RE: H & E Staining problems

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From:"Petrilli, Michael" <>
To:"'Colbert, Laurie'" <>, "HistoNet (E-mail)" <>

Laurie, it seems that quite a few of us have or are having the exact same
problem. I am now completely bald from pulling my hair out when we went
through this. 
Since histology has so many variables it is very hard to eliminate
Here at The Syracuse VA, the changes that we made that seemed to "correct"
the problem are as follows:

1.Frequent tissue processor solution changing
2.Water bath temp no higher than 40 c
3.Minimal usage of freezing spray
4.Quickly pick up sections from bath
5.Temperature of running water in stainer between 20-30 c (we have a Hacker
Linear, so we had a water mixing valve/temp monitor put in)

We have not had a problem in over 18 months.
Let me know if you try these suggestions.Good luck...I know how you feel.

> -----Original Message-----
> From:	Colbert, Laurie []
> Sent:	Monday, March 20, 2000 3:08 PM
> To:	HistoNet (E-mail)
> Subject:	FW: H & E Staining problems
> -----Original Message-----
> From: Colbert, Laurie 
> Sent: Friday, March 17, 2000 2:14 PM
> To: ''
> Subject: RE: H & E Staining problems
> 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.
> Laurie Colbert
> Saint Joseph Medical Center
> Burbank, Ca
> (818) 557-5495 
> -----Original Message-----
> From: []
> Sent: Thursday, March 16, 2000 7:27 PM
> To:
> Subject: H & E Staining problems
> Our laboratory uses an automated Leica stainer for H & E.  The pathologist
> has noticed several slides with weak staining around the periphery of the 
> tissue section.  The stain is very weak just along the edge of the tissue.
> The rest of the tissue stains fine.  I checked the temperature of the
> slide 
> dryer and found no problem.  Any ideas how to solve this problem?

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