Hi Holly,
 
you should be able to leach out the eosin with alchohol. I am not sure that your antigens are going to be very healthy after various treatments with alcohol, high and low pH buffers etc.
 
The other choice would be to bleach the eosin out with your mercury lamp. So it will still be there, just not fluorescent.
 
 
Good luck
 
 
Cam
 
 
 
Cameron J. Nowell
Microscpy Manager
Central Resource for Advanced Microscopy
Ludwig Insttue for Cancer Research
PO Box 2008
Royal Melbourne Hospital
Victoria, 3050
AUSTRALIA
 
Office: +61 3 9341 3155
Mobile: +61422882700
Fax: +61 3 9341 3104
 
http://www.ludwig.edu.au/branch/research/platform/microscopy.htm
 

________________________________

From: Confocal Microscopy List on behalf of Holly Aaron
Sent: Fri 31/07/2009 7:26 AM
To: [log in to unmask]
Subject: releasing old histology stains (which are fluorescing in red)


Dear List - Not strictly a confocal question, but...

Does anyone have a protocol for releasing old histology stains from parafin embedded tissue sections? I have users who are having a hard time getting around a lot of red background in their samples. They have already removed the parafin and have been able to release some of the stains using high or low pH baths, but still see a significant signal in the red channel. They want to do multi-color antibody labeling on the sections now. They can do it avoiding red, using green and blue and maybe a far-red (we have not characterized the spectra yet), but if there is a way to release these old stains, it would be very useful for them. They think the only stains on the tissue are H&E (which they think is not the problem as they have done this before, although eosin is brightly red?) and PAS-aurantia. 

Thanks for any tips or ideas!


-- 


Holly L Aaron

Molecular Imaging Center

Cancer Research Laboratory

251 Life Sciences Addition #2751

Berkeley, CA  94720-2751

510.642.2901

510.642.5741 FAX

[log in to unmask]

http://imaging.berkeley.edu <http://imaging.berkeley.edu/>