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.

Denise:

    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
artifacts!

Good luck,
Geoff
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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
mcauliff@umdnj.edu
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