Re: IHC storage alcohol
|From:||Karen Larison <email@example.com>|
In a pinch, we have stored fixed whole mount embryos in methanol kept
at -20C. You must first dehydrate. I think this yields high
background, but it's better than nothing.
Also, we have stored fixed mouse organs at -20C in the following
500 mL 0.1 phosphate, pH 7.2
300 g sucrose
300 mL ethylene glycol
bring volume to 1 liter with distilled water.
I then wash them in several changes of 30% sucrose before
cryosectioning. It seems like it would work for tissues fated for
paraffin sectioning after washing thoroughly with water. The
original reference on this technique: Use of Cryoprotectant to
Maintain Long-Term Peptide Immunoreactivity.." Peptides 7. pp
Karen in Oregon
>Usually the tissue samples I receive have been optimally fixed (time-wise)
>in formalin and promptly embedded into paraffin. Today I received a call
>from a researcher out of state who wants to send me some samples, but cannot
>get his fixed tissue processed into paraffin within a 24-hour period after
>start of fixation. He wants to know what storage alcohol he should switch
>the samples into until embedding, and I became brain-dead. Can someone help
>me out here PDQ? Is it 70% ethanol or some other percentage? Is there a
>better storage medium than ethanol? I have received some cases in PBS
>(post-formalin-fixation), but those staining results were less than optimal.
>Any help would be greatly appreciated.
>Univ. of Minn. Vet. Diag. Lab
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