[Histonet] Dilemma.....

[brucea <@t> unimelb.edu.au]

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 <@t> unimelb.edu.au


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

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