[Histonet] H&E staining

[Tony Henwood (SCHN)]

I would look at the dewaxing stage.

Have you changed waxes?
Try longer in xylene &/or longer in the oven.

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist, the Children's Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 


-----Original Message-----
From: Charles Riley via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Sent: Monday, 29 February 2016 10:16 PM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] H&E staining

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
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please delete it and notify the sender.

Views expressed in this message are those of the individual sender, and are not necessarily the views of NSW Health or any of its entities.