[Histonet] Histo gel Issues

[McCormick, James]

All,
 with Histogel "drying","hardening", and the loss of specimens. 
I am particularly interested in artifacts of processing with algenates, among which is Histogel.  As I am analyizing the problem for our own use I have determined that there are variables introduced with the following:
1. Age of the product.  Has it been opened and lost moisture thru evaporation.
2. Type of solutions in the beginning of processing. Particularly a rapid ascent into higher alcohols, (they cause the gel to shrink and harden).
3. Heat exposure time in melting...was the heat controlled? Time?
I would be pleased to make specific comments if you will email the time and solutions in the dehydration steps,clearing solution and paraffin exposure times.
Interested, 
J.B.McCormick, M.D.
CSO Leica-Biosystems,
St. Louis, Mo 

 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jo Dee Fish
Sent: Tuesday, February 03, 2009 10:50 AM
To: 'pam plumlee'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Histo gel Issues

Dear Pam,
I've had the same problem.  I called Richard-Allen just after they were
"absorbed" by Thermo Fisher and got no answers.  They had never heard of
such a problem and didn't know how to solve it.  I have heard from another
user that had the same exact problem.  I lost precious samples, E6.5 mouse
embryos, because of this "drying out and hardening" of the histogel.  I had
to stop using it all together.
If anyone has any suggestions, please let us know!
Take care Pam and all histonetters,
Jo Dee 


~~Jo Dee Fish~~
Senior Research Technologist
The J. David Gladstone Institutes
Co-manager Histology and Microscopy Core
 
Telephone: (415) 734-2567
Fax: (415) 355-0824
E-mail: jfish <@t> gladstone.ucsf.edu
 
Mailing address:
The J. David Gladstone Institutes
1650 Owens Street
San Francisco, CA 94158

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of pam plumlee
Sent: Monday, February 02, 2009 1:06 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Histo gel Issues


Dear Group: I'm having problems with the processing of small tissues in
histogel. I follow the suggested embedding directions on the package and
then process the gel blocks.  The results are very inconsistent-ideally,
I'll get nice soft gel blocks-but, usually in a batch of 10 I get 2 good
blocks and 8 dried up, flat hard squares.  They are all handled and
processed the same.  Anyone experience this before?  Thanks for any input.

Pam Plumlee H.T.
Pfizer La Jolla
pam.plumlee <@t> pfizer.com


      

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