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snap-freezing fixated GFP tissue

Branwen Morgan b.morgan at garvan.unsw.edu.au
Mon Apr 12 22:48:36 EST 1999


>X-SILO-POPserver: bramor at gimr
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>Date: Tue, 13 Apr 1999 13:19:39 +1000
>To: b.morgan at garvan.unsw.edu.au (Branwen Morgan)
>From: Laura Kus <l.kus at garvan.unsw.edu.au>
>Subject: Info for Francisco Fernandez
>
>        Below is information in response to Francisco Fernandez's question
>of 23 March 1999
>
>        Immersion of a warm mouse brain into liquid nitrogen will almost
>certainly result in shattering of the tissue.  You should never drop any
>biological specimen bigger than a few mm into liquid nitrogen if you want
>to preserve morphology.  Shattering can be avoided if place the brain on a
>metal platform that is SLOWLY lower into the liquid nitrogen.  Starting
>with tissue that is cold may also help.
>
>        Have you tried to freeze your tissue on dry ice?  A mouse brain
>should freeze relatively quickly.  The tissue can be placed on foil that
>sits directly on a slab of dry ice.  As soon as the brain sticks, wrap the
>edges of the foil around the specimen and cover it with small pieces of
>crushed dry ice.  The brain should freeze within a minute.
>
>        Remember it is important to cryoprotect (sucrose or glycerol) after
>paraformaldehyde fixation if you want nice morphology.
>
>
>





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