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From: l.tetley-at-bio.gla.ac.uk
Date: Tue, 1 Nov 2005 06:51:20 -0600
Subject: [Microscopy] 33rd Scottish Microscopy Symposium

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

33rd Scottish Microscopy Symposium


Wednesday 9th November 2005, Hunter Halls, University of Glasgow, Glasgow
G12 8QQ, Scotland, UK

Registration - with a 75% discount for students - is via

http://www.gla.ac.uk/ibls/II/em/SMG/smgnew.html

with a programme available at

http://www.gla.ac.uk/ibls/II/em/SMG/programme.pdf


plus a 25 company Trade Exhibition :

http://www.gla.ac.uk/departments/ibls/II/em/SMG/trade.html.


All welcome but registration forms should be FAXed to Will Maxwell at 0141
330 4299 in order to assess numbers for catering. This can be followed by
payment made on arrival.

Dr Laurence Tetley
Division of Infection & Immunity, IBLS,
Integrated Microscopy Facility
Joseph Black Building
University of Glasgow
Glasgow G12 8QQ

l.tetley-at-bio.gla.ac.uk
tel 0141 330 4431
FAX 0141 330 3516

Integrated Microscopy
Facility: http://www.gla.ac.uk/Acad/IBLS/II/em/mcb-em.htm
Cryo Microscopy Group: http://www.cryomicroscopygroup.org.uk
Royal Microscopical Society: http://www.rms.org.uk


Dr Laurence Tetley
Division of Infection & Immunity, IBLS,
Integrated Microscopy Facility
Joseph Black Building
University of Glasgow
Glasgow G12 8QQ

Tel 0141 330 4431
FAX 0141 330 3516

Integrated Microscopy Facility: http://www.gla.ac.uk/Acad/IBLS/II/em/
Royal Microscopical Society: http://www.rms.org.uk
Scottish Microscopy Group: http://www.gla.ac.uk/ibls/II/em/SMG/smgnew.html


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From: microscopytoday-at-tampabay.rr.com
Date: Tue, 1 Nov 2005 11:36:28 -0600
Subject: [Microscopy] November 2005 Microscopy Today Table of Contents

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

Listers,

Here is the November 2005 Microscopy Today table of contents. I will
close the subscription list for this issue on Monday, November 7, 2005.

Microscopists in North America and MSA members anywhere may have free
subscriptions. Anyone else may subscribe for US$35 per year (to
PARTIALLY cover postage). All subscriptions at
http://www.microscopy-today.com Thank you. Non-Qualified subscription
rates will increase to US$50 in 2006.

Ron Anderson, Editor
==================================

Very Cool Clathrin
Stephen W. Carmichael, Mayo Clinic

Metallography for the European Copper Age: Research on the Axe-Blade of
the Glacier-Mummy from the Ötztaler Alps in Tyrol
Gerhard O. Sperl, Institute for Historical Materials, Leoben, Austria

Microscopy and Microbes at Plum Island: Protecting America’s Livestock
Thomas G. Burrage, Plum Island Animal Disease Center, NY

Color Metallography
George F. Vander Voort, Buehler Ltd, Lake Bluff, Illinois

Characterization of Solids from Oilfield Emulsions
Richard W. Cloud,† Rebecca L. Ramsey,‡ Robert A. Pultz,‡ and Michael K.
Poindexter‡, † Nalco Company, Naperville, Illinois; ‡ Nalco Energy
Services, Sugar Land, Texas

Automated, Robotic Preparation of Vitrified Samples for 2D and 3D Cryo
Electron Microscopy
P. M. Frederik1 and M.H. Storms2; 1Univ. Maastricht, The Netherlands, 2.
FEI Company, Achtseweg Noord Eindhoven, The Netherlands

Ex-Situ “Auto Lift” Technique for TEM Sample Preparation
Garth “Brian” Cook, Micro Optics of Florida, Davie, Florida

Writing Nano-Scale Patterns on Insulators Using Variable Pressure
Electron-Beam Lithography
Floyd Miller and David Frey,* Lehigh University Bethlehem, PA and *Carl
Zeiss SMT Inc
.
Microscopy for Children
Caroline Schooley, MSA Project MICRO

Fostering LIMS Development Through Open Standards
Avrum Goodblatt, U. Penn School of Medicine, Philadelphia

50th Annivsary Celebrations of Atomic-Resolution Imaging
Thomas F. Kelly and Allan J. Melmed

Attaching Spheres to Cantilevers for Colloidal Probe Force Measurements:
A Simplified Technique
Yang Gan, University of Newcastle, Callaghan, NSW, Australia

Nissl: The Man, The Stain and The Substance
John A. Kiernan, The Univ. of Western Ontario London, Canada

Industry News
NetNotes
Index of Advertisers


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From: wgunning-at-meduohio.edu
Date: Wed, 2 Nov 2005 07:27:05 -0600
Subject: [Microscopy] viaWWW: MSA Award Nominations Reminder

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This Question was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html
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Email: wgunning-at-meduohio.edu
Name: WT Gunning

Organization: Medical University of Ohio

Title-Subject: [Filtered] MListserver: MSA Award Nominations Reminder

Question: Please note that MSA Awards nominations are due on December 15th, 2005.

2006 MICROSCOPY SOCIETY OF AMERICA AWARDS

The Distinguished Scientist Awards
(Physical & Biological Sciences)

The Burton Medal

The Outstanding Technologist Awards
(Physical & Biological Sciences)

The Mort Maser Distinguished Service Award

All MSA Members are encouraged to nominate candidates for these awards to recognize our eminent Scientific and Society leaders. Details available from the MSA Business Office.

Bostrom Corp.
230 East Ohio, Suite 400
Chicago, IL 60611
Toll-free: 1-800-538-3672
Tel: 312-644-1527
Fax: 312-644-8557
Email: BusinessOffice-at-microscopy.org

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From: camiller-at-anatomy.iupui.edu
Date: Wed, 2 Nov 2005 07:32:55 -0600
Subject: [Microscopy] viaWWW: Spring Experimental Biology 2006 Meeting

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Email: camiller-at-anatomy.iupui.edu
Name: Caroline Miller

Organization: Indiana University School of Medicine

Title-Subject: [Filtered] MListserver:

Question: We are approaching an abstract deadline for the Spring Experimental Biology 2006 Meeting in San Francisco April 1-5, 2006 (a national meeting with an attendance of about 15,000 people, see FASEBís website www.faseb.org). There is a platform session entitled ìElectron Microscopy as a 21st Century Toolî with a 15 minute talks. This platform session is entitled ìElectron Microscopy as a 21st Century Toolî and sponsored by the American Association of Anatomists-AAA and is being supported financially, in part, by the Indiana Microscopy Society and a request support from the MSA will be submitted. If you are interested in presenting some of your cutting edge, biological EM data, please consider submitting an abstract for the session. You will need to submit an abstract (the website URL for submission of an abstract is below). I will try to get sufficient funding so that all who present in this session can get reimbursed for the abstract fee as well as your registration and other associated expenses (hopefully) after the meeting. Make sure you submit for the AAA platform session entitled ìElectron Microscopy as a 21st Century Toolî.


Abstract submission deadline is November 7, 2005 . Thank you for considering submitting for this meeting



Abstract submission website (http://submissions.miracd.com/eb2006/login.asp)





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From: Willem.Wennekes-at-comcast.net
Date: Wed, 2 Nov 2005 07:44:18 -0600
Subject: [Microscopy] [Filtered] AskAMicroscopist: Aperture vs Current for EDX analysis

Contents Retrieved from Microscopy Listserver Archives
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This Question was submitted to Ask-A-Microscopist by (Willem.Wennekes-at-comcast.net)
from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, November 1, 2005 at 21:49:12
Remember to consider the Grade/Age of the student when considering the Question
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Email: Willem.Wennekes-at-comcast.net
Name: Willem Wennekes

Organization: UES

Education: Graduate College

Location: Dayton, OH, USA

Title: edx

Question: Hi, What would be the benefit of using a larger objective apperture instead of increasing the probe current in order to increase the beam current for edx analysis and x-ray mapping?

Cheers,
Willem

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From: David.R.Hull-at-nasa.gov
Date: Wed, 2 Nov 2005 10:21:26 -0600
Subject: [Microscopy] Re: [Filtered] AskAMicroscopist: Aperture vs Currentfor EDX analysis

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

}
} Email: Willem.Wennekes-at-comcast.net
} Name: Willem Wennekes
}
} Organization: UES
}
} Education: Graduate College
}
} Location: Dayton, OH, USA
}
} Title: edx
}
} Question: Hi, What would be the benefit of using a larger
} objective apperture instead of increasing the probe current in order
} to increase the beam current for edx analysis and x-ray mapping?
}
} Cheers,
} Willem

The benefit is that you keep your small aperture clean for imaging.

==============================Original Headers==============================
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From: kenconverse-at-qualityimages.biz
Date: Wed, 2 Nov 2005 11:01:30 -0600
Subject: [Microscopy] [Filtered] AskAMicroscopist: Aperture vs Current for

Contents Retrieved from Microscopy Listserver Archives
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Willem,
When you increase your probe size (demagnify the source less), the spray
electrons from each crossover tend to be captured closer to the beam path
thereby creating heavier contamination buildup nearer to the beam. This can
result in faster degradation of your imaging capabilities and the need for
more frequent cleaning of 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: Willem.Wennekes-at-comcast.net [mailto:Willem.Wennekes-at-comcast.net]
Sent: Wednesday, November 02, 2005 8:47 AM
To: kenconverse-at-qualityimages.biz

This Question was submitted to Ask-A-Microscopist by
(Willem.Wennekes-at-comcast.net)
from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on
Tuesday, November 1, 2005 at 21:49:12
Remember to consider the Grade/Age of the student when considering the
Question
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Email: Willem.Wennekes-at-comcast.net
Name: Willem Wennekes

Organization: UES

Education: Graduate College

Location: Dayton, OH, USA

Title: edx

Question: Hi, What would be the benefit of using a larger objective
apperture instead of increasing the probe current in order to increase the
beam current for edx analysis and x-ray mapping?

Cheers,
Willem

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

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==============================Original Headers==============================
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From: gary-at-gaugler.com
Date: Wed, 2 Nov 2005 11:34:55 -0600
Subject: [Microscopy] Re: AskAMicroscopist: Aperture vs Current for EDX

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

What are the options for increasing probe current? I.e., what
things can you do to increase it? yes, increasing the aperture
size will increase probe current. However, it increases probe
diameter. Another option is to increase KV. This will greatly
increase volumetric interacion such that more info comes from
deeper in the specimen. It is a constant trade-off problem.

Before changing anything above, be sure your specimen is at
the optimal analytical working distance (WD) for the EDS.
If you don't have that value, you can find it by experimentation.
Just start at a relatively long WD, check the counts per second
and then start reducing WD. Keep checking the cps at each
successive reduction in WD. The cps will start to increase and
then decrease. The WD at max cps is your analytical WD. Work
at this WD.

Now, back to aperture size and KV. I'd start with KV first.
Based on the highest Z you are looking at, you will need about
2X the eV of the line value you are collecting. Look at the M, K
and L series values and determine the necessary KV. E.g., Aluminum.
Z=13. No L-series lines. Ka=1.486KeV. Kb=1.553KeV. No N-series.
So, K-series are the only ones (BTW, Ka is the one used for calibration
along with Cu Ka at 8.040KeV.). So for Al, 5KV would probably
be a good value. If the specimen is bulk, then higher KV is OK
and would increase probe current and cps.

Take an extreme such as W (Z=74). Here, we have lines at all
shell series. However, not all of them are practical for normal
SEM. Ka=59.305KeV (can't use this since KV would need to be about
120KV). Same problem with Kb. Ma=1.774KeV. Sorting through the
other lines, La pops up at 8.394KeV. So 2X this would be 16KV.
So, 18KV-20KV would be appropriate. But at this KV, volumetric
interaction would be very high. High Z and high KV will result
in skyrocketing cps. So, smaller aperture would be necessary
to keep dead time down to {30% or so. If you want to quant,
this is probably the appropriate condition. However, for mapping,
if you do not have a volumetric interaction situation, and you
do not need high resolution, you can increase KV and aperture
size to increase cps. Keep in mind that most EDS systems need
cps to be such that dead time is {=35% or thereabouts. Increasing
cps with resulting higher DT simply throws away data since the
pulse processor cannot handle the high number of counts. Each
system has their own limit on this. Just look at DT to be sure
the processor is not overloaded.

Again, if the specimen is bulk versus very small, then increasing
aperture size will increase probe current and cps at the expense of
resolution. But for low mag, bulk specimens, not much of an issue.

Hope this helps.

gary g.







At 05:45 AM 11/2/2005, you wrote:

} November 1, 2005 at 21:49:12
} Remember to consider the Grade/Age of the student when considering
} the Question
} ---------------------------------------------------------------------------
} Please reply to both Willem.Wennekes-at-comcast.net as well as to
} the Microscopy Listserver
} ---------------------------------------------------------------------------
}
} Email: Willem.Wennekes-at-comcast.net
} Name: Willem Wennekes
}
} Organization: UES
}
} Education: Graduate College
}
} Location: Dayton, OH, USA
}
} Title: edx
}
} Question: Hi, What would be the benefit of using a larger
} objective apperture instead of increasing the probe current in order
} to increase the beam current for edx analysis and x-ray mapping?
}
} Cheers,
} Willem
}
} ---------------------------------------------------------------------------
}
} ==============================Original Headers==============================
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From: dhitrys-at-qimaging.com
Date: Wed, 2 Nov 2005 11:56:12 -0600
Subject: [Microscopy] Online Seminar: Selecting a Digital Camera for Light Microscopy

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

Announcing an interactive, web-based seminar:

===================================================
"Selecting a Digital Camera for Light Microscopy."
===================================================

Details are below.

Connection lines are limited, so reserve yours now. There is no charge to
participate in this on-line seminar.

For Wednesday, 9-November, 10:00 AM (New York time), pre-register at:
https://premconf.webex.com/premconf/j.php?ED=85888757&RG=1

For Thursday, 10-November, 11:00 AM (New York time, pre-register at:
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[ If any of the links have wrapped, please re-build it in your web browser's
address bar. The line begins with "https" and ends with "RG=1" ]


Details:
============

"Selecting a Digital Camera for Light Microscopy."
Presented by David Hitrys, QImaging Corporation

Attendee's will learn about digital cameras, how to interpret their
performance specifications, and will leave with enhanced skills for
critically analyzing a camera's suitability for various applications.

Outline of Topics:

-Understanding Digital Camera Parameters:
-Camera Sensitivity
-Noise Limitations
-Dynamic Range (Full-Well Capacity)
-Use of Gain
-Color Acquisition Options
-Note on Under- and Over-Sampling
-Matching Pixel Size to Optical Resolution
-Questions from the Audience

Presented by QImaging Corporation, makers of precision cameras for
microscopy, the information imparted will be broadly useful to anyone
striving to make the best camera choice for their imaging goals.

This seminar requires that attendees use a Java-enabled browser with a high
bandwidth connection. Audio is via toll-free telephone.

There is no charge to participate in this on-line seminar.

==============================Original Headers==============================
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From: beth-at-plantbio.uga.edu
Date: Wed, 2 Nov 2005 12:43:04 -0600
Subject: [Microscopy] support film - DuraSiN

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

Hi all,
Has anyone used the new TEM support films from DuraSiN?
I'd appreciate hearing your opinions on this product.

SINcerely,
Beth

**********************************************************************
Beth Richardson
EM Lab Coordinator
Plant Biology Department
University of Georgia
Athens, GA 30602-7271

Phone - (706) 542-1790 & FAX - (706) 542-1805
http://www.plantbio.uga.edu/emlab

"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)

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



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From: laable-at-solutia.com
Date: Wed, 2 Nov 2005 13:30:32 -0600
Subject: [Microscopy] SEM - repair service

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

Is anyone aware of an independent company who offers yearly service contracts
on LEO field emission SEMS. I would appreciate any info possible.

Thanks,
Lori Ables
laable-at-solutia.com


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From: schamber-at-aspexllc.com
Date: Wed, 2 Nov 2005 13:54:05 -0600
Subject: [Microscopy] AskAMicroscopist: Aperture vs Current for EDX

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

The statement that "increasing the aperture size ... increases probe
diameter" is not always true. The relationship between beam diameter
and beam current can be found in many references (e.g., equation 2.14 of
the 2nd edition of Goldstein). What this equation shows is that all
four of the terms that control beam diameter vary as the beam half-angle
(alpha) which is proportional to final aperture diameter. However, two
of the terms involve alpha in the numerator, and two in the denominator.
Thus, for any given probe current (Ip) there is an optimum aperture
size that gives the minimum beam diameter. Either a smaller or larger
aperture than called for by the minimum condition will make the beam
diameter larger.

Thus one very good reason for sometimes using a larger aperture size to
increase beam current rather than just cranking up the Spot control
(less demagnification) is to better match the condition for minimum spot
size .

Fred Schamber
ASPEX Corp.

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
} ----------------------------------------------------------------------------
}
} What are the options for increasing probe current? I.e., what
} things can you do to increase it? yes, increasing the aperture
} size will increase probe current. However, it increases probe
} diameter. Another option is to increase KV. This will greatly
} increase volumetric interacion such that more info comes from
} deeper in the specimen. It is a constant trade-off problem.
}
} Before changing anything above, be sure your specimen is at
} the optimal analytical working distance (WD) for the EDS.
} If you don't have that value, you can find it by experimentation.
} Just start at a relatively long WD, check the counts per second
} and then start reducing WD. Keep checking the cps at each
} successive reduction in WD. The cps will start to increase and
} then decrease. The WD at max cps is your analytical WD. Work
} at this WD.
}
} Now, back to aperture size and KV. I'd start with KV first.
} Based on the highest Z you are looking at, you will need about
} 2X the eV of the line value you are collecting. Look at the M, K
} and L series values and determine the necessary KV. E.g., Aluminum.
} Z=13. No L-series lines. Ka=1.486KeV. Kb=1.553KeV. No N-series.
} So, K-series are the only ones (BTW, Ka is the one used for calibration
} along with Cu Ka at 8.040KeV.). So for Al, 5KV would probably
} be a good value. If the specimen is bulk, then higher KV is OK
} and would increase probe current and cps.
}
} Take an extreme such as W (Z=74). Here, we have lines at all
} shell series. However, not all of them are practical for normal
} SEM. Ka=59.305KeV (can't use this since KV would need to be about
} 120KV). Same problem with Kb. Ma=1.774KeV. Sorting through the
} other lines, La pops up at 8.394KeV. So 2X this would be 16KV.
} So, 18KV-20KV would be appropriate. But at this KV, volumetric
} interaction would be very high. High Z and high KV will result
} in skyrocketing cps. So, smaller aperture would be necessary
} to keep dead time down to {30% or so. If you want to quant,
} this is probably the appropriate condition. However, for mapping,
} if you do not have a volumetric interaction situation, and you
} do not need high resolution, you can increase KV and aperture
} size to increase cps. Keep in mind that most EDS systems need
} cps to be such that dead time is {=35% or thereabouts. Increasing
} cps with resulting higher DT simply throws away data since the
} pulse processor cannot handle the high number of counts. Each
} system has their own limit on this. Just look at DT to be sure
} the processor is not overloaded.
}
} Again, if the specimen is bulk versus very small, then increasing
} aperture size will increase probe current and cps at the expense of
} resolution. But for low mag, bulk specimens, not much of an issue.
}
} Hope this helps.
}
} gary g.
}
}
}
}
}
}
}
} At 05:45 AM 11/2/2005, you wrote:
}
}
} } November 1, 2005 at 21:49:12
} } Remember to consider the Grade/Age of the student when considering
} } the Question
} } ---------------------------------------------------------------------------
} } Please reply to both Willem.Wennekes-at-comcast.net as well as to
} } the Microscopy Listserver
} } ---------------------------------------------------------------------------
} }
} } Email: Willem.Wennekes-at-comcast.net
} } Name: Willem Wennekes
} }
} } Organization: UES
} }
} } Education: Graduate College
} }
} } Location: Dayton, OH, USA
} }
} } Title: edx
} }
} } Question: Hi, What would be the benefit of using a larger
} } objective apperture instead of increasing the probe current in order
} } to increase the beam current for edx analysis and x-ray mapping?
} }
} } Cheers,
} } Willem
} }
} } ---------------------------------------------------------------------------
} }
} } ==============================Original Headers==============================
} } 9, 12 -- From zaluzec-at-ultra5.microscopy.com Wed Nov 2 07:44:18 2005
} } 9, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22])
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} } 9, 12 -- Subject: [Filtered] AskAMicroscopist: Aperture vs Current
} } for EDX analysis
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}
}
} ==============================Original Headers==============================
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} 16, 25 -- Subject: Re: [Microscopy] AskAMicroscopist: Aperture vs Current for EDX
} 16, 25 -- analysis
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==============================Original Headers==============================
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4, 25 -- From: Frederick Schamber {schamber-at-aspexllc.com}
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From: CGoldsmith-at-cdc.gov
Date: Wed, 2 Nov 2005 17:53:05 -0600
Subject: [Microscopy] viaWWW: Used FEI EM410-LS is available

Contents Retrieved from Microscopy Listserver Archives
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This Question was submitted to the Microscopy Listserver
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Email: CGoldsmith-at-cdc.gov
Name: Cynthia Goldsmith

Organization: Centers for Disease Control and Prevention (CDC)

Title-Subject: [Filtered] MListserver:Used FEI EM410-LS is available

Question: The Centers for Disease Control and Prevention (CDC) has a used FEI EM410-LS that is available for transfer to a government facility (federal, state, or local) or an educational institution (university, college, high school, etc.). Microscope is 21 years old, and in good condition. Please contact me off-line at CGoldsmith-at-cdc.gov for more information.


Cynthia S. Goldsmith, M.S.
Infectious Disease Pathology Activity
Centers for Disease Control and Prevention (CDC)
Phone: (404)639-3306
Fax: (404)639-1377



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From: gary-at-gaugler.com
Date: Wed, 2 Nov 2005 18:16:54 -0600
Subject: [Microscopy] Re: AskAMicroscopist: Aperture vs Current for EDX

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

This follows published info. However, the question was
about aperture size. The other variable of condenser
squeeze or relaxation is something else to throw into the
pot. We don't know if his system has a condenser control.
Probably so.

He was looking for increase in probe current. I figure
that it is safe to say that everything else being equal,
increasing the aperture size will increase probe current.
That is really all he wants to do/know--notwithstanding all
the other nuances. He just wants to know about a larger
final aperture size.

gary g.



At 11:55 AM 11/2/2005, you wrote:



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From: beth-at-plantbio.uga.edu
Date: Thu, 3 Nov 2005 09:31:16 -0600
Subject: [Microscopy] silicon nitride support film grids

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

Hi again,
I've been asked to rephrase my question about silicon nitride membrane
window grids.
If you use either the SPI silicon nitride membrane window grids or the
EMS DuraSiN silicon nitride membrane window grids I'd like to hear your
opinion of them.
Thanks,
Beth

**********************************************************************
Beth Richardson
EM Lab Coordinator
Plant Biology Department
University of Georgia
Athens, GA 30602-7271

Phone - (706) 542-1790 & FAX - (706) 542-1805
http://www.plantbio.uga.edu/emlab

"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)

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



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From: schamber-at-aspexllc.com
Date: Thu, 3 Nov 2005 11:16:45 -0600
Subject: [Microscopy] AskAMicroscopist: Aperture vs Current for EDX

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

I think there are two issues here. One is the "physics" question (what
does theory say? ) and the other a practical question (what happens in
an actual instrument operating in a normal way?).

First the physics. Here the ultimate limit is diffraction. Regardless
of the source type, a too-small aperture will cause a diffraction
spreading of the beam. The simplest way to understand this is as a
manifestation of the Heisenberg uncertainty principle -- when the
lateral location of the electron is too tightly constrained by the
aperture, then the lateral uncertainty in the momentum increases and
there is broadening. So if one were operating a SEM in a
difraction-limited mode, increasing the aperture size would be the most
sensible way to increase the beam current.

Most thermionic (not FE) SEMs are not operated anywhere near the
diffraction limit. Usually the dominant limit to resolution is
spherical aberration and that term varies as aperture size to the third
power. The simple quadrature approximation shown as equation 2.14 in
Goldstein (2nd Ed) is generally regarded as valid for thermionic SEMs,
but is not valid for any instrument operating near the diffraction
limit. Here one must use much more complex wave-function calculations
-- it's been a long time since I did this, but the role of the aperture
is the same. There is an optimum aperture size for any given beam
current and either a smaller or larger aperture than that optimum will
increase the beam diameter.

Now for the practical. What actually happens when you turn a knob or
change an aperture on a particular instrument depends on the details of
how that instrument was designed and is being operated. When a SEM
employs a "virtual" aperture (the physical aperture doesn't reside in
the principal plane of the probe-forming lens) then it is possible to
change the *effective* aperture size by manipulating the condenser
lens(es) and thus the effective aperture size can be optimized by the
control software to be optimal for that probe current without changing
the physical aperture. Even if that is not the case, it's usually
impractical or inconvenient to optimally match the physical aperture to
its ideal size (apertures tend to be available in rather coarse diameter
steps). Consequently, for modest changes in beam current, just turning
the "Spot" control (or whatever you call the control that regulates the
condenser current) is the simple practical expedient. But in more
extreme cases, where one wants to drastically increase the beam current
but doesn't want to unduly enlarge the spot, one is well advised to pay
attention to the physics -- the principle works.

Now, back to the original question that was asked. "What would be the
benefit of using a larger objective apperture instead of increasing the
probe current in order to increase the beam current for edx analysis and
x-ray mapping?" There are certainly many situations where choosing a
larger aperture size will permit the higher probe current required for
x-ray analysis to be achieved with minimal degradation of beam diameter.
That is a practical reality in many situations. There may be other
good reasons, and this answer won't apply to all situations. But it is
a mechanism that needs to be considered given the limited information
provided. So far from being a "nuance," depending on the instrument
being referenced, this may in fact be the most pertinent answer to the
question posed.

Fred Schamber
ASPEX Corp.

gary-at-gaugler.com wrote:
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}
} This follows published info. However, the question was
} about aperture size. The other variable of condenser
} squeeze or relaxation is something else to throw into the
} pot. We don't know if his system has a condenser control.
} Probably so.
}
} He was looking for increase in probe current. I figure
} that it is safe to say that everything else being equal,
} increasing the aperture size will increase probe current.
} That is really all he wants to do/know--notwithstanding all
} the other nuances. He just wants to know about a larger
} final aperture size.
}
} gary g.
}
}
}
} At 11:55 AM 11/2/2005, you wrote:
}
}
}
}
} } ----------------------------------------------------------------------------
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From: kenconverse-at-qualityimages.biz
Date: Thu, 3 Nov 2005 14:18:35 -0600
Subject: [Microscopy] SEM - repair service

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

Lori,
I've had a number of requests in the past year or so to service LEO
equipment. The problem is that they don't provide schematics and a third
party can't come in and have any hope of being competent without them.

As far as I know, every other manufacturer provides schematics with their
system.

Perhaps someone from LEO would like to comment.

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: laable-at-solutia.com [mailto:laable-at-solutia.com]
Sent: Wednesday, November 02, 2005 2:34 PM
To: kenconverse-at-qualityimages.biz

Is anyone aware of an independent company who offers yearly service
contracts
on LEO field emission SEMS. I would appreciate any info possible.

Thanks,
Lori Ables
laable-at-solutia.com


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From: ramadanhany-at-gmail.com
Date: Thu, 3 Nov 2005 20:10:34 -0600
Subject: [Microscopy] A favor needed....Thin film technology handbook....

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

Hi guys,

I am writing to see if there is anyone has the following book:
******
Thin film technology handbook
By: Aicha A. R. Elshabini-Riad, Fred D. Barlow III.
c1998
******

It is not available now at the university library and all what I need
is to read pages 27:34, so I hope if there is someone has it and scan
those pages and send them to me. I really need them urgently.


Thanks in advance and I appreciate your time and help.

Best Regards

Hany

--
**********************************************************
Hany Ramadan
Graduate student
Chemistry department
McMaster university, Hamilton, Ontario, Canada
905-525-9140 x: 26322
elsayeh-at-mcmaster.ca
**********************************************************


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9, 23 -- From: Hany Ramadan {ramadanhany-at-gmail.com}
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9, 23 -- Subject: A favor needed....Thin film technology handbook....
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From: akoorts-at-medic.up.ac.za
Date: Fri, 4 Nov 2005 07:37:28 -0600
Subject: [Microscopy] [Filtered] AskAMicroscopist: Grid support films

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This Question was submitted to Ask-A-Microscopist by (akoorts-at-medic.up.ac.za)
from http://www.msa.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, November 4, 2005 at 01:39:39
Remember to consider the Grade/Age of the student when considering the Question
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Email: akoorts-at-medic.up.ac.za
Name: Alida Koorts

Organization: University of Pretoria

Education: Graduate College

Location: South Africa

Title: Grid support films

Question: I loose a lot of sections from uncoated 200 mesh nickel grids during immunolocalization (including autoclave antigen retrieval) and in situ hybridization (including autoclave pre-treatment). Suggestions to appropriate support films that would not interfere with the immunolocalization and in situ hybridization reactions?

Regards

Alida

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From: marta.taules-at-uab.es
Date: Fri, 4 Nov 2005 07:38:46 -0600
Subject: [Microscopy] viaWWW: purity of ethane for cryo fixation?

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Email: marta.taules-at-uab.es
Name: Marta Taules

Organization: UAB

Title-Subject: [Filtered] MListserver:

Question: Dear collegues,

I would like to know which should be the purity of ethane in order to apply the immersion cryo fixation with forceps injector (CPC equipment from Leica).

Thank you in advanced

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From: ramadanhany-at-gmail.com
Date: Fri, 4 Nov 2005 12:28:42 -0600
Subject: [Microscopy] Re: A favor needed....Thin film technology handbook....

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Thanks Jim for your email, here are the details:
This book is available at the library but it is checked out and it may
take more than 10 days to recall it and I need it for a report that is
due in a few days. I can not use ILL to get it because the library
already has it but it is just checked out.

Thanks

Best Regards

Hany


On 11/4/05, Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} wrote:
} Hany
}
} Your library has a ILL (Inter Library Loan) department
} that will do that for you.
}
} regards,
}
} Jim
}
} } From mail-at-ns.microscopy.com Thu Nov 3 21:14:19 2005
} } Return-Path: {mail-at-ns.microscopy.com}
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} } Thu, 3 Nov 2005 20:12:11 -0600
} } Date: Thu, 3 Nov 2005 20:12:11 -0600
} } Message-Id: {200511040212.jA42CBDk029591-at-ns.microscopy.com}
} } To: jquinn-at-www.matscieng.sunysb.edu
} } From: ramadanhany-at-gmail.com
} } Reply-to: ramadanhany-at-gmail.com
} } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com}
} } Subject: [Microscopy] A favor needed....Thin film technology handbook....
} } Errors-To: MicroscopyListSpamFilter-at-microscopy.com
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} } ----------------------------------------------------------------------------
} }
} } Hi guys,
} }
} } I am writing to see if there is anyone has the following book:
} } ******
} } Thin film technology handbook
} } By: Aicha A. R. Elshabini-Riad, Fred D. Barlow III.
} } c1998
} } ******
} }
} } It is not available now at the university library and all what I need
} } is to read pages 27:34, so I hope if there is someone has it and scan
} } those pages and send them to me. I really need them urgently.
} }
} }
} } Thanks in advance and I appreciate your time and help.
} }
} } Best Regards
} }
} } Hany
} }
} } --
} } **********************************************************
} } Hany Ramadan
} } Graduate student
} } Chemistry department
} } McMaster university, Hamilton, Ontario, Canada
} } 905-525-9140 x: 26322
} } elsayeh-at-mcmaster.ca
} } **********************************************************
} }
} }
} } ==============================Original Headers==============================
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}


--
**********************************************************
Hany Ramadan
Graduate student
Chemistry department
McMaster university, Hamilton, Ontario, Canada
905-525-9140 x: 26322
elsayeh-at-mcmaster.ca
**********************************************************


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From: guderjan-at-yahoo.com
Date: Fri, 4 Nov 2005 15:51:47 -0600
Subject: [Microscopy] Zeiss Model 109

Contents Retrieved from Microscopy Listserver Archives
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Maya Research Program is a non-profit corporation that
supports archaeological research and is affiliated
with Texas Christin University. We have been given a
Zeiss Model 109 SEM. It was taken out of service by
the University of Houston, but apparently is in very
good condition.

We wish to sell this instrument. Does anyone know of
someone who deals in such equipment?

Many thanks in advance.

Tom Guderjan





__________________________________
Yahoo! Mail - PC Magazine Editors' Choice 2005
http://mail.yahoo.com

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From: ac.richardson2-at-btinternet.com
Date: Sat, 5 Nov 2005 12:32:35 -0600
Subject: [Microscopy] TEM: freeze substitution

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Hello all,

A quick technical question - we are trying to do freeze substitution using
2%Osmium in acetone, but it goes black as we warm from -90 to -60. Have
you done this and if so is this normal or is there something wrong with
our Os/acetone? Acetone has been kept over a molecular sieve in dialysis
tubing,
I have read somewhere that molecular sieves should be avoided, could
this be the problem?
Acetone is at -20c before adding to osmium and then immediately frozen.





A.C.Richardson
Experimental Officer
School of Biological and Biomedical Science
Centre for Molecular Imaging
University of Durham
Science site
South Rd
Durham
England
E-mail: a.c.richardson-at-durham.ac.uk

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From: dsherman-at-purdue.edu
Date: Sat, 5 Nov 2005 15:54:57 -0600
Subject: [Microscopy] Fixing insect eyes

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We have a project coming up for preparation of mosquito eyes for both LM and
TEM examination. I was planning on using a glut-PAF fix in cacodylate
buffer followed by osmium tetroxide, ETOH dehydration and embedding in epoxy
generic resin.

Options are incubation with tannic acid and en bloc staining with UA prior
to dehydration.

I would appreciate some suggestions as to preparation protocols. Have any
of you tried the tannic acid incubation? If so, at what concentration and
when...in both glut/PAF and Os or alone after glut/PAF? Have you used it at
RT or 4oC? Has en bloc UA been helpful in increasing contrast in eye tissue?

Any suggestions would be most welcome.

Thanks,
Debby

Debby Sherman, Manager 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://www3.agriculture.purdue.edu/microscopy


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From: dswilliams-at-ucsd.edu
Date: Sun, 6 Nov 2005 12:34:18 -0600
Subject: [Microscopy] viaWWW: POSTDOCTORAL POSITION in cell biology @ UCSD

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Email: dswilliams-at-ucsd.edu
Name: DS Williams

Organization: UCSD

Title-Subject: [Filtered] POSTDOCTORAL POSITION in cell biology -at- UCSD

Question: POSTDOCTORAL POSITION in cell biology
UCSD SCHOOL OF MEDICINE
LA JOLLA, CALIFORNIA

Experience in microscopy required, preferably electron microscopy. Studies will be on cellular mechanisms in the retina and potential therapies for retinal degeneration. They will also include collaborative studies on other neurodegenerative disorders. See lab web site for further information about the lab: http://medicine.ucsd.edu/williams/. There will be ample opportunity to develop state-of-the-art skills and collaborations in the rich scientific environment of La Jolla. Start date: negotiable. Salary: NIH scale. Contact: Dr David Williams, Departments of Pharmacology and Neurosciences, UCSD School of Medicine, La Jolla, CA 92093-0912. Email: {DSWILLIAMS-at-UCSD.EDU}


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From: richard.beanland-at-bookham.com
Date: Mon, 7 Nov 2005 05:56:40 -0600
Subject: [Microscopy] RE: Reflecting objectives

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Hi Folks,
while rooting around in a cupboard for an eyepiece with graticule this morning I came across a couple of Beck "reflecting objectives" in their little mahogany boxes. I would dearly like to know what they are used for - it's clear that they focus parallel light to a point... Do they have a use in microscopy, or have I just found a component from a rather old experiment on an optical bench?

Many thanks

Richard

________________________________________
Richard Beanland
Analytical Services
Bookham Inc
Caswell
Towcester
Northants
NN12 8EQ
United Kingdom
Tel. +44 1327 356362
Fax. +44 1327 356775
http://www.bookham.com
________________________________________

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From: frank.karl-at-degussa.com
Date: Mon, 7 Nov 2005 06:44:54 -0600
Subject: [Microscopy] Reflecting objectives

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

Reflective objectives had several different proposes. One was an attempt
to create better images as compared to the crude glass lens of the late
1800 early 1900's. They also had a purpose as they would gather and focus
IR and UV. Of course you needed to use film to collect and "see" the
image. I seem to remember that later ones sometimes contained a glass lens
to assist in correcting for visible light images. Of course you lost all
the UV /I R that the glass filtered.

Modern IR collecting objectives used in micro spectroscopy are reflective
objectives revisited and we come full circle....

