Fluidigm CyTOF XT - XY Alignment Failures; Detector Voltage Optimization; Current Optimization; Nebulizer Gas Flow Issues

Appendix A: Troubleshooting
Check the Autosampler Air Filter
134 CyTOF XT User Guide
Observation
Possible Cause
Recommended Action
Check the pressure on the push pump during injection. If it is >10 psi,
clean the nebulizer. If pressure remains high, clean the Autosampler
fluidic lines.
If signals remain low, manually tune XY alignment and current. Then
restart autotuning.
Rerun tuning. If mass resolution fails again, contact Fluidigm
technical support.
• XY alignment
XY Calibration failed
(no optimum found)
1 Rerun tuning, applying the settings from the most recent passed
tuning. Refer to the CyTOF Software v8.0 Help for CyTOF XT
(FLDM-00045)
2 Manually tune XY alignment. Refer to the CyTOF Software v8.0
Help for CyTOF XT (FLDM-00045)
• Detector voltage
optimization
No optimal voltage
found
Rerun tuning.
Optimal voltage is
out of range.
Monitor detector voltage weekly and when it reaches 1,000 V
contact Fluidigm technical support to schedule a replacement of the
detector.
• Current
Current optimized
to 10
Clean cones. If current value is still 10 after cleaning, replace cone(s).
• No nebulizer gas
optimum found
Nebulizer gas flow
out of specification
Check the nebulizer spray with a flashlight. It should appear as a fine
aerosol that leaves the nebulizer in an even, symmetrical V-pattern. If
it is not, clean the nebulizer or replace it if necessary.
If the nebulizer pressure is higher than the range specified, the
nebulizer may be blocked. Clean the nebulizer or replace if needed.
Check the nebulizer gas flow and pressure readbacks in the CyTOF
Software Instrument Configuration tab > Ion Optics. Ensure that
readings are within the ranges recommended in the table below.
Reading
Normal Operating Range
Nebulizer gas flow
0.150.3 L/min
Nebulizer gas pressure
1218 psi
If the nebulizer pressure is outside of the normal operating range,
manually adjust the nebulizer gas flow (within the range specified
above) until you obtain a reading between 12 and 18 psi (optimal 15
psi).
If even after manual adjustment of the nebulizer gas, the pressure is
too low, either there is a gas leak, or the nebulizer has aged and
needs replacement. Check that the nebulizer gas line is connected
properly. Check the Nebulizer Holder O-ring for damage and replace
if necessary. If the optimal nebulizer gas flow is >0.3 L/min, replace
the nebulizer.
• No optimal makeup
gas found
Oxides failure
during tuning
Possible leak in the spray chamber or nebulizer gas line. Check
nebulizer gas line. Check spray chamber O-rings for damage and
replace them if needed.