Awhile back I asked about how to stop alcoholic Eosin counterstain from
bleeding out after coverslipping frozen sections for Mohs surgical
specimens.  Thank you to all who responded with various ideas and
information.  I tried a couple things and voila-no more eosin bleeds.  I
added distilled water after the lithium bluing and I am keeping a better eye
on the xylene substitute clearing agent for signs of water.  I also use a
pre-rinse in a smaller container of xylene sub. clearing before it goes in
the last large container of clearing agent.  This short rinse helps trap any
water so I am not wasting the solvent at the end.  I try to change that one
once every 2 weeks unless the humidity has been really bad.  We are a small
lab but I still try to save the doctor a penny or two here or there if I
can. Reagents and solvents are expensive!  Again thanks to all even to those
whose ideas I did not use.  It is always helpful to hear from more people
than you  really need as you never know whose idea will work.  Alot of you
said the humidity would be a problem and a couple mentioned the pH of the
bluing affects Eosin staining so use a distilled water rinse before the
alcohol rinse and Eosin stain.  These two things are making a big
difference.  Thanks again.  Happy cutting!!

Sue Becker HTL(ASCP)
Histotechnologist
Employed by Dr. Michael J. Mulvaney
Mohs Micrographic Surgery
Albany, NY 12203