Hi Abby, 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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