[Histonet] Frozen Section H&E's

Atoska Gentry gentras <@t> vetmed.auburn.edu
Wed Jan 24 10:21:45 CST 2007

Hello, I use the following H&E protocol for frozen sections:

1. Stain in Mayer's hematoxylin 5min.
2. Wash in tap H2O 5min.
3. 1% Eosin 3min.
4. Wash in deionized or distilled H2O 3min.
5. Dehydrate in ascending ethanols (ETOH): (x2) 95%,  (x2) 100% all 2 
min. each (ea.)
6. Clear in (x2)  Hemo-de (xylene substitute) 2 min. ea. then xylene
7. Mount in mounting medium of your choice ( I use cryoseal -60)

             _1% Eosin
_Eosin Y 5g.
Distilled H2O 750 ml
2% Acetic Acid 10 ml.
100% ETOH 250 ml.
Dissolve Eosin in H2O add alcohol & acid. Ready for immediate use.
Best wishes, Atoska

LeVier, Rebecca J wrote:
> Hi Folks, 
> I need some help, please.
> Would someone be willing to share reagents and times of the H&E stain
> for frozen sections if you are using a xylene substitute?
> We are having a hard time clearing....
> Thanks so much in advance.
> Becky
> Rebecca J. LeVier BS HT(ASCP)
> Histology Supervisor
> Email: rjlevier <@t> lancastergeneral.org
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Atoska S. Gentry, B.S., HT(ASCP)
Research Assistant IV
Scott-Ritchey RSCH Center
College of Vet. Med
Auburn, AL 36849
PH (334) 844-5579
FAX (334) 844-5850
email: gentras <@t> vetmed.auburn.edu

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