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The probable cause was the protocol settings left over from a previous
run. The FACSCan/LYSYS combination is very versatile, and even a
methodical person like me can't always remember all the details every
time.
Some rules & procedures I set up in our multi-user facility have been
successful in getting the data saved as expected:
1) I recommend a routine calibration every day! We use BD CaliBrite, but
others will do fine; The main point is that you have a known reference
from which to compare.
At very least, there should be a standard instrument settings file, but
that will not help if/when your instrument performance changes.
2) ALWAYS quit and restart LYSYS between runs! ALWAYS!
That way there are no surprises left over from the last run, (like a
protocol with FL2 & FL3 turned off!)
3) Keep your own protocol, Reagent List, Instrument Settings etc. (if
necessary, a set for each type of experiment) saved on a floppy &/or in
your own directory. Then you can simply recall them and not have to
remember to reset EVERYTHING!
In the scenario I suggest, the FL2 data will appear on the screen while
acquiring, but will not be saved if the protocol says "off" for that
parameter.
Good luck, and May the Flow be with You!
Joseph Webster
Centenary Institute
Locked Bag No 6
Newtown NSW 2042
AUSTRALIA
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