Controls for fixation and Antibodies

Ross S. Chafetz rchafetz at cephalo.neusc.bcm.tmc.edu
Thu Aug 6 11:16:47 EST 1992


Thanks for the suggestions.

I did not realize the age of the tissue might be a factor.  I 
came across the problem when using younger
tissue - 12 days old.  This age, of course, is also
harder to perfuse.  With the possibility of fixation variance - are
there any controls commonly used to compare fixation from different tissues.

Here a little more of my proceedure

Fixation - 4% Para, intracardial
Post fixed - 12-48 hours
sucrose sink - 3-5 days
solution - All 0.1 M PBS (exept of the DAB reaction which is TBS)
storage - O.1 PBS with Sodium Azide


The paint brush is a good idea, but once the tissue sticks it would
not be enough to separate the sections.  One other suggestion I may
try is using a holder with a mesh bottom, making the transfer of solutions
easier.

I'm open to any other ideas, specially about fixation controls (no Freudian jokes
please :))

Ross 



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