198 BD FACSCalibur Instructions for Use
Fluorescence Compensation
Observation Possible Causes Recommended Solutions
Unable to set
compensation
automatically
Event rate below 400 • Mix tube.
• Set the flow rate to HI.
• Add beads to tube for which
event rate appears low.
75-second time limit
surpassed
Adjust compensation manually
(page 74, step 4).
Old mixed beads Prepare fresh mixed beads.
Rerun the software.
Compensation >50% Incorrect target values Contact BD Biosciences for the
latest target values.
Air bubble in flow cell Prime the fluidics (page 58).
Old mixed beads Prepare fresh mixed beads.
Rerun the software.
Compensation >50%
for FL3–%FL4 and/or
FL4–%FL3
Air bubble in flow cell Prime the fluidics (page 58).
Decreased sheath
pressure from loose cap
or leaking reservoir
Tighten the sheath cap and
check the reservoir for leaks.
Rerun the software.
Old mixed beads Prepare fresh mixed beads.
Rerun the software.
Incorrect target values Contact BD Biosciences for the
latest target values.
Red laser misaligned Contact BD Biosciences.
Compensation results
different than expected
Wrong beads for assay
(ie, PerCP-Cy5.5 beads
for LW assay)
Rerun the assay with the correct
FL3 beads.
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