Re: fix with IHC on brain

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From:"John C. Dennis" <dennijc@vetmed.auburn.edu>
To:lynn_brennan@smtplink.Coh.ORG
Reply-To:
Date:Fri, 9 Jul 1999 09:20:20 -0500 (CDT)
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Lynn

I think there is no "best" fixative for IHC with any tissue.  I am
thinking these days that everything depends on the antibody you want to
use and its target.  I look at rodent brain and olfactory tissue.  We fix
with Bouin's, 4% buffered paraformaline, and various combinations of
glutaraldehyde and paraformalin.  Some antibodies work with some of the
fixatives that other antibodies do not work with at all (if that sentence
makes sense at all).  You'll need to find the combination that best
suits your needs and then away you go.

The good people on the histonet have told me lately that cold acetone does
real well for unfixed frozen tissue.  Brain is appearently obdurate by
tradition and doesn't stick on slides but we've remedied that by dunking
our Superfrost Plus slides in liquid poly-l-lysine (Sigma).  It works
well.  The coating idea also came from the good people plugged into the
histonet, by the way.

Hope this helps.

Cheers



John Carroll Dennis
Anatomy, Physiology, and Pharmacology
109 Greene Hall
Auburn University, AL  36849


On Wed, 7 Jul 1999 lynn_brennan@smtplink.Coh.ORG wrote:

> What is the best fixative (you can make in the lab) using IHC and frozen
> sections on embryonic spinal cords?
> Any suggestions for the best fixative with any brain tissues?
> Many thanks,
> Lynn Brennan
> Division of Neurosciences
> City of Hope  Medical center
> 1500 Duarte Road
> Duarte,California  91010
> 
> 




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