isopentane freezing of rat brain

Hector Gonzalez hgpardo at uniovi.es
Thu Mar 3 11:05:53 EST 2005


I am freezing rat brains using isopentane cooled at - 40 ºC (the
isopentane bottle is chilled in an ultrafreezer). Usually, I lower the
brain slowly into the chilled isopentane during aprox. 30 secs in a
Pyrex glass vase placed in a freezer at -20 ºC. The brain remains
there for about another 30 secs and then I store it in a freezer at
-40ºC. Using a cryostate, I obtain 40 micrometer-thick sections at
-20ºC. However, sections look good, but under the microscope seem to
be a bit micro-cracked or granulated. I would greatly appreciate any
tips or suggestions to improve the quality of the brain tissue.



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