Re: [Histonet] Eosinophil marker

From:John Kiernan

You identify yourself only as "Unipath IHC."
Why? Are you a commercial outfit asking for
free advice on the internet? If so, would
you sell services to clients on the strength
of someone's 15-minute response on a
listserver? Internet replies, including mine
(which follows) have no real value.  If you
do work for a client based on advice collected
from an internet enquiry you are asking to be

That said, I'll offer some advice. do not 
follow it without checking that it's based
on established and generally accepted 
knowledge. Compare 3 major textbooks. If
there are discrepancies, follow up the
references in your library. Get photocopies
of older papers by interlibrary loan if
necessary. That's one of the ways to get
details of techniques.

Antibodies are expensive. Eosin isn't, and it
stains the cytoplasmic granules of eosinophils 
strongly. If you stain with an alkaline (pH 9)
aqueous solution of eosin, the only red things
in animal tissues will be eosinophil granules,
Paneth cell granules (in the intestine) and
tails of spermatozoa (if present). I think this
is a complete list; someone, please correct me 
if I've missed something.

If you need a counterstain, use any
haemalum (Ehrlich, Gill, Harris, Mayer etc)
or a blue basic dye (eg toluidine blue at 
pH 4) according to your needs. Either of
these "counterstains" should probably be 
done before the alkaline eosin staining.

Dyes are cheaper and easier to use than
antibodies!  Immunohistochemistry is an
important family of methods, but it is 
expensive and is not needed for simple jobs
such as staining eosinophils. Your 
enquiry did include information about
the tissue you work with. Whatever it
is, you should not confuse eosinophils
with Paneth cells or sperm tails!
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
UniPath IHC wrote:
> Can anyone recommend an eosinophil antibody that works in FFPE tissue?
> _______________________________________________

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