......."I find that it's nearly impossible, especially in the humid
conditions here, to keep the alcohol completely anhydrous."

Teri,

have you considered the use of DRIERITE (anhydrous calcium sulfate) in your
absolute alcohols.  the color indicator will give you feedback on how
relatively absolute your absolute really is.

rob

-----Original Message-----
From: Johnson, Teri [mailto:TJJ@Stowers-Institute.org]
Sent: Friday, August 09, 2002 11:48 AM
To: Histonet (E-mail)
Subject: Eosin bleeding out


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
tjj@stowers-institute.org


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