stereology:preventing tissue shrinkage

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From:Donna Simmons <>
Date:Wed, 22 Sep 1999 16:03:06 -0700
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Dear HistoNetters,
I'm posting this question for a colleague, so could you email direct
replies to her?
I've told her a bit about HistoNet, and asked her to keep a synopsis of
private replies to share with the group when she solves her problem.
thanks, -Donna
This is a question regarding treatment of 40um frozen
sections of rat brain: immuno-stained while free floating,
and then mounted on slides after staining and air dried
before coverslipping and analysis...

>I have a question about immunohistochemistry. We have
>obtained equipment and software to do stereology but
>found that the immuno tissue I prepare has dehydrated
>from the original 40um sections to about 5 or 6 um. We
>cannot do stereology on these thin sections. Do you
>know any techniques to use during immuno, or for
>mounting the tissue after staining, that will have a
>less drastic effect on the volume of the tissue? I've
>heard only one suggestion so far. That was to mount
>the tissue on a slide then coverslip without
>dehydrating in either alcohol or zylene, then seal the
>coverslip edges with something like fingernail polish.
>Do you have any experience with this kind of problem,
>or do you know any labs doing stereology on
>immuno-treated tissue?
>M. Claire Cartford, Ph.D.  <>
>University of Colorado Health Sciences Center
>4200 E. 9th Ave.
>CPH Box C268-71
>Denver, Colorado 80262
>tel. (303)399-8020 ex 2384
>fax  (303)331-8324

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