Fluidigm CyTOF XT, User Manual

Page: 136 / 148

Appendix A: Troubleshooting
Check the Autosampler Air Filter
136
CyTOF XT User Guide
Observation Possible Cause Recommended Action
Tuning failure See the tuning failures observations in this table.
Dirty or damaged
injector Clean or replace the injector.
Improper installation
of the injector Check that the injector is fully inserted into the heater assembly cap
and the injector cap is properly tightened on the injector.
Leaks in the
Autosampler
Module Visually inspect for leaks and check all connections. Follow
instructional video for fluidic line replacement.
Clogs or insufficient
flow Check nebulizer spray.
Manually clean Autosampler fluidic lines and nebulizer.
Filter problematic sample to remove aggregates and restart
acquisition.
Try to dislodge the clog by running a Heavy wash cycle from the
software upper ribbon.
No, low, or unstable signal
during acquisition Insufficient sample Check that the sample is present in the correct location in the
carousel.
Poor event detection Observed antibody
streaking in rain
plot. Stop acquisition, wash sample, and continue acquisition.
If streaking continues, remove the affected channels from the list of
channels to use for event detection in
Template Editor .
For future experiments, titrate antibodies and other reagents such as
iridium and cisplatin.
High levels of
environmental
contamination Stop acquisition. A medium pre-wash cycle runs automatically.
Check contamination levels before resuming acquisition by opening
Data Display window in the CyTOF Software and running the sample
for approximately 20 sec.
If contamination levels remain high, manually flush the probe line,
sample loop, and nebulizer line.
If streaking is still detected perform weekly cleaning with 10% bleach
solution of the lines.
If level of contamination is very high, the sample should not be run to
prevent damage to the detector.
Cell concentrations
too high Dilute sample to the recommended concentration of no greater than
1 x 10
6 cells/mL.