Re: FW: H & E Staining problems

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From:Tony Henwood <>
To:"HistoNet (E-mail)" <>, "Colbert, Laurie" <>
Content-Type:text/plain; charset=US-ASCII

Dear Laurie,

Try decreasing the time in the alcohols, xylene and wax. Also check 
the slide drying temperature of the oven. Too hot and Blue cells will 
become apparent.
Regards, Tony

> We have had staining and/or processing problems off and on for years, and we
> are presently going through a very bad period where the problem is not
> correcting itself (which it always has in the past).  Our sections have very
> faded nuclei mainly around the edges and an overall hazy look. Several times
> the pathologists have commented that the slides were too pink. 
> For every theory we come up with, we can always disprove that theory.  We do
> histology for five different hospitals, and the blocks are processed on
> different processors.  Our processing schedules are pretty much the same on
> each processor.  But when we have problems, we have problems with all of the
> hospitals' blocks.
> The main problem is with the small biopsies (skin, GI's, prostate needle
> biopsies, cervical biopsies).  We think it may be an over-processing
> problem, but when we got a demo processor so that we could set up a short
> run for the biopsies, we still had problems.  The tissue looked better, but
> not great.  One day the slides look as bad as they ever had.  Our routine
> processing schedule is nine hours long.  We have one formalin, two PenFix,
> three 100% alcohols, three xylenes, and three paraffins.  We use no heat
> except on the paraffins.
> We have approached the problem as a staining problem, but have pretty much
> ruled this out.  When we sent our slides to other facilities for staining,
> they still looked bad.
> We are at a total loss.  Has anyone ever had water problems that caused this
> problem?  We use tap water.
Tony Henwood
Senior Scientist
Anatomical Pathology
Royal Prince Alfred Hospital

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