Re: frozen mouse brain
Cynthia Favara wrote:
> I am requesting thought on long term storage of mouse brain. My current
> protocol is to take one half fresh or NBF fixed, sucrose protected mouse
> brain usually between 10 and 14 days. Surround with OCT freeze over liquid
> nitrogen and store in -80C.
What is meant by "freeze over liquid nitrogen?" For a specimen as
big as a whole brain of a mouse the only acceptable way to get
fast
freezing (essential if you don't want ice crystal holes) is to
immerse in a liquid - isopentane is the easiest one to use - that
has been pre-cooled to liquid N2 temperature. Thin slices can be
frozen on a very cold metal block, but a mouse's brain isn't a
thin slice. With cryoprotection, fixed objects as thick as 1 cm
can be frozen on a cold metal block well enough for most light
microscopy applications.
--
-------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
kiernan@uwo.ca
http://publish.uwo.ca/~jkiernan/
<< Previous Message | Next Message >>