Flushing the lines with water is a good practice to avoid the problem; trouble is, when your lab is busy, it often is skipped or not done thoroughly enough. There's a continuous fluid connection at least to the valve, and salt from your sheath buffer will diffuse into the water over time. (Perhaps some bio-waste gets in there too.) We used to have the same problem when the sort unit was not used regularly. Our solution: at least once a week, put a tube of buffer on the sample port, set a sort gate, and do an "empty" sort of at least 10 ml into each collection tube. It'll take you around 5 minutes or less. That keeps the fluid moving along and easily removes the salt buildup before it gets too bad. We have student-assistant who performs routine lab maintenance, and that's just one of her regularly scheduled jobs. Neal Whist Solvejg Knagenhjelm wrote: > > Dear colleagues! > > I am planning to do some cell-sorting (ovine granulocytes) in connection > with the trial that I am working on at the moment. I use FACSCalibur for > flow cytometry and cell sorting. This machine has been used for sorting > previously though the last time was in October. Since then noone has used > the sorting unit at all. BD has informed us that we should rinse the sorting > unit/tubes after use with distilled water; which has therefore been done > every time. Now that I am going to try out the sorting unit mayself I find > that the tube going from inside the flow cytometry leading to the unit is > completely clogged with salt-crystals. I deduce from this that the unit has > not been thouroughly rinsed the last time it was used. Since nothing seemed > to be able to dissolve or move the clog, we called BD to have this tube > replaced. Their people inform us that sheath-fluid goes into these tubes > every time we use the flow cytometer..even if we are not using the sorting > unit. In none of the manuals or the information we have recieved from their > personell does it say that this happens, and there are no procedures > described for it..this the BD technicians admit, they have just seen that it > happens all the time. They say that one has to rinse these tubes at least > every time one does a system-clean (monthly) and also keep an eye on these > tubes all the time to see if there are any crystals forming. > We don't really believe them, as we didn't start using the sorter before we > had had the machine over half a year and no crystals had formed in the tubes > in this period. The sorter has not been used regularly either and no > crystals have formed between use (expect for this last time) either. > Have any of you experienced this problem and have gotten this kind of > information from BD (that one has to rinse the tubes often even when not in > use)?? > Appreciate any thoughts/comments! > Regards, > Solvejg K.Whist, DVM, PhD-student > The Norwegian School of Veterinary Science > POBox 8146 Dep., > N-0033 Oslo > Norway > Fax: +47 22964818 > e-mail: solvejg.k.whist@veths.no > > ______________________________________________________________________________________________________________ > > This message has been scanned for viruses and harmful content, and found > secure. > > Norwegian School of Veterinary Science > > ______________________________________________________________________________________________________________ -- Neal Benson University of Florida nbenson@ufl.edu
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