Re: [Histonet] cryoprotection

From:John Kiernan

   Transfer  the  brain,  after  adequate fixation in buffered
   forma=  ldehyde, to a 30% solution of sucrose in water. When the
   brain  sin=  ks  (1-2  days)  it is ready for trimming, freezing and
   cutting  sections.=   Cryoprotection isn't perfect. You
   should  still  freeze  as  q=  uickly  as  possible.  Put  the brain
   on a  metal  cryostat  chuck,=  with a drop of water or OCT
   compound  for  adhesion, then stand the chu= ck in a slurry of solid
   CO2  and  either acetone or alcohol. Mount the c= huck with specimen
   in   the   cryostat   and   wait  for  its  temperature =  to
   equilibrate. This isn't the fastest way to freeze thing    but  it's  good  enough  for cryoprotected formaldehyde-fixed rodent
   brain=   s.
John Kiernan
Anatomy, UWO
= London, Canada  
= = =
----- Or= iginal Message -----
From: Teisha Robertson <tshrobertso=>
Date: Monday, May 5, 2008 12:41<= BR>Subject: [Histonet] cryoprotection
To: histonet@li

> i am trying to cryopr= otect the mouse brain.  i place the
> brain in= 10% formalin,70%ETOH and 30% sucrose. can you please
= > tell me what i need to do to prevent holes in my sample?>       
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