Re: zinc fixative (non formalin)

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From:"Hiroaki Nitta" <>
To:"Gayle Callis" <>, <>
Date:Mon, 16 Aug 1999 23:08:42 -0400
Content-Type:text/plain; charset="iso-8859-1"

Dear All:
I have used this zinc solution that Gayle prefers to call "Zinc-Tris
fixative" when I was with GeneMedicine, Inc., The Woodlands, TX, now called
Valentis, Inc.  The reason why I used this solution was that I was informed
the Cancer Gene Therapy Program at GeneMedicine would be collecting so many
tumor samples (about 50 samples or more) from an experiment.  As most of you
know that scientists, like you are, would use acetone-fixed frozen sections
for in situ leukocyte detection.  Could you imagine to process more than 50
samples for preparing frozen sections at the same time?  We tested a lot of
antibodies on Zinc-Tris-fixed spleen sections and we found that the majority
of antibodies worked like on frozen sections.  This solution worked very
well on tumor samples, too.  As Gayle mentioned, this solution did not work
on bone samples when I worked for another biotechnology company called
Osiris Therapeutics, Inc., Baltimore, TX.  I don't recommend for muscle
samples, either.

I would love to hear your experience with this fixative for
immunohistochemistry.  Please let me know.

Hiro Nitta

PS  I am going to accept a job pretty soon after being laid off by Osiris.
I will let you know who I am going to work for when I decide.  I have 4 job
offers on table now:-)

-----Original Message-----
From: Gayle Callis <>
To: <>
Date: Friday, August 13, 1999 3:04 AM
Subject: zinc fixative (non formalin)

>A word of caution about zinc fixative (not to be confused with zinc
>Here are original references on this fixative, developed by J Beckstead.
>Nitta et al used alk phos substrate.  Endogenous peroxidase cannot be
>quenched, I couldn't do it, others have reported the same.  Some people
>have had problems with fixation times, pays to do a fixation time study,
>some did 8 hours or less, and others went longer, recommended is 24 hours,
>up to 3 days.  Because this was in a buffer, I stored mine in a
>and preferred to call it ZincTRIS fixative, to not confuse it it with zinc
>formalin.  A nice fixative, but you cannot decalcify bone, swells terribly,
>It tends to cause more friable sections, just soak them a bit after
>on ice water.
>H. Nitta et al Cell Vision 1997 4:73-80  Improved in situ
>immunodetection of leukocytes on paraffin embedded mouse spleen
>J Histochem Cytochem 42:1127-1134, 1994  J.H.Beckstead A simple technique
>preservation of fixation sensitive antigens in paraffin embedded
>J Histochem Cytochem 43:345 JH Beckstead  Same title (addendum)
>Gayle Callis

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