This works pretty well, just be careful about potential interactions
with immuno epitopes, especially near the tissue surface. Contact me if
you have questions,
UF Pharmacology & Therapeutics
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
thaw, add 420 ul formalin (37% formaldehyde) to 1.5 ml gelatin-albumin,
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
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"
Subject: [Histonet] A flurry of questions about gelatin embedding,
freezing, and mounting for mouse brain floating sections.
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
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