I am wondering if you are getting air bubbles/pockets in the cassettes
themselves. Are you using mesh cassettes? Are you loading the trays with
cassettes into an empty retort and then starting the delayed program? If so,
air pockets could be forming in the cassette and around the tissue pieces.
Since there is no p/v cycle running yet, those air pockets may be just sitting
there and the exposed tissue pieces may be getting dried out.
I suggest that once the retort is full and in ambient mode, open retort and
move the cassette trays around a bit and see if air bubbles are rising from the
cassettes.
When the trays of cassettes sit on the bench in formalin waiting to go on the
VIP, there may be enough movement when moving the container, etc, that any
air pockets are disrupted and the tissues are not exposed to air pockets.
good luck,
amysue ruppert
Marshfield Labs Histology
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Today's Topics:
1. Re: Processing artifact - delayed start (Bacon, Charles)
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Message: 1
Date: Mon, 5 Feb 2024 13:07:41 +0000
From: "Bacon, Charles" <charles.ba...@baystatehealth.org>
To: Verizon wireless <someperso...@yahoo.com>,
"histonet@lists.utsouthwestern.edu"
<histonet@lists.utsouthwestern.edu>
Subject: Re: [Histonet] Processing artifact - delayed start
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We have had 2 reasons why we saw processing issues like this:
1. I recently found out on our VIP 5 they did not turn the level sensors on
during install. These sensors are known to error so often, Sakura tells
technicians to set the default to off. All the processor can sense is pressure
and time. So it may be that the formalin is not filling all the way. You are
noticing this on delayed runs, but on those runs are you often stacking trays?
If so, isolate the cases in the upper trays and review.
2. We found a clogged line. This happens with the formalin lines if you don?t
do the hot water flush often enough. This can be an even bigger issue if you
recycle and re-buffer your formalin (we do not).
Good luck!
Chuck Bacon, HTL(ASCP)CM
Supervisor Histology
Baystate Medical Center
-----Original Message-----
From: Verizon wireless <someperso...@yahoo.com>
Sent: Friday, February 2, 2024 10:37 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Processing artifact - delayed start
Dear Histonet Members,
We have terrible processing artifact if tissue sits in the formalin-filled
retort (at ambient temperature) for too long (more than 10-12 hours) before a
delayed process starts. The longer the wait, the worse it looks. We have Tissue
Tek VIP 5 processors, and we process luminal gastrointestinal biopsies
exclusively. I've attached some photomicrographs of problem cases on the
Histonet Images website (with the same topic title).
This artifact typically affects a few specimens per day (~2% or less), even
though everything is done on the same processor; it may affect all tissue
portions in a cassette or only some of the tissue in that cassette. Some tissue
portions may only have the artifact on one half with the other half looking
perfectly fine. The techs sometimes note the tissue feeling "crunchy" at the
time of embedding and / or at the time of microtomy. These tissues tend to
suffer greater chatter artifact and have trouble sticking to the slides. The
sections look just as bad on recuts as the originals. Re-processing does not
seem to help at all.
If the cassettes sit in formalin in a container outside of the processor for
days before the processor is loaded (with subsequent immediate start), things
look perfectly fine. When we have staff around to start the processor
immediately upon loading cassettes and empty immediately upon completion, the
tissue looks perfectly fine.
Our current processing program is as follows:
1. 10% Formalin, 30 minutes, ambient temp, p/v on 2. 10% Formalin, 30 minutes,
ambient temp, p/v on 3. 65% Alcohol, 10 minutes, ambient temp, p/v on4. 80%
Alcohol, 10 minutes, ambient temp, p/v on5. 95% Alcohol, 7 minutes, ambient
temp, p/v on6. 95% Alcohol, 7 minutes, ambient temp, p/v on 7. 100% Alcohol, 10
minutes, ambient temp, p/v on 8. 100% Alcohol, 10 minutes, ambient temp, p/v on
9. Xylene, 15 minutes, ambient temp, p/v on 10. Xylene, 15 minutes, ambient
temp, p/v on 11. Paraffin, 10 minutes, 60 degrees C, p/v on12. Paraffin, 10
minutes, 60 degrees C, p/v on13. Paraffin, 15 minutes, 60 degrees C, p/v on14.
Paraffin, 15 minutes, 60 degrees C, p/v on
Obviously there are times we absolutely need to use delayed start. I would
greatly appreciate guidance, and I'll be happy to provide any other details
that might be useful.
Sincerely,
Brian Quigley MDLaboratory Director of a GI Pathology Laboratory
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