} } } someone else has insight on. I was looking at some samples recently
} } } and found that I had a huge amount of background signal in the GFP
} } } channel that I normally don't see- so much so that I could not see
} } } the actual fluorescence from the sample. The fluorescence illuminator
} } } is a 470nm peak LED (Thorlabs) using a Chroma 49002 GFP filter set.
} } } The transmitted light source is a white LED (Thorlabs). I tracked
} } } the problem down to reflected light coming through the system from
} } } the transmitted light pathway. It turns out that normally, I have a
} } } phase ring in place in the condenser and so the excitation light
} } } coming through the sample is absorbed and does not cause a problem.
} } } However, I had moved the phase ring out of the path that day, and
} } } that led to light going through the condenser, up to the collimation
} } } optic in front of the white light LED tha! t I use for transmitted
} } } light imaging, reflecting off that optic and then going back down
} } } through the dichroic and emission filter flooding the image with
} } } green light. I am surprised that so much light could be reflecting
} } } off the collimation lens, but I am even more surprised that that
} } } light could get through both the dichroic and the emission filter.
} } } Any thoughts on why this is happening and the best way to solve it?
} } } It is not a huge problem because I can simple keep the phase ring in
} } } place, but I want to understand what is happening. Would
} } } anti-reflection coatings on the collimation optic fix the problem?
} } } Dave
} } }
} } } Dr. David Knecht Department of Molecular and Cell Biology Co-head
} } } Flow Cytometry and Confocal Microscopy Facility U-3125 91 N.
} } } Eagleville Rd. University of Connecticut Storrs, CT 06269
} } } 860-486-2200 860-486-4331 (fax)
} } }
} } }
} } }
} } }
} }
} } --
} } Aryeh Weiss
} } School of Engineering
} } Bar Ilan University
} } Ramat Gan 52900 Israel
} }
} } Ph: 972-3-5317638
} } FAX: 972-3-7384051
} }
}
} Dr. David Knecht
} Department of Molecular and Cell Biology
} Co-head Flow Cytometry and Confocal Microscopy Facility
} U-3125
} 91 N. Eagleville Rd.
} University of Connecticut
} Storrs, CT 06269
} 860-486-2200
} 860-486-4331 (fax)
}
}
}
}
} ==============================Original Headers==============================
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==============================Original Headers==============================
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10, 40 -- Subject: Re: [Microscopy] Re: Fluorescence reflection problem
10, 40 -- From: Rosemary White {rosemary.white-at-csiro.au}
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10, 40 -- Thread-Topic: [Microscopy] Re: Fluorescence reflection problem
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From: fedamann-at-gmail.com
Date: Wed, 8 Dec 2010 18:32:15 -0600
Subject: [Microscopy] viaWWW: Anthropology lab seeking a dissecting microscope

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Email: fedamann-at-gmail.com
Name: Franklin Damann

Organization: CMP

Title-Subject: [Filtered] Anthropology lab seeking a dissecting microscope

Message: The forensic anthropology laboratory of the Commission on
Missing Persons in Cyprus is seeking a dissecting microscope to
assist with their analytical casework. Any information on where the
lab may obtain a used model at an extremely reasonable price would be
greatly appreciated.

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From: rhunt-at-ucla.edu
Date: Wed, 8 Dec 2010 18:32:45 -0600
Subject: [Microscopy] viaWWW: Where can I get 2" steel samples polished in LA area?

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Email: rhunt-at-ucla.edu
Name: Ryan Hunt

Organization: University of California, Los Angeles

Title-Subject: [Filtered] Where can I get 2" steel samples polished in LA area?

Message: I have six two-inch discs (roughly 3/4" thick) that I need
to polish to a 0.5micron finish. I would do it by hand myself, but I
have so far been unable to maintain an appropriate level of flatness
to the surfaces. Does anyone know where I could get this type of
work done in the Los Angeles area?

Thanks for any help you can provide.

-Ryan

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8, 22 -- Subject: viaWWW: Where can I get 2" steel samples polished in LA area?
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From: ce-at-personifysearch.com
Date: Wed, 8 Dec 2010 18:33:39 -0600
Subject: [Microscopy] viaWWW: Position Opening: Product Manager

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Email: ce-at-personifysearch.com
Name: Christy Edwards

Organization: Personify

Title-Subject: [Filtered] Position Opening: Product Manager

Message: The Company:
Our client is a leading developer and producer of innovative high-tech
precision optics systems for the analysis of microstructures. As one of
the market leaders in each of the fields of Microscopy, Confocal Laser
Scanning Microscopy, Imaging Systems, Specimen Preparation and Medical
Equipment. Comprising nine manufacturing facilities in seven countries,
sales and service companies in 20 countries and an international network
of dealers, the company is represented in over 100 countries.

The Opportunity:
The company currently has an opening for a Product Manager to be based in
Chicago IL . All applicants must not be adverse to travel, as this is a
position that may require you to travel when necessary.

Base: $65-85k
Other: Full benefits - 401k program/matching

Primary Responsibilities:
This role will be responsible for the success (share, growth,
profitability and launch) of core product lines within multiple market
segments. The Product Manager will own the development and management of
the product life cycle and the training of commercial resources on product
features, benefits and value.

Additional Responsibilities:
- Enabling Wins: Negotiations, Demos, Quotes, Special Customer Requests,
Trade Shows and Seminars, Win/Loss Management
- Launching Products: Sales Tools, Product Training, Beta Testing, Pilots,
Product VOCs, Launch Planning
- Post Sales Support: Technical Support, Sales Processes, Product
Performance, Technology Assessment
- Managing Product: Innovation, Ramp Down, Forecasting, Lifecycle
Management, Collateral, Sales Tools, Transactional/Product Pricing

Education and Experience Required:
BA/BS in Life Sciences or equivalent is required. 2 years product
management experience in a related discipline/field is required. An
understanding of Medical/Clinical, Life Science Research or Industrial
marketplace is desired. The Product Manager must demonstrate success in
developing products within the optical instrumentation or microscopy
industry. A strong understanding of selling, quoting and working with
sales to enable wins is required. Strong leadership ability, strong
communication skills and expert computer skills are required.

If you meet and/or exceed the experience criteria, please submit your
resume by visiting www.personifysearch.com, clicking "Current Searches"
and then clicking on "Diagnostics" to view all listed openings with our
client. We wish everyone the best of luck. Unfortunately only qualified
candidates will be considered.


Christy Edwards
Sr. e-Recruitment Consultant
Personify
5020 Weston Parkway Suite 315
Cary NC 27513
www.personifysearch.com



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16, 22 -- Subject: viaWWW: Position Opening: Product Manager
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From: kraftpiano-at-gmail.com
Date: Wed, 8 Dec 2010 18:59:09 -0600
Subject: [Microscopy] Part needed for Hummer I sputter system

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hello everyone, I hope the holiday and academic final exam season is treating you well.

I have an old Hummer I sputter system, and the piece that holds the target has gotten lost after a move. I was wondering if someone either has this metal piece that is extra, or has one that can be measured to make a new one. It's not the whole top piece, just the metal stub that screws onto the high voltage terminal inside the lid.

Any info appreciated.

Thanks,

Justin A. Kraft

==============================Original Headers==============================
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5, 38 -- From: Justin Kraft {kraftpiano-at-gmail.com}
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5, 38 -- Subject: Part needed for Hummer I sputter system
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From: john.mitchels-at-gmail.com
Date: Thu, 9 Dec 2010 03:48:27 -0600
Subject: [Microscopy] Jan 2011, Imaging Conference, Bath. UK

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Dear List

We would like to invite you to a FREE one day conference at the
University of Bath on Wednesday 12th January 2011; a programme is
below.

The aims of the conference are to highlight some current techniques
and applications in microscopy and analysis. To bring together
researchers from different disciplines who use microscopy in their
research from the south west and south Wales areas. This is in the UK,
but anyone want to attend is welcome from outside the south west and
Wales.
We are encouraging groups and postgraduates to use this event as an
opportunity to meet others interested in microscopy and to display a
poster about their research. Poster prizes have been provided by our
industrial sponsors. There will be a trade exhibition involving
various
microscopy companies. Please see this link for more details:
http://www.bath.ac.uk/mas/conference/index.html

If you would like to attend please email us at mas-at-bath.ac.uk. Please
feel free to pass this information on to anyone you think may be
interested. For accommodation and transport please see the main
university travel page here http://www.bath.ac.uk/transport/

Regards
John Mitchels
University of Bath, UK
__________________________________________________________________

Microscopy & Analysis Conference 2011
Wednesday 12th January 2011
Venue: The University of Bath – 3 West North 2.
Programme:
09:00-10:00 Registration - coffee/tea available in ICIA art gallery
10:00-10:05 Welcome Sue Wonnacott – Chair of the MAS Management Committee
10:05-10:50 (Luminescence Imaging)“Imaging the cause and effect of
Ca2+ oscillations in eggs” Karl Swann - Cardiff University, School of
Medicine
10:50-11:20 (SPM/AFM) "New developments in AFM: from quantitative
materials characterisation to molecular recognition" Drew Murray –
Veeco Instruments
11:20-12:00 Break - coffee/tea in ICIA art gallery trade exhibits in 3WN 3.7
12:00-12:45 (FESEM/TEM & Raman Spectroscopy) “TEM and SEM of liquids
inside carbon nanotubes”  Davide Mattia – University of Bath,
Department Chemical Engineering
12:45-14:00 LUNCH BREAK – posters & trade exhibits in 3WN 3.7
14:00-14:45 (FIB/SEM,SBF/SEM, CLSM) “Imaging transient events at
subcellular resolution in whole organisms using correlative light and
volume EM” Lucy Collinson - Cancer Research UK, Lincoln's Inn Fields,
London.
14:45-15:30 (HRTEM/EELS) “Correlative Microscopy of
Nanomaterial-Tissue Interfaces”  Alexandra Porter - Imperial College,
London Centre for Nanotechnology.
15:30-15:50 Break - coffee/tea in ICIA art gallery trade exhibits in 3WN 3.7
15:50-16:35 (TEM/FESEM) "Nanomaterials: Hard and Soft Matters" Adam
Perriman – University of Bristol, Department Chemistry
16:35-17:15 (CLSM) “Macrophage migration and chemotaxis in Drosophila
embryos”  Will Wood - University of Bath, Department Biology &
Biochemistry
X-from 17:15 Wine networking reception - poster prize awards & trade
exhibits Registration is free – to register email:
U.J.Potter-at-bath.ac.uk Posters welcome


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7, 33 -- Subject: Jan 2011, Imaging Conference, Bath. UK
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From: oshel1pe-at-cmich.edu
Date: Thu, 9 Dec 2010 07:10:19 -0600
Subject: [Microscopy] Fwd: Ask-A-Microscopist Calibrating an EVEX EDS

Contents Retrieved from Microscopy Listserver Archives
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} Date: Wed, 8 Dec 2010 15:12:50 -0800 (PST)
} From: Pat McCurdy {patrick.mccurdy-at-colostate.edu}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Wednesday, December
} 08, 2010 at 03:12:43 PM.
}
} realname - Pat McCurdy
} Email - patrick.mccurdy-at-colostate.edu
} ORGANIZATION - Colorado State University
} EDUCATION - Graduate College
} LOCATION - Fort Collins
} SUBJECT_OF_QUESTION - EVEX EDS alignment issues
} QUESTION - I need some help with question about calibrating the EVEX
} EDS system. I have called EVEX a couple of times and they have been
} helpful, but they remind me that we are not under service contract.
} This is pretty simple problem possibly, but I need a little help.
}
} Thanks,
} Pat McCurdy
} Research Scientist
} Colorado State University
}
--
***************************************************************************************
Forwarded from "Ask a Microscopist"
Please remember that the person asking the question is likely
not a member the listserver, and
**any reply should go directly to the poster**
as well as to the list.
Using the "reply" function in your email does *not* send your answer
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Please copy their email address from their question.
****************************************************************************************

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1, 23 -- From: Philip Oshel {oshel1pe-at-cmich.edu}
1, 23 -- Subject: Fwd: Ask-A-Microscopist Calibrating an EVEX EDS
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From: ehaller-at-health.usf.edu
Date: Thu, 9 Dec 2010 07:31:32 -0600
Subject: [Microscopy] viaWWW: Anthropology lab seeking a dissecting microscope

Contents Retrieved from Microscopy Listserver Archives
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Franklin,

Two excellent places to look for used microscopes would be the following sources:


http://www.labx.com/
http://www.dotmed.com/

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: LSA 119
________________________________________
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Email: fedamann-at-gmail.com
Name: Franklin Damann

Organization: CMP

Title-Subject: [Filtered] Anthropology lab seeking a dissecting microscope

Message: The forensic anthropology laboratory of the Commission on
Missing Persons in Cyprus is seeking a dissecting microscope to
assist with their analytical casework. Any information on where the
lab may obtain a used model at an extremely reasonable price would be
greatly appreciated.

Login Host: 212.31.115.66
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From: ehaller-at-health.usf.edu
Date: Thu, 9 Dec 2010 07:57:36 -0600
Subject: [Microscopy] Resins & difficult-to-infiltrate algae & animal spores

Contents Retrieved from Microscopy Listserver Archives
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Hi, Giselle,

I find myself in a similar situation to yours. I'm a mammalian biologist that suddenly needs to process and embed algae for someone, and am having trouble with fixation and embedding. I also am not finding a good protocol. If someone sends you a good protocol, would you mind sharing it with me? I would appreciate it! I have a sample that I'm currently giving a try at. I started fixation with 4% paraformaldehyde/2.5% glutaraldehyde at 37 degrees C, and cooled the sample from there in the refrigerator. I left the sample fix over the weekend, then buffer rinsed overnight, and osmicated for 2 hours at room temp in 1% osmium. I rinsed for another hour in buffer, then started an acetone dehydration, 15 minutes each in 35%, 50%, 70%, and 95% acetone, 4 changes of 100% at 15 minutes each, then 2 changes of 100% propylene oxide. I spent hours gradually going through P.O.:resin infiltration, thinking that I would get good infiltration, but the algae floated in a 3:1 mix of resin:P.O., so I switched to acetone and resin, thinking that maybe I hadn't fully dehydrated the algae. I left the algae overnight in 50:50 acetone:resin at room temp, then went to 3:1 resin:acetone the next day and the algae still floated. I just continued on to higher resin concentration. I went to 100% resin for a day, then overnight in the refrigerator in resin. Now, I'm going a second day in resin on a rotator, then into the 70 degree oven at the end of the day. I don't know what else to try. I don't have access to an E.M. grade microwave, although I've heard that it is the best way to hndle these tough samples. If you hear of a better way, would you send it along? Thank you, and Merry Christmas!

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: LSA 119
________________________________________
X-from: gw265-at-cam.ac.uk [gw265-at-cam.ac.uk]
Sent: Tuesday, December 07, 2010 11:10 PM
To: Haller, Edward

Hi all

does anyone have specific advice for what to do with
difficult-to-infiltrate single cells? I'm looking for recommendations on
resins and on time schedules for infiltration and polymerisation.

I'm working with unicellular eukaryotic algae (2-5 microns, cell wall about
100nm thick) and myxozoan spores (10 microns, spore wall about 500nm
thick). Don't have positive evidence for what the walls are made of, but
it's not obviously mineralised. Both are very susceptible to poor
infiltration and extracted-looking cytoplasm. The literature is
uninformative about how long the good published examples have been
infiltrated.

Does "the longer, the better" apply here? Or will that make the extraction
worse? Or wouldn't it matter on the timescale of a week in resin?
Unfortunately with the EM unit closing for Christmas, and the fact the
myxozoan spores come from an endangered species, I don't have the option of
trying different lengths of time to see what's best. I guess if people were
really adamant that longer is definitely better, I could leave them
infiltrating until the unit opens again on the 5th January...

thanks

Giselle

Dr Giselle Walker
University of Cambridge UK
&
University of Sydney, Australia

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From: david.knecht-at-uconn.edu
Date: Thu, 9 Dec 2010 15:51:18 -0600
Subject: [Microscopy] Re: Fluorescence reflection problem solved

Contents Retrieved from Microscopy Listserver Archives
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Jim Beacher from CoolLED actually nailed this one. It was not the collimation optic, but the LED chip itself. It looks like what is happening is that the excitation light (470nm) is passed through the specimen, up to the collimation optic of hte white LED, which is focusing it onto the LED. The LED chip is coated with a phosphor that then emits a wide spectrum of light. That light emitted by the chip is then being sent back down through the transmitted light pathway and the green portion is being allowed through the GFP emission filter, swamping out the actual fluorescence signal. The simplest solution is to keep the phase ring in place in the condenser which blocks this light in both directions. Not a problem I would ever have dreamed of in advance. Thanks for the help and suggestions. Dave

On Dec 8, 2010, at 5:17 PM, Rosemary.White-at-csiro.au wrote:

}
}
}
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} Haven't been following this thread, but in the new Zeiss upright compound
} fluorescence microscopes, there is so much reflection from the top element
} of the condenser (why this problem has arisen now, in the very latest
} generation of microscopes is an open question), that you have to remove it
} from the light path when doing fluorescence. There are similar reflection
} problems in the fluorescence dissectors, but to a much lesser extent.
} Perhaps try this - rotating the top element out of the way? It does mean
} you then have to merge two images if you want to overlay fluorescence with
} DIC, which is often impossible with moving fluorescence objects.
}
} cheers,
} Rosemary
}
} Dr Rosemary White
} CSIRO Plant Industry
} GPO Box 1600
} Canberra, ACT 2601
} Australia
}
} T 61 2 6246 5475
} F 61 2 6246 5334
} E rosemary.white-at-csiro.au
}
}
}
} On 9/12/10 3:44 AM, "david.knecht-at-uconn.edu" {david.knecht-at-uconn.edu} wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------------
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} }
} } With the phase ring in place to eliminate the reflection, I tried with and
} } without the exciter and you need the exciter to get normal fluorescence
} } images. Without the exciter the background is too high. The LED profile
} } (M470 at link below) makes it look like there should be no signal to leak
} } through the dichroic and emission filters, but there is. The reflection
} } problem with the exciter in place and no phase ring actually causes worse
} } background than the LED with no exciter and no reflection. Dave
} }
} } http://www.thorlabs.com/images/TabImages/MxxxL2_spectra_dwg-LRG_dwg.png
} }
} } On Dec 8, 2010, at 10:43 AM, Aryeh Weiss wrote:
} }
} } } Dear David,
} } }
} } } Are you using the excitation filter of that Chroma set, or are you
} } } depending on the LED being narrow bandwidth? If you are not filtering
} } } the LED, the nit can have enough light over 500nm to produce this result.
} } }
} } } Best regards,
} } } --aryeh
} } }
} } } On 12/8/10 5:02 PM, david.knecht-at-uconn.edu wrote:
} } } } ----------------------------------------------------------------------------
} } } }
} } } }
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} } } }
} } } } I have an LED Illumination system that I have built for my Nikon TI
} } } } microscopes and have an issue that I don't understand that perhaps
} } } } someone else has insight on. I was looking at some samples recently
} } } } and found that I had a huge amount of background signal in the GFP
} } } } channel that I normally don't see- so much so that I could not see
} } } } the actual fluorescence from the sample. The fluorescence illuminator
} } } } is a 470nm peak LED (Thorlabs) using a Chroma 49002 GFP filter set.
} } } } The transmitted light source is a white LED (Thorlabs). I tracked
} } } } the problem down to reflected light coming through the system from
} } } } the transmitted light pathway. It turns out that normally, I have a
} } } } phase ring in place in the condenser and so the excitation light
} } } } coming through the sample is absorbed and does not cause a problem.
} } } } However, I had moved the phase ring out of the path that day, and
} } } } that led to light going through the condenser, up to the collimation
} } } } optic in front of the white light LED tha! t I use for transmitted
} } } } light imaging, reflecting off that optic and then going back down
} } } } through the dichroic and emission filter flooding the image with
} } } } green light. I am surprised that so much light could be reflecting
} } } } off the collimation lens, but I am even more surprised that that
} } } } light could get through both the dichroic and the emission filter.
} } } } Any thoughts on why this is happening and the best way to solve it?
} } } } It is not a huge problem because I can simple keep the phase ring in
} } } } place, but I want to understand what is happening. Would
} } } } anti-reflection coatings on the collimation optic fix the problem?
} } } } Dave
} } } }
} } } } Dr. David Knecht Department of Molecular and Cell Biology Co-head
} } } } Flow Cytometry and Confocal Microscopy Facility U-3125 91 N.
} } } } Eagleville Rd. University of Connecticut Storrs, CT 06269
} } } } 860-486-2200 860-486-4331 (fax)
} } } }
} } } }
} } } }
} } } }
} } }
} } } --
} } } Aryeh Weiss
} } } School of Engineering
} } } Bar Ilan University
} } } Ramat Gan 52900 Israel
} } }
} } } Ph: 972-3-5317638
} } } FAX: 972-3-7384051
} } }
} }
} } Dr. David Knecht
} } Department of Molecular and Cell Biology
} } Co-head Flow Cytometry and Confocal Microscopy Facility
} } U-3125
} } 91 N. Eagleville Rd.
} } University of Connecticut
} } Storrs, CT 06269
} } 860-486-2200
} } 860-486-4331 (fax)
} }
} }
} }
} }
} } ==============================Original Headers==============================
} } 8, 21 -- From david.knecht-at-uconn.edu Wed Dec 8 10:38:45 2010
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} 10, 40 -- From prvs=9517bc019=Rosemary.White-at-csiro.au Wed Dec 8 16:17:30 2010
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Dr. David Knecht
Department of Molecular and Cell Biology
Co-head Flow Cytometry and Confocal Microscopy Facility
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)




==============================Original Headers==============================
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From: bschneid-at-fhcrc.org
Date: Thu, 9 Dec 2010 15:53:04 -0600
Subject: [Microscopy] viaWWW: TEM Tomography

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Email: bschneid-at-fhcrc.org
Name: Bobbie Schneider

Organization: Fred Hutchinson Cancer Research Center

Title-Subject: [Filtered] RE: TEM Tomography

Message: I would like to know what type of room temperature TEM
tomography software people are using and any comments about the
software would be appreciated.

Thank you,
Bobbie Schneider
FHCRC, EM Resource

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From: rosemary.white-at-csiro.au
Date: Thu, 9 Dec 2010 17:05:18 -0600
Subject: [Microscopy] Resins & difficult-to-infiltrate algae & animal

Contents Retrieved from Microscopy Listserver Archives
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Hi Ed,

As I mentioned to Giselle (forgot to cc to list), I am familiar with Monica
Schoenwaelder's work on brown algal zygotes, and her search for a protocol
that would preserve phenolic vesicles - physodes, intact for TEM.

Here's an extract from one of Monica's papers ­ note the slow infiltration:
http://onlinelibrary.wiley.com/doi/10.1055/s-2000-9178/abstract

For electron microscopy, eggs, zygotes and embryos were fixed in a
glutaraldehyde/paraformaldehyde fixative (Schoenwaelder and Clayton,
1998a[37], b[38]) for 2­3 h. They were then rinsed three times in sodium
cacodylate buffer (2% sodium chloride and 0.1% calcium chloride in 0.1M
sodium cacodylate buffer), and post-fixed in 1% osmium tetroxide in buffer
for 2 h. Specimens were rinsed three times in distilled water to eliminate
excess osmium tetroxide and then dehydrated through a graded acetone series
of 10% increments, before three changes in 100% acetone. The specimens were
infiltrated very gradually (over 2 weeks) with Spurr¹s resin (Spurr,
1969[42]) (medium/hard mixture) before polymerization at 60 C in plastic
beem capsules.

Monica also mentioned that she started her infiltrations with 1% resin
increments, perhaps increasing from 1% to 5% over day one, then 5% to 10%
day two, then increased in larger steps in the middle range of
concentrations, and then slowed down again after reaching 90% resin. (This
is what I remember, you could check her earlier papers for details - our
library doesn't get Phycologia any more.) The brown algal zygotes are not
only quite dense but also have surprisingly impermeable walls - the wall
pores allow rather slow resin monomer permeation, but the small solvent
molecules can escape quickly, of course, and if the gradient across the wall
is too great, the zygotes crumple inwards.

A further trick, outlined by Geoff Wasteneys & co., is to freeze the cells
briefly, then thaw - to make a few cracks in the cell wall. See Wasteneys
et al. 1997. Freeze-shattering: a simple and effective method for
pemeabilizating higher plant cell walls. J. Microsc. 188, pp. 51­61. Can't
copy the bit of the method because these pdfs don't allow it. Essentially,
you freeze the tissue between two slides in liquid nitrogen then press down
gently while frozen, then thaw. This works pretty well, but requires a bit
of practise.

cheers,
Roseamry

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au





On 10/12/10 1:01 AM, "ehaller-at-health.usf.edu" {ehaller-at-health.usf.edu}
wrote:

}
}
}
} ----------------------------------------------------------------------------
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}
} Hi, Giselle,
}
} I find myself in a similar situation to yours. I'm a mammalian biologist
} that suddenly needs to process and embed algae for someone, and am having
} trouble with fixation and embedding. I also am not finding a good protocol. If
} someone sends you a good protocol, would you mind sharing it with me? I would
} appreciate it! I have a sample that I'm currently giving a try at. I started
} fixation with 4% paraformaldehyde/2.5% glutaraldehyde at 37 degrees C, and
} cooled the sample from there in the refrigerator. I left the sample fix over
} the weekend, then buffer rinsed overnight, and osmicated for 2 hours at room
} temp in 1% osmium. I rinsed for another hour in buffer, then started an
} acetone dehydration, 15 minutes each in 35%, 50%, 70%, and 95% acetone, 4
} changes of 100% at 15 minutes each, then 2 changes of 100% propylene oxide. I
} spent hours gradually going through P.O.:resin infiltration, thinking that I
} would get good infiltration, but the algae floated in a 3:1 mix of resin:!
} P.O., so I switched to acetone and resin, thinking that maybe I hadn't fully
} dehydrated the algae. I left the algae overnight in 50:50 acetone:resin at
} room temp, then went to 3:1 resin:acetone the next day and the algae still
} floated. I just continued on to higher resin concentration. I went to 100%
} resin for a day, then overnight in the refrigerator in resin. Now, I'm going a
} second day in resin on a rotator, then into the 70 degree oven at the end of
} the day. I don't know what else to try. I don't have access to an E.M. grade
} microwave, although I've heard that it is the best way to hndle these tough
} samples. If you hear of a better way, would you send it along? Thank you, and
} Merry Christmas!
}
} Ed
}
} Edward Haller, Lab Manager
} University of South Florida
} Integrative Biology Department
} Electron Microscopy Core
} SCA 110
} 4202 East Fowler Avenue
} Tampa, FL 33620
} 813-974-2676
} ehaller-at-usf.edu
} Office: LSA 119
} ________________________________________
} X-from: gw265-at-cam.ac.uk [gw265-at-cam.ac.uk]
} Sent: Tuesday, December 07, 2010 11:10 PM
} To: Haller, Edward
} Subject: [Microscopy] Resins & difficult-to-infiltrate algae & animal spores
}
} ----------------------------------------------------------------------------
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} Hi all
}
} does anyone have specific advice for what to do with
} difficult-to-infiltrate single cells? I'm looking for recommendations on
} resins and on time schedules for infiltration and polymerisation.
}
} I'm working with unicellular eukaryotic algae (2-5 microns, cell wall about
} 100nm thick) and myxozoan spores (10 microns, spore wall about 500nm
} thick). Don't have positive evidence for what the walls are made of, but
} it's not obviously mineralised. Both are very susceptible to poor
} infiltration and extracted-looking cytoplasm. The literature is
} uninformative about how long the good published examples have been
} infiltrated.
}
} Does "the longer, the better" apply here? Or will that make the extraction
} worse? Or wouldn't it matter on the timescale of a week in resin?
} Unfortunately with the EM unit closing for Christmas, and the fact the
} myxozoan spores come from an endangered species, I don't have the option of
} trying different lengths of time to see what's best. I guess if people were
} really adamant that longer is definitely better, I could leave them
} infiltrating until the unit opens again on the 5th January...
}
} thanks
}
} Giselle
}
} Dr Giselle Walker
} University of Cambridge UK
} &
} University of Sydney, Australia
}
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From: oshel1pe-at-cmich.edu
Date: Fri, 10 Dec 2010 12:47:09 -0600
Subject: [Microscopy] Fwd: Ask-A-Microscopist Artemia TEM

