re: processing rodent skin

From:"t.hacker@har.mrc.ac.uk" <T.Hacker@har.mrc.ac.uk>

Date sent:      	Wed, 10 Jan 2001 17:10:00 -0500
From:           	"Philopena, Jennifer" <jennifer.philopena@canji.com>
Subject:        	re: processing rodent skin
To:             	"'Histonet'" <histonet@pathology.swmed.edu>

> I am trying, without much success, to get mouse and rat skin processed so
> that the samples will section without blowing out or disintigrating in the
> water bath after sectioning.  I have a Shandon Citadel processor and have
> noticed that the problem improves when the processing reagents are fresh
> (that's a lot of reagent to change out!)  The skins are fixed for at least 2
> days in 10% formalin, and the mouse skin is processed on a 15 minute per
> reagent schedule (70%, 80%, 2x95%, 2x100%, 3xPropar, 2 wax baths).  The rat
> skin is processed on a 30 minute per reagent schedule.  The skin is kept
> flat in the cassettes using rectangular sponges.
> Any input would be so appreciated!

Jennifer,
you may find that increasing your processing times should improve 
matters. Despite being thin, rodent skin presents the usual 
processing problems,epidermis and dermis will separate readily if 
"under" processed. We attach the skin to card or cork with pins or 
staples, fix for 24-48 hours (this will harden tissue and prevent 
curling) then take slices for processing (standard overnight, approx 
1-2 hours in each reagent). Make sure your water bath for floating 
out sections is not too hot!
Terry.
Terry Hacker,
Medical Research Council,
Harwell,
Didcot,
Oxfordshire, OX11 ORD
01235 834393 x360



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