Frank Karl
Degussa Corporation
Akron Technical Center
3500 Embassy Parkway
Suite 100
Akron, Ohio 44333


330-668-2235 Ext. 238


This e-mail transmission, and any documents, files or previous e-mail
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richard.beanland-at-
bookham.com To: frank.karl-at-degussa.com
cc:
11/07/2005 06:58 Subject: [Microscopy] RE: Reflecting objectives
AM
Please respond to
richard.beanland








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

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


Hi Folks,
while rooting around in a cupboard for an eyepiece with
graticule this morning I came across a couple of Beck "reflecting
objectives" in their little mahogany boxes. I would dearly like to know
what they are used for - it's clear that they focus parallel light to a
point... Do they have a use in microscopy, or have I just found a component
from a rather old experiment on an optical bench?

Many thanks

Richard

________________________________________
Richard Beanland
Analytical Services
Bookham Inc
Caswell
Towcester
Northants
NN12 8EQ
United Kingdom
Tel. +44 1327 356362
Fax. +44 1327 356775
http://www.bookham.com
________________________________________

=======================================================================
This e-mail is intended for the person it is addressed to only. The
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From: stranen_connelly-at-yahoo.com
Date: Mon, 7 Nov 2005 08:00:11 -0600
Subject: [Microscopy] Re: Fixing insect eyes

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

Debbie,

I have done work with Drosophila eyes and the
following points may be of interest to you:

There are bad results if one fixes the whole head for
not enough fix gets through the "neck".

UA does really increase the contrast. My way was 1%UA
in the refrigerator over night (cold and dark) since
specimens were usually ready to be fixed in the
afternoon and I had used a combination of glut. and
OsO4 on ice for fixation, but an hour or so also
works.

If looking at nerves or cytoskeleton a (good quality)
Acetone dehydration works better than EtOH.

The tannic acid might give some interesting results
but it was not tried in my experiments. If I recall
correctly when using tannic acid it was with the glut.
at room temp. and freshly made but others will no
doubt be better on this point.

Standard epoxy embedding is fine.

I had wanted to try the microwave techniques on the
insect heads to see if the head dissection could be
avoided - everything in its proper place - but did not
have access to a machine until after the head
experiments were over.

Lots of luck!
Pat Connelly
stranen_connelly-at-yahoo.com
(currently seeking employment)

--- dsherman-at-purdue.edu wrote:

} We have a project coming up for preparation of
} mosquito eyes for both LM and
} TEM examination. I was planning on using a glut-PAF
} fix in cacodylate
} buffer followed by osmium tetroxide, ETOH
} dehydration and embedding in epoxy
} generic resin.
}
} Options are incubation with tannic acid and en bloc
} staining with UA prior
} to dehydration.

} I would appreciate some suggestions as to
} preparation protocols. Have any
} of you tried the tannic acid incubation? If so, at
} what concentration and
} when...in both glut/PAF and Os or alone after
} glut/PAF? Have you used it at
} RT or 4oC? Has en bloc UA been helpful in increasing
} contrast in eye tissue?

} Debby Sherman, Manager 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://www3.agriculture.purdue.edu/microscopy





__________________________________
Yahoo! Mail - PC Magazine Editors' Choice 2005
http://mail.yahoo.com

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From: bharris-at-uoguelph.ca
Date: Mon, 7 Nov 2005 10:06:19 -0600
Subject: [Microscopy] networking EM computers

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

Good Morning: We have 2 TEMs with a 3rd coming that are acquiring digital
images. We would like to store these images centrally and deliver them to
customers over the internet. To this end we have purchased the SIS Web Racer
program but don't know how to continue. Our IT dept won't support us. If anyone
out there is doing this I would much appreciate a note on how to construct such
a network from the ground up. Thanks bob

Guelph Regional Imaging Facility
Dept.of Molecular and Cellular
Biology
New Science Complex
488 Gordon St.
Univ of Guelph
Guelph,On, Canada
N1G 2W1
ph: 519-824-4120 ext. 56409/58962
Fax: 519-837-1802

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From: john.mardinly-at-intel.com
Date: Mon, 7 Nov 2005 11:09:15 -0600
Subject: [Microscopy] Reflecting objectives

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

Dear Bob,

Have at look at the Open Microscopy Environment - Jason Smedlow of OME gave
a talk at UCL recently about doing such a thing using "a single relational
database with XML for data migration". See the www.openmicroscopy.org . I
don't know if they can provide one-to-one help but they would seem a good
starting point as they have such a system up and running (and I think
received a fair bit of funding to set it up).

Regards

Keith

----------------------------------------------------------
Dr Keith J Morris
Imaging Facilities Manager
Cell Biology Division
Institute of Ophthalmology
University College London
11-43 Bath Street
London EC1V 9EL

Tel: 020 7608 4050
Fax: 020 7608 4034
email: keith.morris-at-ucl.ac.uk


----- Original Message -----
X-from: {bharris-at-uoguelph.ca}
To: {keith.morris-at-ucl.ac.uk}
Sent: Monday, November 07, 2005 4:14 PM


Another useful aspect of reflective objectives is that they can maintain
a large NA at a long working distance and hence get better resolution
than glass lenses at similar long working distance. Kind of like a
telescope......

John Mardinly
Intel

The copinions of the writer are not necessarily the opinions of Intel
Corporation.

-----Original Message-----
X-from: frank.karl-at-degussa.com [mailto:frank.karl-at-degussa.com]
Sent: Monday, November 07, 2005 4:45 AM
To: Mardinly, John

Reflective objectives had several different proposes. One was an
attempt
to create better images as compared to the crude glass lens of the late
1800 early 1900's. They also had a purpose as they would gather and
focus
IR and UV. Of course you needed to use film to collect and "see" the
image. I seem to remember that later ones sometimes contained a glass
lens
to assist in correcting for visible light images. Of course you lost
all
the UV /I R that the glass filtered.

Modern IR collecting objectives used in micro spectroscopy are
reflective
objectives revisited and we come full circle....

Frank Karl
Degussa Corporation
Akron Technical Center
3500 Embassy Parkway
Suite 100
Akron, Ohio 44333


330-668-2235 Ext. 238


This e-mail transmission, and any documents, files or previous e-mail
messages attached to it may contain information that is confidential or
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 you must not read this transmission and that any
disclosure,
copying, printing, 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
by telephone or return e-mail and delete the original transmission and
its
attachments without reading or saving in any manner. Thank you.




richard.beanland-at-

bookham.com To:
frank.karl-at-degussa.com

cc:

11/07/2005 06:58 Subject: [Microscopy]
RE: Reflecting objectives
AM

Please respond to

richard.beanland










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

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


Hi Folks,
while rooting around in a cupboard for an eyepiece with
graticule this morning I came across a couple of Beck "reflecting
objectives" in their little mahogany boxes. I would dearly like to know
what they are used for - it's clear that they focus parallel light to a
point... Do they have a use in microscopy, or have I just found a
component
from a rather old experiment on an optical bench?

Many thanks

Richard

________________________________________
Richard Beanland
Analytical Services
Bookham Inc
Caswell
Towcester
Northants
NN12 8EQ
United Kingdom
Tel. +44 1327 356362
Fax. +44 1327 356775
http://www.bookham.com
________________________________________

=======================================================================
This e-mail is intended for the person it is addressed to only. The
information contained in it may be confidential and/or protected by
law. If you are not the intended recipient of this message, you must
not make any use of this information, or copy or show it to any
person. Please contact us immediately to tell us that you have
received this e-mail, and return the original to us. Any use,
forwarding, printing or copying of this message is strictly prohibited.
No part of this message can be considered a request for goods or
services.
=======================================================================


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From: gary-at-gaugler.com
Date: Mon, 7 Nov 2005 12:07:21 -0600
Subject: [Microscopy] Re: networking EM computers

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

If your IT department won't help you, what do they actually
do? What purpose do they serve? That aside, you will need
some info from them. Do you already have a CAT-5 10/100BaseT
Ethernet connection to the Internet? If not, you will need one.
Then you will need to know if a computer connected to the cable
needs a static IP or if the main server does DHCP. Then you
need to know the DNS servers (primary and secondary) that your
place uses (you need their IP addresses).

Then, get a simple router like Linksys BEFSR41. This is a four
port switch router. Connect Internet cable to the router and
the other computers to the switch ports. I assume that each
PC has a NIC or built-in Ethernet. Use browser to connect to
router, configure it according to info and set up a private
Intranet either via DHCP or static addresses. Static is usually
preferred since you can define machine names directly in
c:\Windows\system32\drivers\etc\hosts, that way, everyone will
get the same copy of hosts and know what is what.

One machine will always have to be on for this to work. Whichever
this one is, use one of its disks as the distribution disk or
connect a USB/Firewire external disk or a removable disk bay in
the PC box. This will be your server PC.

All of the PCs can talk to each other and send and fetch data
from the "server" PC and send to Internet destinations.

Just an idea/suggestion. But this works.

gary g.




At 08:09 AM 11/7/2005, you wrote:


} Good Morning: We have 2 TEMs with a 3rd coming that are acquiring digital
} images. We would like to store these images centrally and deliver them to
} customers over the internet. To this end we have purchased the SIS Web Racer
} program but don't know how to continue. Our IT dept won't support
} us. If anyone
} out there is doing this I would much appreciate a note on how to
} construct such
} a network from the ground up. Thanks bob
}
} Guelph Regional Imaging Facility
} Dept.of Molecular and Cellular
} Biology
} New Science Complex
} 488 Gordon St.
} Univ of Guelph
} Guelph,On, Canada
} N1G 2W1
} ph: 519-824-4120 ext. 56409/58962
} Fax: 519-837-1802
}
} ==============================Original Headers==============================
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From: gcc-at-couger.com
Date: Mon, 7 Nov 2005 14:03:23 -0600
Subject: [Microscopy] Reflecting objectives

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



richard.beanland-at-bookham.com wrote:

}
} Hi Folks,
} while rooting around in a cupboard for an eyepiece with graticule
;this morning I came across a couple of Beck "reflecting objectives" in
;their little mahogany boxes. I would dearly like to know what they
;are used for - it's clear that they focus parallel light to a point...

;Do they have a use in microscopy, or have I just found a component
;from a rather old experiment on an optical bench?

Hi Richard

The are almost certainly microscope objectives. Put one on a scope and see
how it works. The principle behind them is mirrors have almost the same
focal the same focal length over a very wide range of frequencies. The
small exception is explained on this Canon page on Near Field Microscopy
http://www.canon.com/technology/s_labo/light/004/01.html In the Dripping
Light selection it shows the a reflected wave is leaves the surface 1 wave
length away from where it strikes the surface. But for most practical uses
that does not effect us.

If they are low power and low NA the best way to use them for near infra
red and the UV that silicone sensors will capture is to see if they will
work as a prime lens for a video camera with no lenses to have different
focal points at 1000 nm light and 300nm light. The lens will cover a much
wider range of light than silicone will detect but you start taking about
a great deal of money for cameras. You also need a monochromator or filers
to make much sense of the images.

If you need higher resolution than a video camera will record. Three are
some modifiable 3 mega pixel camera such as the Aiptek 5100 that has
smile lens that is easy to replace that will 11 frames per second or 640 x
480 Video and a true 3 Mega pixel image on a memory card for $100. I have
on my desk I am converting. I am not sure if the IR/UV filter is in the
lens or on the sensor. But most of the fixed lens 3 mega pixel cameras can
be modified to take other lenses and the IR/UV filters removed.

If the Breck objectives are below 40x and you are ever going to put up for
surplus I want to be first on the list to be notified when you get rid of
them.

Gordon
Gordon Couger

I collect links on information related to light microscopes.
www.couger.com/microscope/links/gclinks.html
Please forward anything you think might be useful to others.
Microscope Documentation is at www.science-info.org


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From: sghoshro-at-NMSU.Edu
Date: Mon, 7 Nov 2005 14:18:12 -0600
Subject: [Microscopy] Hepatitis C infected tissue

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

Hello Everyone,

A graduate student is planning to fix hepatitis C infected human liver
tissue for TEM (fixation and embedding) work. I would like to know what
would be the best way to fix the fresh tissue and transport it from a
hospital. I am also interested to find out what precautionary measure we
should take since the student will be handling infected liver tissue. We
are planning to fix in glutaraldehyde in cacodylate/phosphate buffer. Is
the virus considered infectious after fixation? Any suggestion will be
greatly appreciated.

Thanks in advance.

Soumitra




*************************************************************
Soumitra Ghoshroy
College Associate Professor, Biology
Director, Electron Microscopy Lab
Box 3EML
New Mexico State University
Las Cruces, NM 88003
Tel: 505-646-3268 (office), 646-3283 (lab)
Fax: 505-646-3282
e-mail:sghoshro-at-nmsu.edu
http://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm
http://emldata.nmsu.edu/eml/

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From: lkrupp-at-us.ibm.com
Date: Mon, 7 Nov 2005 16:05:44 -0600
Subject: [Microscopy] Where to buy epo-tek epoxy

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


Hello-

I need to purchase Epo-tek 350ND epoxy, and am having trouble finding it
online. Does anyone know someone that carries it, or another contact
person?

Vendor replies are fine.

Thank you,
Leslie

Leslie Krupp (Thompson)
IBM Almaden Research
650 Harry Road, K19/D2
San Jose, CA 95120-6099


==============================Original Headers==============================
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From: lkrupp-at-us.ibm.com
Date: Mon, 7 Nov 2005 16:48:21 -0600
Subject: [Microscopy] Fw: Where to buy epo-tek epoxy

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


Leslie Krupp (Thompson)
IBM Almaden Research
650 Harry Road, K19/D2
San Jose, CA 95120-6099
(408) 927-3856
----- Forwarded by Leslie E Krupp/Almaden/IBM on 11/07/2005 02:46 PM -----
|---------+----------------------------}
| | Dmrelion-at-aol.com |
| | |
| | 11/07/2005 02:45 |
| | PM |
|---------+----------------------------}
} -------------------------------------------------------------------------------------------------------------------------------|
| |
| To: Leslie E Krupp/Almaden/IBM-at-IBMUS |
| cc: |
| Subject: Re: [Microscopy] Where to buy epo-tek epoxy |
} -------------------------------------------------------------------------------------------------------------------------------|




Leslie,

Try going to

http://www.epotek.com/categories.asp?ID=3


The list a 353ND but their sales dept. should be able to help.

We buy their Epotek301 for use in mounting geological thin sections for
cathodoluminescence studies. Good optical and thermal properties and stands
up to the heat of the electron beam.

Don Marshall

Donald J. Marshall (Dr.)
RELION Industries
PO Box 12
Bedford, MA 01730
USA

781-275-4695 (phone)
781-271-0252 (FAX)

dmrelion-at-aol.com

http://www.excitingelectrons.com

"A weed is a flower out of place

In a message dated 11/7/2005 5:08:51 PM Eastern Standard Time,
lkrupp-at-us.ibm.com writes:

Hello-

I need to purchase Epo-tek 350ND epoxy, and am having trouble finding it
online. Does anyone know someone that carries it, or another contact
person?

Vendor replies are fine.

Thank you,
Leslie

Leslie Krupp (Thompson)
IBM Almaden Research
650 Harry Road, K19/D2
San Jose, CA 95120-6099

."


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From: gsosinsky-at-ucsd.edu
Date: Tue, 8 Nov 2005 08:09:47 -0600
Subject: [Microscopy] viaWWW: Postdoctoral/Staff position at the National Center for

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Email: gsosinsky-at-ucsd.edu
Name: Gina Sosinsky

Organization: University of California, San Diego

Title-Subject: [Filtered] MListserver: Position available

Question: Postdoctoral/Staff position at the National Center for Microscopy and Imaging Research at the University of California, San Diego, CA

A postdoctoral or staff position in cryo-EM and 3D reconstruction to study connexin26 wild type and mutant structures is available immediately.
Our laboratory has isolated connexin26 gap junctions as in situ ordered two-dimensional crystals and as isolated hemichannels for analysis by electron microscopic structure determination, atomic force microscope imaging and biochemical studies. We are investigating the structure of wild type connexin26 connexons, functional mutants and treatments that result in changes in the pore size or shape to study structural mechanisms of connexon gating.

The successful candidate will have been awarded a Ph.D. or submitted (with oral examination pending), have experience in electron cryo-microscopy and image processing or three-dimensional reconstruction techniques. Experience in membrane purification techniques is preferred, but not essential. Excellent written and verbal skills are expected. The candidate will be working in a large interdisciplinary research center located in the University of California San Diego main campus and will have access to state of the art electron microscopes. For more information on our laboratory, please visit http://www.ncmir.ucsd.edu.
Please forward this message to all appropriate personnel. Applicants should send curriculum vitae, bibliography, a brief description of present research activities and plans and the names and contact information of three references to:

Dr. Gina Sosinsky

University of California at San Diego
1070 Basic Science Building MC 0608
9500 Gilman Drive
La Jolla, CA 92093-0608
858-534-0128 (office phone & voice mail)
858-534-4583 (lab phone)
858-534-7497 (fax)
gsosinsky-at-ucsd.edu (email)


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From: m_jarnik-at-fccc.edu
Date: Tue, 8 Nov 2005 08:10:51 -0600
Subject: [Microscopy] viaWWW: Embedding melanocytes

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Email: m_jarnik-at-fccc.edu
Name: Michal Jarnik

Organization: FCCC

Title-Subject: [Filtered] Embedding melanocytes

Question: We have some problems embedding melanocytes in Epon. Although the overall ultrastructure of the cells is typically good, the melanosomes lack the outer membrane. We tried several varieties of our standard embedding protocol (glutaraldehyde fixation followed by osmication, dehydration in EtOH and propylenoxide and embedding), but no real difference. Any suggestions would be welcome. Thanks.

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From: TindallR-at-missouri.edu
Date: Tue, 8 Nov 2005 10:13:10 -0600
Subject: [Microscopy] TEM: Intractable Pepper Redux

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Hi Soumitra,

Your question was outside my experience, so I contacted a
virologist/microscopist I know, and she kindly sent me the following.

The required precautions sound quite serious and she suggested you
contact her with any questions.

Mike
------------
Any clinical specimen should be handled with "Universal Precautions".

There is a set of guidelines known to anyone who routinely collects
potentially
infectious material. I would let this person place the tissue into
fixative. These procedures entail the use of gloves, lab coat, face
protection, special procedures for not cutting or sticking oneself with
sharp objects, and use of a biosafety level 2 cabinet.

Whoever obtains the specimen from the patient (the surgeon? a pathology
technologist?) can place it directly into glutaraldehyde for transport.
This
will render the specimen non-infectious since the agent is an enveloped
virus
(flavivirus). (Prion material is not killed by aldehyde fixation, but all
human viruses are.) Ask the person obtaining the sample to cut it into
small
pieces or slivers if possible (a few mm). If you get a large chunk (e.g., a
cubic cm), let it sit in glutaraldehyde overnight to ensure that the
fixative
gets inside. Then take your 1 cubic mm slices for EM from the surface
that is
well fixed. The center may or may not be well fixed, depending on how
long the tissue sat before being placed into the glut. At this stage,
you can handle the
tissue just like any other non-infectious specimen.

Note that few good micrographs of HCV have been published, particularly from
infected human liver. Also, note that flaviviruses are small (40-60 nm)
enveloped RNA viruses, and that other things in the cell can resemble these
particles. Most diagnoses of HCV are made histologically on the
appearance of
the liver, by immunostaining, and/or serologically, but not by EM. Good
luck.


Sara E. Miller, Ph. D.
P. O. Box 3712
Duke University Medical Center
Durham, NC 27710
Ph: 919 684-3452
FAX: 919 684-3265


----- Original Message -----
X-from: sghoshro-at-NMSU.Edu

Dear Listers,

Back in the mists of history, about two or three years ago, I put a plea
for help on the listserver concerning pepper-like section contamination.
This stuff resisted every attempt to get rid of it---different
fixatives, four different buffer systems, various resins, five
different water supplies, stains (or no stains!), fixation with and
without osmium, different chemical suppliers, rewashing every piece of
glassware in the lab, prayer, curses, voodoo. Even a Kitchen Witch. We
tried all of the many suggestions from the dedicated denizens of the
list (which normally cure any problem we have), all to no avail. It
showed up in some tissues all of the time (especially retina), some
tissues some of the time, some tissues never, and was often localized on
membranes. We finally determined that it was osmium in part (although
we got some pepper even in unosmicated samples). So I discussed the
problem with a chemist who specialized in osmium chemistry. He mumbled
something about changes in the air in the lab and wished us luck as he
beat a hasty retreat. Must have been the haunted look on my face.

Finally, after almost two years, in desperation our lab director
suggested adding 0.01M 2-mercaptoethanol to the buffer washes after the
primary fixation step and in the osmium step. No more pepper. I
promised if we ever figured this one out, I would share it with the
list, so here you go.

The problem is that we never really figured it out, except for how to
make it go away. That's great on one hand, but fundamentally
frustrating on the other, since we never got a handle on why the
contamination appeared suddenly in the first place and took up residence
in the lab.

So, if you too have Intractable Pepper Syndrome, try the 2-ME. If you
have any thoughts on why this works, we (and Tom Phillips) would love to
hear them.

Relieved, but puzzled,
Randy

Randy Tindall
EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W122 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







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From: phillipst-at-missouri.edu
Date: Tue, 8 Nov 2005 10:22:30 -0600
Subject: [Microscopy] Pepper artefact

Contents Retrieved from Microscopy Listserver Archives
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I have developed a major problem with the infamous pepper artefact. I have
small intensely black dots associated with the microvilli, rough ER, Golgi,
nuclear envelope, and plasma membrane. The spots vary some in size from
about 6 to 30 nm. They are not on the nucleus or mucin granules except
along their peripheral membranes. Tissues were incubated in vivo with a
biotinylated antibody which would label an apical membrane surface antigen
and then removed and rinsed in 6x in PBS. Fix in 2% PF + 0.2%
glutaraldehyde + 300 nM DAPI in HWB (30 mM HEPES, 70 mM NaCl, 2 mM CaCl2,
pH 7.4) for overnight at 4 C. I need the DAPI to identify the region of
interest prior to dissection. Rinse in HWB + glycine 3 x 10 min. Dissect
out region of interest. Block in 0.1% acetylated BSA for 45 min. Incubate
the samples in a 1:10 dilution of 10 nm gold conjugated goat anti-biotin in
0.1% BSA-c in HWB for overnight at 4 C. Rinse 6x 5 min in HWB. 30 min in
25% ethanol + 0.5% uranyl acetate. 50% , 70%, 95% at 4 C, 3 x 100% ethanol
for 10 min each at -20 °C. 1:1 LR Gold (no catalyst):ethanol 8 hrs at 4
°C. 2:1 LRG (no catalyst):ethanol overnight at 4 C. 100% LRG + catalyst
overnight at -20 °C. 100% LRG + catalyst overnight at -20 °C. 100% LRG +
catalyst for 2 hrs at -20 °C. Embed in 200 ul of fresh LRG +
catalyst in sealed BEEM capsules at 4 C under UV light. Collect ultrathin
sections (50–80 nm) on nickel grids. The problem is present regardless of
whether I use lead or uranyl acetate to stain the thin sections.

I can't remove the pepper by floating the grids on either 0.5% HCl, 1% EDTA
or 2% periodic acid for up to 60 min. I am guessing it is from the uranyl
acetate en bloc but the artefact is small round dots which is not what
uranyl artefacts have looked like in earlier failures. I am confident is
not a gold artefact since gold would not penetrate into the cells and I can
recognize a colloidal gold particle. Any thoughts on where this is coming
from or how to remove it from my existing blocks would be welcome.


Thomas E. Phillips, PhD
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




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From: phillipst-at-missouri.edu
Date: Tue, 8 Nov 2005 11:40:38 -0600
Subject: [Microscopy] Quetol

Contents Retrieved from Microscopy Listserver Archives
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I am going to bite the bullet and try another embedding resin for a number
of reasons. I want to give Quetol a try because of its reported nice
contrast and ability to tolerate a small amount of water. The
manufacturers data sheet gives a formulation using volume measurements
(mls) of each component. I must prefer to use the more accurate and
convenient method of weighing the components. Does anyone have a good
Quetol formulation by weight? I googled it and found John Johnson uses:
15 gm Quetol 651
25 gm NSA
4 gm MNA
1 gm DMP-30
but others seem to have quite different proportions. Any comments?



Thomas E. Phillips, PhD
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



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From: twigg-at-estd.nrl.navy.mil
Date: Tue, 8 Nov 2005 11:50:02 -0600
Subject: [Microscopy] Grinding Tinanium Carbide for XTEM

Contents Retrieved from Microscopy Listserver Archives
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We are trying to prepare XTEM samples from tinanium carbide
substates, but the 180 grit SiC paper we use in the initial grinding
step is just not up to taking on this extremely hard material. Any
suggestions (including vendors for such abrasives)?
--
Dr. Mark E. Twigg
Code 6812
Naval Research Laboratory
4555 Overlook Ave., S.W.
Washington DC 20375

tel: (202) 404-8543
fax: (202) 404-7194

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From: randerson20-at-tampabay.rr.com
Date: Tue, 8 Nov 2005 12:16:27 -0600
Subject: [Microscopy] Re: Grinding Tinanium Carbide for XTEM

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

twigg-at-estd.nrl.navy.mil wrote:

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

Oh, and patience. Lots of patience.

Ron Anderson


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From: hyi-at-emory.edu
Date: Tue, 8 Nov 2005 12:22:28 -0600
Subject: [Microscopy] Pepper artefact

Contents Retrieved from Microscopy Listserver Archives
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Hi, Thomas:

If it is not too much trouble, would you send me an image of this
pepper artifact off-line? Thank you in advance.

Hong Yi
Emory EM


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From: edelmare-at-muohio.edu
Date: Tue, 8 Nov 2005 13:16:36 -0600
Subject: [Microscopy] Re: Quetol

Contents Retrieved from Microscopy Listserver Archives
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Thomas:

Here's my formula for Quetol HARD. I've used it numerous times as a
replacment for Spurr's Hard (Quetol version is slightly harder
actually).

Quetol 651 17 g

NMA
(nadic methyl anhydride) 23 g

DMAE / DMP-30
( 2-dimethylaminoethanol or S-1) 0.5 g

=======================================

I am going to bite the bullet and try another embedding resin for a
number
of reasons. I want to give Quetol a try because of its reported nice
contrast and ability to tolerate a small amount of water. The
manufacturers data sheet gives a formulation using volume measurements
(mls) of each component. I must prefer to use the more accurate and
convenient method of weighing the components. Does anyone have a good
Quetol formulation by weight? I googled it and found John Johnson uses:
15 gm Quetol 651
25 gm NSA
4 gm MNA
1 gm DMP-30
but others seem to have quite different proportions. Any comments?



Thomas E. Phillips, PhD
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



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Richard E. Edelmann
Electron Microscopy Facility Director
362 Pearson Hall
Miami University
Oxford, OH 45056
Ph: 513-529-5712
Fax: 513-529-4243
HTTP://www.emf.muohio.edu

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From: smalinskas-at-yahoo.com
Date: Tue, 8 Nov 2005 14:29:57 -0600
Subject: [Microscopy] Grinding Tinanium Carbide for XTEM

Contents Retrieved from Microscopy Listserver Archives
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Like Ron said, diamond abrasive is the way to go.

Another popular source is Struers (www.struers.com)
and Lapmaster (www.lapmaster.com). They both have
diamond discs (rough, 10-250 micron) and diamond films
(fine, 1/4-30 micron).

Stu Smalinskas, P.E.
Metallurgist
SKF USA
Plymouth, Michigan

--- randerson20-at-tampabay.rr.com wrote:

} Diamond lapping film on mylar substrates. Available
} form a number of
} vendors. We were partial to the films made by DuPont
} because of their
} high density of diamond particles, good particle
} size control, and
} adhesion of the diamonds to the substrate. This
} experience is about five
} years old. I'm sure that by now any number of
} manufacturers can provide
} films with similar properties.
}
} Oh, and patience. Lots of patience.
}
} Ron Anderson
}
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
} twigg-at-estd.nrl.navy.mil wrote:
}
}
} } We are trying to prepare XTEM samples from tinanium
} carbide
} } substates, but the 180 grit SiC paper we use in the
} initial grinding
} } step is just not up to taking on this extremely
} hard material. Any
} } suggestions (including vendors for such abrasives)?




__________________________________
Yahoo! FareChase: Search multiple travel sites in one click.
http://farechase.yahoo.com

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From: GBurgess-at-exchange.hsc.mb.ca
Date: Tue, 8 Nov 2005 14:40:40 -0600
Subject: [Microscopy] Printing Digital Micrographs

Contents Retrieved from Microscopy Listserver Archives
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Our clinical diagnostic EM laboratory is considering purchasing a digital
camera for our electron microscope to eliminate a lot of the wet lab photo
finishing expenses. I was wondering in the hospital environment how many
people out there end up printing their digital images for records purposes,
rather than just storing the digital images. To me, it seems like a
redundant and expense step if one already has the digital image, since that
image could in fact be printed at any time.

Garry Burgess
Charge Technologist
Health Sciences Centre
Winnipeg, Canada


This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.

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From: bozzola-at-siu.edu
Date: Tue, 8 Nov 2005 15:50:55 -0600
Subject: [Microscopy] freeze fracture contractual work needed

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A colleague in Chemistry would like to have some freeze-fracture work
done on some liposome samples and was wondering if someone would be
available to do this work and the cost per specimen.

Thank you.

JB
--
##############################################################
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
750 Communications Drive - MC 4402
Southern Illinois University
Carbondale, IL 62901 U.S.A.
Phone: 618-453-3730
Email: bozzola-at-siu.edu
Web: http://www.siu.edu/~image/
##############################################################

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From: TindallR-at-missouri.edu
Date: Tue, 8 Nov 2005 16:37:33 -0600
Subject: [Microscopy] Microtomy: Sorvall knifemaker?

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From: TindallR-at-missouri.edu
Date: Tue, 8 Nov 2005 16:53:37 -0600
Subject: [Microscopy] Microtomy again...

Contents Retrieved from Microscopy Listserver Archives
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Sorry, I don't know what happened with the last message, but my request
was whether anyone had or knew of a surplus Sovall glass knifemaker,
circa 1979, for sale. Or does anyone still service these old units?
We need to replace or repair this machine, which takes 3/8" thick
microtomy glass. Alternatively, are there other available knifemakers
out there that will still use the thick glass?

Thanks. This question should be easier to answer, now that it's
actually in the message.

Cheers,
Randy

Randy Tindall
EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W122 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







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From: Stacey.Andringa-at-uc.edu
Date: Tue, 8 Nov 2005 19:31:47 -0600
Subject: [Microscopy] viaWWW: fading sections

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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: Stacey.Andringa-at-uc.edu
Name: Stacey Andringa

Organization: University of Cincinnati

Title-Subject: [Filtered] MListserver:

Question: We have been using a 1% toulidine blue O in 1% sodium borate as a general stain for all of our thick sections (1-2 microns). We use Cytoseal 60 as the mounting medium. We have noticed that the sections are fading more quickly than they have in the past and sometimes we get "round clear droplets" on the sections.
Can anyone recommend a stain that won't fade so quickly?
Does anyone have a clue as to the nature of the
"droplets"?
Thanks.

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

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From: henriks-at-southbaytech.com
Date: Wed, 9 Nov 2005 00:31:38 -0600
Subject: [Microscopy] Re: Grinding Tinanium Carbide for XTEM

Contents Retrieved from Microscopy Listserver Archives
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Mark:

Ron is 100% correct in saying that the diamond lapping film would be suitable for this application. Having supplied this material to Ron in the past, I can say with some confidence that he is referring to a product manufactured by 3M rather than by Dupont. I would be pleased to provide you with product details as well as some guidance on your specific application. Please feel free to contact me offline.

Best regards-

David

David Henriks
South Bay Techology, Inc.
1120 Via Callejon
San Clemente, CA 92673 USA

TEL =1-949-492-2600
FAX: =1-949-92-1499

email: henriks-at-southbaytech.com


--------------------------------------------------------------------------------
X-from: randerson20-at-tampabay.rr.com
Sent: Tuesday, November 08, 2005 1:19 PM
To: henriks-at-southbaytech.com

twigg-at-estd.nrl.navy.mil wrote:

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

Oh, and patience. Lots of patience.

Ron Anderson

David Henriks
South Bay Techology, Inc.
1120 Via Callejon
San Clemente, CA 92673 USA

TEL =1-949-492-2600
FAX: =1-949-92-1499

email: henriks-at-southbaytech.com




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From: W.Muss-at-salk.at
Date: Wed, 9 Nov 2005 03:51:23 -0600
Subject: [Microscopy] Re: fading sections

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Good morning all,
dear Mrs Andringa,

the problem with } fading { stains (usually highly alkalinesolutions basic
dyes like Toluidine Blue in borax solution, Azur II-Methylenblue-basic
Fuchsin according to Richardson and others) on plastic (resin) semithin
sections for me is a long known fact (regardless of the mounting media I
used earlier).....so I started some 20 years ago to change my processing
and archiving mode for semithin sections (we have to store the semithins of
our cases for legal reasons -if possible - up to 20 years).

i) for looking at stained sections for orientation purposes, most of them
also for diagnosing, as well as trimming the right location for
EM-ultrathin sections we do not mount them with a slide and/or mounting
medium.
We find the spatial resolution already achieved WITHOUT mounting a
coverslip (compared to a thick histological, paraffin-embedded section) is
enough to choose the location of ultrathin section trimming.

ii) for a necessary photographic documentation or high resolution light
microscopy of } seclected { stained sections we immerse the respective
sections with a small drop of immersion oil, view either with a coverslide
0.17 mm placed carefully on the oildrop then (e.g. for Obj. x 4, x 10, x
25) or directly with a x 50 or x 100 oil immersion objective (Plan, NA=
1,00; 1,30, respectively....those are the good ones from the 60ies and
70ies....).

After having done this, the coverslide will be slipped away, the object
slide will be immersed in (a) coplin jar(s) filled with xylol (2 x 30 sec.
each) or any other diluent which will solve the immersion oil, afterwards
we give the object slides a blast of compressed air (by means of a nozzle
connected by a tube to a } bottle { or the pipeline of compressed air) and
the mounted sections are ready for storing/archiving.
If you do that in a light protected, dustfree place, you will not see - at
least over 5 years from my experience - a fading of staining, also you will
not have "bad wrinkles" in your sections which are produced over time due
to the mounting agent.

This creates the second advantage, the first being economizing the mounting
process (in avoiding permanent mounting with not very healthy mounting
media as well as saving costs for coverslips and the time needed for
coverslipping, drying of mounts, etc.)
IF (in case this happens) the staining of sections has been gone or at
least does not display brilliantly after a long period of time as you are
expecting, you are able to stain again your sections quite easy and
rapidly with the routine staining procedure without loss of substantial
information.

Unfortunately I am really no chemist and therefore I assume a chemical
reaction (de-composing solvent/material due to } aging {) which produces the
"round clear droplets" you see in your mounted sections.

Best regards and wishes for a solution of your problem,

Wolfgang Muss
Salzburg, Austria

PS: if you would like to have some examples of images taken digitally ith
the method(s) given above, I can send them if you like (please send
allowance message for sending them).

----------
Von: Stacey.Andringa-at-uc.edu[SMTP:Stacey.Andringa-at-uc.edu]
Antwort an: Stacey.Andringa-at-uc.edu
Gesendet: Mittwoch, 09. November 2005 02:36
An: W.Muss-at-salk.at
Betreff: [Microscopy] fading sections (Tolblue in Borax)




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Email: Stacey.Andringa-at-uc.edu
Name: Stacey Andringa

Organization: University of Cincinnati

Title-Subject: [Filtered] MListserver:

Question: We have been using a 1% toulidine blue O in 1% sodium borate as a
general stain for all of our thick sections (1-2 microns). We use Cytoseal
60 as the mounting medium. We have noticed that the sections are fading
more quickly than they have in the past and sometimes we get "round clear
droplets" on the sections.
Can anyone recommend a stain that won't fade so quickly?
Does anyone have a clue as to the nature of the
"droplets"?
Thanks.

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24, 28 -- From W.Muss-at-salk.at Wed Nov 9 03:51:23 2005
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From: Edward.Calomeni-at-osumc.edu
Date: Wed, 9 Nov 2005 07:42:02 -0600
Subject: [Microscopy] Printing Digital Micrographs

Contents Retrieved from Microscopy Listserver Archives
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Garry,

I very rarely print digital images. There are two pathologists who want
prints, so I just give them low resolution, plain paper prints (nothing high
quality) for their own records. All images are stored in a departmental
image database (Paxit). Anyone in our department has access to them.
If images are to be sent out for consultation purposes, I send a CD of the
tif files. In the past 2.5 years, I have printed less than 200 high quality
photo paper digital images.


Ed






Edward P. Calomeni
Director EM Lab
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: GBurgess-at-exchange.hsc.mb.ca [mailto:GBurgess-at-exchange.hsc.mb.ca]
Sent: Tuesday, November 08, 2005 3:46 PM
To: Edward Calomeni


Our clinical diagnostic EM laboratory is considering purchasing a digital
camera for our electron microscope to eliminate a lot of the wet lab photo
finishing expenses. I was wondering in the hospital environment how many
people out there end up printing their digital images for records purposes,
rather than just storing the digital images. To me, it seems like a
redundant and expense step if one already has the digital image, since that
image could in fact be printed at any time.

