Table of Contents

guest
2022-05-20
       Luminex Assay Tutorial Level II
         Step 1: Import Lists and Assay Archives
         Step 2: Configure R, Packages and Script
         Step 3: Import Luminex Runs
         Step 4: View 4pl and 5pl Curve Fits
         Step 5: Track Analyte Quality Over Time
         Step 6: Use Guide Sets for QC
         Step 7: Compare Standard Curves Across Runs

Luminex Assay Tutorial Level II


LabKey Server's tool for Luminex® assays can help you to manage, quality control, analyze, share, integrate and export Luminex results.

This tutorial builds on an understanding of material covered in Luminex Assay Tutorial Level I and shows you how to:

  • Import Luminex assay data stored in structured Excel files that have been output from a BioPlex instrument.
  • Set values during import of pre-defined analyte, run and batch properties for each analyte, run and/or batch of runs.
  • Run the LabKey Luminex transform script during import to calculate logistic curve fits and other parameters for standard titrations.
  • View curve fits and calculated values for each standard titration.
  • Visualize changes in the performance of standards over time using Levey-Jennings plots.
  • Determine expected ranges for performance of standards for analyte lots, then flag exceptional values.

Tutorial Steps

Note that for simplicity, the sample datasets used in this tutorial include only standard titrations. All steps covered here could equally apply to quality control titrations.

Further detail and background on many of the steps in this tutorial can be found in the Luminex Reference documentation

The Luminex assay data you will import in this tutorial can also be seen in the interactive example. There you can explore features that do not require editor-level or higher permissions.

First Step




Step 1: Import Lists and Assay Archives


In this step, we import two pre-prepared archives to simplify creating a more complex assay design than we used in the first tutorial. These define:
  • A set of lists used by the assay design to define lookups for several properties
  • A pre-prepared assay design for a Luminex┬« experiment

Set Up

  • If you have not already set up the Luminex tutorial project, follow this topic to do so: Set Up Luminex Tutorial Folder.
  • Return to this page when you have completed the set up.

Import the List Archive

The lists imported here define sets of acceptable values for various properties included in the assay design that you will import in a later step. These acceptable values are used to provide drop-down lists to users importing runs to simplify data entry.

Import the List Archive

  • Navigate to your Luminex tutorial folder.
  • Select (Admin) > Manage Lists.
  • Click Import List Archive.
  • Click Browse/Choose File and from the Luminex sample data you downloaded, select /Luminex/Luminex_ListArchive.lists.zip.
  • Click Import List Archive.

Import the Assay Design Archive

Next, you will import a pre-prepared assay design which will be used to capture Luminex data.

  • Click the Luminex link to return to the main folder page.
  • In the Files web part, double click the Luminex folder.
  • Select Luminex Assay 200.xar.
  • Click Import Data.
  • In the popup dialog, confirm that Import Experiment is selected and click Import.
  • Refresh your browser, and in the Assay List, the new assay design "Luminex Assay 200" will appear.

Start Over | Next Step (2 of 7)




Step 2: Configure R, Packages and Script


Luminex® analysis in LabKey Server makes use of a transform script to do a number of calculations including curve fits and estimated concentrations. To get the transform script running, you will need to:

Install and Configure R

You will need to install R and configure it as a scripting language on your Labkey Server. If you are on a Windows machine, install R in a directory that does not contain a space (i.e. not the default "C:\Program Files\R" location).

Install Necessary R Packages

The instructions in this section describe package installation using the R graphical user interface.

Install Ruminex

The transform script requires Ruminex, a custom package not available on CRAN, so you will need to use the zip package provided in the sample files you downloaded. This tutorial was tested using Ruminex 0.1.0 (and 0.0.9); other versions may not work identically.

  • If you are running R-3.0.0 or later, use Ruminex_0.1.0.zip from the LuminexSample files you downloaded. (Or download it here: Ruminex_0.1.0.zip).
  • Launch the R graphical user interface. Use the drop-down menus in the R user interface to select: Packages > Install package(s) from local file...
  • Open the zip file you downloaded from this page.

