Technote #
10 My Luminex IS system fails control verification, what should I do?
Date Updated Category
17-08-2007 Calibration/ Control Verification
[STarStation 2.0] [STarStation 2.3]

There are a variety of issues that can lead to control failure. The most common cause of control or calibration failure is a blocked or incorrectly aligned sample probe.If your instrument fails controls, be sure you have run the calibration beads prior to running controls and verify the following:

1) Verify that the correct Lot numbers and Target Values have been entered into the Reagent Manager and that the correct lots were selected when creating the control verification script.
2) Vortex the calibration vials.
3) Verify that at least five drops of control verification microspheres were used.
4) Verify that xMAP Sheath Fluid or other Luminex approved sheath fluid is being used.
5) Ensure that the Gortex seal in the waste container cap is not wet and that the waste container cap is vented.
6) Verify that the correct wells have been selected from the plate for the CON1 and or CON2 reagents.
7) Remove the sample probe and sonicate the narrow end for 2-3 minutes.
8) Using a syringe, flush the sample probe with distilled water from the narrow end out through the larger end.
9) Readjust the sample probe height upon replacement in the arm.
10) Complete a maintenance wash script (or manually perform 3 Backflush, 3 Drain, 2 Alcohol Flush and 3 Washes with water).
11) Repeat control verification. Verify that the pressure readings are between 6 and 9 psi and the Events/Second rate reaches at least 200.The event rate can be viewed in STarStation versions 2.0 and 2.3 via the system status bar. A low event rate encountered during a CON1 or CON2 operation indicates an empty well, diluted microspheres or a blocked or incorrectly adjusted sample probe.

12) STarStation users MUST be logged into the windows operating system with Power User or Administrator access rights for control verification/calibration operations to be successfully recorded. If STarStation indicates a control failure but the control history (see below) indicates a success this indicates that the user does not possess the necessary write access to the program files folder and that they should have the Power User settings on their windows account reset (consult your network administrator or IT representative for further information).

If the system continues to fail, review the system Control Verification History and contact Technical Support.

Reviewing Control Verification History - STarSTation 2.0

The control history information for the system can be reviewed using the STarStation Excel Plugin via the Import Control History... option on the file menu, or the opening the CON1Log.csv or CON2Log.csv files which are located in the C:/Program Files/Luminex/Luminex Library/Log.

The Control History and CON1Log/CON2Log files record the result of each control verification operation that has been performed on the system.

View the record for the most recent control verification operation (CON1 or CON2) and click the corresponding CON or CON1log tab in the excel or csv file.

Example Control history reports generated by the STarStation Excel Add-In are shown below:

Example CON1 History

Example CON2 History

For Classifier Controls (CON1) confirm that the R1-Count, R2-Count, R3-Count, R4-Count and R5-Count columns have a minimum of 1000 events. There should be a “Yes” in each of the 5 region "R1-Success", "R2-Success", "R3-Success", "R4-Success" and "R5-Success" columns. If there is a "No", this indicates that this region failed and there is likely a clog or your probe is adjusted incorrectly.

For Reporter Controls (CON2) confirm that the R1-Count, R2-Count, R3-Count and R4-Count columns have a minimum of 1000 events. There should be a “Yes” in each of the 4 region "R1-Success", "R2-Success", "R3-Success" and "R4-Success" columns. If there is a "No", this indicates that this region failed and there is likely a clog or your probe is adjusted incorrectly.

The table below defines the parameters A1 through D1 for the CON1 and CON2 operations.



Reviewing Control Verification History - STarSTation version 2.3

To review the System Control Verification history start the LXReports utility via the STart menu->Programs->Luminex->LXR->LX Reports

When the LX Reports Utility Launches Select Control History in the Report Selection, select a date range which includes the most recent control operation and select Both in the Success field. Select the required phase , CON1 or CON2 When the required parameters have been selected press the View button. Examples of Reporter and Classification Reports are shown below.

Example CON1 and CON2 reports are shown below