Contents Retrieved from Microscopy Listserver Archives
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} Date: Fri, 10 Dec 2010 10:33:39 -0800 (PST)
} From: Gigi Kemalyan {gkemalyan-at-yahoo.com}
} To: oshel1pe-at-cmich.edu
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Friday, December 10,
} 2010 at 10:33:30 AM.
}
} realname - Gigi Kemalyan
} Email - gkemalyan-at-yahoo.com
} ORGANIZATION - San Joaquin Delta College
} EDUCATION - Undergraduate College
} LOCATION - Stockton, CA, USA
} SUBJECT_OF_QUESTION - Identifying Artemia Ultrastructure
} QUESTION - Dear Microscopists,
}
} I am an undergraduate student in San Joaquin Delta College's
} Electron Microscopy program. My partners and I are working on a
} special project for a biological specimen preparation course and
} need some expert help!
}
} We prepared Artemia salina eggs, nauplii, and adults for TEM viewing
} using two different fixatives, KMnO4 and Karnovsky's. The samples
} were embedded in Spurr's and post stained with uranyl acetate and
} lead citrate, and we got some pretty good pictures. However, we are
} having trouble identifying the ultrastructures and are hoping the
} friendly folks on the list server can help us.
}
} A search for books on the biology/physiology of Artemia came up with
} several good possibilities to use as references, but all of these
} volumes are checked out at the university libraries in our area
} (including UC Davis), so we were not able to consult them. Online
} searches have proved equally fruitless. We also have not been able
} to find exact matches to what we are seeing in our pictures in other
} books with TEM images that are not specifically about brine shrimp.
} We would sincerely appreciate your help in figuring out *what* it is
} we have photographed!
}
} Below is a link to a Flickr account I have set up for our images.
} Please take a look and leave comments on the images! [And if we get
} an A on the project, we will give you credit as well!] ;)
}
} http://www.flickr.com/photos/56947013-at-N08/
}
} Thank you,
}
} Gigi Kemalyan
} Nan Ny
} Kelly Wagner
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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From: oshel1pe-at-cmich.edu
Date: Fri, 10 Dec 2010 13:21:22 -0600
Subject: [Microscopy] Fwd: Ask-A-Microscopist Scherzer focus vs optimum underfocus

Contents Retrieved from Microscopy Listserver Archives
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} Date: Fri, 10 Dec 2010 11:14:26 -0800 (PST)
} From: Bernard Schreurs {bschreurs-at-brni.org}
} Subject: Ask-A-Microscopist
}
} Below is the result of your form, submitted on Friday, December 10,
} 2010 at 11:14:17 AM.
}
} realname - Bernard Schreurs
} Email - bschreurs-at-brni.org
} ORGANIZATION - West Virginia University
} EDUCATION - Graduate College
} LOCATION - Morgantown, WV
} SUBJECT_OF_QUESTION - Schertzer Focus
} QUESTION - I have been reading that most microscopes can be preset
} on "Schertzer focus" and there is discussion of "optimum
} underfocus". There are also significant changes in contrast
} associated with changes in focus that can lead to misinterpreting
} images.
}
} I would like to hear your comments on the these topics.
}
--
***************************************************************************************
Forwarded from "Ask a Microscopist"
Please remember that the person asking the question is likely
not a member the listserver, and
**any reply should go directly to the poster**
as well as to the list.
Using the "reply" function in your email does *not* send your answer
to the person asking the question.
Please copy their email address from their question.
****************************************************************************************

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From: William.F.Tivol-at-aero.org
Date: 12/10/2010 11:28 AM
Subject: [Microscopy] Fwd: Ask-A-Microscopist Scherzer focus vs

Contents Retrieved from Microscopy Listserver Archives
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Dear Bernard,
First, I recommend that you read any good reference book on
transmission electron microscopy. The section on phase contrast
microscopy will describe Schertzer focus--if there is no such section in
the book you have chosen, look for another book. A simple definition of
Schertzer focus is that it is the point where the effects of spherical
aberration and defocus offset each other to the maximum extent (this
always happens at underfocus). This allows information to be transmitted
from the specimen to the image in phase at the highest spatial frequency
for which there are no phase reversals at lower spatial frequencies. High
spatial frequency corresponds to small details, and the lack of phase
reversal means that interpretation of the image is pretty straightforward
(there is still some loss of intensity at high spatial frequency, that is,
a loss of contrast for small details, but bright things stay bright and
dark things stay dark). Optimum defocus (also always at underfocus) is
where the effect of spatial incoherence (the effect of a source of
electrons that is of finite size) of the beam is minimal. This means that
the envelope function, which damps out contrast, is as constant as
possible to high spatial frequencies. In practical terms, this means that
the signal-to-noise ratio is highest for details of all sizes down to the
smallest resolvable. In other words, it is the defocus for which details
of all sizes are equally visible (as nearly as possible). For phase
contrast imaging, the information in the specimen that is transmitted by
the microscope to the image can either be in phase or out of phase. In
the latter case, the appearance of the image will be different from the
specimen, giving the possibility of misinterpretation. One can compensate
for the out-of-phase character of the information at spatial frequencies
where the phase is reversed by dividing out the effect of the contrast
transfer function--look it up in the book--to get a more simply
interpretable image.
Yours,
Bill



X-from: oshel1pe-at-cmich.edu
To: William.F.Tivol-at-aero.org

}
} realname - Bernard Schreurs
} Email - bschreurs-at-brni.org
} ORGANIZATION - West Virginia University
} EDUCATION - Graduate College
} LOCATION - Morgantown, WV
} SUBJECT_OF_QUESTION - Schertzer Focus
} QUESTION - I have been reading that most microscopes can be preset
} on "Schertzer focus" and there is discussion of "optimum
} underfocus". There are also significant changes in contrast
} associated with changes in focus that can lead to misinterpreting
} images.
}
} I would like to hear your comments on the these topics.
}


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From: ron.doole-at-materials.ox.ac.uk
Date: Sat, 11 Dec 2010 01:12:51 -0600
Subject: [Microscopy] TEM holder

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Hi Roger,

We have some JEOL double tilt holders which have jewelled pivot screws and bearings on the tilt gimbal. We have had experiences of these being damaged and we do repair them ourselves, obtaining the pivot screws from JEOL.

There are several causes of user damage that are avoidable.
It is important that the holder tip is lowered gently onto the support after the heavier end has been put down fully and that the tip is lifted first. This will avoid the gimbal taking the weight of the holder on the delicate pivot jewells.
It is important to ensure that the holder tip is sitting straight on the stand, that the support is directly below the gimbal and that the holder is not rotated on the stand. (Later support stands do square up the holder to ensure it is not rotated.)
The JEOL holder stand has a variable height centre support that is set at the factory but I have known these to become loose and users to adjust them incorrectly so that the weight of the holder is being held by the gimbal as opposed to the weight of the holder being taken by the stand with the gimbal support just underneath the gimbal and not taking any weight.
Users can try to overtighten the specimen screw ring which will rotate the gimbal and can break the pivot screws. This is a matter of 'feel' and there are a few people who are unable to be delicate, they ought not handle delicate items and it may be better to make alternative arrangements for their specimen loading.

It is best to take the above precautions and not to damage the holder in the first place but we have had enough problems with our holders that we now keep pivot screws in stock and replace them in-house. The screws are not cheap but we can get a holder back in action in a day or so provided that the bearing cup on the gimbal has not been damaged, we have not been able to source the bearing cups and have to replace the gimbal complete.

Apart from the delicate nature of the repair work it is relatively simple, basic instructions from memory are:
Carefully unclip the gimbal from the tilt mechanism so that the ginbal is free to rotate.
The original screws will have been sealed with epoxy resin to ensure that do not move from their set position. This must be dissolved with acetone before you can remove the old screw. We use a cotton bud to keep acetone only on the screw to be removed.
When the screw has been removed the tapped hole needs to be cleaned of any epoxy remains.
Fit the new screw such that the gimbal will just not drop under its own weight then back off the screw 5-10 deg rotation so that the gimbal is free to drop under its own weight but there is no side to side movement of the gimbal.
Put a little, and I mean a little, epoxy onto the top of the screw to keep it in the set position. Do not get it into the screw thread or it will be very difficult to remove for any future repairs.
Replace the gimbal into the tilt mechanism.
Check that the holder stand is still set to the correct height for the new screw(s).
Leave the epoxy overnight to harden before exposing it to the vacuum system.
If the above does not seem obvious to you then is it probably best that you do not attempt the repair.

We have been unable to source the pivot screws except from JEOL but we have developed a method of removing the old jewell from the screw and have made a new screw from one with a damaged jewell and one with a damaged screw head. We have managed to source the jewells seperately but we would have to buy them in bulk ( I think it was 20 minimum) so we would have to make several pivot screws for it to be economically viable. We hope not to have to repair that many holders.

regards,
Ron

Mr. Ron Doole Department of Materials
Senior Instrumentation Engineer. University of Oxford.
Phone +44 (0) 1865 273701 Parks Road. Oxford. OX1 3PH
Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
________________________________________
X-from: raristau-at-ims.uconn.edu [raristau-at-ims.uconn.edu]
Sent: 08 December 2010 21:55
To: Ron Doole

Hi All,

Does anyone know of a source for the small jeweled pivot-pins used to hold
specimen cup on double-tilt TEM holders? It seems that heavy-handed users in
our multi-user facility break these jewels every year or so, and I am not
satisfied with the hassle of sending out the holder for repair.

I am sure we are not the only ones to experience this. Does anyone have
hints for on-site repair?

Cheers
Roger

Roger A. Ristau, PhD
Electron Microscopy Specialist
Institute of Materials Science
97 North Eagleville Road
University of Connecticut
Storrs, CT 06269
vox: 860-486-5453
fax: 860-486-4745




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18, 30 -- Date: Sat, 11 Dec 2010 07:12:48 +0000
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From: germpore-at-sonic.net
Date: Sun, 12 Dec 2010 01:24:06 -0600
Subject: [Microscopy] Brands of plain glass slides

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I was wondering if anybody can explain what qualitative difference
exist, if any, between different brands of plain glass slides. For
example, over on the Ted Pella page ( http://www.tedpella.com/histo_html/slides.htm
), for a 2-box pack of 144 slides, the price varies from about $10
for the Pella Economy Slides, all the way up to $40 or $60+ for Gold
Seal or Corning slides.

My question is, is there any advantage or optical difference with the
more expensive slides? Corning advertises theirs as "water white"
glass, which I imagine implies something about the transmittance or
refractive index, but I'm not sure.

==============================Original Headers==============================
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From: PhillipsT-at-missouri.edu
Date: Sun, 12 Dec 2010 12:43:38 -0600
Subject: [Microscopy] Brands of plain glass slides

Contents Retrieved from Microscopy Listserver Archives
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Water-white glass is low iron and has 98-99% transmission. If you ever have looked at a slab of window glass on edge, you will have notice the edge looks green since the glass absorbs some red and blue light. I guess I have seen that same phenomenon when I have looked at a new box of microscope slides where they are packed together and the edges look green. I intend to compare my different brands of slides when I go into work later today to confirm that recollection. I wonder how important the effect is in microscopy for brightfield work. These seem characteristics that would be important for the coverslip glass but less so for the slide to me since the amount of incoming light that passes through glass is usually not limiting. The color balance of the light going through the glass is partly dependent on the power to the tungsten bulb which is why microscopes used to always have a button for fixing the power of the bulb when taking color slide film in the old days and one needed to take into account whether you were using daylight or indoor style film. Most digital cameras use white balance to eliminate this problem so I don't know if the glass absorption of some wavelengths is a significant issue for regular bright field work. On the coverslip side, getting all the light from fluorescent specimens would be beneficial but in this case both the excitation and emission light are passing through the coverslip. Tom




Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: germpore-at-sonic.net [mailto:germpore-at-sonic.net]
Sent: Sunday, December 12, 2010 1:25 AM
To: Phillips, Thomas E.

I was wondering if anybody can explain what qualitative difference
exist, if any, between different brands of plain glass slides. For
example, over on the Ted Pella page ( http://www.tedpella.com/histo_html/slides.htm
), for a 2-box pack of 144 slides, the price varies from about $10
for the Pella Economy Slides, all the way up to $40 or $60+ for Gold
Seal or Corning slides.

My question is, is there any advantage or optical difference with the
more expensive slides? Corning advertises theirs as "water white"
glass, which I imagine implies something about the transmittance or
refractive index, but I'm not sure.

==============================Original Headers==============================
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From: gary-at-gaugler.com
Date: Sun, 12 Dec 2010 20:10:22 -0600
Subject: [Microscopy] Re: Brands of plain glass slides

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

FWIW, I only use Corning 2947 LM slides. The big problem
is finding good cover slips that are not made in China.
These lousy units come pre-cracked. Wonderful. I pay more
for quartz slips.

Whatever the color transmittance is, I can correct it with
digital capture software light balance.

gary g.

Here comes the OoO deluge.


At 11:26 PM 12/11/2010, you wrote:



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From: smithj-at-winthrop.edu
Date: Mon, 13 Dec 2010 14:10:49 -0600
Subject: [Microscopy] SEM: Quorum Technologies and Energy Beam Sciences

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Anyone out there have recent contact with either of these two
companies? Are they still solvent?
Neither of them seems to answer e-mail or telephone messages, and I've
been trying for over a year.
Julian

--
Julian P.S. Smith III
Director, Winthrop Microscopy Facility
Dept. of Biology
Winthrop University
520 Cherry Rd.
Rock Hill, SC 29733

803-323-2111 x6427 (vox)
803-323-3448 (fax)
803-524-2347 (cell)
Research Website www.birdnest.org/smithj
Personal Website www.rociada-east.net


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From: jd-at-laddresearch.com
Date: Mon, 13 Dec 2010 14:48:02 -0600
Subject: [Microscopy] Re: SEM: Quorum Technologies and Energy Beam

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I would suggest you call Mike Nesta at Energy Beam Sciences at 1-800-992-9037.

Even though we are competitive companies they are very knowledgeable
and helpful.

Thanks,
John Arnott

Ladd Research
83 Holly Court
Williston, VT 05495

On-line Catalog: www.laddresearch.com

Telephone: 1-802-658-4961 (anywhere)
Toll Free 1-800-451-3406 (US)
Fax: 1-802-660-8859

e-mail: sales-at-laddresearch.com

At 03:19 PM 12/13/2010, you wrote:



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From: smithj-at-winthrop.edu
Date: Mon, 13 Dec 2010 15:05:29 -0600
Subject: [Microscopy] Re: SEM: Quorum Technologies and Energy Beam Sciences

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Thanks for the help, folks. Problem solved.
Julian



--
Julian P.S. Smith III
Director, Winthrop Microscopy Facility
Dept. of Biology
Winthrop University
520 Cherry Rd.
Rock Hill, SC 29733

803-323-2111 x6427 (vox)
803-323-3448 (fax)
803-524-2347 (cell)
Research Website www.birdnest.org/smithj
Personal Website www.rociada-east.net


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From: klivi-at-jhu.edu
Date: Mon, 13 Dec 2010 16:02:23 -0600
Subject: [Microscopy] 4th Annual FIB SEM Workshop in Baltimore

Contents Retrieved from Microscopy Listserver Archives
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The date is fast approaching for the 4th Annual FIB/SEM Workshop (an affiliate of the Mid-Atlantic Microbeam Analysis Society). This year it will be held in Mason Hall at Homewood Campus, Johns Hopkins University, Baltimore Friday, February 25th, 2011. The agenda is already shaping up to include many interesting talks from both the Scientific and Vendor communities. As has been in the past, the meeting is free, yet includes breakfast, lunch, and a Happy Hour afterwards! We are calling for talks and posters to contribute to the days sessions. Please take the time to go to our website www.fibsem.org and register using the "contact us" box. Please indicate in the comment area if you want to give a talk or poster and its title, and to let us know if you want to come to the Happy Hour. We look forward to seeing all our FIB friends again and sharing new insights with each other! If you know someone who didn't get this email and should, please pass it along.

You don't have to come from the area to attend! We've had people from all over the US and Europe attend. Hope to see you in Baltimore.

Sincerely,
The Organizing Committee

Ken Livi klivi-at-jhu.edu
Keana Scott keana.scott-at-nist.gov
Nabil Bassim nabil.bassim-at-gmail.com

|_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
Kenneth JT Livi, PhD
Director, The High-Resolution Analytical Electron Microbeam Facility
of the Integrated Imaging Center
Departments of Earth and Planetary Sciences and Biology
Olin Hall
3400 N Charles Street
Johns Hopkins University
Baltimore, Maryland 21218 USA
| | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| | | :|










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From: bigelow-at-umich.edu
Date: Mon, 13 Dec 2010 20:57:53 -0600
Subject: [Microscopy] RE: Jeweled pivot screws

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In regard to Roger Ristau's and Ron Dooley's discussion of the
replacement of damaged jeweled pivot screws in double-tilt specimen
holders: I wonder if the use of jeweled screws in the first place
is not a bit of an overkill. These devices do not rotate very
rapidly, nor through great amounts, and so I would think that
carefully polished metal pivot screws would serve as well. These
could be made rather easily by most machinists, and they would not be
brittle and easily broken.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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From: wolpensinger-at-isfh.de
Date: Tue, 14 Dec 2010 01:02:31 -0600
Subject: [Microscopy] worn tracball on S-4800 - your experiences?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi all,

After about 3 years the trackball-unit on our S-4800 is so heavily worn that we now use it by removing the ball and turning the inner rods by hand.

When asking for a replacement, the company seemed to have a hard time, and told us that the trackball unit does not have a part number, nor can it be replaced solely.
Meanwhile we recieved a quote, but surprisingly the delivery is sheduled in 90 days!! This gives me the impression that we might be the only users on the planet being troubled with a worn trackball?!

Does anybody have similar experiences?
Any recommendations?
I've naiively wondered if it really needs a trackball, or if a simple joystick could do the same job?

Thanks in advance,

Bettina





Bettina Wolpensinger
Institute for Solar Energy Research Hamelin (ISFH)
An-Institut der Leibniz Universität Hannover
Am Ohrberg 1
D-31860 Emmerthal
Germany
Tel: +49 5151 999 313
Fax: +49 5151 999 400
Email:
wolpensinger-at-isfh.de
Handelsregister: Amtsgericht Hannover HRB 100547
Geschäftsführer: Prof. Dr. Rolf Brendel
Vorsitzender des Aufsichtsrats: Ministerialrat Dr. Hans Schroeder


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From: kraftpiano-at-gmail.com
Date: Tue, 14 Dec 2010 01:35:00 -0600
Subject: [Microscopy] Re: worn tracball on S-4800 - your experiences?

Contents Retrieved from Microscopy Listserver Archives
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I remember my computer consulting days when trackballs were considered viable alternatives to computer mice...

Every now and then we had issues with trackballs (and mouse balls) not functioning properly, and the fix was usually quite simple. First, clean the ball thoroughly. Also clean the little rollers it sits on. Attached to those rollers is a wheel with a bunch of slots in it. As the wheel turns, the slots make and break an optical beam. If the cleaning alone doesn't do it, take the trackball unit apart so you can see the opto sensors. They should look like small black towers on either side of the slotted wheel. Take some compressed air and blow them out really well. Those two steps fixed 99.99% of trackball an mouse ball issues.

--Justin A. Kraft

"America believes in education; the average professor earns more money in a year than a professional athlete earns in a whole week." Evan Esar

On Dec 14, 2010, at 2:08 AM, wolpensinger-at-isfh.de wrote:

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} Hi all,
}
} After about 3 years the trackball-unit on our S-4800 is so heavily worn that we now use it by removing the ball and turning the inner rods by hand.
}
} When asking for a replacement, the company seemed to have a hard time, and told us that the trackball unit does not have a part number, nor can it be replaced solely.
} Meanwhile we recieved a quote, but surprisingly the delivery is sheduled in 90 days!! This gives me the impression that we might be the only users on the planet being troubled with a worn trackball?!
}
} Does anybody have similar experiences?
} Any recommendations?
} I've naiively wondered if it really needs a trackball, or if a simple joystick could do the same job?
}
} Thanks in advance,
}
} Bettina
}
}
}
}
}
} Bettina Wolpensinger
} Institute for Solar Energy Research Hamelin (ISFH)
} An-Institut der Leibniz Universität Hannover
} Am Ohrberg 1
} D-31860 Emmerthal
} Germany
} Tel: +49 5151 999 313
} Fax: +49 5151 999 400
} Email:
} wolpensinger-at-isfh.de
} Handelsregister: Amtsgericht Hannover HRB 100547
} Geschäftsführer: Prof. Dr. Rolf Brendel
} Vorsitzender des Aufsichtsrats: Ministerialrat Dr. Hans Schroeder
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7, 42 -- Subject: Re: [Microscopy] worn tracball on S-4800 - your experiences?
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From: dservando86-at-yahoo.com
Date: Tue, 14 Dec 2010 01:52:29 -0600
Subject: [Microscopy] TEM- images displaying liver decay

Contents Retrieved from Microscopy Listserver Archives
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Hello, I am an EM student working on a project for my advanced ultramicrotomy
class.  I chose to compare a freshly fixed sample compared to a sample that
underwent the process of decay for three days before fixation.  For the 3 day
old liver tissue sample, I could only find collagen fibers throughout my
sample.  Although, there is another structure throughout my images and I am
needing help with identifying what it is.  I was thinking it was fibroblast
cells or even elastic tissue, however, I could be completely off and it may just
be an artifact or contaminant of some sort????  Furthermore, I have yet to be
successful with my research in order to determine why the collagen fibers are
the only remaining structures during the decay process.  The only research I
have found so far, would be the increase of collagen fibers when the liver is
induced with hepatic fibrosis.  Could someone PLEASE enlighten me with some
direction for my research?? I can email you my images if needed.
Thanks in advance for any advice!
Donya S.





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From: benada-at-biomed.cas.cz
Date: Tue, 14 Dec 2010 03:21:38 -0600
Subject: [Microscopy] Re: worn tracball on S-4800 - your experiences?

Contents Retrieved from Microscopy Listserver Archives
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Hi Betina,
Some years ago we had to solve similar problem with trackball of our Aquasem SEM.
We have carefully disassembled the whole trackball. Under binocular we remove all hairs, debris
and dirt from roller (inner rod) pins with EM tweezers and sprayed some canned air on them. Next
we cleaned the pins with 50% propanol in water and sprayed them with canned air again . Finally
we gently touched the pins with toothpick dipped in lubricating oil.
After reassembling, our trackball started to work again.
I hope it might work for you.

Best regards Oldrich
--
Oldrich Benada
Institute of Microbiology
Acad. Sci. CR
Videnska 1024
CZ-142 20 Prague 4
Czech Republic


On Tuesday 14 of December 2010 08:05:03 you wrote:
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} Hi all,
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} After about 3 years the trackball-unit on our S-4800 is so heavily worn
} that we now use it by removing the ball and turning the inner rods by
} hand.
}
} When asking for a replacement, the company seemed to have a hard time, and
} told us that the trackball unit does not have a part number, nor can it be
} replaced solely. Meanwhile we recieved a quote, but surprisingly the
} delivery is sheduled in 90 days!! This gives me the impression that we
} might be the only users on the planet being troubled with a worn
} trackball?!
}
} Does anybody have similar experiences?
} Any recommendations?
} I've naiively wondered if it really needs a trackball, or if a simple
} joystick could do the same job?
}
} Thanks in advance,
}
} Bettina
}
}
}
}
}
} Bettina Wolpensinger
} Institute for Solar Energy Research Hamelin (ISFH)
} An-Institut der Leibniz Universität Hannover
} Am Ohrberg 1
} D-31860 Emmerthal
} Germany
} Tel: +49 5151 999 313
} Fax: +49 5151 999 400
} Email:
} wolpensinger-at-isfh.de
} Handelsregister: Amtsgericht Hannover HRB 100547
} Geschäftsführer: Prof. Dr. Rolf Brendel
} Vorsitzender des Aufsichtsrats: Ministerialrat Dr. Hans Schroeder
}
}
} ==============================Original
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==============================Original Headers==============================
5, 24 -- From benada-at-biomed.cas.cz Tue Dec 14 03:21:37 2010
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From: peter.heimann-at-uni-bielefeld.de
Date: Tue, 14 Dec 2010 09:15:26 -0600
Subject: [Microscopy] LM: lacZ-staining (beta-galactosidase) on paraffin sections ?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Colleagues,

has anybody performed successfully beta-galactosidase-staining on
de-paraffinized sections of fixed, paraffin embedded (mouse) tissue ?

If YES, thank you for a short informal answer.

regards,

Peter Heimann

--
====================================================

Dr. Peter Heimann Raum: W7-107 / Tel.: +49-(0)521-106-5628
Universitaet Bielefeld Fax: +49-(0)521-106-5654
"Zellbiologie / Cell Biology" W7-107
33501 Bielefeld, Germany www.uni-bielefeld.de/biologie/cellbio



==============================Original Headers==============================
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From: peter.heimann-at-uni-bielefeld.de
Date: Tue, 14 Dec 2010 09:23:21 -0600
Subject: [Microscopy] clairification: LM: lacZ-staining (beta-galactosidase) on paraffin

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Colleagues,

to specify my question:

"has anybody performed successfully beta-galactosidase-staining on
de-paraffinized sections of fixed, paraffin embedded (mouse) tissue ? "

I think of beta-galactosidase staining by the X-gal (chromogenic BCIP-)
method, NOT by using anti-galactosidase antibodies.


If YES, thank you for a short informal answer.

regards,

Peter Heimann

--
====================================================

Dr. Peter Heimann Raum: W7-107 / Tel.: +49-(0)521-106-5628
Universitaet Bielefeld Fax: +49-(0)521-106-5654
"Zellbiologie / Cell Biology" W7-107
33501 Bielefeld, Germany www.uni-bielefeld.de/biologie/cellbio



==============================Original Headers==============================
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11, 31 -- sections ?
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From: swalck-at-southbaytech.com
Date: Tue, 14 Dec 2010 10:18:37 -0600
Subject: [Microscopy] Jeweled pivot screws

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

When I was in graduate school, my advisor, John Hren, told me that his group
had made some special TEM rod holders. They fabricated the jewel surface by
taking a glass rod and heating it and stretching it so that it was very
thin. Then after they broke the glass strand, they heated the end and it
made a glass sphere that they could control the size by how much they heated
the end of the strand. When they got the size that they wanted, they broke
off the sphere. The little tip of glass that was attached to the strand
went into the hole in the end of the rod and they glued (epoxy??) it in the
end of the rod. He said it worked well. I made a rod for FIM samples and
used the technique on a holder for a JEOL 200 CX.

-Scott

Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA 92673

eMail: SWalck-at-SouthBayTech.com
Phone: 949-492-2600
Toll Free: 1-800-728-2233 (USA)
Fax: 949-492-1499

www.SouthBayTech.com

s==============================


==============================Original Headers==============================
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From: mmashore-at-vapop.ucsd.edu
Date: Tue, 14 Dec 2010 11:00:09 -0600
Subject: [Microscopy] LSM 510 software issue

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hello,

I am having an issue with LSM 510 version 4.2 SP1. The software isn't
recording the position of the stage into the database for any saved images.
While scanning a slide the position will be displayed, but once an image is
recorded and saved, the database always tells me that the x and y
coordinates are 0. If anybody has any experience with this or has any
suggestions I would appreciate the help.

Thanks,

Michael




==============================Original Headers==============================
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From: William.F.Tivol-at-aero.org
Date: Tue, 14 Dec 2010 11:31:10 -0600
Subject: [Microscopy] Other solvent for colloidal gold

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Lists,
I have a specimen that is very small and not too firmly attached
to its substrate to which I would like to add colloidal gold. I am very
concerned that the surface tension in a few-ul drop of aqueous colloidal
gold will detach the specimen, so I would like to transfer the gold to a
solvent or suspension medium that has much lower surface tension, such as
ethanol or acetone. I am not concerned that either of these solvents
would harm the specimen, but they might not give a good dispersal of the
gold. Has anyone had experience with how gold disperses in various
solvents? TIA for any help.
Yours,
Bill

==============================Original Headers==============================
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From: ehaller-at-health.usf.edu
Date: Tue, 14 Dec 2010 12:48:12 -0600
Subject: [Microscopy] Other solvent for colloidal gold

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi, Bill,

You didn't mention if you are doing immunolabeling or not, which would change your approach to the application of the gold particles. If you are immunolabeling, you might try increasing the concentration of Tween 20 in your solution to lower surface tension. If not gold labeling, and these are bare gold colloids, you might be able to pull it off with either of the solvents. I don't have experience with trying. I don't know if you will have a clumping phenomenon from adding colloidal gold to polar solvents. You may want to add gold and ethanol together first and dry some down on a carbon-formvar grid and look at it in the TEM to see if the gold disperses evenly or not. A non-polar solvent such as Freon 113 is an alternative, dehydrating the gold by going from ethanol or acetone to the Freon, then to the sample. Again, I haven't tried the Freon. Another solvent to attempt would be HMDS, hexamethyldisilazane, again with ethanol or acetone as an intermediate step. You will probably have to sonicate in the Freon or HMDS if the gold has clumped in the ethanol or acetone. I don't have the colloidal gold to experiment with. I have IgG gold, but this will behave differently, having a different surface charge.

