 |
 |
 |
 |
i have a question regarding measurement of intracellular calcium using a FACStar
Plus with consort VAX V5.3 and kinpro/cotfit for analysis.
some background:
the starting cell population is cultured dendritic cells with some contaminating
lympocytes and we interested in changes in intracellular calcium when stimulated
with a panel of chemokines. we use indo-1, 30mW of uv (argon ion lines), 405 and
485 filters with pulse processing on and linear scales.
We get a > 4 fold increase in the ratio of violet/blue using ionomycin.
Kinpro is used for kinetics analysis (kinetics smoothing is on and data is
collected every second but only 5 second time points are used in the analysis)
and an analysis gate is set around large cells and also excludes unloaded cells.
Data collection is for six minutes and we collect a total of 100,000 events.
question:
my question is regarding stability (unstability) of the baseline.
the base line is not very stable and i am wondering is that due to the analysis
or the instrument set-up. is it possible that in 10 seconds that i am missing
the peak.
any help would be greatly appreciated
thank you very much
Frank isdell
Flow Cytometry
The Rockefeller University
212-327-7657
|
|
|
|