BIO RAD ZE5 User Manual page 353

Error
High fluorescence
No events present during
acquisition
Possible causes
Incorrect optical filter in place
Antibody concentration in sample
too high
Inadequate cell preparation or
washing
Cells have naturally high auto
fluorescence
Poor compensation
Bacterial contamination causing
autofluorescence
Secondary antibody cross-
reacting with cells
Probe too high and not in sample
fluid
Acquisition/Event Issues
Troubleshooting steps
Run the ZE5-EYE to ensure
that installed filters match the
filter configuration in Everest
Software. See Using the
ZE5-EYE to Confirm Filter
Choices on page 135.
Prepare a new sample,
ensuring that antibody
titration is correct.
Prepare a new sample.
Adjust PMT voltages to place
cells on scale. See
Configuring Instrument
Settings on page 257.
Run the compensation
process. See Adjusting
Compensation Automatically
on page 295.
Run the Decontamination
wizard. See Decontaminating
the System on page 325.
Evaluate sample preparation.
Recalibrate the probe, using
custom media settings if
needed. See Media Selector
on page 69.
User Guide | 351