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: LSA 119
________________________________________
X-from: William.F.Tivol-at-aero.org [William.F.Tivol-at-aero.org]
Sent: Tuesday, December 14, 2010 12:39 PM
To: Haller, Edward

Dear Lists,
I have a specimen that is very small and not too firmly attached
to its substrate to which I would like to add colloidal gold. I am very
concerned that the surface tension in a few-ul drop of aqueous colloidal
gold will detach the specimen, so I would like to transfer the gold to a
solvent or suspension medium that has much lower surface tension, such as
ethanol or acetone. I am not concerned that either of these solvents
would harm the specimen, but they might not give a good dispersal of the
gold. Has anyone had experience with how gold disperses in various
solvents? TIA for any help.
Yours,
Bill

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From: William.F.Tivol-at-aero.org
Date: 12/14/2010 10:48 AM
Subject: [Microscopy] Other solvent for colloidal gold

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi Ed,
Thanks for your suggestions. I am not doing immunolabelling, just
bare gold colloids. I have no experience with HMDS, but freon could be
just the thing. I know that the growth of the colloids is stopped by
linking sulfur-containing organic chains to the Au atoms, so I don't know
whether the ethanol, acetone, or freon will mess things up, and clumping,
as you say, is an issue.
Yours,
Bill



X-from: "Haller, Edward" {ehaller-at-health.usf.edu}
To: "William.F.Tivol-at-aero.org" {William.F.Tivol-at-aero.org} ,
"microscopy-at-microscopy.com" {microscopy-at-microscopy.com}



Hi, Bill,

You didn't mention if you are doing immunolabeling or not, which would
change your approach to the application of the gold particles. If you are
immunolabeling, you might try increasing the concentration of Tween 20 in
your solution to lower surface tension. If not gold labeling, and these
are bare gold colloids, you might be able to pull it off with either of
the solvents. I don't have experience with trying. I don't know if you
will have a clumping phenomenon from adding colloidal gold to polar
solvents. You may want to add gold and ethanol together first and dry some
down on a carbon-formvar grid and look at it in the TEM to see if the gold
disperses evenly or not. A non-polar solvent such as Freon 113 is an
alternative, dehydrating the gold by going from ethanol or acetone to the
Freon, then to the sample. Again, I haven't tried the Freon. Another
solvent to attempt would be HMDS, hexamethyldisilazane, again with ethanol
or acetone as an intermediate step. You will probably have to sonicate in
the Freon or HMDS if the gold has clumped in the ethanol or acetone. I
don't have the colloidal gold to experiment with. I have IgG gold, but
this will behave differently, having a different surface charge.

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: LSA 119
________________________________________
X-from: William.F.Tivol-at-aero.org [William.F.Tivol-at-aero.org]
Sent: Tuesday, December 14, 2010 12:39 PM
To: Haller, Edward

----------------------------------------------------------------------------
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America

Dear Lists,
I have a specimen that is very small and not too firmly attached
to its substrate to which I would like to add colloidal gold. I am very
concerned that the surface tension in a few-ul drop of aqueous colloidal
gold will detach the specimen, so I would like to transfer the gold to a
solvent or suspension medium that has much lower surface tension, such as
ethanol or acetone. I am not concerned that either of these solvents
would harm the specimen, but they might not give a good dispersal of the
gold. Has anyone had experience with how gold disperses in various
solvents? TIA for any help.
Yours,
Bill

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From: larry.ackerman-at-ucsf.edu
Date: Tue, 14 Dec 2010 13:44:24 -0600
Subject: [Microscopy] Vessicle Suspension

Contents Retrieved from Microscopy Listserver Archives
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I have received a new (to me) type of specimen prep: 100nm vessicles in
suspension. They can be pelleted in an ultracentrifuge but due to the
small sample size encapsulation in agar or agarose seems risky--maybe 30
ul would work. Do you have any experience or suggestions for such a
sample prep (conventional glutaraldehyde/osmium/ethanol/epoxy)?

Thanks,
Larry
--
Larry Ackerman, Specialist
Electron Microscopy Lab
Manager, DERC Microscopy Core
UCSF, Dept. of Anatomy, Rm S1355
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

415-999-4758

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From: William.F.Tivol-at-aero.org
Date: 12/14/2010 11:54 AM
Subject: [Microscopy] Vessicle Suspension

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Dear Larry,
These vessicles are very well observed as cryo specimens. UCSF
has the equipment to plunge-freeze and examine them and to do tomography,
if that would give you useful information. Furthermore, the preparation
and microscopy are straightforward--in fact, this would be a good specimen
to introduce someone to cryoTEM. I would encourage you to ask people at
the Mission Bay facility about access to a cryo instrument.
Yours,
Bill



X-from: larry.ackerman-at-ucsf.edu
To: William.F.Tivol-at-aero.org



I have received a new (to me) type of specimen prep: 100nm vessicles in
suspension. They can be pelleted in an ultracentrifuge but due to the
small sample size encapsulation in agar or agarose seems risky--maybe 30
ul would work. Do you have any experience or suggestions for such a
sample prep (conventional glutaraldehyde/osmium/ethanol/epoxy)?


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From: dservando86-at-yahoo.com
Date: Tue, 14 Dec 2010 15:25:38 -0600
Subject: [Microscopy] viaWWW: cellular structure of decaying tissue

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Email: dservando86-at-yahoo.com
Name: Donya S.

Organization: SJ Delta College

Title-Subject: [Filtered] cellular structure of decaying tissue

Message: Hello, I am an EM student working on a project for my
advanced ultramicrotomy class. I chose to compare a freshly fixed
sample compared to a sample that underwent the process of decay for
three days before fixation. For the 3 day old liver tissue sample, I
could only find collagen fibers throughout my sample. Although,
there is another structure throughout my images and I am needing help
with identifying what it is. I was thinking it was fibroblast cells
or even elastic tissue, however, I could be completely off and it may
just be an artifact or contaminant of some sort???? Furthermore, I
have yet to be successful with my research in order to determine why
the collagen fibers are the only remaining structures during the
decay process. The only research I have found so far, would be the
increase of collagen fibers when the liver is induced with hepatic
fibrosis. Could someone PLEASE enlighten me with some direction for
my research?? I can email you my images if needed.

Thanks in advance for any advice!

Donya S.

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From: Lindsay.P.Keller-at-nasa.gov
Date: Tue, 14 Dec 2010 15:25:56 -0600
Subject: [Microscopy] viaWWW: PIPS conditions for sensitive samples

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Email: Lindsay.P.Keller-at-nasa.gov
Name: Lindsay Keller

Organization: NASA

Title-Subject: [Filtered] PIPS conditions for sensitive samples

Message: Does anyone have some recommended settings for a Gatan PIPS
to prepare TEM sections of geological samples containing clays and
carbonates? Thanks.

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From: hfong11-at-yahoo.com
Date: Tue, 14 Dec 2010 15:26:23 -0600
Subject: [Microscopy] viaWWW: Looking for a diffusion pump heater

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Email: hfong11-at-yahoo.com
Name: Hanson Fong

Organization: Univ of Washington

Title-Subject: [Filtered] Looking for a diffusion pump heater

Message: Does anyone have a heater for an Edwards 63PH2 diffusion
pump on hand and can sell to me right away? The Edwards part # is
H01700189 - they are currently out of stock.

Thanks,
Hanson

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From: parishcm-at-ornl.gov
Date: Tue, 14 Dec 2010 16:13:27 -0600
Subject: [Microscopy] TEM - tilting faulted materials

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Hello to all,

I'm working on X-ray analysis of grain boundaries in silicon carbide. The grain structure is highly columnar and full of faults. The problem I'm having is how to decide if my beam direction is straight down a boundary when there's so much diffraction contrast arising from faults and defects within each grain. For example, if I light up a grain in dark field, it's hard to tell if the inclined boundary is getting narrower as I tilt or if some sort of fault contrast is changing. I've done the obvious and oriented the columnar axis along my holder's alpha-tilt, but I'm not sure what the next step ought to be. Does anyone have any suggestions for how I might hit these grain boundaries?

Thanks
Chad Parish


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From: leunissen-at-aurion.nl
Date: Tue, 14 Dec 2010 18:12:07 -0600
Subject: [Microscopy] Re: Other solvent for colloidal gold

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Dear Bill,

If you wanted to exchange solutions with uncoated colloidal gold particles that will not likely be a successful enterprise and you will end up with serious aggregation. In my experience virtually all the colloid precipitates as the hydration layer on the gold surface collapses. The charge based repulsion which keeps the particles in solution will largely disappear with the hydration layer and merely London-vanderWaals forces remain which lead to an aggregation.
You might have some success with particles that are coated with a detergent-like molecule, as long as the coating substance is soluble both in water and in the solvent you would want to use and the particles are not to big. But this is just a (hopefully) educated guess.
There may be ways to produce gold particles in ethanol right from the start. Contact me off list if you are interested and I will try to find this.

Jan

Aurion
http://www.aurion.nl




On 15/12/2010, at 6:31 AM, William.F.Tivol-at-aero.org wrote:

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} Dear Lists,
} I have a specimen that is very small and not too firmly attached
} to its substrate to which I would like to add colloidal gold. I am very
} concerned that the surface tension in a few-ul drop of aqueous colloidal
} gold will detach the specimen, so I would like to transfer the gold to a
} solvent or suspension medium that has much lower surface tension, such as
} ethanol or acetone. I am not concerned that either of these solvents
} would harm the specimen, but they might not give a good dispersal of the
} gold. Has anyone had experience with how gold disperses in various
} solvents? TIA for any help.
} Yours,
} Bill



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From: donc-at-asmicro.com
Date: Tue, 14 Dec 2010 23:53:22 -0600
Subject: [Microscopy] Re: [a] worn tracball on S-4800 - your experiences?

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Bettina Wolpensinger asked about a trackball for an SEM.
Some AFMs, such as the Dimension 3100 in my lab, also use trackballs. The
specific type of trackball this AFM uses has 4 buttons and is seen by the PC
as a "serial mouse". Mechanical problems seem to relate to the bearings
that support the inner rods and/or accumulation of dirt on the rods. When
cleaning fails to solve the problem and the rods become noisy, then it is
time to replace the entire unit (not the ball only).

If your trackball has 4 buttons like mine and behaves as a serial mouse,
then possibly I can help you obtain a new unit.
Contact me offline - send photos of the front and back of your unit.
regards,
Don
=============================================
Don Chernoff, Ph.D., President
Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com
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----- Original Message -----
From: wolpensinger-at-isfh.de
To: donc-at-asmicro.com
Sent: Tuesday, December 14, 2010 2:05 AM
Subject: [a] [Microscopy] worn tracball on S-4800 - your experiences?





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Hi all,

After about 3 years the trackball-unit on our S-4800 is so heavily worn
that we now use it by removing the ball and turning the inner rods by hand.

When asking for a replacement, the company seemed to have a hard time, and
told us that the trackball unit does not have a part number, nor can it be
replaced solely.
Meanwhile we recieved a quote, but surprisingly the delivery is sheduled
in 90 days!! This gives me the impression that we might be the only users on
the planet being troubled with a worn trackball?!

Does anybody have similar experiences?
Any recommendations?
I've naiively wondered if it really needs a trackball, or if a simple
joystick could do the same job?

Thanks in advance,

Bettina





Bettina Wolpensinger
Institute for Solar Energy Research Hamelin (ISFH)
An-Institut der Leibniz Universitdt Hannover
Am Ohrberg 1
D-31860 Emmerthal
Germany
Tel: +49 5151 999 313
Fax: +49 5151 999 400
Email:
wolpensinger-at-isfh.de
Handelsregister: Amtsgericht Hannover HRB 100547
Geschdftsf|hrer: Prof. Dr. Rolf Brendel
Vorsitzender des Aufsichtsrats: Ministerialrat Dr. Hans Schroeder


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From: dcalvert-at-eastman.com
Date: Wed, 15 Dec 2010 07:47:58 -0600
Subject: [I] [Microscopy] viaWWW: Looking for a diffusion pump heater

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I got a heater from these folks http://www.daltonelectric.com/platen-heaters/

Looks and works just fine and they probably stock yours (and they are also probably cheaper than dealing with Edwards or your vendor)

Dave Calvert
Technical Associate
Eastman Chemical Co.
P.O. Box 1974
Kingsport, Tennessee 37662
Office: 423-229-4943
Cell: 423-963-8966
Fax: 423-224-7550

-----Original Message-----
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To: Calvert, Dave

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Email: hfong11-at-yahoo.com
Name: Hanson Fong

Organization: Univ of Washington

Title-Subject: [Filtered] Looking for a diffusion pump heater

Message: Does anyone have a heater for an Edwards 63PH2 diffusion
pump on hand and can sell to me right away? The Edwards part # is
H01700189 - they are currently out of stock.

Thanks,
Hanson

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From: edelmare-at-muohio.edu
Date: Wed, 15 Dec 2010 10:06:53 -0600
Subject: [Microscopy] Re: worn tracball on S-4800 - your experiences?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Well, trackballs and mice do wear out, as Don mentioned.

But they are still made and available for PC's and Mac's. Supporting both
USB, PS2, and even serial type connections. Trackballs and mice are fairly
simple systems and even without a driver specific for an older system I have
found them to be fairly interchangeable. The drivers do allow you to re-
program the four buttons. But the S-4800 runs under Windows doesn't it?
Then its not a problem.

A quick web search showed this site: http://www.trackballworld.com

It would seem like a pretty good place to start.


And www.newegg.com has sales on Kensington Trackballs (From
experience they are nice)

Good luck!


On 14 Dec 2010 at 2:03, wolpensinger-at-isfh.de wrote:

}
}
}
} ----------------------------------------------------------------------
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} of America To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver On-Line Help
} http://www.microscopy.com/MicroscopyListserver/FAQ.html
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}
} Hi all,
}
} After about 3 years the trackball-unit on our S-4800 is so heavily
} worn that we now use it by removing the ball and turning the inner
} rods by hand.
}
} When asking for a replacement, the company seemed to have a hard time,
} and told us that the trackball unit does not have a part number, nor
} can it be replaced solely. Meanwhile we recieved a quote, but
} surprisingly the delivery is sheduled in 90 days!! This gives me the
} impression that we might be the only users on the planet being
} troubled with a worn trackball?!
}
} Does anybody have similar experiences?
} Any recommendations?
} I've naiively wondered if it really needs a trackball, or if a simple
} joystick could do the same job?
}
} Thanks in advance,
}
} Bettina
}
}
}
}
}
} Bettina Wolpensinger
} Institute for Solar Energy Research Hamelin (ISFH)
} An-Institut der Leibniz Universität Hannover
} Am Ohrberg 1
} D-31860 Emmerthal
} Germany
} Tel: +49 5151 999 313
} Fax: +49 5151 999 400
} Email:
} wolpensinger-at-isfh.de
} Handelsregister: Amtsgericht Hannover HRB 100547
} Geschäftsführer: Prof. Dr. Rolf Brendel
} Vorsitzender des Aufsichtsrats: Ministerialrat Dr. Hans Schroeder
}
}
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Richard E. Edelmann, Ph.D.
Electron Microscopy Facility Director
364 Pearson Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-muohio.edu
http://www.emf.muohio.edu



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From: spurgeon-at-drexel.edu
Date: Wed, 15 Dec 2010 10:16:09 -0600
Subject: [Microscopy] Postdoctoral Research Position in Characterization of Irradiated

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dynamic Characterization Group

Department of Materials Science and Engineering - Drexel University

The Dynamic Characterization Group at Drexel University seeks to hire
a candidate for the position of Postdoctoral Research Associate. The
successful applicant will be primarily responsible for studying
microstructure-dependent radiation damage in metallic systems. The
individual will participate in grant writing, publication preparation,
and oral presentations as well as support additional projects within
the group.

A qualified applicant will possess a Ph.D. in materials science,
nuclear engineering, condensed matter physics, or a related field. The
desired candidate will be proficient in electron microscopy,
particularly TEM (e.g. HRTEM, STEM/EDS, EELS, in-situ) and SEM (e.g.
EBSD, FIB). Familiarity with radiation damage and experimental
radiation techniques, atom probe, and corrosion testing techniques is
beneficial. United States citizenship is preferred.

Interested applicants should  send a cover letter, curriculum vita,
and names of three references to Dr. Mitra Taheri
(mtaheri-at-coe.drexel.edu)

For more information, please visit our website at
http://dcg.materials.drexel.edu/

--
Steven Spurgeon
Dynamic Characterization Group
Department of Materials Science & Engineering
Drexel University

Office: Bossone 105
Email: spurgeon-at-drexel.edu
Web: http://www.materials.drexel.edu/dcg/


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From: edelmare-at-muohio.edu
Date: Wed, 15 Dec 2010 10:29:50 -0600
Subject: [Microscopy] Brands of plain glass slides

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

O.k., Gary:

Not sure if this part of the "OoO deluge" but since no one else has
asked so I will. Quartz? Your worried about the variations in coverslips
(Cracking? thicknesses?) and so you use Quartz?

Hmm, what about the Index of refraction? O.k., Fused Quartz Index of
refraction is 1.4585 (1.47 to 1.45). Which is really close to 1.473 Glycerol
(general mounintg medium particularly with the anti-fade agents). And it is
closer to water at 1.33 for wet mounts. Versus Glass at 1.515 hmm . . . .
actually quartz does sounds pretty good. Perhaps I'll give it a try, but now I
have to buy some 1.4585 oil.

As for the slides, I would have to agree that for Brightfield work I have used
both the green and the white sldies with little difference (I generally avoid the
ultra cheap really green slides, and go for the cleanest - none are really
clean but it makes it easier to clean them). And in general most folks do not
even think about lamp color temperature or accurately setting white balance
in the ditigal cameras.

For reflected light, epi-fluoresence or confocal work the slide color does not
matter.



On 12 Dec 2010 at 21:11, gary-at-gaugler.com wrote:

}
}
}
} ----------------------------------------------------------------------
} ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society
} of America To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver On-Line Help
} http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------
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}
} FWIW, I only use Corning 2947 LM slides. The big problem
} is finding good cover slips that are not made in China.
} These lousy units come pre-cracked. Wonderful. I pay more
} for quartz slips.
}
} Whatever the color transmittance is, I can correct it with
} digital capture software light balance.
}
} gary g.
}
} Here comes the OoO deluge.
}
}
} At 11:26 PM 12/11/2010, you wrote:
}
}
}
} } ---------------------------------------------------------------------
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} } http://www.microscopy.com/MicroscopyListserver/FAQ.html
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} }
} } I was wondering if anybody can explain what qualitative difference
} } exist, if any, between different brands of plain glass slides. For
} } example, over on the Ted Pella page (
} } http://www.tedpella.com/histo_html/slides.htm
} } ), for a 2-box pack of 144 slides, the price varies from about $10
} } for the Pella Economy Slides, all the way up to $40 or $60+ for Gold
} } Seal or Corning slides.
} }
} } My question is, is there any advantage or optical difference with the
} } more expensive slides? Corning advertises theirs as "water white"
} } glass, which I imagine implies something about the transmittance or
} } refractive index, but I'm not sure.
} }
} } ==============================Original
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Richard E. Edelmann, Ph.D.
Electron Microscopy Facility Director
364 Pearson Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-muohio.edu
http://www.emf.muohio.edu


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From: dservando86-at-yahoo.com
Date: Wed, 15 Dec 2010 12:01:14 -0600
Subject: [Microscopy] TEM- images displaying liver decay

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I want to thank all of those whom offered their advice in regards to the
decaying liver sample.  I have taken all the advice into consideration while
continuing my research; and all of you have given me a great head start!  For
those of you offering additional assistance by requesting to view my images, I
have tried to respond back, however, the emails were not successful due to a
"mailer-DAEMON" rejection.  So, are there other emails I may send them to?  I
would still very much appreciate your opinions on my images. 


Donya



----- Original Message ----
X-from: "Sobocinski, Gregg" {greggps-at-umich.edu}
To: "dservando86-at-yahoo.com" {dservando86-at-yahoo.com}
Sent: Tue, December 14, 2010 7:45:51 AM

Hello, I am an EM student working on a project for my advanced ultramicrotomy
class.  I chose to compare a freshly fixed sample compared to a sample that
underwent the process of decay for three days before fixation.  For the 3 day
old liver tissue sample, I could only find collagen fibers throughout my
sample.  Although, there is another structure throughout my images and I am
needing help with identifying what it is.  I was thinking it was fibroblast
cells or even elastic tissue, however, I could be completely off and it may just

be an artifact or contaminant of some sort????  Furthermore, I have yet to be
successful with my research in order to determine why the collagen fibers are
the only remaining structures during the decay process.  The only research I
have found so far, would be the increase of collagen fibers when the liver is
induced with hepatic fibrosis.  Could someone PLEASE enlighten me with some
direction for my research?? I can email you my images if needed.
Thanks in advance for any advice!
Donya S.


     


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From: gary-at-gaugler.com
Date: Wed, 15 Dec 2010 12:21:04 -0600
Subject: [Microscopy] Re: Brands of plain glass slides

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Here are German glass slides and covers:

http://www.knittel-glaeser.de/index_e.htm

I have used these slides and especially the
covers. Very high quality. There used to be
a US distributor but not sure any more. I
bought 500 slides and 500 covers and have not
run out yet. The Corning slides are easy to
get and are clear. Standard 1x3".

gary g.

At 11:26 PM 12/11/2010, you wrote:



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From: b-myers3-at-northwestern.edu
Date: Wed, 15 Dec 2010 12:50:14 -0600
Subject: [Microscopy] Re: worn tracball on S-4800 - your experiences?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I had 5 different stages of decay, in addition to a freshly fixed sample for
comparison.  There are 3 hrs, 12 hrs, 24 hrs, and 52 hrs.  The problem I ran
into was that we didn't do a 48 hr stage.  So, there is a vast difference in
cellular decay from 24 hrs to 52 hrs.  Thus it is making it rather difficult to
further understand the rate of decay.  All of the samples had been placed into
closed petri dishes at approximately room temperature until put into 4F:1G
fixative.  I had replied back to the actual respondants, and the emails were
still rejected.  And I didn't even think about posting them into an online
database; good idea!



----- Original Message ----
X-from: Derrick Horne {dhorne-at-interchange.ubc.ca}
To: dservando86-at-yahoo.com
Sent: Wed, December 15, 2010 10:11:42 AM

Hi Bettina-

Our S-4800 and S-3400 have had similar issues as they get a LOT of use
around here. Anyhow, our service engineer was able to adjust the
position of the "rods" in the track ball for better contact and it works
pretty well now. I'm not exactly sure how to do this myself, but if you
contact me directly I can put you or your service engineer in contact
with our engineer. I don't think this system is compatible with a
standard trackball mouse, but I could be wrong.

Regards,
Ben

On 12/14/2010 1:13 AM, wolpensinger-at-isfh.de wrote:
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} Hi all,
}
} After about 3 years the trackball-unit on our S-4800 is so heavily worn that we now use it by removing the ball and turning the inner rods by hand.
}
} When asking for a replacement, the company seemed to have a hard time, and told us that the trackball unit does not have a part number, nor can it be replaced solely.
} Meanwhile we recieved a quote, but surprisingly the delivery is sheduled in 90 days!! This gives me the impression that we might be the only users on the planet being troubled with a worn trackball?!
}
} Does anybody have similar experiences?
} Any recommendations?
} I've naiively wondered if it really needs a trackball, or if a simple joystick could do the same job?
}
} Thanks in advance,
}
} Bettina
}
}
}
}
}
} Bettina Wolpensinger
} Institute for Solar Energy Research Hamelin (ISFH)
} An-Institut der Leibniz Universität Hannover
} Am Ohrberg 1
} D-31860 Emmerthal
} Germany
} Tel: +49 5151 999 313
} Fax: +49 5151 999 400
} Email:
} wolpensinger-at-isfh.de
} Handelsregister: Amtsgericht Hannover HRB 100547
} Geschäftsführer: Prof. Dr. Rolf Brendel
} Vorsitzender des Aufsichtsrats: Ministerialrat Dr. Hans Schroeder
}
}
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}

--
*Ben Myers*
SEM/FIB Facility Manager
NU/ANCE/ Center

Northwestern University
Mail: 2036 Cook Hall
Office: 1114 Cook Hall
2220 Campus Drive
Evanston, IL 60208-3108

ph: (847) 491-3439
fax: (847) 467-6573

http://www.nuance.northwestern.edu

==============================Original Headers==============================
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From: gstrout-at-ou.edu
Date: Wed, 15 Dec 2010 17:57:05 -0600
Subject: [Microscopy] D500i dimpler problem

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

We have been experiencing a recurring problem with our D500i dimpler.
When the arm down button is pushed, the arm arm does not slow down and
make gentle contact, but instead slams into the specimen platen. In
addition, the F/Z toggle switch does not work. The machine is stuck in F
set mode, which has no effect.
The machine has been sent out for repair for similar problems several
times now only to be returned and have the same problem after working
for just a short period of time. We have received no information from
the repair service about the nature of the problem and would like to see
if there are any suggestions from the listserve before we give up and
purchase a new dimpler.

Greg Strout
Research Scientist
Samuel Roberts Noble Microscopy Laboratory
University of Oklahoma
770 Van Vleet Oval
Norman, Ok, 73019

eMail: gstrout-at-ou.edu
Phone: 405-325-4391




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From: dhorne-at-interchange.ubc.ca
Date: Wed, 15 Dec 2010 18:57:24 -0600
Subject: [Microscopy] viaWWW: Zinc Oxide film and SAED

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: dhorne-at-interchange.ubc.ca
Name: Derrick Horne

Organization: UBC BioImaging Facility

Title-Subject: [Filtered] Zinc Oxide film and SAED

Message: We have a user trying to characterize zinc oxide films
dip-cast and calcined (450C) onto a glass slide. Apparently there
should be diffraction differences through the thickness of the film
from the air side to the glass. Preserving the spatial orientation is
imperative.

My original recommendation to our user was ion milling and/or FIB to
create a thin enough sample for TEM. This equipment is not readily
available to her or to us at this time.

I tried peeling off the layers using tape, but I was unable to pull
off the complete film from air to glass. That which did come off the
slide bonded poorly to the tape and epoxy resin, and while
sectionable to some extent, the orientation of the film was lost.

I'm not expecting any silver bullets, but perhaps some guidance from
those of you in the materials world might nudge us in a different
direction.

Thanks in advance.



Login Host: 137.82.85.211
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From: swalck-at-southbaytech.com
Date: Wed, 15 Dec 2010 19:42:08 -0600
Subject: [Microscopy] viaWWW: Zinc Oxide film and SAED

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Derrick,

You can prepare cross sections of ZnO coated samples using the
MicroCleave(TM) technique, (aka Small Angle Cleavage Technique). Glass is a
material that can be used.

I do know that some of the thin films of ZnO that I have examined had an
initial equiaxed grain area that transitioned to a columnar grain structure
and that some of the samples showed a discontinuous fracture along that
transition zone. There still were areas to be seen and the microstructure
could be evaluated. The MicroCleave(TM) technique is a very inexpensive
method of preparing great samples for materials that it works for. It does
work for glass samples.

Please go to our application notes section of our website,
http://southbaytech.com/applist.htm, and open app note #62. While there,
you might also want to look at #55, #59, #60, #61 also.

I see that you are from Canada. John McCaffrey, your fellow countryman,
originally developed the technique.

Disclaimer: SBT manufactures and sells the MicroCleave(TM) kit.

-Scott

Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA 92673

eMail: SWalck-at-SouthBayTech.com
Phone: 949-492-2600
Toll Free: 1-800-728-2233 (USA)
Fax: 949-492-1499

www.SouthBayTech.com



-----Original Message-----
X-from: dhorne-at-interchange.ubc.ca [mailto:dhorne-at-interchange.ubc.ca]
Sent: Wednesday, December 15, 2010 5:04 PM
To: swalck-at-southbaytech.com

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
---------------------------------------------------------------------------
Remember this posting is most likely not from a Subscriber, so when replying
please copy both dhorne-at-interchange.ubc.ca as well as the
MIcroscopy Listserver
---------------------------------------------------------------------------

Email: dhorne-at-interchange.ubc.ca
Name: Derrick Horne

Organization: UBC BioImaging Facility

Title-Subject: [Filtered] Zinc Oxide film and SAED

Message: We have a user trying to characterize zinc oxide films
dip-cast and calcined (450C) onto a glass slide. Apparently there
should be diffraction differences through the thickness of the film
from the air side to the glass. Preserving the spatial orientation is
imperative.

My original recommendation to our user was ion milling and/or FIB to
create a thin enough sample for TEM. This equipment is not readily
available to her or to us at this time.

I tried peeling off the layers using tape, but I was unable to pull
off the complete film from air to glass. That which did come off the
slide bonded poorly to the tape and epoxy resin, and while
sectionable to some extent, the orientation of the film was lost.

I'm not expecting any silver bullets, but perhaps some guidance from
those of you in the materials world might nudge us in a different
direction.

Thanks in advance.



Login Host: 137.82.85.211
---------------------------------------------------------------------------

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==============================Original Headers==============================
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From: jsb43-at-cam.ac.uk
Date: Thu, 16 Dec 2010 05:07:21 -0600
Subject: [Microscopy] Re: Zinc Oxide film and SAED

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Derrick,

You could try removing the glass by back-thining with a dilute solution of
hydrofluoric acid. You need to coat your sample with a thin layer of
HF-resistant material (e.g. lacomit) but leave a small window over the
glass that you want to remove:

1. Cut a small piece of the sample and mount on a metal slot/hole grid with
the ZnO in contect with the metal- an epoxy-based glue, like Araldite, will
establish a conduction path to discharge the sample during observation.

