[Histonet] Agar embedding protocol

Jackie M O'Connor Jackie.O'Connor <@t> abbott.com
Mon Dec 19 15:07:02 CST 2005

Have you tried Sally Schlessinger's "double embedding" method?  You embed 
multiple veins (or nerves or gut or anything long) longitudinally in a 
shallow amount of  paraffinin a  mold, then when the paraffin has 
hardened, you can cross section the longitudinal veins, and re-embed them 
at a 90 degree angle.  She wrote an article somewhere about this, about 10 
years ago, I think.  It works great. 

"Amy Porter" <portera <@t> msu.edu>
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
12/19/2005 02:51 PM
Please respond to Amy Porter

        To:     <histonet <@t> lists.utsouthwestern.edu>, <MVaughan4 <@t> ucok.edu>
        cc:     (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT)
        Subject:        Re: [Histonet] Agar embedding protocol

Mel - I know that this is an older posting, but I am just now getting 
through my bunches of histonet save items.  Are you using the agar for 
paraffin embedding or for frozens?  I missed the original query into what 
you are using this for.  Our laboratory is trying to embed one or multiple 

segments of 6-8 week old mouse mesenteric veins for cross sections!  We 
having a heck of a time with these and I am trying to find a method which 
will withstand automated processing and paraffing embedding.  If you or 
anyone else out there would have any suggestions that would be great. 
Thanks in advance for your responses and suggestions.
----- Original Message ----- 
From: <MVaughan4 <@t> ucok.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Friday, December 16, 2005 1:26 PM
Subject: [Histonet] Agar embedding protocol

> Jim,
> Bio-Rad agar 3% in water, boil to melt, let cool to about 60 degrees,
> quickly pour into a cryoembedding mold and orient sample before the agar
> gels.
> Once the agar hardens you can cut out the sample surrounded by agar.
> Mounting works best using a dissecting scope if you have a small sample.
> You might be able to use low-melt agar if your sample is sensitive to
> heat. The only problem I have noticed is that sometimes there is
> nonspecific background staining of the tissue or loss of antigen, 
> if the agar is too hot. You might test on an unimportant sample first.
> Mel
> Melville B. Vaughan, Ph. D.
> Assistant Professor
> Department of Biology
> University of Central Oklahoma
> Edmond, OK 73034
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