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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