Re: sectioning mouse eyes - help

From:Stephen.Eyres@sanofi-synthelabo.com


Hi,

We fix our mouse and rat eyes in Davidson's fixative, which, along with
ethanol, formaldehyde and methanol,  contains acetic acid. This helps
produce good lens and retina layer integrity. If soaking in water does not
help, try soaking in formic acid or Mollifix.

Cheers

Steve


                                                                                                                      
                    Jackie.O'Connor@                                                                                  
                    abbott.com              To:     "Caldas, Hugo"                 
                                            cc:     "'histonet@pathology.swmed.edu'"    
                    16/09/2002 12:19        Subject:     Re: sectioning mouse eyes - help                             
                                                                                                                      
                                                                                                                      





This may be a rhetorical question, but are you soaking your faced blocks on
ice prior to obtaining sections?  You won't be able to get more than about
6-8
sections per soak - but I've found this to work quite well for the 1,000's
of
eyes I've cut.

Jackie O'Connor
Abbott Laboratories
GPRD
Discovery Chemotheraputics
Jackie.O'Connor@abbott.com
847.938.4919



                    "Caldas, Hugo"
                    

                                                     cc:
                    09/14/2002 05:39 PM              Subject:
sectioning mouse eyes - help






I have been struggling for a while so I thought I would seek advice/help
from histonet.
I have formalin fixed paraffin embeded mouse eyes I am trying to section on
a Leica microtome at 4-5 microns. I seem to almost invariably get a "hole"
where the lens should be as I get further down the eye, it just seems to
burst open (I know the lens hardens the older the mouse, but there have to
be ways of doing this).
Does anyone have any tricks, tips, advice, anything? I would like to
eventually perform immunohistochemistry on some of the sections and expect
to detect mostly in the lens, hence my dispair.

Thanks

Hugo

Hugo Caldas
Columbus Children's Hospital
Columbus, OH












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