[Histonet] removing cryoprotectant from fixed tissue
Karl Garsha
garsha <@t> itg.uiuc.edu
Tue Mar 4 15:39:07 CST 2008
Greetings,
I'm trying to help someone who has some formalin-fixed tissue sections
that have been cryoprotected in a 30% ethylene glycol/20% glycerol
solution; we'd like to process the tissue for laser microdissection for
purposes of RNA extraction. The difficulty is that cryoprotectant tends
to absorb the laser wavelengths and heat the tissue during the cutting
process, causing damage and decreasing the liklihood of successful RNA
extraction (the likelihood of which has already been reduced by the
fixation process).
I would imagine that there must be a way to de-cryoprotect through
successive washes in a miscible solvent, I'm not sure if this would
cause even greater complications with RNA work, or if fairly gentle
standard protocols exist so I'm appealing to the collective knowledge of
this list:) Has anyone experienced a need to remove cryoprotectant from
tissue (brain) sections and are there perhaps methods to accomplish this
that might be somewhat amenable to later extraction of RNA from select
features in the tissue sections?
Thanks in advance for any advice or helpful hints.
Sincere Regards,
Karl
--
Karl Garsha
Research Microscopy Specialist
US-Southwest Region
Leica Microsystems-Life Sciences Research
www.leica-microsystems.com
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