RE: formalin substitutes

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From:"Greer, Patricia" <>
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We have used the following fixative for demonstrating CD3, CD4, and CD8.  We
compared the staining for these cell markers after fixing tissue in formalin
and in the JB fixative.  We could not demonstrate the cell markers after
formalin fixation but they stained really well after fixation in the JB
fixative.  Be careful not to overfix in this - the tissue will become very
hard and brittle if it stays in the JB fix too long.  After a maximun of 24
hours, we transfer the tissue to 70% alcohol.

		JB Fixative

	0.1 M  Tris Buffer, pH 7.4		1000ml
	Calcium acetate				0.5 g
	Zinc Acetate				5.0 g
	Zinc Chloride				5.0 g

Mix to dissolve.  The final pH will be approximately 6.8 (6.5-7.0).  Do not
readjust the pH, as this will cause the zinc to come out of solution.  Small
amounts of insoluable zinc oxychloride often contaminate zinc chloride.
This may result in a very small amount of white precipitate.  This will not
cause any problems but can be removed by filtering the solution.
Fix tissue for 2-24 hours.

Beckstead JH. A Simple Technique for Preservation of Fixative-sensitive
Antigens in Paraffin-embedded Tissues: Addendum (Letter to the Editor). J
Histochem Cytochem 1994; 43:345

Pat Greer
Centers for Disease Control
Infectious Disease Pathology   

-----Original Message-----
From: []
Sent: Wednesday, June 07, 2000 2:43 PM
Subject: formalin substitutes

What non-formaldeyde based fixers are people using to preserve antigens in
paraffin sections? I have been using zinc formalin with little improvement
and am looking for something else. Does anyone used Shandon's Glyo-Fixx?
Are there tried and true ones out there?

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