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Technote #
6 Calibration has failed what should I do?
Date Updated Category
17-08-07 Calibration / Control Verification
[STarStation 2.0] [STarStation 2.3]
The Instrument Status Toolbar at the top of the Main Window will indicate the progress of Calibration and report on the success or failure. View the Progress Bar in the Instrument Control Scripts tab to check the progress of the calibration of CAL1 and or CAL2 as appropriate.

The Script Report area on the Scripts tab of the instrument control will indicate the success/failure of calibration.

The event rate during calibration can be observed via the Instrument Status bar. This should be around 250 events/sec

The Calibration History of the System can be reviewed from the LXR Library Log folder which is located at C:\Program Files\Luminex\LXR Library\Log on the System PC for STarStation 2.0. For STarStation 2.3, the Calibration Log file is maintained in the Luminex Library Log Folder which is located in the default path C:/Program Files/Luminex/LXR/DataFiles.

The Calibration History can also be viewed via the STarStation Excel Plugin.

If the System fails calibration follow the procedure below:
1. Verify that the correct Lot numbers and Target Values have been used in the reagent manager. The target values are located on the certificate of Quality data sheets that ship with the calibration reagents.
2. Vortex the calibration microsphere bottles and allow the microspheres to reach room temperature.
3. Verify that the correct wells have been selected in the script and correspond to the locations for the CAL1 and CAL2 microspheres.
4. 5 fresh drops of both CAL1 and CAL2 should be used each time calibration is attempted.
5. Remove the sample probe and sonicate the narrow end for 2-3 minutes. The most common cause for calibration faiulure is a blocked or incorrectly adjusted sample probe.
6. Using a syringe, flush the sample probe with distilled water from the narrow end out through the larger end.
7. Adjust the sample probe height upon replacement of the sample probe in the actuator assembly. See Sample Probe Height Adjustment pdf
8. Complete a maintenance wash to ensure that any air bubbles introduced into the sample probe during cleaning are removed.

A maintenance wash comprises:

<3 Backflush operations followed by,

3 Drain and fills,

2 Alcohol Flushes with 70% ethanol

and 3 Washes with water.

Note: a Maintenance wash may be performed using the supplied maintenance script if you have access to a STARQC or utility microtiter plate.

9. Repeat calibration. Verify that the pressure readings are between 6 and 9 psi and the Events/Second rate reaches at least 250 events/second. If the system sheath pressure does not reach above approx 5psi units (because the sheath bottle cap is loose,a faulty air pump or a pressure leak in the system) a calibration may not begin. The

10. Users must possess Power User or administrative access for the windows operating system if using Windows 2000 or windows XP.

If calibration continues to fail contact Applied Cytometry Customer Support.

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