[Histonet] Freezing rodent brains for Mass Spec

[Gay Rudow]

We freeze mouse brains that have been perfused with 4% paraformaldehyde, put into PBS for 24 hours, and then into PBS with 30% sucrose overnight. We snap freeze them in methyl butane on dry ice. You have to put a thermometer into the methyl butane periodically to make sure that it doesn't get colder than -20 C or the brain will crack. Will the sucrose get in the way of what you are doing, or will this work for you?

Gay Rudow 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jackie M O'Connor
Sent: Tuesday, April 14, 2009 6:54 PM
To: histonet <@t> lists.utsouthwestern.edu; histonet-bounces <@t> lists.utsouthwestern.edu
Subject: [Histonet] Freezing rodent brains for Mass Spec

I have a colleague who requires snap frozen  rodent brains to perform Mass 
Spec on 10micron frozen sections mounted on plates.     Because OCT 
interferes with mass spec, they are currently using ethanol to slow the 
freezing process to prevent cracking of the brain in liquid nitrogen. The 
alcohol, however, causes some detriment to the surface of the brains - in 
science-speak "they get mushy".    Does anyone have any suggestions for 
snap freezing brains without any substrate while preventing cracking? 

Jackie O'

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