Our procedure for cytospins & smears intended for
IHC or/and ISH (in situ hybridization):

- air dry (at least a few hours after preparing)
- fix in 100% ethanol at -20C for 30 min OR
  in acetone at -20C for 10 min
- air dry (at least 30 min)
- store at -80C in small airtight containers (5 slides each)

We don't use nuclear antigens currently but as genomic ISH
works there seems to be no permeability problems.

We switched to storage at -80C after problems with samples
stored at -20C.

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   Mikael Niku             URL: www.helsinki.fi/~mniku/
   University of Helsinki  Dept. Basic Veterinary Sciences
       - Mitäkö mieltä olen länsimaisesta sivistyksestä?
         Minusta se olisi erinomainen ajatus!
                                              - Gandhi
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----- Original Message -----
From: "David Grehan" 
To: "'histonet'" 
Sent: Thursday, October 25, 2001 1:49 AM
Subject: fixation procedures for cytology immunocytochem


> Hi histocybernetters
> I would like to get your opinions on procedures for cytology specimens
> that are prepared for  immunocytochemistry. I am currently fixing direct
> smears or cytospins immediately in 95% absolute alcohol for 10 mins .I
> do this for any cytoplasmic or cell membrane antigens . I fix in
> methanol:acetone 50:50 for 10 mins for any nuclear antigens such as Tdt
> or Myo D1 (to increase permeabilisation)is this necessary?. I work in a
> busy paediatric laboratory so we don't get a lot of cytology specimens
> but on occasion we are requested to do them. My results to date have
> been patchy especially following storage at -20C. I would like to be a
> bit more confident about my procedures and would like  any comments from
> anyone doing a lot of immunocytochemistry on cytologyy specimens . Also
> how do you store your smears ? Do you spray fix or use PEG.?
>
>
>