> > ----------
> > Van: Jeff_Carrell@hgsi.com[SMTP:Jeff_Carrell@hgsi.com]
> > Antwoord naar: Jeff_Carrell@hgsi.com
> > Verzonden: woensdag 6 oktober 1999 21:41
> > Aan: Cytometry Mailing List
> > Onderwerp: sheath tank filling
> >
> >
> > Hello All,
> >
> > Does anyone fill their cell sorter sheath fluid tank "out in the
open", or
> > does
> > everyone fill the tank in the laminar flow cabinet?
> >
> > Just curious,
> >
> > Jeff Carrell
> > Human Genome Sciences
> >
Greetings Jeff.
I have been sorting for several years on Coulter machines (EPICS C, ELITE)
during that time I estimate that I have performed several hundred sterile
sorts. The contamination rate over this time has been precisely three
episodes (and I have a strong suspicion that these were due to
manipulations performed on the sample post sorting). Most of these samples
were put back into culture with no antibiotic addition.
Here is my updated regime for sterile sorting:
Routinely, I use purified (Reverse Osmosis)water as the sheath medium when
I am not sorting.
The Coulter instruments have a 'rinse' reservoir which I fill with 0.2%
benzalkonium Chloride ('Roccal')/ (+1% Sodium Nitrite as a corrosion
inhibitor) in purified water.
Every six months or so I replace the in-line sheath filter, when I do this
I exchange the filter with a 'straight through' connector and flush the
system with a surface-active agent (2% 'decon 90') followed by several
changes of water.
Prior to sorting, I take the sheath reservoir apart on the bench (no
special precautions), fill it with 3litres of R.O water straight from the
machine and add 30 X10 PBS tablets (OXOID Bri4a). When they are dissolved,
I put the tank back together, put it into the machine, turn the key and go.
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Arnold Richard Pizzey
Department of Haematology
Royal Free and University College London Medical School
98 Chenies Mews
London WC1E 6HX
U.K
voice: +44 171-209-6234
Fax: +44 171-209-6222
email: a.pizzey@ucl.ac.uk
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