Calibration of the Luminex is essential for optimal performance and day-to-day reproducibility of results. The Luminex should be calibrated each day after the start up procedure is complete and the optics have been warmed up. Also, the cytometer must be re-calibrated if the instrument temperature changes by more than 3º C during the course of the day.
Before calibrating, make sure that optics warm up is complete.

The calibrators (CAL1 and CAL2) are microspheres with stable fluorescent intensities in the CL1, CL2 & RP1 wavelength ranges.
The calibration process uses these microspheres to adjust voltage settings for optimal and consistent microsphere classification and reporter readings over time and across different instruments. Current calibrated settings are automatically applied to any new session.

During calibration the instrument alters the voltage of the detectors to place the calibration reagents into a mean channel number which corresponds to the target value of the calibrant. The software will attempt this process for approx 180 seconds for both CAL1 and CAL2 reagents before it "times out". If either of the calibrants are not "placed" in the required channels prior to the timeout the calibration for that particular calibrant will fail. In some instances particularly when calibrating with a new batch of reagents for the first time, calibration may fail at the first attempt. Should this occur, attempt to calibrate the failed reagent again. If the calibration continues to fail refer to Technote #6 for troubleshooting.

To modify the reporter detector sensitivity using the HIGH RP1, please refer to Technote #11