2. Mount the sample onto a plastic dipper (or something that will resist HF
attack) using superglue or some other removeable adhesive.

3. Use a liquid like Lacomit to coat the areas of the sample that are not
to be etched, but leave a small hole ( {1 mm diameter) over the centre of
the glass that needs to the removed.

4. Either dip the stick into an HF solution or drip the HF-solution using a
plastic dropper. Please do a risk assessment *before* using this!

5. Thoroughly rinse the sample with fresh de-ionised water before placing
under an optical microscope to monitor the etch rate. You should see Newton
fringes appear when the glass is getting very thin (~micrometres). You will
probably find that the HF attacks the ZnO too, but with a little care you
could probably get just the glass removed leaving just enough ZnO to be
observed.

6. Once finished, rinse with lacomit remover and adhesive remover to clean
the sample. Wash in methanol and ethanol and leave to dry. The sample
should be ready for study.

7. If you find the sample charges, coat with a thin layer of carbon (a few
nanometers should do). It will give some amorphous contrast in the
diffraction pattern, but not much.

Please note that HF is extremely dangerous to handle, so access to it might
be difficult to obtain. Please consult Materials Data Safety Sheets (MSDS)
or COSSH forms (UK-based chemical safety standard) to do a proper risk
assessment.

Good luck.

Jon

} Email: dhorne-at-interchange.ubc.ca
} Name: Derrick Horne
} Title-Subject: [Filtered] Zinc Oxide film and SAED
}
} Message: We have a user trying to characterize zinc oxide films
} dip-cast and calcined (450C) onto a glass slide. Apparently there
} should be diffraction differences through the thickness of the film
} from the air side to the glass. Preserving the spatial orientation is
} imperative.
}
} My original recommendation to our user was ion milling and/or FIB to
} create a thin enough sample for TEM. This equipment is not readily
} available to her or to us at this time.
}
} I tried peeling off the layers using tape, but I was unable to pull
} off the complete film from air to glass. That which did come off the
} slide bonded poorly to the tape and epoxy resin, and while
} sectionable to some extent, the orientation of the film was lost.
}
} I'm not expecting any silver bullets, but perhaps some guidance from
} those of you in the materials world might nudge us in a different
} direction.
}
} Thanks in advance.

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From: mullarkay-at-gmail.com
Date: Thu, 16 Dec 2010 14:13:24 -0600
Subject: [Microscopy] Aberration-corrected TEM Position

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The College of Nanoscale Science and Engineering at the University at
Albany, part of the SUNY system, is looking to hire a research
scientist in the area of aberration-corrected transmission electron
microscopy (TEM). Duties will include, but not be limited to: managing
the FEI Titan TEM facility, and working closely with the CNSE faculty,
staff and strategic partners to develop new methodologies for
nanomaterials analysis and semiconductor metrology. Effort will be
split between advanced applications support and advanced technique
development.

For more information and how to apply please see:

http://cnse.albany.edu/Careers/Details.aspx?career_id=5bc6fbf0-d91c-4057-b852-3312acc0b58a


Thanks,

Tom Murray

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6, 30 -- Subject: Aberration-corrected TEM Position
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From: pakhomov-at-uw.edu
Date: Thu, 16 Dec 2010 17:05:58 -0600
Subject: [Microscopy] viaWWW: Job at UW (Seattle, WA) NanoTech User Facility

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Email: pakhomov-at-uw.edu
Name: Alec Pakhomov

Organization: U of Washington

Title-Subject: [Filtered] Job at UW (Seattle, WA) NanoTech User Facility

Message: Hi everyone,

The NanoTech User Facility (NTUF, https://depts.washington.edu/ntuf)
at the University of Washington (Seattle, WA) has an outstanding
opportunity for a RESEARCH SCIENTIST 2 ( TEM SPECIALIST) .

The job details, requirements and application method can be found at:

https://uwhires.admin.washington.edu/ENG/candidates/default.cfm?szLocationID=88&szCategory=Jobprofile&szOrderID=69755&szCandidateID=0&szSearchWords=&szReturnToSearch=1

UW HR Req #: 69755
Open until filled

For informal inquiries, please contact me by e-mail:
pakhomov-at-uw.edu

Alec Pakhomov
Lab Manager/Principal Research Scientist
NanoTech User Facility
Center for Nanotechnology
Fluke Hall 215
Box 352140
University of Washington
Seattle, WA 98195-2140

e-mail: pakhomov-at-uw.edu
https://depts.washington.edu/ntuf


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From: dhorne-at-interchange.ubc.ca
Date: Thu, 16 Dec 2010 17:06:54 -0600
Subject: [Microscopy] viaWWW: Combustion of polymerized Epon 812- ZnO question continued

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Email: dhorne-at-interchange.ubc.ca
Name: Derrick Horne

Organization: UBC BioImaging Facility

Title-Subject: [Filtered] Combustion of polymerized Epon 812- ZnO
question continued

Message: I know this is going to come across as an odd question, but
in an attempt to avoid the use of HF acid in the prepping of the
dip-cast slides for electron diffraction of zinc oxides, we came up
with the idea of dip casting on Epon instead of glass. We thought
this would facilitate sectioning of the film.

Does anyone know the temperature at which polymerized Epon burns?

I've done some Google searching, but haven't come up with anything definitive.

Thanks in advance,

Derrick

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From: gstrout-at-ou.edu
Date: Thu, 16 Dec 2010 18:40:26 -0600
Subject: [Microscopy] Old VCR Group dimpler problem

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Thanks to everyone who responded to my question about the arm problem on
our VCR Group dimpler. I was looking for information about repairs we
might be able to make to the machine ourselves and was in no way making
comments about previous service. If that is how it was taken I
apologize as this was not the intent of my request. Nearly all the
responses were about minor maintenance and lubrication being the only
real issues they'd had with their dimplers. Our dimpler is 18 years old
and has been in twice in the last couple of years for repair which
considering its age is not a bad track record.
South Bay Technology responded to me very quickly and we are currently
talking with them about repairs to the machine. Many thanks for the
responses from the folks on the microscopy listserve and thanks again to
South Bay Technology for their quick and informative response.

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From: rfoley-at-uab.edu
Date: Thu, 16 Dec 2010 21:11:15 -0600
Subject: [Microscopy] viaWWW: Biological SEM users - what do you need and what do you

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Email: rfoley-at-uab.edu
Name: Robin Foley

Organization: UAB

Title-Subject: [Filtered] Biological SEM users - what do you need
and what do you desire?

Message: All,

I'm a materials SEM user but will be getting involved more with
biological-type specimens in the future. We've been shopping for a
new SEM and I'm hoping to get some opinions about what SEM features
and add-ons that biological SEM users think are absolutely necessary.
Also, if you had an endless budget - what features would you spring
for?

Thanks,
Robin Foley, UAB

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From: gary-at-gaugler.com
Date: Thu, 16 Dec 2010 21:40:59 -0600
Subject: [Microscopy] Re: viaWWW: Biological SEM users - what do you

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

That is rather wide open but interesting.

If biological ONLY, then the maker does not
make any or much difference. The high resolution SEMs
will have magnetic immersion final lenses and
will not tolerate Ferrous specimens. If you ever
anticipate imaging Ferrous specimens at high
resolution, the only option is Zeiss Gemini
column...totally electrostatic.

CFE versus TFE is a separate issue. But if no
long scans or maps, then mox nix. For pure
high rez images, the Hitachi CFE is IMO the choice.
But you have the task of flashing the tip each day.
Plus side is that the tips are much cheaper than
the Schottky TFE guns. But the TFEs are more stable.
For short pix captures, it should not make any difference.

Go dry pumping everywhere. Dry scroll roughing pump(s)
and dual mag-lev turbo. Do not get DP. A dry specimen
interchange lock is also a must. Dry and fast specimen
exchange.

Scintillator BSE like Robinson over solid state.
Then add STEM with solid state BF/DF multi quadrant
capability.

SDD EDS with triple monitors--two for SEM and one
for EDS. Multiple ways to adjust probe current
to optimize cps versus DT. But need high cps and low DT
to do quick EDS maps. This can be done.

Are you more interested in imaging or elemental analysis
or both? Hitachi will probably do better in imaging.
Regardless, be prepared for bone crushing maintenance cost.
Down time is also an issue. All tools suffer from this.
If they are treated well, that helps. One stupid act
can trash the system and cost big time...dollars and time.

Give more details about what you want to do and perhaps
I can supply more input. You will be fixing and preparing
the specimens accordingly, right? What about a coater?

gary g.

At 07:12 PM 12/16/2010, you wrote:
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From: jeff-at-quorumtechnologies.com
Date: Fri, 17 Dec 2010 08:25:15 -0600
Subject: [Microscopy] viaWWW: SEM: Quorum Technologies and Energy Beam (Commercial

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Email: jeff-at-quorumtechnologies.com
Name: Jeff Butler

Organization: Quorum

Title-Subject: [Filtered] SEM: Quorum Technologies and Energy Beam
(Commercial Response)

Message: Hi All:

We wanted send a general note to make a distinction between the two
Quorum Technologies currently operating in the microscopy market in
hopes that users are not frustrated if they are trying to contact
either company.

Quorum Technologies, based in Guelph Canada is a supplier focussed on
Light microscopy and the components required for Live Cell Imaging
and can be found at www.quorumtechnologies.com.

Quorum Technologies based in the United Kingdom is focused on
Electron Microscopy and can be found online at
http://www.quorumtech.com/.

Sorry for the commercial post, but we've encountered some frustrated
microscopists trying to get information from the wrong company.

Best Regards,
Jeff Butler

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From: sharonl-at-uwindsor.ca
Date: Fri, 17 Dec 2010 09:56:31 -0600
Subject: [Microscopy] viaWWW: SEM image stitching

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Email: sharonl-at-uwindsor.ca
Name: Sharon Lackie

Organization: University of Windsor

Title-Subject: [Filtered] SEM image stitching

Message: Can anyone suggest good software to stitch multiple SEM
images together to make one large image from multiple frames? The
software I've tried all assumes they are landscape images, and
accounts for angle changes, or a curved lens, and I end up with a
curved image. Thanks.

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From: fclaabs-at-iastate.edu
Date: Fri, 17 Dec 2010 12:31:29 -0600
Subject: [Microscopy] Etec Auto Scan available

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I have an Etec Auto Scan looking for a new home. In it's day this was the very top of the line SEM. It would make and excellect item for a sci fi movie prop. It has more knobs and screens than one could imagine, sort of like a Star Trek set. If you want it you must move it. I would prefer it go to some non profit like a school but that is not necessary (I'll give it to them). If no one wants it it will end up as scrap because I need to reclaim the space.
Fran Laabs
Ames IA 50014

515 232 7933

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From: hyi-at-emory.edu
Date: Fri, 17 Dec 2010 13:49:55 -0600
Subject: [Microscopy] Anti-GFAP antibody

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Dear All:

Can anyone recommend a monoclonal anti-GFAP antibody that has been
proven to work well on thin sections of embedded brain tissue. Thank you in
advance.

Happy holidays.

Hong


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From: maryflet-at-interchange.ubc.ca
Date: Fri, 17 Dec 2010 15:19:59 -0600
Subject: [Microscopy] viaWWW: SEM image stitching

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Dear Sharon,
Type "autostitch" into Google and you will get the program developed at UBC
that will do just that. I don't think it will distort the images and it is
free for Windows.

Mary Fletcher
Electron Microscopist
Materials Eng. UBC
#309 - 6350 Stores Road
Vancouver, B.C. V6T 1Z4
Canada
Tel: 604-822-5648
Fax: 604-822-3619
email: maryflet-at-interchange.ubc.ca


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Email: sharonl-at-uwindsor.ca
Name: Sharon Lackie

Organization: University of Windsor

Title-Subject: [Filtered] SEM image stitching

Message: Can anyone suggest good software to stitch multiple SEM
images together to make one large image from multiple frames? The
software I've tried all assumes they are landscape images, and
accounts for angle changes, or a curved lens, and I end up with a
curved image. Thanks.

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From: Jerzy.Gazda-at-ceriumlabs.com
Date: Fri, 17 Dec 2010 15:40:35 -0600
Subject: [Microscopy] viaWWW: SEM image stitching

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Sharon,

Depending on the manufacturer of the SEM, the software might be available from them, double check with applications support. I have used a commercial software (not free) called SIGMAGIS from SmartImaging Technology. The company can assist you with writing scripts or automating the process if you have large data files. My image arrays range from 100x100 to much larger. The software will compensate for lens distortions etc...

For small jobs, you can investigate either Google or Microsoft products for Gigapixel imaging downloadable from the web, also ImageJ and Digital Micrograph has scripts that can stitch few images together.


Happy Holidays,

Jerzy
PS: I have no financial interest in software packages, just a happy customer.

***************************************************
Jerzy Gazda Ph.D.
Section Manager - TEM, FIB, SEM
Cerium Laboratories LLC
5204 E. Ben White Blvd. MS-512
Austin, TX 78741

(512) 934-5185 vm
(512) 622-6600 pgr

www.ceriumlabs.com


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Name: Sharon Lackie

Organization: University of Windsor

Title-Subject: [Filtered] SEM image stitching

Message: Can anyone suggest good software to stitch multiple SEM
images together to make one large image from multiple frames? The
software I've tried all assumes they are landscape images, and
accounts for angle changes, or a curved lens, and I end up with a
curved image. Thanks.

Login Host: 137.207.232.55
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From: kwylly-at-ppg.com
Date: Fri, 17 Dec 2010 17:28:58 -0600
Subject: [Microscopy] viaWWW: Cross-Sectional Polishers

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Email: kwylly-at-ppg.com
Name: Kelly Wylly

Organization: PPG Industries

Title-Subject: [Filtered] Cross-Sectional Polishers

Message: I have been investigating cross-sectional polishers from
JEOL, Hitachi and Gatan and am interested in any feedback from users
that currently own this equipment.

The instruments that are being evaluated are the JEOL IB-09010 CP,
Gatan's Ion Beam Cutter and Hitachi's E-3500.

Has anyone used these polishers as preparation for AFM analysis or
metallurgical experiments using EBSD?

How successful are these polishers at site-specific cross-sections?

I would also be interested in finding local owners of these
instruments in order to more fully explore their utitlity for my
samples. I am located in the Pittsburgh area.

Do any contract labs offer this type of sample preparation as a service?

Any help or feedback would be greatly appreciated.

Kelly Wylly
Group Leader - Microscopy Lab
PPG - Coatings Innovation Center
412-492-5625



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From: gcanzalo-at-mtu.edu
Date: Fri, 17 Dec 2010 17:29:42 -0600
Subject: [Microscopy] viaWWW: Automated acquisition of mosaic/tile SE images

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Email: gcanzalo-at-mtu.edu
Name: Jerry Anzalone

Organization: Michigan Technological University

Title-Subject: [Filtered] Automated acquisition of mosaic/tile SE images

Message: Good day:

I'm trying to automate acquisition of a series of images over a
sample that I can then either perform PSD image analysis on and/or
tile to make a large, high resolution "panorama", if you will. Using
a FEI XL40 ESEM and we've got an EDAX workstation running v3.3 of EDX
Imaging/Mapping and v3.3 of EDX Particle/Phase Analysis with column
and stage control. To date, I've been unable to get either
application to store only an image at the stage coordinates stored in
the stage table - they want to perform their designed function and
collect spectra. I don't want to wait until the next election cycle
to get my data...

Can anybody tell me if it is possible to do what I want with the
software on hand or point out another solution?

Thank you for your time.

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From: allan.mitchell-at-stonebow.otago.ac.nz
Date: Fri, 17 Dec 2010 18:59:36 -0600
Subject: [Microscopy] Re: Etec Auto Scan available

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi all

We also have recently be faced with the problem of disposing of a couple of older EM's to make may for new technology coming in. I am always very reluctant to see such wonderful technology and engineering go to the scrap yard or dump. However, we have found a place to dispose of older equipment , if no other home can be found. Although the equipment will end up being modified to a large extent, at least it finds a new use.

A town 100 k's north of Dunedin called Oamaru, is building up a large following in steam punk. See web links below.

There are all manner of fittings, pipe work, consoles, etc that these guys love. A recent exhibition held in Oamaru included a Dr Gattling's Lunar Dismembulator. This device inlcudes HT components and gun from an old seimens 102 TEM that we once had.

So if you can't find a home for your old EM's, but don't want to see them go to the tip, find your local steam punk enthusiasts group and offer it to them.

Anyone finding themselves passing through Oamaru in a 12 months time, check out the steam punk submarine control room at the Steam Punk exhibition hall.

http://steampunkoamaru.co.nz/

Have great christmas period everyone

Allan



On 18/12/2010, at 7:39 AM, {fclaabs-at-iastate.edu} wrote:

}
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} I have an Etec Auto Scan looking for a new home. In it's day this was the very top of the line SEM. It would make and excellect item for a sci fi movie prop. It has more knobs and screens than one could imagine, sort of like a Star Trek set. If you want it you must move it. I would prefer it go to some non profit like a school but that is not necessary (I'll give it to them). If no one wants it it will end up as scrap because I need to reclaim the space.
} Fran Laabs



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From: Barrie.Wells-at-ConwyValley.com
Date: Sat, 18 Dec 2010 02:34:32 -0600
Subject: [Microscopy] Re: viaWWW: SEM image stitching

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Thank you, Mary, but we do sometimes see distortion in our stitched images
when we use "autostitch": not always, but enough times to cause us problems.
Our images are from petrographic thin sections, i.e. transmitted light
microscopy, if that might make a difference.
Are we alone in seeing this? If so, we must consider that it is our usage,
not the software itself, so I would appreciate replies either way.

Thank you,

Barrie Wells
Conwy Valley Systems Limited

}
} Dear Sharon,
} Type "autostitch" into Google and you will get the program developed at
} UBC
} that will do just that. I don't think it will distort the images and it is
} free for Windows.
}
} Mary Fletcher
} Electron Microscopist
} Materials Eng. UBC
} }
} }
} } Title-Subject: [Filtered] SEM image stitching
} }
} } Message: Can anyone suggest good software to stitch multiple SEM
} } images together to make one large image from multiple frames? The
} } software I've tried all assumes they are landscape images, and
} } accounts for angle changes, or a curved lens, and I end up with a
} } curved image. Thanks.
} }



-----
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Internal Virus Database is out of date.


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From: parishcm-at-ornl.gov
Date: Sat, 18 Dec 2010 09:44:23 -0600
Subject: [Microscopy] RE: viaWWW: Cross-Sectional Polishers

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Kelly,

At my previous institution, we had a JEOL cross-sectioner that I used to successfully prepare easily-oxidized thin films on silicon substrates for cross-sectional EBSD (See: Journal of Nuclear Materials, V403, P. 191-197, 2010). I also used it to cross-section thick films of ceramic on a glassy substrate and obtained excellent SEM-EDS-EBSD results.

I am unfamiliar with the Gatan system so I can't comment on it positively or negatively, but I would love to have either the JEOL or the Hitachi in my lab. I found this concept to be an excellent way to make cross-sections for SEM-EDS-EBSD. In principal, it would also be a good way to prepare cross-sections to pull FIB liftouts from, but I haven't actually tried that.

I think that "site specific," however, will be limited to +/- a few hundred microns, but check with the applications / sales engineers on that.

Chad Parish


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From: swalck-at-southbaytech.com
Date: Sat, 18 Dec 2010 23:21:27 -0600
Subject: [Microscopy] Re: viaWWW: Cross-Sectional Polishers

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Kelly,
I would also like to sugest that you look at the South Bay Technology IBS/e that is equipped with the KDC-10 ion source. This ion source is a Kaufman type and is capable of treating an area of up to about 1-3/4 inches in diameter. It is a low energy, high current, large area beam. It has been sucessfully used for preparing EBSD sample at low angle and low energy (200 eV) and very good results have been presented at the M&M meetings. Since it is a Kaufman ion source, it can be used with a neutralizer and the beam can be completely neutralized. This works well for ion polishing or ion etching non-conductive samples or samples that have been mounted in metallurgical mounts. A new slope cutting sample holder is available that can slope cut samples at an angle of 0°, 45°, and 90° with a 1" wide mask. Recent experiments have shown sputter rates (with focusing optics) can conservatively be about 30 times that of a conventional high energy, low current, focusing ion gun system. For metallurgical samples, the system can be used to ion polish and then ion etch samples to enhance optical and scanning electron microscopy images.

Disclaimer: South Bay Technology manufactures and sells the IBS/e system. We have an exclusive arrangement with Kaufman and Robinson Incorporated to incorporate their ion source and power supply with our system.


-Scott

Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA 92673

US Toll Free: 1-800-728-2233
Tel: (949) 492-2600
Fax: (949) 492-1499

www.southbaytech.com
SWalck-at-SouthBayTech.com




On Fri 17/12/10 3:37 PM , kwylly-at-ppg.com sent:
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}
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} http://microscopy.com/MicroscopyListserver/MLFormMail.html---------------------------------------------------------------------------
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} replyingplease copy both kwylly-at-ppg.com
} as well as the MIcroscopy Listserver---------------------------------------------------------------------------
}
} Email: kwylly-at-ppg.com
} Name: Kelly Wylly
}
} Organization: PPG Industries
}
} Title-Subject: [Filtered] Cross-Sectional Polishers
}
} Message: I have been investigating cross-sectional polishers from
} JEOL, Hitachi and Gatan and am interested in any feedback from users
} that currently own this equipment.
}
} The instruments that are being evaluated are the JEOL IB-09010 CP,
} Gatan's Ion Beam Cutter and Hitachi's E-3500.
}
} Has anyone used these polishers as preparation for AFM analysis or
} metallurgical experiments using EBSD?
}
} How successful are these polishers at site-specific cross-sections?
}
} I would also be interested in finding local owners of these
} instruments in order to more fully explore their utitlity for my
} samples. I am located in the Pittsburgh area.
}
} Do any contract labs offer this type of sample preparation as a
} service?
} Any help or feedback would be greatly appreciated.
}
} Kelly Wylly
} Group Leader - Microscopy Lab
} PPG - Coatings Innovation Center
} 412-492-5625
}
}
}
} Login Host: 141.189.251.1
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From: gary-at-gaugler.com
Date: Sat, 18 Dec 2010 23:59:57 -0600
Subject: [Microscopy] Re: viaWWW: Cross-Sectional Polishers

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

This seems on first blush as a sledge hammer for an ant.
I might be wrong.

Why not use an Ultramet polishing unit with coloidal silica
and/or diamond slurry? Either will give good results
for EBSD. Just watch your film thickness and ensure that is is even enough
to be sure that you have an area that is polished and
can be scanned.

gary g.


At 03:30 PM 12/17/2010, you wrote:



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From: swalck-at-southbaytech.com
Date: Sun, 19 Dec 2010 01:55:55 -0600
Subject: [Microscopy] viaWWW: Cross-Sectional Polishers

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Gary,

Colloidal silica polishing for some difficult to polish samples does't give the best results for EBSD.

With low energy ion polishing at low angle (250 eV at 4°), we have taken Ti-6Al-4V samples that have been polished for 2 min and 30 minutes by colloidal silica and compared them. The 30 minutes samples have better hit rates and band contrast histogram distributions than do the 2 minutes by a significant amount. However, after ion polihsing these samples, they give consistent and much improved samples. In another test, ion polishing aftermechanical polishing the samples beat samples that were vibratory polished with colloidal silica for extended times.

Ion polishing and slope cutting can do samples that are too soft or too delecated for mechanical polishing. In addition, cross sectional polishing by mechanical means such as tripod polishing does some training and experience to get good results. The slope cutting technique can be quite easily done by anyone with very little training.



-Scott

Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA 92673

US Toll Free: 1-800-728-2233
Tel: (949) 492-2600
Fax: (949) 492-1499

www.southbaytech.com
SWalck-at-SouthBayTech.com




On Sat 18/12/10 10:09 PM , gary-at-gaugler.com sent:
}
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}
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} -
} This seems on first blush as a sledge hammer for an ant.
} I might be wrong.
}
} Why not use an Ultramet polishing unit with coloidal silica
} and/or diamond slurry? Either will give good results
} for EBSD. Just watch your film thickness and ensure that is is even
} enoughto be sure that you have an area that is polished and
} can be scanned.
}
} gary g.
}
}
} At 03:30 PM 12/17/2010, you wrote:
}
}
}
} } -------------------------------------------------------
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} } Email: kwylly-at-ppg.com
} } Name: Kelly Wylly
} }
} } Organization: PPG Industries
} }
} } Title-Subject: [Filtered] Cross-Sectional
} Polishers}
} } Message: I have been investigating
} cross-sectional polishers from} JEOL, Hitachi and Gatan and am interested in any
} feedback from users} that currently own this equipment.
} }
} } The instruments that are being evaluated are the
} JEOL IB-09010 CP,} Gatan's Ion Beam Cutter and Hitachi's
} E-3500.}
} } Has anyone used these polishers as preparation
} for AFM analysis or} metallurgical experiments using EBSD?
} }
} } How successful are these polishers at
} site-specific cross-sections?}
} } I would also be interested in finding local
} owners of these} instruments in order to more fully explore their
} utitlity for my} samples. I am located in the Pittsburgh
} area.}
} } Do any contract labs offer this type of sample
} preparation as a service?}
} } Any help or feedback would be greatly
} appreciated.}
} } Kelly Wylly
} } Group Leader - Microscopy Lab
} } PPG - Coatings Innovation Center
} } 412-492-5625
} }
} }
} }
} } Login Host: 141.189.251.1
} } -------------------------------------------------------
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} } ==============================Original
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} (by way of MicroscopyListserver)} 15, 22 -- Subject: viaWWW: Cross-Sectional
} Polishers} 15, 22 -- Content-Type: text/plain;
} charset="us-ascii" ; format="flowed"} ==============================End of -
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==============================Original Headers==============================
15, 20 -- From swalck-at-southbaytech.com Sun Dec 19 01:55:54 2010
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From: parishcm-at-ornl.gov
Date: Sun, 19 Dec 2010 08:00:01 -0600
Subject: [Microscopy] Re: viaWWW: Cross-Sectional Polishers

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Gary,

You make a good point. However, in my mind, the two main advantages of an ion cross sectioner relative to mechanical polishing are:

1. Dry vacuum. The samples I was dealing with would have been destroyed by any sort of polishing solution because they oxidized so easily. (I had to run them from the ion polisher to the SEM to get patterns at all).

2. Labor. Even very thick specimens take only a few minutes of work (labor charges) -- ten minutes to load, polish overnight, five minutes to unload.

In my humble opinion, only certain subsets problems are appropriate for the ion cross-sectioners, but those problems really require nothing but.

Chad Parish
________________________________________
X-from: gary-at-gaugler.com [gary-at-gaugler.com]
Sent: Sunday, December 19, 2010 1:10 AM
To: Parish, Chad M.

This seems on first blush as a sledge hammer for an ant.
I might be wrong.

Why not use an Ultramet polishing unit with coloidal silica
and/or diamond slurry? Either will give good results
for EBSD. Just watch your film thickness and ensure that is is even enough
to be sure that you have an area that is polished and
can be scanned.

gary g.




==============================Original Headers==============================
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From: PhillipsT-at-missouri.edu
Date: Sun, 19 Dec 2010 10:28:17 -0600
Subject: [Microscopy] Re: viaWWW: SEM image stitching

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Photoshop allows one to select whether stitched images are corrected for perspective or cylindrical layouts or just be repositioned without correction. It is the Photomerge function under Automate.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: Barrie.Wells-at-ConwyValley.com [mailto:Barrie.Wells-at-ConwyValley.com]
Sent: Saturday, December 18, 2010 2:35 AM
To: Phillips, Thomas E.

Thank you, Mary, but we do sometimes see distortion in our stitched images
when we use "autostitch": not always, but enough times to cause us problems.
Our images are from petrographic thin sections, i.e. transmitted light
microscopy, if that might make a difference.
Are we alone in seeing this? If so, we must consider that it is our usage,
not the software itself, so I would appreciate replies either way.

Thank you,

Barrie Wells
Conwy Valley Systems Limited

}
} Dear Sharon,
} Type "autostitch" into Google and you will get the program developed at
} UBC
} that will do just that. I don't think it will distort the images and it is
} free for Windows.
}
} Mary Fletcher
} Electron Microscopist
} Materials Eng. UBC
} }
} }
} } Title-Subject: [Filtered] SEM image stitching
} }
} } Message: Can anyone suggest good software to stitch multiple SEM
} } images together to make one large image from multiple frames? The
} } software I've tried all assumes they are landscape images, and
} } accounts for angle changes, or a curved lens, and I end up with a
} } curved image. Thanks.
} }



-----
No virus found in this message.
Checked by AVG - www.avg.com
Version: 10.0.1152 / Virus Database: 424/3209 - Release Date: 10/20/10
Internal Virus Database is out of date.


