Re: sucrose/OCT

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From:Geoff McAuliffe <mcauliff@UMDNJ.EDU> (by way of histonet)
Date:Mon, 31 Jan 2000 22:47:09 -0500
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denise M m Long-Woodward wrote:

> Hi! I'm looking for instructions on how to "cryopreserve" mouse tissue
> using sucrose and OCT. Details will be appreciated.  Many thanks for your
> insight.


    After fixation and several buffer washes, I soak the tissue in cold 10%
sucrose in buffer until it sinks,usually 1-3 hours depending on size. Then
cold 20% sucrose in buffer until it sinks which is usually overnight. I have
used concentrations of sucrose as high as 30% but see no benefit. I only  use
OCT to "glue" the tissue to the microtome stage.
    Rapid freezing of the tissue is very important to prevent ice crystal

Good luck,
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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