You can always look up the archive of the purdue mailserver.
As long as the search engine of the pudue site is still
under construction you can still use alta vista (search:
sorting + sterility + purdue) and will actually find a lot
of the purdue messages from peoples archives and the server
itself.
One of the key points to sterility is to insert a disposable
fresh sterile filter into the sheath line closed to the flow
cell. On the Elite I suck 50ml of 10% Domestos through the
sheath line behind the filter (apply vacuum to the flow
cell)followed by 5ml 75% isopropanol. I then replace the two
sterile filter units (Millifil GS #SVGSL10RH from Millipore)
I connected in parallel into the sheath line to reduce
pressure loss / increase filter surface. The sterile filters
have to be replaced before every critical sort. As I can
sterile sort bacteria onto nutrient agar I have no doubts
about the efficiency of the process. The whole thing takes
less than 20 minutes, the time the machine has to warm up
anyhow. It avoids residual disinfectant in the pipework and
missing deadspaces in the -these days rather complex-
pneumatic and fluidic system.
Last step is to run a 10% domestos sample to treat the
sample line.
To reduce airborn contaminations we actually had a cardboard
cover above the sort area to reduce things falling from the
ceiling (or air-condition) into the medium.
Good luck
Gerhard
______________________________ Reply Separator
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Subject: Sort Sterility
Author: opt074@abdn.ac.uk at INTERNET
Date: 11/03/1999 20:14
A few months ago there was quite a lot of disscussion on
the best reagents etc. to use to reach optimal cell
survival and sterility after cell sorting. If possible,
could someone remind me of the conclusions the discussion
came to??
Cathryn
----------------------
Cathryn Broderick
c.broderick@abdn.ac.uk
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