Eosin bleeding out

From:"Johnson, Teri"

I have this trouble using the xylene substitutes.  Most manufacturers say if you have absolutely anhydrous alcohol before your final clearing, this won't be a problem. I find that it's nearly impossible, especially in the humid conditions here, to keep the alcohol completely anhydrous. I'll get bleeding of the eosin after staining three slides, even with 4 changes of absolute reagent alcohol.

Having said that, I am evaluating Surgipath's Clearium, and am coverslipping directly out of absolute isopropyl alcohol.  It's too soon to tell if I like it unconditionally, but the bleeding problem seems to have stopped.  The only problem I've noticed with it is air bubbles appear several hours after coverslipping.  I suspect they were there all along, but just got bigger as they sat. 

Teri Johnson
Managing Director Histology Core Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110

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