[Histonet] detect primordial germ cells in PFA-fixed marsupial embryos...?

[Bruce Abaloz]

Hi
I have been using alkaline phosphatase histochemistry to detect 
primordial germ cells in PFA-fixed marsupial embryos. I use 1mg/ml 
Fast Blue salt and 1mg/ml Naphthol AS phosphate sodium salt in 0.2M 
Tris buffer, pH 9.4. The staining is good but I have been told that 
it is not permanent. Ideally, I would like to dehydrate these stained 
specimens, embed them in paraffin and then section and stain them 
immunohistochemically for other germ cell markers and several growth 
factors. Does anyone have a protocol utilising alkaline phosphatase 
histochemistry that would allow me to do this? Thanks for any advice 
in advance,

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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