[Histonet] IHC controls for Cytology

Sebree Linda A. la.sebree <@t> hosp.wisc.edu
Wed Dec 14 09:08:23 CST 2005

Hi Becky,

When we were doing IHC on cytospins and ThinPreps, we used frozen tissue
sections that had been acetone fixed the same way as the cytology preps.
We were able to optimize our antibodies using the frozen sections and
the protocols developed always worked for the cytology preps as well.
We figured frozens were a much closer preparation methodology than
paraffin sections and our pathologists were happy with the compromise we
came up with.

Linda A. Sebree
University of Wisconsin Hospital & Clinics
IHC/ISH Clinical & Research Laboratory
600 Highland Ave.
Madison, WI 53792
FAX:  (608)262-7174

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Orr,
Sent: Wednesday, December 14, 2005 8:44 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] IHC controls for Cytology


Hi everyone,

We have very recently begun to destain pap stained smears as  well as
unstained  cytology smears,  and run IHC.

Because my Docs are happy with the results, I anticipate receiving more
of these types of smears to run.

I only  have FFPE tissue in stock to use as control, so I give them the
slides with the disclaimer that the control has been treated differently
than the smear.

This seems to be acceptable as long as they can detect some positive
stained cells (TTF, PanCk, WT-1  Mart-1 for example). But if the smear
is negative, how can I be sure it's a true negative?  My antibodies
aren't really titered for  smears....

I would appreciate some comments from anyone who is running a
standardized protocol for smears. Do you have a set of smears you can
use as controls?  I have access to our molecular lab and there is
potential to make my own cell cultured controls,  what do you think?


Many thanks,



Becky Orr CLA,HT(ASCP)

IHC Lead 

Evanston Northwestern Healthcare



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