[Histonet] H&E staining

Charles Riley criley at dpspa.com
Mon Feb 29 05:15:54 CST 2016


Hello all,

 We've been running into an issue lately with our H&E staining. The
hematoxylin and eosin are light on and off but the chromatin is
consistently staining light. I have changed the time in hematoxylin to
longer, used a fresh lot of both hematoxylin and our clarifier solution and
the stain is not improving.  Does anyone have any suggestions?

Here is our routine stain line procedure.
65 degree Celsius oven for 20 minutes
2x xylene for 5 mins
2x 100% alcohol for  1 minute
95% alcohol for 1 minute
DI water rinse for 1 minute
Richard Allan scientific hematoxylin for 4 minutes
DI water rinse for 2 minutes
Richard Allan Clarifier 1 for 2.5 minutes
DI water for 2 minutes
Bluing Reagent for 1 minute
DI water rinse for 2 minutes
95% alcohol for 1 minute
Eosin Y for 2 minutes
95% alcohol for 30 seconds
2x 100% alcohol for 1 minute each
2x xylene for 1 minute each
Mount and coverslip.
-- 

Charles Riley HT(ASCP)CM

Histopathology Coordinator/ Mohs

Doctors Pathology Services, Dover DE


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