Luminex MasterPlex CT User Manual

Tutorial for MasterPlex® CT v1.0

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In this tutorial, we will be highlighting 8 powerful features of the MasterPlex CT v1.0 software:

1) Probe Height Adjustment

2) Wizard Interface

3) Action List Manager

4) Three Step Acquisition

5) Kit Manager

6) Seamless Integration

7) Fail Safe Acquisition

8) Remote Monitor

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Probe Height Adjustment

Being able to adjust the probe height is crucial for any Luminex user as it is the cause for many known issues such as:

 Sample empty errors  Slow event rates  Air in the system  Probes being stuck in the down position  Actuator errors in the software

Adjusting the probe height must also be performed after any of the following events:

 A new type of plate is being used  Probe has been removed (ex. For the declogging procedure)  Luminex system has been moved

You can find the Probe Height launch button in

the left task bar under the Expert Tools menu.

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Thumbwheel

Bolt

The initial Adjust Probe window gives detailed instructions on adjusting the probe height. To adjust the vertical height of the sample probe:

1. Remove the clear plastic shield that covers the sample probe area.

2. Press the Eject button and place a plate in the plate holder.

3. Place the appropriate alignment tool in the plate:  For a standard plate with flat-bottom wells – stack two of the larger

(5.08 mm diameter) alignment discs together and place them into a selected well

 For a filter bottom plate – stack three of the larger (5.08 mm diameter)

alignment discs together and place them into a selected well

 For a half-volume plate with flat-bottom wells – stack two of the

smaller (3.35 mm diameter) alignment discs together and place them into the selected well

 For a round-bottom (U-bottom) plate – stack two of the smaller (3.35

mm diameter) alignment discs and place them into a selected well

 For a plate with conical wells – place one alignment sphere into a

selected well

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4. After you have placed the appropriate alignment tools into the well, press the

Location button and specify the well where you have placed them.

5. Now loosen the bolt that holds the probe (refer to the picture in the Adjust Probe

window) with the hex driver that is provided within the left panel of the Luminex system where the heater block is stored.

6. Click on the Probe Down button to lower the probe arm. The probe height will

get adjusted by the spacers in the well.

7. Using the thumb wheel, lower the sample probe until it rests on, but does not

apply pressure to, the top of the alignment disk or sphere.

8. Tighten the bolt back up with the hex driver.

9. Click the Probe Up button to raise the sample probe.

10. Manually open the XYP door, and click on the Probe Down button again.

11. Watch the black XYP tray to see if it moves. If it moves, repeat this process until

no movement is observed.

12. Click the Probe Up button to raise the sample probe.

13. Replace the plastic shield that covers the sample probe area.

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Wizard Interface

MasterPlex CT provides a set of wizard driven interfaces to help familiarize new users with the software. Whether you are new to MasterPlex CT or new to the Luminex 100 platform in general, wizards will guide you through every step of the way.

There are a total of 8 wizards that will automate everything you will have to do from starting up the Luminex system to shutting it down. You can find the wizard launch buttons under the Home tab in the left task bar under Wizard & Automation.

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Here is the breakdown and a brief summary of the wizards:

 The Startup Wizard walks you through the warm up and startup preparation

sequence.

 The Calibration & Control Wizard walks you through using the calibration and

control beads.

 The Region Activation Wizard guides you through selecting the bead regions

used in the multiplex assay.

 The Acquisition Wizard guides you through preparing the configuration settings

of the Luminex system with clear explanations of the various options that are available.

 The Sample Configuration Wizard walks you through specifying the location of

your samples on the 96-well plate.

 The Report Wizard walks you through generating reports related to the Luminex

system such as Calibration History and System Log Messages.

 The Shut Down Wizard guides you through shutting down the Luminex system.

As an example, we will go through the Startup Wizard. Let’s start by clicking on the Startup Wizard launch button. The initial window provides an outline of the steps to be performed. Go ahead and click on Next to proceed.

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Please note that there are detailed instructions provided at each step. The next window checks to make sure that all the devices are connected properly. If there are any red flags, please check to make sure that all the connections are secured

properly and then click the Connect button to try again. Once all the connections are marked by green checks, click on Next to proceed.

The next screen gives a status of the laser warmup period. While the lasers are warming up, you can accomplish other tasks. Click on Proceed to move on to other tasks.

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In the next step of the Startup Wizard, you can perform the Prime procedure. Priming removes air bubbles from the fluidics sub-system of the Luminex device. You can specify the number of cycles for priming by clicking on the Up or Down arrow under Number of Cycles.

Once you have set the Number of Cycles, simply click on the Prime button and priming will automatically begin by the Luminex system. The status bar shows you the progress of the current action. Once the Prime is done, click Next to proceed.

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The next step takes you through the Alcohol Flush, which removes air bubbles from the cuvette. In this step, you can specify the location of your reagent, in this case 70% isopropanol. To do this, click on the Location button, which in this case is currently labeled Reservoir. The

plate window will pop up allowing you to specify the plate type and well location of your reagent. Specify the Number of Cycles for the Alcohol Flush and click on the Alcohol Flush button to execute the command. Once the Alcohol Flush is done, click on Next proceed.

The last step of the Startup Wizard takes you through the Wash step, which clears out debris and dust particles from the system. You can specify the Location of your reagent, which in this case is distilled water, and the Number of Cycles to repeat. Press the Wash button after you have made your settings and the Wash procedure will begin. When you are done with the wash step, press the Next button to proceed.

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You are now done with the Startup Wizard and your Luminex system is

ready for use. After finishing one wizard, you will have the option to move on to the next wizard in the setup process.

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Action List Manager

The Action List Manager lets you customize and automate routine procedures such as Startup, Maintenance, and Daily Shutdown. You can find the Action List launch button under the Home tab in the left task bar under Wizard & Automation

Add Item button

Remove Item button

As an example, let’s create a Startup action list. Let’s begin by highlighting Warm-up from the bottom-left Item List and then adding it to our Custom Actions list on the lower-right by pressing the Right Arrow button located between the two lists. You can use the Left Arrow button to remove steps from our custom action list. Using the same process, add Prime, Alcohol Flush, and Wash steps to our action list.

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