RE: use of zinc formalin for antigen preservation?

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From:"Greer, Patricia" <pwg1@cdc.gov> (by way of histonet)
To:histonet@histosearch.com
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We have used a zinc fixative (JB fixative) for mouse tissues prior to
immunostaining for CD3, CD4 and CD8. It does allow demonstration of these
cell markers which we could not be demonstrate in formalin fixed tissues.
We did not put it on the tissue processor; we fix all our tissues prior to
processing and start our processing at 70% alcohol. I wouldn't think that
this particular fixative would be a good choice for routine fixation.

The formula for the JB fixative we used is:
    0.1M Tris buffer, pH7.4     1000ml
    Calcium acetate              0.5g
    Zinc Acetate                 5.0g
    Zinc Chloride                5.0g
Mix to dissolve.  The final pH will be approximately 6.8.  Do not readjust
the pH, as this will cause the zinc to come out of solution.
Fixation should be no longer than 48 hours, otherwise the tissue is brittle
and impossible to cut.

Reference:
Beckstead JH: A simple technique for preservation of fixation-sensitive
antigens in paraffin-embedded tissues. The Journal of Histochemistry and
Cytochemistry 1994;42:1127
Beckstead JH: Letter to the editor. Journal of Histochemistry and
Cytochemistry 1995;43:345 (This is the actual formulation of the fixative).

***********************
Patricia W. Greer, MT,HTL(ASCP)
Centers for Disease Control
Infectious Disease Pathology
Mail Stop G-32
Atlanta, GA 30333
phone: 404-639-2811



-----Original Message-----
From: Susie Seward [mailto:sseward@statlab.com]
Sent: Friday, November 19, 1999 3:44 PM
To: Histonet
Subject: use of zinc formalin for antigen preservation?


I have a physician inquiring about the use of zinc formalin for antigen
preservation,  and inquiring about using it on their tissue processor. He
said he wanted to avoid antigen degradation. Any zinc guru's please respond
with how and why you are using zinc formalin verse 10% NB formalin (or do
you use both?) I would appreciate it. Thanks.




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