staining old tissue

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From:Becky LoVerso <loverso-toth.1@osu.edu> (by way of histonet)
To:histonet@histosearch.com
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Hi.  I've got some mouse spinal cord tissue that has been sitting on slides
in a freezer for two years.  I need to stain it for oligodendricytes with a
cc-1 antibody.  The procedure that was prepared workes beautifully on
tissue that has been UNCUT in a freezer for a long time, and was cut only
recently to practice for this stain.  However, when the precut old tissue
(the important stuff) is used with the same procedure, the background
staining is terribly high.  Oh yeah, it's developed with DAB.  The
oligodendricytes are too light to be seen clearly against the background
stain, making them uncountable.  Our head histotech thinks its due to
freezer burn, or something similar to it. Does anyone have any ideas as to
how I can salvage the rest of this tissue?

Thanks,
Rebecca LoVerso
Dept. of Neuroscience
The Ohio State University




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