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From: vakimler-at-med.wayne.edu
Date: Sun, 19 Dec 2010 14:00:03 -0600
Subject: [Microscopy] viaWWW: Antigen Unmasking

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Antigen Unmasking

Message: I would like to try one of the techniques of Stirling and
Graff (1995)J. Histochem Cytochem, to unmask a challenging antigen
that I am labeling with enhanced fluoronanogold.

If sodium metaperiodate is not available in time for the experiment,
would periodic acid work, too? If yes, how would that be prepared?
The protocol calls for incubation of tissue sections on grids in a
humidified chamber with a saturated solution of sodium
metaperiodate. Tissues will then be treated with the blocking
solution that has been successful.

Thanks!

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From: tomw-at-uidaho.edu
Date: Sun, 19 Dec 2010 14:00:33 -0600
Subject: [Microscopy] viaWWW: Bruker D8 XRD Request

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Email: tomw-at-uidaho.edu
Name: Tom Williams

Organization: University of Idaho

Title-Subject: [Filtered] Bruker D8 XRD Request

Message: A friend working at Dublin City University is having issues
with their XRD and needs access to the schematics for the 2-Theta
drive board for a Bruker D8 XRD. Obtaining it from the vendor is,
not unexpectedly, very costly. If anyone can assist, I can pass
along contact information, or put you in touch.

Thanks,
Tom


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From: kraftpiano-at-gmail.com
Date: Sun, 19 Dec 2010 14:21:41 -0600
Subject: [Microscopy] EDS LN2 boiling off rapidly.

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I've got an Oxford EDS detector that all of a sudden just can't hold its LN2. I filled it last night, and today it is empty and the outside of the dewar is sweating like crazy. I know there's a good vacuum in the detector itself, which leads me to believe that the dewar is somehow bad. It was working fine two weeks ago, when it could easily go two to three days without a top off. Nothing has happened since to the detector- does anybody know what the issue could be? I can't imagine that the dewar itself somehow went bad for no reason...

--Justin A. Kraft

"America believes in education; the average professor earns more money in a year than a professional athlete earns in a whole week." Evan Esar

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From: kenconverse-at-qualityimages.biz
Date: Sun, 19 Dec 2010 16:20:11 -0600
Subject: [Microscopy] Etec Auto Scan available

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Fran (et al),
They're still a great instrument if your needs aren't real extravagant. I
consider them to be the most "student proof" SEM out there. As they say,
"Nothing is foolproof because fools are so damned ingenious."

If it moves to the northeast, I can provide full support.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


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X-from: fclaabs-at-iastate.edu [mailto:fclaabs-at-iastate.edu]
Sent: Friday, December 17, 2010 1:34 PM
To: kenconverse-at-qualityimages.biz

I have an Etec Auto Scan looking for a new home. In it's day this was the
very top of the line SEM. It would make and excellect item for a sci fi
movie prop. It has more knobs and screens than one could imagine, sort of
like a Star Trek set. If you want it you must move it. I would prefer it go
to some non profit like a school but that is not necessary (I'll give it to
them). If no one wants it it will end up as scrap because I need to reclaim
the space.
Fran Laabs
Ames IA 50014

515 232 7933

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From: marshall-at-sfu.ca
Date: Sun, 19 Dec 2010 22:30:07 -0600
Subject: [Microscopy] EDS LN2 boiling off rapidly

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Hello:
There are a oouple of reasons for N2 boiling off quickly. Often it is a poor vacuum in the dewar walls. Sometimes it is ice or some other foreign substance in the bottom of the dewar. In your case I suspect the vacuum even though you say it is fine. The sweating generally means no insulation (i.e. no vacuum). The other interesting thing is that your N2 only lasted 2 or 3 days. In a normal 8 liter dewar with good vacuum, it should last 9 or 10 days. If you have a smaller dewar then the time may be much shorter, but if your dewar is the standard 7 or 8 liters then I suspect it has had a bad vaccum for a while and has finally failed. Could you please let me know off-site some more information about the capacity of your dewar, but I really am suspicious of your vacuum within the dewar. How do you know it is good?
Dan

Dan Marshall
Professor of Geochemistry and Economic Geology
Simon Fraser University
Burnaby, BC, V5A 1S6
CANADA

marshall-at-sfu.ca www.sfu.ca/~marshall

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From: kraftpiano-at-gmail.com
Date: Sun, 19 Dec 2010 22:48:59 -0600
Subject: [Microscopy] Re: EDS LN2 boiling off rapidly- clarification.

Contents Retrieved from Microscopy Listserver Archives
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I realized, based on the replies, that I wasn't specific enough in my description of what I've done so far. I don't think it's a window issue because the detector is one with a thin window, a Be window, and a no window position on a selector. I tested the vacuum by opening the window, pumping the chamber down as best I could, then shutting the whole thing down while a vacuum gauge monitored it overnight. The next morning, there was no vacuum deterioration in the chamber, and therefore no vacuum deterioration that could be detected in the detector itself. It has been suggested that perhaps the vacuum in the dewar and the vacuum in the detector are two separate isolated vacuum chambers- I don't know. I'll check on that. As to repairs, I have no budget for repair. I do have an older detector that I believe to be functional that is compatible with the analysis machine (An old Link eXL) which I can use as a replacement of this one, I just have to change out the fitting that interfaces between the block end of the detector and the wall of the chamber. I was trying to avoid using the old detector because the thin window is gone, but the Be window is still there. The older detector is a Link LZ-4, and the new detector (That has the issue) is a Pentafet detector.

At this point, I think the dewar being bad is probably the most likely scenario. I half-filled it with LN2 this morning and let it sit, and the first part to become "Sweaty" was the top of the dewar around the fill hole.

Incidentally, does anyone have access to software to use the data files from an old eXL on a PC? I have the PC-Link setup working, but I can't do anything with the files after I've transferred them.

Since this is going towards educational use, (Read: I have no budget except what's in my wallet) I was actually thinking of replacing it with a solid state device from a company I found called AmpTek (www.amptek.com). They produce very reasonably priced detectors, around $2900 for their entry level model, but I'm not sure what software I would be able to use with it, or how I would link it up to do elemental mapping. The main features I'm looking for are to be able to do elemental mapping and basic analysis. Has anyone had experience with these detectors? Feel free to contact me offline with comments both good and bad.

--Justin.

On Dec 19, 2010, at 3:25 PM, kraftpiano-at-gmail.com wrote:

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} I've got an Oxford EDS detector that all of a sudden just can't hold its LN2. I filled it last night, and today it is empty and the outside of the dewar is sweating like crazy. I know there's a good vacuum in the detector itself, which leads me to believe that the dewar is somehow bad. It was working fine two weeks ago, when it could easily go two to three days without a top off. Nothing has happened since to the detector- does anybody know what the issue could be? I can't imagine that the dewar itself somehow went bad for no reason...
}
} --Justin A. Kraft
}
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From: wolpensinger-at-isfh.de
Date: Mon, 20 Dec 2010 01:48:58 -0600
Subject: [Microscopy] worn trackball - THANKS!!

Contents Retrieved from Microscopy Listserver Archives
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Thank you so much for all your responses to my trackball enquiry!

Your shared experiences and hints make me feel confident that we will solve the problem rather sooner than later and most likely without spending much money.

In order to work on the trackball I only need to manage to get to my workplace... We've got such an unusual amount of snow (fresh and older) here in northern Germany, that I gave up waiting for the early morning train and I'm back home now. Maybe tomorrow will be better!

Regards
Bettina



Bettina Wolpensinger
Institut für Solarenergieforschung Hameln/Emmerthal GmbH (ISFH)
An-Institut der Leibniz Universität Hannover
Am Ohrberg 1
D-31860 Emmerthal
Germany
Tel: +49 5151 999 313
Fax: +49 5151 999 400
Email: wolpensinger-at-isfh.de

Handelsregister: Amtsgericht Hannover HRB 100547
Geschäftsführer: Prof. Dr. Rolf Brendel
Vorsitzender des Aufsichtsrats: Ministerialrat Dr. Hans Schroeder





Bettina Wolpensinger
Institut für Solarenergieforschung Hameln/Emmerthal GmbH (ISFH)
An-Institut der Leibniz Universität Hannover
Am Ohrberg 1
D-31860 Emmerthal
Germany
Tel: +49 5151 999 313
Fax: +49 5151 999 400
Email:
wolpensinger-at-isfh.de
Handelsregister: Amtsgericht Hannover HRB 100547
Geschäftsführer: Prof. Dr. Rolf Brendel
Vorsitzender des Aufsichtsrats: Ministerialrat Dr. Hans Schroeder


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From: jacques.faerber-at-ipcms.u-strasbg.fr
Date: Mon, 20 Dec 2010 02:42:10 -0600
Subject: [Microscopy] Re: viaWWW: SEM image stitching

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi

I've had the same problem, as most "panorama" softwares are built to be
used with images coming from a camera, they suppose one is in a conique
projection system.

There are a few plugins for ImageJ (MosaicJ, Large montage, Patchwork)
which are more or less manual. I tested only "Large Montage".

I got some good results with Hugin, a open software using the Panotools
library (I use it on linux, but it exist too for win and OSX). As it
calculate the focal length, it jams on the result it gets, as the value
is much too low for what is normaly wait ! Focal length of a 0.5 to 3
mm, field of view from 100 µm to a few mm are not common values for
panorama makers ! On must force it to accepte these values, and it work
further.

With SEM images, I had always to set the control points manually, as the
B&W makes a bit more complicate the automatic location process. It
depends of the type of picture you have.
The projection type is set to rectilinear.
One can either give an initial field of view value (or a focal lenght),
or let it calculate them itself, and fine tune the result. In that case,
in the menu "Optimisation", one choose the option "all".
At the end of the computation, one can again modify more or less the
focal lenght, to minimize the distortions of the montage.
I had nice results with sets of up to 20-30 images, I didn't try with
more images.

Hope it helps


J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr


On 17/12/2010 17:00, sharonl-at-uwindsor.ca wrote:
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} Email: sharonl-at-uwindsor.ca
} Name: Sharon Lackie
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} Organization: University of Windsor
}
} Title-Subject: [Filtered] SEM image stitching
}
} Message: Can anyone suggest good software to stitch multiple SEM
} images together to make one large image from multiple frames? The
} software I've tried all assumes they are landscape images, and
} accounts for angle changes, or a curved lens, and I end up with a
} curved image. Thanks.
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} ==============================Original Headers==============================
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From: W.Muss-at-salk.at
Date: Mon, 20 Dec 2010 06:34:02 -0600
Subject: [Microscopy] Re: Antigen Unmasking

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Vickie,
(Dear all),

since I have not seen a reply to this question on the Listserver up to now, I'm trying to provide some hopefully correct hints:

first of all:
I always was confident/thought, that 'sodium metaperiodate (HIO4)' and 'periodic acid (H5IO6)' were "the same" substance (depending on how the powder was brought into solution with water) but now I was puzzled about that specific request and considered to } pimp my brain { with looking for answers to that special question....

http://environmentalchemistry.com/yogi/chemicals/cn/Periodic%A0acid,%A0sodium%A0salt.html

•Synonyms/Related:
â—¦Metaperiodate
â—¦Periodic acid (HIO4), sodium salt
â—¦Periodic acid, sodium salt
â—¦Sodium m-periodate
â—¦Sodium metaperiodate
â—¦Sodium periodate

(from: http://www.patentstorm.us/patents/4082743/description.html )
Aqueous solution of meta periodate ions:
The oxidizing agent thus may have as its source a meta periodate of sufficient water solubility to be useful, and free from metallic ions which may adversely affect the course of the reaction.
Sodium meta periodate is a preferred example of such a compound. Periodic acid, derived by electrolytic processes from iodic acid, or by chemical oxidation of elemental iodine, also may be employed. In the latter case adjustment of the pH of the periodic acid product to a value of pH 2 to pH 4.6 is required. ........
Where sodium meta periodate is employed as the source of the oxidizing meta periodate ions, establishment of a pH of greater than 4.6 results in the conversion of the water soluble meta periodate to the water insoluble para periodate in accordance with the following equation:

NaIO4 2 NaOH → Na3 H2 IO6 ↓

from: http://cshprotocols.cshlp.org/cgi/content/full/2008/6/pdb.prot5016?print=true
(Cite as: Cold Spring Harb Protoc; 2008; doi:10.1101/pdb.prot5016
...... on that web-page some 2 pages below the initial header :....... "In this method, the osmium tetroxide is removed from the superficial layers of the section by treatment with periodic acid and/or sodium metaperiodate (Skepper et al. 1998). Thin sections are floated on drops of the oxidizing agent of choice, e.g., 5% (w/v) sodium metaperiodate for 10-20 min, and then rinsed thoroughly with ultrapure H2O before commencing immunochemical staining. Periodic acid and sodium metaperiodate are both oxidizing agents with differing efficacies. Some workers use only sodium metaperiodate, while others suggest that a sequential treatment with both produces stronger immunochemical staining. We tend to use a single treatment with sodium metaperiodate, which removes osmium tetroxide from the surface of the thin section, and in some cases this will enhance the binding of an antibody to its antigen at that surface.
(whole website/protocol dealing with Immunostaining - testing new antibodies - Fixation - Antigen preservation - getting rid of OsO4, - etc.


Also I would like to point you to an interesting article on this "staining matter"
(pdf with free online access -at- http://jhc.sagepub.com/content/50/1/11.full.pdf+html )
by MVT Lobo et al in JHC 2002:
Ultrastructural Staining with Sodium Metaperiodate_Na-Periodate(NaIO4)&Sodium(Na-)Borohydride.
(in their lit. ref's also is mentioned: STIRLING & GRAFF 1995)


So, IMHO, it should be possible to use also periodic acid with some differing efficiacy compared to metaperiodate (see above in "CSH Protocols...").

Hope this my post being a starting point for a fruitful discussion from/with colleagues dealing with such techniques every day...


Apologize, if this post got too "long",


Season's greetings
Merry Christmas and a Happy, healthy, successful and prosperous 2011 to you all and yours,

regards

Wolfgang MUSS
EM-LAb Pathology Gen. Hosp.
SALZBURG
Austria



} -----Ursprüngliche Nachricht-----
} Von: vakimler-at-med.wayne.edu [mailto:vakimler-at-med.wayne.edu]
} Gesendet: Sonntag, 19. Dezember 2010 21:05
} An: Muß Wolfgang
} Betreff: [Microscopy] Antigen Unmasking
[TEM-Ultrathin sections on grid: sodium metaperiodate vs periodic acid? Technique acc.to STIRLING
} & GRAFF (1995) in JHC]

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} Email: vakimler-at-med.wayne.edu
} Name: Vickie Kimler
} Organization: Wayne State University School of Medicine
} Title-Subject: Antigen Unmasking
} Message:
}
} I would like to try one of the techniques of Stirling and Graff (1995)J. Histochem Cytochem, to unmask a
} challenging antigen that I am labeling with enhanced fluoronanogold.
}
} If sodium metaperiodate is not available in time for the experiment, would periodic acid work, too?
}
} If yes, how would that be prepared?
} The protocol calls for incubation of tissue sections on grids in a humidified chamber with a saturated solution of sodium metaperiodate.
} Tissues will then be treated with the blocking solution that has been successful.
}
} Thanks!
}
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From: nizets2-at-yahoo.com
Date: Mon, 20 Dec 2010 07:35:07 -0600
Subject: [Microscopy] looking for distributors for Austria

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I hope I am not braking a rule here with this Email because it is directly
addressed to private companies but they are also part of the EM community I
think.
We are not happy with our current austrian (NOT australian!!) provider of
consumables for EM and we would like to consider others.
I would be grateful if the major and minor US companies providing consumables
for EM would care to send us their distributors for Austria or Germany. Feedback
from german/austrian customers would also be appreciated.
Thank you very much in advance.

Regards,

Stephane





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From: maryflet-at-interchange.ubc.ca
Date: Mon, 20 Dec 2010 11:07:34 -0600
Subject: [Microscopy] Re: viaWWW: SEM image stitching

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Barrie and Sharon and all,
I'm sorry, I should have made it clear that I do not use the Autostitch
program, just that I have heard other members of the Listserver mention it
and it is free.
Regards,

Mary Fletcher
Electron Microscopist
Materials Eng. UBC
#309 - 6350 Stores Road
Vancouver, B.C. V6T 1Z4
Canada
Tel: 604-822-5648
Fax: 604-822-3619
email: maryflet-at-interchange.ubc.ca


-----Original Message-----
X-from: Barrie.Wells-at-ConwyValley.com [mailto:Barrie.Wells-at-ConwyValley.com]
Sent: December 18, 2010 12:43 AM
To: maryflet-at-interchange.ubc.ca

Thank you, Mary, but we do sometimes see distortion in our stitched images
when we use "autostitch": not always, but enough times to cause us problems.
Our images are from petrographic thin sections, i.e. transmitted light
microscopy, if that might make a difference.
Are we alone in seeing this? If so, we must consider that it is our usage,
not the software itself, so I would appreciate replies either way.

Thank you,

Barrie Wells
Conwy Valley Systems Limited

}
} Dear Sharon,
} Type "autostitch" into Google and you will get the program developed at
} UBC
} that will do just that. I don't think it will distort the images and it is
} free for Windows.
}
} Mary Fletcher
} Electron Microscopist
} Materials Eng. UBC
} }
} }
} } Title-Subject: [Filtered] SEM image stitching
} }
} } Message: Can anyone suggest good software to stitch multiple SEM
} } images together to make one large image from multiple frames? The
} } software I've tried all assumes they are landscape images, and
} } accounts for angle changes, or a curved lens, and I end up with a
} } curved image. Thanks.
} }



-----
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Checked by AVG - www.avg.com
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From: sergei2-at-ornl.gov
Date: Mon, 20 Dec 2010 16:37:17 -0600
Subject: [Microscopy] ISAF-PFM-2011, Joint Conference, Vancouver

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Media Cybernetics, Image Pro Plus software can stitch images together.
This software is written for images from many different sources, whether
they are taken from an optical microscope, electron microscope, X-rays, or
ultrasound, etc.. I have never seen a distortion at the edges when
stitching the image back together. You need to have some overlap between
each image, so the software can see where to line up landmarks though. We
use this software all the time.

Cheryl Rehfeld
Meyer Instruments, Inc.
Phone 225-769-4465
Cell 225-281-7739
csr-at-meyerinst.com
----- Original Message -----
X-from: {maryflet-at-interchange.ubc.ca}
To: {csr-at-meyerinst.com}
Sent: Monday, December 20, 2010 11:09 AM

Dear Colleagues

We cordially invite you and your colleagues to participate in /The 20th
IEEE International Symposium on Applications of Ferroelectrics/ and /The
International Symposium on Piezoresponse Force Microscopy & Nanoscale
Phenomena in Polar Materials/, ISAF-PFM-2011, which will jointly take
place in Vancouver, British Columbia, Canada, from July 24 to 27, 2011.
http://www.sfu.ca/isaf-2011/ or http://www.sfu.ca/pfm-2011/

ISAF-2011 is part of a series of international conferences administrated
by the Ultrasonics, Ferroelectrics and Frequency Control (UFFC) Society
of the Institute of Electric and Electronic Engineers (IEEE). It is
aimed to provide a forum to present and discuss the state-of-the-art
developments in the field of ferroelectrics and related materials and
their applications, including theory and modeling, materials preparation
and characterization and device physics and processing. A special
session will be devoted to relaxor-based high-performance piezoelectric
single crystals.

PFM-2011 will feature tutorials on the principles and applications of
PFM to ferroelectric and multiferroic materials, as well as new areas of
PFM applied to strongly correlated oxides, energy storage and conversion
materials, point defects and nanoionics. The recent advances in imaging
of conventional ferroelectrics and polar materials as well as time and
voltage spectroscopy of static and dynamic domain structures will be
presented.

The joint ISAF-PFM-2011 Conference will feature the following *Plenary
Speakers*:
• *Akira Ando* (Murata Manufacturing Company, Japan)
• *Stuart Foster* (University of Toronto, Canada)
• *Steve Pennycook* (Oak Ridge National Lab., USA)
• *Ramamoorthy Ramesh* (University of California, Berkeley, USA)
• *Alexander Tagantsev* (Ecole Polytechnique Fédérale, Switzerland)
• *Zhong Lin Wang* (Georgia Institute of Technology, USA)
• *Dragan Damjanovic* (EPFL, Switzerland, /IEEE Distinguished Lecturer/)

Joint *ISAF-PFM Short Courses* will be offered by the following tutors
on Sunday, July 24, 2011:
• *Alexei Gruverman* (University of Nebraska-Lincoln)
• *Sergei V. Kalinin *(Oak Ridge National Laboratory)
• *Petro Maksymovytch* (Oak Ridge National Laboratory)
• *Paul Reynolds* (Weidlinger Associates, Inc.)
• *Susan Trolier-McKinstry* (Penn State University)

*Poster Competitions* will be organized and *Poster Awards *will be
presented to the winning students. Limited financial support to student
participants is available upon request.

Please note the following *important dates*:
• *Deadline for Abstract Submission: Mar. 01, 2011*
• Acceptance Notice: April 15, 2011
• Early Registration: May 01 - June 30, 2011
• Accommodation Reservation: May 01 - June 30, 2011
• Short Courses: July 24, 2011
• Conference Dates: July 24 - 27, 2011
• Deadline of Proceedings Manuscript Submission: July 25, 2011

More detailed information can be found on the attached brochure and on
the conference website at
http://www.sfu.ca/isaf-2011/ or http://www.sfu.ca/pfm-2011/

We look forward to welcoming you in the beautiful City of Vancouver in
July 2011.

Sincerely,

Zuo-Guang Ye Andrei Kholkin
General Chair General Chair
Simon Fraser University University of Aveiro
Canada Portugal

Thomas R. Shrout Sergei Kalinin
Technical Chair Technical Chair
Penn State University Oak Ridge National Lab.
USA USA

Contact Information:
Zuo-Guang Ye; Jessica Wong
Email: {isaf-pfm-2011-at-sfu.ca}

==============================Original Headers==============================
14, 25 -- From sergei2-at-ornl.gov Mon Dec 20 16:37:16 2010
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From: bill.mcmanus-at-usu.edu
Date: Mon, 20 Dec 2010 17:11:36 -0600
Subject: [Microscopy] Changing oil in Pfeiffer-Balzer 170 THP series turbo pump

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Does anyone know the protocol for changing oil in the Pfeiffer-Balzer
170 THP series turbo pump used on the Zeiss 902 TEM? I have the TL
011 oil and I know that each side gets 20 mls, but do you pour it in,
force it in or is there a trick. It just seems to not to what to
come out or go back in easily.

Bill McManus
Research Scientist
Western Dairy Center
Utah State University
bill.mcmanus-at-usu.edu


==============================Original Headers==============================
3, 20 -- From prvs=963e2d890=bill.mcmanus-at-usu.edu Mon Dec 20 17:11:36 2010
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From: bill.mcmanus-at-usu.edu
Date: Mon, 20 Dec 2010 17:14:27 -0600
Subject: [Microscopy] changing oil in the Pfeiffer-Balzer 170 THP turbo pump

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Does anyone know the protocol for changing oil in the Pfeiffer-Balzer
170 THP series turbo pump used on the Zeiss 902 TEM? I have the TL
011 oil and I know that each side gets 20 mls, but do you pour it in,
force it in or is there a trick. It just seems to not to what to
come out or go back in easily.

Bill McManus
Research Scientist
Western Dairy Center
Utah State University
bill.mcmanus-at-usu.edu


==============================Original Headers==============================
3, 20 -- From prvs=963e2d890=bill.mcmanus-at-usu.edu Mon Dec 20 17:14:27 2010
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From: kenconverse-at-qualityimages.biz
Date: Mon, 20 Dec 2010 18:28:47 -0600
Subject: [Microscopy] changing oil in the Pfeiffer-Balzer 170 THP turbo pump

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Bill,
I was working on a DSM950 SEM that had a 170 and called Pfeiffer. It seems
a little counter-intuitive, but they told me to just pull the pump out, lay
it on its side with the plugs out of both sides of both bearings and pour
the oil through. Most of it just flows out (flushing the inards) and what
remains is what you need.

I still don't feel real comfortable with that procedure, but it was straight
from the Pfeiffer service department. Who am I to argue?

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: bill.mcmanus-at-usu.edu [mailto:bill.mcmanus-at-usu.edu]
Sent: Monday, December 20, 2010 6:16 PM
To: kenconverse-at-qualityimages.biz

Does anyone know the protocol for changing oil in the Pfeiffer-Balzer
170 THP series turbo pump used on the Zeiss 902 TEM? I have the TL
011 oil and I know that each side gets 20 mls, but do you pour it in,
force it in or is there a trick. It just seems to not to what to
come out or go back in easily.

Bill McManus
Research Scientist
Western Dairy Center
Utah State University
bill.mcmanus-at-usu.edu


==============================Original Headers==============================
3, 20 -- From prvs=963e2d890=bill.mcmanus-at-usu.edu Mon Dec 20 17:14:27 2010
3, 20 -- Received: from ironport1.usu.edu (ironport1.usu.edu [129.123.1.26])
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delsp=yes
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3, 20 -- Subject: changing oil in the Pfeiffer-Balzer 170 THP turbo pump
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==============================Original Headers==============================
16, 28 -- From kenconverse-at-qualityimages.biz Mon Dec 20 18:28:46 2010
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16, 28 -- To: {bill.mcmanus-at-usu.edu} , "MSA Listserver" {microscopy-at-microscopy.com}
16, 28 -- Subject: RE: [Microscopy] changing oil in the Pfeiffer-Balzer 170 THP turbo pump
16, 28 -- Date: Mon, 20 Dec 2010 19:28:30 -0500
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From: kenconverse-at-qualityimages.biz
Date: Mon, 20 Dec 2010 21:15:11 -0600
Subject: [Microscopy] changing oil in the Pfeiffer-Balzer 170 THP turbo pump

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Kerry,
This is not the horizontal "dual" pump (TPU 330). It's a vertical one, but
I was told to just lay it on its side and run the oil through it. I agree
with your tips on the 330.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: Kerry Gascoigne [mailto:kerry.gascoigne-at-flinders.edu.au]
Sent: Monday, December 20, 2010 9:52 PM
To: kenconverse-at-qualityimages.biz

Bill,
I was working on a DSM950 SEM that had a 170 and called Pfeiffer. It seems
a little counter-intuitive, but they told me to just pull the pump out, lay
it on its side with the plugs out of both sides of both bearings and pour
the oil through. Most of it just flows out (flushing the inards) and what
remains is what you need.

I still don't feel real comfortable with that procedure, but it was straight
from the Pfeiffer service department. Who am I to argue?

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: bill.mcmanus-at-usu.edu [mailto:bill.mcmanus-at-usu.edu]
Sent: Monday, December 20, 2010 6:16 PM
To: kenconverse-at-qualityimages.biz

Does anyone know the protocol for changing oil in the Pfeiffer-Balzer
170 THP series turbo pump used on the Zeiss 902 TEM? I have the TL
011 oil and I know that each side gets 20 mls, but do you pour it in,
force it in or is there a trick. It just seems to not to what to
come out or go back in easily.

Bill McManus
Research Scientist
Western Dairy Center
Utah State University
bill.mcmanus-at-usu.edu


==============================Original Headers==============================
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==============================Original Headers==============================
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{microscopy-at-microscopy.com}
16, 28 -- Subject: RE: [Microscopy] changing oil in the Pfeiffer-Balzer 170
THP turbo pump
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==============================Original Headers==============================
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31, 29 -- Subject: RE: [Microscopy] RE: changing oil in the Pfeiffer-Balzer 170 THP turbo pump
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From: gary-at-gaugler.com
Date: Tue, 21 Dec 2010 00:19:40 -0600
Subject: [Microscopy] Re: viaWWW: SEM image stitching

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

COMMERCIAL REPLY:
Smart Imaging Technologies has stitched over 19,000 SEM images together into
a single image, about 1 billion pixels. We have the ability to stitch
multi-gigabit files. If customization is required, we may require some
sample images to provide cost for stitching.

We do offer a free file sharing and stitching service (for select
individuals and academia) that has fewer features than commercial version.
We can help you avoid the crescent moon phenomenon that you are
experiencing.

We also offer ready-to-run image analysis solutions as an online service
based on monthly subscription.

With SIMAGIS LiveT Online Virtual Microscopy you can:

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overlays; automatically tag and measure target objects, download
measurements, markers (objects of interest) and data reports. (commercial
version only)

For more information, please contact me offline:

Ira Bleiweiss
ira-at-simagis.us
Smart Imaging Technologies
713.589.3216 direct
877.280.1100 ext.1004 Toll Free (US)


www.smartimtech.com
Advanced Software Solutions for Science and Industry

-----Original Message-----
X-from: sharonl-at-uwindsor.ca [mailto:sharonl-at-uwindsor.ca]
Sent: Friday, December 17, 2010 10:03 AM
To: ira-at-simagis.us

You could try Zeiss or Raith and get their SmartStitch
app. It is dedicated to SEM stitching. There are other
apps that do stitching but this one is quite good.

gary g.


