RE: [Histonet] eosinophils on frozen section

From:"Weems, Joyce"

Fix in formal-alcohol (no, it is not wearing a tuxedo) 90 ml absolute
alcohol - 10 ml 37% formaldehyde... Should work ok then... j

-----Original Message-----
[] On Behalf Of Ingles
Sent: Tuesday, February 05, 2008 7:08 PM
Subject: RE: [Histonet] eosinophils on frozen section

My guess is that the alcohol is lysing(blowing up) the unfixed
eosinophils, therefore they are not there to stain. Fixation stabilizes
the cells in order to be stained in regular paraffin sections.
Unfortunately, I have done a few trials using different fixatives for
fresh tissue to see if I could preserve these little suckers. Nothing
really worked satisfactorally. You might try fixing in formalin for a
minute or two before you continue on with your stain and see if this
helps. (don't forget to wash the sections afterward) Hope this helps. 

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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