Hi Josie,

We routinely fix cytospins in cold acetone for 10 minutes but you may want
to do some comparison studies using methanol and 10%NBF as well.  We have
found that letting the cytospins sit in APK for awhile will lyse the red
blood cells and really help in cutting down on endogenous peroxidase
staining.  Also, instead of using a "Frozen", "Wetload" protocol, using a
"Paraffin" protocol will include application of the inhibitor which will
further cut down on endogenous peroxidase staining.  We also always use AB
block on frozens and cytospins.

Linda A. Sebree, HT
University of Wisconsin Hospital & Clinics
Immunohistochemistry/In Situ Hybridization Laboratory
D4/218-2472
600 Highland Avenue
Madison, WI  53792-2472


(608)265-6596
FAX: (608)263-1568

> -----Original Message-----
> From:	Josefa Nava [SMTP:pepanava@hotmail.com]
> Sent:	Thursday, September 07, 2000 4:50 PM
> To:	histonet@pathology.swmed.edu
> Subject:	Re: Daily Digest
>
> Can someone tell me the best way to prepare cytospin , smears for
> immunostain using Ventana Nexes . I am interested on what kind of fixative
>
> to use and how to prevent red blood cell staining or endogenous background
> .
> I am using Avidin /Biotin blocker but it is not enough to remove all the
> background staining. Any suggestion is very much appreciated.
>
> Thank you.
> Josie
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