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Appendix A: Troubleshooting
Check the Autosampler Air Filter
CyTOF XT User Guide
135
Observation
Possible Cause
Recommended Action
Makeup gas flow is
out of range.
Check the makeup gas flow and pressure readbacks in the CyTOF
Software
Instrument Configuration tab > Ion Optics
. Ensure that
readings are within the recommended ranges in the table below.
Reading
Normal Operating Range
Makeup gas flow
0.5
–
0.85 L/min
Makeup gas pressure
0.1
–
3 psi
If the values are outside of the normal operating range, contact
Fluidigm technical support.
•
QC failures
•
RSD(s) >3%
•
Sensitivity
•
CV(s) >15%
•
Insufficient
beads (<2 mL)
•
Inconsistent
sample
introduction due
to bubbles,
leaks, or
clogging in the
Autosampler
fluidic lines
•
Poor nebulizer
spray
Recommended Action
RSD(
s)
>3%
Sensitivity
CV(s) fails
>15%
Check that EQ6 beads are
loaded in the correct location
of the carousel. Add beads or
replace if necessary.
IMPORTANT
Be sure to
vigorously shake the bottle of
EQ6 beads before transferring
the beads to the tube.
✓
✓
Check that the default mixing
and stirring settings have not
been changed to make mixing
or stirring less vigorous. This
can impact the degree of bead
settling within the tube.
✓
✓
✓
Check that the probe line,
sample loop, and nebulizer
lines have been installed
correctly. Prime the syringe
pumps and lines by clicking
the
Prime Lines
button in the
Instrument Configuration tab
> Autosampler
. Clean
Autosampler fluidics lines or
nebulizer if operating pressure
is >10 psi.
✓
✓
✓
Check nebulizer spray. It
should appear as a fine
aerosol that leaves the
nebulizer in an even,
symmetrical V-pattern.
Manually clean the nebulizer
or replace it if necessary.
✓
✓
✓
Clean the cones or replace
them if necessary.
✓
✓
One or more of the
subcalibrations is not being
performed. Run full tuning
protocol.
✓
✓