eosin spots on slides

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From:"McCollough, Carol" <CMCCOLLOUGH@dnr.state.md.us>
To:'histonet' <histonet@pathology.swmed.edu>
Content-Type:text/plain; charset="iso-8859-1"

Hi Histonetters:

I have a problem with our eosin Y-phloxine B leaving precipitate rings on
the bottom end of our H&E slides, usually on the back of the slide.  We use
poly-L-lysine coated slides, subbed manually.  We stain manually for 2
minutes in eosin and then dip 6 times each in 2 changes of 95% ethanol, then
2 minutes in 100% ethanol, etc.  We do hold the racks out of solution to
drain for about 15-20 seconds between steps to minimize carryover.  Could
this be causing the precip?  Is the eosin finding places where drops of
poly-L-lysine didn't drain away and subsequently dried?  Some other reason?

Thanks for your comments.

Regards -
Carol B. McCollough, HT(ASCP)
Diagnostics & Histology Laboratory Manager
Maryland Department of Natural Resources
Fisheries Service
Cooperative Oxford Laboratory
904 S. Morris Street
Oxford, MD 21654

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