Install Additional Packages

  • Open your R console.
  • Using the R console, install the packages listed below using commands like the following (you may want to vary the value of repos depending on your geographic location):
install.packages("Rlabkey", repos="http://cran.rstudio.com")
  • Install the following packages (some may be downloaded for you as dependencies of others):
    • Rlabkey
    • RCurl
    • rjson
    • xtable
    • drc
    • Cairo
  • As an alternative to the R console, you can use the R graphical user interface:
    • Use the drop-down menus to select Packages > Install package(s)...
    • Select your CRAN mirror.
    • Select the packages listed above. (You may be able to multi-select by using the Ctrl key.)
    • Click OK and confirm that all packages were successfully unpacked and checked.

Associate the Transform Script with the Assay Design

Control Access to Scripts

Next, place the transform script and utility script in a server-accessible, protected location.

Your script was uploaded into the Files web part earlier; however, we do not recommend running it from this location. If you do so, all users with edit-level permissions on your server will be able to replace/edit the script which runs with very high levels of permission on your server. Before continuing, place a copy of the transform script and its associated utility script in a safer location on your server. If your server runs on Windows, for example:

  • Locate the LABKEY_HOME directory on your local machine. For example, it might be C:\labkey\
  • Create a new directory named scripts here.
  • Place a copy of each of these files in this new directory: (Copy from the sample data you already downloaded.)
    • labkey_luminex_transform.R
    • youtil.R

For more information about locating R scripts, see Transformation Scripts in R.

Add Path to Assay Design

Edit the transform script path in the assay design we provided to point to this location.

  • In the Assay List of your Luminex folder, click Luminex Assay 200.
  • Click Manage Assay Design > Edit assay design.
  • In the Assay Properties section, for the Transform Script, enter the full path to the scripts you just placed.
  • Click Save

When you save, the server will verify the script location.

Test Package Configuration

This step is optional, but will confirm that your R script will run as needed in the following tutorial steps.

  • Click Luminex.
  • In the Files web part, select /Luminex/Runs - Assay 200/02-14A22-IgA-Biotin.xls.
  • Click Import Data.
  • Select Use Luminex Assay 200 in the popup and click Import again.
  • For Batch properties, leave defaults unchanged and click Next.
  • For Run Properties, leave defaults unchanged and click Next.
  • Click Save and Finish.

If there is a problem with the path, or with installed packages or the version of R, error messages will help you figure out what else you need to do (e.g., installing an additional R package or upgrading your version of R). After installing a missing package, you can refresh your browser window to see if additional errors are generated.

If your script cannot find youtil.R, make sure it is located in the same directory as the LabKey Luminex transform script. The following should be peers:

  • labkey_luminex_transform.R
  • youtil.R
For further troubleshooting tips, see: Troubleshoot Luminex Transform Scripts and Curve Fit Results

When you see the Luminex Assay 200 Runs grid, you know your script was successfully run while importing the test run.

Delete the Imported Run Data

Before continuing with the tutorial, you need to delete the run you used to test your R configuration.

  • Select (Admin) > Manage Assays.
  • Select Luminex Assay 200.
  • Click the top checkbox on the left of the list of runs. This selects all rows.
  • Click (Delete).
  • Click Confirm Delete.

Previous Step | Next Step (3 of 7)




Step 3: Import Luminex Runs


Here we import 10 Luminex runs to have enough data to provide interesting demonstrations of the tools. You could import each individually as we did in the Luminex Level I tutorial, but using a batch import streamlines the process somewhat.

Import Batch of Runs

  • Return to the Luminex folder.
  • In the Files web part, open the Luminex/Runs - Assay 200 folder.
  • Check the boxes for all 10 run files (the run files begin with a two digit number and end with an .xls extension. Ignore the artifacts with other extensions created when you did a test import.)
  • Click Import Data.
  • Select the Luminex Assay 200 assay design.
  • Click Import.
  • For Batch Properties, leave default values as provided.
  • Click Next.
  • In Run Properties, enter the run number (the first two digits of the file name which is listed in the Run Data field) as the Notebook No.
  • Click Next.
  • In Define Well Roles, leave default boxes checked. For information about these options, see Review Well Roles.
  • In Analyte Properties, edit the Lot Number:
    • Check the Lot Number/Same box to give the same lot number to all analytes.
    • Enter "Lot 437" for all analytes when importing runs #01-05.
    • Enter "Lot 815" for all analytes when importing runs #06-10.
    • Because the previously entered value will be retained for each import, you only need to edit this field when importing the first and 6th runs.
  • In Analyte Properties, check the box in the Negative Control column for Blank.
  • Then check the Subtract Negative Bead/Same box, and select Blank as the value for all other analytes. These properties are explained here.
  • Click Save And Import Another Run. Wait for the page to refresh.
  • You will return to the Run Properties page showing an "Upload successful" message and with the next file name in the Run Data field. Note that the previously entered Notebook No value is retained; remember to edit it before clicking Next, then Save and Import Another Run.
  • Continue this loop entering the new Notebook No for each run and changing the Analyte Lot message at the 6th run to "Lot 815".
    • Note: a log file is written for each upload. If you upload two files in the same minute, you will be warned that there is already a log file with that name. Simply wait and click Save and Import Another Run again until the upload completes.
  • Click Save And Finish when finishing the 10th run.

