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well we have two FACScans running in a >100 user environment, and when I
arrived in Pasteur we had the same problem as you. I've solved it pretty
much in the following way:
1. I require from everybody to come with clean sample preparations (filter
them through 50u nylon mesh before the stainings). If I see people running
with more than 1000 events/sec, or with sample preparations that contain
clumps of RBC's or dead cells, they are in risk of loosing access...
2. I require everybody to clean up the machine with 1 min FACSRinse (or 1%
dishwasher detergent works as well), followed by 1 min PBS, and if they are
the last that day, followed by 2 min dH2O.
They have also to let 2 ml of each to aspirate through the bio-hazard pump
(with the lever to the right...) before running it through the flowcell.
This way also the bio-hazard tubing is cleaned up (they tend to clog from
time to time causing terrible backflush contamination of the samples).
3. We clean the machine in the middle of the day (from 3:30-4 pm) for 1/2 h
with FACSRinse (or detergent) and PBS + dH2P.
4. I rarely have to do a monthly cleaning.
This way I have rather rarely clogging of the machine, and if, it's mainly
because some new users don't prepare their cells right (so it's enough to
talk to them seriously, and they'll stop it in most of the cases).
Here are the reasons why I do it like this:
1. Clean sample preparations are the MOST important factor in keeping the
machine clean. Everything else is rather secondary (washing etc), compared
to this point (and that's the one most people ignore as well...).
2. I opted for short rinse periods, rather than longers, as long cleaning
tend to discourage people to do it at all, or they leave the machine
running unattended, and run it dry (and that's worse than no cleaning at
all).
3. Cleaning in the middle of a 10h acquisition day, helps us to prevent
problems rather than to wait until its too late. Have to admit that this
might be difficult to have somebody do, if you have nobody responsible for
the machines.
4. Because I'm convinced that my cleaning protocol is largely sufficient...
Of course if people run bioharzardous samples (Acridine Orange, human
samples etc) they are required to clean and decontaminate appropriately...
Hope that might help you to get your machines running ok..
Cheers, Matthias
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web