Garry Burgess
Charge Technologist
Health Sciences Centre
Winnipeg, Canada


This e-mail and/or any documents in this transmission is intended for the
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From: Mike.Bode-at-soft-imaging.net
Date: Wed, 9 Nov 2005 09:41:26 -0600
Subject: [Microscopy] Printing Digital Micrographs

Contents Retrieved from Microscopy Listserver Archives
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Hello Garry,

We work a lot with Pathology departments in Hospitals, and our experience is that most departments are at first reluctant to the idea of not printing anymore. Prints is what the users are used to and what they know how to interpret. In fact, we often find that the Hospitals have very specific instructions about the size and paper that is to be used for printing. However, once the advantages of digital images hit the users, the use of printed images goes down. Especially, if you can equip all stations with a viewer software and have the ability to archive and transmit images to other facilities, it becomes obvious pretty quickly that prints just take a lot of time and are expensive to ship around. And it is pretty cheap to buy a couple of high quality ink jet printers for the occasional print.

However, in order to get there, you need to plan a bit ahead. Use a software that has an archiving or image database and that allows multiple users to use the archive. Make sure that you have the option to allow external users to access the database. Talk to your IT department. Keep future expansions in mind. Check to see if there are any restrictions (for example, in the US there are HIPAA laws, and the FDA may get involved with something called 21 CFR rule 11).

If you do your homework first, you can set up a system that increases your throughput quite a bit. Let me know if we can help you further with any of this.

Mike


Michael Bode, Ph.D.
Soft Imaging System Corp.
12596 West Bayaud Avenue
Suite 300
Lakewood, CO 80228
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-----Original Message-----
X-from: GBurgess-at-exchange.hsc.mb.ca [mailto:GBurgess-at-exchange.hsc.mb.ca]
Sent: Tuesday, November 08, 2005 1:43 PM
To: Mike Bode


Our clinical diagnostic EM laboratory is considering purchasing a digital camera for our electron microscope to eliminate a lot of the wet lab photo finishing expenses. I was wondering in the hospital environment how many people out there end up printing their digital images for records purposes, rather than just storing the digital images. To me, it seems like a redundant and expense step if one already has the digital image, since that image could in fact be printed at any time.

Garry Burgess
Charge Technologist
Health Sciences Centre
Winnipeg, Canada


This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.

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From: glenmac-at-u.washington.edu
Date: Wed, 9 Nov 2005 11:19:37 -0600
Subject: [Microscopy] Re: viaWWW: fading sections

Contents Retrieved from Microscopy Listserver Archives
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Dear Stacey,
The droplets are likely water from incomplete dehydration and
clearing. Cytoseal is a toluene based mounting medium that will not
mix with water. Extend your dehydration times and/or add an
additional absolute ethanol step, and maybe add another step in
clearing agent. Make sure you replace the dehydration baths and
clearing baths on a regular basis as water will gradually build up in
them. Although a greater issue with thicker paraffin or vibratome
sections, it can occur with plastic sections if they are rushed
through or the baths have not been changed.

What is the time frame of your fading? Days, months or years? Çould
it be the pH of the Cytoseal? We get a few years with Toluidine Blue
mounted in DPX.

Regards,
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-u.washington.edu

************************************************************************
******
The box said "Requires Windows 95 or better", so I bought a Macintosh.
************************************************************************
******


On Nov 8, 2005, at 5:33 PM, Stacey.Andringa-at-uc.edu wrote:

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} Question: We have been using a 1% toulidine blue O in 1% sodium
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} microns). We use Cytoseal 60 as the mounting medium. We have
} noticed that the sections are fading more quickly than they have in
} the past and sometimes we get "round clear droplets" on the sections.
} Can anyone recommend a stain that won't fade so quickly?
} Does anyone have a clue as to the nature of the
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} Thanks.
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From: diaspro-at-fisica.unige.it
Date: Thu, 10 Nov 2005 07:51:27 -0600
Subject: [Microscopy] viaWWW: CONFOCAL 7 Meeting

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Email: diaspro-at-fisica.unige.it
Name: Alberto Diaspro

Organization: University of Genoa

Title-Subject: [Filtered] CONFOCAL 7 Meeting

Question: ELAMBS-MICROSCOBIO, DEPARTMENT OF PHYSICS, UNIVERSITY OF GENOA PROUDLY ANNOUNCES THE 7TH PRACTICAL INTENSIVE WORKSHOP ON 3D CONFOCAL MICROSCOPY ENTITLED 3D EXPLORATION OF THE LIVING WORLD

CONFOCAL 7, will be held at LAMBS, GENOA JAN 31-FEb 3, 2006.

KEY SCIENTISTS IN THE FIELD WILL GIVE TALKS ON HOT SUBJECTS IN 3D CONFOCAL MICROSCOPY INCLUDING MULTIPHOTON IMAGING, ìFî TECHNIQUES, BIOMEDICAL APPLICATIONS.

TRAINING WILL BE ON CONFOCAL AND MULTIPHOTON SPECTRAL MICROSCOPES, ULTRAFAST LASER TI-SAPPHIRE LASER SOURCES AND IMAGE DECONVOLUTION WORKSTATIONS. 4D (X-Y-Z-t), FRET, FRAP, FLIM and SHG EXPERIMENTS WILL BE PERFORMED. STUDENTS ARE ENCOURAGED TO BRING THEIR OWN SAMPLES SINCE CELL CULTURE, CHEMICAL, CELL BIOLOGY FACILITY WILL BE AVAILABLE.

Participation fees:

Academia/University Researchers 450 Ä: full School including CD and 2 lunches (Fluorescence basics on demand).

Industry/Private Researchers 600 Ä: full School including CD and 2 lunches

(Fluorescence basics on demand).

Voluntary contribution 50 Ä: Tuesday afternoon Lectures.

FREE: Tuesday afternoon Lectures, for Students, any level, and Junior Researchers.

Practical sessions will be restricted to 20 attendants.

Application: Interested candidates should send an e-mail to the Director of the Course at the following e-mail address: {mailto:diaspro-at-fisica.unige.it} diaspro-at-fisica.unige.it, using the following subject: CONFOCAL 7.

Selection will be realized on a Temporal (first-in-first-out) and Geographical (long distance applicants are favoured) basis. A short Curriculum Vitae and scientific interest of the candidate can be useful for Candidate selection.


For details see EVENTS on www.lambs.it.


---------------------------------------------
[...] Dance with wolves and count the stars, including the
unseen...(L.Ferlinghetti, Challenges to young poet, 2001)
---------------------------------------------
Alberto Diaspro, MicroScoBIO Research Center, LAMBS-IFOM, Department of
Physics, University of Genoa, Via Dodecaneso 33, 16146 Genova, Italy
facsimile +39-010314218 - voice +39-0103536426/480/309;
URL: {http://www.lambs.it} http://www.lambs.it
{http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html} http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html
----------------------------------------------

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From: Anjeanette.Ormonde-at-unilever.com
Date: Thu, 10 Nov 2005 09:42:49 -0600
Subject: [Microscopy] tweezers search

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

I was hoping someone out there might be able to help me out. A co-worker (not
a microscopist) brought me a pair of tweezers and wants to order more but can't
remember where they were originally purchased from. She thinks she bought them
from a microscopy catalogue but we haven't been able to find the same ones in
any of the catalogues I typically order from. They are black plastic (she
think they are carbon-fiber) and the primary concern is that they are
anti-static. While we can find plenty of anti-static, plastic tweezers hers
are curved and our searches keep turning up straight or angled pairs. On one
side is the code "J201" and the flip side has a sort of logo. It looks like a
J surrounded by four "pine trees". Any assistance would be greatly appreciated.

Thanks,
Angie




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5, 14 -- Date: Thu, 10 Nov 2005 10:39:55 -0500
5, 14 -- From: "Anjeanette Ormonde" {Anjeanette.Ormonde-at-unilever.com}
5, 14 -- Subject: tweezers search
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From: W.Muss-at-salk.at
Date: Thu, 10 Nov 2005 10:04:22 -0600
Subject: [Microscopy] Re: tweezers search

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

Dear Angie,

try


http://www.ure.com.my/products/tweezer.html

and find:

J201, curved tips

Hope this helps,

best regards,


Wolfgang Muss
Salzburg, Austria

----------
Von: Anjeanette.Ormonde-at-unilever.com[SMTP:Anjeanette.Ormonde-at-unilever.com]
Antwort an: Anjeanette.Ormonde-at-unilever.com
Gesendet: Donnerstag, 10. November 2005 16:46
An: W.Muss-at-salk.at
Betreff: [Microscopy] tweezers search

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

I was hoping someone out there might be able to help me out. A co-worker
(not
a microscopist) brought me a pair of tweezers and wants to order more but
can't
remember where they were originally purchased from. She thinks she bought
them
from a microscopy catalogue but we haven't been able to find the same ones
in
any of the catalogues I typically order from. They are black plastic (she
think they are carbon-fiber) and the primary concern is that they are
anti-static. While we can find plenty of anti-static, plastic tweezers
hers
are curved and our searches keep turning up straight or angled pairs. On
one
side is the code "J201" and the flip side has a sort of logo. It looks
like a
J surrounded by four "pine trees". Any assistance would be greatly
appreciated.

Thanks,
Angie




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5, 14 -- Date: Thu, 10 Nov 2005 10:39:55 -0500
5, 14 -- From: "Anjeanette Ormonde" {Anjeanette.Ormonde-at-unilever.com}
5, 14 -- Subject: tweezers search
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==============================Original Headers==============================
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From: Willem.Wennekes-at-comcast.net
Date: Thu, 10 Nov 2005 17:39:55 -0600
Subject: [Microscopy] AskAMicroscopist: Dry pump emission

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This Question was submitted to Ask-A-Microscopist by (Willem.Wennekes-at-comcast.net)
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Email: Willem.Wennekes-at-comcast.net
Name: Willem Wennekes

Organization: UES

Education: Graduate College

Location: Dayton, OH, USA

Title: Dry pump emission

Question: Hi, We need to build an exhaust system for our oil pumps. The reason is the carcinogenic nature of the pump oil and therefore the exhaust. Since these pumps are used as backing pumps for systems that do not emit any other hazardous gazes, replacing these pumps with compact dry pumps seems to be a good alternative. However, scroll pumps emit graphite and we do not want to have that in the atmosphere either. I like a piston pump made by Leybold, the Ecodry M, which is advertised to have very low particle emission. Does anybody has experience with this pump or has a suggestion for a zero emission dry pump.

Thanks,

Willem Wennekes

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From: foxglovelj-at-msn.com
Date: Thu, 10 Nov 2005 17:40:39 -0600
Subject: [Microscopy] AskAMicroscopist: 10 year old son who wants a microscope for xmas

Contents Retrieved from Microscopy Listserver Archives
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This Question was submitted to Ask-A-Microscopist by (foxglovelj-at-msn.com)
from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, November 10, 2005 at 14:57:05
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Email: foxglovelj-at-msn.com
Name: Laura Giard

Education: Undergraduate College

Location: Sterling, MA USA

Title: microscope purchase

Question: Hello, I have a 10 year old son who wants a microscope for xmas. He wants to be a scientist when he grow up! We have purchased "cheap" microscopes over the years and have been very disappointed. I'd like to find one that truly works, however, cannot spend a fortune. Can you give me some help? Magnifications etc. do not really help me in determining the quality which is the only information I can seem to gain from places that sell them. Right now I've been looking at ones through Nasco. ANY HELP would be GREATLY APPRECIATED! Thank you.

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From: diller-at-stefan-diller.com
Date: Thu, 10 Nov 2005 17:43:34 -0600
Subject: [Microscopy] viaWWW: KEVEX DELTA plus analyzer

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Email: diller-at-stefan-diller.com
Name: Stefan Diller

Organization: Photography

Title-Subject: [Filtered] MListserver:

Question: Dear all,
does anybody have experience and / or programms to interface a KEVEX DELTA plus analyzer to a modern PC via RS232?
My modell runs the Quantex + software on two Bernoulli 44MB drives.
I have a second Bernoulli set available.
If there is no other soultion, is there a possibility to get the Bernoulli working via SCSI on a PC to exchange the specs / data?

Thanks,
Stefan Diller

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

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From: gary-at-gaugler.com
Date: Thu, 10 Nov 2005 17:50:22 -0600
Subject: [Microscopy] Re: AskAMicroscopist: Dry pump emission

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

I would suggest the Edwards XDS5 or XDS10 dry scroll pump
with a BOC Edwards XDS Silencer A50597000. The silencer
is a filter trap on the output. it is available for about
$200. I use both of these pumps and have no problems.

gary g.


At 03:42 PM 11/10/2005, you wrote:



} ----------------------------------------------------------------------------
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From: r.sims-at-auckland.ac.nz
Date: Thu, 10 Nov 2005 18:04:59 -0600
Subject: [Microscopy] re: Dry pump emission

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

Date sent: Thu, 10 Nov 2005 17:41:26 -0600
To: r.sims-at-auckland.ac.nz
X-from: Willem.Wennekes-at-comcast.net
}
} Question: Hi, We need to build an exhaust system for our oil pumps.
} The reason is the carcinogenic nature of the pump oil and therefore
} the exhaust. Since these pumps are used as backing pumps for systems
} that do not emit any other hazardous gazes, replacing these pumps with
} compact dry pumps seems to be a good alternative. However, scroll
} pumps emit graphite and we do not want to have that in the atmosphere
} either. I like a piston pump made by Leybold, the Ecodry M, which is
} advertised to have very low particle emission. Does anybody has
} experience with this pump or has a suggestion for a zero emission dry
} pump.
}


Are you sure the exhaust vapor from a rotary pump is carcinogenic?

Granted, it may not be pleasant or healthy, but as I understand it these oils are just
aliphatic hydrocarbon mineral oils, and they're not carcinogenic, are they?

An oil mist trap can clean up the exhaust to the stage where it will bother no-one,
particularly if discharged to out-of-doors.

Any rotary-pump-oil manufacturers listening? Any well-informed (I'm not) comment on
this?

cheers

rtch

--
Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713
Microanalyst Fax : 64 9 3737435
Department of Geology email : r.sims-at-auckland.ac.nz
The University of Auckland
Private Bag 92019
Auckland
New Zealand


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From: schooley-at-mcn.org
Date: Thu, 10 Nov 2005 18:24:27 -0600
Subject: [Microscopy] Re: AskAMicroscopist: 10 year old son who wants a

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

} Question: Hello, I have a 10 year old son who wants a microscope
} for xmas. He wants to be a scientist when he grow up! We have
} purchased "cheap" microscopes over the years and have been very
} disappointed. I'd like to find one that truly works, however,
} cannot spend a fortune. Can you give me some help? Magnifications
} etc. do not really help me in determining the quality which is the
} only information I can seem to gain from places that sell them.
} Right now I've been looking at ones through Nasco. ANY HELP would
} be GREATLY APPRECIATED! Thank you.
}
I urge you to look at the advice about buying microscopes on the
MICRO website - URL below. You can start with MICRO's recommended
20x monocular "dissecting scope" (about $75) and go on to a basic
compound scope (about $150) if he is really interested; both are
needed if he's serious. Order the inspirational "Hidden Worlds -
Looking Through a Scientist's Microscope" (less than $5 online), and
one of the several books about simple specimen preparation listed in
the MICRO bibliography, when you get the compound scope. And Email
me directly if you have further questions.

Caroline

--
Caroline Schooley
Project MICRO Coordinator
Microscopy Society of America
Box 117, 45301 Caspar Point Road
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.microscopy.org/ProjectMICRO

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From: tivol-at-caltech.edu
Date: Thu, 10 Nov 2005 18:32:23 -0600
Subject: [Microscopy] Re: AskAMicroscopist: 10 year old son who wants a microscope for xmas

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


On Nov 10, 2005, at 3:40 PM, foxglovelj-at-msn.com wrote:

} Email: foxglovelj-at-msn.com
} Name: Laura Giard
}
} Education: Undergraduate College
}
} Location: Sterling, MA USA
}
} Title: microscope purchase
}
} Question: Hello, I have a 10 year old son who wants a microscope for
} xmas. He wants to be a scientist when he grow up! We have purchased
} "cheap" microscopes over the years and have been very disappointed.
} I'd like to find one that truly works, however, cannot spend a
} fortune. Can you give me some help? Magnifications etc. do not
} really help me in determining the quality which is the only
} information I can seem to gain from places that sell them. Right now
} I've been looking at ones through Nasco. ANY HELP would be GREATLY
} APPRECIATED! Thank you.
}
Dear Laura,
Check out Project MICRO; go to the MSA web site
(www.msa.microscopy.org), click on the Reference & Educational button
on the left side, and then on ProjectMICRO, the 6th item under
Education Committee Functions.
Yours,
Bill Tivol, PhD
EM Scientist and Manager
Cryo-Electron Microscopy Facility
Broad Center, Mail Code 114-96
California Institute of Technology
Pasadena CA 91125
(626) 395-8833
tivol-at-caltech.edu


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From: bozzola-at-siu.edu
Date: Thu, 10 Nov 2005 18:48:59 -0600
Subject: [Microscopy] re: carcinogenicity of dry pump emissions

Contents Retrieved from Microscopy Listserver Archives
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Good question. It is generally accepted that long term irritants
(even at low levels) can be carcinogenic. For example, a jagged edge
of a tooth can cause an oral cancer due to constant irritation. Oil
mists (while not carcinogenic per se) can irritate the lungs and
cause pneumonia or lung inflammation. Organo-metallics (and perhaps
by extension, used pump oils) are known carcinogens. Used motor oil,
for example, since it contains organo-metallics, is considered a low
level carcinogen. I am uncertain (but suspicious, however) if used
synthetic oils are similarly carcinogenic.

Microscopists can spend many hours in a room with a vacuum pump. Over
a period of years, well ...... let's just say that it's worrisome. In
our facility, we exhaust the oil mist from our pumps outside the
building where it condenses on the roof of the building and is
ultimately washed by rain into the soil. Not ideal, but the oil is
diluted by water and can be broken down by microbes over time rather
than being concentrated in one's lungs.

Many years ago, I evaluated an exhaust filter that trapped over 99+%
of the oil mist and returned it to the pump. However, after I placed
a Petri dish with distilled water in the room, I could observe (by
eye as well as under a stereomicroscope) tiny droplets of oil landing
on the water surface and spreading out to form an "oil slick". That's
when I decided to exhaust to the outside.

Maybe the risk of lung cancer is low but why take the chance?

Cheers,

John B.


} Date sent: Thu, 10 Nov 2005 17:41:26 -0600
} To: r.sims-at-auckland.ac.nz
} X-from: Willem.Wennekes-at-comcast.net
} }
} } Question: Hi, We need to build an exhaust system for our oil pumps.
} } The reason is the carcinogenic nature of the pump oil and therefore
} } the exhaust. Since these pumps are used as backing pumps for systems
} } that do not emit any other hazardous gazes, replacing these pumps with
} } compact dry pumps seems to be a good alternative. However, scroll
} } pumps emit graphite and we do not want to have that in the atmosphere
} } either. I like a piston pump made by Leybold, the Ecodry M, which is
} } advertised to have very low particle emission. Does anybody has
} } experience with this pump or has a suggestion for a zero emission dry
} } pump.
} }
}
}
} Are you sure the exhaust vapor from a rotary pump is carcinogenic?
}
} Granted, it may not be pleasant or healthy, but as I understand it
} these oils are just
} aliphatic hydrocarbon mineral oils, and they're not carcinogenic, are they?
}
} An oil mist trap can clean up the exhaust to the stage where it will
} bother no-one,
} particularly if discharged to out-of-doors.
}
} Any rotary-pump-oil manufacturers listening? Any well-informed (I'm
} not) comment on
} this?
}
} cheers
}
} rtch
}
} --
} Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713
} Microanalyst Fax : 64 9 3737435
} Department of Geology email : r.sims-at-auckland.ac.nz
} The University of Auckland
} Private Bag 92019
} Auckland
} New Zealand
}
}
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--
##############################################################
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
750 Communications Drive - MC 4402
Southern Illinois University
Carbondale, IL 62901 U.S.A.
Phone: 618-453-3730
Email: bozzola-at-siu.edu
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##############################################################

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From: gary-at-gaugler.com
Date: Thu, 10 Nov 2005 18:56:51 -0600
Subject: [Microscopy] Re: AskAMicroscopist: 10 year old son who wants a

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

You can find some very nice scopes on ebay. Try
for Olympus BH-2/BHT. Look for a unit with trinoc
port and 50W or 100W lamp house. These scopes use
legacy 160mm tube length objectives and can work with
Oly, Nikon, Zeiss, etc. objectives. Over time, you
can work up from Plan objectives to PlanAPO and add
phase and DIC (hard to find). The Zeiss 63X PlanAPO
was probably one of the best 160 objectives they made,
IMO.

This all assumes that you want a compound LM. If you
want a stereo, then look for Oly SH series scopes
or Nikons. These were used extensively in semiconductor
inspection and show up used routinely. But be sure
that the scope has a trinoc port if you ever want to
be able to take pix the best way (versus an ocular).

gary g.


At 03:42 PM 11/10/2005, you wrote:



} ----------------------------------------------------------------------------
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From: christopher.hayden-at-novartis.com
Date: Fri, 11 Nov 2005 07:54:43 -0600
Subject: [Microscopy] viaWWW: Eye Fixation Protocol Thank-you

Contents Retrieved from Microscopy Listserver Archives
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Yes, sure, we want to avoid irritants, but I still would be careful about the "c" word, it
gets tagged on to things so easily, and sticks like glue.

As you say, its the organometallics that make used motor oil a low-level carcinogen,
not the base oil, and good rotary pump oils are just very pure base oils, aren't they?

And who uses synthetic oils in their rotary pumps?

Please note that I'm not advocating exposure to pump exhausts, its just the possibly
careless use of "carcinogenic" that I mind.

cheers

rtch


Date sent: Thu, 10 Nov 2005 18:50:15 -0600
To: r.sims-at-auckland.ac.nz
X-from: bozzola-at-siu.edu
Send reply to: bozzola-at-siu.edu

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html
---------------------------------------------------------------------------
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Email: christopher.hayden-at-novartis.com
Name: Christopher Hayden

Organization: Novartis Pharmaceuticals

Title-Subject: [Filtered] Eye Fixation Protocol Thank-you

Question: Good Morning, All:

A sincere (and belated) thank-you to everyone who responded to my question about the fixation of mammalian eyes. We ended up going with 6% glutaraldehyde for fixation (one of the many suggested procedures). I'd love to tell everyone how it came out, but our priorities have shifted and it'll be a while before we get to actually scope the samples.

I wanted to write back to everyone indivially to thank them, but due some electronic problems I lost nearly all of my saved email(!), so it's been a fun few weeks trying to get back to "normal". We couldn't have done it as well without everyone's help.

My thanks.
-Chris

--------------
Christopher Hayden
PCS/Electron Microscopy
Novartis Pharmaceuticals
christopher.hayden-at-novartis.com

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From: simon.ringer-at-emu.usyd.edu.au
Date: Fri, 11 Nov 2005 07:58:07 -0600
Subject: [Microscopy] viaWWW: ACMM19 Conference Deadlines Approaching

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Email: simon.ringer-at-emu.usyd.edu.au
Name: Simon Ringer

Organization: University of Sydney

Title-Subject: [Filtered] MListserver: ACMM19

Question: Dear colleague,

I am taking the liberty of contacting you to highlight that the 19th Australian Conference for Microscopy and Microanalysis (ACMM19) will be held in Sydney, Australia, on the 5th to 9th February 2006. The biennial ACMM meetings are one of Australiaís most important forums for the microscopy and microanalytical sciences, covering the gamut of techniques and applications, and attracting a significant international attendance. More information on ACMM19 may be found on the conference web site at

http://www.acmm19.org.au.

I will take this opportunity, however, to draw your attention specifically to three points:
(1) late submissions of abstracts for the conference will be accepted for a few more days;
(2) a late-breaking poster session will be held at the conference; and
(3) the early bird registration closes December 5th.

I, and the rest of the organising committee, look forward to a stimulating scientific interaction with you at ACMM19 in February.

Regards,
Simon

Professor Simon P. Ringer
Convenor, ACMM19

Director
Australian Key Centre for Microscopy & Microanalysis

Executive Director
Nanostructural Analysis Network Organisation
Major National Research Facility
NANO-MNRF
www.nano.org.au

Address:
Electron Microscope Unit
The University of Sydney
NSW, 2006
AUSTRALIA

Contact:
PH) + 61 2 9351 2351
FAX) + 61 2 9351 7682
simon.ringer-at-emu.usyd.edu.au
www.emu.usyd.edu.au

Madsen Building, F09
Room LG21
CRICOS Number: 00026A www.usyd.edu.au/disclaimer.shtml


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From: wesaia-at-iastate.edu
Date: Fri, 11 Nov 2005 08:58:03 -0600
Subject: [Microscopy] Re: viaWWW: KEVEX DELTA plus analyzer

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

Stefan, I have done this in the past, but it might take me a while to dig
out the details. I can give you a quick synopsis now and we can talk more
later.

I have used serial and ethernet communication to transfer data between the
Delta and the rest of the world. I think I tried hooking up a Bernoulli
drive, but then you need a utility that can read the directory under
DOS/Windows. It is not a trivial thing. I received a copy of one, but I
don't know that I ever got it to work.

Serial transfer should work with what you have now. You would only need a
cable to plug between the two systems and maybe some software tweaks. I
don't remember if the Kevex RT-11/TSX+ operating system came with a driver
built in for the serial port. I think it had a driver for RT-11; I don't
quite remember about TSX+. I have rebuilt copies of the operating system
years ago to add different drivers, but it has been a while. Let's suppose
the drivers are built in, the cable should be fairly straightforward. You
may want to find someone with a breakout box to make sure the pins are all
setup correctly. The Kevex doesn't hardware handshaking lines, but I think
the PC does. Finally, you need some software on both ends. I used Kermit
which was a rather ubiquitous program written for lots of platforms. I will
have to see if I still have copies around. It supported both text and
binary transfer. One end should be set up in server mode so the other
computer can push or pull files around.

We went with an ethernet option because serial is slow. I purchased an
ethernet card and software that allowed me to put the PDP onto an ethernet
network. I think the software may only have worked under TSX, but that was
okay because we usually ran under TSX than RT-11 anyway. I think I had to
rebuild the operating system to add in the capability. It might be more
daunting.

The next and primary question will be what will you do with the files once
you have transferred them over? A lot of Kevex stuff was in a proprietary
binary format. I developed procedures for transferring text output.
However, spectra and images are another story. I think Kevex may have
offered a spectrum export option. I developed my own utilities for
converting maps and images over to TIF or BMP format.

So before spending much time hooking the two systems together, what do you
want to do with the files once you can move them?

Warren

At 05:44 PM 11/10/05, you wrote:


} Email: diller-at-stefan-diller.com
} Name: Stefan Diller
}
} Organization: Photography
}
} Title-Subject: [Filtered] MListserver:
}
} Question: Dear all,
} does anybody have experience and / or programms to interface a KEVEX DELTA
} plus analyzer to a modern PC via RS232?
} My modell runs the Quantex + software on two Bernoulli 44MB drives.
} I have a second Bernoulli set available.
} If there is no other soultion, is there a possibility to get the Bernoulli
} working via SCSI on a PC to exchange the specs / data?
}
} Thanks,
} Stefan Diller


==============================Original Headers==============================
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From: Rob.Bowen-at-caddock.com
Date: Fri, 11 Nov 2005 10:02:03 -0600
Subject: [Microscopy] Re: viaWWW: KEVEX DELTA plus analyzer

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

Stefan,
Look at IXRF, http://www.ixrfsystems.com/index.html . They offer
upgrades for Kevex systems and may have something you can use.

No interest in IXRF except as a satisfied user.

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

} From: {diller-at-stefan-diller.com}
} Reply-To: {diller-at-stefan-diller.com}
} Date: Thu, 10 Nov 2005 17:46:35 -0600
} To: {rob.bowen-at-caddock.com}
} Subject: [Microscopy] viaWWW: KEVEX DELTA plus analyzer
}
}
}
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} Email: diller-at-stefan-diller.com
} Name: Stefan Diller
}
} Organization: Photography
}
} Title-Subject: [Filtered] MListserver:
}
} Question: Dear all,
} does anybody have experience and / or programms to interface a KEVEX DELTA
} plus analyzer to a modern PC via RS232?
} My modell runs the Quantex + software on two Bernoulli 44MB drives.
} I have a second Bernoulli set available.
} If there is no other soultion, is there a possibility to get the Bernoulli
} working via SCSI on a PC to exchange the specs / data?
}
} Thanks,
} Stefan Diller
}
} ---------------------------------------------------------------------------
}
} ==============================Original Headers==============================
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From: eschumacher-at-mccrone.com
Date: Fri, 11 Nov 2005 11:22:22 -0600
Subject: [Microscopy] Electron Microscope Identification

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

Greetings All,

A colleague here has been asked whether we can reproduce work outlined
in a rather
vaguely worded document that describes characterization of hydrated
sections of
film. Photomicrographs are shown that are said to have been acquired
using a
"DEH 345" penetrating electronic microscope. The data bar on the
photomicrographs
has a 6-digit number in the upper left corner that's preceded by the
letters "ES".
We wondered if this might refer to an Electroscan environmental SEM.
Our searches
on the instrument description haven't turned up anything. I'm hoping
that the
collective expertise of the listserver will shed some light on this for
us!

Thank you,

Elaine


Elaine F. Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com




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From: jrunions-at-brookes.ac.uk
Date: Fri, 11 Nov 2005 11:40:40 -0600
Subject: [Microscopy] Planning a Masters course in Bioimaging

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


Hi Everyone (Sorry if you've already seen this on the confocal
microscopy list),

We are in the planning and validation stage of a Masters in Bioimaging
program to be offered at Oxford Brookes University starting in Sep.
2006. This course is supported by the Royal Microscopical Society and
we have gotten good feedback from industry regarding support. I am
seeking input from others who have been involved in or are contemplating
such a course. As part of the validation before the University
committee, we need to show that such a course is viable and that there
would be a reasonable demand for it. The University is doing away with
those smaller Masters programs that only attract a few students per
year. My feeling is that this is no problem with the renaissance in
imaging that confocal microscopy has produced but I need something more
concrete to present to the committee.

Specifically:

1) Are there other Graduate level courses in Bioimaging in the UK or
internationally? If so, how are they doing?
2) What do you feel is the best way to advertise the course, both to new
graduates and to those in industry who might like to take a module or two?
3) If you are a professional in an imaging-related company, what would
you be interested in learning as a module in such a course?

I really want to hear from you whether your views are positive or
otherwise. If you are in the UK and you would like to be involved as a
guest lecturer, sponsor etc., by all means let me know.

Thank you.

Sincerely, John.
--

*********************************
C. John Runions, Ph.D.
School of Biological and Molecular Sciences
Oxford Brookes University
Oxford, UK
OX3 0BP

email: jrunions-at-brookes.ac.uk
phone: +44 (0) 1865 483 964


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From: bozzola-at-siu.edu
Date: Fri, 11 Nov 2005 16:56:38 -0600
Subject: [Microscopy] Refs: carcinogenicity of used pump oil

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

Used motor oil is considered carcinogenic (see 1 and 2, below).

It appears that short term exposure to oil fumes may not produce
significant adverse health effects according to CITGO (see 2, below).

I can send the full references if you like. Sorry, I didn't have the
time to reformat the quotes. I'll see if I can find any better refs.

Some quotes are enclosed.

1. Brief Summary of Carcinogenicity/Cancer Information:
The 4- to 7-ring PAHs have been especially implicated in the
carcinogenic effect of used oil [519]. Many of these same PAHs are
found in new oil, although in lower concentrations [519]. Above text
reprinted with permission from Environmental Toxicology and
Chemistry, Volume 12(11), C. Upshall, J.F. Payne, and J. Hellou,
"Induction of MFO Enzymes and Production of Bile Metabolites in
Rainbow Trout (Oncorhynchus mykiss) Exposed to Waste Crankcase Oil."
Copyright 1993 SETAC].

The debates on which PAHs and alkyl PAHs in complex mixtures such as
this product to classify as carcinogens, and the details of exactly
how to perform both ecological and human risk assessments on the
complex mixtures of PAHs typically found at contaminated sites, are
likely to continue. There are some clearly wrong ways to go about
it, but defining clearly right ways is more difficult. PAHs usually
occur in complex mixtures rather than alone.

Perhaps the most unambiguous thing that can be said about complex PAH
mixtures is that such mixtures are often carcinogenic and possibly
phototoxic. One way to approach site specific risk assessments would
be to collect the complex mixture of PAHs and other lipophilic
contaminants in a semipermeable membrane device (SPMD, also known as
a fat bag) [894,895,896], retrieve the contaminant mixture from the
SPMD, then test the mixture for carcinogenicity, toxicity, and
phototoxicity (James Huckins, National Biological Service, and Roy
Irwin, National Park Service, personal communication, 1996).
Source: ENVIRONMENTAL CONTAMINANTS ENCYCLOPEDIA OIL, NEW (UNUSED
MOTOR OIL) ENTRY July 1, 1997 COMPILERS/EDITORS: ROY J. IRWIN,
NATIONAL PARK SERVICE WITH ASSISTANCE FROM COLORADO STATE UNIVERSITY
STUDENT ASSISTANT CONTAMINANTS SPECIALISTS: MARK VAN MOUWERIK LYNETTE
STEVENS MARION DUBLER SEESE WENDY BASHAM NATIONAL PARK SERVICE WATER
RESOURCES DIVISIONS, WATER OPERATIONS BRANCH 1201 Oakridge Drive,
Suite 250 FORT COLLINS, COLORADO 80525

2. From the CITGO MSDS (material safety data sheet):
SECTION 3: HAZARDS IDENTIFICATION
Inhalation No significant adverse health effects are expected to
occur upon short-term exposure.
CAUTION: Hot oil can cause thermal burns on contact. "Used" motor oil
has been associated with skin cancer in laboratory animals following
extended contact.

3. From SYNLUBE.COM:
Used Motor Oil, if handled improperly and without proper personal
protection and hygiene is proven to be cancer causing.
Look at any back label of Petroleum or Synthetic Motor Oil made after
1985, and you will find following statement:
"CAUTION: Avoid prolonged or repeated skin contact with used motor
oil. Used motor oil has been shown to cause skin cancer in laboratory
animals. Thoroughly wash exposed areas with soap and water."


--
##############################################################
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
750 Communications Drive - MC 4402
Southern Illinois University
Carbondale, IL 62901 U.S.A.
Phone: 618-453-3730
Email: bozzola-at-siu.edu
Web: http://www.siu.edu/~image/
##############################################################

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From: sghoshro-at-NMSU.Edu
Date: Fri, 11 Nov 2005 17:29:54 -0600
Subject: [Microscopy] Thank you

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


Hi John

Thanks for your detailed reply.

I would be surprised, though, if rotary pump oil, new or used, contains any PAHs, and it
still seems to me that to extend the established carcinogenicity of used engine oil to
rotary vacuum pump oil is a bit of a stretch, isn't it?

Of course it is obviously best to play safe with exposure to any chemical products, but I
don't think we should label something as carcinogenic without fairly good cause.

cheers

rtch


Date sent: Fri, 11 Nov 2005 16:58:07 -0600
To: r.sims-at-auckland.ac.nz
X-from: bozzola-at-siu.edu
Send reply to: bozzola-at-siu.edu

Thanks to everyone who responded to my inquiry regarding fixation of
Hepatitis C infected tissue. We revceived some great responses.

Best wishes,

Soumitra


*************************************************************
Soumitra Ghoshroy
College Associate Professor, Biology
Director, Electron Microscopy Lab
Box 3EML
New Mexico State University
Las Cruces, NM 88003
Tel: 505-646-3268 (office), 646-3283 (lab)
Fax: 505-646-3282
e-mail:sghoshro-at-nmsu.edu
http://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm
http://emldata.nmsu.edu/eml/

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From: r.sims-at-auckland.ac.nz
Date: Fri, 11 Nov 2005 17:34:18 -0600
Subject: [Microscopy] Regenation of Al2O3 granules

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

Hi

Presumably the Al2O3 granules used in Edwards and other foreline traps can be
regenerated by heating in a furnace.