View the Imported Runs and Data

When you are finished importing the batch of 10 runs, you'll see the fully populated runs list. It will look something like this.

Viewing Data

To see the results for a single run, click one of the links in the Assay ID column on the runs page. For example, if you click 10-13A12-IgA-Biotin.xls, you will see the data just for the tenth (latest) run, as shown here.

Above the grid are links for View Batches, View Runs, and View Results.

  • View Batches: The ten runs we just imported were all in one batch; you might have uploaded other runs or batches as well.
  • View Runs: Shows the list of all runs you have imported in all batches. This is the view you saw when import was complete.
  • View Results: Shows the assay data, or results, for all imported runs in one grid. The results table for all runs has nearly 5,000 rows, as shown here.

Previous Step | Next Step (4 of 7)




Step 4: View 4pl and 5pl Curve Fits


This step introduces some of the values calculated by the server and transform script for each standard's titration, including the 4- and 5-parameter logistic curve fits. For each run, the script outputs a PDF that includes plots for curve fits for each analtye. Each plot shows the dose response curve for fluorescence intensity with increasing concentration or reduced dilution. These plots can be useful for examining how well the curves fit the data.

For additional background and details about these and other calculations that are performed, see Luminex Calculations.

View Curve Fits

As an example, here we view one of the 5pl curves generated for the tenth run.

  • In the Luminex folder, select (Admin) > Manage Assays.
  • Click Luminex Assay 200.
  • Click the curve icon in the Curves column for the tenth run (Assay ID 10-13A12-IgA-Biotin.xls).
  • Select 10-13A12-IgA-Biotin.Standard1_5PL.pdf.
  • Open the file. Depending on your browser settings, it may open directly or download for you to click to open.
  • You will see a series of curves like the one below:
  • You can also see the example by downloading this PDF containing the full set of curves.

Note: The PDF files for these curves for each run were deposited by the LabKey Luminex transform script in the Runs - Assay 200 folder when the script ran during run import.

View Calculated Values

Some calculated values are stored in the results grid with other Luminex data, others are part of the titration qc reports for standards and other titrations. For more information about the calculations, see Luminex Calculations.

View Calculated Values in Titration QC Reports

For the same tenth run, view calculated values including estimated concentration at 50%, or EC50.

  • Return to the Luminex Assay 200 Runs grid. (If it is not still shown in your browser, select (Admin) > Manage Assays and click Luminex Assay 200.
  • In the runs list, click the Assay ID "10-13A12-IgA-Biotin.xls."
  • Click View QC Report > view titration qc report.
  • The report shows one row for each analyte in this run. You can see a similar one in the interactive example.
  • Scroll to the right to see columns calculated by the script:
    • Four Parameter Curve Fit EC50
    • Five Parameter Curve Fit EC50
    • High MFI
    • Trapezoidal Curve Fit AUC

Since you selected the report for a single run, you will see 6 rows for just that run. To see these values for all runs, scroll back to the left and clear the run filter by clicking the .

View Calculated Values in Results Grid

  • Click View Runs.
  • In the runs list, click 10-13A12-IgA-Biotin.xls
  • In this Results view, scroll to the right to see columns calculated by the script:
    • FI-Bkgd-Neg
    • Standard for Rumi Calc
    • Est Log Conc - Rumi 5 PL
    • Est Conc - Rumi 5 PL
    • SE - Rumi 5 PL
    • Est Log Conc - Rumi 4 PL
    • Est Conc - Rumi 4 PL
    • SE - Rumi 4 PL
    • Slope Param 4 PL
    • Lower Param 4 PL
    • Upper Param 4 PL
    • Inflection Param 4 PL
    • Slope Param 5 PL
    • Lower Param 5 PL
    • Upper Param 5 PL
    • Inflection Param 5 PL
    • Asymmetry Param 5 PL

You could also view these values in the interactive example.

