Hi Mike and all operators having similar problems,
I highly recommend using a sterile filter (KLEENPAK) from PALL for the high
pressure sheath containers of the sorters. I never ever had any
contamination problems of cell cultures past a sort. I remember once a very
ugly contamination of the sheath fluid within the tank may be due to
impropper refilling procedure. But the Pall-filter retained everything. Now
I refill the sheath-tank within a bench and protect the fittings of the
sheath tubing and air-pressure tubing with precision wipes (KIMWIPES
Lite)soaked with alcohol and wrapped into aluminium foil during the time of
disconnection from the tank. The Pall-filter is autoclavable but I prefer to
reinstall a new one every 6 months. When replacing the filter or refilling
the tank it is very important when you put pressure on the tank to let the
filter propper refill with sheath and remove any air from the inside of the
filter before putting the fluids system selector knob on 'fill' position. By
this you avoid to get a lot of air in your sheath lines and through the
nozzle.
I not even leave the sheath lines with alcohol overnight, just FACSFlow. I
use FACSFlow as Sheath fluid (no problems with cell cultures up to now).
Flush the sample tubing after sorting daily with alcohol, FACSClean as
bleach, destilled water and FACSRinse. All fluids are sterile filtered prior
to use. Leave FACSRinse in the sample tubing after the last flush and put
the installed nozzle in an Eppendorf tube with FACSRinse to prevent drying
out over night. And at least replace the sample tubing regularly. That's it.
With this procedure the sorter is now running since 2 years without any
problems regarding sterility or contamination.
Hope that helps
Good luck
Volker
Volker Eckstein PhD
Dept. of Internal Medicine V
Medical school of the university
University of Heidelberg
GERMANY
volker_eckstein@med.uni-heidelberg.de
-----Ursprüngliche Nachricht-----
Von: Michael Hughes [mailto:MHughes@picr.man.ac.uk]
Gesendet am: Freitag, 9. August 2002 16:43
An: Cytometry Mailing List
Betreff: yeast contaminations
Hello all.
I have been getting low level yeast contamination in my
sterile sorts during the last week or so. I normally leave the Vantage lines
in 70% alcohol overnight and use sterile PBS which is renewed daily as
sheath fluid. I also use a new sheath filter every day. Can anyone suggest
any ways of cleaning the machine?
Thanks for any help, Mike Hughes
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