What temperature and time is necessary to bake/burn off trapped oil without destroying
the physical structure so that Al2O3 dust goes into the rotary pump?

cheers

rtch

--
Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713
Microanalyst Fax : 64 9 3737435
Department of Geology email : r.sims-at-auckland.ac.nz
The University of Auckland
Private Bag 92019
Auckland
New Zealand


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From: jtwilley-at-sprynet.com
Date: Fri, 11 Nov 2005 20:10:14 -0600
Subject: [Microscopy] Refs: carcinogenicity of used pump oil

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


Of course there's no way we can make alumina into royalty, I meant "Regeneration"

cheers

rtch


------- Forwarded message follows -------
Date sent: Fri, 11 Nov 2005 17:35:00 -0600
To: r.sims-at-auckland.ac.nz
X-from: r.sims-at-auckland.ac.nz
Send reply to: r.sims-at-auckland.ac.nz

There seems to be a leap from one subject to another here. Used motor oil is just that, motor oil that was used in an internal combustion engine where a thin film of the oil with a very high surface area has been repeatedly exposed to the high temp, high pressure combustion conditions inside the cylinders. That oil is exposed to transitory, highly reactive species and highly poisonous species including free radicals, nitrogen oxides, carbon monoxide, and a host of partially oxidized hydrocarbons and water vapor. Catalytic converters are used downstream to destroy the more troublesome persistent species from the exhaust but the oil absorbs some of them and cycles them back through the cylinders over and over. These species are also free to react with finely divided metal particles and detergent additives in the oil. Its an environment where a plethora of new species are created and, once created, allowed to react with each other. The high pressure also influences the situation by increasing the tendency to force gaseous species into solution in the oil, prolonging their opportunities to react, rather than encouraging their vapor phase dilution.

Whether any chemistry is going on at all in vacuum pump oil depends on what is being exhausted. But it's safe to say that many lab pumps are not run under conditions that make them a pressure reactor like the one that exists in every internal combustion engine.

John Twilley

-----Original Message-----
X-from: bozzola-at-siu.edu
Sent: Nov 11, 2005 5:57 PM
To: jtwilley-at-sprynet.com

Used motor oil is considered carcinogenic (see 1 and 2, below).

It appears that short term exposure to oil fumes may not produce
significant adverse health effects according to CITGO (see 2, below).

I can send the full references if you like. Sorry, I didn't have the
time to reformat the quotes. I'll see if I can find any better refs.

Some quotes are enclosed.

1. Brief Summary of Carcinogenicity/Cancer Information:
The 4- to 7-ring PAHs have been especially implicated in the
carcinogenic effect of used oil [519]. Many of these same PAHs are
found in new oil, although in lower concentrations [519]. Above text
reprinted with permission from Environmental Toxicology and
Chemistry, Volume 12(11), C. Upshall, J.F. Payne, and J. Hellou,
"Induction of MFO Enzymes and Production of Bile Metabolites in
Rainbow Trout (Oncorhynchus mykiss) Exposed to Waste Crankcase Oil."
Copyright 1993 SETAC].

The debates on which PAHs and alkyl PAHs in complex mixtures such as
this product to classify as carcinogens, and the details of exactly
how to perform both ecological and human risk assessments on the
complex mixtures of PAHs typically found at contaminated sites, are
likely to continue. There are some clearly wrong ways to go about
it, but defining clearly right ways is more difficult. PAHs usually
occur in complex mixtures rather than alone.

Perhaps the most unambiguous thing that can be said about complex PAH
mixtures is that such mixtures are often carcinogenic and possibly
phototoxic. One way to approach site specific risk assessments would
be to collect the complex mixture of PAHs and other lipophilic
contaminants in a semipermeable membrane device (SPMD, also known as
a fat bag) [894,895,896], retrieve the contaminant mixture from the
SPMD, then test the mixture for carcinogenicity, toxicity, and
phototoxicity (James Huckins, National Biological Service, and Roy
Irwin, National Park Service, personal communication, 1996).
Source: ENVIRONMENTAL CONTAMINANTS ENCYCLOPEDIA OIL, NEW (UNUSED
MOTOR OIL) ENTRY July 1, 1997 COMPILERS/EDITORS: ROY J. IRWIN,
NATIONAL PARK SERVICE WITH ASSISTANCE FROM COLORADO STATE UNIVERSITY
STUDENT ASSISTANT CONTAMINANTS SPECIALISTS: MARK VAN MOUWERIK LYNETTE
STEVENS MARION DUBLER SEESE WENDY BASHAM NATIONAL PARK SERVICE WATER
RESOURCES DIVISIONS, WATER OPERATIONS BRANCH 1201 Oakridge Drive,
Suite 250 FORT COLLINS, COLORADO 80525

2. From the CITGO MSDS (material safety data sheet):
SECTION 3: HAZARDS IDENTIFICATION
Inhalation No significant adverse health effects are expected to
occur upon short-term exposure.
CAUTION: Hot oil can cause thermal burns on contact. "Used" motor oil
has been associated with skin cancer in laboratory animals following
extended contact.

3. From SYNLUBE.COM:
Used Motor Oil, if handled improperly and without proper personal
protection and hygiene is proven to be cancer causing.
Look at any back label of Petroleum or Synthetic Motor Oil made after
1985, and you will find following statement:
"CAUTION: Avoid prolonged or repeated skin contact with used motor
oil. Used motor oil has been shown to cause skin cancer in laboratory
animals. Thoroughly wash exposed areas with soap and water."


--
##############################################################
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
750 Communications Drive - MC 4402
Southern Illinois University
Carbondale, IL 62901 U.S.A.
Phone: 618-453-3730
Email: bozzola-at-siu.edu
Web: http://www.siu.edu/~image/
##############################################################

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From: zaluzec-at-microscopy.com
Date: Sat, 12 Nov 2005 08:44:03 -0600
Subject: [Microscopy] Safety/Carcinogenicity of materials

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

Colleagues

I'll just point out that a good STARTING point when ever you are unsure and
want to find out the generic hazards of a material are the Materials Safety Data Sheets (MSDS).

These are required by virtually all safety departments in most organziations and are
also usually shipped with materials or are available on-line from the manufacturer.

You can also MSDSs on-line at various WWW sites here is a list of
starting points. In general it is a good policy to have access to data sheets
on all chemcials you use in your lab.

http://www.ilpi.com/msds/#What


Most of the sites listed above are free, but a few require registration.

FYI appended is a short excerpt on "Edwards High Vacuum Mechanical Pump Oil"
which is a pretty common mechanical pump oil around EM Labs. You will notice
it says this particular material is not carcinogenitic, although it does not discuss "used" oil.
It does recommend avoiding the mist, which is a good policy.

Having said this, I will concur with others that venting roughing pump systems to
areas outside the immediate laboratory area is generally a good idea. For the most part we
exhaust roughing pumps using simple PVC piping into fume hoods (some of which are
a good distance away ~ 100 ft) the exhaust of those hoods are filtered
and and NOT released into the immediate laboratory air. I prefer this to simple
mist filters that are mounted on the exhaust port of the pumps. Admittedly this
costs more, but it is remember that you or your colleague may end up spending "years"
in a lab and it makes more sense to be cautious rather than cheap.



Nestor
Your Friendly Neighborhood SysOp







EDWARDS HIGH VACUUM INTERNATIO -- H026-00-008 EDWARDS 16 MECHANICAL PUMP OIL
=======================================================
MSDS Safety Information
=======================================================
FSC: 9150
MSDS Date: 08/01/1996
MSDS Num: CHZKX
LIIN: 00F025284
Product ID: H026-00-008 EDWARDS 16 MECHANICAL PUMP OIL

=======================================================
Health Hazards Data
=======================================================
LD50 LC50 Mixture: ORAL LD50(RAT): } 5 ML/KG
Route Of Entry Inds - Inhalation: YES
Skin: NO
Ingestion: YES
Carcinogenicity Inds - NTP: NO
IARC: NO
OSHA: NO
Effects of Exposure: SKIN: DERMATITIS & ERYTHEMA. EYES: IRRITATION OF THE
CONJUNCTIVA. INHALATION: MAY CAUSE A CHRONIC INFLAMMATORY REACTION OF THE
LUNGS & A FORM OF PULMONARY FIBROSIS. INGESTION: ASPIRATION OF LIQUID INT
O THE LUNGS COULD CAUSE PNEUMONIA.
Explanation Of Carcinogenicity: NONE
Signs And Symptions Of Overexposure: DEFATTING, OIL ACNE, OIL FOLICULITIS,
IRRITATION.
First Aid: SKIN/EYES: WASH W/COPIOUS AMOUNTS OF WATER FOR 10 MINS. INHALATION:
REMOVE TO FRESH AIR. INGESTION: DON'T INDUCE VOMITING. SEND TO HOSPITAL
IMMEDIATELY. OBTAIN MEDICAL ATTENTION IN ALL CASES.
=======================================================
Handling and Disposal
=======================================================
Spill Release Procedures: PREVENT ENTRY TO DRAINS & WATERCOURSES. LARGE:
USE A SUITABLE MEDIUM SUCH AS SAND/EARTH. RECLAIM LIQUID DIRECTLY/IN AN
ABSORBENT MEDIUM THEN TRANSFER TO SUITABLE MARKED CONTAINERS & DISPOSE
OF. SMALL: SOAK UP W/SAND/EARTH & DISPOSE OF.
Waste Disposal Methods: DISPOSE OF TO A LICENSED WASTE CONTRACTOR, IAW/FEDERAL,
STATE & LOCAL REGULATIONS.
Handling And Storage Precautions: OIL MIST SHOULD BE KEPT TO A MINIMUM,
PREFERABLY WELL {5 MG/CUM. STORE AWAY FROM DIRECT HEAT.
Other Precautions: AVOID OIL MIST FUMES & VAPORS.
=======================================================
Fire and Explosion Hazard Information
=======================================================
Flash Point Method: PMCC
Flash Point Text: 460.4F
Extinguishing Media: FOAM, DRY POWDER, CO2, HALON.
Fire Fighting Procedures: DON'T USE WATER JETS. WEAR A SCBA IN CONFINED AREAS
& FOR ANY SIGNIFICANT FIRE.
Unusual Fire/Explosion Hazard: AUTOIGNITION TEMP: 608F. THERE IS A POSSIBILITY
OF EXPLOSION IF YOU PUMP GASES CONTAINING OXYGEN AT A CONCENTRATION } 4%
ABOVE THE PROPORTION CONTAINED IN AIR.
=======================================================
Control Measures
=======================================================
Respiratory Protection: NOT REQUIRED UNDER NORMAL CONDITIONS.
Ventilation: LOCAL EXHAUST W/MIST FILTER, EXHAUST TO ATMOSPHERE.
Protective Gloves: IMPERVIOUS POLY VINYL CHLORIDE
Eye Protection: NOT NECESSARY
Other Protective Equipment: OVERALLS.



==================================================================================



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From: gary-at-gaugler.com
Date: Sun, 13 Nov 2005 13:17:11 -0600
Subject: [Microscopy] specimen rotation unit

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

Hi Listers:

Does anyone know of a rotatable specimen holder
for "standard" pin stubs? What I would like is
a holder to take 3mm pin stubs and be able to
externally rotate it on the main specimen holder.

The application is that the specimen is on a
45 degree pre-tilt holder. Then, I want to be
able to rotate the specimen using an external joystick
or knob. The unit cannot be too tall or it won't fit
through my specimen load lock.

Any ideas?

gary g.


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From: Colin.Veitch-at-csiro.au
Date: Sun, 13 Nov 2005 17:06:07 -0600
Subject: [Microscopy] Hitachi S4300SE/N information

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

Hi All,

It looks like we'll be getting the funds for a new SEM in the near
future (fingers crossed!!) and we are thinking of going down the field
emission, variable pressure, natural, ESEM route.

I have contacts that are using some of the available systems, but there
are no Hitachi S4300SE/N systems in Australia yet. If anyone out there
has had experience with these machines I would appreciate some feedback
on performance, ease of use etc.. Please feel free to email me
personally with any comments.

Just for the record, we are currently using a Hitachi S4100.

Cheers and thank you very much.

Colin Veitch

Electron Microscopist

CSIRO Textile and Fibre Technology

PO Box 21, BELMONT, Vic. 3216. Australia.

E-mail: colin.veitch-at-csiro.au

Web: http://www.tft.csiro.au

Tel: +61 (0) 3 5246 4000
Mob: 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 Textile and Fibre
Technology on +61 3 5246 4000.



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From: Sally.Stowe-at-anu.edu.au
Date: Sun, 13 Nov 2005 20:05:56 -0600
Subject: [Microscopy] RE: specimen rotation unit

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

I'm after something similar - a two axis motorised tilt plus rotate device,
must tilt to at least 90 degrees and rotate 360, that can be easily and
reveribly attached to the X-Y motorised stage of an SEM.

Doesn't have to be stunningly precise, does have to be cheap, and could be
designed for any of the standard pin or stub mounts.
cheers
Sally


Dr SJ Stowe
Facility Coordinator
ANU Electron Microscopy Unit
ANU CRICOS#00120C



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From: gary-at-gaugler.com
Date: Sun, 13 Nov 2005 21:05:46 -0600
Subject: [Microscopy] specimen rotation unit

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

Deben UK has a tilt and rotate unit that is meant to
work like you say. It is not what I am looking for
but may be for you. The posted unit is for JEOL.

http://www.deben.co.uk/details.php?id=17

As a user/consumer, the Deben products are quite good.

gary g.


At 06:08 PM 11/13/2005, you wrote:



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From: ajheim-at-mail.usf.edu
Date: Sun, 13 Nov 2005 23:16:23 -0600
Subject: [Microscopy] viaWWW: Force microscopy on Asylum MFP-3D

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Email: ajheim-at-mail.usf.edu
Name: A H

Organization: USF

Title-Subject: [Filtered] Force microscopy on Asylum MFP-3D

Question: Is anyone using the AR MFP-3D to perform aqueous force curves on soft or biological materials?



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From: pk-at-aurorasiam.com
Date: Sun, 13 Nov 2005 23:18:29 -0600
Subject: [Microscopy] AskAMicroscopist: What kind of microscope should i get?

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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pk-at-aurorasiam.com) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, November 13, 2005 at 20:30:40
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Email: pk-at-aurorasiam.com
Name: Per Kristian

Organization: Asssumption

Education: Graduate College

Location: Thailand

Question: Hi,

I want to get a microscope. My wish is to watch living organisms like viruses and bacteria to see how they behave.

What kind of microscope should i get?

Best regards,

Per Kristian

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From: hagglundk1-at-nku.edu
Date: Mon, 14 Nov 2005 09:24:34 -0600
Subject: [Microscopy] SEM imaging of bacteria encapsulation

Contents Retrieved from Microscopy Listserver Archives
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We have a student working on a project in which they are growing
bacterial cultures (pseudomonas and other species) in a hostile
environment. The bacteria form capsules that we are interested in
seeing. The capsules are most likely composed of sugars
(polysaccharides), and we are hoping to find a fixation or other
preparation technique that would help us with imaging these. We have so
far prepared cultures with standard fixation, dehydration, and critical
point drying techniques, but are not sure that we are preserving the
capsules forming around the bacteria.

We do not have a cryo stage available.

Any ideas?

Karl Hagglund
Biological Sciences, SC-102B
Northern Kentucky University, Nunn Drive
Highland Heights, KY 41099
859-572-5238
http://semlab.nku.edu


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From: TindallR-at-missouri.edu
Date: Mon, 14 Nov 2005 09:54:13 -0600
Subject: [Microscopy] SEM imaging of bacteria encapsulation

Contents Retrieved from Microscopy Listserver Archives
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You might try using some Alcian blue in the primary fix to stabilise the
capsules.
Works for us for TEM with Campylobacter.

Chris

----- Original Message -----
X-from: {hagglundk1-at-nku.edu}
To: {c.jeffree-at-ed.ac.uk}
Sent: Monday, November 14, 2005 3:25 PM

There are "non-aqueous" fixation techniques and methods involving
ruthenium red and alcian blue that are useful in preserving these
capsules and other biofilms. The first method involves dissolving
osmium tetroxide in a solvent, such as FC-72 from 3M company, rather
than in an aqueous buffer.

For details, see Microscopy Research And Technique 36:390-399 (1997) and
36:422-427 (1997). Also, Biotech Histochem 66(4):173-80 (1991) and J
Comp Pathology 117(2):165-70 (1997).

If you need more references, let me know. I think I have a couple more
somewhere.

Good luck,
Randy

Randy Tindall
EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W122 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





-----Original Message-----
X-from: hagglundk1-at-nku.edu [mailto:hagglundk1-at-nku.edu]
Sent: Monday, November 14, 2005 9:26 AM
To: Tindall, Randy D.

We have a student working on a project in which they are growing
bacterial cultures (pseudomonas and other species) in a hostile
environment. The bacteria form capsules that we are interested in
seeing. The capsules are most likely composed of sugars
(polysaccharides), and we are hoping to find a fixation or other
preparation technique that would help us with imaging these. We have so
far prepared cultures with standard fixation, dehydration, and critical
point drying techniques, but are not sure that we are preserving the
capsules forming around the bacteria.

We do not have a cryo stage available.

Any ideas?

Karl Hagglund
Biological Sciences, SC-102B
Northern Kentucky University, Nunn Drive Highland Heights, KY 41099
859-572-5238
http://semlab.nku.edu


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From: W.Muss-at-salk.at
Date: Mon, 14 Nov 2005 10:11:53 -0600
Subject: [Microscopy] Re: SEM imaging of bacteria encapsulation

Contents Retrieved from Microscopy Listserver Archives
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Hello and Good Morning,

Another proposal:

Tannic acid, LOW molecular weight (that is appr. MW 1600), applied either
in the fixative (0.05 - 0.1 -1.0 %, be sure your glutaraldehyde is of
EM-grade and free of di-, polymers), or fix as you like (buffered as usual,
incl. glutaraldehyde, perhaps higher concentration = {3% to } 6%), wash as
usual, postfix with OsO4 (buffered, 1-2% as usual), start dehydration with
50% EtOH, and immerse then with 1% para-phenylenediamine (PPD, use with
care according to MSDS) in 70% EtOH for at least 30 - 60 min, wash the
specimens several times in pure 70% EtOH (to get rid of non bound PPD) and
go on with dehydration/embedding as usual......

Have not tried acetone as a dehydrating agent, but I don't see a reason why
aceton as a dehydrating agent should fail.....

....... There will be an interesting "difference" in morphology, as
compared to "normal" fixation and dehydration.....(also, you could add a
Tannic acid LOW mol. weight [TA LMW 0.05%-0.1 % hydrous solution, filtered,
finest Millipore or paperfilter) preincubation staining of the ultrathin
sections grids before the classical two-step staining procedure
UO2Ac/Pb-citrate
(TA LMW: 5 min-8 min -at-room temperature, UO2Ac[endvolume 50 ml 1% in EtOH
abs+ adding a drop of acetic acid]15 min, PbCitrate [e.g.
Venable&Coggeshall] 1.5 - 3 min, depending on the freshness of the
Pb-solution)

Best wishes and regards,

Wolfgang Muss
Salzburg, Austria

----------
Von: hagglundk1-at-nku.edu[SMTP:hagglundk1-at-nku.edu]
Antwort an: hagglundk1-at-nku.edu
Gesendet: Montag, 14. November 2005 16:30
An: W.Muss-at-salk.at
Betreff: [Microscopy] SEM imaging of bacteria encapsulation

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We do not have a cryo stage available.

Any ideas?

Karl Hagglund
Biological Sciences, SC-102B
Northern Kentucky University, Nunn Drive
Highland Heights, KY 41099
859-572-5238
http://semlab.nku.edu


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From: heckman-at-bgnet.bgsu.edu
Date: Mon, 14 Nov 2005 10:19:40 -0600
Subject: [Microscopy] Fwd: SEM imaging of bacteria encapsulation

Contents Retrieved from Microscopy Listserver Archives
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Dear Karl-

Saccharides are seen in very high contrast with the use of ruthenium
red. It is described in some of the older literature by Luft, I
think.
Carol

} Date: Mon, 14 Nov 2005 09:25:52 -0600
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--
__
Carol A. Heckman, Ph.D.
Professor of Biological Sciences
Director, Center for Microscopy & Microanalysis
Bowling Green State University, Bowling Green, OH 43403
fax: (419) 372-2024 email: heckman-at-bgnet.bgsu.edu
http://www.bgsu.edu/departments/biology/facilities/MnM
___________________________________________________________________________

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From: DFORAN-at-ORA.FDA.GOV
Date: Mon, 14 Nov 2005 10:20:11 -0600
Subject: [Microscopy] FW: Re: AskAMicroscopist: 10 year old son who wants

Contents Retrieved from Microscopy Listserver Archives
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Try your local microscope repair service. Often the have refurbished good
to high quality used microscopes.

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On Nov 10, 2005, at 3:40 PM, foxglovelj-at-msn.com wrote:

} Email: foxglovelj-at-msn.com
} Name: Laura Giard
}
} Education: Undergraduate College
}
} Location: Sterling, MA USA
}
} Title: microscope purchase
}
} Question: Hello, I have a 10 year old son who wants a microscope for
} xmas. He wants to be a scientist when he grow up! We have purchased
} "cheap" microscopes over the years and have been very disappointed.
} I'd like to find one that truly works, however, cannot spend a
} fortune. Can you give me some help? Magnifications etc. do not
} really help me in determining the quality which is the only
} information I can seem to gain from places that sell them. Right now
} I've been looking at ones through Nasco. ANY HELP would be GREATLY
} APPRECIATED! Thank you.
}
Dear Laura,
Check out Project MICRO; go to the MSA web site
(www.msa.microscopy.org), click on the Reference & Educational button
on the left side, and then on ProjectMICRO, the 6th item under
Education Committee Functions.
Yours,
Bill Tivol, PhD
EM Scientist and Manager
Cryo-Electron Microscopy Facility
Broad Center, Mail Code 114-96
California Institute of Technology
Pasadena CA 91125
(626) 395-8833
tivol-at-caltech.edu


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From: gwe-at-ufl.edu
Date: Mon, 14 Nov 2005 10:46:08 -0600
Subject: [Microscopy] Re: SEM imaging of bacteria encapsulation

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

Freeze drying might be the best bet for holding on the everything
without adding artefactual material such as Ru.

hagglundk1-at-nku.edu wrote:

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From: hyi-at-emory.edu
Date: Mon, 14 Nov 2005 11:56:33 -0600
Subject: [Microscopy] CCD camera for TEM

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

Good afternoon, Everyone:
   
We are in the process to get a CCD camera for our TEM. I am not very
familiar with the market. Does anyone have any suggestion in terms of
which companies are reliable and offer good customer service all
that. I remember there was a user gathering a few years ago at MSA
meeting to discuss problems with one particular company. I was not at
that gathering so did not know a lot of details.  Can anyone tell me
what are the issues when buying a CCD camera? 

            Hope to hear your opinions. Thank you in advance.


Hong
Emory EM



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From: tivol-at-caltech.edu
Date: Mon, 14 Nov 2005 12:41:55 -0600
Subject: [Microscopy] Re: SEM imaging of bacteria encapsulation

Contents Retrieved from Microscopy Listserver Archives
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On Nov 14, 2005, at 7:24 AM, hagglundk1-at-nku.edu wrote:

} We have a student working on a project in which they are growing
} bacterial cultures (pseudomonas and other species) in a hostile
} environment. The bacteria form capsules that we are interested in
} seeing. The capsules are most likely composed of sugars
} (polysaccharides), and we are hoping to find a fixation or other
} preparation technique that would help us with imaging these. We have
} so
} far prepared cultures with standard fixation, dehydration, and critical
} point drying techniques, but are not sure that we are preserving the
} capsules forming around the bacteria.
}
} We do not have a cryo stage available.
}
Dear Karl,
Two possibilities are negative stain and cryofixation. Even though
you do not have a cryostage, high-pressure freezing followed by
freeze-substitution and embedding will give you the specimen in a block
of resin that can be stained and sectioned in the conventional manner.
Of course, you may not have access to a high-pressure freezer.
Yours,
Bill Tivol, PhD
EM Scientist and Manager
Cryo-Electron Microscopy Facility
Broad Center, Mail Code 114-96
California Institute of Technology
Pasadena CA 91125
(626) 395-8833
tivol-at-caltech.edu


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From: PWebster-at-hei.org
Date: Mon, 14 Nov 2005 12:58:27 -0600
Subject: [Microscopy] Re: SEM imaging of bacteria encapsulation

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

Dear Karl,

You might like to try a simplified cryo-fixation method. It is not the best
method for many reasons and may not give you an impressive result as the
high-pressure freezer that Bill Tivol suggests, but it will give you an easy
way into these methods. Hopefully the results will give you enough data for
a grant!

First freeze your bacteria by immersion freezing in as close to the growing
state as possible. If they are growing in suspension, spin them down very
gently and scoop out the pellet to place onto a metal holder (any small
piece of wire or metal that can hold a drop of the suspension will work). If
they are growing on a substrate, leave them there.

Take the bacteria and freeze them by immersion in an efficient cryogen. We
use liquid propane from a bottle we get from a camping store (for cooking
food outdoors) - disclaimer here: propane is inflammable so take suitable
precautions not to blow up the lab. We collect the propane by piping it into
a small plastic cup that is surrounded by liquid nitrogen.

Once the bacteria have been frozen, which is almost immediately after they
have been immersed in the propane, transfer them to vials on dry ice,
containing dry ethanol or acetone. The dry ice is held in a styrofoam box
Leave them on the dry ice for a few days (depending on the size of the
specimen), changing the cold solvent with fresh cold solvent every day.

Transfer the specimens to the fridge (4 degrees) in a small styrofoam box
containing a small amount of dry ice and leave the specimens to warm to 4
degrees. The trick is to let the vials warm up slowly enough to not have the
solvent boil.

Once you have the solvent at 4 degrees, the specimens can be transferred to
a critical point drier and processed for SEM examination.

As I said, this is not the best way to prepare specimens using cryomethods
but you may be surprised by the results you get.

Paul Webster.




Paul Webster, Ph.D
House Ear Institute
2100 West Third Street
Los Angeles, CA 90057
(213) 273 8026
pwebster-at-hei.org





On 11/14/05 7:30 AM, "hagglundk1-at-nku.edu" {hagglundk1-at-nku.edu} wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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}
} We have a student working on a project in which they are growing
} bacterial cultures (pseudomonas and other species) in a hostile
} environment. The bacteria form capsules that we are interested in
} seeing. The capsules are most likely composed of sugars
} (polysaccharides), and we are hoping to find a fixation or other
} preparation technique that would help us with imaging these. We have so
} far prepared cultures with standard fixation, dehydration, and critical
} point drying techniques, but are not sure that we are preserving the
} capsules forming around the bacteria.
}
} We do not have a cryo stage available.
}
} Any ideas?
}
} Karl Hagglund
} Biological Sciences, SC-102B
} Northern Kentucky University, Nunn Drive
} Highland Heights, KY 41099
} 859-572-5238
} http://semlab.nku.edu
}
}
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From: gcc-at-couger.com
Date: Mon, 14 Nov 2005 13:56:14 -0600
Subject: [Microscopy] Re: FW: Re: AskAMicroscopist: 10 year old son who wants

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

} } Title: microscope purchase
} }
} } Question: Hello, I have a 10 year old son who wants a microscope for
} } xmas. He wants to be a scientist when he grow up! We have purchased
} } "cheap" microscopes over the years and have been very disappointed.
} } I'd like to find one that truly works, however, cannot spend a
} } fortune. Can you give me some help? Magnifications etc. do not
} } really help me in determining the quality which is the only
} } information I can seem to gain from places that sell them. Right now
} } I've been looking at ones through Nasco. ANY HELP would be GREATLY
} } APPRECIATED! Thank you.
} }
}
Dear Laura,

I have a page that I put together for the first time buyer of microscopes
http://www.couger.com/microscope/links/gcnewbuy.html

While I am normally very strongly in favor of buying older used
microscopes in the case of young children it is almost impossible to find
what they need.

If you buy used buy from a dealer if you buy on Ebay research the sellers
feedback and other sales very carefully. There are some good dealers on
ebay but you have to do you home work.

Gordon.


========= From My Page ===
Special Consideration for Children.


Microscopes for children present special considerations. First it is
important that the microscope be an instrument that they use and not play
with a few days and discard. In these days of the Internet and instant
gratification holding my interested is difficult holding a a child's is a
real challenge. Getting a scope too complex for them to use on their own
is a sure way to put most kids off. I know it did me over 50 years ago and
many complex procedures still do.

Another problem is young children's eyes are closer together than most
binocular tubes will close up and their binocular vision not as well
developed so binocular heads may pose a problem for them well into their
teens. Even if the scope will close down enough to accommodate their eyes
the muscular coordination of their eyes may not work with the binocular
head pieces. Having multiple sclerosis I have experienced this and it
quickly causes eyestrain if the eyes do not comfortably lock in on the image.

Of course having eyepieces and other parts that are attached so that they
are difficult to remove and lose or injure the child are important.

It is difficult to find this qualities in a used microscopes so you are
left with little choice but to buy from a dealer in new microscopes. I
hesitate to recommend particular dealer. I don't hesitate to strongly
recommend that you by from someone that has been a microscope dealer for
some years and has a good reputation for marinating the equipment they
sell. Buying from retail stores that have no microscope service department
is no different than buying from ebay except it is easy to find the person
to complain to when things don't work. It is not much easier to get them
fixed in most cases.

For their fist scope a low power 10 to 30 power scope that requires little
or no surface preparation may well hold their interest much longer than a
height power compound scope that require specimen preparation, careful
lighting and scope adjustment to get a decent image. All this is speaking
in generalities there are 6 year old kids that can master a complex scope
just as there are grown men than can't work a magnifying glass.

While I don't recommend any particular seller I use this scope as an
example of a good low power scope for the beginner
http://www.microscopeworld.com/low/lpdis.htm it is a 20x scope that has
all pieces attached so they aren't remove by curious hands and the
monocular set up allow easy use with out constant readjustment among users
and children that have problems with binocular vision have no problems.

Scopes like these http://www.microscopeworld.com/high/hpin.htm are very
good choice for youngsters. Get one with a simple condenser. I allows you
to work with them and the light and condenser actual adjustable stops are
simple enough children can use them unaided. Less expensive used scopes
can be purchased with better lenses but you will be hard pressed to find a
simpler scope than this one. Defused light and simple stop system is more
that adequate and much less frustrating than an adjustable condenser and
iris 40x and lower powered objectives. The contrast suffers a little at
40x but it is still good enough for very good viewing. Actual test
surprised me.

If the child has a real interest in science I would let him try some more
complex scopes and see what he can handle and try to come to a realistic
conclusion of his desires, abilities and drive to complete projects before
getting a complex microscope.



If you are considering a microscope for a gift to a child consider the
level of complexity the child can deal with. Condensers can give them a
hard time so get one with a simple condenser system or be prepared to
spend a lot of time with them if you get a complex one. Also binocular
heads are a problem. They may not close to the point that they can use
them and until their middle teens there binocular vision may not develop
to be able to use them very well.

In used compound scopes I have used the AO Spencer 160 with very good
results.

For the younger child I have found nothing better than the this Asian
import. http://www.microscopeworld.com/low/lpdis.htm

Occasionally a monocular dissection scope comes up on ebay but not often.
These scopes take little or no preparation of specials and are less likely
to end up in the closet if the child finds things to complex. For a
binocular scope AO Spencer Cycloptics sell for $ 50 and up on ebay and
www.roseoptics.com has some serviced and rebuilt ones that have the prisms
attached with modern cement that he sells for a reasonable price with a
guarantee. These old Cycloptics are ideal for children because they are so
sturdy and almost any one can clean sand dirt out of them if they
children get careless. and they offer 7 to 25x or 14 to 50x with a 2x lens.


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From: lkrupp-at-us.ibm.com
Date: Mon, 14 Nov 2005 14:11:38 -0600
Subject: [Microscopy] TEM sample storage

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Hello-

I was wondering how other listers store their TEM samples between
preparation and microscope time. We have oxidation problems, and I am
curious whether it is better to store samples under vacuum or an inert
gas, as far as preventing/delaying oxidation. Right now we store ours in
a cabinet pumped out by a diaphragm pump, and we see contamination even
after a couple days on our sensitive samples.

Thank you,
Leslie

Leslie Krupp (Thompson)
IBM Almaden Research
650 Harry Road, K19/D2
San Jose, CA 95120-6099

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From: m.obrien-at-sgul.ac.uk
Date: Mon, 14 Nov 2005 15:02:49 -0600
Subject: [Microscopy] viaWWW: help with Colorview12 camera and GrabBit card.

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Email: m.obrien-at-sgul.ac.uk
Name: Marc O'Brien

Organization: St George's University of London

Title-Subject: [Filtered] help with Colorview12 camera and GrabBit card.

Question: Hi

We have an Olympus BX51 with a Soft Imaging Systems
framegrabber camera and software, we've tried
upgrading the PC but have found that PCI GrabBit
card is not recognised by the motherboard. SIS say
they can upgrade the card for around 800 euros, but
can't supply a loan card while the GrabBit is away
being upgraded. They also state that the colorview
12 camera will not work with any other framegrabber
card .

We are loathe to dismantle our only working system.

Our only option would seem to be buying or borrowing
a spare PCI grabBit, and then having the upgrade
done, thus keeping a working system.

Does anyone have one of these cards spare?

Does anyone know of other framegrabber cards that
work with the colorview12 camera and Analysis 5
software?

Has anyone been through this process with SIS and
have any comments.

Any other thoughts or comments gratefully received.

Thanks

Marc O'Brien

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From: r.sims-at-auckland.ac.nz
Date: Mon, 14 Nov 2005 15:13:16 -0600
Subject: Re: [Microscopy] re: Dry pump emission

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Hi Steve

Yes, I'm sure that you are correct, but I STILL don't think that the term "carcinogenic"
should be used lightly, and without some foundation.

We're supposed to be scientists, aren't we?

We can't just call all nasties "carcinogenic" just because they're unhealthy..

cheers

rtch



Send reply to: "Steve Chapman" {protrain-at-emcourses.com}
X-from: "Steve Chapman" {protrain-at-emcourses.com}
To: {r.sims-at-auckland.ac.nz}



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From: papalia-at-udel.edu
Date: Mon, 14 Nov 2005 15:14:52 -0600
Subject: [Microscopy] Re: TEM sample storage

Contents Retrieved from Microscopy Listserver Archives
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Hi Leslie,

We've had some success in preserving easily oxidized samples by using
a rotary vane vacuum pump, grade 5 argon, and a vacuum
desiccator. We just put the samples in the desiccator (with or
without desiccant - your choice for your application), then triple
purge it with vacuum & argon, ultimately leaving it under active
vacuum. When ready for the samples, back-filling with the argon
again will buy you enough time to open up the desiccator and remove
your sample. We've seen a definite difference between the use of
nitrogen and the use of argon with argon being better at displacing
the oxygen in the system.

Hope that helps,
John

} Hello-
}
} I was wondering how other listers store their TEM samples between
} preparation and microscope time. We have oxidation problems, and I am
} curious whether it is better to store samples under vacuum or an inert
} gas, as far as preventing/delaying oxidation. Right now we store ours in
} a cabinet pumped out by a diaphragm pump, and we see contamination even
} after a couple days on our sensitive samples.
}
} Thank you,
} Leslie
}
} Leslie Krupp (Thompson)
} IBM Almaden Research
} 650 Harry Road, K19/D2
} San Jose, CA 95120-6099

John Papalia
Galvin Research Group
201 duPont Hall
Dept. of Materials Science & Engineering
University of Delaware
Newark, DE 19716



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From: schooley-at-mcn.org
Date: Mon, 14 Nov 2005 19:37:27 -0600
Subject: [Microscopy] Re: AskAMicroscopist: What kind of microscope should

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Per - You're being a bit too ambitious; start with larger organisms.
No one looks at living viruses; they're so small that an electron
microscope is needed. Dead & stained bacteria can be seen with a
compound microscope. If you want some fun & action, start with
protozoa in pond water. Look at the MICRO bibliography & order a
book or two. "Explore the world using protozoa" &/or "Guide to
microlife", and "Exploring with the microscope". Start with one of
the many Chinese scopes, perhaps with LED illumination. Get one that
you can add an 100x oil immersion objective to, AFTER you learn how
to use it. Then you'll be able to see bacteria, but they aren't
going to be very exciting to watch.
--
Caroline Schooley
Project MICRO Coordinator
Microscopy Society of America
Box 117, 45301 Caspar Point Road
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.microscopy.org/ProjectMICRO

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From: alex.titkov-at-millenniumchem.com
Date: Tue, 15 Nov 2005 00:55:20 -0600
Subject: [Microscopy] INCA Energy software

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

Hi,

Can anyone please tell me what the latest issue of The Microanalysis Suite
from Oxford Instruments is currently available?
What issue are you using? We are on issue 13. Our service contract entitles
us for free software upgrades, but we just can not get the information from
the local Oxford representatives.

Many thanks,
Alexander Titkov

Millennium Inorganic Chemicals Ltd
A Lyondell Company






Lyondell Chemical Company

The information contained in this e-mail message and any attachments may be confidential. It is intended only for the use of the individual or entities named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by e-mail at the originating address.

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From: ron.doole-at-materials.ox.ac.uk
Date: Tue, 15 Nov 2005 02:14:09 -0600
Subject: [Microscopy] INCA Energy software

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Hi Alex,

I am running Issue 15.

If you are having problems locally contact the Head Office in UK. They have
always been very helpful to me.

Regards,
Ron

-----Original Message-----
X-from: alex.titkov-at-millenniumchem.com [mailto:alex.titkov-at-millenniumchem.com]

Sent: 15 November 2005 07:04
To: ron.doole-at-materials.ox.ac.uk

Hi,

Can anyone please tell me what the latest issue of The Microanalysis Suite
from Oxford Instruments is currently available?
What issue are you using? We are on issue 13. Our service contract entitles
us for free software upgrades, but we just can not get the information from
the local Oxford representatives.

