Question on Snap Freezing

From:Stephanie Moore <>


Recently someone responded with a nice procedure on how they snap freeze
that seems like something I could do in my lab.  I have wanted to try
snap freezing for a variety of reasons, but did not have access to
liquid nitrogen, etc.  

The method I am interested in is the isopentane/dry ice slush.  What
kind of container should this be done in?  I am also assuming this needs
to take place in a fume hood (I like to know even the "obvious"
details). I am going to order the cryomolds from Sakura.  I just fill
the mold with OCT, place the tissue inside, orient it, then lower the
mold (being held with forceps or something?) into the slush for how
long?  I didn't see that these molds were peel-away...Is their removal
from the frozen OCT difficult (do I need scissors, razor blade, etc).

How do I then attach this OCT block to the cryostat chuck (I have Leica
chucks with the concentric ring grooves) and keep the specimen in the
same orientation?  

Is the isopentane reusable?

Thanks for the info -

Stephanie Moore 
Brandeis University
Waltham, Ma

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