The old Gomori method for alkaline phosphatase left a an indestructible
black deposit of cobalt sulfide.  Gomori's method always gave me good
results.  The method is given in Lillie's HISTOPATHOLOGIC TECHNIC AND
PRACTICAL HISTOCHEMISTRY.  The 4th edition is out of print and unavailable,
but a used copy of the 2nd or 3rd edition can be had from Barnes and Noble
or Amazon at a reasonable price.
   The current (3rd) edition of Kiernan's HISTOLOGICAL AND HISTOCHEMICAL
METHODS gives a cerium-DAB-nickel method that leaves a quite durable
precipitate.
   Either method allows clearing in xylene and mounting in resin.  I don't
know how either would affect subsequent immunohistochemical reactions.

Allen A. Smith, Ph.D.
Professor of Anatomy
Barry University
School of Graduate Medical Sciences
            Podiatric Medicine and Surgery
Miami Shores, Florida


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
brucea@unimelb.edu.au
Sent: Monday, May 10, 2004 9:59 PM
To: histonet-bounces@lists.utsouthwestern.edu;
histonet@lists.utsouthwestern.edu
Subject: [Histonet] Dilemma.....


Hi everyone
I posted a question on the histonet a week ago but I don't think I 
explained my dilemma clearly enough. I am using alkaline phosphatase 
histochemistry (not immunohistochemistry!) on PFA-fixed whole 
marsupial blastocysts and embryos to detect primordial germ cells 
(PGCs). I use 1mg/ml Fast Blue salt and 1mg/ml Naphthol AS phosphate 
sodium salt in 0.2M Tris buffer, pH 9.4 for the histochemistry, a 
standard method for the detection of PGCs. The staining is good but I 
have been told that it is not permanent. Ideally, after staining 
these embryos histochemically for PGCs, I would like to dehydrate, 
paraffin- embed, section and then stain them immunohistochemically 
(ie, with antibodies) for other germ cell markers and several growth 
factors. Does anyone know of a histochemical method for the detection 
of germ cells that would later permit me to do this? We have 
well-established methods for immunohistochemistry for several germ 
cell markers in our lab. Thank you to the people who took the time to 
help last time,
Dr Danielle Hickford
Research Fellow
Department of Zoology, University of Melbourne,
Australia 3010.
hickford@unimelb.edu.au


-- 
BRUCE ABALOZ                           PH:61383446282
HISTOLOGIST                            FAX:61383447909
DEPT.of ZOOLOGY                        EMAIL: brucea@unimelb.edu.au
THE UNIVERSITY Of MELBOURNE.
VICTORIA.AUSTRALIA 3010

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permission.  -  Eleanor Roosevelt
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