[Histonet] E16 paraffin embedding

[Rene J Buesa]

If your problem is sectioning, probably the problem is due to infiltration and the cause of the defective infiltration could be fixation, dehydration and/or clearing.
Without knowing your protocol it is difficult to try to try to find out the cause.
René J.

--- On Fri, 1/9/09, James Dooley <jdooley2008 <@t> yahoo.com> wrote:

From: James Dooley <jdooley2008 <@t> yahoo.com>
Subject: [Histonet] E16 paraffin embedding
To: histonet <@t> lists.utsouthwestern.edu
Date: Friday, January 9, 2009, 12:06 PM

Hello All,

I
have been having trouble cutting whole embryo sections on E16 mice. I
think the problem is with the fixation process. I am currently using a
modified Carnoy's fixation (60% ETOH, 30% formaldehyde, 10% Glacial
acetic acid). I would like to use this fixation as I have optimized my
antibodies for this condition. The tissue is soft. I think it is due to
incomplete dehydration. Does anyone have a any suggestions for doing
tissue this large as I know over dehydrating can lead to problems also. 

Thank you,
James



      
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