[Histonet] eosinophils on frozen section
Rene J Buesa
rjbuesa <@t> yahoo.com
Wed Feb 6 08:11:04 CST 2008
Use a counterstain made of: Eosin Y---0.8 g + phloxine---0.2g in 95% ethanol. Stain 1 minute and dip quickly in distilled water and the RUN (I mean RUNNING) through the alcohols. You can start in 100% EthOL, clear and mount.
René J.
"Breisch, Eric" <ebreisch <@t> rchsd.org> wrote:
To all interested Histonetters:
We are experiencing some frustration with our ability to identify
eosinophils from frozen sections. For some reason which our frozen
section set up does not enable us to identify eosinophils. Eosinophil
identification is not impaired when the tissue is submitted for
permanents. Could some one please share the reagents used and how the
frozen section staining area is set up if they have success identifying
eosinophils from a frozen section? We have had many frozen sections done
on fresh tissues suspicious for an eosinophilic granuloma but our
techniques so far have not rendered consistent eosinophil
identification. We presently are using 95% ETOH for fixing the FS slide,
quick wash in distilled H2O, 1 minute in Richard Allen Hematoxylin 2,
rinse in tap water, 15 seconds in bluing, rinse in water, rinse in 70%
ETOH, 1 minute in Richard Allen Eosin Y, three rinses in 95% ETOH, three
rinses in Xylene substitute and then coverslip. All other tissues appear
just fine but the eosinophils just don't show up. Any suggestions are
greatly appreciated.
Eric A. Breisch, Ph.D.
Clinical Anatomist
Dept. of Pathology
Rady Children's Hospital and Health Center
Associate Clinical Professor of Anatomy
Dept. of Surgery
UCSD School of Medicine
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