[Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee & Peggy/Tony, SOMEONE....)

[Bruce Abaloz]

Hi, my name is Danielle & I need some advice PLEASE -
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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-- 
BRUCE ABALOZ                           PH:61383446282
HISTOLOGIST                            FAX:61383447909 
DEPT.of ZOOLOGY                        EMAIL: brucea <@t> unimelb.edu.au
THE UNIVERSITY Of MELBOURNE.
VICTORIA.AUSTRALIA 3010           

               Nobody can make you feel inferior without your permission.  -  Eleanor Roosevelt
                         DANCE LIKE NO-ONE'S WATCHING          ********************************************************************************                                                                  This electronic message and all contents contain information which may be
privileged, confidential or otherwise protected from disclosure.The information is intended to be for the addressee only. If you are not the addressee, any disclosure, copy, distribution or use of the contents of this message is prohibited. If you have received this electronic message in error, please notify us immediately and destroy the original message and all copies.