Previous Step | Next Step (5 of 7)




Step 5: Track Analyte Quality Over Time


In this step, we will visualize a trend in the performance of a standard using a Levey-Jennings plot. We will investigate this trend further in the next step, when we add expected ranges to the plots.

Background

Levey-Jennings plots are quality control tools that help you visualize the performance of laboratory standards and quality controls over time, identifying trends and outlying data points. This can help you take corrective measures to ensure that your standards remain reliable yardsticks for your experimental data. See also: Wikipedia article on Laboratory Quality Control.

Example usage scenarios for Levey-Jennings plots:

  • If you see an outlier data point for a standard, you may investigate whether conditions were unusual on the day the data was collected (e.g., building air conditioning was not working). If the standard was not reliable on that day, other data may also be unreliable.
  • If you see a trend in the standard (as we will observe below), you may investigate whether experimental conditions are changing (e.g., a reagent is gradually degrading).
  • If standard performance changes with analyte lot, you may need to investigate the quality of the new analyte lot and potentially change the preparation or supplier of the lot.
The LabKey Luminex tool makes a set of Levey-Jennings plots available for standards for each trio of analyte, isotype and conjugate provided in the run data. Each set of plots for standards includes tabs for four different performance metrics (EC50 4PL, EC50 5PL, AUC and HighMFI). You can also generate Levey-Jennings plots for single point controls to track performance over time of controls which are not titrated.

To see which reports are available for your assay:

  • Click View QC Report > View Levey-Jennings Reports.
  • If you had checked additional well role boxes during import, such as one for single point controls, there would be additional reports listed here. See Review Well Roles for additional information.
  • Click a link to open a report.

Explore Levey-Jennings Plots for a Standard

The tutorial example includes only a single titration, so we will elect to display Levey-Jennings plots and data for the standard for the ENV2 analyte, IgA isotype and Biotin conjugate trio.

  • In the list of available reports, click Standard1.
  • In the Choose Graph Parameters box on the left side, select ENV2.
  • For Isotype, choose "IgA"
  • For Conjugate, choose "Biotin".
  • Click Apply.
    • Note: at this point in the tutorial, it is possible that you will need to add additional packages to your installation of R to support these plots. Refer to the list in Step 2: Configure R, Packages and Script, or add packages as they are requested by error messages in the UI. Retry viewing the plot after each addition until successful.
  • In the graph panel, you see a Levey-Jennings plot of EC50 - 4PL for the standard (Standard1).
  • Note the downward trend in the EC50 - 4PL, which becomes more pronounced over time and the change from Lot 437 and Lot 815.

The x-axis is labeled with the notebook numbers you entered for each run. The data points are ordered according to the acquisition date for each run, which came from the Excel file you imported for each run. Data points are spaced along the x-axis in a fixed increment, so the spacing does not reflect the actual time between runs. The data points are colored according to the analyte Lot Number.

Options just above the graph allow you to change the scale of the y-axis from linear to logarithmic, adjust the range of dates included, and select a specific network if required.

Display Levey-Jennings Plots for Other Performance Metrics

Use the tabs above the Levey-Jennings plot to see charts for:

  • EC50 - 5PL Rumi - the EC50 calculated using a 5-parameter logistic curve and the Ruminex R package
  • AUC - the area under the fluorescence intensity curve
  • HighMFI - the highest recorded flourescence intensity
  • Click on the EC50 - 5PL Rumi tab.
  • See a trend that looks quite similar to the trend for EC50 - 4PL, with points in similar positions. The exception is the value for Notebook No 05, well above the general trend.
  • Click on the AUC and HighMFI tabs to see the trends in those curves as well.

Generate PDFs for Levey-Jennings plots

If you wish, you can generate a PDF of the plot visible:

  • Click on the PDF icon in the upper right.
  • Depending on your browser settings, you may need to allow popups to run.
  • See an example PDF here.

Explore the Tracking Data Table

Below the graph area, you'll find a table that lists the values of all of the data points used in the Levey-Jennings plots above.

  • Scroll the screen down and to the right.
  • Notice the values in the last four columns.

View Levey-Jennings Plots from QC Reports

For quicker review of relevant Levey-Jennings plots without generating the full report, you can access them directly from the QC report for your titration or single point control.

