Re: Fixation and embedding of adult murine eyes

<< Previous Message | Next Message >>
From:Lynn Gardner <> (by way of histonet)
To:histonet <>
Content-Type:text/plain; charset="us-ascii"

Dear Carolyn,

You came to the right person, mostly what we process is eye tissue from
human to murine. We find that Pen-fix which is an alcoholic formalin from
Richard Alan (1-800-781-2307) works very well. I would be happy to send you
two protocols we use for processing these eyes that work very nicely. One
is an overnight process and as long as the tissue has fixed for 4 hours
prior the tissue will do fine. We also have a very quick process which we
can do. The tissue needs to fix for 4-6 hours and then we run a short cycle
on the processor the difference is how fast the researcher needs their
materials. We perform routine, special and immunohistochemistry on our
tissues and this fixative seems to enhance the staining qualities of the
tissues and is very versitile.

As for the lens problem, we have found that if you take a 0.25-0.5%
ammonium hydroxide (in water) and pour it into a dish containing ice
carefully trim you block so that the lens is exposed. Place the block into
the ice/Ammonium hydroxide solution and leave it sit for 15 minutes. Then
try cutting the tissue this solution will usually hydrate the tissue enough
to allow the lens to cut beautifully, if not soak the block a little
longer. If this does not resolve the problem try backing down the times in
your paraffins, xylenes and 100% alcohols during processing decrease them
each by 10 minutes to start and see if this helps the situation.

If you have any futher need of eye or other information please do not
hessitate to let me know. Good Luck!

Lynn Gardner, HT(ASCP)
Supervisory Research Assistant III
FC Blodi Eye Pathology Laboratory
The University of Iowa
233 MRC
Iowa City, IA 52242-1182
Phone: 319-335-7095
Fax: 319-335-7193

At 11:41 AM 1/26/99 -0600, you wrote:
>What is the best fixative to use for adult murine eyes?  Does anyone have
>any good protocals?  I have found through trial and error that fixing in
>neutral buffered formalin only works ok if I fix for 2 days.  I embed in
>parafin and then section at 7 um.  Most of the architecture of the eye is
>intact, but the lens always tears or has holes in it.  Is there a way to
>get around this problem?  Is embedding in plastic a better way to do this?
>Carolyn Pressman
>MD Anderson Cancer Center
>University of Texas--Houston

<< Previous Message | Next Message >>