RE: [Histonet] Immuno's on Thin Prep specimens

From:"Sebree Linda A."



I disagree Rene in terms of using a paraffin section as a positive control.  Your two specimens (thin prep & + control) are not prepared the same.  In order to get your + control to stain , you may have to employ HIER which as you state you would not do with the thin prep.  That pretreatment of the + control makes it even "more different" than your patient unknown.  I recently stuck my neck out at work for this very same issue.  I refused to be part of running a cytospin with a FFPE staining protocol and + control.  Our lab used to run cytospin IHC protocols and many times had to adjust the FFPE protocols to work for the cytopsins.  At that time we used frozen section tissue controls for our positives for lack of a better alternative.  These came closer to the patient specimen than FFPE controls and actually stained identically to the cytospins.

Just my experience.

Linda Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-3224
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX: (608)262-7174


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Friday, September 28, 2007 7:49 AM
To: AWeiss@shorememorial.org; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Immuno's on Thin Prep specimens


If using thin preps for IHC, you do NOT need to do HIER because they are acetone/air dried fixed, not formalin fixed. The rest of the IHC protocol is the same as for a FFPE tissue section. Your positive control refers to the epitope, so your regular positive control from a tissue section can be used, BUT you negative control has to be a thin prep from the same case simultaneously prepared.
  René J.

AWeiss@shorememorial.org wrote:
  
Is anyone doing IHC's on cytology specimens? If you are can you please
send staining and fixation protocol. Also, what do you use for your
positive and negative controls? Thank you

Andrea J Weiss BST CT (ASCP)
Cytotechnologist
609 653 3577 Ext 4907
aweiss@shorememorial.org
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