> Date:          Tue, 26 Jan 1999 11:41:55 -0600 (CST)
> From:          Carolyn Pressman <cpress@gsbs3.gs.uth.tmc.edu>
> Subject:       Fixation and embedding of adult murine eyes
> To:            HistoNet Server <HistoNet@Pathology.swmed.edu>

> 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?

Hi Carolyn,
Eye lens is a nightmare to preserve, I have tried many techniques
some successful, others disasterous. For paraffin wax, 24 hour
formalin fixation followed by 48 hours in Davidsons (30ml  95%
alcohol, 20ml buffered formalin, 10ml glacial acetic, 30ml distilled
water ), followed by a lengthy process over 2 days to wax. This seems
to give good preservation of both retina and lens.
The trick seems to be in the cutting, even after this process.
Trim to the lens, soak the cut surface in 1% phenol ( on tissue paper
in a dish for 10-20 mins, cool on ice, cut immediately keeping the
block face moist. This should give you a ribbon of sections before
the lens becomes dry and cracked. Repeat the procedure for the next
sections.
A bit tedious but this does give a nearly intact lens ( except for
the centre).
All of this was done on adult mouse eyes looking for cataract
development.
Good luck,
Terry.
Terry Hacker,
Medical Research Council,
Harwell,
Oxfordshire,
U.K.