[Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee &
Peggy/Tony, SOMEONE....)
LORALEE GEHAN
lagehan <@t> hotmail.com
Tue Apr 27 15:05:15 CDT 2004
With my experience it won't survive the processing. Try it plastic.
The processing is a bit longer but the stain works nicely.
Hope that helps.
Loralee Gehan, HTL
>From: Margaret Blount <mab70 <@t> medschl.cam.ac.uk>
>To: 'Bruce Abaloz' <brucea <@t> unimelb.edu.au>,
histonet-bounces <@t> lists.utsouthwestern.edu,
histonet <@t> lists.utsouthwestern.edu
>Subject: RE: [Histonet] Re: HELP
PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee & Peggy/Tony, SOMEONE....)
>Date: Tue, 27 Apr 2004 10:46:25 +0100
>
>Hi, Everyone,
>
>With regard to this problem, I was wondering if the alk phos enzyme
would
>survive processing, I rather think it would otherwise we wouldn't
need to
>add levamisole when immunostaining paraffin sections using alk phos
as a
>label. So why not process into wax then do the detection of alkaline
>phosphatase and the other markers you wish to identify.
>
>What do other histonetters think? Is it worth a try?
>
>Margaret
>
>Margaret Blount
>Chief Technician
>Clinical Biochemistry
>University of Cambridge
>Addenbrooke's Hospital
>Hills Road
>Cambridge
>CB2 2QR
>
>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu
>[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Bruce
>Abaloz
>Sent: Tuesday, April 27, 2004 2:30 AM
>To: histonet-bounces <@t> lists.utsouthwestern.edu;
>histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] Re: HELP
PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee
>& Peggy/Tony, SOMEONE....)
>
>
>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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permission.
>- Eleanor Roosevelt
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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
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> DANCE LIKE NO-ONE'S WATCHING
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