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For the last 3 years I have successfully been using a Cicero system connected to
an Epics 753 optical bench to make intra-cellular calcium measurements using
Indo-1. The Cicero software is capable of calculating your required mean
channel vs. time and percent of cells above a control value vs. time. In my
system there has never been a noise problem.
The first thing you should check is to see if you have your triggering set up
correctly. I usually trigger on FALS. Since your FACStar Plus is collecting
data acceptably the Cicero box should mimic your FACStar Plus settings. It is
possible to pick up noise if your trigger level is set to low. Look at your
count rate meter on the Cicero box and see if the count rate approximates what
you are seeing on the oscilloscope and on the FACStar Plus ( note: the Cicero
will see some more cells than the FACStar because of its faster electronics). If
your count rate seems to high turn up the trigger level until the numbers make
more sense. Another possibility is that you may have a problem with your peak
hold circuits in your Cicero box. I suspect that this could cause a ratio noise
problem like the one you described. Call Cytomation to help you trouble-shoot
that one.
It seems like the Cicero system is exactly what you require for your analysis.
I hope you can get it to run properly. Good-Luck!
Ira Schieren
Cell Sorter Facility
Howard Hughes Medical Institute
Columbia University
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