RE: [Histonet] Dilemma.....
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
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
School of Graduate Medical Sciences
Podiatric Medicine and Surgery
Miami Shores, Florida
[mailto:firstname.lastname@example.org] On Behalf Of
Sent: Monday, May 10, 2004 9:59 PM
Subject: [Histonet] Dilemma.....
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
Department of Zoology, University of Melbourne,
BRUCE ABALOZ PH:61383446282
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