Re: EYES; frozen and paraffin

From:Roger Moretz

I was just perusing some back issues of J
Histotechnology, and found an excellent reference on
canine eyes, v. 14, no. 4, December, 1991, pp. 251-5,
J. A. Saby et al.  
From personal experience.  I think that Zenker's is my
preferred fixative for routine histology.  If you are
doing whole eyes (not enucleated), and you want to
avoid shattering of the lens, microtomy technique is
important.  We have been facing very slowly, chilling
the blocks during sectioning, and even soaking
throughout, using glycerol in the bath.  The problem
here is that the pathologists' preference for fixation
is 3% glutaraldehyde (unbuffered), and that can make
for little rocks (the lenses).  So, for small animals
(rodents), we fix for about 24hrs, wash, immerse in
NBF and embed immediately.  Large animals (canine,
etc), fix for 48hrs, trim, immerse in NBF and embed
immediately.  This protocol results in excellent
fixation, good preservation of the lens (without
shattering) and (generally) intact retina/rpe (in
fact, I think most of the artifact we get there is due
to necropsy technique).
Hope this helps.
--- Kristin Gebhard Mitchell
> Hi all...first time posting...
> Would those of you who have experience with fixing,
> embedding or cutting 
> eyes (mouse, rat, rabbit) be willing to contact me
> to discuss optimal 
> protocols? I've searched the archives, which were
> helpful, but I've got 
> specific issues.
> thanks much!!
> Kris
> -- 
> Kristin Gebhard Mitchell
> Histology Core Lab
> Department of Ophthalmology
> University of Minnesota
> 370 Lions Research Building
> 2001 6th St SE   Mpls, MN 55455
> 612-624-8542

Do You Yahoo!?
Send your FREE holiday greetings online!

<< Previous Message | Next Message >>