Debbie,
how is the humidity in your lab?  Have you recently changed alcohol brands
or have started using recycled reagents?
    Here, in San Antonio, during the humid months we have had to add an
extra xylene on the stain line because the moisture in the air was so bad.
We finally had another HVAC put in to control the temp better.

Joe Nocito, BS, HT (ASCP) QIHC
Histology Manager
Pathology Reference Lab
San Antonio, Texas
----- Original Message -----
From: "Melissa Jensen" 
To: 
Sent: Friday, October 11, 2002 9:31 PM
Subject: Re: clearing problems


> Deb
>    I'm unclear...Are you talking about the first xylene after removing
> slides from the drying oven? Or after the H&E and alcohol into that
xylene?
> ----- Original Message -----
> From: "Deborah Faichney" 
> To: 
> Sent: Thursday, October 10, 2002 3:33 AM
> Subject: clearing problems
>
>
> > I am in need of a bit of advice regarding poor clearing of slides.  Our
> > Haematoxylin and Eosin stain has worked fine for years, but we are now
> > experiencing random clearing problems which follow no particular
pattern.
> We
> > often find that our first rack into clearing xylene will  not clear ( it
> has
> > what looks like drops of water all over slide surface) or it causes the
> > xylene to turn milky/cloudy looking.  The second and subsequent rack/s
of
> > slides can often clear perfectly. Also, in a different scenario, the
first
> > rack can be fine and subsequent racks will not clear.  This can happen
at
> > any point during the week. Some weeks we don't have the problem, others
> do.
> > We change the whole series weekly.  It doesn't seem to matter if we
change
> > the solvents on a daily basis either (tried that to help, but it
doesn't).
> > We can clear the slides if they are taken back from the xylene into
fresh
> > ethanol then into fresh xylene, although this doesn't work every time.
> >
> > Any suggestions greatly appreciated.
> >
> > Debbie Faichney
> > Institute of Aquaculture
> > University of Stirling
> > Scotland
> >
> >
> > Our H&E staining method
> >
> > * Xylene 5 min
> > * 100% Ethanol 2 min
> > * Methylated spirit 1.5 min
> > * Water wash
> > * Mayers Haematoxylin 5 min
> > * water wash
> > * 1 % Acid alcohol (to differentiate if necessary)
> > * water wash
> > * Scotts tap water substitute 1min
> > * water wash
> > * 8 part 1% Eosin(aq) to 1 part Putts eosin 5 min
> > * water wash
> > * methylated spirit 30 sec
> > * 100% Ethanol 2min
> > * 100% Ethanol 1.5 min
> > * Xylene 5 min
> > * Xylene (to coverslip from)
> >
> > --
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