At 07:57 AM 12/17/2010, you wrote:



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From: jacques.faerber-at-ipcms.u-strasbg.fr
Date: Tue, 21 Dec 2010 02:24:48 -0600
Subject: [Microscopy] Re: changing oil in the Pfeiffer-Balzer 170 THP turbo

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi

I confirm what Ken told.
Put the pump (unmouted from the SEM) horizontally on two wedges, unscrew
the paires of bright slit screws mounted in opposite, one in the middle
of the pump, the other at the motor side. I have a funnel sold by
Pfeiffer which is to be scewed on place of the upper screw, with a mark
at the oil level. It's usful to avoid oil leaks at the funnel, as the
holes are smale, but one can do it with a very smale funnel, or a little
tube conected to a funnel. Poor the quantity of TL11 oil in the funnel
and it flow slowly trough the wick which is around the ball bearing. No
pressure at the inlet, only gravity !
Keep care at the beginning to let loose some bubbles which can block the
flow at the inlet with a piece of piano wire.
Put a smale dish under the lower screw hole, to collect the dirty oil.
You'll see the oil coming out first very durty, and becomming more and
more clean, until you have collected a little less then what you have
poored in.
It takes some time (1/4 h for each bearing), as you must wait until it
has almost finished to drop.

Same thing for both ball bearings, and it's good for 1 year (in
continuous use) .

By the way, the old oil is very fine for your bicycle chain, or other
such uses !

Hope it helps

J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr


On 21/12/2010 00:18, bill.mcmanus-at-usu.edu wrote:
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} Does anyone know the protocol for changing oil in the Pfeiffer-Balzer
} 170 THP series turbo pump used on the Zeiss 902 TEM? I have the TL
} 011 oil and I know that each side gets 20 mls, but do you pour it in,
} force it in or is there a trick. It just seems to not to what to
} come out or go back in easily.
}
} Bill McManus
} Research Scientist
} Western Dairy Center
} Utah State University
} bill.mcmanus-at-usu.edu
}
}
} ==============================Original Headers==============================
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From: peter.heimann-at-uni-bielefeld.de
Date: Tue, 21 Dec 2010 03:00:23 -0600
Subject: [Microscopy] Re: changing oil in the Pfeiffer-Balzer 170 THP turbo pump

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi Bill,

along with the TEM came a detailed instruction brochure from PFEIFFER ,
if you can't find it contact PFEIFFER for a pdf or me offline ( changing
oil is absolute simple and easy, but you need the small white funnel
supplied with the pump)

greetings
peter heimann

} Does anyone know the protocol for changing oil in the Pfeiffer-Balzer
} 170 THP series turbo pump used on the Zeiss 902 TEM? I have the TL
} 011 oil and I know that each side gets 20 mls, but do you pour it in,
} force it in or is there a trick. It just seems to not to what to
} come out or go back in easily.
}
} Bill McManus
} Research Scientist
} Western Dairy Center
} Utah State University
} bill.mcmanus-at-usu.edu
}
}
} ==============================Original Headers==============================
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--
====================================================

Dr. Peter Heimann Raum: W7-107 / Tel.: +49-(0)521-106-5628
Universitaet Bielefeld Fax: +49-(0)521-106-5654
"Zellbiologie / Cell Biology" W7-107
33501 Bielefeld, Germany www.uni-bielefeld.de/biologie/cellbio



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From: junhe1970-at-gmail.com
Date: Tue, 21 Dec 2010 07:02:21 -0600
Subject: [Microscopy] viaWWW: SEM stardard grid

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Name: jun

Title-Subject: [Filtered] SEM stardard grid

Message: We frequently uses SEM ( Hitachi 4700, FE) to measure some
wafer cross-section features.
The measurement result is sometimes off the mark .
I wonder if some one here could recommend some fine grid (0.1 um)standard
Jun



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From: protrain-at-emcourses.com
Date: Tue, 21 Dec 2010 07:49:26 -0600
Subject: [Microscopy] viaWWW: SEM stardard grid

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi

I use a TEM carbon grating replica of a 2160 lines per mm cross grating.
This gives a 0.4629micron spacing and, if you fix it down carbon side up,
the sample makes a nice example of the effect of kV on imaged information.

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



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Title-Subject: [Filtered] SEM stardard grid

Message: We frequently uses SEM ( Hitachi 4700, FE) to measure some
wafer cross-section features.
The measurement result is sometimes off the mark .
I wonder if some one here could recommend some fine grid (0.1 um)standard
Jun



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From: pitrone-at-mpi-cbg.de
Date: Tue, 21 Dec 2010 15:57:45 -0600
Subject: [Microscopy] viaWWW: Job Opening in the Light Microscopy Facility of MPI-CBG

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Email: pitrone-at-mpi-cbg.de
Name: Peter Pitrone

Organization: Max Planck Institute for Molecular Cell Biology and Genetics

Title-Subject: [Filtered] Job Opening in the
Light Microscopy Facility of MPI-CBG

Message:
Dear friends of microscopy and imaging,

We are very actively searching for 2
microscopists / imaging specialists to join our
light microscopy facility team with immediate
effect.

see:
http://www.mpi-cbg.de/jobs/jobs-at-the-mpi-cbg/microscopistsimaging-specialists.html

In case of any questions, please feel free to contact us: peychl-at-mpi-cbg.de

Please pass on the advert to anyone who you think might be interested.

cheers

Peter Pitrone



Full Text of advert follows:

The Max Planck Institute of Molecular Cell
Biology and Genetics (MPI-CBG) in Dresden is
seeking motivated
Microscopists/Imaging Specialists, to work in its
Light Microscopy Facility (LMF).

The LMF is a large core facility enabling
different aspects of MPI-CBG researcherís imaging
projects including: design of imaging
experiments, choice of and training on suitable
imaging systems, and image processing,
visualisation and analysis. The LMF is part of a
network of core imaging facilities on the Dresden
Biopolis campus (see https://ifn.mpi-cbg.de.)

Light microscopy is one of the key methods of the
MPI-CBG. The LMF provides more than 20 advanced
imaging systems including laser scanning confocal
microscopy, two-photon microscopy, wide-field
microscopy, TIRF, SPIM and PALM/STORM. A super
resolution structured illumination (SIM) system
will be installed in 2011. The LMF team supports
approximately 270 users from over 35 countries.
More than 40 000 hours of instrument time is
booked annually. The working language of the
MPI-CBG is English.

Requirements:
We are seeking 1-2 candidates who are proactive
and service-oriented team players with excellent
interpersonal and organizational skills and who
enjoy helping people. They should have a first
degree (B.Sc.) and preferably also a doctorate
degree (Ph.D.) in biology or physics/optics, be
fluent in English and be ready to help scientists
with both trivial and advanced aspects of light
microscopy imaging projects. Experience in basic
as well as advanced light microscopy is required
and some experience with image analysis would be
a strong plus. The successful candidate(s) should
be ready to serve in a dynamic international
multi-user environment whereby four basic areas
of service must be covered: a) scientific support
of imaging projects, b) implementation of new
imaging technologies, c) maintenance and service
of current imaging systems and d) training and
teaching light microscopy. Ideally, the
candidate(s) should have experience working in an
imaging facility.

The positions are available immediately. The
initial contract is for 2 years. Compensation
will depend on the qualifications and experience
of the candidate. The Max Planck Society is
committed to employ more disabled persons. The
application of disabled persons is strongly
encouraged. The Max Planck Society is committed
to increase the number of female scientists.
Women are particularly encouraged to apply.

For informal inquiries, please contact the leader
of the LMF, Dr. Jan Peychl, email:
peychl-at-mpi-cbg.de

If you wish to apply for this position, please
send your CV, letter of application/motivation
and the contact information of three referees to:

Max Planck Institute
of Molecular Cell Biology and Genetics
Code: 2010-LMF-4100
Pfotenhauerstr. 108
01307 Dresden
Germany
or by email: 2010-LMF-4100-at-mpi-cbg.de

Deadline for the applications is January 7, 2011


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From: bard.smedsrod-at-uit.no
Date: Tue, 21 Dec 2010 15:58:35 -0600
Subject: [Microscopy] viaWWW: Need convincing arguments motivating purchase of SEM

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Email: bard.smedsrod-at-uit.no
Name: Bard Smedsrod

Organization: University of Tromso, Norway

Title-Subject: [Filtered] Need convincing arguments motivating purchase of SEM

Message: Our Medical faculty has a well equipped EM platform, with
highly skilled staff. But our SEM instrument is really old. This
means it can only produce low magnification, low resolution images of
samples. We have struggled with this "pre-historic" SEM instrument
for years, and are trying to convince our Faculty that our
researchers deserve a new high quality, high resolution SEM
instrument equipped with the latest technology. Our problem now is
that our researchers have quit using SEM because the old instrument
has either been out of work or produced images that are not good
enough to publish. After many years with access to only a mediocre
SEM instrument our researchers have forgotten about the potential
benefits of using SEM, and the motivation among our research staff to
use and therefore demand this methodology is litterally gone.
Research at our Faculty that might benefit from including SEM
technology include: Physiology (cardiophysiology), Vascular biology
(special emphasis on liver sinusoids), Microbiology (microfilm
studies, and gene transfer among bacteria), Virology, Tumor biology
(both molecular and cell/organ based), Pharmacology, Pharmacy, Cell
biology (both molecular and "classical" cell biology), Immunology (in
particular T-cell mediated responses). I am convinced that some of
these research groups would benefit greatly from including a modern,
high-end SEM instrument in their studies. My task as head of the EM
platform is to awaken and motivate potential users (who have
forgotten the potential benefits of using SEM after all those years
with access only to a museum type of SEM) to understand that they may
benefit from using SEM. This is why I contact the Microscopy
ListServer. My hope is that some of you can give me some striking and
convincing examples showing how a high quality SEM may add valuable
dimensions to scientific studies in the biomedical fields mentioned
above. I should be really thankful to get a feedback including
concrete examples (references to high quality papers)!
leaving
no doubt that a high-end SEM enables high quality research. With
such arguments I think I might motivate our researchers to exert
sufficient pressure on our Faculty to purchase a high end SEM.
Regards,
Bard Smedsrod
Professor and Head of Vascular Biology Research Group
Scientific head of Laboratory of Electron Microscopy
Department of Medical Biology
University of Tromso
NO-9037 Tromso
NORWAY
tel.: +47-77644687 / +47-99599463




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From: protrain-at-emcourses.com
Date: Wed, 22 Dec 2010 05:22:02 -0600
Subject: [Microscopy] viaWWW: Need convincing arguments motivating purchase

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Good day Bard

I do not know if you will remember me but I was in Tromso to commission two
TEM on behalf of Hitachi in the late 1960s.

I believe you are absolutely correct in leading your team towards a modern
high resolution SEM. The current batch of FEG SEM are able to carry out an
amazing range of techniques up to extremely high magnifications. Not to be
overlooked is just how efficient a modern FEG SEM is when used in the STEM
mode; amazing contrast enhancement even with unstained material. Then there
is x-ray analysis in the instrument, a much more convenient and simplistic
technique in both SEM and STEM, occurring at more advantageous take off
angles and with far less instrument interference for transmission specimens
than the TEM. Cryo techniques, variable pressure, new high performance
backscattered electron detectors and of course the superb performance at
very low accelerating voltages, make these instruments unbelievably flexible
in their application

I could go on and on about the revolution that FEG SEM have stimulated.
Much of my consultancy is involved with these instruments at this time, in
all corners of the world they are the cause of a refocus on scanning
electron microscopy as many move away from TEM purchases!

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com



-----Original Message-----
X-from: bard.smedsrod-at-uit.no [mailto:bard.smedsrod-at-uit.no]
Sent: 21 December 2010 21:59
To: protrain-at-emcourses.com

This Question/Comment was submitted to the Microscopy Listserver
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Email: bard.smedsrod-at-uit.no
Name: Bard Smedsrod

Organization: University of Tromso, Norway

Title-Subject: [Filtered] Need convincing arguments motivating purchase of
SEM

Message: Our Medical faculty has a well equipped EM platform, with
highly skilled staff. But our SEM instrument is really old. This
means it can only produce low magnification, low resolution images of
samples. We have struggled with this "pre-historic" SEM instrument
for years, and are trying to convince our Faculty that our
researchers deserve a new high quality, high resolution SEM
instrument equipped with the latest technology. Our problem now is
that our researchers have quit using SEM because the old instrument
has either been out of work or produced images that are not good
enough to publish. After many years with access to only a mediocre
SEM instrument our researchers have forgotten about the potential
benefits of using SEM, and the motivation among our research staff to
use and therefore demand this methodology is litterally gone.
Research at our Faculty that might benefit from including SEM
technology include: Physiology (cardiophysiology), Vascular biology
(special emphasis on liver sinusoids), Microbiology (microfilm
studies, and gene transfer among bacteria), Virology, Tumor biology
(both molecular and cell/organ based), Pharmacology, Pharmacy, Cell
biology (both molecular and "classical" cell biology), Immunology (in
particular T-cell mediated responses). I am convinced that some of
these research groups would benefit greatly from including a modern,
high-end SEM instrument in their studies. My task as head of the EM
platform is to awaken and motivate potential users (who have
forgotten the potential benefits of using SEM after all those years
with access only to a museum type of SEM) to understand that they may
benefit from using SEM. This is why I contact the Microscopy
ListServer. My hope is that some of you can give me some striking and
convincing examples showing how a high quality SEM may add valuable
dimensions to scientific studies in the biomedical fields mentioned
above. I should be really thankful to get a feedback including
concrete examples (references to high quality papers)!
leaving
no doubt that a high-end SEM enables high quality research. With
such arguments I think I might motivate our researchers to exert
sufficient pressure on our Faculty to purchase a high end SEM.
Regards,
Bard Smedsrod
Professor and Head of Vascular Biology Research Group
Scientific head of Laboratory of Electron Microscopy
Department of Medical Biology
University of Tromso
NO-9037 Tromso
NORWAY
tel.: +47-77644687 / +47-99599463




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==============================Original Headers==============================
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From: oshel1pe-at-cmich.edu
Date: Wed, 22 Dec 2010 08:38:11 -0600
Subject: [Microscopy] viaWWW: Need convincing arguments motivating purchase of SEM

Contents Retrieved from Microscopy Listserver Archives
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Bard,

One excellent reason is that a modern FE-SEM can reveal the true state of cellular extensions - microvilli, pseudopods of various kinds, nanotubes connecting cells and so on. These are very difficult to study in a TEM, and many of their important features are sub-resolution in the light microscope. These are important functional features of cells.

An even better reason is immunolabelling of cell surface features. Receptor distributions, numbers, associations between and within receptor types and so on. These are very difficult or worse to study in a TEM, since one is looking at a 60-100 nm section through the plasmalemma, so receptors are easily missed. In a SEM, the entire cell surface can be imaged, and all of the receptors labelled and counted, or whatever.
For a quick example, check the U. Wisconsin BBPIC web page on immunolabelling:
http://www.ansci.wisc.edu/facstaff/Faculty/pages/albrecht/albrecht_web/Programs/microscopy/colloid.html
There is an image gallery at the bottom of the page.
Further, by using colloidal metals of different shapes, one can do multiple-labelling for SEM, and if their atomic number is sufficiently different, potentially the labels can be distinguished by a good backscatter detector.

Phil

Philip Oshel
Microscopy Facility Supervisor
Department of Biology
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576




-----Original Message-----
X-from: bard.smedsrod-at-uit.no [mailto:bard.smedsrod-at-uit.no]
Sent: Tue 10/12/21 17:02
To: Oshel, Philip Eugene

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at
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Email: bard.smedsrod-at-uit.no
Name: Bard Smedsrod

Organization: University of Tromso, Norway

Title-Subject: [Filtered] Need convincing arguments motivating purchase of SEM

Message: Our Medical faculty has a well equipped EM platform, with
highly skilled staff. But our SEM instrument is really old. This
means it can only produce low magnification, low resolution images of
samples. We have struggled with this "pre-historic" SEM instrument
for years, and are trying to convince our Faculty that our
researchers deserve a new high quality, high resolution SEM
instrument equipped with the latest technology. Our problem now is
that our researchers have quit using SEM because the old instrument
has either been out of work or produced images that are not good
enough to publish. After many years with access to only a mediocre
SEM instrument our researchers have forgotten about the potential
benefits of using SEM, and the motivation among our research staff to
use and therefore demand this methodology is litterally gone.
Research at our Faculty that might benefit from including SEM
technology include: Physiology (cardiophysiology), Vascular biology
(special emphasis on liver sinusoids), Microbiology (microfilm
studies, and gene transfer among bacteria), Virology, Tumor biology
(both molecular and cell/organ based), Pharmacology, Pharmacy, Cell
biology (both molecular and "classical" cell biology), Immunology (in
particular T-cell mediated responses). I am convinced that some of
these research groups would benefit greatly from including a modern,
high-end SEM instrument in their studies. My task as head of the EM
platform is to awaken and motivate potential users (who have
forgotten the potential benefits of using SEM after all those years
with access only to a museum type of SEM) to understand that they may
benefit from using SEM. This is why I contact the Microscopy
ListServer. My hope is that some of you can give me some striking and
convincing examples showing how a high quality SEM may add valuable
dimensions to scientific studies in the biomedical fields mentioned
above. I should be really thankful to get a feedback including
concrete examples (references to high quality papers)!
leaving
no doubt that a high-end SEM enables high quality research. With
such arguments I think I might motivate our researchers to exert
sufficient pressure on our Faculty to purchase a high end SEM.
Regards,
Bard Smedsrod
Professor and Head of Vascular Biology Research Group
Scientific head of Laboratory of Electron Microscopy
Department of Medical Biology
University of Tromso
NO-9037 Tromso
NORWAY
tel.: +47-77644687 / +47-99599463




Login Host: 129.242.155.15
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23, 34 -- From oshel1pe-at-cmich.edu Wed Dec 22 08:38:11 2010
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From: William.F.Tivol-at-aero.org
Date: 12/22/2010 06:49 AM
Subject: [Microscopy] RE: viaWWW: Need convincing arguments

Contents Retrieved from Microscopy Listserver Archives
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Dear Bard & Phil,
I have to put in a plug for cryoTEM to look at cellular
extensions, and, for that matter, anything throughout the cell. With
tomography, one can get 3D info about cilia, pseudopods, nanotubes, etc.
One thing you can point out, Brad, is that the equipment for cryoTEM is
much more expensive than SEM.
Yours,
Bill



X-from: oshel1pe-at-cmich.edu
To: William.F.Tivol-at-aero.org


One excellent reason is that a modern FE-SEM can reveal the true state of
cellular extensions - microvilli, pseudopods of various kinds, nanotubes
connecting cells and so on. These are very difficult to study in a TEM,
and many of their important features are sub-resolution in the light
microscope. These are important functional features of cells.

An even better reason is immunolabelling of cell surface features.
Receptor distributions, numbers, associations between and within receptor
types and so on. These are very difficult or worse to study in a TEM,
since one is looking at a 60-100 nm section through the plasmalemma, so
receptors are easily missed. In a SEM, the entire cell surface can be
imaged, and all of the receptors labelled and counted, or whatever.
For a quick example, check the U. Wisconsin BBPIC web page on
immunolabelling:
http://www.ansci.wisc.edu/facstaff/Faculty/pages/albrecht/albrecht_web/Programs/microscopy/colloid.html

There is an image gallery at the bottom of the page.
Further, by using colloidal metals of different shapes, one can do
multiple-labelling for SEM, and if their atomic number is sufficiently
different, potentially the labels can be distinguished by a good
backscatter detector.


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From: Frank_Karl-at-lincolnelectric.com
Date: Wed, 22 Dec 2010 11:07:02 -0600
Subject: [Microscopy] 1937 article on microscopy

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Just for giggles. I found this article on my lunch hour.

http://www.popsci.com/archive-viewer?id=dyYDAAAAMBAJ&pg=48&query=turn+the
+microscope+on+your+christmas+tree

It's from a Jan 1937 Popular Science article on microscopes and Christmas
Trees. It reminds me of why microscopy is both a hobby and profession for
me. I hope you enjoy it.

Merry Christmas everyone!

Frank

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From: germpore-at-sonic.net
Date: Wed, 22 Dec 2010 13:01:26 -0600
Subject: [Microscopy] Sensor care in microscope cameras + Low pass/anti-aliasing filter?

Contents Retrieved from Microscopy Listserver Archives
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I've been reading up on digital camera maintenance, and most of the
sites/literature on the topic concerns digital SLRs. I've been looking
at this site in particular:

http://www.cleaningdigitalcameras.com/

I was wondering, first, if microscope cameras, such as the Zeiss
Axiocam, Photometrics CoolSnap, or QImaging QICAM, have an anti-
aliasing (aka "low-pass") filter in front of the actual sensor the way
that digital SLRs do. If so, do microscope camera occasionally require
the same "sensor cleaning" that digital SLRs do?

The only time I've looked straight inside of a microscope camera was
when changing the mount on a Zeiss Axiocam, and in that case, it seems
like the sensor was very exposed, with no glass between the front of
the camera and the sensor. If this typical of microscope cameras, what
should one do if dust gets on the sensor? Obviously, cleaning the
actual sensor is a great deal more risky proposition (if it is doable
at all) than typical "sensor cleaning" that actually means cleaning
the anti-aliasing filter.

Obviously, microscope cameras generally stay mounted in a lens-down
position and without regular changing of lenses, and so have the
inside of the camera much less exposed to dust and dirt exposure than
would be typical of an SLR, hence the need for cleaning should be much
less frequent, in any event.

Also, if there's a website or publication that gives general methods
for care and maintenance of microscope cameras, please let me know.

Peter

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From: NEERAJG-at-clemson.edu
Date: Wed, 22 Dec 2010 13:09:32 -0600
Subject: [Microscopy] 1937 article on microscopy

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html


} Cool article! I think such exercises are crucial for kids, that's how I got hooked on microscopy, got my first toy microscope when I was in 2nd grade and looking at things like leaves, ants and onion peels was beyond fascinating!
}
} Happy Holidays Everyone!
}
} Neeraj.
}
}
}
} On Dec 22, 2010, at 12:11 PM, "Frank_Karl-at-lincolnelectric.com" {Frank_Karl-at-lincolnelectric.com} wrote:
}
} }
} }
} }
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} }
} } Just for giggles. I found this article on my lunch hour.
} }
} } http://www.popsci.com/archive-viewer?id=dyYDAAAAMBAJ&pg=48&query=turn+the
} } +microscope+on+your+christmas+tree
} }
} } It's from a Jan 1937 Popular Science article on microscopes and Christmas
} } Trees. It reminds me of why microscopy is both a hobby and profession for
} } me. I hope you enjoy it.
} }
} } Merry Christmas everyone!
} }
} } Frank
} }
} } --
} } *************************************************************
} } Note:
} } The information contained in this message may be
} } privileged and confidential and protected from disclosure. If
} } the reader of this message is not the intended recipient, or
} } an employee or agent responsible for delivering this message
} } to the intended recipient, you are hereby notified that any
} } dissemination, distribution or copying of this communication
} } is strictly prohibited. If you have received this
} } communication in error, please notify us immediately by
} } replying to the message and deleting it from your computer.
} } Thank you,
} } The Lincoln Electric Company
} } **************************************************************
} }
} }
} } ==============================Original Headers==============================
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From: cosgrove-at-spotimaging.com
Date: Wed, 22 Dec 2010 16:30:03 -0600
Subject: [Microscopy] Re: Sensor care in microscope cameras

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http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Peter,
One of the scopes I worked on had a CCD mounted just below the
film camera, which the user routinely collected data with. When the
resolution of the CCD started to deteriorate, it was discovered that there
was contamination on the sensor, so the service person gently washed the
sensor with ethanol. This restored most of the resolution to the CCD
images. The sensor surface is very delicate, and, unless you use film a
lot, there is not likely to be too much contamination, but if you notice
that the resolution of the CCD starts to deteriorate, the sensor may need
cleaning. I would not recommend this procedure as a part of routine
maintenance, and I would definitely let the professionals handle it.
Yours,
Bill



X-from: germpore-at-sonic.net


Hi Peter,

I don't have an online resource to point you to, but here are a few
hopefully useful points that we typically give our customers:

1) Scientific microscopy cameras do not introduce an anti-aliasing filter
into the system and therefore do not need cleaning of this element.
 
2) Scientific cameras usually have a front chamber window to keep dust out
of the sealed chamber that houses the sensor.  Simple sealed chambers are
sealed against dust but not moisture and are used for non-cooled cameras.
Stronger seals prevent both dust and moisture infiltration and are used on
cooled cameras to prevent moisture condensation on the sensor.
 
3) Some cameras sensors are provided in a "windowless" package exposing the
surface of the sensor to the environment present in the sealing chamber
itself. This configuration of camera should only be considered for the most
demanding of applications due to the risk of sensor contamination. Cleaning
of the sensor surface is only possible on non-micro-lens designs, and then
only should only be attempted as a last resort as the surface and bonding
wire connections are extremely delicate.
 
4) Cleaning should be limited to the outside of the sealing chamber window. 
Breaking the seal to the sealing chamber may result in dust particles being
introduced into the chamber if not performed in clean hood environment.
Additionally, breaking the seal on cooled cameras may result in moisture
infiltration and saturation of the molecular sieve resulting in condensation
on the sensor.

Best regards,

Sarah Cosgrove
SPOT Imaging Solutions, a division of Diagnostic Instruments, Inc.
www.spotimaging.com




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From: lbrako-at-yahoo.com
Date: Wed, 22 Dec 2010 17:55:17 -0600
Subject: [Microscopy] viaWWW: TEM need help with staining

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Email: lbrako-at-yahoo.com
Name: lawrence Brako

Organization: morehouse school of medicine

Title-Subject: [Filtered] TEM need help with staining

Message: Hi All,
Has anyone else ever had problems staining the
melanosomes of RPE cells (retinal pigment
epithelial) for TEM? They usually appear very
dark/black, and I had them that way several years
ago, but now in all my images they appear
consistently gray regardless of how long I
poststain with 8% Uranyl Acetate and Reynoldsí
lead citrate. Every other structure seems to
stain pretty much okay.
I use routine TEM protocol, fresh 2.5% Glut.,
wash, fresh 2%OsO4 for 1hr, wash, en bloc with
0.5% UAc, dehydration, resin mix(Epon 812)
infiltration, etc etc. I just cannot figure out
where the problem originates. Any takers?? Thank
you kindly,


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From: dlowry-at-asu.edu
Date: Wed, 22 Dec 2010 17:56:09 -0600
Subject: [Microscopy] viaWWW: irregularity in TEM alignment procedure

Contents Retrieved from Microscopy Listserver Archives
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Email: dlowry-at-asu.edu
Name: David Lowry

Organization: Arizona State University

Title-Subject: [Filtered] irregularity in TEM alignment procedure

Message: recently when doing basic alignments for our TEM a problem
was noticed. After setting z-height of specimen and adjusting pivot
points and lens-current centering in normal manner, the
voltage-center adjustment exhibits extreme deviation. After
re-focusing the image (} 100k x mag) and engaging voltage-center
function, the focus of image becomes extremely far off, so much so
that detail cannot be seen to make the adjustment. The obj lens has
to be defocused approx 50um in order to bring the image back to
focus, and the beam moves off-center on the viewing screen. Once in
focus the adjustment can be made, but going back and repeating the
entire align procedure produces the same deviation.

we are having difficulty envisioning what is occurring here. Some
electronic boards were recently replaced on this TEM for an unrelated
issue, not sure if this may be the cause of the current problem.
Nothing in the column or stage has been disassembled.

thanks for any help or suggestions

Login Host: 149.169.143.79
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From: W.Muss-at-salk.at
Date: Thu, 23 Dec 2010 03:43:16 -0600
Subject: [Microscopy] Re: TEM need help with staining

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear Lawrence Brako,
good morning !

Concerning your request, I guess the en-bloc-staining with 0.5% UO2Ac, followed then by 8% UO2Ac (I suppose that solution to be hydrous?) and Reynolds lead citrate on ultrathin sections might be the problem.

There have been similar questions to the listserver within the last years.... IF I am able to summarize for you that postings (would you accept also forwarding the specific mail-messages as .msg to your mail-box??) within the next days or at least until first week of the new year could this perhaps help you somehow?.

Just being adherent to the technical problem:
a) have you stained your melanosome preps former ago also using en bloc UO2Ac-stain or only "plain ultrathin section double staining as usual? Which result?
b) which preparatory steps do you use before UO2-Ac en bloc (i.e. out of OsO4 - A.dest-A.dest- hydrous (?) 0.5% UO2 Ac.... or e.g. out of OSO4, washing in maleate buffer (with correction of pH) and UO2Ac in Maleate buffer/correct pH adjusted....washes in Maleate buffer or A.dest...)

c) Section staining with 8% UO2 Ac (hydrous?) and for how long? (UO2Ac as well as Pbcitrate) Why 8% UO2 Ac ?

d) have you thought about "triple staining" your ultrathins, e.g.
Preincubation of sections in 0.05% hydrous Tannic acid (low molecular weight)
[dissolve by ultrasonication, filter then, suck into a [previously cleaned] discardable single-use syringe,
fitted with a Millipore or other filter 0.45µm. Use the solution dropwise on e.g. parafilm surface] ==}
incubation (grids inverted) 5-8 min at room temperature ("darkness" doesn't "harm"), wash (2-3x a.dest. jet),
suck off water from grid with filter paper as usual, let dry, then proceed with UO2ac(protection from light)
and Pb-citrate as usual.


