Standardizing cytometer using fluorescent microspheres (target channeling?)

From: Annie Bang (banga@smh.toronto.on.ca)
Date: Fri Aug 23 2002 - 13:44:40 EST


We are using a FASCalibur to measure differences in binding of fluorescent-tagged
antibody to "normal control" and "test" platelets.  We would like to eliminate the
need to use a large number of normal controls per assay to establish a 1% positive
region for the histogram statistic.

Could we set up the cytometer daily, using microspheres to set PMT voltages to give a
consistent channel measurement.  Then run 10 (or so) normal controls to establish the
positive region and analyse all other test samples based on this historical positive
region, no matter what day the test samples are run.

Would this eliminate the need to process a parallel normal control for each test sample
on each day?   Is this even a valid procedure to follow?  Any comments, suggestions,
etc. are appreciated.



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