Many thanks,
Alexander Titkov

Millennium Inorganic Chemicals Ltd
A Lyondell Company






Lyondell Chemical Company

The information contained in this e-mail message and any attachments may be
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named above. If the reader of this message is not the intended recipient,
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this communication is strictly prohibited. If you have received this
communication in error, please notify us immediately by e-mail at the
originating address.

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From: gary-at-gaugler.com
Date: Tue, 15 Nov 2005 02:16:02 -0600
Subject: [Microscopy] Re: viaWWW: help with Colorview12 camera and

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I have a GrabBit PCI card you can borrow. $20 and a
promise to return it. You pay for shipping out and back.
$995 if you want to keep it.


gary g.



At 01:04 PM 11/14/2005, you wrote:



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From: kunli218-at-yahoo.com
Date: Tue, 15 Nov 2005 07:48:15 -0600
Subject: [Microscopy] viaWWW: Plan view TEM sample prep contamination

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Email: kunli218-at-yahoo.com
Name: Simon Lee

Organization: CharteredSEMi

Title-Subject: [Filtered] MListserver:Plan view TEM sample prep contamination

Question: Dear Listers,

We met some contamination problem when preparing plan-view TEM sample where the sample can only be milled from one side. We found that the contamination at the unmilled side is quite significant (redeposition). The miller we use is PIPS from Gatan. Will the graphite post help? and Is there any other procedure to minimize the contamination?

Thanks in advance for your help.

Rdgs,

Simon

Chartered Semiconductor

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

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From: U.J.Potter-at-bath.ac.uk
Date: Tue, 15 Nov 2005 07:55:05 -0600
Subject: [Microscopy] viaWWW: Resin/wax sectioning problems

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Email: U.J.Potter-at-bath.ac.uk
Name: Ursula Potter

Organization: The University of Bath UK

Title-Subject: [Filtered] MListserver: Resin/wax sectioning problems

Question: Dear All,

We have a problem with the sectioning of Zebrafish embryos (early stages with yolk sac). The embryos have been subjected to an in situ hybridization staining method and when subsequently embedded in wax or resin (technovit) the sections show very bad chatter (wax) or tearing (resin) at the area of the yolk sac. Has anyone experienced this problem? Advice to solve problem would be gratefully received.

Thanks
Ursula

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From: eschumacher-at-mccrone.com
Date: Tue, 15 Nov 2005 08:03:20 -0600
Subject: [Microscopy] INCA Energy software

Contents Retrieved from Microscopy Listserver Archives
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Hi Alexander,

I just received Issue 16 last week as a service contract upgrade. We've
always gotten prompt responses on software and other issues from our
local service and sales reps and from the MA office, with additional
support from the UK if necessary. Getting the upgrade with your service
contract renewal should be straightforward.

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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-----Original Message-----
X-from: alex.titkov-at-millenniumchem.com
[mailto:alex.titkov-at-millenniumchem.com]
Sent: Tuesday, November 15, 2005 12:56 AM
To: Elaine F. Schumacher

Hi,

Can anyone please tell me what the latest issue of The Microanalysis
Suite
from Oxford Instruments is currently available?
What issue are you using? We are on issue 13. Our service contract
entitles
us for free software upgrades, but we just can not get the information
from
the local Oxford representatives.

Many thanks,
Alexander Titkov

Millennium Inorganic Chemicals Ltd
A Lyondell Company






Lyondell Chemical Company

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From: hagglundk1-at-nku.edu
Date: Tue, 15 Nov 2005 09:02:32 -0600
Subject: [Microscopy] Imaging bacteria capsules Thanks

Contents Retrieved from Microscopy Listserver Archives
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Thanks to everyone who responded to my question about imaging bacteria.
We have a freeze dryer available, and will likely attempt using this
first. The best part is that our student has plenty of directions to
continue her research.

Karl Hagglund
Biological Sciences, SC-102B
Northern Kentucky University, Nunn Drive
Highland Heights, KY 41099
859-572-5238
http://semlab.nku.edu


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From: richard.beanland-at-bookham.com
Date: Tue, 15 Nov 2005 09:18:31 -0600
Subject: [Microscopy] viaWWW: Plan view TEM sample prep contamination

Contents Retrieved from Microscopy Listserver Archives
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Hi Simon,
I have a PIPS which I use for cross sections and the occasional plan view. I find it is not too bad, but I usually have to give the original surface a tickle with the ion beam to clean off contamination. I generally use the standard double-sided holder, not the graphite post (since I have never been very confident about getting the sample off the post again without breaking it!) Milling conditions are double modulation, 3 degrees incidence from below with the top surface up (i.e. facing the viewing port), slowest possible rotation, 6kV. When the sample is thin enough I turn the voltage down to 2.5 kV (the gas flow has to be adjusted to get maximum current) to give a final clean of the milled surface for a minute or two. The top surface clean is also at 2.5 kV but only one burst of the gun, probably about 12 seconds. This has worked quite well, I have identified nm scale contamination on SiO2 films without any real problems with artefacts. It probably helps to stop milling as soon as you get the smallest hole.
If I really have to keep the top surface intact I prefer using chemical methods, jet etching with Cl in methanol for III-Vs or HF:HNO3 for Si.

Good luck!

Richard

________________________________________
Richard Beanland
Analytical Services
Bookham Inc
Caswell
Towcester
Northants
NN12 8EQ
United Kingdom
Tel. +44 1327 356362
Fax. +44 1327 356775
http://www.bookham.com
________________________________________


-----Original Message-----
X-from: kunli218-at-yahoo.com [mailto:kunli218-at-yahoo.com]
Sent: 15 November 2005 13:50
To: Richard Beanland

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Email: kunli218-at-yahoo.com
Name: Simon Lee

Organization: CharteredSEMi

Title-Subject: [Filtered] MListserver:Plan view TEM sample prep contamination

Question: Dear Listers,

We met some contamination problem when preparing plan-view TEM sample where the sample can only be milled from one side. We found that the contamination at the unmilled side is quite significant (redeposition). The miller we use is PIPS from Gatan. Will the graphite post help? and Is there any other procedure to minimize the contamination?

Thanks in advance for your help.

Rdgs,

Simon

Chartered Semiconductor

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

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From: lkrupp-at-us.ibm.com
Date: Tue, 15 Nov 2005 09:39:35 -0600
Subject: [Microscopy] Re: viaWWW: Plan view TEM sample prep contamination

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

Hi Simon-

For plan view samples I prefer to use the regular post sample holder. I
attach the sample with a tiny drop of crystal bond (I mean tiny!), which
then gets cleaned off manually with a pointed q-tip and acetone. I do not
like to soak off the samples because I feel it contaminates the area you
just milled. Also, stop milling when you get the smallest possible hole
or it will redep through the hole, and reduce the kV as you get closer to
finishing.

If crystal bond residue cannot be tolerated (of course you can't clean it
all off), then I use the graphite holder, with a 3mm disc cut from a glass
coverslip underneath the sample to protect the surface. The only problem
with the graphite holder is the slider bars on either side will block the
beam when using rotation only, and the hole will end up somewhat oblong.

If none of that works, you can mill the sample for a few seconds on the
'good' side, but this is hit or miss.

Sincerely,
Leslie

Leslie Krupp (Thompson)
IBM Almaden Research
650 Harry Road, K19/D2
San Jose, CA 95120-6099







Email: kunli218-at-yahoo.com
Name: Simon Lee

Organization: CharteredSEMi

Title-Subject: [Filtered] MListserver:Plan view TEM sample prep
contamination

Question: Dear Listers,

We met some contamination problem when preparing plan-view TEM sample
where the sample can only be milled from one side. We found that the
contamination at the unmilled side is quite significant (redeposition).
The miller we use is PIPS from Gatan. Will the graphite post help? and Is
there any other procedure to minimize the contamination?

Thanks in advance for your help.

Rdgs,

Simon

Chartered Semiconductor

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

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From: john.mardinly-at-intel.com
Date: Tue, 15 Nov 2005 10:40:25 -0600
Subject: [Microscopy] TEM sample storage

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

We use the Gatan 655 dry pump station with the TEM specimen holder
accessory. It is a bit pricy, but does an outstanding job, and it is
very quick and easy to use, with cycle time measured in seconds!
http://www.gatan.com/pdf/655%20Dry%20Pumping%20Station.pdf


John Mardinly
Intel Corporation

The opinions of this writer do not necessarily represent the opinion of
Intel Corporation.


-----Original Message-----
X-from: lkrupp-at-us.ibm.com [mailto:lkrupp-at-us.ibm.com]
Sent: Monday, November 14, 2005 12:12 PM
To: Mardinly, John

Hello-

I was wondering how other listers store their TEM samples between
preparation and microscope time. We have oxidation problems, and I am
curious whether it is better to store samples under vacuum or an inert
gas, as far as preventing/delaying oxidation. Right now we store ours
in
a cabinet pumped out by a diaphragm pump, and we see contamination even
after a couple days on our sensitive samples.

Thank you,
Leslie

Leslie Krupp (Thompson)
IBM Almaden Research
650 Harry Road, K19/D2
San Jose, CA 95120-6099

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From: bozhilov-at-ucr.edu
Date: Tue, 15 Nov 2005 10:41:02 -0600
Subject: [Microscopy] EDAX Genesis 3.6 Net intensity

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

Hi all,

I need to verify what do the Net Intensity numbers in the
quantification panel represent in the MThin version of the EDAX
Genesis 3.6 software.

According to the manuals: "Net Intensity is the Intensity of the peak
minus the background after de-convolution".

The problem I have is that using the value of the measured net
intensity listed after quantification, I cannot obtain the same
number for the Intensity error as listed by the software. I am using
the formula:
square root of (net intensity multiplied by the live seconds) divided
by the
(net intensity multiplied by the live seconds).

If I subtract the value of the intensity of the background from the
Net Intensity value then I get exactly the number listed as intensity
error in the software.

The question is now. Which is correct? The statement that the Net
intensity are peak intensity minus the background or that the Net
Intensity is the peak intensity + the background. Or may be I am not
using the correct calculation procedure for the intensity error?

I have sent this question to EDAX representatives about three weeks
ago and I am still waiting for an answer so I decide to "pool the
audience" as Regis Philbin likes put it.

_______________________________________
Krassimir N. Bozhilov
Central Facility for Advanced Microscopy and Microanalysis
University of California
Riverside, CA 92521

tel 951 927 2998
fax 951 827 2489
bozhilov-at-ucr.edu
_______________________________________



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From: walck-at-southbaytech.com
Date: Tue, 15 Nov 2005 12:11:01 -0600
Subject: [Microscopy] viaWWW: Plan view TEM sample prep contamination

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

There are a couple of ways to prevent contamination on the back side
during ion milling of a plan view sample. The most common way is to use
a lacquer that your remove afterwards. The one that I would recommend
is MicroShield or Microstop -I can't remember the name for sure and it
is available from SPI as well as the remover. Other people have used
nail polish. May I recommend Sally Hanson's Hard As Nails?

I can't find the reference for the neatest way that I think is the way
to do this. I thought that it was in the first MRS TEM sample Prep book
(Vol 115), but I couldn't find it there. What you do is evaporate NaCl
in a vacuum evaporator onto the side that you want to protect. Ion mill
on the other side. The NaCl layer is the one that is contaminated and
after perforation in the ion mill, you simply dip the sample in
distilled water to dissolve the NaCl and float off the contamination
layer. I wish that I could remember who did that, but I thought that it
was a pretty slick idea. If anyone knows the reference or who did it,
please let me know because I would like to put that in our application
notes section of our web site.


-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
walck-at-southbaytech.com


-----Original Message-----
X-from: kunli218-at-yahoo.com [mailto:kunli218-at-yahoo.com]
Sent: Tuesday, November 15, 2005 5:53 AM
To: Walck-at-SouthBayTech.com

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 kunli218-at-yahoo.com as well as the MIcroscopy
Listserver
------------------------------------------------------------------------
---

Email: kunli218-at-yahoo.com
Name: Simon Lee

Organization: CharteredSEMi

Title-Subject: [Filtered] MListserver:Plan view TEM sample prep
contamination

Question: Dear Listers,

We met some contamination problem when preparing plan-view TEM sample
where the sample can only be milled from one side. We found that the
contamination at the unmilled side is quite significant (redeposition).
The miller we use is PIPS from Gatan. Will the graphite post help? and
Is there any other procedure to minimize the contamination?

Thanks in advance for your help.

Rdgs,

Simon

Chartered Semiconductor

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

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From: frank.karl-at-degussa.com
Date: Tue, 15 Nov 2005 13:04:53 -0600
Subject: [Microscopy] in search of training....

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

I just had a EDS system installed on my TEM. As I'm still learning how to
use the TEM side of this equation, does anyone have any suggestions on
short courses or training I can get so I know what I'm doing?

Thanks in advance!!!

Frank Karl
Degussa Corporation
Akron Technical Center
3500 Embassy Parkway
Suite 100
Akron, Ohio 44333


330-668-2235 Ext. 238


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From: mgb-at-ansto.gov.au
Date: Tue, 15 Nov 2005 16:36:33 -0600
Subject: [Microscopy] GIF collection semi-angle

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

Hi All,

A colleague asked me to post the following question. We would
appreciate any feedback either directly to me or to the Listserver.
Cheers,

"Does any one know the value for GIF collection semiangle for a JEOL
2010F /GIF 2000 (200kV) when operated with an image on the screen, no
objective aperture present and a 3mm GIF entrance aperture?

If you have a value, I'd also appreciate the physical dimensions,
assumptions and calculations you have made in arriving at it?

Many thanks in advance."


Mark Blackford
Institute of Materials and Engineering Science, ANSTO
PMB 1,
Menai, N.S.W., 2234
Australia

Phone 61 2 9717 3027
Fax 61 2 9543 7179

Disclaimer:
The views expressed in this E-mail message do not necessarily
represent the official views of ANSTO from which this message was
conveyed.




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From: walck-at-southbaytech.com
Date: Tue, 15 Nov 2005 17:05:47 -0600
Subject: [Microscopy] TEM sample storage

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

I am not sure how to write this so that it does not sound like too much
of a commercial. I will put the disclaimer up front so that you can
choose to read on and I will try to be as factual about the product as I
can without too much commercialism and hope that I don't get spanked by
Nestor. I think that I am within the guidelines because what I am
relating below is the design considerations and features that evolved
out of solving the solution to my microscopy studies that I have not
reported on before.

Disclaimer: South Bay Technology manufactures and sells the
SampleSaver(TM) Portable Storage Container that addresses the problem of
transportation and storage of samples or samples with coatings that
react when exposed to the atmosphere. This product was just introduced
at the M&M 2005 meeting and other than that and the recent ISFTA 2005
meeting, we have not advertised it. In response to this question, we
just put the brochure for the unit on our web site in order so that
anyone interested can view what the unit looks like. This unit was
specifically made to address the general topic of this question.

The SampleSaver(TM) portable storage container (SS) is my first
contribution to the SBT product line and came about because of problems
that I had with reactive samples when I worked at Wright Patterson Air
Force Base and at PPG Industries, Inc. It is a plastic container in
which there are two valves, one of which is a conventional valve and one
which is integral into the body of the unit, itself. Metallurgical,
SEM, and TEM samples can be held inside with different "sample trees".
There is also a special unit that we have that is made for FIB lift-out
samples. The SS is designed to be purged without pumping it down with
an inert gas that could be N2, Ar, or CO2 from a cylinder or from a
special unit, which we also sell, will use the boil-off from liquid
nitrogen. The idea of using the boil-off from liquid nitrogen (or the
CO2 gas from Dry Ice) is that if you prepare your samples elsewhere and
travel to another lab to do the analysis or vice versa (e.g. when you
FIB your samples at another lab and bring them home) the other lab will
always have liquid nitrogen available to use as the source for the inert
gas. They may or may not have a vacuum pump, Ar line, or a N2 line
available for use. One major benefit of using LN2, is that the boil-off
from LN2 is extremely pure. When the SS unit is sufficiently purged,
the two valves are closed, body valve first then the gas supply valve.
The valve body can then be compressed. This then pressurizes the
container to help prevent ingress of any gas species by diffusion
through the plastic unit. The unit can be tested for integrity just
before opening by opening the valve while it is compressed. A "psst"
sound is reassuring. A tube from the valve can be put under LN2 so that
the container is not exposed to air.

I have images of XTEM samples of Low-E coatings on glass that have been
preserved using this system that I could share with anyone that is
interested. These Low-E coatings contain two layers of silver that when
the two surfaces of the cross section are exposed degrade in air over a
short period of time. Samples that have been stored for many weeks have
not undergone any degradation.

The unit was also developed for SEM samples that would be coated with
chromium where the samples would otherwise oxidize in a relative short
period. It was also intended for EBSD applications of metallurgical
materials where surface oxidation after preparation would degrade the
EBSD results.

Although I designed this only to be used in a purging mode because a
vacuum pump would be too bulky for portability and you can't guarantee
that one is available at the other lab, it can also be used with a
vacuum pump, if desired, and a pump-backfill sequence is possible with
it. I don't recommend leaving it in a vacuum condition, because of the
possible diffusion of species such as O2 and H2 through the plastic.
The positive pressure within the SS with an inert gas inhibits this
diffusion.

The SS can also be used with glove boxes such as the type that was
introduced by Omniprobe at M&M 2005. Here, the unit is simply put
inside the glove box and when the glove box is sufficiently purged,
samples can be put into it or removed from it and then reclosed. By
compressing the body, the pressure inside the SS will be higher than the
pressure in the glove box.

What we don't know at this time is whether samples stored in it and used
with it will require plasma cleaning for high resolution work. My
results have been good and I have not seen any contamination when used
with a LaB6 microscope. I have transported samples to field emission
microscopes and have not seen contamination issues. I suspect that the
higher pressure of the inert gas inside the SS may prevent or inhibit
any out gassing from the plastic body, but I am only guessing here.

-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
walck-at-southbaytech.com


-----Original Message-----
X-from: lkrupp-at-us.ibm.com [mailto:lkrupp-at-us.ibm.com]
Sent: Monday, November 14, 2005 12:15 PM
To: Walck-at-SouthBayTech.com

Hello-

I was wondering how other listers store their TEM samples between
preparation and microscope time. We have oxidation problems, and I am
curious whether it is better to store samples under vacuum or an inert
gas, as far as preventing/delaying oxidation. Right now we store ours
in
a cabinet pumped out by a diaphragm pump, and we see contamination even
after a couple days on our sensitive samples.

Thank you,
Leslie

Leslie Krupp (Thompson)
IBM Almaden Research
650 Harry Road, K19/D2
San Jose, CA 95120-6099

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From: rcommon-at-msu.edu
Date: Tue, 15 Nov 2005 18:01:37 -0600
Subject: [Microscopy] Fading Sections

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

After discovering that many of our older toluidine blue stained semi-thin
sections that had been mounted with a commercial mounting medium had faded,
I decided to try using our standard Epon-Araldite-DDSA embedding resin as a
mounting medium. Simply put a drop or two of resin (we use left over resin
from embeddings, stored in vials in a freezer) on the dry, stained slides,
and coverslip slowly to avoid bubbles. The slides are viewable immediately,
and will harden in a few days on their own, or overnight at 60 degrees.
They should not be overheated or some destaining or wrinkling may occur.
Since switching to Epon as a mounting medium, I have not seen any problem
with fading.

As for viewing slides before permanent mounting, the image can be greatly
improved just by placing a coverslip over the dry section while viewing.
Most objectives are corrected for the presence of a coverslip, so even with
nothing but air between the coverslip and the section, the image is very
good.

Ralph Common
Division of Human Pathology
Michigan State University


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From: alex.titkov-at-millenniumchem.com
Date: Tue, 15 Nov 2005 21:34:09 -0600
Subject: [Microscopy] INCA Energy software

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


Thanks everyone. I shall chase issue 16 then. Forgot to mention that we are
in Australia, not in the States as some people thought.

Cheers,
Alex

==================
Alexander Titkov
Millennium Inorganic Chemicals Ltd
A Lyondell Company
Locked Bag 245 Bunbury WA 6230
AUSTRALIA
Ph 08 9780 8505
FAX 08 9780 8500
E-mail: alex.titkov-at-millenniumchem.com




eschumacher-at-mccro
ne.com To: alex.titkov-at-millenniumchem.com
cc:
15/11/2005 10:06 Subject: [Microscopy] RE: INCA Energy software
PM
Please respond to
eschumacher









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Hi Alexander,

I just received Issue 16 last week as a service contract upgrade. We've
always gotten prompt responses on software and other issues from our
local service and sales reps and from the MA office, with additional
support from the UK if necessary. Getting the upgrade with your service
contract renewal should be straightforward.

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
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-----Original Message-----
X-from: alex.titkov-at-millenniumchem.com
[mailto:alex.titkov-at-millenniumchem.com]
Sent: Tuesday, November 15, 2005 12:56 AM
To: Elaine F. Schumacher

Hi,

Can anyone please tell me what the latest issue of The Microanalysis
Suite
from Oxford Instruments is currently available?
What issue are you using? We are on issue 13. Our service contract
entitles
us for free software upgrades, but we just can not get the information
from
the local Oxford representatives.

Many thanks,
Alexander Titkov

Millennium Inorganic Chemicals Ltd
A Lyondell Company






Lyondell Chemical Company

The information contained in this e-mail message and any attachments may
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Lyondell Chemical Company

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From: monica.iliescu-at-polymtl.ca
Date: Wed, 16 Nov 2005 06:59:12 -0600
Subject: [Microscopy] viaWWW: ESEM meetings?

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Email: monica.iliescu-at-polymtl.ca
Name: Monica ILIESCU

Organization: Ecole Polytechnique

Title-Subject: [Filtered] MListserver:

Question: Dear colleagues,
I would like to ask if someone of you know conferences involving (environmental) electron microscopy and biomaterials or related biological materials taking place in 2006.
Thank you very much.

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6, 12 -- Subject: viaWWW: ESEM meetings?
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From: MSHERWOOD-at-PARTNERS.ORG
Date: Wed, 16 Nov 2005 08:23:45 -0600
Subject: [Microscopy] re: New England Society for Microscopy (NESM) Fall Symposium

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

The Fall Symposium of the New England Society for Microscopy will once again be
held at Gordon College in Wenham, MA. It will be held on Thursday, December 1st
from Noon - 8:30pm. The program highlights talks on various microscopy
techniques. For more detailed information, including registration, the speakers
and talks, please go to NESM's website: http://nesm.cims.harvard.edu and click
on "current newsletter".

The deadline for registration (including dinner) is Monday, November 28th.
Please contact Paul Bain, Treasurer at Paul_Bain-at-hms.harvard.edu.

We look forward to seeing you at the meeting!

Peggy Sherwood
Corresponding Secretary, NESM







Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (W224)
Massachusetts General Hospital
55 Fruit Street
Boston, MA 02114
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-3192 (fax)
msherwood-at-partners.org


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From: klk-at-biotech.ufl.edu
Date: Wed, 16 Nov 2005 14:23:28 -0600
Subject: [Microscopy] starch in sections

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

I am looking for a technique that will help identify starch in
osmicated/Embed-Araldite embedded sections. One suggestion has been to
use amylase digestion or amylase/gold. Any advice or other suggestions
would be greatly appreciated.

--
Karen L. Kelley
ICBR Electron Microscopy Manager
University of Florida
ICBR Electron Microscopy Core Lab
Bartram Hall Room 214
Box 118525 Gainesville Florida
Lab: 352-392-1184 fax: 352-846-0251
Email: klk-at-biotech.ufl.edu
Southeastern Microscopy Society Treasurer
http://www.biotech.ufl.edu/EM/



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From: baskin-at-bio.umass.edu
Date: Wed, 16 Nov 2005 16:56:18 -0600
Subject: [Microscopy] Re: starch in sections

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

Hi Karen,
Starch grains are beautifully birefringent so they show up a
treat in polarized light microscopy. If you can use semi thin
sections to find/demonstrate them, you would be home free. In an
ultra-thin section, the polarized light signal would be low, so you
might need a sensitive instrument to detect them but I think still
quite possible. Of course this is at the light level, not TEM. Hope
it helps.

Tobias Baskin

}
}
} I am looking for a technique that will help identify starch in
} osmicated/Embed-Araldite embedded sections. One suggestion has been to
} use amylase digestion or amylase/gold. Any advice or other suggestions
} would be greatly appreciated.
}
} --
} Karen L. Kelley
} ICBR Electron Microscopy Manager
} University of Florida
} ICBR Electron Microscopy Core Lab
} Bartram Hall Room 214
} Box 118525 Gainesville Florida
} Lab: 352-392-1184 fax: 352-846-0251
} Email: klk-at-biotech.ufl.edu
} Southeastern Microscopy Society Treasurer
} http://www.biotech.ufl.edu/EM/
}
}
}
} =======


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

==============================Original Headers==============================
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From: barbara.miner-at-intel.com
Date: Wed, 16 Nov 2005 21:41:34 -0600
Subject: [Microscopy] viaWWW: Job Opening at INTEL , TEM sample prep technician

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Email: barbara.miner-at-intel.com
Name: Miner, Barbara

Organization: INTEL

Title-Subject: [Filtered] Experienced TEM sample prep technician

Question: Job Title: experienced TEM sample prep technician
Location: Hillsboro, OR
Start date: December 2005 or January 2006
Requisition will close at the end of November

Intel Corporationís Materials Analysis Lab, supporting development of next generation micro-processors, has immediate openings for experienced Transmission Electron Microscopy sample prep technicians in Hillsboro, Oregon. ÝÝSuccessful applicants must have experience working with patterned Si wafers, operating dual-beam FIBs, and applying precision mechanical polish. Candidate must have excellent attention to detail and the ability to multi-task. Good communication, problem-solving skills, and cooperative team work are mandatory. The work shift includes one weekend day. Relocation assistance is provided.

As the world's largest chip manufacturer, Intel strives to make every facet of semiconductor manufacturing state-of-the-art -- from semiconductor process development and manufacturing, through yield improvement to final test and optimization, and lastly packaging. Employees in the Technology and Manufacturing group are part of a worldwide network of manufacturing and assembly/test facilities.

Intel only accepts resumes/CV's submitted to our web site. To enable us to process your application, please submit your resume/CV to www.intel.com/jobs. ÝPlease, also send mail to {mailto:Barbara.miner-at-intel.com} Barbara.miner-at-intel.com indicating your intent to apply. ÝPlease note the job title and location to complete your search and application. Intel hires qualified candidates who are authorized to work in the U.S.-- that is, authorized to work without restriction as to a particular employer. This includes U.S. citizens or nationals, U.S. legal permanent residents, temporary residents granted legalization under the Immigration Reform and Control Act of 1986, asylees, and refugees. For foreign nationals who do not fall in one of the above categories, we limit our hiring of persons requiring visa sponsorship or individuals currently on a non-immigrant visa (e.g., H-1, J-1, L-1, F-1, B-1, TN) to candidates at the MS and PhD levels (or those who have equivalent work experience) who are applying for positions for which there is a demonstrated shortage of qualified U.S. candidates. At Intel, we are committed to equal employment opportunity. We respect, value and welcome diversity in our workforce, as well as in our customers, our suppliers and the global marketplace. Intel also values being a great place to work and strives to maintain a safe and drug-free workplace. Accordingly, Intel conditions all offers of employment on satisfactory completion of a drug screen (where allowed) and a background check. Intel does not accept resumes from headhunters or suppliers that have not signed a formal fee agreement. Our supplier base is limited to specific hiring needs. Therefore, any resume received from an unapproved supplier will be considered unsolicited, and Intel will not be obligated to pay a referral fee.


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From: mike.reedy-at-cellbio.duke.edu
Date: Thu, 17 Nov 2005 01:27:41 -0600
Subject: [Microscopy] Re: SEM imaging of bacteria encapsulation

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

Karl,
Just to offer a heretically simplifed option for quick-and-dirty (but
often unexpectedly clean) speedy results:
I agree with Muss that tannic acid is great, the surest simple
fixative, and with Webster that freezing is a promising approach.

But I suggest that for fixing extracellular matrix, and intracellular
structures of detergent (TX-100) pre-permeabilized cells, you can do
quite well without OsO4; just use freshly dissolved 0.2% TA in your
physiol extracellular or intracellular buffer at pH 6.8-7.0 (absent
any components that precipitate in TA) for 30 min, then wash out all
unbound TA in 5 buffer changes, and follow with 1% UrAc in DI water,
no pH adjustment needed, then do acetone or EtOH dehydration as fast
as you like. If you wish, you can interpose 0.5-2.0 hr in 1%
glutaraldehyde after the TA and before the UrAc, but i usually use
straight TAURAC as a preferred binary fix; is certainly less toxic
and just as fixing as a similar TAOS procedure.
(M. K. Reedy, C. Lucaveche, D. Popp, Biophysical Journal 59,
579a (1991)).
Worth one shot for comparison is using a TA-glut mixture as a trial
primary fix before UrAc, but I find glut sometimes tends to rush in
and perturb orderly lattice arrays that TA alone fixes more slowly
and beautifully. TA primary fix MUST be stabilized by UrAc or OsO4
secondary fix before dehydration.

I've used the same TAURAC sequence in cryo-acetone for
freeze-substitution almost exclusively for 15 years. that's what
suggests to me its possible value for polysaccharides. It is able to
fix and stabilize (or perhaps it just permits PHYSICAL in-situ
stabilizing of) the threadlike molecules of 500,000 MW dextran we
sometimes use at 3-5% as an osmotic squeezer for the myofilament
lattice of glycerinated muscle fibers. We can see them in thin
sections, excluded to the space between myofibrils. However, the
aqueous TAURAC fix does NOT preserve /retain the free dextran
molecules, so I guess the quick-freezing and/or acetone is needed for
that.

So I guess maybe using an initial non-cryo "fix" of acetone alone,
followed by TAURAC in acetone and comparing with acetone-TAURAC
sequence and with an acetone-URAC-TA sequence might be worth trying.
Note Craig lab's findings that UrAc alone can be a fabulous fixative.
(F. Q. Zhao, R. Craig, Journal of Structural Biology 141, 43
(Jan, 2003).

And if you want to try a totally safer alternative than propane for
the plunge-freezing that Paul Webster suggests, LN2 plunging has a
bad rap that is undeserved. If you plunge rapidly through 10-12
inches of LN2, you leave behind all the insulating bubbles
responsible for the Leidenfrost effect and get convective cooling at
least as rapid or more-so than propane provides. See what protein
crystallographers from Hope's lab have to say about how reliable this
can be!
(L. J. Walker, P. O. Moreno, H. Hope, Journal of Applied
Crystallography 31, 954 (1998); S. Parkin, H. Hope, Journal of
Applied Crystallography 31, 945 (1998)).

Best of luck!
--
-mike reedy-

************************
Michael K. Reedy, M.D.
Duke Univ. Med. Center
Dept. Cell Biology, Box 3011 (for U.S. Mail)
458 Alex Sands Bldg, Research Dr (courier)
Durham, NC 27710

Office 919-668-2534
Lab 919-684-5674
Fax 919-681-9929
mike.reedy-at-cellbio.duke.edu
http://note.cellbio.duke.edu/Faculty/Research/Reedy.html

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From: justgetmeonhere-at-netscape.net
Date: Thu, 17 Nov 2005 07:13:46 -0600
Subject: [Microscopy] viaWWW: question about BSE detectors

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Email: justgetmeonhere-at-netscape.net
Name: Matthew McCaskill

Organization: none

Title-Subject: [Filtered] question about BSE detectors

Question: Hello all,

I am enrolled in an SEM class, and for this class I have been asked to do a paper on backscattered electron (BSE) detectors. Specifically, I have to briefly state what is state-of-the-art now, and then discuss research or improvements that are occurring right now. Then I am to predict and forecast the future of the technology; will there be more developments, or have BSE detectors fully matured technologically?

The point of all this post, however, is that I'm having trouble finding literature on SEM in general, let alone BSE detectors. Utilizing my school's library and the internet, I have found at least four articles already, but I need at least four more. Can anybody recommend me some journal articles or papers that deal with my topic? Preferably, I'd like for them to be available as FREE electronic copies (I found a few promising articles online, but unfortunately I have to pay to view, which is disadvantageous to poor college students such as myself), but if I can get ahold of the actual hard copies at my school's library then that should also be enough.

Thanks to all who can help me in this regard, and thanks to all of you for your time.

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From: L.Tilley-at-latrobe.edu.au
Date: Thu, 17 Nov 2005 07:21:37 -0600
Subject: [Microscopy] viaWWW: Post Doc Position Open - ultrastructural biology

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Email: L.Tilley-at-latrobe.edu.au
Name: Leann Tilley

Organization: La Trobe University

Title-Subject: [Filtered] Post Doc Position Open - ultrastructural biology

Question: A postdoctoral position (Level A/B) is available to work on ultrastructural biology in a new ARC Centre of Excellence for Coherent X-ray Science. ( {http://www.coecxs.org/http://www.coecxs.org/). This position is funded by the Australian Research Council as part of the Centre of Excellence for Coherent X-ray Science for research into the use of novel X-ray diffraction techniques to study the cellular architecture of malaria parasite-infected erythrocytes and other biological samples. The project involves adapting sample preparation methods developed for electron microscopy to generate samples suitable for X-ray microscopy and other X-ray imaging techniques. The successful applicant will work with colleagues from the Departments of Biochemistry and Physics, at La Trobe University, and other Centre members to image the ultrastructure of malaria parasites and other samples. The applicant will also prepare samples for cyro-electron microscopy and single particle image reconstruction techniques. Molecular biological manipulation of cells and proteins will also be used in enhancing sample preparation.

Further information, contact:
Prof. Leann Tilley. Department of Biochemistry, La Trobe University.
Tel: 9479 1375.
Email: {mailto:L.Tilley-at-latrobe.edu.auL.Tilley-at-latrobe.edu.au


Prof Leann Tilley
Phone: 61-3-94791375 Department of Biochemistry
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/labs/tilley/research.html

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From: eschumacher-at-mccrone.com
Date: Thu, 17 Nov 2005 08:40:07 -0600
Subject: [Microscopy] Second Announcement: Student Poster Competition - MMMS Meeting March 24, 2006

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SECOND ANNOUNCEMENT

STUDENT POSTER COMPETITION ANNOUNCEMENT

To be held in conjunction with the March 24, 2006 meeting of the Midwest

Microscopy and Microanalysis Society

Affiliate of the Microscopy Society of America and the Microbeam
Analysis
Society of America

ABSTRACT DEADLINE: Abstracts must be received in electronic format by
5PM, Friday, December 16, 2005.

The first quarter meeting of the Midwest Microscopy and Microanalysis
Society will be held on Friday, March 24, 2006, in the Pancoe Life
Sciences Building, Northwestern University, Evanston, Illinois. A
student poster competition open to undergraduate and graduate students
will be held in
conjunction with the meeting. Posters should illustrate utilization of
microscopy for either biological or materials science study. Prizes
will be awarded as follows:

$300 for 1st place
$200 for 2nd place
$100 for 3rd place

Abstracts must be received in electronic format by 5PM on Friday,
December 16, 2005. To be eligible for a prize, you must be first author
on the poster, and you must be present at the meeting. You are
encouraged to submit your entry as early as possible, as space may be
limited. Abstracts from last year's competition, and an example of the
judging worksheet can be found on the MMMS website:


http://northwestern.edu/bioimaging/MMMS%20Website/index.htm


Further details will be provided upon acceptance of your submission.

Please send the following information to the email address below, and
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Name Phone number
Affiliation Email address
Mailing address

Send to:

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850 Pasquinelli Drive
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Tel: 630-887-7100 Fax: 630-887-7417
Email: eschumacher-at-mccrone.com


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From: Jane.LaGoy-at-bodycote.com
Date: Thu, 17 Nov 2005 09:25:35 -0600
Subject: [Microscopy] viaWWW: question about BSE detectors

Contents Retrieved from Microscopy Listserver Archives
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Matthew - I would contact the SEM manufacturer, JEOL (978-535-5900), and the
EDS manufacturers EDAX (201-529-4880) and PGT (609-924-7310), or check their
websites; I have seen technical articles regarding this authored by their
employees, and there are probably other SEM/EDS companies with helpful
literature as well.

Jane L. LaGoy
Lab Manager/R&D Engineer
978-470-1620 x450
jane.lagoy-at-bodycote.com

BODYCOTE NORTH AMERICA · 155 RIVER STREET · ANDOVER · MASSACHUSETTS ·
01810
TEL: (978) 470-1620 · FAX: (978) 475-2951 · ONLINE: www.bodycote.com




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Email: justgetmeonhere-at-netscape.net
Name: Matthew McCaskill

Organization: none

Title-Subject: [Filtered] question about BSE detectors

Question: Hello all,

I am enrolled in an SEM class, and for this class I have been asked to do a
paper on backscattered electron (BSE) detectors. Specifically, I have to
briefly state what is state-of-the-art now, and then discuss research or
improvements that are occurring right now. Then I am to predict and forecast
the future of the technology; will there be more developments, or have BSE
detectors fully matured technologically?

