Contact Us

The Low Event Count metric is used to determine the number of events counted during a QC test.

Common Causes for Bead Count Failure:

Bead Preparation and Storage

• We recommend using a bead prep no longer than 5 days and should be stored at 2-8 degrees C.
• Beads should be prepared by adding 2 drops of beads to 300 uL of Buffer Solution (containing 1% PBS and 0.1% BSA) in a 1.5 mL microcenterfuge tube and placed in a vortexer just prior to sampling.

Old probe and tubing

• The probe and tubing can become degraded or clogged after a time, we recommend it be changed after 20 hours of use.

Fluidics Maintenance

• Fluidics Maintenance is a procedure in which the in-line sheath filter, fluidlink and bottle filters are changed out. We recommend this be performed after 176 hours of runtime or every 2 months.

Recommended Troubleshooting:

• Replace the probe and tubing followed by a probe and tubing clean.
• Perform Fluidics Maintenance if it has been close to or over 176 hours of runtime since it was previously performed.
• Run the following cleaning procedures:

– Detector Unclog
– Detector Debubble
– Detector Quick Clean

• Make a new bead prep by adding 2 drops of beads to 300 uL of Buffer Solution (containing 1% PBS and 0.1% BSA) in a 1.5 mL microcenterfuge tube and placed in a vortexer just prior to sampling and run QC.

The Bead Percent metric is used to judge the overall cleanliness of the instrument.  If greater than 25% of the events registered during the QC test fall outside the main gated bead population, it’s an indication of a dirty instrument.

Example of Failed Bead Percent Metric:

Common Causes for Bead Percent Failure:

• Degraded or contaminated probe and tubing
• Contamination of fluids on fluidics station and sheath bottle
• Degraded bottle filters and/or in-line sheath filter

 
Recommended Troubleshooting:

• Replace the probe and tubing followed by a probe and tubing clean
• Replace all fluids or cartridges on the fluidics station
• Empty, decontaminate and re-fill sheath bottle on the bottle station
• Perform Fluidics Maintenance if it has been close to or over 176 hours of runtime since it was previously performed
• Run the following cleaning procedures:

– Detector Unclog
– Detector Debubble
– Detector Quick Clean

• Make a new bead prep by adding 2 drops of beads to 300 uL of Buffer Solution (containing 1% PBS and 0.1% BSA) in a 1.5 mL microcenterfuge tube and placed in a vortexer just prior to sampling and run QC.

The CV metric judges the width of the laser as it hits the flow cell.

Example of Failed CV Metric:

Common Causes for CV Failures:

• Instrument needs routine maintenance
• Leaking at the probe and tubing-fluidlink connection or fluidlink-detector connection.
• Instrument vibration caused lasers to widen

Recommended Troubleshooting:

• Replace the probe and tubing followed by a probe and tubing clean.
• Perform Fluidics Maintenance if it has been close to or over 176 hours of runtime since it was previously performed.
• Run the following cleaning procedures:

-Detector Unclog
-Detector Debubble
-Detector Long Clean

Run a 4-minute Prime and check for leaks at the probe and tubing-fluidlink connection or fluidlink-detector connection.

• If leaks are found at the probe and tubing-fluidlink connection, please reach out to Support@Sartorius.com
• If leaks are found at the fluidlink-detector connection, please ensure the fluidlink is threaded correctly and securely tightened.
• Make a new bead prep by adding 2 drops of beads to 300 uL of Buffer Solution (containing 1% PBS and 0.1% BSA) in a 1.5 mL microcenterfuge tube and placed in a vortexer just prior to sampling and run QC.

The Top Peak Mean (TPM) metric judges the voltage set by the PMT modules in the instrument.

 
Common Causes for TPM Failures

• Instrument needs routine maintenance
• Leaking at the probe and tubing-fluidlink connection or fluidlink-detector connection.
• Instrument vibration caused lasers to widen.

 
Recommended Troubleshooting:

• Replace the probe and tubing followed by a probe and tubing clean
• Perform Fluidics Maintenance if it has been close to or over 176 hours of runtime since it was previously performed
• Run the following cleaning procedures:

– Detector Unclog
– Detector Debubble
– Detector Long Clean

Run a 4-minute Prime and check for leaks at the probe and tubing-fluidlink connection or fluidlink-detector connection.

• If leaks are found at the probe and tubing-fluidlink connection, please reach out to Support@Sartorius.com
• If leaks are found at the fluidlink-detector connection, please ensure the fluidlink is threaded correctly and securely tightened.
• Make a new bead prep by adding 2 drops of beads to 300 uL of Buffer Solution (containing 1% PBS and 0.1% BSA) in a 1.5 mL microcenterfuge tube and placed in a vortexer just prior to sampling and run QC.

Instrument clogging can cause a QC test to fail multiple metrics.

 
Common Causes for Clogging:

Clogging can be the byproduct of multiple factors such as poor instrument cleanliness, old or clogged consumables, degraded filters, sample type, sample preparation, or experimental design
 
Example of Failing QC Due to Clog:

• The bead population will present itself in a smear pattern going from bottom left to top right.

 
Troubleshooting a Clog:

• Change out the probe and tubing and run a probe and tubing clean.
• Perform Fluidics Maintenance if it has been close to or over 176 hours of runtime since it was previously performed.
• Run a Detector Unclog cleaning procedure.
• Make a new bead prep by adding 2 drops of beads to 300 uL of Buffer Solution (containing 1% PBS and 0.1% BSA) in a 1.5 mL microcenterfuge tube and placed in a vortexer just prior to sampling and run QC.

Air can be introduced into the system and cause a QC test to fail multiple metrics.

 
Common Causes for Air in the System

Air can be introduced into the system through loose connections on the bottle station or in-line sheath filter.
 
Example of Failing QC Due to Air:

Troubleshooting Air in the System:

• Check that all connections on the bottle station are secure.
• Confirm all tubing lines are going to the corresponding bottles on the bottle station.
• Check connections on in-line sheath filter are secure.
• Perform a Detector Debubble cleaning procedure.
• Make a new bead prep by adding 2 drops of beads to 300 uL of Buffer Solution (containing 1% PBS and 0.1% BSA) in a 1.5 mL microcenterfuge tube and placed in a vortexer just prior to sampling and run QC.