You should neither see a change in scatter, nor in
fluorescence. Just avoid harsh changes in environment e.g.
pre-cool to ~20oC to slow down metabolism and try to sort
out of the medium into medium and use just PBS with no
additives as sheath. Isoton and other commercial sheath
solutions can kill the bugs fast (counterstain with PI at 1
to 10ug/ml).
Make sure that all your media are not only sterile but also
filtered to avoid being drowned in noise, in particular when
rerunning your sorted sample. If possible sort directly
onto agar plates as single cells if you have a cloning
attachment.
Good luck
Gerhard Nebe-v.Caron
Unilever Research, Colworth,
Sharnbrook, Bedfordshire
GB - MK44 1LQ
Tel: +44(0)1234-222066
FAX: +44(0)1234-222344
gerhard.nebe-von-caron@unilever.com
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