Hi Bob,
I do 30-50 micron cryosections of skin and mount them on charged slides. We
have had success with both flourescence and standard immunostaining (have not
done PGP), but we incubate the primary overnight for everything!!
Skin has a nasty habit of separating at the dermal/epidermal junction and I 
think you are asking for grief moving free floating sections around (been
there, done that).
Good luck, Marge Lehman

-----Original Message-----
From:	Neuropathology [SMTP:Neuropath.Frenchay@dial.pipex.com]
Sent:	Thursday, March 22, 2001 9:25 AM
To:	Histonet
Subject:	Immuno on freezing microtome sections.

I 've been asked to cut 50 micron freezing microtome sections of a skin
biopsy and immunostain with PGP 9.5. I'm happy (??!!) to cut the sections
but would welcome any advice on the immunostaining. Is it better to free
float the sections through the reagents or mount the sections onto slides,
air dry and then stain? Is immunofluorescense best or can I use the standard
immuno method as used for cryostat sections?


Bob Quilty
Dept of Neuropathology
Frenchay Hospital
Bristol
UK