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Technote #
5 Calibration of CAL1l is successful but CAL2 fails
Date Updated Category
17-08-07 Calibration/Control Verification
[STarStation 2.0] [STarStation 2.3]

If Calibration of the Classification Channel is successful this indicates that the Sample Probe height does not require adjusting and that the Sample Probe is most probably free of blockages.

Confirm that the Reporter Calibration Microspheres are within the expiration date printed on the bottle.

During CAL2 calibration confirm that at least 200 events/second are displayed on the event rate indicator (STarSTation 2.0 & 2.3)

Verify that the calibration vials have been vortexed and that 5 drops of calibraton microsphere are used for each calibration attempt.

Verify that the correct calibration target values were entered into the Reagent Manager

Confirm that the correct wells were selected for the calibration microspheres in the software.

Confirm that the Sheath Pressure is within normal range during the calibration process (6-9 p.s.i.) Ensure that the Gortex seal in the waste container cap is not wet and that the waste container cap is vented.

Remove the Sample Probe and flush using a narrow gauge syringe filled with distilled water.

Adjust the sample probe height using 3 disks, See Sample Probe Vertical Height Adjustment.pdf

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.

Confirm that if you are using Windows 2000 (Luminex 1.7 or IS systems) or Windows XP (IS systems Only) that you have power user or adminintrative access privileges to the operating system, since these are required for successful calibration.

If calibration continues to fail contact Applied Cytometry Customer Support.