Re: Methacrylate

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From:Karen S Pawlowski <kna101@utdallas.edu> (by way of histonet)
To:histonet@histosearch.com
Reply-To:
Date:Mon, 31 Jan 2000 22:47:19 -0500
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Peter,

One thing you could try is using the methacrylate that you usually use for
hard tissues and etch the sections to expose the antigens prior to any
other manipulation of the section in your immunohistochemical protocol.
One solution to do this with is a mixture of sodium hydroxide in methanol
that has been allowed to "cure" for a few weeks in a brown bottle.
It's an old technique and maybe someone can give us the reference, as
I seem to have lost it.  The concentration of the stock solution should be
5-10 gm of NaOH to 50 ml methanol.  This stuff is very caustic so keep it
in a hood and handle with gloves.
There are probably people out there who have used this for plastic
sections that could give you more info and they probably have suggestions
on some less harsh ways to do it too.
I've been using it for a technique I'm developing to etch celloidin
sections at light level.  I make up a working solution of 1 to
100 - 1 to 500 in methanol. The longer the stock sits, the stronger it
gets.  The sections will have to be in this for anywhere from 10 minutes
to 30 minutes and I rinse in distilled water to stop the reaction.

Good luck,
Karen Pawlowski
Sr. Res. Tech./UT Southwestern Med. Ctr.
PhD Candidate/UT Dallas,
Dallas, TX


On Wed, 27 Oct 1999, Rippstein, Peter wrote:

> Would anyone have a methacrylate protocol which will accomodate the cutting
> of hard tissue and subsequent immunohistochemistry. We are working with
> stented arteries. Any suggestions would be greatly appreciated.
>
> Peter Rippstein
> Department of  Anatomical Pathology
> Ottawa Hospital (Civic Campus)
> Ottawa,Ontario,Canada
>
>




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