Re: insitu on blood smears

From:Mikael Niku <>

Dear Betty,

we are routinely performing in situ hybridization on
cytospin preparations and occassionally also on smears.

Either way, SuperFrost slides are used. Cytospins /
smears are let dry at room temperature (usually from
a few hours to overnight), then fixed either in 100% ethanol
at -20C for 30 min or in acetone at -20C for 10 min,
let dry at room temperature, and subjected to in situ
hybridization or stored at -80C.

I don't recall any significant problems with samples
dropping off the slides. Our ISH protocol is pretty harsh,
and in order to acquire best possible morphology,
the slides are briefly fixed in 4% PFA during the procedure.

   Mikael Niku             URL:
   University of Helsinki  Dept. Basic Veterinary Sciences
       - Mitäkö mieltä olen länsimaisesta sivistyksestä?
         Minusta se olisi erinomainen ajatus!
                                              - Gandhi

----- Original Message -----
From: "Gaskill, Betty A" <>
To: <>
Sent: Friday, July 20, 2001 1:00 AM
Subject: insitu on blood smears

> Hello,
> Anyone had experience with insitu on blood smears?  Any info on specimen
> prep and fixation would be appreciated.  We have had trouble with the
> coming off the + charge slides immediately when placed in fixative.  The
> smear is done by a machine not by hand, air dried for 15 minutes and the
> into 4% paraformaldehyde.  Immediate fixation with no air drying has also
> been tried and the prep still falls off.  I have suggested a few  things
> them to try, but thought I would tap into histoland's vast experience.
> Thanks for your help.
> Betty Gaskill
> Safety Assessment
> Molecular Pathology
> GlaxoSmith

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