Re: Eosin bleeding out

From:Shirley Powell

I have found that if you wash in tap water after eosin, this removes the
majority of the excess stain from slides as well as differentiates the shades
of pink to orange, then finish the dehydration steps.

Michael Becker wrote:

> Hello,
> Long time since I posted.  Please if anyone out there in HistoLand can tell
> me what causes and how can I get rid of Eosin counterstain bleeding out of
> frozen tissue sections after they are coverslipped and are drying.  Once in
> awhile this is a problem.  I have not changed any solutions or technique or
> mounting media.  I use the following protocol:
> 1.  Rapid Fixx-10 to 15 secs
> 2.  Tap water rinse x2
> 3. Gill 3 Hemotoxylin 35-60 seconds depending on age of stain
> 4. Tap water rinse at the sink until water is clear
> 5.  Lithium Carbonate Bluing-5 to 7 dips
> 6. 200 proof Ethanol-10 dips
> 7. 1% Alcoholic Eosin-2 quick dips
> 8. 200 proof Ethanol-4 containers-10 dips each
> 9. 200 proof Ethanol-1 container-1 minute
> 10.Xylene Substitute-Histosolve for 30 secs to a minute or longer.
> Coverslip with Histosolve and a Xylene Substitute mounting media-Histomount
> I get all my reagents except Ethanol from Shandon and have been very happy
> with staining quality.
> Any ideas!  Thanks in advance to all who may respond.
> Hope summer is treating you all well.  Have fun.  Be safe.
> Sue Becker HTL (ASCP)
> c/o Dr. Michael J. Mulvaney
> MOHS Micrographic Surgery
> Albany, NY 12203
> "No one knows noses like MOHS!"

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