[Histonet] Dilemma.....

From:brucea@unimelb.edu.au, hickford@unimelb.edu.au

Hi everyone
I posted a question on the histonet a week ago but I don't think I=20
explained my dilemma clearly enough. I am using alkaline phosphatase=20
histochemistry (not immunohistochemistry!) on PFA-fixed whole 
marsupial blastocysts and embryos to detect primordial germ cells=20
(PGCs). I use 1mg/ml Fast Blue salt and 1mg/ml Naphthol AS phosphate=20
sodium salt in 0.2M Tris buffer, pH 9.4 for the histochemistry, a=20
standard method for the detection of PGCs. The staining is good but I=20
have been told that it is not permanent. Ideally, after staining 
these embryos histochemically for PGCs, I would like to dehydrate,=20
paraffin- embed, section and then stain them immunohistochemically=20
(ie, with antibodies) for other germ cell markers and several growth=20
factors. Does anyone know of a histochemical method for the detection=20
of germ cells that would later permit me to do this? We have 
well-established methods for immunohistochemistry for several germ=20
cell markers in our lab. Thank you to the people who took the time to=20
help last time,
Dr Danielle Hickford
Research Fellow
Department of Zoology, University of Melbourne,
Australia 3010.

BRUCE ABALOZ                           PH:61383446282
HISTOLOGIST                            FAX:61383447909
DEPT.of ZOOLOGY                        EMAIL: brucea@unimelb.edu.au

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