[Histonet] removing a coverslip

[Mindy Johnson]

Yes it takes a long time, but I too have found this is what works best.  In
fact I have had to restain several hundred slides here and that is what I
have done.  If it's a few slides, I would change the xylene a few times
cause I ruined a few when I didn't.  The mountant wasn't all removed.
However, I haven't had a problem with the prolonged xylene exposure and
restaining.  Good luck!  :)

 Mindy A Johnson
SRA II
UCSD - School of Medicine
Medical Teaching Labs

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Thursday, October 18, 2007 2:20 PM
To: N Fournier; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] removing a coverslip

The recommendation you received is the best.
  René J.

N Fournier <nfournier <@t> sasktel.net> wrote:
  Hi everyone, 

I need to remove a coverslip from tissue that was stained previously for
GAD67, so that I can counterstain the tissue with cresyl violet but also
retain the previous immunostain. What is the best method to remove a
coverslip ? The slides are Superfrost Plus, the tissue is rat brain, and
mounting medium is DPX or Entellan (depending on experiment). One
recommendation I received was to leave the slides in xylene for several days
and try to pry of the tissue that way. I would think that prolonged xylene
exposure might influence the staining of my tissue and secondly it seems to
be taking forever. 

If the xylene method is the best then I do not mind waiting, but I really
need to get this tissue counterstained ASAP. So any recomendations are
welcomed. And lastly, once the coverslip has been removed do I need to run
the slides through descending alcohols and water before I start my cresyl
violet staining protocol? If this is required, does anyone have handy the
concentrations of alcohol and how long the slides are to be kept in these
solutions? 

I really appreciate the help,

Neil


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