My personal experience some years ago was that UO2Ac-section staining AFTER en-bloc-Staining with UO2Ac always did result in an inferior staining intensity in most structures known to bind UO2 Ac (as this was also the case for UO2Ac (1% methanolic[100% MetOH] + "trace" of acetic acid)- section staining and too long Pb-citrate staining....
Interestingly, when using the so called "NSG-UO2Ac-staining = Uranaffine reaction method" (for specific localization of neurosecretory granules in brain or nerve tissues), which in fact consist also in an en-bloc staining of tissue slices, you'll get specific contrast for those NSG. "double staining" the sections with UO2Ac in the first place decreases also the contrast of those normally specifically stained NSG's....(:-))
Hope these explanations for now are of value for you and solving your problem,

Merry Christmas,
Happy Holidays, and best wishes for a Happy, healthy, joyous, successful and prosperous 2011,

regards


Wolfgang MUSS
Head EM-Lab
Univ.Inst.Pathology
SALK-LKH (Gen.Hospital)
SALZBURG, AUSTRIA

} -----Ursprüngliche Nachricht-----
} Von: lbrako-at-yahoo.com [mailto:lbrako-at-yahoo.com]
} Gesendet: Donnerstag, 23. Dezember 2010 00:59
} An: Muß Wolfgang
} Betreff: [Microscopy] TEM need help with staining
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} Email: lbrako-at-yahoo.com
} Name: lawrence Brako
} Organization: morehouse school of medicine
} Title-Subject: TEM need help with
} staining
} Message:
}
} Hi All,
} Has anyone else ever had problems staining the melanosomes of RPE cells
} (retinal pigment epithelial) for TEM?
}
} They usually appear very dark/black, and I had them that way several
} years ago, but now in all my images they appear consistently gray
} regardless of how long I poststain with 8% Uranyl Acetate and Reynolds'
} lead citrate. Every other structure seems to stain pretty much okay.
}
} I use routine TEM protocol, fresh 2.5% Glut., wash, fresh 2%OsO4 for
} 1hr, wash, en bloc with 0.5% UAc, dehydration, resin mix(Epon 812)
} infiltration, etc etc.
}
} I just cannot figure out where the problem originates.
} Any takers?? Thank you kindly,




} Morehouse School of Medicine
} 720 Westview Drive SW Atlanta, GA 30310-1495
} www.msm.edu/
}
}
} Login Host: 192.83.232.85
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From: protrain-at-emcourses.com
Date: Thu, 23 Dec 2010 10:14:25 -0600
Subject: [Microscopy] viaWWW: irregularity in TEM alignment procedure

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi

This sounds very strange!

The voltage and current centres should be within 4 microns of each other,
their alignments achieved by tilting the total illuminating system in
relation to the objective lens axis. I see no clear reason for the problem
that you have.

My first action would be to check to see if your eucentric point setting
requires an objective lens current that is identical to what you have had in
the past. A possible problem being that you may not be using your normal
focal currents? Was the current alignment far out when you first checked it
prior to the problem being discovered? Try a lower accelerating voltage
just in case the problem is high voltage related?

My basic thought would be to look for something unusual with your settings;
double check gun alignments and illumination system alignments.

Good luck but come back with any further information.

Steve

Steve Chapman FRMS
Senior Consultant Protrain
For consultancy and training in electron microscopy world wide
Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Cell +44 (0)7711 606967
www.emcourses.com





-----Original Message-----
X-from: dlowry-at-asu.edu [mailto:dlowry-at-asu.edu]
Sent: 22 December 2010 23:57
To: protrain-at-emcourses.com

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: dlowry-at-asu.edu
Name: David Lowry

Organization: Arizona State University

Title-Subject: [Filtered] irregularity in TEM alignment procedure

Message: recently when doing basic alignments for our TEM a problem
was noticed. After setting z-height of specimen and adjusting pivot
points and lens-current centering in normal manner, the
voltage-center adjustment exhibits extreme deviation. After
re-focusing the image (} 100k x mag) and engaging voltage-center
function, the focus of image becomes extremely far off, so much so
that detail cannot be seen to make the adjustment. The obj lens has
to be defocused approx 50um in order to bring the image back to
focus, and the beam moves off-center on the viewing screen. Once in
focus the adjustment can be made, but going back and repeating the
entire align procedure produces the same deviation.

we are having difficulty envisioning what is occurring here. Some
electronic boards were recently replaced on this TEM for an unrelated
issue, not sure if this may be the cause of the current problem.
Nothing in the column or stage has been disassembled.

thanks for any help or suggestions

Login Host: 149.169.143.79
---------------------------------------------------------------------------

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From: sawyert-at-science.oregonstate.edu
Date: Thu, 23 Dec 2010 18:04:57 -0600
Subject: [Microscopy] viaWWW: Iodine stain

Contents Retrieved from Microscopy Listserver Archives
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Email: sawyert-at-science.oregonstate.edu
Name: Teresa Sawyer

Organization: Oregon State University

Title-Subject: [Filtered] Iodine stain

Message: Our Plant Anatomy class needs a stain for starch detection.
IKI (potassium iodide + iodine) has been used in the past but they
are having difficulty finding this stain. Any help location this
stain or an alternate stain would be great.

Teresa

Login Host: 128.193.226.197
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From: rosemary.white-at-csiro.au
Date: Thu, 23 Dec 2010 18:58:39 -0600
Subject: [Microscopy] Re: viaWWW: Iodine stain

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hi Teresa,
Standard iodine stain:

Dissolve 2 g potassium iodide salts in 100 ml water, then add 0.2 g iodine
crystals.

That's it, you'll get a dark brown solution. This is quite strong, and for
some tissue can be diluted 1 in 10 or more.

Note that iodine is poisonous, don't inhale the iodine vapours from the
crystals.

By the way, for plant anatomy the book "Teaching Plant Anatomy" by the L &
CA Peterson and LH Melville is terrific, all fresh tissue observations
beautifully illustrated and with basic staining protocols at the end.

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au


On 24/12/10 11:10 AM, "sawyert-at-science.oregonstate.edu"
{sawyert-at-science.oregonstate.edu} wrote:

}
}
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} Email: sawyert-at-science.oregonstate.edu
} Name: Teresa Sawyer
}
} Organization: Oregon State University
}
} Title-Subject: [Filtered] Iodine stain
}
} Message: Our Plant Anatomy class needs a stain for starch detection.
} IKI (potassium iodide + iodine) has been used in the past but they
} are having difficulty finding this stain. Any help location this
} stain or an alternate stain would be great.
}
} Teresa
}
} Login Host: 128.193.226.197
} ---------------------------------------------------------------------------
}
} ==============================Original Headers==============================
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==============================Original Headers==============================
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From: germpore-at-sonic.net
Date: Thu, 23 Dec 2010 19:20:45 -0600
Subject: [Microscopy] Re: Iodine stain

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

sawyert-at-science.oregonstate.edu wrote:

} Name: Teresa Sawyer
}
} Organization: Oregon State University
}
} Title-Subject: [Filtered] Iodine stain
}
} Message: Our Plant Anatomy class needs a stain for starch detection.
} IKI (potassium iodide + iodine) has been used in the past but they
} are having difficulty finding this stain. Any help location this
} stain or an alternate stain would be great.

Hi –

I'm unclear as to why you would be having difficulty obtaining IKI.
The standard IKI solution is Lugol's iodine, and not only is that
available through sources like Carolina and VWR, I've even it sold
over-the-counter through "green" pharmacies, and over Amazon.com.
Lugol's is good enough for simple starch detection and there's no
reason you should have difficulty obtaining either it, or stock iodine
and potassium iodide to make up your own. It would surprise me, in
fact, if your department's stockroom didn't have these already.

The difficult-to-find iodine solution you might be thinking of is
Melzer's, in which the IKI is in a chloral hydrate/aqueous solution
rather than a straight aqueous solution. The chloral hydrate gives the
solution additional clearing properties which are useful with heavily
pigmented tissue. However, due to the Schedule IV status of chloral
hydrate and the DEA's stepped up enforcement of licensing of such
compounds, it's difficult to get the ingredients to make unless one is
willing to go through DEA paperwork. I posted a question about
possible Melzer's substitutes to this list only a month ago, because
I've been up against the same problem.

However, being a bit familiar with your department (Botany & Plant
Pathology -at-OSU, right?), I know there are a number of mycologists in
that department (such as Joey Spatafora) who use Melzer's as a
standard tool. If Melzer's is in fact what you need, I would talk to
those people about where to obtain it.

Peter



==============================Original Headers==============================
9, 22 -- From germpore-at-sonic.net Thu Dec 23 19:20:45 2010
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From: rosemary.white-at-csiro.au
Date: Thu, 23 Dec 2010 19:51:33 -0600
Subject: [Microscopy] Iodine stain

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hmm, for a prac class I'd be reluctant to have students using a chloral
hydrate based stain.

If the tissue is highly pigmented, I wonder if bleaching in NaOH or KOH
first - from 0.1 to 1 M, depending on degree of pigmentation and toughness
of tissue, or even with dilute household bleach, might help?

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au


On 24/12/10 12:25 PM, "germpore-at-sonic.net" {germpore-at-sonic.net} wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} sawyert-at-science.oregonstate.edu wrote:
}
} } Name: Teresa Sawyer
} }
} } Organization: Oregon State University
} }
} } Title-Subject: [Filtered] Iodine stain
} }
} } Message: Our Plant Anatomy class needs a stain for starch detection.
} } IKI (potassium iodide + iodine) has been used in the past but they
} } are having difficulty finding this stain. Any help location this
} } stain or an alternate stain would be great.
}
} Hi ­
}
} I'm unclear as to why you would be having difficulty obtaining IKI.
} The standard IKI solution is Lugol's iodine, and not only is that
} available through sources like Carolina and VWR, I've even it sold
} over-the-counter through "green" pharmacies, and over Amazon.com.
} Lugol's is good enough for simple starch detection and there's no
} reason you should have difficulty obtaining either it, or stock iodine
} and potassium iodide to make up your own. It would surprise me, in
} fact, if your department's stockroom didn't have these already.
}
} The difficult-to-find iodine solution you might be thinking of is
} Melzer's, in which the IKI is in a chloral hydrate/aqueous solution
} rather than a straight aqueous solution. The chloral hydrate gives the
} solution additional clearing properties which are useful with heavily
} pigmented tissue. However, due to the Schedule IV status of chloral
} hydrate and the DEA's stepped up enforcement of licensing of such
} compounds, it's difficult to get the ingredients to make unless one is
} willing to go through DEA paperwork. I posted a question about
} possible Melzer's substitutes to this list only a month ago, because
} I've been up against the same problem.
}
} However, being a bit familiar with your department (Botany & Plant
} Pathology -at-OSU, right?), I know there are a number of mycologists in
} that department (such as Joey Spatafora) who use Melzer's as a
} standard tool. If Melzer's is in fact what you need, I would talk to
} those people about where to obtain it.
}
} Peter
}
}
}
} ==============================Original Headers==============================
} 9, 22 -- From germpore-at-sonic.net Thu Dec 23 19:20:45 2010
} 9, 22 -- Received: from qmta03.emeryville.ca.mail.comcast.net
} (qmta03.emeryville.ca.mail.comcast.net [76.96.30.32])
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} 9, 22 -- From: Peter Werner {germpore-at-sonic.net}
} 9, 22 -- To: Microscopy-at-microscopy.com, sawyert-at-science.oregonstate.edu
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} delsp=yes
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} 9, 22 -- Date: Thu, 23 Dec 2010 17:20:30 -0800
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From: germpore-at-sonic.net
Date: Thu, 23 Dec 2010 20:53:24 -0600
Subject: [Microscopy] Re: Iodine stain

Contents Retrieved from Microscopy Listserver Archives
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rosemary.white-at-csiro.au wrote:

} Hmm, for a prac class I'd be reluctant to have students using a
} chloral
} hydrate based stain.

Melzer's is regularly used in introductory mycology classes. I've
never heard of anybody having problems with handling it.

} If the tissue is highly pigmented, I wonder if bleaching in NaOH or
} KOH
} first - from 0.1 to 1 M, depending on degree of pigmentation and
} toughness
} of tissue, or even with dilute household bleach, might help?

I'm not sure how that might affect the solubility of the iodine, or
might even be reactive with that and the potassium iodide. I've heard
one recommendation for Lactophenol/IKI as a substitute for Melzer's,
at least in terms of having an IKI + clearing solution. However, it
does not consistently produce the same staining reactions as Melzer's,
which in terms of fungal taxonomy at least, makes it less useful.
Mycology has many years of recorded reactions of spores and tissues to
Melzer's specifically, and these are key points of identification for
some taxa.

Again, for the purposes of the plant anatomy class that the original
poster was describing, I doubt anything beyond simple Lugol's would be
needed, though the fact that they're having a hard time getting more
of the compound makes me suspect the class may have previously used
Melzer's rather than Lugol's.

Peter

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From: PWebster-at-hei.org
Date: Thu, 23 Dec 2010 21:23:05 -0600
Subject: [Microscopy] RE: Need convincing arguments motivating purchase of SEM

Contents Retrieved from Microscopy Listserver Archives
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This is an interesting request in that many years ago I would have replied with the retort that biologists have no use for scanning electron microscopy. I was brought up with the idea that all useful information can be obtained using thin sections and a transmission EM.

However, I run a shared imaging facility and was instructed to get a scanning EM as part of the initial set-up. We had money available for a good quality TEM and associated specimen preparation equipment (especially a cryo-ultramicrotome) so it did not bother me that we would have a scanning EM sitting unused. How wrong I was.

First, let me say that my scanning EM is now 11 years old and has more use than the TEM sitting in the next room. There are many reasons for why this is, but the reason most often quoted by my users is that it is easy to prepare specimens for the machine, and also the most easy to use. Having trained how to operate confocal microscopes I can see how the ease of use compares.

We can easily train users to operate the microscope because it is fully controlled by software and mouse clicks. Every year we host one or two summer students who are often in their final year of high school, and they take to the SEM rapidly. within a week they are able to train others and their enthusiasm turns them into effective ambassadors for what we do.

The samples we have examined in the scanning EM have been multiple and varied. We first used it to look at the organ of Corti in the mammalian inner ear, a particularly well organized structure that offers much information on mammalian hearing. As I work in a a hearing research institute it is important that we are able to look at the inner ear structures. One particular structure that is difficult to image in the TEM is the thin protein bridge between the stereocillia (sticking out of the top of the sensory hair cells). Being able to obtain the resolution that reveals the individual proteins on this bridge is amazing.

We have had users who image frog ears, zebrafish, individual neurons growing in culture, extracted neurons with myosin-decorated actin filaments, cultured cells, vibratome sections, dissected tissues, nanoparticles, synthetic organs formed on polymer scaffolds, and much more.

Another use of the SEM that I had not initially foreseen was being able to image bacteria attaching to mammalian cell surfaces. Revealing how the cells respond to the bacteria was fascinating, as was the process by which the bacteria are engulfed by the cells. Of course it didn't take us long to move on to imaging activated and non-activated macrophages. These images drew in graduate students who wanted to learn how to obtain similar images.

Our study of extracellular bacteria took us in a slightly different direction and resulted in us studying bacterial biofilms. The SEM was an essential part of us revealing complex 3-D structures formed by aggregations of extracellular bacterial proteins. The biofilm structures that we are imaging point to the possibility that bacteria may have an existence that is more than as individual cells.

Our 11-year old SEM is now looking very old and we are in the process of discussing how we could replace it. First we will have to find the money, but then we will have to decide in which direction we would like to go if we do buy a new machine.

We have been discussing the different microscope options available and have been talking with the microscope manufacturers. From these discussions it is clear that we need to upgrade as soon as possible. The options available for biologists are amazingly varied and useful.

Talk with SEM suppliers and you will find an impressive amount of technological development. Modern scanning EMs offer different combinations of high or low vacuum, high resolution and an impressive amount of novel hardware and software. How these tools are used seems to be limited only by the imagination of the operator,

For example, it is possible to perform automated serial sectioning of resin-embedded tissues and cells using a scanning EM. The development of an in-microscope ultramicrotome by Denk and Horstmann (PLoS Biol. 2004 Nov;2(11):e329. Epub 2004 Oct 19. Serial block-face scanning electron microscopy to reconstruct three-dimensional tissue nanostructure) will greatly assist anyone wanting to produce 3-D reconstructions through large volumes of tissue. Basically, the embedded ultramicrotome slices a thin slice from the resin block and the SEM automatically images the block surface using the back-scatter electron detector. This attachment is on sale commercially as the 3-View system.

A technique called “array tomography” also makes use of resin embedded sections, but in this instance, the sections are first mounted on glass slides so that they can be imaged by confocal microscopy first (search PubMed using “array tomography smith” for details). Array tomography makes it possible to label thin sections with antibodies and fluorescent dyes and then overlay SEM images of the same regions to produce reference space data (instead of the black space that the confocal presents).

Dual beam microscopes offer an alternative approach to “sectioning” biological material while imaging it at high resolution. Imagine being able to slice through a cell and image the Golgi complex, “look for” endocytosed markers inside cells in real time, or look for intracellular viral particles.

Paul Walther in Germany (Höhn et al. Histochem Cell Biol. 2010 Nov 27. Preparation of cryofixed cells for improved 3D ultrastructure with scanning transmission electron tomography) has been quietly pioneering the use of cryo-specimen stages on SEMs to image the inside of cells. One of his approaches is to cryo-plane specimens (cryo-section with a diamond knife) and image the cut surfaces in the SEM, and collecting images as frozen water gradually sublimates in the high vacuum of the microscope. Being able to prepare specimens for SEM using high pressure freezing and freeze substitution is an advantage for imaging with only minor ultrastructural changes.

Some attempts have been made to image colloidal gold particles in the SEM and I think this approach will expand once the methods have been worked out. Imagine watching the aggregation of gold particles on cell surfaces, and their internalization into coated pits. Similarly, it should be possible to image isolated organelles and examine vesicle fusion, or attachment to actin filaments or microtubules.


Modern SEMs offer a wide range of options. Variable pressure machines make it possible to image biological material without metal coating, high resolution machines offer the ability to “look at” high resolution structures on cell surfaces as well as intracellular membranes. Dual beam machines and embedded tools (such as ultramicrotomes or AFMs) give us ways of manipulating specimens that were previously not possible (or not easy). Combining light microscopy with the scanning EM (in situ hybridization or immunolabeling – e.g. Schaudinn et al. J Microsc. 2009 Aug;235(2):124-7. Imaging of endodontic biofilms by combined microscopy (FISH/cLSM - SEM) reveals structural detail not available if only one of these imaging modes is used.

In conclusion, although there are many technical reasons why a biology department should have a scanning EM available for use (in addition to at least one high-end TEM), perhaps the most compelling reason to have an SEM available is that post-docs and graduate students will always find surprising new ways of imaging cells and tissues.

Happy Holidays,

Paul Webster
House Ear Institute
2100 W 3rd St.
Los Angeles
CA 90057
(213) 273 8026





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From: rcommon-at-msu.edu
Date: Thu, 23 Dec 2010 21:43:13 -0600
Subject: [Microscopy] Iodine stain

Contents Retrieved from Microscopy Listserver Archives
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Both KOH and houshold bleach have uses in conjunction with iodine
staining, but they are not compatible with the iodine reagent and must
be thoroughly rinsed out before applying the iodine. Strong
pretreatment with KOH can cause some materials, like cellulose, to
become more reactive to iodine. Some materials common in fungi have an
I+ red reaction to IKI without KOH pretreatment and are I+ blue after
KOH. Some pigmented tissues in fungi and lichens are reactive to
iodine, and the reactions are much easier to detect after bleaching (as
little as necessary to clear).

I believe it is unfortunate that the use of Melzer's reagent became
standard in mycology. I know of no type of reactive material (and there
are many) that react with Melzer's and no other reagent. To the extent
I have found Melzer's useful, it is in distinguishing substances that
are reactive in other reagents and negative in Melzer's. Also, the
terms "Melzer's reagent" and "Lugol's iodine" are problematic because
formulations with different iodine concentration have been referred to
for both in published literature. Since Iodine concentration can affect
results (some materials are blue, or negative, at low iodine
concentration and red at high concentration) the specific formulation of
iodine reagents used should be given in all published reports.

Dilute aqueous IKI should be all that is necessary to show starch.

Ralph Common

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From: germpore-at-sonic.net
Date: Fri, 24 Dec 2010 00:42:05 -0600
Subject: [Microscopy] Re: Iodine stain

Contents Retrieved from Microscopy Listserver Archives
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rcommon-at-msu.edu wrote:

} Both KOH and houshold bleach have uses in conjunction with iodine
} staining, but they are not compatible with the iodine reagent and must
} be thoroughly rinsed out before applying the iodine. Strong
} pretreatment with KOH can cause some materials, like cellulose, to
} become more reactive to iodine. Some materials common in fungi have
} an
} I+ red reaction to IKI without KOH pretreatment and are I+ blue after
} KOH. Some pigmented tissues in fungi and lichens are reactive to
} iodine, and the reactions are much easier to detect after bleaching
} (as
} little as necessary to clear).

Yes, now that I think about it, when I've tried using Melzer's and KOH
directly together, they immediately form a cloudy precipitate that's
no good for microscopy and probably no longer gives an iodine
reaction. (I'm not enough of a chemist to be clear about what the
precipitate consists of – potassium iodide in excess of the limit of
solubility, perhaps?)

However, I have read that pretreatment with KOH, followed by rinsing
in neutral-pH water is actually a standard for using Melzer's, as
there are some cases where some cell types in certain taxa will be
amyloid- or dextrinoid-positive with pretreatment, but negative
without it. More about this here:

www.cybertruffle.org.uk/cyberliber/59575/0003/001/0165.htm

} I believe it is unfortunate that the use of Melzer's reagent became
} standard in mycology. I know of no type of reactive material (and
} there
} are many) that react with Melzer's and no other reagent. To the
} extent
} I have found Melzer's useful, it is in distinguishing substances that
} are reactive in other reagents and negative in Melzer's.

Yes, the cells or tissue will react, usually. But sometimes not, and
often not in the same way. This article shows the result of a test of
(Langeron's modified), Melzer's, and Tincture of Iodine. The results
show numerous false positives and negatives for the non-Melzer's
solutions:

www.namyco.org/images/pdf_files/Melzer__Lugo.pdf

Unfortunately, the author used some standardized versions of each
compound in which the concentrations of I2 and KI differed. Using
several different Melzer's formulations, alongside non-Melzer's
formulations with matching concentrations of I2 & KI would have given
more definitive results.

The article also notes that Melzer's can be formulated at a
compounding pharmacy by prescription, but I have yet to approach an MD
about this.

Peter

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From: eikonika-at-otenet.gr
Date: Fri, 24 Dec 2010 03:39:44 -0600
Subject: [Microscopy] viaWWW: Need convincing arguments motivating purchase

Contents Retrieved from Microscopy Listserver Archives
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Dear Brad



In biological research, SEM is an indispensable tool, primary or adjunctive,
for any morphological / ultrastructural study. As pointed out by other
colleagues, it can give you a unique sense of the 3-D structure of the cell
surface and the architecture of the tissues. In the field of human
reproduction I am familiar with, SEM is the only means to:

-detect the pinopodes, characteristic membrane projections of the
endometrial epithelium related to receptivity

-detect salpingeal or squamous metaplasias and their extend in the
endometrial epithelium

-to resolve the 3-D structure of the zona pellucida of the oocyte

-assess the extend of cell to cell communication in cell cultures or the
cell degeneration and apoptosis in the same systems

-to detect sperm abnormalities and quantify them

-and many others, for instance the changes of the endothelium in small
arteries due to hypertension



I have published more than 30 original and review articles based partially
or entirely in SEM.(can have a look at my web site www.aim.cat -click
publications). An additional advantage is that, because SEM images are
spectacular and easily appreciated, you have a good chance to make the cover
of the journal you publish. To my mind, SEM is a necessary adjunction to any
TEM study, if you want to have the full picture of the object you study.



However, I don't agree with the need to buy a very powerful instrument for
biological applications, not even a new one. As you know, in viewing
biological specimens under SEM, you will never go above X3K, maximum X5-10K
because simply there is nothing to see further on. And SEM images in the
field of biology are not different to what was taken 40 years ago. Clearly,
it is the preparation of the soft tissue that is crucial for the quality of
the image. You mention that your old instrument does not produce decent
images; this is maybe due to a technical problem which you can fix.



I don't think that a very modern instrument will really attract more
investigators. It can work the other way around, that if you have a super
scope and still the investigators see poor images due to their poor
preparations they will permanently switch away from SEM. Probably the best
idea is to get a good old instrument, maybe from between 80- 90ies, one
instrument that was widely used and there are still many spare parts around.
In this way you will avoid the complicated electronics and software
additions that modern scopes have and that makes you depending entirely from
the company which will inevitably screw you, (as it happened to me) and in a
few years you will have to scrap the scope because no more software /
hardware support. In an old good instrument you can add an external scan
generator (I am very happy with DISS5 from the German firm
"pointelectronic") and digitize the old scope with little money and be able
to get fantastic pictures of 50 or 80 mega pixels.



I hope that my comments will be helpful to you. In the topic you discuss, as
happens everywhere, it is more the man than the machine that makes the
difference.



