RE: FW: H & E Staining problems
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From: | "Colbert, Laurie" <LColbert@phsca.org> |
To: | "'Medilex123@aol.com'" <Medilex123@aol.com>, "Colbert, Laurie" <LColbert@phsca.org>, Histonet@pathology.swmed.edu |
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Content-Type: | text/plain; charset="iso-8859-1" |
Glyn,
What processor had the faulty inlet valves?
Laurie
-----Original Message-----
From: Medilex123@aol.com [mailto:Medilex123@aol.com]
Sent: Tuesday, March 21, 2000 12:07 PM
To: LColbert@phsca.org; Histonet@pathology.swmed.edu
Subject: Re: FW: H & E Staining problems
Dear Laurie,
Annoying isn't it!
We too have had this problem in the past and it seems to boil down to water
(or possibly excess alcohol) in the last xylene/antemedia bottle prior to
wax.
One of our (past!) solvent suppliers used to leave the the empty xylene
containers uncapped, open to the weather outside prior to filling with
xylene, so that small amounts of rain water were included in the xylene. It
was such a small amount of water that the Lab Aids changing the machine
didn't see it and emptied the water into the processor too.
Bitter experience has taught us that as little a 1ml of H2O in 2.25 litres
of
xylene will ruin a machine loaded with biopsies because the tissue is
hydrophillic at all points in the processing schedule and preferentially
absorbs water. When water coated biopsies go into hot wax something awful
happens and the results are as we know - dreadful. In the UK this artefact
is
also known as "Pinks Disease" because of the peculiar eosin staining.
On another occasion the inlet valves on the processor were drawing Formalin
into the last xylene and exactly the same thing happened. We proved the
presence of water in the xylene by drawing up xylene from the base of the
processing bottle using a 10 ml glass pipette. There were small "air
bubbles"
in the xylene which were in fact Formalin, and we also saw damp streaks on
the glass too. We proved the point by mixing the machine xylene and a sample
of known pure xylene (from BDH, who can be relied upon to produce good
reagents) with 1ml of Schiffs reagent. If the machine xylene goes pink and
the BDH xylene doesn't, then you have formalin in your xylene and a fault
with the machine.
Before we found this out we looked at everything, fixation, the time GI bxs
were left "standing" prior to fixation, drying on times post microtomy,
drying on temps, staining, water, everything. By the time we'd finished
looking, the artefact went away because either we'd used a batch of xylene
which came in containers left open in a dry period ( we do get them in the
UK!) or we'd changed the xylenes on the machine and there was no Formalin in
it (yet!). Very frustrating.
May I suggest that you check the xylenes/antemedia thoroughly?
If you let me know whether water contamination is present I would appreciate
it. I'm trying to build up an idea of how common this cause of the artefact
is.
Regards,
Glyn Woodward
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