Sterile Sorting

Joseph Webster J.Webster at centenary.usyd.edu.au
Tue Jun 9 18:46:42 EST 1998


Re sterile sorting problem:
Many years ago I had a similar experience with a FACS 440.
We solved it (eventually) by draining all fluid out of the system,
drying it out by flushing with filtered dry air, and leaving it dry
over a long weekend.
At the same time we autoclaved the sheath container and any detachable
bits that could be autoclaved.

Extreme I know, but it worked back then!

We routinely autoclave the sheath containers periodically, as well as
running 70% ethanol through the system each day.
 
Hope this helps,
	Joseph.

At 12:20 PM 8/06/98 -0500, Joanne Yetz-Aldape wrote:
>Hello, Flowers,
>           I am having some difficulties with sterility of cells
>sorted on my FACStar PLUS.  After running 10% bleach and 70% ethanol
>through the system, one investigator still reports having bacterial
>contamination in the sheath flow-through, plating the sheath in
>gentamicin.  Another investigator reported that her cells were
>overrun with bacteria after 4 days in culture with pen/strep, when
>the sorter had been cleaned in the same way, but  after one other
>cell prep had gone through. The weird thing is that I have been
>sorting this one cell prep every week for quite a 
>while,  and there is no contamination in these cultures after 2 weeks
>with gentamicin.  Could there be a bug  I can't remove from the sorter
>that is resistant to pen/strep, and is there a critical level of
>gentamicin that works in this one situation where the sortied cells
>seem clean?  Is there some detergent that I could use to purge my
>system, or is this something that should just be treated with
>antibiotics?  I do add gentamicin to my sheath tank, also.  Are there
>any special sterility problems associated with the turbo sort option??
> Any advice or comments would be welcome.  Thanks.
>                                                          Joanne
>jyetz-aldape at genetics.com




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