From: Gerhard Nebe-von-Caron (Gerhard.Nebe-von-Caron@Unilever.com)
Date: Thu Sep 10 1998 - 16:08:26 EST
Dear Colleagues
Please consider the differences in instrument design of the
fluidics. If you for example fill up the sheath filter of an
Elite with bleach it will leak out of there for quite some
time. Your best bet is to disinfect the pipelines somewhere
in the sheath line close to the flow cell and put in a fresh
sterile 'disposable' sheath filter right in the same place
after rinsing with etoh. This is how I sort bacteria on
enrichment and recovery media w/o contamination with a
minimum of expensive operator handling (and otherwise
potentially wasted) time compared to the price of the
appropriate sterile filter. However, consider the potential
reduction of sheath pressure at the flow cell caused by such
a filter on your fluidics / sort-system behaviour.
Regards
Gerhard
______________________________ Reply Separator _________________________________
Subject: Re: sterilizing sorters
Author: trotter@scripps.edu at INTERNET
Date: 08/09/1998 19:37
Tony,
We use only ETOH (70% in dH2O), dH2O, or PBS in the sheath tank/line
system. However, I have used 10% bleach every so often in the sample
line for a minute or so, followed by 2 washes of dH2O, a wash of 70% ETOH,
and then backflushing with sterile (filtered) sheath buffer. We have never
had any sign of toxic effects, but I have only done the bleach wash to
remove what I believed to be biological debris in the line,
not to sterilize the system (and not very often). If something is
really wrong in the sample line we just replace it and use ETOH and
sterile PBS to wash it before putting on cells.
The first thing we did once we saw an apparent toxic effect
to the cells once sorted was to 1) incubate them in different PBS
buffers for a few hours and 2) put them back in the incubator. The cells
grow just fine once exposed to the PBS we made and the BioSure isotonic
saline, but die (slowly) after exposure to the Criterion diluent 2. So,
we're talking about a toxic buffer, not a toxic sheath line. That lot
of Diluent 2 is now only being used in the FACScans.
Regards,
Joe
On Thu, 3 Sep 1998, Antony Bakke wrote:
> Joe,
>
> There has been some discussion on the purdue group lately about sterilizing
> including mention that ethanol does not kill spores, but bleach does. Because
> of that I decided to switch to bleach, but unfortunately found that even hours
> of running sheath did not remove all the bleach, since the sorted cells were
all
> dead. How do you sterilize your instruments and if you use bleach, how do you
> insure that it is removed?
>
> Thank you,
> Tony
>
Joe Trotter
Director, Flow Cytometry
Mailstop Imm-20
The Scripps Research Institute
10666 North Torrey Pines Rd.
La Jolla, California 92037
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