Just a few ideas:
Do not cryoprotect unfixed tissue, it will just degrade.
Do not immerse the brain directly in isopentane. Remove brain from pup,
mount with a dab of OCT on chuck, put stem of chuck in -150 isopentane
cooled with liquid N2, brain will freeze in 30-60 seconds.If your chucks
are
Dear Histoneters,
I am looking for help regarding flash freezing of rat pup brains
(Postnatal day 8). We need the tissue to be fresh (unfixed) for
cutting at the cryostat.
So far the technique has been:
1- to scoop the brains straight into cold (-50 C) Isopentane for
about 10 seconds,
2
