[Histonet] RE: IHC free floating sections

[Mitchell Jean A]

Michael:  I use 1.5mm nichrome transfer loops for IHC staining of 50um
free floating sections and I move them into clean culture wells for each
solution.  I purchase mine from Ted Pella.  It does takes some practice
but the loops work quite well with minimal damage to the tissue.

Jean Mitchell, BS, HT (ASCP)
University of Wisconsin Hospital & Clinics
Neuromuscular Laboratory
600 Highland Avenue
Madison, WI  53792-5132 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Michael
Patrick
Sent: Friday, July 31, 2009 2:14 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] IHC free floating sections



I would like to stain free floating sections (fixed rat brain; 50um). Is
it necessary to transfer sections to "clean" culture wells with a paint
brush every time I wash or incubate with antibodies? I find this causes
significant damage to my tissue (especially since I do a lot of washes
etc.). Could I simply use a pipette to draw out the old solution and
then replace with the appropriate solution? What do most people do in
these situations?

Much thanks

Mike

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