[Histonet] IF Protocol on skin

Gagnon, Eric gagnone <@t> KGH.KARI.NET
Fri Feb 13 09:34:47 CST 2009

Hi Angela,
We used Ventana FITC Primary Antibodies.


From: Angela Bitting [mailto:akbitting <@t> geisinger.edu]
Sent: Wed 2/11/2009 5:11 PM
To: Gagnon, Eric
Subject: Re: [Histonet] IF Protocol on skin

Hi eric,
You said you experimented with using the FITCs on your XT. Was that using Dako antibodies or Ventana's?

Angela Bitting, HT(ASCP)
Technical Specialist, Histology
Geisinger Medical Center
100 N Academy Ave. MC 23-00
Danville, PA 17822
phone  570-214-9634
fax  570-271-5916
>>> "Gagnon, Eric" <gagnone <@t> KGH.KARI.NET> 02/11/09 4:02 PM >>>
Hi Gudrun,

Our IF protocol is very similar to yours...we use 1/30 dilutions as well, although we rinse before the FITC-antisera (Dako) are applied, with frozen sections cut at 5 microns. 

We did experiment with this procedure on our BenchMark XT's, with some success.  The pathologist who reads most of renal biopsies IF's decided that our manual method was "robust" enough (i.e. foolproof enough) that we could continue using it.  Manual staining saves us trying to fit in a new or current run when the XT is already running.

Regarding our dilutions, we make 3 ml at a time, and have not had any problems with the staining diminishing before the antisera are used up.

Eric Gagnon MLT
Histology Laboratory
Kingston General Hospital
Kingston, Ontario, Canada

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