RE: IHC followed by PAS counterstain

From:Tony Henwood

RE: IHC followed by PAS counterstain


On paraffin sections we stained for the antigen first (using peroxidase-DAB) followed by Periodic acid-Thionin Schiffs. This gave Blue PAS positive material with Brown antigen localisation - very pretty.

Fixation will depend on the antigen being localised. Try immediate fixation in 95% ethanol for 5 minutes and air-drying followed by cold acetone. Some antigens prefer one fixation over another (as if they have any choice!!!)

Hope this helps,

Tony Henwood JP, BappSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital at  Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: (02) 9845 3306
Fax: (02) 9845 3318

-----Original Message-----
From: Robin Thain []
Sent: Saturday, 21 September 2002 7:32
To: Histonet
Subject: IHC followed by PAS counterstain

Has anyone out there done immuno and then counterstained with a PAS instead
of hematoxylin.  We want to do this on frozen sections.  I need info about
fixatives, method.  Those kinds of things.  TIA.

Robin Stocks
University of Alberta

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