[Histonet] PEN SLIDES FOR LASER CAPTURE

[Amos Brooks]

Hi,
     I do sectioning on LCM slides frequently. The problem of tissue coming
off the slides si kinda the whole point of the LCM slides. The tissue is
supposed to come off, otherwise it would be impossible to capture the tissue
with the laser. This of course poses a problem for staining. First you need
to figure out what you are extracting. If it is for PCR for DNA you are in
luck, RNA causes more complications because of the RNAases that can destroy
the yield.
     You will not be able to stain the tissue the same way you would for
regular charged or adhesive slides. You can heat the slides some, but bear
in mind this will increase RNAase activity. Xylene will cause the plastic
membrane to shrivel and fall from the slide, so you will need to abbreviate
the deparaffinization to only a couple of minutes. Yes this will probably
cause the staining to be less than perfect, but perfect staining is not
really needed on LCM slides (If you want perfection use charged slides).
Rehydrate quickly and gently, shorten the time in the stains as best as you
can.

Best of Luck,
Amos



Message: 14
Date: Thu, 5 Aug 2010 12:51:48 -0400
From: "Mary Lloyd" <lloyd.3 <@t> osu.edu>
Subject: [Histonet] PEN SLIDES FOR LASER CAPTURE
To: <histonet <@t> lists.utsouthwestern.edu>
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Does anyone have any experience with PEN membrane slides from Zeiss for
laser capture microdissection.  I have been working with one of our
residents and we are having problems with the tissue staying on these
slides. We could sure use help.  Thanks Mary Lloyd