[Histonet] gelatin embedding

MKing making <@t> ufl.edu
Mon Aug 6 08:44:08 CDT 2007

This works pretty well, just be careful about potential interactions 
with immuno epitopes, especially near the tissue surface.  Contact me if 
you have questions,
Mike King
UF Pharmacology & Therapeutics

Gelatin-Albumin Embedding
modified from  Levin M. A novel immunohistochemical method for 
evaluation of antibody specificity and detection of labile targets in 
biological tissue. J Biochem Biophys Methods. 2004 Jan 30;58(1):85-96.

22.5  ml PBS
1.1 g gelatin
225 ml dH2O
heat to 60 deg. C., dissolve completely
cool to room temp., add 67.5 g egg albumin (bovine albumin ok), 
dissolve, aliquot, store frozen

to use:
thaw, add 420 ul formalin (37% formaldehyde) to 1.5 ml gelatin-albumin, 
mix thoroughly.
pour into the bottom of molds over ice and allow to set (> 1 hr.).
mix 2nd batch of formalin (1.5-3 ml for mouse brain) & gelatin-albumin, 
take specimen from 30% sucrose PBS, blot with Kimwipe, gently stir in 
formalin/gelatin-albumin, pour into mold and orient.  allow to set, then 
spatula block out of mold, trim, equilibrate in 30% sucrose PBS, and 
section frozen.

210 ul glutaraldehyde can be used instead of formalin, but will react 
faster and may impair immunoreactivity more.

Date: Fri, 3 Aug 2007 13:09:23 -0400
From: "Joshua Berman" <jb481 <@t> columbia.edu>
Subject: [Histonet] A flurry of questions about gelatin embedding,
	freezing,	and mounting for mouse brain floating sections.
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <001a01c7d5f1$0a863270$3926a8c0 <@t> JoshB>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";

I am soliciting opinions/advice about floating section ICC in mouse 
brain. i know this is probably pretty basic stuff, but any help will be 
greatly appreciated...

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