Re: clearing problems

From:Melissa Jensen

   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" 
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.
> often find that our first rack into clearing xylene will  not clear ( it
> 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
> 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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