Re: Clogging in the sorter unit

From: Neal Benson (nbenson@ufl.edu)
Date: Fri Jun 01 2001 - 08:17:35 EST


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



This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:57:41 EST