[Histonet] Freezing Pig Eyes??

[Rebecca Jo]

Hello all,

I will be obtaining some pig eyes in the near future and would like to know if
anyone has a protocol for fixing and freezing pig eyes.  I will be using the
sections for in situ hybridization and immuno-staining.  I've been working
with frog eyes with some success but am finding it difficult to obtain really
great sections.  I usually make a slit in the eye, fix it in 4%
paraformaldehyde overnight, and then put it through a series of 10%, 20%, and
30% sucrose.  Then I snap freeze the tissue in isopentane cooled by liquid
nitrogen.  As stated before, I've had some success with this protocol but am
unsure as to whether this will work with pig eyes.  I've heard they can be
tricky, especially with the retina becoming detached during the freezing
process.  If anyone has any tips for me, I would greatly appreciate it!

Thanks.

-- 
Rebecca E. Jo
UNC-School of Medicine
Department of Cell and Developmental Biology
(919)843-9648
CB# 7090