The point of all this post, however, is that I'm having trouble finding
literature on SEM in general, let alone BSE detectors. Utilizing my school's
library and the internet, I have found at least four articles already, but I
need at least four more. Can anybody recommend me some journal articles or
papers that deal with my topic? Preferably, I'd like for them to be
available as FREE electronic copies (I found a few promising articles
online, but unfortunately I have to pay to view, which is disadvantageous to
poor college students such as myself), but if I can get ahold of the actual
hard copies at my school's library then that should also be enough.

Thanks to all who can help me in this regard, and thanks to all of you for
your time.

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

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From: apnewell-at-ncsu.edu
Date: Fri, 18 Nov 2005 07:23:37 -0600
Subject: [Microscopy] viaWWW: low voltage STEM

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Email: apnewell-at-ncsu.edu
Name: AndyNewell

Organization: AREMC / NC State

Title-Subject: [Filtered] low voltage STEM

Question: I am looking for literature on current developments of SEM-based STEM (low kV) and/or names of predominant researchers that have benefitted recently by using STEM in an SEM for thin sample characterization and microanalysis. Information on other improvements such as aberration corrections that have helped low-voltage STEM or speculation on the next step in low energy STEM development is also welcome.

Many of us graduate students here at NCSU are having difficulty finding publications on development of the technology due to the proprietary nature of such developments, but I am sure there is some fine work going on in the scientififc community. Can anyone help me locate the works of some SEM-based STEM heavy-hitters?

Any assistance available is greatly appreciated.

Many thanks,
Andy Newell
Materials Science & Engineering
NC State University


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From: rbeavers-at-mail.smu.edu
Date: Fri, 18 Nov 2005 11:51:30 -0600
Subject: [Microscopy] Imaging bug fossils

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

Group

l have been trying to look at some bug fossils in rock samples with SEM. In hand specimen these bugs stand out as dark areas against light tan rock material. In the SEM using backscatter they tend to fade into the background which seems to say there is not much organic material left. SEM secondary electron images do not yeild much information either.

Would like to hear any ideas on other techniques people have use to image such samples.

Thanks

Roy Beavers

Southern Methodist University
Department of Geological 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: avklaus-at-amnh.org
Date: Fri, 18 Nov 2005 12:42:24 -0600
Subject: [Microscopy] Imaging bug fossils

Contents Retrieved from Microscopy Listserver Archives
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Hi Roy,

Depending on the composition of the bug fossils, you might have some luck
trying high-resolution CT scanning (microCT). I know of some microCT
scanning successes of mantis fossils with high iron content compared to the
surrounding rock matrix. Although if you are not seeing much average atomic
number difference by BSE imaging, maybe CT would also not work well.

If you need more info on microCT of bugs, please don't hesitate to contact
me offline.

Best regards,

Angela


Angela V. Klaus, Ph.D.
Director, Microscopy and Imaging Facility
American Museum of Natural History
Central Park West and 79th Street
New York, NY 10024 USA

Tel: 212-769-5977
Email: avklaus-at-amnh.org

-----Original Message-----
X-from: rbeavers-at-mail.smu.edu [mailto:rbeavers-at-mail.smu.edu]
Sent: Friday, November 18, 2005 12:52 PM
To: avklaus-at-amnh.org

Group

l have been trying to look at some bug fossils in rock samples with SEM. In
hand specimen these bugs stand out as dark areas against light tan rock
material. In the SEM using backscatter they tend to fade into the background
which seems to say there is not much organic material left. SEM secondary
electron images do not yeild much information either.

Would like to hear any ideas on other techniques people have use to image
such samples.

Thanks

Roy Beavers

Southern Methodist University
Department of Geological 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: oshel1pe-at-cmich.edu
Date: Fri, 18 Nov 2005 14:15:01 -0600
Subject: [Microscopy] RE: Imaging bug fossils

Contents Retrieved from Microscopy Listserver Archives
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Depending on the BSE detector you are using, you might not see much
contrast. We had a 4-quadrant solid-state detector mounted coaxially
with the beam. It does not show much topography when all four quadrants
are summed together. That mode does show the best composition contrast,
but I don't know how much compositional contrast you should expect for
fossils in rock.

Are the fossilizing minerals the same as the host rock? If not, you
should be able to get elemental contrast via an x-ray map image. BTW,
you may need to experiment with the beam voltage depending on the
thickness of your "bugs". A 20kV beam could be blasting all the way
through them.

Still, I would remain hopeful. I know that optical contrast does not
always translate into contrast in the EM, but often there is some way to
tease out contrast in some imaging mode or another.

Warren Straszheim

-----Original Message-----
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Sent: Friday, November 18, 2005 11:53 AM
To: wesaia-at-iastate.edu

Roy,

Two thoughts come to mind. First, if you have access to a low-kV SEM, I'd try imaging at 1 or 1.5 kV with secondaries. You'd more likely differentiate the fossil from the matrix this way. The other thought is: are this organic films? phosphatized? ... ? Meaning, have you tried etching the rocks with weak acetic acid? This often works to bring out fossils (or remove them from the matrix entirely).
It might also be the worst possible thing to do, and should be tried first on a fossil that doesn't matter.

Phil

Philip Oshel
Microscopy Facility Supervisor
Department of Biology
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

Group

l have been trying to look at some bug fossils in rock samples with SEM. In hand specimen these bugs stand out as dark areas against light tan rock material. In the SEM using backscatter they tend to fade into the background which seems to say there is not much organic material left. SEM secondary electron images do not yeild much information either.

Would like to hear any ideas on other techniques people have use to image such samples.

Thanks

Roy Beavers

Southern Methodist University
Department of Geological 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, 18 Nov 2005 14:36:02 -0600
Subject: [Microscopy] Re: Imaging bug fossils

Contents Retrieved from Microscopy Listserver Archives
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What exactly are you trying to accomplish? If you want
morphology, low KV SE should do it. BSE is just going
to show difference in Z. Since the bug is or was organic,
it ought not show much with BSE.

If you want to do morphology and save the specimen, perhaps
doing a replica would work. Use replica resin and then sputter
coat it and then SEM it. Invert the image and you should
have excellent morphology results.

Another idea is CL.

gary g.


At 09:53 AM 11/18/2005, you wrote:



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From: DRK-at-SHCC.org
Date: Fri, 18 Nov 2005 19:15:55 -0600
Subject: [Microscopy] TEM negative scanner

Contents Retrieved from Microscopy Listserver Archives
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Hi,

We've struggled all we can to to keep our Ilford photo processor
working. It's time we entered the digital age. Can anyone suggest a
TEM negative scanner? Ideally it would take a stack of negatives and
feed them into the scanner and produce individual files, so if anyone
can suggest a model that does this I'd be ecstatic. Otherwise, if you
have a scanner that will automatically scan 12 or more negatives,
producing individual files of each, please share your experience.

Many thanks,

doug

Douglas R. Keene
Assistant Investigator
Micro-Imaging Center
Research Department
Shriners Hospital for Children
3101 S.W. Sam Jackson Park Road
Portland, Oregon 97239-3009
voice: 503-221-3434


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From: jmcss-at-aol.com
Date: Sat, 19 Nov 2005 07:57:49 -0600
Subject: [Microscopy] AskAMicroscopist: Middle School - Leaf Stomata Question

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This Question was submitted to Ask-A-Microscopist by (jmcss-at-aol.com)
from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Saturday, November 19, 2005 at 06:46:02
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Email: jmcss-at-aol.com
Name: Mary Hamelin

Organization: Country Day School

Education: 6-8th Grade Middle School

Location: Groton, MA

Title: plants

Question: What is the best way to help the students visualize stomata using a microscope? I have tried clear nail polish to make a relief of the leaf's exterior but didn't have much luck. Is there a way to make good quality, thin sections of plant material with little equipment? I'd like to help the students discover different types of plant cells. Also, what staining, if any would you recommend?

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From: sarj0007-at-unf.edu
Date: Sat, 19 Nov 2005 08:02:13 -0600
Subject: [Microscopy] viaWWW: Joel 35-CF "Smoking"

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Email: sarj0007-at-unf.edu
Name: Jason Saredy

Organization: University of North Florida

Title-Subject: [Filtered] Joel 35-CF

Question: Hi. We are trying to get an old Joel-35CF SEM up and running from a long period of not being used. When turned on there is smoke coming out of the inside of the voltage splitter where, according to the schematics, the transformers are. The smoke doesnít start to come out until about 30-40 seconds after being turned on and its pulling at least 5 amps from a circuit that should only be pulling 3 amps. Anyone knows whether this is an easy to replace part or should we replace the whole power supply. We are doing this out of pocket so prefer to not purchase a new one directly from Joel. If so does anyone have some parts lying around? Or a good website for finding this sort of information? Thanks

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From: jbradley-at-igpp.ucllnl.org
Date: Sat, 19 Nov 2005 08:03:32 -0600
Subject: [Microscopy] viaWWW: A Noran TN5500 program

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Email: jbradley-at-igpp.ucllnl.org
Name: John Bradley

Organization: Lawrence Livermore Laboratory

Title-Subject: [Filtered] SMTF - A Noran TN5500 program

Question: I am looking for a copy of the Noran program "SMTF" (Standardless Metallurgical Thin Film).

I have several hundred TEM/EDX spectra archived on 5.25 inch floppy discs. I also have access to a TN5500 with a 5.25 inch floppy disc drive. I need is a copy of SMTF on a 5.25 inch floppy disc.

I would also very much like to know how these files can be exported from the TN5500.

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From: watson-at-wi.mit.edu
Date: Sat, 19 Nov 2005 08:04:23 -0600
Subject: [Microscopy] viaWWW: Iodine 125/silver enhancement

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Email: watson-at-wi.mit.edu
Name: Nicki Watson

Organization: whitehead Institute

Title-Subject: [Filtered] Iodine 125/silver enhancement

Question: Hi
I am looking for protocols for immunolabeling cultured cells using Iodine 125/silver enhancement techniques.
I think this is a rather old fashion method, but I have a professor interested in it for a specific project. We would also be interested in
the opinions of you experts as to the functionality of this technique.Do you prefer gold, or peroxidase, quantum dots ? Does the iodine 125 have any special features that make it a great choice? Any input would be great!
thanks for your input
Nicki Watson

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From: baskin-at-bio.umass.edu
Date: Sat, 19 Nov 2005 10:41:33 -0600
Subject: [Microscopy] Re: AskAMicroscopist: Middle School - Leaf Stomata

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

Hi Mary,
I am not an expert in this area but I can suggest a couple of
things to try.

First, check out Project Micro, this has a ton of useful information
about microscopes in education, for all levels.
http://microscopy.org/ProjectMICRO/

Second, don't forget to see what you can see with a good hand lens,
for example a jewler's loupe. This shows a great deal (though not
usually cells) and is a great intermediate between the eye and the
microscope. Lots of leaves (flowers, etc) make elaborate hairs and
things, which look great even with modest magnification.

Third, remember that in many (most?) plants, stomata are on the
abaxial side of the leaf (ie, towards the ground).

Fouth. A trick popular with stomatal researchers is to make an
epidermal peel. You take a tweezers and grab near the midvein and
carefully pull toward the leaf margin. You can often remove a flap of
epidermis. Put that in some water on a slide under a coverslip and
you should be able to see stomata even in brighfield. Supposedly
Vicia faba (bean) is easy to peel. But note that for your purpose,
even a small (few mm square) area should suffice. It might be fun for
your class to try some different plants and see which peel better?

Fifth. If you do get good at peeling, you can even try some things to
open or close the stomata. Add some CaCl2 to the water where the
peels are, or sodium bicarbonate. See what happens. Manipulate the
light environment of the plants before you make peels (keep some
plants in the dark, others in bright light). This could be beyond
what you have in mind, but I thought I'd mention it just in case.

Have fun

Hope this helps,
Tobias Baskin

}
}
} Email: jmcss-at-aol.com
} Name: Mary Hamelin
}
} Organization: Country Day School
}
} Education: 6-8th Grade Middle School
}
} Location: Groton, MA
}
} Title: plants
}
} Question: What is the best way to help the students visualize
} stomata using a microscope? I have tried clear nail polish to make a
} relief of the leaf's exterior but didn't have much luck. Is there a
} way to make good quality, thin sections of plant material with
} little equipment? I'd like to help the students discover different
} types of plant cells. Also, what staining, if any would you
} recommend?
}

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

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From: cbalane-at-wesleyan.edu
Date: Sat, 19 Nov 2005 13:35:26 -0600
Subject: [Microscopy] gold conjugation with fluorescent BSA

Contents Retrieved from Microscopy Listserver Archives
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Hi everyone,
I am trying to conjugate my colloidal gold particles (0.8 nm and 6 nm
particle sizes) with fluoresceinated BSA. I have tried to perform
extensive literature search for detailed protocols but most of what I
found were either for specific proteins (like IgG or Protein A) or general
recipes that don't really say much about what I should do.

I know that we have experts out there who have done such procedures (or
others that may be very similar) in the past; any advice/information would
be greatly appreciated.

Have a happy thanksgiving!

Sincerely,
Carlo

---

Carlo Franco Bolivar Balane

Box 4058, 222 Church Street, or Wolfe Laboratory
Wesleyan University Station Rm. 157, HA Laboratories
Middletown, CT, 06459-4058 Wesleyan University
phone: 1.860.759.2830 phone: 1.860.685.3275


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From: mike.reedy-at-cellbio.duke.edu
Date: Sat, 19 Nov 2005 17:15:13 -0600
Subject: [Microscopy] Re: TEM negative scanner

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

Hi Doug,
The $350-500 Epson and Canon flatbeds seem hard to beat for price and
resolution, but don't have film autofeeders or multi-holders for 3.25
x 4" negatives; you must fit one negative at a time into the 4"x5"
holder. I'm not sure jamproof autofeeders exist.

600 dpi scans will serve most workprint needs (600 scans 29 sec or
less with new EPSON 4990 PRO, says Amazon). Keep the film! for
selected re-scan when needing greater enlargement. My betters advise
that it is better (more dynamic range/) to scan as a transparency and
invert rather than as a negative. Make excellent workprints fast on
standard paper on an HP 4100 LaserJet set for 150 lpi. (exceeds
printer spec of 1200 dpi, I know, but it works).

For general info look at http://scantips.com/ .

For additional general info look over many comments and viewpoints at
http://www.macintouch.com/scanners05.html (also parts 4 amd 6)
Note Steve Pucci's comments on batch-feed film scanners under Aug
25, 2005 entries.
Check out his preferred Canon 9950 scanner for its Customer Reviews
comments at Amazon.com; then see also the Epson Perfection 4870 PRO
Customer Reviews. The newest Epson, 4990 PRO, has no Amazon reviews
yet, but see this and other scanners, including Microtek, reviewed at
CNET:
http://reviews.cnet.com/Epson_Perfection_4990_Pro/4514-3136_7-31320383.html
(latter site also has reviews and customer comments for the other top
scanners).

-mike reedy


At 7:20 PM -0600 11/18/05, DRK-at-SHCC.org wrote:
} ----------------------------------------------------------------------------
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--
-mike reedy-

************************
Michael K. Reedy, M.D.
Duke Univ. Med. Center
Dept. Cell Biology, Box 3011 (for U.S. Mail)
458 Alex Sands Bldg, Research Dr (courier)
Durham, NC 27710

Office 919-668-2534
Lab 919-684-5674
Fax 919-681-9929
mike.reedy-at-cellbio.duke.edu
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From: rpowell-at-nanoprobes.com
Date: Sun, 20 Nov 2005 09:10:32 -0600
Subject: [Microscopy] viaWWW: gold conjugation with fluorescent BSA

Contents Retrieved from Microscopy Listserver Archives
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This Question/Comment was submitted to the Microscopy Listserver
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Email: rpowell-at-nanoprobes.com
Name: Richard D. Powell

Organization: Nanoprobes, Incorporated

Title-Subject: [Filtered] Re: [Microscopy] gold conjugation with fluorescent BSA

Question: Hello Carlo:

Most colloidal gold conjugation protocols are for antibodies or other targeting agents. BSA is used as a stabilizer in these reactions, but because conjugation of the targeting agent takes priority, the reaction conditions are usually optimized for conjugating to this rather than the BSA. However, you don't need to make many changes - the procedure is similar for most colloidal gold conjugations.

Optimum colloidal gold conjugation is usually done at a pH at or just above the pI of the protein you are conjugating. From a quick search, the pI of BSA is about 4.7, so a pH of about 5 to 5.2 would be good for conjugation. Usually you would dialyze the protein into a dilute buffer at this pH or deionized water; use a buffer that does not flocculate colloidal gold (for example 0.002 M citrate, pH 5). Adjust the pH of the colloidal gold sol to the same pH using dilute H3PO4; then add the minimum stabilizing amount of fluorescein-BSA to the gold (you can determine this from titration; a procedure is given in the link below), stir for 2 minutes, then add a further 10% of BSA and 0.1% carbowax (a special form of 20,000 MW polyethylene glycol), stir 10 - 15 minutes more.

A good procedure for antibodies is given in this link. To adapt for albumin, all you need to do is to change the pH for the titration and conjugation to one more suited to BSA (adjust pH with H3PO4 if you need to lower rather than raise it):

http://www.researchd.com/gold/gold8.htm

All glassware must be scrupulously clean. Glass and plastic containers and stirrers should be cleaned in aqua regia, thoroughly washed in deionized water, and siliconized.

Once you have prepared the conjugate, you should store at a pH several units away from the PI, as this helps stabilize the conjugate. Spin down and resuspend in 0.02 M Tris-HCl, pH 8.2 (the usual buffer for storing colloidal gold conjugates).

Before doing this, though, you should be aware that fluorescence will likely be completely quenched if you conjugate fluorescein-streptavidin to 6 nm gold - gold is a very efficient absorber for resonance energy transfer, and other mechanimsms may also be present that will quench the fluorescence even more than energy transfer alone. We have explained this, and Albrecht and co-workers have observed it in practice when making combined fluorescent and 6 nm gold-labeled antibodies:

(1) Powell, R. D.; Halsey, C. M. R., and Hainfeld, J. F.: Combined fluorescent and gold immunoprobes: Reagents and methods for correlative light and electron microscopy. Microsc. Res. Tech., 1998, 42, 2.

(2) Kandela, I. K.; Meyer, D. A.; Oshel, P. E.; Rosa-Molinar, E., and Albrecht, R. M.: Fluorescence Quenching by Colloidal Heavy Metals: Implications for Correlative Fluorescence and Electron Microscopy Studies. Microsc. Microanal., 9, (Suppl. 2: Proceedings); Piston, D.; Bruley, J.; Anderson, I. M.; Kotula, P.; Solorzano, G.; Lockley, A., and McKernan, S. (Eds.); Cambridge University Press, New York, NY, 2003, 1194CD.

Hope this is helpful,

Rick Powell

*****************************************************************************************
Richard D. Powell
www.nanoprobes.com
NANOPROBES, Incorporated
*****************************************************************************************

Hi everyone,
I am trying to conjugate my colloidal gold particles (0.8 nm and 6 nm
particle sizes) with fluoresceinated BSA. I have tried to perform
extensive literature search for detailed protocols but most of what I
found were either for specific proteins (like IgG or Protein A) or general
recipes that don't really say much about what I should do.

I know that we have experts out there who have done such procedures (or
others that may be very similar) in the past; any advice/information would
be greatly appreciated.

Have a happy thanksgiving!

Sincerely,
Carlo

---

Carlo Franco Bolivar Balane

Box 4058, 222 Church Street, or Wolfe Laboratory
Wesleyan University Station Rm. 157, HA Laboratories
Middletown, CT, 06459-4058 Wesleyan University
phone: 1.860.759.2830 phone: 1.860.685.3275


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

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From: rpowell-at-nanoprobes.com
Date: Sun, 20 Nov 2005 09:28:51 -0600
Subject: [Microscopy] viaWWW: gold conjugation with fluorescent BSA

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.org/MicroscopyListserver/MLFormMail.html
---------------------------------------------------------------------------
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Email: rpowell-at-nanoprobes.com
Name: Richard D. Powell

Organization: Nanoprobes, Incorporated

Title-Subject: [Filtered] Re: [Microscopy] gold conjugation with fluorescent BSA

Question: Hello Carlo:

Most colloidal gold conjugation protocols are for antibodies or other targeting agents. BSA is used as a stabilizer in these reactions, but because conjugation of the targeting agent takes priority, the reaction conditions are usually optimized for conjugating to this rather than the BSA. However, you don't need to make many changes - the procedure is similar for most colloidal gold conjugations.

Optimum colloidal gold conjugation is usually done at a pH at or just above the pI of the protein you are conjugating. From a quick search, the pI of BSA is about 4.7, so a pH of about 5 to 5.2 would be good for conjugation. Usually you would dialyze the protein into a dilute buffer at this pH or deionized water; use a buffer that does not flocculate colloidal gold (for example 0.002 M citrate, pH 5). Adjust the pH of the colloidal gold sol to the same pH using dilute H3PO4; then add the minimum stabilizing amount of fluorescein-BSA to the gold (you can determine this from titration; a procedure is given in the link below), stir for 2 minutes, then add a further 10% of BSA and 0.1% carbowax (a special form of 20,000 MW polyethylene glycol), stir 10 - 15 minutes more.

A good procedure for antibodies is given in this link. To adapt for albumin, all you need to do is to change the pH for the titration and conjugation to one more suited to BSA (adjust pH with H3PO4 if you need to lower rather than raise it):

http://www.researchd.com/gold/gold8.htm

All glassware must be scrupulously clean. Glass and plastic containers and stirrers should be cleaned in aqua regia, thoroughly washed in deionized water, and siliconized.

Once you have prepared the conjugate, you should store at a pH several units away from the PI, as this helps stabilize the conjugate. Spin down and resuspend in 0.02 M Tris-HCl, pH 8.2 (the usual buffer for storing colloidal gold conjugates).

Before doing this, though, you should be aware that fluorescence will likely be completely quenched if you conjugate fluorescein-streptavidin to 6 nm gold - gold is a very efficient absorber for resonance energy transfer, and other mechanimsms may also be present that will quench the fluorescence even more than energy transfer alone. We have explained this, and Albrecht and co-workers have observed it in practice when making combined fluorescent and 6 nm gold-labeled antibodies:

(1) Powell, R. D.; Halsey, C. M. R., and Hainfeld, J. F.: Combined fluorescent and gold immunoprobes: Reagents and methods for correlative light and electron microscopy. Microsc. Res. Tech., 1998, 42, 2.

(2) Kandela, I. K.; Meyer, D. A.; Oshel, P. E.; Rosa-Molinar, E., and Albrecht, R. M.: Fluorescence Quenching by Colloidal Heavy Metals: Implications for Correlative Fluorescence and Electron Microscopy Studies. Microsc. Microanal., 9, (Suppl. 2: Proceedings); Piston, D.; Bruley, J.; Anderson, I. M.; Kotula, P.; Solorzano, G.; Lockley, A., and McKernan, S. (Eds.); Cambridge University Press, New York, NY, 2003, 1194CD.

Hope this is helpful,

Rick Powell

*****************************************************************************************
Richard D. Powell
www.nanoprobes.com
NANOPROBES, Incorporated
*****************************************************************************************

Hi everyone,
I am trying to conjugate my colloidal gold particles (0.8 nm and 6 nm
particle sizes) with fluoresceinated BSA. I have tried to perform
extensive literature search for detailed protocols but most of what I
found were either for specific proteins (like IgG or Protein A) or general
recipes that don't really say much about what I should do.

I know that we have experts out there who have done such procedures (or
others that may be very similar) in the past; any advice/information would
be greatly appreciated.

Have a happy thanksgiving!

Sincerely,
Carlo

---

Carlo Franco Bolivar Balane

Box 4058, 222 Church Street, or Wolfe Laboratory
Wesleyan University Station Rm. 157, HA Laboratories
Middletown, CT, 06459-4058 Wesleyan University
phone: 1.860.759.2830 phone: 1.860.685.3275


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

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From: heckman-at-bgnet.bgsu.edu
Date: Sun, 20 Nov 2005 14:12:38 -0600
Subject: [Microscopy] Re: gold conjugation with fluorescent BSA

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

Dear Carlo-

Gold particles (and in fact, most clean surfaces of a noble metal,
will bind to any type of protein. The binding is apparently not
covalent but it is very tight. If you follow a procedure for binding
IgG or Protein A, but what you supply is only BSA, the BSA will bind.
BSA does not adhere to surfaces as tightly as many other proteins.
There is a lot of literature on the adhesion, look particularly for
articles by Andrade who was at University of Utah. If you
cross-search adhesion and the professor's name, you should be able to
find them.
Carol


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Professor of Biological Sciences
Director, Center for Microscopy & Microanalysis
Bowling Green State University, Bowling Green, OH 43403
fax: (419) 372-2024 email: heckman-at-bgnet.bgsu.edu
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___________________________________________________________________________

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From: Rosey.VanDriel-at-csiro.au
Date: Sun, 20 Nov 2005 16:22:48 -0600
Subject: [Microscopy] TEM: HM20 problems

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

Dear Listers,
I have been using HM20 Monostep (premixed) resin for freeze substitution
for a couple of years, and have recently encountered problems with batch
variability. Sarah Ellis of the Peter MacCallum Cancer Clinic in
Melbourne has recently experienced similar problems.
The main problem is incomplete or variable infiltration, resulting in
polymerised resin pulling away from the edge of tissue blocks or cells,
leaving gaps and making sections hard to cut and to view. In some cases
there are gaps around larger organelles within the cells. Another batch
frosted badly at -45 degrees C.

My question to listserver members is whether anyone else is having these
variability problems. More importantly, does anyone have a solution?

Thanks,
Rosey van Driel


Rosemary van Driel
Electron Microscopy
New and Emerging Zoonotic Diseases
Australian Animal Health Laboratory
CSIRO Livestock Industries
Private Bag 24
Geelong Vic 3220
Australia

International Phone: +61 3 5227 5209
International Fax: +61 3 5227 5555
email: Rosey.VanDriel-at-csiro.au


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From: mcauliff-at-umdnj.edu
Date: Mon, 21 Nov 2005 10:32:48 -0600
Subject: [Microscopy] Re: TEM negative scanner

Contents Retrieved from Microscopy Listserver Archives
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I'm afraid that such a scanner would be very expensive. To scan the
full frame of a 3.25x4 inch EM negative you will need a 4x5 film
scanner, not an inexpensive item.
I suggest two alternatives. 1. A good flatbed scanner will be 1/10
the cost of a 4x5 film scanner. 2. Put your negatives on a light box
and photograph them with a digital camera. Most will focus very close,
then invert the image (negative to postitive) in PhotoShop.

Geoff

DRK-at-SHCC.org wrote:

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From: Rosey.VanDriel-at-csiro.au
Date: Mon, 21 Nov 2005 22:34:48 -0600
Subject: [Microscopy] TEM: HM20 problems: summary

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

Dear listers,
It would seem that our problems with Monostep HM20 may be due to aged or
contaminated resin.

The Monostep was supplied by a company which had its own labels on
otherwise unlabelled bottles. I have since learnt that the product is
shipped from the manufacturer with a lot number and expiry date. The
expiry dates are not on the product I received, which suggests there may
have been some rebottling or aging of the product. Interestingly, the
manufacturer was unable to trace the lot numbers printed on the
supplier's labels. The supplier refuses to entertain the thought that a
problem exists with the resin we recieved.

I have also discovered that resins ( and I don't know what else) can go
through 3 resellers before they reach us in our labs. So buyer beware!

I have traced a more direct source of supply, so I hope that the problem
is solved.

Thanks especially to the US commercial list subsciber who spared no
effort in helping track down the likely sources of the problem, I'll get
Alex to buy you a beer in Sydney in February. Thanks also to the other
supply companies in Australia who helped. I'd like to name the helpful
ones, but I don't think it's allowed.

Rosey



Rosemary van Driel
Electron Microscopy
New and Emerging Zoonotic Diseases
Australian Animal Health Laboratory
CSIRO Livestock Industries
Private Bag 24
Geelong Vic 3220
Australia

International Phone: +61 3 5227 5209
International Fax: +61 3 5227 5555
email: Rosey.VanDriel-at-csiro.au


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From: brakenho-at-science.uva.nl
Date: Tue, 22 Nov 2005 09:22:03 -0600
Subject: [Microscopy] FOM2006, Abstracts, Registration, Perth, Australia, April 9-12,

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

FOCUS ON MICROSCOPY 2006, Perth, Australia, April 9-12, 2006
19th International Conference on 3D Image Processing in Microscopy
18th International Conference on Confocal Microscopy

2nd Announcement:

Dear Colleagues,

Abstracts for oral and poster presentations can now be submitted
preferably through the conference website:
http://www.focusonmicroscopy.org
where also the conference registration has opened and hotel
information is available.

Deadline for abstract submission: Jan. 9, 2006.
Note: this is earlier than in previous conferences!

The program will start on Sunday April 9, around 18 hours with an
opening symposium followed by a welcome reception.

After the successful FOM2005 conference in Jena, the next conference Focus
on Microscopy 2006 will take place in Perth, Western Australia, April
9-12, 2006. FOM2006 is the next in a series of unique interdisciplinary
meetings on advanced multidimensional light microscopy and image
processing. The conference will be hosted by the University of Western
Australia in Perth and held in the beautiful Esplanade Hotel in the
historic waterfront suburb of Perth, Fremantle.

Focus on Microscopy 2006 is the continuation of a conference series
presenting the latest innovations in optical microscopy and its
applications in biology, medicine, material science, and information
storage. 3D optical imaging and related theory are traditional subjects
for the conference. The conference series is also known for covering the
rapid development of advanced fluorescence labeling techniques for the
confocal andmulti-photon 3D imaging of -live- biological specimens.
This year, in addition, special attention will be given to imaging in
thick tissues.

Abstracts for contributions are invited and can now be submitted through
the website:

www.FocusOnMicroscopy.org

where further information on the present and previous FOM conferences can
be found.

Important dates:

Deadline for the submission of abstracts: January 9, 2006
Draft program available on the web: January 23, 2006
at website www.FocusOnMicroscopy.org
Deadline for early registration: February 20, 2006

Welcoming you to beautiful Perth for the FOM2006
conference and exhibition.

On behalf of the organizing committee,

David Sampson,
University of Western Australia, Perth, Australia

Fred Brakenhoff
Swammerdam Institute for Life Sciences, University of Amsterdam, The
Netherlands

E-mail: info2006-at-FocusOnMicroscopy.org

Web: www.FocusOnMicroscopy.org




==============================Original Headers==============================
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From: Kerstin.Brismar-at-vv.slu.se
Date: Tue, 22 Nov 2005 10:09:05 -0600
Subject: [Microscopy] SEM - Cryo system for sale

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

Cryo system for SEM, Oxford CT 1500B, bought in
1996, in excellent condition, used only three
times, will be sold for a very reasonable price.
A good opportunity to try the cryo technique, or
to get spare parts for an existing system. Don't
hesitate to ask for more information!


******************************
Kerstin Brismar
B.Sc., Research engineer, Photographer
Dept. of Crop Science
SLU (Swedish University of Agricultural Sciences)
P.O. Box 44 (Delivery: Växtskyddsvägen 1)
SE-230 53 Alnarp, Sweden
Phone: +46 40 41 55 05
Fax: +46 40 41 55 19
E-mail: Kerstin.Brismar-at-vv.slu.se
******************************



==============================Original Headers==============================
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From: mike.reedy-at-cellbio.duke.edu
Date: Tue, 22 Nov 2005 15:49:33 -0600
Subject: [Microscopy] TEM negative scanner

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

Michael-
I checked your question with my Source, because I remembered only
that I had long shared your viewpoint, and that he had changed my
mind, but how....? So he reminded me . (The "they" he says set a
smaller DR aperture for negative scans than for positive transparency
scans are not Kodak, but scanner-makers in general, who rely on
Kodak's loooong experience):

"I know it is counterintuitive but it is true and can be easily
tested. In their haste to make scanning "better" they make and
assumption based on loooong experience. Kodak discovered that most
negatives have a transmission of 54% +- 5 %. They therefore set a
smaller aperture in the dynamic range to get a better scan. The
problem is that scientific imaging has a much wider need. Think
brightfield microscopy with 98% transmission or darkfield with 2%.
Because positive transparencies can have dark and light background
they open up the dynamic range to get better scans. In most scanners
scanning as a positive transparency and inverting will give you a
much better histogram than scanning as a negative.

Epson is ONLY scanner that I recommend; this is because the actual
calibrated resolution (I calibrated it myself) is 2000 dpi (4800dpi
claimed). This is half the possible resolution of film, that can
only be beaten with a 15,000 dpi drum scanner.

Our solution to a lack of film feeding is to put a scanner on every
desk. Scanning can be done while doing email/reading/ etc"

My own thought is-- if there were no such difference, why would
scanner controls continue to offer both options? ---- given that
"invert" has been available with a keystroke for some time, at least
since PhotoShop 2.0. I can think of possible simple tests, but I've
yet to perform any.

-mike reedy-


} Mike Reedy writes ...
}
}
} } [...]
} } My betters advise that it is better (more dynamic range/) to
} } scan as a transparency and invert rather than as a negative.
} } [...]
}
} I don't believe this is correct. How is better DR related to inverting the
} grayscale?
}
} genuinely :o)
} michael shaffer
}
} SEM/MLA Laboratory Coordinator
} (709) 737-6799 (ofc)
} (709) 737-6790 (lab)
} (709) 737-6193 (FAX)
} {www.mun.ca/creait/maf/}
}
} Inco Centre
} c/o Memorial University
} 230 Elizabeth Avenue
} P.O. Box 4200
} St. John's, NL A1C 5S7


--
-mike reedy-

************************
Michael K. Reedy, M.D.
Duke Univ. Med. Center
Dept. Cell Biology, Box 3011 (for U.S. Mail)
458 Alex Sands Bldg, Research Dr (courier)
Durham, NC 27710

Office 919-668-2534
Lab 919-684-5674
Fax 919-681-9929
mike.reedy-at-cellbio.duke.edu
http://note.cellbio.duke.edu/Faculty/Research/Reedy.html

==============================Original Headers==============================
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12, 20 -- From: Mike Reedy {mike.reedy-at-cellbio.duke.edu}
12, 20 -- Subject: RE: [Microscopy] Re: TEM negative scanner
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From: gcc-at-couger.com
Date: Tue, 22 Nov 2005 19:01:48 -0600
Subject: [Microscopy] Re: TEM negative scanner

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

Mike,

As you approach the resolution of film what problems do you see with
scanning the image from a random pattern of grain to the regular patter of
pixels. It obvious that straight line at angles other than the XY pattern
of the pixels are a problem but there are not that many straight lines in
most images.

With large pixels and small grain there is a loss of contrast does that
hold up as the pixel and grain size converge? As we are one or two
doublings of scanning resolution away from the point that digital is as
good or better than film the differences become more interesting.

With the drum scanners we have today one could scan in Kodak Tri X shot at
ISO 800 developed in HC 110 dilution B and be able to over sample the
grain on the negative for experemanal work. I think there are some photo
journalist who still shoot this combination. It has a unique look on news
print that some like.

I don't think there is anything standing in the way of higher resolution
drum scanners but the size of the image being 4 time larger for every
doubling in resolution and the computer power increasing by the square of
the time it takes to do operations. Combine that with the cost of making a
scanner to those tolerances and there is probably not a market for it.

Gordon
Gordon Couger

I collect links on information related to light microscopes.
www.couger.com/microscope/links/gclinks.html
Please forward anything you think might be useful to others.
Microscope Documentation is at www.science-info.org

mike.reedy-at-cellbio.duke.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
} ----------------------------------------------------------------------------
}
} Michael-
} I checked your question with my Source, because I remembered only
} that I had long shared your viewpoint, and that he had changed my
} mind, but how....? So he reminded me . (The "they" he says set a
} smaller DR aperture for negative scans than for positive transparency
} scans are not Kodak, but scanner-makers in general, who rely on
} Kodak's loooong experience):
}
} "I know it is counterintuitive but it is true and can be easily
} tested. In their haste to make scanning "better" they make and
} assumption based on loooong experience. Kodak discovered that most
} negatives have a transmission of 54% +- 5 %. They therefore set a
} smaller aperture in the dynamic range to get a better scan. The
} problem is that scientific imaging has a much wider need. Think
} brightfield microscopy with 98% transmission or darkfield with 2%.
} Because positive transparencies can have dark and light background
} they open up the dynamic range to get better scans. In most scanners
} scanning as a positive transparency and inverting will give you a
} much better histogram than scanning as a negative.
}
} Epson is ONLY scanner that I recommend; this is because the actual
} calibrated resolution (I calibrated it myself) is 2000 dpi (4800dpi
} claimed). This is half the possible resolution of film, that can
} only be beaten with a 15,000 dpi drum scanner.
}
} Our solution to a lack of film feeding is to put a scanner on every
} desk. Scanning can be done while doing email/reading/ etc"
}
} My own thought is-- if there were no such difference, why would
} scanner controls continue to offer both options? ---- given that
} "invert" has been available with a keystroke for some time, at least
} since PhotoShop 2.0. I can think of possible simple tests, but I've
} yet to perform any.
}
} -mike reedy-
}
}
}
} } Mike Reedy writes ...
} }
} }
} }
} } } [...]
} } } My betters advise that it is better (more dynamic range/) to
} } } scan as a transparency and invert rather than as a negative.
} } } [...]
} }
} } I don't believe this is correct. How is better DR related to inverting the
} } grayscale?
} }
} } genuinely :o)
} } michael shaffer
} }
} } SEM/MLA Laboratory Coordinator
} } (709) 737-6799 (ofc)
} } (709) 737-6790 (lab)
} } (709) 737-6193 (FAX)
} } {www.mun.ca/creait/maf/}
} }
} } Inco Centre
} } c/o Memorial University
} } 230 Elizabeth Avenue
} } P.O. Box 4200
} } St. John's, NL A1C 5S7
}
}
}


==============================Original Headers==============================
9, 21 -- From gcc-at-couger.com Tue Nov 22 19:01:47 2005
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9, 21 -- Subject: Re: [Microscopy] TEM negative scanner
9, 21 -- References: {200511222224.jAMMOoPo007503-at-ns.microscopy.com}
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From: microfrog9-at-hotmail.com
Date: Tue, 22 Nov 2005 20:29:08 -0600
Subject: [Microscopy] [Filtered] MicroscopyListserverviaWWW: Phase Plate

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/MicroscopyListserver/MLFormMail.html
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Email: microfrog9-at-hotmail.com
Name: Lisa Smith

Organization: University of Toronto

Title-Subject: [Filtered] Phase Contrast Microscope - Phase Plate

Question: What is the equation used to calculate the thickness of the etched region of the phase ring?

