In the Luminex Assay Tutorial Level II
, we import a batch of runs
but gloss over the Define Well Roles
section of import. The checkboxes there are used to identify titrations and single point controls you want to be able to use later. Return to this view by beginning to import (or reimport) any run. After entering run properties, you will see:
Standard vs. Quality Control
In the Define Well Roles
section, you can mark a titration as providing both a standard and a quality control. If you check both, you will see twice as many curves (see Step 4: View 4pl and 5pl Curve Fits
). Standard titrations are used to calculate the concentrations of samples and the values displayed in Levey-Jennings plots (see Step 5: Track Analyte Quality Over Time
). In contrast, quality controls are used in a lab-specific manner to track the performance of the assay over time. For this tutorial, we designate our titrated standards as standards, but not as quality controls.
In order to have values for EC50, AUC, and other calculations done without adding the selected titration to the Levey-Jennings plot and QC Report, select the Other Control
checkbox. This is useful for titrations that will not be tracked using these reporting tools, but still need the transform script to calculate values.
Multiple Standards per Analyte
If the data for your run included multiple standard titrations, you would be able to choose which analytes to associate with which standards. Note that you must have marked each standard as a standard in the Define Well Roles
section before it will appear as an option. Each standard will appear over a column of checkboxes in the Analyte Properties
section. You can select more than one standard for each analyte by selecting the checkboxes in more than one standard’s column. The data for this tutorial includes only one standard, so this option is not explored here.
To track performance of a single-point control over time, such as in the case of an antigen panel where the control is not titrated, you can select Tracked Single Point Controls
. Check the appropriate boxes in the Define Well Roles
panel during run upload. This feature is explored in Track Single-Point Controls in Levey-Jennings Plots
During import of Luminex data, if a sample appears at different dilutions, we infer that it has been titrated and the user can indicate whether they are using the titrated sample as a standard, quality control, or unknown.
When multiple plates of data are uploaded together in a single run
, LabKey will infer cross-plate titrations
when either of the following conditions are met:
- If the value in the Description field is repeated 5 or more times in the "Summary" section, or
- If the Description is repeated 10 or more times in the "Raw" section.
These identified potential cross-plate titrations are assigned a name matching the description field that identified them, and the user confirms during upload if they are indeed titrations using a checkbox. Uncheck any incorrect inferrals.
The titration data is then used for result calculations and further analysis and plots.