RE: Re Oil red staining
|From:||"Saby, Joseph" <Joseph.Saby@pfizer.com>|
Whenever I've worked with those plastics, there has always been a clearing
stage through acetone. Since acetone would remove all non-bound fat, Oil
Red O would have nothing to go into.
When I've worked with these plastics, I also did a post-fixation in osmium
tetroxide, which does a very good job of staining fats and lipid (it turns
them black). Perhaps this would work for your purposes.
Joseph A. Saby, BA, HT(ASCP)
Drug Safety Evaluation
Pfizer Global Research and Development
2800 Plymouth Road
Ann Arbor, MI 48105
From: RichardWHorobin@aol.com [mailto:RichardWHorobin@aol.com]
Sent: Thursday, May 17, 2001 2:49 AM
Subject: Re Oil red staining
Has anyone used Oil Red O to stain lipids in tissues embedded in plastics
(Epon or Epon-Araldite)? If so, has this been done by staining en bloc or
by staining the sections. Sections would range from 1-4 microns in
A couple of people so far have commented on how hard this would be, and our
experience agrees with theirs.
HOWEVER Jaclynn also said:
We would also consider tissues embedded in glycol methacrylate. We'd like
to avoid frozen sections because we'd prefer the higher level of detail
possible with plastic.
In THAT case things look better! Would you settle for Sudan black, rather
than Oil red staining of the lipid? If 'yes' then there is a method - which
does indeed show even tiny droplets of lipid very clearly. This was worked
out by the one-time king of GMA staining Peter Gerrits, and can be found in
Neurosci Methods, as follows:
Gerrits PO, Brekelmans-Bartels M, Mast L, 's-GrAavenmade EJ, Horobin RW
and Holstege G. (1992)..
Staining myelin and myelin-like degradation products in the spinal
cords of chronic experimental
allergic encephalomyelitis (Cr-EAE) rats using Sudan Black B staining
of glycol methacrylate-embedded
J. Neuroscience Methods. 45, 99-105
Bye - Richard Horobin
Institute of Biomedical & Life Sciences, University of Glasgow
T direct 01796-474 480 --- E RichardWHorobin@aol.com
"What should we expect? Everything."
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