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

==============================Original Headers==============================
6, 12 -- From zaluzec-at-microscopy.com Tue Nov 22 20:29:08 2005
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6, 12 -- Subject: [Filtered] MicroscopyListserverviaWWW: Phase Plate
6, 12 -- Content-Type: text/plain; charset="us-ascii"
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From: psneeley-at-xmission.com
Date: Tue, 22 Nov 2005 20:29:51 -0600
Subject: [Microscopy] [Filtered] MicroscopyListserverviaWWW: AO Spencer Series 4

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: psneeley-at-xmission.com
Name: Steve Neeley

Title-Subject: [Filtered] Looking for an AO Spencer Series 4 Microscope Reference Manual

Question: I have an AO Spencer Series 4 Microscope and need the Reference Manual. I have the Catalog, the Phase Catalog, and the Phase manual, but I need the actual reference manual that would have been provided for the scope by AO Spencer. These scopes were made in the 1950's and are blue/gray in color (Earlier scopes were the black Spencers and AO Spencers, later scopes were the Gray Series 10, etc.)

These microscope were heavily used at Colleges and Universities (many are still on the shelves, in labs, and in storage at least) and I'm hoping that someone has a copy of the manual.

If you have one, please contact me.

Thank you.

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

==============================Original Headers==============================
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8, 13 -- From: psneeley-at-xmission.com (by way of MicroscopyListserver)
8, 13 -- Subject: [Filtered] MicroscopyListserverviaWWW: AO Spencer Series 4
8, 13 -- Microscope Reference Manual
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From: pekysar-at-ucdavis.edu
Date: Tue, 22 Nov 2005 20:30:19 -0600
Subject: [Microscopy] viaWWW: Identifying motor endplates in thick sections

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.org/MicroscopyListserver/MLFormMail.html
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Email: pekysar-at-ucdavis.edu
Name: Pat Kysar

Organization: University of California, Davis, School of Medicine,Pathology

Title-Subject: [Filtered] Identifying motor endplates in thick sections

Question: Hi all,
We have a pathologist who is conducting a study which necessitates the need to locate motor endplates in resin embedded muscle tissue. He would like to sucessfully locate them in thick sections so we can accurately cut thin sections which includes the area of interest.
Does anyone have a protocol for staining the endplates on the thick sections (epoxy resin embedded) to facilitate a more accurate identification of these structures? The pathologist has tried finding the areas on Methylene blue/Azure B stained sections with a very low rate of success and desires a more accurate method.
Any help would be appreciated.
Thanks,

Pat Kysar
EM Lab
University of California, Davis
School of Medicine, Pathology
1 Shields
Davis, CA 95616
pekysar-at-ucdavis.edu

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

==============================Original Headers==============================
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From: gary-at-gaugler.com
Date: Tue, 22 Nov 2005 23:21:45 -0600
Subject: [Microscopy] TEM negative scanner

Contents Retrieved from Microscopy Listserver Archives
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Diatribe aside....

What is your suggestion for a scanner?

Gary g.




At 05:03 PM 11/22/2005, you wrote:



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From: c.jeffree-at-ed.ac.uk
Date: Wed, 23 Nov 2005 02:47:49 -0600
Subject: [Microscopy] TEM negative scanner

Contents Retrieved from Microscopy Listserver Archives
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Depends on your objectives, I suspect.
If your objective is optimum combination of quality and versatility at the
lowest
cost then you can't go past Epson Photo scanners.
Epson 4870 Photo can generate embarassingly large files from an EM negative
which contain more detail than you can see on an 8"x10" photograph, and the
quality
of scans of 35mm transparencies can be good, more than adequate for 90% of
work in
scientific publishing. Having said that, photo buffs and EM people who are
looking for highest fidelity will
quickly notice limitations. DMax of 3.8 on 4870 (4.0 on 4990) looks good,
but in practice flare from the uncoated glass
platen compromises this performance. Images with extreme contrast (e.g. hard
black areas against pure white)
show visible and objectionable bleeding of light and colour into the dark
areas. I find illumination is not uniform, so that large images with
large areas of uniform density reproduce with density variation that is hard
to correct.
The 4870 is not really capable of resolving the grain of my transparencies,
and introduces a characteristic artefactual speckling or peppering of pixels
correlated with high-frequency information that can be objectionable when
images are enlarged to display sizes.
For this reason, I regard many of my scans of slides as adequate for
printing A4, but below par for archiving. If these things will worry you,
then you should probably
look at a dedicated film scanner such as one of the Nikon Coolscans. They
cost between 2 and 10 times the price,
but you get what you pay for.

Chris

----- Original Message -----
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From: keith.morris-at-ucl.ac.uk
Date: Wed, 23 Nov 2005 03:43:40 -0600
Subject: [Microscopy] TEM negative scanner

Contents Retrieved from Microscopy Listserver Archives
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Hi

As the chat on scanners is continuing I'll add my thoughts.

We use an 'old' semi-pro Agfa flatbed scanner that cost five thousand pounds
in its day - which runs on an Apple, as a Mac person bought it and
most hi-end graphics were then MAC orientated. Unfortunately Agfa,
Heidelberg and Fuji seem to have abandoned the pro scanner market, but the
hi-res scanning of many large negatives manually wasn't a problem for us, as
the scanner captures the images quite quickly and often we didn't require
full resolution for subsequent image analysis - in fact most time is spent
getting the images from Apple to IBM PC's. Apparently the cost of buying a
digital camera add-on for our units TEM would be around £70k with 2k x 2k
pixels and around £20k for 1k x 1k pixels (although our EM dept. has just
decided to go down the £70k route and dispense with film completely).

Creo is one of the few companies that I know that still make this type of
flat bed pro scanner, but I've no experience of their products. A scanner
that can take lots of 4x5" negatives across the whole of its scan area and
scan at 5000 dpi used to cost £10k +, and I have to admit I'm quite sure
what, if anything, has replaced them. As we have the original negative
anyway, 1600 dpi or less is fine for our digitising (mostly for subsequent
image analysis or incorporation into documents). Our scanner is naturally
used for slides, film, photo and paper copying as well.

You can still buy a few cheap sub £200 scanners with a light bulb area
larger than 35mm for transpancies, but all the best cheap ones seem fixed to
35mm slides/negatives. As mentioned by others, probably something like the
£750 Epson Expression 1680 Pro with its A4 transparency adapter would suite
for manual scanning - the A4 Transparency Unit "provides the capability to
scan multiple positive & negative films of up to 5"x4" in size" (the 1680
has 1600 x 3200dpi optical resolution). However we haven't got one to say
whether it is as good as our old Agfa (it's certainly a lot cheaper but
has similar specs). Things like hardware Digital ICE are probably less
useful as our negatives are in good condition (although it works well on my
home Benq Scanwit 2740S slide scanner that's used to scan very old colour
slides of the family). Negative/slide scanners do seem to have real problems
with grain size on my recent colour ASA 400 [and above] negatives generating
optical effects, but fortunately that isn't a problem we see with our TEM
negatives or my old Kodak/Agfa/Perutz 50 to 100 ASA slides from the
50s-70's.

Keith

----------------------------------------------------------
Dr Keith J Morris
Imaging Facilities Manager
Cell Biology Division
Institute of Ophthalmology
University College London
11-43 Bath Street
London EC1V 9EL

Tel: 020 7608 4050
Fax: 020 7608 4034
email: keith.morris-at-ucl.ac.uk
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From: keith.morris-at-ucl.ac.uk
Date: Wed, 23 Nov 2005 04:18:11 -0600
Subject: [Microscopy] TEM negative scanner

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

Hi

By the way, If you are 'archiving' the TEM negatives in maximum resolution
and possibly dumping the negative, I would buy the Nikon LS 900D - it's
£2,500 but I'm sure the smaller Nikon scanners are all 35mm only. Compared
to a digital camera on the TEM I suppose the price isn't bad. It can't scan
photo's or paper though. From my experience of scanning slides it can be a
daunting task working through an old archive at 3 minutes per photo just to
scan the TEM negative, but for new output that may be fine. It would be
great for my home slides though at 40s for 35mm.

Nikon details:

The SUPER COOLSCAN 9000 ED's multi-format capability is specifically
designed for imaging professionals. Scanning is possible for 120/220, 35mm,
6 x 7, 6 x 9 positives, 16mm, electron microscope and other film formats.
The 9000 ED's large-diameter Scanner Nikkor ED lens, 3-line CCD image sensor
and LED light source with rod dispersion have all been improved for enhanced
image quality with faster scanning speeds. These premium features give you
the leading edge in professional desktop imaging.


1. Multiple film formats (120/220, 35mm, etc.)
2. 4,000 dpi true optical resolution
3. 16-bit A/D converter
4. Large-diameter new Scanner Nikkor ED lens
5. Improved rod dispersion LED illumination
6. High-speed scanning (35mm slide film: 40 seconds; 6 x 9: 185 seconds)
7. Newly-developed, high-quality 3-line CCD sensor
8. New advanced image processing algorithm for colour negative film
9. Multi-sample scanning
10. Quick AF & Quick Preview
11. High-speed IEEE 1394 interface
12. Scan Image Enhancer
13. Digital ICE4 Advanced (TM) Digital ICE Quad Advanced) with Digital ICE
Professional(TM)

Regards
Keith

----------------------------------------------------------
Dr Keith J Morris
Imaging Facilities Manager
Cell Biology Division
Institute of Ophthalmology
University College London
11-43 Bath Street
London EC1V 9EL

Tel: 020 7608 4050
Fax: 020 7608 4034
email: keith.morris-at-ucl.ac.uk


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From: Edward.Calomeni-at-osumc.edu
Date: Wed, 23 Nov 2005 07:52:09 -0600
Subject: [Microscopy] viaWWW: Identifying motor endplates in thick sections

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

Dear all

can I just remind you that Microtek does produce the i900 which is a
large flatbed with full frame glassless holder much as the Agfa duoscan
systems were made. It has a true optical scan resolution of
3200x6400dpi (as far as I know - must be careful here because I know
there was a problem with the early Epsons), 4.2 dynamic range, scans up
to 8x10 inch transpareny or legal format for paper scans, its 48bit
colour USB 2.0 and firewire. It also comes with 'digital ice' and
Silverfast software. Check out the spec on:

http://www.microtek.nl/Product.php?
product=Detail&P_Id=107&Kword=i900&Select=All&FirstData=0

The only thing is that you would have to sort out your own e.m film
holder if it's not one of the following:
SnapTrans holders 35 mm slide holder; 35 mm filmstrip holder; 4"x5"
film holder; 6 x 9 cm film holder. I think it would be the same problem
for any/most of the other scanners. Ironically the do provide a glass
holder as well.

The current UK prices seem to be between about 700 uk pounds which
isn't bad for an 8x10inch glassless scanner - so you could be
talking 'gift-horse' and oral observation.

I don't actually have one but I have used the older Microtek scanmaker
8700 for a couple of years now without problem.

Malcolm

Malcolm Haswell
e.m. unit
School of Health, Natural and Social Sciences
Fleming Building
University of Sunderland
Sunderland
SR1 3SD
UK

e-mail: malcolm.haswell-at-sunderland.ac.uk




----- Original Message -----
X-from: keith.morris-at-ucl.ac.uk

Pat,

Not an easy task to accomplish. Back in the sixty's, the way to ID motor end
plates was to inject methylene blue into the muscle near the nerve while
still in the patient. A book "The Innervation of Muscle: A Biopsy Study" by
C. Coers and A.L. Woolf, published by Charles C. Thomas explains in detail
how this procedure is done.

If specimen is in blocks already, you might try doing immunohistochemistry on
the semi-thin sections using something like S-100 (or another nerve
antibody). Another idea may be to try enzyme histochemistry - with a PAP
secondary step - on the fixed tissue before embedding, using something like
cholinesterase.

I have never tried these, just some random thoughts.

Best of luck,

Ed




Edward P. Calomeni
Director EM Lab
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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Sent: Tuesday, November 22, 2005 9:38 PM
To: Edward Calomeni

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Email: pekysar-at-ucdavis.edu
Name: Pat Kysar

Organization: University of California, Davis, School of Medicine,Pathology

Title-Subject: [Filtered] Identifying motor endplates in thick sections

Question: Hi all,
We have a pathologist who is conducting a study which necessitates the need
to locate motor endplates in resin embedded muscle tissue. He would like to
sucessfully locate them in thick sections so we can accurately cut thin
sections which includes the area of interest.
Does anyone have a protocol for staining the endplates on the thick sections
(epoxy resin embedded) to facilitate a more accurate identification of these
structures? The pathologist has tried finding the areas on Methylene
blue/Azure B stained sections with a very low rate of success and desires a
more accurate method.
Any help would be appreciated.
Thanks,

Pat Kysar
EM Lab
University of California, Davis
School of Medicine, Pathology
1 Shields
Davis, CA 95616
pekysar-at-ucdavis.edu

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From: aolins-at-bowdoin.edu
Date: Wed, 23 Nov 2005 08:16:04 -0600
Subject: [Microscopy] viaWWW: digitizing negatives

Contents Retrieved from Microscopy Listserver Archives
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Email: aolins-at-bowdoin.edu
Name: Ada L. Olins

Organization: Bowdoin College

Title-Subject: [Filtered] digitize a negative

Question:

What are the most critical criteria to consider in order to optimize the quality of a
digitized em negative? Please recommend a scanner or features of a scanner.

I've searched the Listserver archives and not found an answer to my question.

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From: nathano72-at-gmail.com
Date: Wed, 23 Nov 2005 08:16:50 -0600
Subject: [Microscopy] viaWWW: centrifuge tubes?

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Email: nathano72-at-gmail.com
Name: Nathan O'Connor

Organization: Weill Medical College of Cornell University

Title-Subject: [Filtered] centrifuge tubes?

Question: Hi Everyone,

Don't know if such a thing exists (I didn't find anything via a quick search of a few vendors and google), but I'm trying to find centrifuge tubes that auto-open or vent during a spin. Any ideas?

Best regards,
Nathan

--
Nathan O'Connor
Blanchard Lab
Department of Physiology and Biophysics
Weill Medical College of Cornell University
1300 York Avenue
New York, NY 10021


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From: hyi-at-emory.edu
Date: Wed, 23 Nov 2005 08:21:31 -0600
Subject: [Microscopy] Re: TEM negative scanner

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If you all remember, I posted a similar message asking suggestions on
film scanners not long ago. I then did an extensive search myself, and
here is the result.

  First of all, I did not find too many professional grade film
scanners around that will do large format of standard TEM films. Imacon
models were the only ones I found that allows TEM film format, but they
cost over $12K. The first scanner I tried was Nikon SuperCool Scan
900. It has an optical resolution of 4000 dpi, and Dmax of 4.8. It is
perhaps one of the best film scanners for its price (around $2000), but
unfortunately the largest film format it will do is 6 X 8 cm. I looked
into whether or not the film carrier could be modified, but came to a
conclusion that if I really wanted to get this Nikon scanner, I would
have to trim my films, which I do not want to. I then bought an Epson
Perfection 4990 Photo. It is a flatbed about $450. The optical
resolution of this scanner is 4800 dpi, and Dmax 4.0. I consulted with
a few knowledgeable people, and they all think with specifications like
this, it should do an adequate job. But I did not like the images I got
from it at all. It lacks continuous gray tone, so images look contrast
and harsh. 

  The scanner I eventually purchased (about $1000) was Microtek
Artixscan 1800f. It is a dual media unit. The top glass is for
reflective scanning only, but it has a media insert holder that allow
two 4x5" negatives. I can mask off this holder to fit TEM negatives
in. This is better than using a flat bed with transparency adapter
since it eliminates Moire bands (Thank you, Gary Gaugler, for telling
me about this). This scanner offers the highest Dmax than any other
flatbed scanners I could find (Dmax 4.8). But the down side is its
limited resolution (1800 dpi). I debated for a while and decided it
will suit what we need. We usually view our images on a monitor, and
only print in small sizes for publication. We are now using this
scanner with our new Apple 20-inch flat panel cinema, and very happy
with it. However, I think another Microtek scanner, ScanMaker i900
might be a good choice too if not a better one. It cost $600. Rez, 3200
dpi; Dmax, 4.2. 

One thing I learned was that you cannot judge a scanner by its
specifications alone. You really need to try it out to see the end
result. Unfortunately the companies do not set up demos for scanners at
above price range. I end up returned twice and I am sure our purchasing
department was very happy with me. ;-)

Thank you, everyone, and to US readers, enjoy the holiday weekend.

Hong
Emory EM


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From: Elliott-at-arizona.edu
Date: Wed, 23 Nov 2005 11:10:21 -0600
Subject: [Microscopy] printers

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Hi listers
We have been discussing scanners. Thank you all for the input.

What are the current recommendations for printers? There are two
outputs I am interested in (one printer would be good, two is OK)
1) color micrographs
2) TEM micrographs

What do people use? What are you happy with?

Thank you

David


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From: gwe-at-ufl.edu
Date: Wed, 23 Nov 2005 12:26:33 -0600
Subject: [Microscopy] Insect EM

Contents Retrieved from Microscopy Listserver Archives
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Some time ago there was an interesting post describing a method that
allow the cuticle of an insect to bind to the resin so that they did not
separate during sectioning. I printed it out and filed it in a safe
place. now I cannot find it and clearly someone has stolen it. Does
anyone out there recall this post? The person treated the insect with
some stuff during embedding. Willing to pay big for this information.

Greg


-
Gregory W. Erdos, Ph.D,

Assistant Director, Biotechnology Program
Scientific Director, EM Core Lab
P.O. Box 118525
University of Florida
Gainesville, FL 32611
gwe-at-ufl.edu
Phone: 352-392-1295
Fax: 352-846-0251


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From: W.Muss-at-salk.at
Date: Thu, 28 Apr 2005 09:01:02 -0500
Subject: [Microscopy] viaWWW: softening insect cuticle for histological

Contents Retrieved from Microscopy Listserver Archives
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Here you go:

best wishes,
Wolfgang Muss, Salzburg
X-from ListServer's Archive
---------------------------------------------------
X-from: Tindall, Randy D. : TindallR-at-missouri.edu

Some time ago there was an interesting post describing a method that
allow the cuticle of an insect to bind to the resin so that they did not
separate during sectioning. I printed it out and filed it in a safe
place. now I cannot find it and clearly someone has stolen it. Does
anyone out there recall this post? The person treated the insect with
some stuff during embedding. Willing to pay big for this information.

Greg


-
Gregory W. Erdos, Ph.D,

Assistant Director, Biotechnology Program
Scientific Director, EM Core Lab
P.O. Box 118525
University of Florida
Gainesville, FL 32611
gwe-at-ufl.edu
Phone: 352-392-1295
Fax: 352-846-0251


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From: chad.kritzberger-at-yale.edu
Date: Wed, 23 Nov 2005 13:11:29 -0600
Subject: [Microscopy] TEM - Problems with Trichoplax Fixation

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Hello.

I am attemping to fix the small marine invertebrate Trichoplax adhaerens
for conventional TEM usage. My lab grows Trichoplax in large glass
petri dishes where they multiply by asexual budding and binary fission.
Prior to fixation, we transfer the animals to small glass wells with
filtered 36ppt seawater, where they move and lie flat on the bottom of
the glass wells after about 10-20 minutes.

So far, every fixative I've tried results in the animals displaying what
I call a "contraction response." About 3-5 seconds after rapid
immersion in a fixative, the large flat Trichoplax immediately contract
in an all-or-nothing response. They pull themselves inward, develop
frilled edges along their perimeter, and lift off the bottom of the
glass well temporarily before lying back down. Often, the animals curl
in on themselves and are fixed in that position. The degree of this
contraction response is variable, depending on how dissimilar the
fixative is to their normal seawater. For example, a very hypertonic
seawater solution (non-fixative) results in a severe contraction
response, as does seawater buffered in cacodylate. Most of the
fixatives I've tried are 4% glutaraldehyde based. My most successful
fixative is 4% glutaraldehyde in 36ppt seawater, no cacodylate, pH 8.3.
Nevertheless, this fixative causes the Trichoplax to undergo a mild
contraction response. Even introducing a fixative one drop at a time
results in the response.

In short, Trichoplax seems to be very sensitive to any changes in
external environment. The contraction is problematic for my TEM study,
so I am hoping to find a fixative that the animals do not respond to.
Do you have any suggestions regarding either the fixative solution or
the fixative protocol?

Thank you all in advance for your help - it is greatly appreciated!

Chad Kritzberger
chad.kritzberger-at-yale.edu
203-436-1538
Osborne Memorial Labs, Yale University

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From: allan.mitchell-at-stonebow.otago.ac.nz
Date: Wed, 23 Nov 2005 13:27:22 -0600
Subject: [Microscopy] Re: viaWWW: Identifying motor endplates in thick sections

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

Hi Pat

I don't know of a motor end plate stain that can be used on a resin
semi thin section. We would normally locate NMJ by doing a
cytochemical localisation before the processing and embedding.
However, in an 'unstained' muscle block I would look firstly for a
nerve bundle, then for a single nerve fibre running close to the muscle
bundle. If you can locate a single nerve fibre then there is a good
chance a NMJ is not far away.

You do have to develop a bit ofd an eye for them in the semi-thin
section. They are quite small in relation to the other structures and
even at high magn, can be very difficult to see. We describe them to
people looking for one along the edge of a muscle bundle as "look for a
section of the bundle edge that looks a little mottled, perhaps moth
eaten, compared to the rest of the edge. As I say, the single nerve
fibre is a good clue.

Good luck

Allan


On 23/11/2005, at 3:30 PM, pekysar-at-ucdavis.edu wrote:

}
}
}
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} Email: pekysar-at-ucdavis.edu
} Name: Pat Kysar
}
} Organization: University of California, Davis, School of
} Medicine,Pathology
}
} Title-Subject: [Filtered] Identifying motor endplates in thick
} sections
}
} Question: Hi all,
} We have a pathologist who is conducting a study which necessitates the
} need to locate motor endplates in resin embedded muscle tissue. He
} would like to sucessfully locate them in thick sections so we can
} accurately cut thin sections which includes the area of interest.
} Does anyone have a protocol for staining the endplates on the thick
} sections (epoxy resin embedded) to facilitate a more accurate
} identification of these structures? The pathologist has tried finding
} the areas on Methylene blue/Azure B stained sections with a very low
} rate of success and desires a more accurate method.
} Any help would be appreciated.
} Thanks,
}
} Pat Kysar
} EM Lab
} University of California, Davis
} School of Medicine, Pathology
} 1 Shields
} Davis, CA 95616
} pekysar-at-ucdavis.edu
}
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Allan Mitchell
Otago Centre for Electron Microscopy
Department of Anatomy and Structural Biology
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/
Department: http://anatomy.otago.ac.nz/


==============================Original Headers==============================
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From: lgarvie-at-asu.edu
Date: Wed, 23 Nov 2005 15:39:13 -0600
Subject: [Microscopy] SEM - amorphous C staining

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

Chad,

Have you tried cooling the critters first? In the 'frig at 4 deg C. This relaxes or slows many marine inverts so that they don't contract.
Also, MgCl2 is used as an anaesthetic. Try 0.37 M MgCl2.6H20, that should be isotonic to seawater (adjust as needed). This will take a few to maybe 15 minutes.
Or, try a few crystals of MgSO4. Relaxation is slower than with MgCl2, and more may need to be added after 1/2 hour or so.
There are also various funs chemicals to try, most of them narcotics. MS-222 (in the Sigma catalog) works for fish and small crustaceans, it may work on Trichoplax.
Gentle warming, CO2 (as seltzer water or bubbled in), and ethanol added dropwise also work for some animals.

Phil

Philip Oshel
Microscopy Facility Supervisor
Department of Biology
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576
Sic hoc legare scis nimium eruditionis habes.



-----Original Message-----
X-from: chad.kritzberger-at-yale.edu [mailto:chad.kritzberger-at-yale.edu]
Sent: Wed 05/11/23 14:15
To: Oshel, Philip Eugene

Hi all microscopists,

I need advice on making amorphous C visible in the SEM. Basically, I
have an uncovered thin section of a meteorite. This meteorite contains
many small 100 to 1000 nm amorphous C spheres. The C is composed of
small condensed aromatic units that are cross-linked with short
aliphatic chains. I would like to post stain the C so that I can
readily map and find the particles using a backscattered detector on
one of our FEG-SEMs.

I have no experience with sample staining - so of all the stains out
there, which one would be best for my sample?

Thanks,

Laurence



------------------------------------------------------------------------
--------------------------
Dr. Laurence A.J. Garvie
Faculty Research Associate
Department of Geological Sciences
Arizona State University
Tempe
AZ 85287-1404
USA

phone +480 965 0470
fax +480 965 8102
------------------------------------------------------------------------
---------------------------


==============================Original Headers==============================
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From: Colin.Veitch-at-csiro.au
Date: Wed, 23 Nov 2005 19:26:31 -0600
Subject: [Microscopy] Quantomix sample holders

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

Hi,

I noticed that JEOL are selling a product called the Quantomix capsule
which is a specimen holder which allows for the imaging of "wet" samples
in an SEM. I'd be interested in any reports on these as it is something
we are considering.

Cheers


Colin Veitch

Electron Microscopist

CSIRO Textile and Fibre Technology

PO Box 21, BELMONT, Vic. 3216. Australia.

E-mail: colin.veitch-at-csiro.au

Web: http://www.tft.csiro.au

Tel: +61 (0) 3 5246 4000
Mob: 0438 538 475
Fax: +61 (0) 3 5246 4811



The information contained in this e-mail message may be privileged or
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From: jfactor-at-ns.purchase.edu
Date: Wed, 23 Nov 2005 21:22:12 -0600
Subject: [Microscopy] Re: printers

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

I just replaced an older Epson 820, which did a nice job of TEMs, with
nice neutral blacks when set on "black only", but with a somewhat thin
tonal range. HP seems to offer an advantage over Epson (the older ones,
at least), as you get a new print head each time you replace the
cartridge. The old Epson 820 (which has a print head build into the
printer, not in the cartridge), seemed very finicky to me, and needed
constant print head cleaning after a while. However, the older HPs made
grays by using color inks in addition to black, and always seemed to
leave a blue or purple tinge on the grayscale images.

The replacements are two similar, new HP printers, and just today I ran
some test prints of transmission electron micrographs. One 8-1/2x11
printer, the HP 8450, and a larger format printer, the HP 8750; the 8450
is moderately priced, but the 8750 is a bit pricey and seems to offer no
advantage besides the larger format for display prints. Both take three
cartridges (a typical tri-color cartridge, a photo cartridge [stay away
from the "photo blue" which is intended for intense blue skies and gives
TEMs a blue tinge], and a photo gray). It's the photo gray that gives
grayscale images a better tonal range, as it contains black (or dark
gray, I can't be sure which), middle gray, and light gray inks. When set
to "grayscale", with "black cartridge only" (the photo gray cartridge),
the HPs do a very nice job of it, both on Kodak Bright White paper for
drafts, and on high-gloss paper for final prints, and appear to give
very neutral grays. So far, so good (but it's only the first day).

--Jan Factor

---------------------------------------11/23/05
Jan Robert Factor, Ph.D.
Professor of Biology
---------------------------------------
Natural Sciences
Purchase College
State University of New York
735 Anderson Hill Rd.
Purchase, NY 10577
USA
---------------------------------------
Office Tel: 914-251-6659
Office Fax: 914-251-6635
E-mail: jfactor-at-ns.purchase.edu
or- jan.factor-at-purchase.edu
---------------------------------------


Elliott-at-arizona.edu wrote:11/23/05
} ----------------------------------------------------------------------------
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} Hi listers
} We have been discussing scanners. Thank you all for the input.
}
} What are the current recommendations for printers? There are two
} outputs I am interested in (one printer would be good, two is OK)
} 1) color micrographs
} 2) TEM micrographs
}
} What do people use? What are you happy with?
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} David
}
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From: David.Patton-at-uwe.ac.uk
Date: Thu, 24 Nov 2005 05:44:47 -0600
Subject: [Microscopy] TEM - Problems with Trichoplax Fixation

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

No experience but.... I have read that osmium tetroxide can be used as a
primary fixative for marine invertebrates It might work faster as well.
Take care to use a sealed container (it is very volatile) if trying to
observe outside of a fume cupboard.

Dave

-----Original Message-----
X-from: chad.kritzberger-at-yale.edu [mailto:chad.kritzberger-at-yale.edu]
Sent: 23 November 2005 19:16
To: David Patton

Hello.

I am attemping to fix the small marine invertebrate Trichoplax adhaerens
for conventional TEM usage. My lab grows Trichoplax in large glass
petri dishes where they multiply by asexual budding and binary fission.
Prior to fixation, we transfer the animals to small glass wells with
filtered 36ppt seawater, where they move and lie flat on the bottom of
the glass wells after about 10-20 minutes.

So far, every fixative I've tried results in the animals displaying what
I call a "contraction response." About 3-5 seconds after rapid
immersion in a fixative, the large flat Trichoplax immediately contract
in an all-or-nothing response. They pull themselves inward, develop
frilled edges along their perimeter, and lift off the bottom of the
glass well temporarily before lying back down. Often, the animals curl
in on themselves and are fixed in that position. The degree of this
contraction response is variable, depending on how dissimilar the
fixative is to their normal seawater. For example, a very hypertonic
seawater solution (non-fixative) results in a severe contraction
response, as does seawater buffered in cacodylate. Most of the
fixatives I've tried are 4% glutaraldehyde based. My most successful
fixative is 4% glutaraldehyde in 36ppt seawater, no cacodylate, pH 8.3.
Nevertheless, this fixative causes the Trichoplax to undergo a mild
contraction response. Even introducing a fixative one drop at a time
results in the response.

In short, Trichoplax seems to be very sensitive to any changes in
external environment. The contraction is problematic for my TEM study,
so I am hoping to find a fixative that the animals do not respond to.
Do you have any suggestions regarding either the fixative solution or
the fixative protocol?

Thank you all in advance for your help - it is greatly appreciated!

Chad Kritzberger
chad.kritzberger-at-yale.edu
203-436-1538
Osborne Memorial Labs, Yale University

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18, 33 -- From: David Patton {David.Patton-at-uwe.ac.uk}
18, 33 -- Subject: RE: [Microscopy] TEM - Problems with Trichoplax Fixation
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From: hinmeigeng-at-hotmail.com
Date: Thu, 24 Nov 2005 06:32:50 -0600
Subject: [Microscopy] Contrast and focus in TEM

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

Hi All!

I've just been looking at some thin crystals of an organic compound, sitting
on a carbon film, under bright-field TEM. When in exact focus, they appear
much less constrasty than when the focus knob is turned to the left of the
right. When turned to the right, dark fringes appear around the crystals.
I would like to ask:

(1) Which of the two , left or right, is over/under focus?

(2) Why are they more constrasty overall when over or under-focussed?

(3) Why are they more contrasty at lower magnification?

(Things were much easier when looking at shadowed-carbon replicas of polymer
surfaces!)

Any help would be much appreciated.

-----------------------------------
Robert H. Olley
Reply to: R.H.Olley-at-reading.ac.uk
URL: http://www.rdg.ac.uk/~spsolley
-----------------------------------



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From: jb_sanderson-at-yahoo.com
Date: Thu, 24 Nov 2005 07:35:56 -0600
Subject: [Microscopy] 3 open positions for miscoscopists at the Max Planck Institute (MPI-CBG), Dresden, Germany

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

Dear all,

I would like to ask you for your help:

The Max Planck Institute of Molecular Cell Biology
and Genetics in Dresden, Germany is now advertising
for good people who would be willing to help us in
running core facilities in our international
institute ( more than 150 scientists from more then 30
countries). Currently there are three positions open:

Senior Microscopist (Imaging Specialist) - Light
Microscopy Facility
http://www.mpi-cbg.de/research/jobs/lmf.html

Facility Leader - Image Analysis Facility

http://www.mpi-cbg.de/research/jobs/imageanalysis.html

Facility Leader - Electron Microscopy Facility
http://www.mpi-cbg.de/research/jobs/em.html

If you are interested in these positions, please do
not hesitate to contact me.

If you know someone who might be also interested in
one of the above mentioned positions, please forward
my email to him/her. The deadline for the appliactions
is 9. December 2005.
Thank you very much for your kind assistance in
advance.

Regards

Jan

xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
Jan Peychl, M.D., Ph.D.
Service Leader
Light Microscopy Facility
Max Planck Institute of Molecular Cell Biology and
Genetics
Pfotenhauerstrasse 108
01307 Dresden
Germany

Tel.: +49 351 210 2502
Fax: +49 351 210 2000
web: www.mpi-cbg.de






___________________________________________________________
Yahoo! Messenger - NEW crystal clear PC to PC calling worldwide with voicemail http://uk.messenger.yahoo.com

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From: Philip.Koeck-at-biosci.ki.se
Date: Fri, 25 Nov 2005 02:27:08 -0600
Subject: [Microscopy] Contrast and focus in TEM

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

Robert

I'm sure you will get much more detailed answers when the USA wakes up,
but here's my two pence worth.

When you turn to the right and see a dark fringe at the edge of objects
then it is overfocussed and a light fringe is underfocussed when you
turn to the left. These are Fresnel fringes and are produced because of
diffraction contrast which is a result
of scattering of light/electrons at edges. It can and is quite often
acceptable to defocus slightly to increase contrast but this should
normally only be slightly underfocussed. The logic being that bright
fringes will enhace the dark edges beside them, but dark fringes will
introduce more apparent dark structures.

When you examine specimens at low magnification amplitude contrast
makes a greater contribution to the image than phase contrast.
Amplitude contrast is generated by the
influence of the specimens atomic nuclei on electrons in the beam. The
higher the mass of the nucleus the greater the 'scattering power' of
that area of the specimen and the more electrons scattered out of the
main beam path the darker that area. Amorphous samples simply produce
more or less contrast by mass alone although crystals scatter electrons
in much more defined ways due to the interaction of the 'wavelength' of
the electrons, the lattice spacing in the crystal and its angle of tilt.

There are many
good books on this subject, but here's the first one that came to hand:
Principles and Practice of Electron Microscope Operation; A.W. Agar,
R.H. Alderson and D. Chescoe; Pub North Holland (3rd print 1980) ISBN 0
72044255 9 where chapter 3 on image formation is particularly
appropriate.

My apologies to crystallographers everywhere for a simple biologists
explanation of diffraction contrast without once mentioning Bragg.

Malcolm

Malcolm Haswell
e.m. unit
School of Health, Natural and Social Sciences
Fleming Building
University of Sunderland
Sunderland
SR1 3SD
UK
tel no: +44 (0)191 515 2872
e-mail: malcolm.haswell-at-sunderland.ac.uk


----- Original Message -----
X-from: hinmeigeng-at-hotmail.com

Your description is right on target. When I was learning to use the
em (in the dark ages, with an RCA-EMU3), I was taught to obtain best
focus by experimenting first with a holey sample so that I could
adjust what my eyes thought was best focus (because we are uniquely
sensitive to contrast) to the true focus. I then would focus on a
sample to what I thought was best, and then adjust the focus setting
by a few notches. The issue of whether you want to include the
underfocus fringes in your image becomes, then, an issue of whether
you want to include the optical artifact for contrast, or you are
interested in the "true" edge of your objects.


Date sent: Thu, 24 Nov 2005 08:14:13 -0600
To: jbs-at-temple.edu
X-from: malcolm.haswell-at-sunderland.ac.uk
Send reply to: malcolm.haswell-at-sunderland.ac.uk

Hello,

I might be wrong, but seem to have a little different understanding from
the posts so far to these concepts of very importance in both theory and
practice so that I reply, hoping to solicit more insights. In short,

1) Slight under-focus (OL knob counterclockwise adjustment from the
least contrast or true focus point) not only makes image look good, but
provides better resolution.

