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=  n@yahoo.com>
Date: Monday,
   May    5,    2008    12:41<=    BR>Subject:   [Histonet]
   cryoprotection
To: histonet@li   sts.utsouthwestern.edu

>   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?>
            
>  ---------------------------------
> Be
   a=   better   friend,   newshound,  and  know-it-all  with  Yahoo!
   
>=  ;  Mobile.  Try it now.
>
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