I am answering this from a research point of view, totally unaware of
Her2neu guidelines or what so ever.
We fixed cytospins in NBF (5 min, RT), washed them with tap water (5
min) and then to alcohol 70 (3 hours - overnight). Wash with tap water
and perform HIER with appropriate buffer. We use half the time at max.
temperature as for FFPE's. After cooling down, wash with tap water and
perform IHC as with FFPE tissue sections (antibody, dilution,
I am aware this procedure as against all logics but it simple
works. After all, 5 min NBF fixation cannot cause any cross linking
and alcohol fixation certainly not. Therefore, HIER would not be
necessary. But after testing we saw HIER was very essential! When I
heard about this procedure I was very sceptical it would ever
work. However, I was surprised to see how well the cells survive this
harsh procedure and nicely stay at the glass. For some particular
antigens we have even done this procedure successfully with cryostat
tissue sections from fresh frozen blocks (section stays at the glass,
morphology is fine)!
Therefore folks: a theory is good to keep in mind, but testing is
sometimes better. And, I also stopped with surprising myself about
I hope this helps.
Have a nice weekend,
Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
NL-1105 AZ Amsterdam
Date: Thu, 24 May 2007 11:16:19 -0500
From: "Deathridge, Mary Ann"
Subject: [Histonet] IHC cytology specimens
Does anyone in histoland have a protocol for the prep of cytology
specimens (cytospins, smears) for IHC?
Formalin fix? how long? air dried? 95% alc fix? esp. since the new
Thanks in advance
MaryAnn Deathridge, HT (ASCP)
Vanderbilt Un. Med. Ctr
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