148 BD High Throughput Sampler User’s Guide
Unexpectedly low event
rate, or no events in
plots (continued)
Cytometer malfunction • Manually run a tube of beads and
verify events appear in plots.
• Check the sample voltage
(BD FACSCalibur)
• Check the Bal seal.
Bubble in sample well(s) Gently tap plate to ensure all wells are
bubble-free, then reacquire sample.
Bubbles in flow cell Ensure BD FACS sheath solution with
surfactant is used during plate-based
acquisition.
Insufficient sample or no
sample in well
Ensure there is sufficient sample in
well.
Bubbles in syringe Prime unit. Choose HTS > Prime.
No filter cap [BD FACSCanto and
BD FACSCanto II] Replace filter cap.
Unexpectedly low
number of events
recorded
Insufficient stopping
criteria
Make sure to set the target number of
events high enough that you do not
run out of events before you reach the
software-based acquisition time
(sample volume/sample rate).
High carryover Mixing volume too great Decrease mixing volume.
Too many mixes Reduce number of mixes.
Sample volume too large Decrease the sample volume.
Wash volume too low Increase the wash volume.
Sample coupler not
completely installed
Slide sample coupler on SIT until you
reach a hard stop.
Damaged injection tubing Replace the sample injection tubing.
See Replacing the Sample Injection
Tubing on page 116.
Acquisition Troubleshooting (continued)
Observation Possible Causes Recommended Solutions
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