[Histonet] Immunocytochemistry on cytospins

[Glencross Hedley (RW3) CM&MC Manchester]

Hi Gudrun

ICC on cytospins is easier if you follow a few fundamentals. 

Fresh samples are best, and a monolayer must be achieved. This requires
experimentation, but with experience the dilution factors and spinning
times can be found. Air dried samples are OK, with post fixation in cold
acetone, but if you decide to fix then as suggested light fixation is
all that is needed, although I have kept samples fixed up to 4 days
without too much problem. However, for suspected lymphomas air dried
cytospins are best. Lastly, if you do a peroxidase method this will need
a long block (around 30 minutes) in both hydrogen peroxide and non
immune serum to reduce background.

Have fun

Good luck

Hedley Glencross
Manchester Cytology Centre



Message: 1
Date: Tue, 8 Mar 2005 10:36:01 -0600
From: "GUTIERREZ, JUAN" <juan.gutierrez <@t> christushealth.org>
Subject: RE: [Histonet] cellsmears and cytospin for ihc
To: <gu.lang <@t> gmx.at>,	"Histonetliste (Histonetliste)"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	
<A61F23E937E4DA488E0C0F60093843D901E49834 <@t> ccetxm030.echristus.net>
Content-Type: text/plain;	charset="iso-8859-1"

We do a few markers regularly on cytospins and or smears.  The general
rule is to fix in 95% alcohol right after spinning, do not let them
air-dry.  From the alcohol we then transfer to the immuno buffer and go
from there.  We have had good results with storing the cytospins in the
same alcohol for short periods of time, but I wouldn't try it for very
long.  Good luck.

Juan C. Gutierrez, HT(ASCP)
Histology Laboratory Supervisor
(210)704-2533

My opinions are my own and do not reflect those of my employer.  Long
live free speech!


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gudrun
Lang
Sent: Friday, March 04, 2005 10:54 AM
To: Histonetliste (Histonetliste)
Subject: [Histonet] cellsmears and cytospin for ihc

Hi histonetters,

We perform ihc on cellsmears and cytospins very rarely. Therefore we
have no
really experience. 

Can anyone tell me, if there is a general preperation protocol for these
specimen, that is best for ihc?

Air-drying / fixation in . / storage ?

 

Or does it depend on the antibody or the kind of fluid?

 

Thanks in advance

 

Gudrun Lang

Linz, Austria