[Histonet] Frozen storage of tissue and sections,
what is the correct temperature?
Kelly D Mcqueeney
kelly.mcqueeney <@t> bms.com
Thu Feb 17 10:52:36 CST 2005
I know that frozen tissue storage is a common topic, but I have a hard
time finding the right answer throughout the histonet archives.
We are currently coating brain in embedding medium and snap freezing
brains in isopentane followed by immediate storage at -80C. Then, we
bring the brain up to -20C a few hours before sectioning. After
sectioning, we dry the slides at RT for several hours and store at -80C
until using the tissue for ligand binding, IHC or insitu. This exact
protocol has been moderately successful, so far so good. But, some of
the staff store frozen brains at -20C. They feel that keeping the tissue
at cryostat temperature reduces the amount of freeze artifact. I was
always taught to freeze fresh frozen brain at a much lower temperature.
None of this has been tested in our lab. Any suggestions for long-term
storage of whole brains after collection? Is -80C the correct
temperature? Will ice crystals spread at -20C? Will the receptor
affinity decrease or protein degrade more rapidly at -20C? Any feedback
would be greatly appreciated.
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