Frozen tissue problem

From:Connie McManus <>


I have an interesting problem with that frozen mouse liver I am working
with.  Not long ago, I asked for, and received wonderful responses,
about the best way to fix frozen tissue.  Well, we tried defrosting the
tissues in cold 10% BNF, but what we got was what appears to be staining
on the periphery of the tissues (the Dr thinks it is staining, I have my
doubts) and nothing in the interior.  Worse yet, some of the tissues
look like they have bad freezing artifact.  I have never actually seen
freeze artifact before, so I'm only guessing, but the cells are
irregular, badly stained, vacuolated, and seem to be held together by a
network of strings.  In short, they look tattered and torn.

The fellow who does this research wants to try antigen retrieval on
these to see if we can salvage anything.  The big question is this...
We're working with hepatitis B and  the antigen is inside the cells.  If
we have ruptured the cells, wouldn't all the antigenicity of the cells
have been washed out in the processing??   Does anyone have any
suggestions to help out?  Unfortunately ALL the tissue in this study was
frozen in LN2  with out cryopreservatives, so we are desperate for help
with this.  I deeply appreciate your comments.

Connie McManus

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