Re: Coverslips/Gaskets for ISH

From:Mikael Niku

Dear Kimberly,

there is no need for special coverslips etc. Just use
standard coverslips, wash them quickly in 100 % ethanol,
air dry completely, and use.

I'm applying the hybridization solution to the slides
first (about 100 ul is enough for a standard slide
with several sections), then cover it with standard
coverslip, denature for about 8 min in 85 C, pack
immediately in airtight containers, with a piece of tissue
paper moistened with basic ingredients of the hyb buffer.

I use hybridization times up to 2 days and there are
no problems. Some hyb solution is evidently evaporating
during denaturation (take care of the formamide fumes!),
but not enough to cause problems.

   Mikael Niku             URL:
   University of Helsinki  Dept. Basic Veterinary Sciences
       - Mitäkö mieltä olen länsimaisesta sivistyksestä?
         Minusta se olisi erinomainen ajatus!
                                              - Gandhi
----- Original Message -----
From: "Kimberly L Merriam" 
To: "Histonet" 
Sent: Wednesday, January 23, 2002 11:45 PM
Subject: Coverslips/Gaskets for ISH

> Hi,
> I will be starting some in situ hybridization (DNA probe) in
> the near future and am looking for some advice on types of
> coverslips and/or gaskets to use while incubating with the
> probe.  EMS has several different types (Probe-Clip,
> CoverWell, Hybriwell, Hybrislip, Perfusion Chambers and
> Press-to-seal Silicone Isolators).  Any opinions,
> advantages, disadvantages on them would be appreciated.  I
> don't know which one to try!
> Thanks in advance,
> Kim Merriam
> Cambridge, MA

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