RE: [Histonet] Zinc Formalin and Iron stains

From:"Liz Chlipala"

Gayle is correct if the zinc formalin is not buffered.  When I initially
worked with this fixative in the mid eighty's it was not commercially
available.  If you made this solution up from scratch its pH was between 1
and 2, so very acidic. I believe you can only purchase the buffered form
through a vendor and you can't make it up from scratch, the zinc will come
out of solution when you try to make it up in a buffer.  Iron can be removed
in acid solutions, that's why it can be sometimes difficult to see iron in
decalcified bone marrow cores.  Just search the archives for iron and bone
marrow and you'll see lots of responses.  Peggy Wenk has a nice post that
goes over what you need to do to retain the iron in bone marrow specimens,
she also states in that post that iron can be removed in fixatives that are
acidic and to make sure the fixative that you use is buffered and is at the
correct pH.


Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, CO 80308
phone (303) 735-5001
fax (303) 735-3540
Ship to Address:
Premier Laboratory, LLC
University of Colorado at Boulder
MCDB, Room A3B40
Boulder, CO 80309

-----Original Message-----
[] On Behalf Of Gayle Callis
Sent: Monday, April 16, 2007 12:59 PM
To: Tarango, Mark;
Subject: RE: [Histonet] Zinc Formalin and Iron stains

We have used zinc formalin, although it was Anatech's buffered zinc 
formalin formulation, and had wonderful results on human kidney biopsies - 
excellent morphology and no exploded cells that the pathologist could 
see.  He loved the results.   The sections cut at 1 - 2 um.  When we tried 
this fixative on animal tissues, we also had great success.  Could it be it 
was a zinc formalin that was NOT buffered correctly or even at all that 
caused the exploding cells?

As for iron staining after zinc formalin, not sure what the results are. 
However, you should speak to Anatech, in particular, Ada Feldman, she could 
let you know if iron staining is compromised by this fixative.

At 12:32 PM 4/16/2007, you wrote:
>I don't know, we only use 10% NBF for bone marrows here, but I've seen a
>video of some cells getting fixed with zinc formalin.  The cells were
>popping and exploding all over the place.  Made me think, "no wonder you
>get that great nuclear detail."

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610

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