Happy holidays to all of you in this lovely list

yorgos



Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
eikonika-at-otenet.gr
www.aim.cat
*********************************







}
} This is an interesting request in that many years ago I would have replied
} with the retort that biologists have no use for scanning electron
} microscopy. I was brought up with the idea that all useful information can
} be obtained using thin sections and a transmission EM.
}
} However, I run a shared imaging facility and was instructed to get a
} scanning EM as part of the initial set-up. We had money available for a
} good quality TEM and associated specimen preparation equipment (especially
} a cryo-ultramicrotome) so it did not bother me that we would have a
} scanning EM sitting unused. How wrong I was.
}
} First, let me say that my scanning EM is now 11 years old and has more use
} than the TEM sitting in the next room. There are many reasons for why this
} is, but the reason most often quoted by my users is that it is easy to
} prepare specimens for the machine, and also the most easy to use. Having
} trained how to operate confocal microscopes I can see how the ease of use
} compares.
}
} We can easily train users to operate the microscope because it is fully
} controlled by software and mouse clicks. Every year we host one or two
} summer students who are often in their final year of high school, and they
} take to the SEM rapidly. within a week they are able to train others and
} their enthusiasm turns them into effective ambassadors for what we do.
}
} The samples we have examined in the scanning EM have been multiple and
} varied. We first used it to look at the organ of Corti in the mammalian
} inner ear, a particularly well organized structure that offers much
} information on mammalian hearing. As I work in a a hearing research
} institute it is important that we are able to look at the inner ear
} structures. One particular structure that is difficult to image in the TEM
} is the thin protein bridge between the stereocillia (sticking out of the
} top of the sensory hair cells). Being able to obtain the resolution that
} reveals the individual proteins on this bridge is amazing.
}
} We have had users who image frog ears, zebrafish, individual neurons
} growing in culture, extracted neurons with myosin-decorated actin
} filaments, cultured cells, vibratome sections, dissected tissues,
} nanoparticles, synthetic organs formed on polymer scaffolds, and much
} more.
}
} Another use of the SEM that I had not initially foreseen was being able to
} image bacteria attaching to mammalian cell surfaces. Revealing how the
} cells respond to the bacteria was fascinating, as was the process by which
} the bacteria are engulfed by the cells. Of course it didn't take us long
} to move on to imaging activated and non-activated macrophages. These
} images drew in graduate students who wanted to learn how to obtain similar
} images.
}
} Our study of extracellular bacteria took us in a slightly different
} direction and resulted in us studying bacterial biofilms. The SEM was an
} essential part of us revealing complex 3-D structures formed by
} aggregations of extracellular bacterial proteins. The biofilm structures
} that we are imaging point to the possibility that bacteria may have an
} existence that is more than as individual cells.
}
} Our 11-year old SEM is now looking very old and we are in the process of
} discussing how we could replace it. First we will have to find the money,
} but then we will have to decide in which direction we would like to go if
} we do buy a new machine.
}
} We have been discussing the different microscope options available and
} have been talking with the microscope manufacturers. From these
} discussions it is clear that we need to upgrade as soon as possible. The
} options available for biologists are amazingly varied and useful.
}
} Talk with SEM suppliers and you will find an impressive amount of
} technological development. Modern scanning EMs offer different
} combinations of high or low vacuum, high resolution and an impressive
} amount of novel hardware and software. How these tools are used seems to
} be limited only by the imagination of the operator,
}
} For example, it is possible to perform automated serial sectioning of
} resin-embedded tissues and cells using a scanning EM. The development of
} an in-microscope ultramicrotome by Denk and Horstmann (PLoS Biol. 2004
} Nov;2(11):e329. Epub 2004 Oct 19. Serial block-face scanning electron
} microscopy to reconstruct three-dimensional tissue nanostructure) will
} greatly assist anyone wanting to produce 3-D reconstructions through large
} volumes of tissue. Basically, the embedded ultramicrotome slices a thin
} slice from the resin block and the SEM automatically images the block
} surface using the back-scatter electron detector. This attachment is on
} sale commercially as the 3-View system.
}
} A technique called “array tomography” also makes use of resin embedded
} sections, but in this instance, the sections are first mounted on glass
} slides so that they can be imaged by confocal microscopy first (search
} PubMed using “array tomography smith” for details). Array tomography makes
} it possible to label thin sections with antibodies and fluorescent dyes
} and then overlay SEM images of the same regions to produce reference space
} data (instead of the black space that the confocal presents).
}
} Dual beam microscopes offer an alternative approach to “sectioning”
} biological material while imaging it at high resolution. Imagine being
} able to slice through a cell and image the Golgi complex, “look for”
} endocytosed markers inside cells in real time, or look for intracellular
} viral particles.
}
} Paul Walther in Germany (Höhn et al. Histochem Cell Biol. 2010 Nov 27.
} Preparation of cryofixed cells for improved 3D ultrastructure with
} scanning transmission electron tomography) has been quietly pioneering the
} use of cryo-specimen stages on SEMs to image the inside of cells. One of
} his approaches is to cryo-plane specimens (cryo-section with a diamond
} knife) and image the cut surfaces in the SEM, and collecting images as
} frozen water gradually sublimates in the high vacuum of the microscope.
} Being able to prepare specimens for SEM using high pressure freezing and
} freeze substitution is an advantage for imaging with only minor
} ultrastructural changes.
}
} Some attempts have been made to image colloidal gold particles in the SEM
} and I think this approach will expand once the methods have been worked
} out. Imagine watching the aggregation of gold particles on cell surfaces,
} and their internalization into coated pits. Similarly, it should be
} possible to image isolated organelles and examine vesicle fusion, or
} attachment to actin filaments or microtubules.
}
}
} Modern SEMs offer a wide range of options. Variable pressure machines make
} it possible to image biological material without metal coating, high
} resolution machines offer the ability to “look at” high resolution
} structures on cell surfaces as well as intracellular membranes. Dual beam
} machines and embedded tools (such as ultramicrotomes or AFMs) give us ways
} of manipulating specimens that were previously not possible (or not easy).
} Combining light microscopy with the scanning EM (in situ hybridization or
} immunolabeling – e.g. Schaudinn et al. J Microsc. 2009 Aug;235(2):124-7.
} Imaging of endodontic biofilms by combined microscopy (FISH/cLSM - SEM)
} reveals structural detail not available if only one of these imaging modes
} is used.
}
} In conclusion, although there are many technical reasons why a biology
} department should have a scanning EM available for use (in addition to at
} least one high-end TEM), perhaps the most compelling reason to have an SEM
} available is that post-docs and graduate students will always find
} surprising new ways of imaging cells and tissues.
}
} Happy Holidays,
}
} Paul Webster
} House Ear Institute
} 2100 W 3rd St.
} Los Angeles
} CA 90057
} (213) 273 8026
}
}
} Bard,

One excellent reason is that a modern FE-SEM can reveal the true state of
cellular extensions - microvilli, pseudopods of various kinds, nanotubes
connecting cells and so on. These are very difficult to study in a TEM, and
many of their important features are sub-resolution in the light microscope.
These are important functional features of cells.

An even better reason is immunolabelling of cell surface features. Receptor
distributions, numbers, associations between and within receptor types and
so on. These are very difficult or worse to study in a TEM, since one is
looking at a 60-100 nm section through the plasmalemma, so receptors are
easily missed. In a SEM, the entire cell surface can be imaged, and all of
the receptors labelled and counted, or whatever.
For a quick example, check the U. Wisconsin BBPIC web page on
immunolabelling:
http://www.ansci.wisc.edu/facstaff/Faculty/pages/albrecht/albrecht_web/Programs/microscopy/colloid.html
There is an image gallery at the bottom of the page.
Further, by using colloidal metals of different shapes, one can do
multiple-labelling for SEM, and if their atomic number is sufficiently
different, potentially the labels can be distinguished by a good backscatter
detector.

Phil

Philip Oshel
Microscopy Facility Supervisor
Department of Biology
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576




-----Original Message-----
X-from: bard.smedsrod-at-uit.no [mailto:bard.smedsrod-at-uit.no]
Sent: Tue 10/12/21 17:02
To: Oshel, Philip Eugene

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at
http://microscopy.com/MicroscopyListserver/MLFormMail.html
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Email: bard.smedsrod-at-uit.no
Name: Bard Smedsrod

Organization: University of Tromso, Norway

Title-Subject: [Filtered] Need convincing arguments motivating purchase of
SEM

Message: Our Medical faculty has a well equipped EM platform, with
highly skilled staff. But our SEM instrument is really old. This
means it can only produce low magnification, low resolution images of
samples. We have struggled with this "pre-historic" SEM instrument
for years, and are trying to convince our Faculty that our
researchers deserve a new high quality, high resolution SEM
instrument equipped with the latest technology. Our problem now is
that our researchers have quit using SEM because the old instrument
has either been out of work or produced images that are not good
enough to publish. After many years with access to only a mediocre
SEM instrument our researchers have forgotten about the potential
benefits of using SEM, and the motivation among our research staff to
use and therefore demand this methodology is litterally gone.
Research at our Faculty that might benefit from including SEM
technology include: Physiology (cardiophysiology), Vascular biology
(special emphasis on liver sinusoids), Microbiology (microfilm
studies, and gene transfer among bacteria), Virology, Tumor biology
(both molecular and cell/organ based), Pharmacology, Pharmacy, Cell
biology (both molecular and "classical" cell biology), Immunology (in
particular T-cell mediated responses). I am convinced that some of
these research groups would benefit greatly from including a modern,
high-end SEM instrument in their studies. My task as head of the EM
platform is to awaken and motivate potential users (who have
forgotten the potential benefits of using SEM after all those years
with access only to a museum type of SEM) to understand that they may
benefit from using SEM. This is why I contact the Microscopy
ListServer. My hope is that some of you can give me some striking and
convincing examples showing how a high quality SEM may add valuable
dimensions to scientific studies in the biomedical fields mentioned
above. I should be really thankful to get a feedback including
concrete examples (references to high quality papers)!
leaving
no doubt that a high-end SEM enables high quality research. With
such arguments I think I might motivate our researchers to exert
sufficient pressure on our Faculty to purchase a high end SEM.
Regards,
Bard Smedsrod
Professor and Head of Vascular Biology Research Group
Scientific head of Laboratory of Electron Microscopy
Department of Medical Biology
University of Tromso
NO-9037 Tromso
NORWAY
tel.: +47-77644687 / +47-99599463





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From: FMonson-at-wcupa.edu
Date: Fri, 24 Dec 2010 08:21:00 -0600
Subject: [Microscopy] Re: Iodine stain

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Lugol's Iodine will do.
URL: http://www.amazon.com/J-Crows-Lugols-Iodine-2-Liquid/dp/B001AEFM9Y

Gurr's Lugol

1 g Iodine
2 g KI
mix the two dry components together (you will see an interesting phenomenon: the KI will turn brown. I can't remember the "Why")
50 or 100ml distilled water.

NOTE: iodine itself is not very soluble in H2O.

Cheers,

Fred Monson
http://cmirt.wcupa.edu

________________________________________
X-from: germpore-at-sonic.net [germpore-at-sonic.net]
Sent: Thursday, December 23, 2010 8:27 PM
To: Monson, Frederick

sawyert-at-science.oregonstate.edu wrote:

} Name: Teresa Sawyer
}
} Organization: Oregon State University
}
} Title-Subject: [Filtered] Iodine stain
}
} Message: Our Plant Anatomy class needs a stain for starch detection.
} IKI (potassium iodide + iodine) has been used in the past but they
} are having difficulty finding this stain. Any help location this
} stain or an alternate stain would be great.

Hi –

I'm unclear as to why you would be having difficulty obtaining IKI.
The standard IKI solution is Lugol's iodine, and not only is that
available through sources like Carolina and VWR, I've even it sold
over-the-counter through "green" pharmacies, and over Amazon.com.
Lugol's is good enough for simple starch detection and there's no
reason you should have difficulty obtaining either it, or stock iodine
and potassium iodide to make up your own. It would surprise me, in
fact, if your department's stockroom didn't have these already.

The difficult-to-find iodine solution you might be thinking of is
Melzer's, in which the IKI is in a chloral hydrate/aqueous solution
rather than a straight aqueous solution. The chloral hydrate gives the
solution additional clearing properties which are useful with heavily
pigmented tissue. However, due to the Schedule IV status of chloral
hydrate and the DEA's stepped up enforcement of licensing of such
compounds, it's difficult to get the ingredients to make unless one is
willing to go through DEA paperwork. I posted a question about
possible Melzer's substitutes to this list only a month ago, because
I've been up against the same problem.

However, being a bit familiar with your department (Botany & Plant
Pathology -at-OSU, right?), I know there are a number of mycologists in
that department (such as Joey Spatafora) who use Melzer's as a
standard tool. If Melzer's is in fact what you need, I would talk to
those people about where to obtain it.

Peter



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From: FMonson-at-wcupa.edu
Date: Fri, 24 Dec 2010 08:26:52 -0600
Subject: [Microscopy] Re: Iodine for starch

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Lugol's Iodine will do.
URL: http://www.amazon.com/J-Crows-Lugols-Iodine-2-Liquid/dp/B001AEFM9Y

Gurr's Lugol's Iodine Recipe

1 g Iodine
2 g KI
mix the two dry components together (you will see an interesting phenomenon: the KI will turn brown. I can't remember the "Why")
50 or 100ml distilled water.
NOTE: one can double the KI and I and halve the water.
NOTE: iodine itself is not very soluble in H2O.

Question: Why was this solution used in histologic staining methodoloy?. Other than for starch?

Cheers,

Fred Monson
http://cmirt.wcupa.edu

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From: zaluzec-at-aaem.amc.anl.gov
Date: Fri, 24 Dec 2010 10:37:29 -0600
Subject: [Microscopy] In memory: David Cockayne

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Colleagues

I have just learned the sad news that one of our colleagues David
Cockayne (FRS, Emeritus Professor Oxford)
has passed away. He died peaceful at home with family on Wednesday.

I, as well as many of you, will all miss his fireindship,
insightfulness and collegial
role in our community. On behalf of all of us, I will extend our
condolences to his family and friends.

Nestor

--
===========================================
Dr. Nestor J. Zaluzec
Argonne National Laboratory
Electron Microscopy Center
Materials Science Division/Bldg 212
9700 S. Cass Ave
Argonne, Illinois 60439 USA

Tel: 530-NES-TORZ (530-637-8679), Fax: 630-252-4798

iChat: Zaluzec-at-AIM.com
Skype: Zaluzec-at-ANL
Polycom: 146.139.72.119
TPM: http://tpm.amc.anl.gov

Email: Zaluzec-at-aaem.amc.anl.gov

Senior Scientist - Argonne National Laboratory
Fellow of the Microscopy Society of America
Senior Fellow the Computational Institute - University of Chicago
E.P. Wigner Fellow - Oak Ridge National Laboratory
President-Elect Microscopy Society of America






===========================================
TPMLab: http://tpm.amc.anl.gov
MMSite: http://www.amc.anl.gov
===========================================

The box said ...
"This program requires Win 95/98/NT or better..."
So I bought a Mac !

===========================================

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From: oshel1pe-at-cmich.edu
Date: Fri, 24 Dec 2010 11:48:27 -0600
Subject: [Microscopy] viaWWW: Need convincing arguments motivating purchase

Contents Retrieved from Microscopy Listserver Archives
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Yorgos,

I have to disagree with your statement about how much magnification is useful when using a SEM.
I have often found 20,000X and more helpful when doing ultrastructure of pollen and arthropod sensory setae (for 2 quick examples), and 20-100,000X necessary for doing immunolabelling of cell surface receptors. Plus, viri really need 90,000-400,000X, especially for some of the tiny bacteriophages.
These high mags also require a high-resolution instrument.
But I do agree there is a lot of excellent work done, and to be done, in an "old style" tungsten emitter SEM at a few hundred to a few thousand times magnification. (Like we have.)

Phil

Philip Oshel
Microscopy Facility Supervisor
Department of Biology
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576




-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: Fri 10/12/24 04:43
To: Oshel, Philip Eugene

Dear Brad



In biological research, SEM is an indispensable tool, primary or adjunctive,
for any morphological / ultrastructural study. As pointed out by other
colleagues, it can give you a unique sense of the 3-D structure of the cell
surface and the architecture of the tissues. In the field of human
reproduction I am familiar with, SEM is the only means to:

-detect the pinopodes, characteristic membrane projections of the
endometrial epithelium related to receptivity

-detect salpingeal or squamous metaplasias and their extend in the
endometrial epithelium

-to resolve the 3-D structure of the zona pellucida of the oocyte

-assess the extend of cell to cell communication in cell cultures or the
cell degeneration and apoptosis in the same systems

-to detect sperm abnormalities and quantify them

-and many others, for instance the changes of the endothelium in small
arteries due to hypertension



I have published more than 30 original and review articles based partially
or entirely in SEM.(can have a look at my web site www.aim.cat -click
publications). An additional advantage is that, because SEM images are
spectacular and easily appreciated, you have a good chance to make the cover
of the journal you publish. To my mind, SEM is a necessary adjunction to any
TEM study, if you want to have the full picture of the object you study.



However, I don't agree with the need to buy a very powerful instrument for
biological applications, not even a new one. As you know, in viewing
biological specimens under SEM, you will never go above X3K, maximum X5-10K
because simply there is nothing to see further on. And SEM images in the
field of biology are not different to what was taken 40 years ago. Clearly,
it is the preparation of the soft tissue that is crucial for the quality of
the image. You mention that your old instrument does not produce decent
images; this is maybe due to a technical problem which you can fix.



I don't think that a very modern instrument will really attract more
investigators. It can work the other way around, that if you have a super
scope and still the investigators see poor images due to their poor
preparations they will permanently switch away from SEM. Probably the best
idea is to get a good old instrument, maybe from between 80- 90ies, one
instrument that was widely used and there are still many spare parts around.
In this way you will avoid the complicated electronics and software
additions that modern scopes have and that makes you depending entirely from
the company which will inevitably screw you, (as it happened to me) and in a
few years you will have to scrap the scope because no more software /
hardware support. In an old good instrument you can add an external scan
generator (I am very happy with DISS5 from the German firm
"pointelectronic") and digitize the old scope with little money and be able
to get fantastic pictures of 50 or 80 mega pixels.



I hope that my comments will be helpful to you. In the topic you discuss, as
happens everywhere, it is more the man than the machine that makes the
difference.



Happy holidays to all of you in this lovely list

yorgos



Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
eikonika-at-otenet.gr
www.aim.cat
*********************************







}
} This is an interesting request in that many years ago I would have replied
} with the retort that biologists have no use for scanning electron
} microscopy. I was brought up with the idea that all useful information can
} be obtained using thin sections and a transmission EM.
}
} However, I run a shared imaging facility and was instructed to get a
} scanning EM as part of the initial set-up. We had money available for a
} good quality TEM and associated specimen preparation equipment (especially
} a cryo-ultramicrotome) so it did not bother me that we would have a
} scanning EM sitting unused. How wrong I was.
}
} First, let me say that my scanning EM is now 11 years old and has more use
} than the TEM sitting in the next room. There are many reasons for why this
} is, but the reason most often quoted by my users is that it is easy to
} prepare specimens for the machine, and also the most easy to use. Having
} trained how to operate confocal microscopes I can see how the ease of use
} compares.
}
} We can easily train users to operate the microscope because it is fully
} controlled by software and mouse clicks. Every year we host one or two
} summer students who are often in their final year of high school, and they
} take to the SEM rapidly. within a week they are able to train others and
} their enthusiasm turns them into effective ambassadors for what we do.
}
} The samples we have examined in the scanning EM have been multiple and
} varied. We first used it to look at the organ of Corti in the mammalian
} inner ear, a particularly well organized structure that offers much
} information on mammalian hearing. As I work in a a hearing research
} institute it is important that we are able to look at the inner ear
} structures. One particular structure that is difficult to image in the TEM
} is the thin protein bridge between the stereocillia (sticking out of the
} top of the sensory hair cells). Being able to obtain the resolution that
} reveals the individual proteins on this bridge is amazing.
}
} We have had users who image frog ears, zebrafish, individual neurons
} growing in culture, extracted neurons with myosin-decorated actin
} filaments, cultured cells, vibratome sections, dissected tissues,
} nanoparticles, synthetic organs formed on polymer scaffolds, and much
} more.
}
} Another use of the SEM that I had not initially foreseen was being able to
} image bacteria attaching to mammalian cell surfaces. Revealing how the
} cells respond to the bacteria was fascinating, as was the process by which
} the bacteria are engulfed by the cells. Of course it didn't take us long
} to move on to imaging activated and non-activated macrophages. These
} images drew in graduate students who wanted to learn how to obtain similar
} images.
}
} Our study of extracellular bacteria took us in a slightly different
} direction and resulted in us studying bacterial biofilms. The SEM was an
} essential part of us revealing complex 3-D structures formed by
} aggregations of extracellular bacterial proteins. The biofilm structures
} that we are imaging point to the possibility that bacteria may have an
} existence that is more than as individual cells.
}
} Our 11-year old SEM is now looking very old and we are in the process of
} discussing how we could replace it. First we will have to find the money,
} but then we will have to decide in which direction we would like to go if
} we do buy a new machine.
}
} We have been discussing the different microscope options available and
} have been talking with the microscope manufacturers. From these
} discussions it is clear that we need to upgrade as soon as possible. The
} options available for biologists are amazingly varied and useful.
}
} Talk with SEM suppliers and you will find an impressive amount of
} technological development. Modern scanning EMs offer different
} combinations of high or low vacuum, high resolution and an impressive
} amount of novel hardware and software. How these tools are used seems to
} be limited only by the imagination of the operator,
}
} For example, it is possible to perform automated serial sectioning of
} resin-embedded tissues and cells using a scanning EM. The development of
} an in-microscope ultramicrotome by Denk and Horstmann (PLoS Biol. 2004
} Nov;2(11):e329. Epub 2004 Oct 19. Serial block-face scanning electron
} microscopy to reconstruct three-dimensional tissue nanostructure) will
} greatly assist anyone wanting to produce 3-D reconstructions through large
} volumes of tissue. Basically, the embedded ultramicrotome slices a thin
} slice from the resin block and the SEM automatically images the block
} surface using the back-scatter electron detector. This attachment is on
} sale commercially as the 3-View system.
}
} A technique called "array tomography" also makes use of resin embedded
} sections, but in this instance, the sections are first mounted on glass
} slides so that they can be imaged by confocal microscopy first (search
} PubMed using "array tomography smith" for details). Array tomography makes
} it possible to label thin sections with antibodies and fluorescent dyes
} and then overlay SEM images of the same regions to produce reference space
} data (instead of the black space that the confocal presents).
}
} Dual beam microscopes offer an alternative approach to "sectioning"
} biological material while imaging it at high resolution. Imagine being
} able to slice through a cell and image the Golgi complex, "look for"
} endocytosed markers inside cells in real time, or look for intracellular
} viral particles.
}
} Paul Walther in Germany (Höhn et al. Histochem Cell Biol. 2010 Nov 27.
} Preparation of cryofixed cells for improved 3D ultrastructure with
} scanning transmission electron tomography) has been quietly pioneering the
} use of cryo-specimen stages on SEMs to image the inside of cells. One of
} his approaches is to cryo-plane specimens (cryo-section with a diamond
} knife) and image the cut surfaces in the SEM, and collecting images as
} frozen water gradually sublimates in the high vacuum of the microscope.
} Being able to prepare specimens for SEM using high pressure freezing and
} freeze substitution is an advantage for imaging with only minor
} ultrastructural changes.
}
} Some attempts have been made to image colloidal gold particles in the SEM
} and I think this approach will expand once the methods have been worked
} out. Imagine watching the aggregation of gold particles on cell surfaces,
} and their internalization into coated pits. Similarly, it should be
} possible to image isolated organelles and examine vesicle fusion, or
} attachment to actin filaments or microtubules.
}
}
} Modern SEMs offer a wide range of options. Variable pressure machines make
} it possible to image biological material without metal coating, high
} resolution machines offer the ability to "look at" high resolution
} structures on cell surfaces as well as intracellular membranes. Dual beam
} machines and embedded tools (such as ultramicrotomes or AFMs) give us ways
} of manipulating specimens that were previously not possible (or not easy).
} Combining light microscopy with the scanning EM (in situ hybridization or
} immunolabeling - e.g. Schaudinn et al. J Microsc. 2009 Aug;235(2):124-7.
} Imaging of endodontic biofilms by combined microscopy (FISH/cLSM - SEM)
} reveals structural detail not available if only one of these imaging modes
} is used.
}
} In conclusion, although there are many technical reasons why a biology
} department should have a scanning EM available for use (in addition to at
} least one high-end TEM), perhaps the most compelling reason to have an SEM
} available is that post-docs and graduate students will always find
} surprising new ways of imaging cells and tissues.
}
} Happy Holidays,
}
} Paul Webster
} House Ear Institute
} 2100 W 3rd St.
} Los Angeles
} CA 90057
} (213) 273 8026
}
}
} Bard,

One excellent reason is that a modern FE-SEM can reveal the true state of
cellular extensions - microvilli, pseudopods of various kinds, nanotubes
connecting cells and so on. These are very difficult to study in a TEM, and
many of their important features are sub-resolution in the light microscope.
These are important functional features of cells.

An even better reason is immunolabelling of cell surface features. Receptor
distributions, numbers, associations between and within receptor types and
so on. These are very difficult or worse to study in a TEM, since one is
looking at a 60-100 nm section through the plasmalemma, so receptors are
easily missed. In a SEM, the entire cell surface can be imaged, and all of
the receptors labelled and counted, or whatever.
For a quick example, check the U. Wisconsin BBPIC web page on
immunolabelling:
http://www.ansci.wisc.edu/facstaff/Faculty/pages/albrecht/albrecht_web/Programs/microscopy/colloid.html
There is an image gallery at the bottom of the page.
Further, by using colloidal metals of different shapes, one can do
multiple-labelling for SEM, and if their atomic number is sufficiently
different, potentially the labels can be distinguished by a good backscatter
detector.

Phil

Philip Oshel
Microscopy Facility Supervisor
Department of Biology
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576




-----Original Message-----
X-from: bard.smedsrod-at-uit.no [mailto:bard.smedsrod-at-uit.no]
Sent: Tue 10/12/21 17:02
To: Oshel, Philip Eugene

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Email: bard.smedsrod-at-uit.no
Name: Bard Smedsrod

Organization: University of Tromso, Norway

Title-Subject: [Filtered] Need convincing arguments motivating purchase of
SEM

Message: Our Medical faculty has a well equipped EM platform, with
highly skilled staff. But our SEM instrument is really old. This
means it can only produce low magnification, low resolution images of
samples. We have struggled with this "pre-historic" SEM instrument
for years, and are trying to convince our Faculty that our
researchers deserve a new high quality, high resolution SEM
instrument equipped with the latest technology. Our problem now is
that our researchers have quit using SEM because the old instrument
has either been out of work or produced images that are not good
enough to publish. After many years with access to only a mediocre
SEM instrument our researchers have forgotten about the potential
benefits of using SEM, and the motivation among our research staff to
use and therefore demand this methodology is litterally gone.
Research at our Faculty that might benefit from including SEM
technology include: Physiology (cardiophysiology), Vascular biology
(special emphasis on liver sinusoids), Microbiology (microfilm
studies, and gene transfer among bacteria), Virology, Tumor biology
(both molecular and cell/organ based), Pharmacology, Pharmacy, Cell
biology (both molecular and "classical" cell biology), Immunology (in
particular T-cell mediated responses). I am convinced that some of
these research groups would benefit greatly from including a modern,
high-end SEM instrument in their studies. My task as head of the EM
platform is to awaken and motivate potential users (who have
forgotten the potential benefits of using SEM after all those years
with access only to a museum type of SEM) to understand that they may
benefit from using SEM. This is why I contact the Microscopy
ListServer. My hope is that some of you can give me some striking and
convincing examples showing how a high quality SEM may add valuable
dimensions to scientific studies in the biomedical fields mentioned
above. I should be really thankful to get a feedback including
concrete examples (references to high quality papers)!
leaving
no doubt that a high-end SEM enables high quality research. With
such arguments I think I might motivate our researchers to exert
sufficient pressure on our Faculty to purchase a high end SEM.
Regards,
Bard Smedsrod
Professor and Head of Vascular Biology Research Group
Scientific head of Laboratory of Electron Microscopy
Department of Medical Biology
University of Tromso
NO-9037 Tromso
NORWAY
tel.: +47-77644687 / +47-99599463





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From: connellyps-at-nhlbi.nih.gov
Date: Wed, 29 Dec 2010 09:40:16 -0600
Subject: [Microscopy] TEM need help with staining

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Lawrence,

I studied melanosomes in melanoma cells many years ago and never had a stain
problem.

When staining sections that are cut from samples that were block stained I
have not used a concentration higher than 2% while others use up to 4%. Are
you sure you need 8% UA? My standard section stain is 1% UA having come from
a stock of 2% UA in water that I dilute with 100% Ethanol - 1 drop of each
on parafilm/grid. Staining time is 10 minutes, in a covered dish to keep
the light out, before washing. This is followed by lead staining.

I do know that UA can go bad especially if the crystals or solutions are
exposed to light so keep the UA dark except when making up the staining
solution and cover the bottle while stirring. The first thing I would do is
open a bottle of UA to make up new stain, a bottle that has been in its box
or can and not sitting beside the bottle on the shelf that you have been
using. You will need to put the bottle of UA back into its can to keep it
dark for future use.

I did not see the timing for the block stain that you used. Overnight
staining of the samples is not a bad idea. In the past I have used 2%UA in a
60 degree oven for the melanocytes (JCB 50:540-544,1971) but now have been
routinely leaving other samples in 1% UA in a refrigerator. Both have
worked well although I can not explain the two temperatures other than newer
references. If I need to hurry a sample along I'll block stain for 1 hour
at room temperature but this is usually not up to normal quality.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E ­ Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-mail.nih.gov {mailto:connellyps-at-mail.nih.gov}

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and can be freely shared
and reproduced.




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From: maimai_huang-at-seed.net.tw
Date: Fri, 31 Dec 2010 10:40:08 -0600
Subject: [Microscopy] viaWWW: About C contamination in Cs-corrected TEM

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Email: maimai_huang-at-seed.net.tw
Name: Michael Huang

Organization: National Cheng Kung University

Title-Subject: [Filtered] About C contamination in Cs-corrected TEM

Message: Dear all seasoned TEM users:

My name is Michael Huang from Taiwan and I would like to consult you
all some questions related to carbon contamination.

Recently we luckily got our new plasma cleaner. However, we still met
troublous carbon contamination problem during the experiment
(Cs-corrected TEM). The contamination seemed to be very serious while
conducting S.I. particularly with very fine scanning mesh setup. The
contamination occurring at vacuum / sample interface is most
difficult to prohibited even longer cleaning time applied. I swore I
saw the growth of contamination from interface toward vacuum space,
which is almost unbelievable!!

Our sample is nanoparticles collected on lacy grid with alcohol
solution and then heated for 1 hour at 80C.

Have anyone met such ridiculous problem while operating TEM?

Thanks for your help and happy New Year.

BR

Michael


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From: maimai_huang-at-seed.net.tw
Date: Fri, 31 Dec 2010 10:40:31 -0600
Subject: [Microscopy] viaWWW: About bug (maybe..) in new version DigitalMicrograph

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Email: maimai_huang-at-seed.net.tw
Name: Michael Huang

Organization: National Cheng Kung University

Title-Subject: [Filtered] About bug (maybe..) in new version DigitalMicrograph

Message: Dear all seasoned TEM users:

My name is Michael Huang from Taiwan and I would
like to consult you all some questions about
DigitalMicrograph (DM).

Recently I installed new version DM (1.7 or 1.8).
Initially I was excited to make use of such
fashionable version of various new functions, but
later I felt disappointed while processing my low
loss spectrum. It seems that the derived results
are slightly different from the one obtained
previously with earlier v.1.4, and sometimes
problem emerged in doing ìremove plural
scatteringî, ìextract ZLPî and KKA. The spectrum
analysis functions of new version seem not as
reliable as v1.4.

Anyone observes this problem, and are there really some bugs in the software?

Thanks for your help and happy New Year.

BR

Michael


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