This is in response to a rather old message from Linda Graeter, about making
vibratome sections of fresh rodent and human skin.  (Sorry so late but I was
on vacation).  I didn't see any responses from people who had actually tried
skin, which is too bad because I would like some tips myself.

In the past I have had great success on fixed organs such as murine spleen
and lung using a method taught to me by Richard Burry (J. Histochem.
Cytochem. 40(12):pp.1849-1856, 1992.)  This method involves fixing the
tissue in paraformaldehyde, then mounting the tissue block onto the metal
cube provided by the vibrating microtome supplier (in my case EMS) by means
of cyanoacrylate glue.  (Instead of using the metal mounting blocks one can
also cut a piece of glass to the same area, so it can be held by the vise.)
The "super glue" must be very fast-drying, as in instant, or the tissue will
dry out.  Then drop some low-temperature agarose (0.7%, "Sea Plaque" or
similar) over the tissue to provide extra support.  Section with the
reservoir filled with buffer.  With this method I got fairly decent 100 -
200 micron sections.

I have also had very good success using this method to section FIXED pig
skin, although I found skin slightly more difficult than lung and spleen.
However, when I tried it on FRESH human skin, it was a disaster.  The
flexible fresh tissue just bent or squished but wouldn't cut, and the
agarose did not provide enough support.

So, I would also welcome some suggestions on how to do fresh (not frozen)
tissue!

Thanks,

Karen

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3M Company
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These opinions are my own and may not represent those of 3M.
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