2) There are only two types of contrast mechanisms in my opinion:
amplitude contrast and phase contrast. Amplitude contrast includes a)
diffraction contrast, b) mass-thickness contrast, and c) Z-contrast (a
special case of mass-thickness contrast).

Also due to the locality nature, Z-contrast and phase contrast can
normally be thought responsible for or capable of providing lattice
resolution (you may argue with in the cases of imaging with several
diffraction spots) or atomic resolution.

Happy holiday or vacation (to those who don't view Thanksgiving as a
holiday)!

Chaoying Ni
W.M. Keck Electron Microscopy Facility
Facility location: 022 Spencer Laboratory
Mailing address:
201 duPont Hall
Dept. of Materials Sci. and Eng.
University of Delaware
Newark, DE 19716
(302) 831-2318 (Phone) (302) 831-4545 (Fax)
http://eml.masc.udel.edu




-----Original Message-----
X-from: malcolm.haswell-at-sunderland.ac.uk
[mailto:malcolm.haswell-at-sunderland.ac.uk]
Sent: Thursday, November 24, 2005 9:16 AM
To: cni-at-UDel.Edu

Robert

I'm sure you will get much more detailed answers when the USA wakes up,
but here's my two pence worth.

When you turn to the right and see a dark fringe at the edge of objects
then it is overfocussed and a light fringe is underfocussed when you
turn to the left. These are Fresnel fringes and are produced because of
diffraction contrast which is a result
of scattering of light/electrons at edges. It can and is quite often
acceptable to defocus slightly to increase contrast but this should
normally only be slightly underfocussed. The logic being that bright
fringes will enhace the dark edges beside them, but dark fringes will
introduce more apparent dark structures.

When you examine specimens at low magnification amplitude contrast
makes a greater contribution to the image than phase contrast.
Amplitude contrast is generated by the
influence of the specimens atomic nuclei on electrons in the beam. The
higher the mass of the nucleus the greater the 'scattering power' of
that area of the specimen and the more electrons scattered out of the
main beam path the darker that area. Amorphous samples simply produce
more or less contrast by mass alone although crystals scatter electrons
in much more defined ways due to the interaction of the 'wavelength' of
the electrons, the lattice spacing in the crystal and its angle of tilt.

There are many
good books on this subject, but here's the first one that came to hand:
Principles and Practice of Electron Microscope Operation; A.W. Agar,
R.H. Alderson and D. Chescoe; Pub North Holland (3rd print 1980) ISBN 0
72044255 9 where chapter 3 on image formation is particularly
appropriate.

My apologies to crystallographers everywhere for a simple biologists
explanation of diffraction contrast without once mentioning Bragg.

Malcolm

Malcolm Haswell
e.m. unit
School of Health, Natural and Social Sciences
Fleming Building
University of Sunderland
Sunderland
SR1 3SD
UK
tel no: +44 (0)191 515 2872
e-mail: malcolm.haswell-at-sunderland.ac.uk


----- Original Message -----
X-from: hinmeigeng-at-hotmail.com

Hi,

It seems about time to add a post from California (yes, some of us are
finally awake on the left coast).

As far as I can see, both (all) of the contributed points of view are
correct.

Underfocus by a few hundred Angstroms will indeed enhance contrast and
improve resolution. The explanation was first given by Otto Scherzer in
his famous "20/20" paper on page 20 of volume 20 of J. Appl. Phys.
[Scherzer, O. (1949). "The theoretical resolution limit of the electron
microscope" J. Appl. Phys. 20, 20-29].

Scherzer describes how an underfocus of minus sqrt (Cs * wavelength)
optimizes transfer of spatial frequencies into the image from the
electron wave leaving the specimen -- by balancing (positive) phase
shifts due to spherical aberration with (negative) shifts from
underfocus to form a kind of quarter-wave plate. Optimum underfocus
ranges from -750 Angstrom for a spherical aberration of 1 mm at 100 kV
to half that for a Cs of 0.5 mm at 300 kV.

Be aware that too much underfocus (or overfocus) will add too much
negative (or positive) phase shift and "scramble" the image so it no
longer shows a simple projection of the specimen.

There is more on high resolution and focus in many publications -- the
one I like is the paper "Resolution in high-resolution electron
microscopy", M.A. O'Keefe, Ultramicroscopy 47 (1992) 282-297. ;-)

For TEM of organic molecules see papers by John Fryer -- especially J R
Fryer 1993 J. Phys. D: Appl. Phys. 26 B137-B144. There's also M.A.
O'Keefe, J.R. Fryer and D.J. Smith, Acta Cryst. A39 (1983) 838-847.

Happy Thanksgiving,
Mike

Michael A. O'Keefe
National Center for Electron Microscopy
Lawrence Berkeley National Laboratory
Berkeley, CA 94720

----- Original Message -----
X-from: cni-at-udel.edu

p.s. If anyone would like a calculator program (PC only) that will
compute several TEM parameters, such as optimum defocus, let me know and
I'll email it to you. It uses reverse Polish -- to compute optimum
defocus for a voltage of 300kV and Cs of 1.2 mm, you would enter "1.2
300 wavl opt".
Mike
----- Original Message -----
X-from: MAOKeefe-at-lbl.gov

Hi,

You might want to have a look at a few useful webpages:

a focusing simulator:
http://www.umsl.edu/~fraundor/epc/index.html

contrast transfer function:
http://clik.to/ctfexplorer
http://ncmi.bcm.tmc.edu/homs/wen/ctf/ctfapplet.html



As we grow older we acquire wisdom,
but can't remember where we put it.- DA

Philip Koeck
Karolinska Inst. and University College of S. Stockholm
Dept. of Bioscience at Novum
tel: +46 8 608 9186
fax: +46 8 608 9290
web: http://www.csb.ki.se/users/philip/philown.html


-----Original Message-----
X-from: hinmeigeng-at-hotmail.com [mailto:hinmeigeng-at-hotmail.com]
Sent: 24 November 2005 13:37
To: Philip.Koeck-at-biosci.ki.se

Hi All!

I've just been looking at some thin crystals of an organic compound, sitting

on a carbon film, under bright-field TEM. When in exact focus, they appear
much less constrasty than when the focus knob is turned to the left of the
right. When turned to the right, dark fringes appear around the crystals.
I would like to ask:

(1) Which of the two , left or right, is over/under focus?

(2) Why are they more constrasty overall when over or under-focussed?

(3) Why are they more contrasty at lower magnification?

(Things were much easier when looking at shadowed-carbon replicas of polymer

surfaces!)

Any help would be much appreciated.

-----------------------------------
Robert H. Olley
Reply to: R.H.Olley-at-reading.ac.uk
URL: http://www.rdg.ac.uk/~spsolley
-----------------------------------



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From: microfrog9-at-hotmail.com
Date: Fri, 25 Nov 2005 07:46:00 -0600
Subject: [Microscopy] viaWWW: Phase Contrast Microscope - Phase Plate

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Title-Subject: [Filtered] Phase Contrast Microscope - Phase Plate

Question: Hi,

Is there an equation to determine the depth of the etched region of a phase plate ?

Thanks

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From: neapit-at-yahoo.com
Date: Fri, 25 Nov 2005 08:15:49 -0600
Subject: [Microscopy] viaWWW:Microscopy & Molecular biology

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Organization: Medical school University of Athens-Greece

Title-Subject: [Filtered] Molecular biology

Question: Hi everybody.
I'm a PhD student in the med school with a degree in chemistry. I want to know where I can found info about microscopy techniques in the field of molecular biology.

thank you for helping me
Nea

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From: weis183-at-yahoo.fr
Date: Fri, 25 Nov 2005 12:46:21 -0600
Subject: [Microscopy] TEM calibration standard

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

Hi all,

I'm looking for a reference sample for calibration
of camera lenght for diffraction and magnification
(mainly for High Resolution).
I already tried several sample (sold by Agar
scientific):

"Oriented single crystal gold foil" for camera
length and high resolution
"Evaporated aluminium film" for camera length
"Evaporated Thallous chloride" for camera length and
High Resolution
"Graphitised Carbon Black" for camera length and
High Resolution.

However I obtain some non negligible shift on
dspacings between various calibration standards
(around 1% or 2%) these shifts are reproducible and
thus are not due to alignement problem or bad
eucentric position.

As this sample are not sold with a control
certifical does someone know their real accuracy? Or
is their an explanation to this errors?

A more expensive calibration standard "MAG*I*CAL" is
sold, does someone has experienced it?

Any help would be much appreciated.

Patrick Weisbecker
LCTS / PESSAC
FRANCE







___________________________________________________________________________
Appel audio GRATUIT partout dans le monde avec le nouveau Yahoo! Messenger
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From: tom-at-tomkaye.com
Date: Sat, 26 Nov 2005 22:45:32 -0600
Subject: [Microscopy] Robinson Detector Repair??

Contents Retrieved from Microscopy Listserver Archives
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I would like to start off with two questions.
Is there any way that you might have prepared this sample without embedding and preparing it as a thin section? How are you sure that such C spheres are present?
 
I expect that the C spheres would virtually disappear in the epoxy embedding medium which prompts your question about staining. If no epoxy was present, I suspect the spheres would show up nicely against the light background of the meteorite.
 
Since you cannot very well be using BSE to image the sample, how do you know the spheres are present and how do you know their chemical nature or size? If someone else has suggested or claimed their presence, how did they perform the characterization?
 
I would be interested in hearing the outcome of this exercise. It sounds like a challenge to do this by BSE even in a FEG-SEM, but I could be mistaken.
 
Warren Straszheim
Iowa State University

________________________________________
X-from: lgarvie-at-asu.edu [mailto:lgarvie-at-asu.edu]
Sent: Wed 11/23/2005 3:40 PM
To: wesaia-at-iastate.edu

Hello All,

I have a Robinson Detector that is problematic and in need of repair.
Strangely enough, a search of the web comes up blank on the manufacturer or
places that fix them.

Does anyone have contact info?

Great list by the way! As the rare hobbyist with an SEM, this is my only
contact with the EM world and it's much appreciated.

Tom Kaye




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From: cgarber-at-2spi.com
Date: Sun, 27 Nov 2005 08:35:40 -0600
Subject: [Microscopy] Robinson BSE detectors website

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

Tom Kaye wrote:
======================================================
Hello All,

I have a Robinson Detector that is problematic and in need of repair.
Strangely enough, a search of the web comes up blank on the manufacturer or
places that fix them.

Does anyone have contact info?

Great list by the way! As the rare hobbyist with an SEM, this is my only
contact with the EM world and it's much appreciated.
=====================================================
The website you are looking for is
http://www.etpsemra.com.au/

Dr. Vivian Robinson runs the company "hands on" and if you contact them
through their website in Australia, you could very well hear from Dr.
Robinson himself.

SPI Supplies has been providing sales and service for their detectors, see
URL
http://www.2spi.com/catalog/instruments/robinson-backscattered-electron-BSE-detector.shtml

Alternatively, you can contact me off-line and I will try my best to give
you the assistance you require.

Chuck
============================================

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

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







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From: cgarber-at-2spi.com
Date: Sun, 27 Nov 2005 12:11:40 -0600
Subject: [Microscopy] =?Windows-1252?Q?Quantomix=AE_sample_holders?=

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

Colin Veitch wrote:
========================================================
I noticed that JEOL are selling a product called the Quantomix capsule
which is a specimen holder which allows for the imaging of "wet" samples
in an SEM. I'd be interested in any reports on these as it is something
we are considering.
========================================================
The Quantomix® capsules certainly do "work". I have seen them demonstrated
on several different occasions at trade shows and on the SEMs of several
different manufacturers. The technique is very exciting.

However I mention there is an alternative way to make an environmental (
vacuum compatible) cell, and that is with the use of silicon nitride
membrane window grids, two glued together, see URL
http://www.2spi.com/catalog/instruments/silicon-nitride.shtml

There is an example of the use of the SPI Supplies® Brand silicon nitride
membrane window grids toward the bottom of the page for an "environmental
cell" application.

Disclaimer: SPI Supplies offers a competing system to the Quantomix
capsules so we would have a vested interest in promoting the use of our SPI
Supplies® Brand silicon nitride membrane window grid system instead of the
polymeric window Quantomix capsules.

Chuck

============================================

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

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







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From: schooley-at-mcn.org
Date: Sun, 27 Nov 2005 13:56:52 -0600
Subject: [Microscopy] children's microscopy

Contents Retrieved from Microscopy Listserver Archives
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It's holiday shopping time again. Good books about magnification
written for the beginning reader are scarce. If your young one will
be getting a dissecting scope or a magnifying glass this year, there
are two new small, inexpensive pamphlets that you should consider:
"Looking Through a Microscope" & "You can Use a Magnifying Glass".
You'll find information on both in the MICRO bibliography (URL
below); enter "supplimental books", "for the primary grades", &
"recommended" in the search engine.
--
Caroline Schooley
Project MICRO Coordinator
Microscopy Society of America
Box 117, 45301 Caspar Point Road
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.microscopy.org/ProjectMICRO

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From: pgrover-at-bilbo.bio.purdue.edu
Date: Mon, 28 Nov 2005 08:20:34 -0600
Subject: [Microscopy] $ 1 stocking stuffers

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

Surplus Shed (surplusshed.com) has folding doublet 10X magnifiers for $1.25
ea. (or 5/5.00). Glass lenses, aluminum & plastic housing. I like to drill
a hole through plastic hinge area to attach a neck string. Great stocking
stuffers, party favors, souvenirs of a field trip to your lab, or just carry
a few around in your pocket to give to youngsters or leave them in strategic
locations for someone to find (random act of kindness?). Price of a candy
bar. I'm ordering my second 100 of them. I reckon if it gets one kid in 10
away from the video game or TV for a little while, it's money well spent.

Disclaimer: I have no connection to Surplus Shed; I'm just cheap, and like
to pass along a 'steal' when I see one.

Paul

----------------------------------------------------------------------------
The world is so full of a number of things
I wonder we're not all as happy as kings

--Robert Louis Stevenson




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From: marko-at-wadsworth.org
Date: Mon, 28 Nov 2005 09:12:27 -0600
Subject: [Microscopy] Invitation to organize symposia for M&M2007

Contents Retrieved from Microscopy Listserver Archives
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***Second announcement--confirmations will start end of December***

We are still looking for some great ideas for symposia at M&M2007!

Microscopy & Microanalysis 2007 Meeting

August 6-9, 2007
Broward County Convention Center
Ft. Lauderdale, Florida

Co-sponsored by

The Microscopy Society of America
The Microbeam Analysis Society
The International Metallographic Society

Proposals are coming in. We have suggestions for some of the customary
symposia, but have signed up only four organizers of these. Otherwise, the
program is still largely open at this time.

We would like the majority of the proposals to be submitted by the end of
the year. We will start sending out acceptance letters in late December,
and by mid-February we expect the program to be mostly filled.

This timetable is considerably accelerated in comparison with previous
years, and we now require a description of 150-300 words for each proposed
symposia. The description should take the form of those found in the Call
for Papers and Expo of past years; it should be an announcement of the
symposium and an invitation for contributions. The Program Committee will
select symposia based on these descriptions, so that overlap will be
minimized and symposia will complement each other to form a coherent
overall program.

You need not be a member of MSA, MAS, or IMS to propose a symposium,
although we hope that your experience with the M&M meeting will encourage
you to join.

Please send your suggestion (complete with description) directly to the
Program Chair, or to the M&M2007 Co-Chair of your Society.

The M&M2007 website is

http://mm2007.microscopy.org/

Early next year, symposia will be posted on the website as they are accepted.

Program contacts:

Mike Marko, Program Chair (marko-at-wadsworth.org)
John Henry Scott, Vice Program Chair (johnhenry.scott-at-nist.gov)
Ed Vincenzi, MAS Program Co-Chair (vicenzi-at-volcano.si.edu)
Steve Dekanich, IMS Program Co-Chair (dekanichsj-at-y12.doe.gov)

Local Arrangments contact:

Lucille Giannuzzi, Local Arrangements Chair (lgiannuzzi-at-feico.com)

Meeting Management contact:

Phillip Ridley, Conference Manager
Bostrom Corp
230 East Ohio, Suite 400
Chicago, IL 60611
Tel: 312-644-0828
Fax: 312-644-8557
Email: pridley-at-bostrom.com





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From: morgansbearhunter-at-yahoo.com
Date: Mon, 28 Nov 2005 10:56:38 -0600
Subject: [Microscopy] AskAMicroscopist: tem embedding

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This Question was submitted to Ask-A-Microscopist by (morgansbearhunter-at-yahoo.com)
from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, November 28, 2005 at 10:49:22
Remember to consider the Grade/Age of the student when considering the Question
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Email: morgansbearhunter-at-yahoo.com
Name: Kathryn Privett

Organization: Charlotte Healthcare System

Education: Graduate College

Location: charlotte, NC,

Title: tem embedding

Question: Lately when I embed clinical tissue in blocks I get tiny little holes all over the tissue. I changed the dehydration Etoh, the propylene oxide. Now I am wondering if it could be the spurrs NSA. We ordered it and received in 2002. Do you have any suggestions?
Kathryn Privett

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

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From: milton.charlton-at-utoronto.ca
Date: Mon, 28 Nov 2005 11:25:07 -0600
Subject: [Microscopy] viaWWW: Jena operating scope parts

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using the WWW based Form at http://www.microscopy.com/MicroscopyListserver/MLFormMail.html
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Email: milton.charlton-at-utoronto.ca
Name: Milton Charlton

Organization: University of Toronto

Title-Subject: [Filtered] Jena operating scope parts

Question: Could anyone tell me where to get a replacement rack and pinion foucusing mechanism for a Jena operating scope that was made in the 1960's?
Thanks
M. Charlton
University of Toronto

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

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From: hinmeigeng-at-hotmail.com
Date: Mon, 28 Nov 2005 14:15:30 -0600
Subject: [Microscopy] Thanks (TEM) + Question (SEM)

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Many thanks to y'all who replied to my question about under/over focus in
TEM. I think I have enough of an idea now.

Now for a SEM question. I've seen many questions about grain size of
sputtered gold on this list, but what about the tendency of gold films to
greak up into a sub-micron "crazy-paving" structure? Does anyone know what
causes this, and how to cure it? Often the crazy paving totally dominates
the picture, and one has to mentally filter it out to see the underlying
structure.

-----------------------------------
Robert H. Olley
Reply to: R.H.Olley-at-reading.ac.uk
URL: http://www.rdg.ac.uk/~spsolley
-----------------------------------



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From: bozzola-at-siu.edu
Date: Mon, 28 Nov 2005 14:49:32 -0600
Subject: [Microscopy] Re: Thanks (TEM) + Question (SEM)

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One (prehaps the major) cause of the "crazy-paving" or "dried river
bed" appearance of specimens that were coated with heavy metals for
SEM viewing is the expansion and contraction of the underlying
specimen. Since the metal coating has little flex/stretch
capabilities, it will break (or shatter) if the underlying specimen
flexes or expands. The causes of the expansion of the specimen may
be: absorption/loss of moisture (as one goes in and out of the
vacuum), heating/cooling, mechanical flexing. This can be minimized
by keeping the specimens always in a dry environment, at a standard
temperature and not bending/flexing them.

} Now for a SEM question. I've seen many questions about grain size of
} sputtered gold on this list, but what about the tendency of gold films to
} greak up into a sub-micron "crazy-paving" structure? Does anyone know what
} causes this, and how to cure it? Often the crazy paving totally dominates
} the picture, and one has to mentally filter it out to see the underlying
} structure.
}
} -----------------------------------
} Robert H. Olley
} Reply to: R.H.Olley-at-reading.ac.uk
} URL: http://www.rdg.ac.uk/~spsolley
} -----------------------------------


--
##############################################################
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
750 Communications Drive - MC 4402
Southern Illinois University
Carbondale, IL 62901 U.S.A.
Phone: 618-453-3730
Email: bozzola-at-siu.edu
##############################################################

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From: walck-at-southbaytech.com
Date: Mon, 28 Nov 2005 15:04:37 -0600
Subject: [Microscopy] Thanks (TEM) + Question (SEM)

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The quick answer to your question concerning the structure of gold
coatings is to not use gold coatings. Gold is notorious for nucleating
islands on the surface that grow and eventually coalesce to form the
coating. This growth pattern results in your "crazy-paving" structure.
In fact, the resolution test sample for SEM has traditionally been a
gold coating evaporated onto a smooth carbon substrate and the minimum
distance between adjacent gold islands is used as the acceptance
measurement for the microscope. Au-Pd will offer a better coating on
inexpensive desktop coaters than plain gold, but you will still see the
grain structure at higher magnifications. Ion Sputter coating systems
such as our IBS/e system can sputter numerous different materials. The
materials that offer extremely good, uniform, and small structure
suitable for high resolution SEM work are Pt, Pd, Cr, Ir, and W.

If you look up Elaine Humphrey's response to a similar question on this
listserver from earlier this year, she gives a link to a web site that
shows comparisons of coatings from different materials using one of our
competitors' systems. (Sorry, I hope you understand why I don't give
out the URL.)

-Scott

Disclaimer: South Bay Technology, Inc. manufactures and sells the IBS/e
ion beam sputter deposition and etching system.

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
walck-at-southbaytech.com


-----Original Message-----
X-from: hinmeigeng-at-hotmail.com [mailto:hinmeigeng-at-hotmail.com]
Sent: Monday, November 28, 2005 12:23 PM
To: Walck-at-SouthBayTech.com


Many thanks to y'all who replied to my question about under/over focus
in
TEM. I think I have enough of an idea now.

Now for a SEM question. I've seen many questions about grain size of
sputtered gold on this list, but what about the tendency of gold films
to
greak up into a sub-micron "crazy-paving" structure? Does anyone know
what
causes this, and how to cure it? Often the crazy paving totally
dominates
the picture, and one has to mentally filter it out to see the underlying

structure.

-----------------------------------
Robert H. Olley
Reply to: R.H.Olley-at-reading.ac.uk
URL: http://www.rdg.ac.uk/~spsolley
-----------------------------------



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From: mike.reedy-at-cellbio.duke.edu
Date: Mon, 28 Nov 2005 16:35:40 -0600
Subject: [Microscopy] Re: AskAMicroscopist: tem embedding

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Kathryn
My guess is, the holes will go away if you use a
dehumidifier to bring the relative humidity in
your work-room (or work-chamber) below 60-70%
before, during and after the last change of 100%
accelerated resin. We used to get episodes of
holes in thin sections of our Araldite
506-DDSA-DER 736 resin mixture until we followed
the advice we found in this paper:

(1977) H. D. Dellman and C. Pearson Better
epoxy resin embedding for electron microscopy at
low relative humidity. Stain Technol. 52:5-8.

This is essential for us, perhaps moreso than for
most labs, because we need a dehumidified
environment during the 15-90 minutes it may take
us to lovingly manipulate fully infiltrated
single muscle fibers and rafts of 3-5 single
fibers into position on regions of dry substrate
with only a minimal micro-meniscus of external
resin; once thus positioned, they stick
themselves nicely to each other and to the smooth
substrate of polypropylene sheet (by surface
tension) during the first 2-4 hours of cure at
60° C. We then invert a full BEEM capsule of
liquid resin over the fiber or raft and finish
the cure overnight at 80°C. This gives us
superbly oriented single fibers exactly parallel
to the flat surface of the cured block and lying
within 3-5 microns of the resin surface; we like
to use it to co-embed fibers from different
experiments in a single raft, so we can make one
longitudinal (or cross-) section of a such a
combi-block do the work of 3-5 sections of
separate blocks.

The very high surface-to-volume ratio of this
"dry, flat" embedding procedure makes the resin
very susceptible to humidity during the pre-cure
manipulations. So we go for as low a humidity as
our commercial dehumidifier can achieve in our
small workroom, often below 50% if we run the
thing all night in a closed room. Obviously even
lower levels could be rapidly attained if the
dehumidified outflow from the appliance were
delivered into a small desktop working chamber,
but we've not found that to be necessary for our
work.

The curing oven itself is not a worry-- its
internal RH is below 10%, if i recall our few
measurements some years ago. So we typically
accelerate infiltration itself in 100%
accelerated resin mixture by putting the vials on
a sloped rotator in the 60°C oven, and
remembering to change the resin for fresh every
30 minutes, x2 or x3. In that heat, the resin
mixture becomes so water-thin, and stays that way
for at least 40 minutes, that I've long supposed
even Spurr's or similar could not become
significantly lower in viscosity or penetrate
the tissue any more completely.

-mike reedy-

At 11:04 AM -0600 11/28/05, morgansbearhunter-at-yahoo.com wrote:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America


--
-mike reedy-

************************
Michael K. Reedy, M.D.
Duke Univ. Med. Center
Dept. Cell Biology, Box 3011 (for U.S. Mail)
458 Alex Sands Bldg, Research Dr (courier)
Durham, NC 27710

Office 919-668-2534
Lab 919-684-5674
Fax 919-681-9929
mike.reedy-at-cellbio.duke.edu
http://note.cellbio.duke.edu/Faculty/Research/Reedy.html


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From: tivol-at-caltech.edu
Date: Mon, 28 Nov 2005 17:50:45 -0600
Subject: [Microscopy] Re: TEM calibration standard

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On Nov 25, 2005, at 10:46 AM, weis183-at-yahoo.fr wrote:

} A more expensive calibration standard "MAG*I*CAL" is
} sold, does someone has experienced it?
}
Dear Patrick,
I have used the MAG*I*CAL, and it works very well. You can calibrate
up to the highest mags on your scope using the Si lattice, and you can
calibrate camera lengths. If you have a tilt-rotation stage, you can
orient the MAG*I*CAL precisely to do all the calibrations, and with a
single-tilt stage you will have to orient the grid within the stage for
best results. The only problem with it is that it should be used at
room temperature, which is not a big deal except for cryo facilities.
Yours,
Bill Tivol, PhD
EM Scientist and Manager
Cryo-Electron Microscopy Facility
Broad Center, Mail Code 114-96
California Institute of Technology
Pasadena CA 91125
(626) 395-8833
tivol-at-caltech.edu


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From: bozzola-at-siu.edu
Date: Mon, 28 Nov 2005 19:52:06 -0600
Subject: [Microscopy] Re: TEM calibration standard

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

I concur with Bill Tivol on the MAG*I*CAL calibration standard. We
have been using it since it first came out several years ago and have
been extremely pleased with it as it covers the entire magnification
range of the TEM and can be used to calibrate camera lenth, etc.

You need to be very careful with it (not for students to manipulate)
since it is brittle and easily broken. Yes, I broke one (a $900
mistake) when I put it back onto the membrane of the storage box. It
is best to allow it to drop gently onto the membrane since if you
misjudge the exact location of the membrane you can flex and break it.

Go for it and handle with care.

JB




} On Nov 25, 2005, at 10:46 AM, weis183-at-yahoo.fr wrote:
}
} } A more expensive calibration standard "MAG*I*CAL" is
} } sold, does someone has experienced it?
} }
} Dear Patrick,
} I have used the MAG*I*CAL, and it works very well. You can calibrate
} up to the highest mags on your scope using the Si lattice, and you can
} calibrate camera lengths. If you have a tilt-rotation stage, you can
} orient the MAG*I*CAL precisely to do all the calibrations, and with a
} single-tilt stage you will have to orient the grid within the stage for
} best results. The only problem with it is that it should be used at
} room temperature, which is not a big deal except for cryo facilities.
} Yours,
} Bill Tivol, PhD
} EM Scientist and Manager
} Cryo-Electron Microscopy Facility
} Broad Center, Mail Code 114-96
} California Institute of Technology
} Pasadena CA 91125
} (626) 395-8833
} tivol-at-caltech.edu
}
}
} ==============================Original Headers==============================
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From: cgarber-at-2spi.com
Date: Mon, 28 Nov 2005 23:00:33 -0600
Subject: [Microscopy] Crazy paving structures in gold sputtered films

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

Robert H. Olley wrote:
========================================================
Now for a SEM question. I've seen many questions about grain size of
sputtered gold on this list, but what about the tendency of gold films
to greak up into a sub-micron "crazy-paving" structure? Does anyone know
what causes this, and how to cure it? Often the crazy paving totally
dominates the picture, and one has to mentally filter it out to see the
underlying structure.
========================================================
Our philosophy has always been that if this is happening, then you are
probably putting on too much gold and/or exposing the sample to too much
heat. Other factors can also increase the chances of seeing this kind of
effect, for example, the presence of a thin lubricant coating such as on a
storage media surface, catheter tubing or syringe needles. Additionally,
certain materials like PTFE are especially prone to this kind of cracking.
I have always assumed that this was at least in part due to the inherent
"grain" structure of any sputtered coating.

A layer of osmium metal deposited in an OPC osmium plasma coater, see URL
http://www.2spi.com/catalog/osmi-coat.html
which is not a sputtered coating, has no grain size (at least no one has
detected a grain size to our knowledge) and seems to be much more resistant
to the "crazy-paving" structure pattern effect on the above mentioned
samples. A comparison between different coating materials vs. osmium metal
is shown on URL
http://www.2spi.com/catalog/comparison-coating-results.html I propose
that the osmium coating approach would qualify for the requested "cure".

Disclaimer: SPI Supplies is the worldwide distributor of the OPC line of
osmium plasma coaters outside of Japan so we would naturally have a vested
interest in promoting the use of the osmium coaters for SEM sample
preparation.

Chuck

============================================

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

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






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From: gary-at-gaugler.com
Date: Mon, 28 Nov 2005 23:35:03 -0600
Subject: [Microscopy] Re: Thanks (TEM) + Question (SEM)

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

I've seen this "spider man" appearance before and attribute
it to sputtering at too high of pressure and too high of
current. Au is not a great metal for high resolution SEM
anyway...IMO. Au/Pd is better, and then consider Pt or Ir.

But it seems to me that low vacuum (15mT) and low current
(9mA) makes a big difference. But you will not likely get
15mT or lower without a turbo pumped system.

Try your system at the lowest vacuum you can get and see
what happens.

gary g.


At 12:20 PM 11/28/2005, you wrote:



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From: mike.reedy-at-cellbio.duke.edu
Date: Tue, 29 Nov 2005 16:14:07 -0600
Subject: [Microscopy] RE: Re: SEM imaging of bacteria encapsulation

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

Hi,
The 1991 BJ item is a meeting abstract, not article. It did cite the
prior work that inspired extensive trials we made of these methods in
the late 1980s.

Today I scanned it and a companion abstract about TAOS and TAURAC
fixation into a PDF and will separately email to you directly (no
idea how or if to transmit attachments via ListServer). My Acrobat 6
OCR function refuses to produce searchable text on these so they are
just images.; if i get an OCR to work, I will send that later.

David Popp and I planned to write up a methods paper with some
results but never finished the effort. We found that TAURAC-fixed
Araldite-embedded fibers of permeabilized, insect flight muscle
(fixed by TAURAC with a glutaraldehyde fixation interposed between
the TA and the UrAc fixes) still give fiber x-ray patterns showing
axial reflections out to 1.3 nm, and this result we will report in a
paper soon to be submitted by Kasim Sader (Univ. Leeds) et al.

The method can be cited from a 1994 peer-reviewed paper by H. Schmitz
et al., where it was described and some results illustrated in
Biophysical Journal (67:1620-1633).

-mike reedy-


At 10:26 AM -0600 11/29/05, Tindall, Randy D. wrote:
} Hi,
}
} I read this great post with great interest and have succeeded in
} tracking down two of the refs. I have been unable to locate the 1991
} Biophysical Journal article, however. Is the reference correct? Do you
} maybe have a PDF of this you could share? We have a client who is
} always looking for nifty ways to preserve biofilms and such.
}
} Thanks and Happy Holidays,
} Randy
}
} Randy Tindall
} EM Specialist
} Electron Microscopy Core Facility---We Do Small Well!
} W122 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
}
}
}
} -----Original Message-----
} From: mike.reedy-at-cellbio.duke.edu [mailto:mike.reedy-at-cellbio.duke.edu]
} Sent: Thursday, November 17, 2005 1:30 AM
} To: Tindall, Randy D.
} Subject: [Microscopy] Re: SEM imaging of bacteria encapsulation
}
}
}
}
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--
-mike reedy-

************************
Michael K. Reedy, M.D.
Duke Univ. Med. Center
Dept. Cell Biology, Box 3011 (for U.S. Mail)
458 Alex Sands Bldg, Research Dr (courier)
Durham, NC 27710

Office 919-668-2534
Lab 919-684-5674
Fax 919-681-9929
mike.reedy-at-cellbio.duke.edu
http://note.cellbio.duke.edu/Faculty/Research/Reedy.html

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From: jafarhan-at-rci.rutgers.edu
Date: Tue, 29 Nov 2005 20:32:45 -0600
Subject: [Microscopy] Atomic positions for WO2.9

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


Dear All,

I am doing some research with Tungsten Oxide. I got WO2.9 phase which is
tetragonal phase with space group of P4/nmm (from XRD). I wonder if
anybody have the atomic positions and Wykoff notations for this phase. I
have searched the literature but no luck.
Thank You

Jafar


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From: walck-at-southbaytech.com
Date: Wed, 30 Nov 2005 10:54:50 -0600
Subject: [Microscopy] Atomic positions for WO2.9

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

I know that there are a number of WO3 and WO(3-x) compounds in the PDF
database from when I worked on the oxidation of WS2 thin films. Have
you checked whether this space group agrees with the WO3 phase? If it
does, then the positions are the same and you have to think of the x=.1
as oxygen vacancies that are distributed through the material.

-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
walck-at-southbaytech.com


-----Original Message-----
X-from: jafarhan-at-rci.rutgers.edu [mailto:jafarhan-at-rci.rutgers.edu]
Sent: Tuesday, November 29, 2005 6:37 PM
To: Walck-at-SouthBayTech.com


Dear All,

I am doing some research with Tungsten Oxide. I got WO2.9 phase which
is tetragonal phase with space group of P4/nmm (from XRD). I wonder if
anybody have the atomic positions and Wykoff notations for this phase.
I have searched the literature but no luck. Thank You

Jafar


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From: mczech-at-sjm.com
Date: Wed, 30 Nov 2005 15:33:01 -0600
Subject: [Microscopy] viaWWW: Eye Damage by Light intensity?

Contents Retrieved from Microscopy Listserver Archives
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Email: mczech-at-sjm.com
Name: Mike Czech

Organization: St. Jude Medical, Inc

Title-Subject: [Filtered] Eye Damage by Light intensity

Question: Are there any studies that relate light intensity to eye damage from long term microscope usage?

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

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From: jmkrupp-at-cats.ucsc.edu
Date: Wed, 30 Nov 2005 18:36:10 -0600
Subject: [Microscopy] comparing images

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

I have an unusual question, I will try to explain it as best I can.

A researcher has asked me to find out if it is reasonable to do anything
like a quantitative comparison between fluorescence images of different
samples.

She has thick sections, 20 um, on a confocal scope. She sees differences by
eye between control and treatments, and she would like to quantify these
differences in some way.

We have thought about setting up the confocal to record the brightest
image, then without changing the settings, record an image of the other
slides. The idea would be that she could do something like compare the
brightness levels between them.

I don't know enough about other options or if this will even work to help her.

Does anyone do anything like this or is this not realistic.

Thanks

Jon

Jonathan Krupp
Microscopy & Imaging Lab
C230 Earth & Marine Science
University of California
Santa Cruz, CA 95064
(831) 459-2477
jmkrupp-at-cats.ucsc.edu



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From: donc-at-asmicro.com
Date: Wed, 30 Nov 2005 22:27:32 -0600
Subject: [Microscopy] SEM calibration standard

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

I'm soliciting the opinion of practicing SEM users regarding the materials
to use for a proposed high-precision 200 to 300 nm two-dimensional pitch
standard. The standard would be used to calibrate the horizontal scale (X
and Y axes) of a SEM or AFM. As I am an AFM expert, I already know what I
want in terms of the 3Dimension topography: the specimen will consist of an
array of bumps, with height probably 30-70 nm.
We are considering using etched silicon, silicon oxide on silicon, or
silicon nitride. Do any of these materials have advantages or disadvantages
for SEM use? Are some materials ok for low voltage ( { 1kV) and not ok for
high voltage?
Would it be a good idea to apply a metal coating? It seems obvious to me
that all magnetic materials should be avoided, such as Ni, Fe, Cr, Co. Are
there additional materials that are forbidden in some environments? For
example, I have heard that certain other metals are not allowed in some IC
fabs, but I don't know what they are.

I will be grateful to receive your input either via the list (which should
stimulate some good discussion) or offline.

regards,
Don Chernoff
==================================
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.]



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From: gary-at-gaugler.com
Date: Wed, 30 Nov 2005 22:54:09 -0600
Subject: [Microscopy] Re: SEM calibration standard

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

Look at the Geller 3 and 4 standards. They ought to
do what you need. If not, ask them for something else.

gary g.



At 08:47 PM 11/30/2005, you wrote:



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