  • Select (Admin) > Manage Assays, then click Luminex Assay 200.
  • Select View QC Report > View Titration QC Report.
  • Click the graph icon in the L-J Plots column of the second row (where the analyte is ENV2 we viewed earlier).
  • You can select any of the performance metrics from the dropdown. Click EC50 4PL and you can quickly review that Levey-Jennings plot.
  • Notice that the notebook number for the run whose row we selected (01 in this screencap) is shown in red along the x-axis.

Related Topics

Previous Step | Next Step (6 of 7)




Step 6: Use Guide Sets for QC


One component of validating Luminex data is to define a guide set which defines an expected range for the standard for a particular combination of analyte, isotype and conjugate. Each combination may have a different guide set. Once you apply a guide set to a run, the expected ranges are displayed in the Levey-Jennings plots. QC flags will be raised for values outside the given range. Guide sets consist of means and standard deviations for the performance metrics and may be either:
  • Run-based: calculated from an uploaded set of runs
  • Value-based: defined directly using known values, such as from historical lab data
You can define multiple guide sets of different types and choose which guide set is applied to any given run. For example, you might define a guide set based on a particular lot of analyte, and use it to check performance of that lot over time, then validate a new lot when analyte preparation or supplier has changed.

Define Guide Sets

Earlier in the Luminex Level II tutorial, we assigned five runs to each lot of analytes, so we can now create different guide sets on this data for each lot of the analyte, one run-based and one value-based. When you later select which guide set to apply, you will be able to see the comment field, so it is good practice to use that comment to provide selection guidance.

Create a Run-based Guide Set

In this tutorial example consisting of just 5 runs per lot, we use the first three runs as a guide set for the first lot. Ordinarily you would use a much larger group of runs (20-30) to establish statistically valid expected ranges for a much larger pool of data.

  • In the Luminex folder, select (Admin) > Manage Assays, then click Luminex Assay 200.
  • Open the Levey-Jennings report for the standard by selecting View QC Report > view levey-jennings reports and clicking Standard1.
  • Under Choose Graph Parameters, select the following:
    • Antigens: ENV2
    • Isotype: IgA
    • Conjugate: Biotin
    • Click Apply.
  • Above the graph, notice that the Current Guide Set box reads: "No current guide set for the selected graph parameters."
  • Click New next to this message to Create Guide Set.
    • Notice in the upper right corner of the popup, Run-based is selected by default.
    • In the All Runs panel, scroll down and click the button next to each of the Assay IDs that begin with 01, 02, and 03 to add them to the guide set.
    • Enter the Comment: "Guide Set for Lot 437"
  • Click Create.

Notice that the calculated expected ranges are shown applied to the runs you selected as part of the Guide Set. The mean is the average value, the colored bars show the calculated standard deviation. The expected range is three times the standard deviation over or under the mean.

Once you define run-based guide sets for a standard, expected ranges are calculated for all performance metrics (AUC, EC50 5PL and HighMFI), not just EC50 4PL. Switch tabs to see graphed ranges for other metrics.

Create a Value-based Guide Set

If you already have data about expected ranges and want to use these historic standard deviations and means to define your ranges, you create a value-based guide set. Here we supply some known reasonable ranges from our sample data.

  • Above the graph, click New to Create Guide Set.
  • You can only edit the most recent guide set, so you will be warned that creating this new set means you can no longer edit the guide set for Lot 437.
  • Click Yes.
  • Click Value-based under Guide Set Type.
  • Enter values as shown:
MetricMeanStd.Dev.
EC50 4PL3.620.2
EC50 5PL (Rumi)3.50.2
AUC700001000
High MFI32300200
  • Enter the Comment: "Guide Set for Lot 815"
  • Click Create.

Since this guide set is not based on any runs, you will not see expected ranges displayed in the report until it is applied.

Apply Guide Sets

For each run, you can select which guide set to apply. Once a run has a guide set applied, you may switch the association to another guide set, but may not later entirely dissociate the run from all guide sets through the user interface.

Apply Run-based Guide Set

Since we used three of our runs for the first analyte lot, we are only able to apply that guide set to the other two runs from the same lot 437.

  • At the bottom of the Levey-Jennings plot, click the checkboxes next to the the runs that begin with the digits 04 and 05.
  • Click Apply Guide Set.
  • In the popup, notice that you can see the calculated run-based thresholds listed alongside those you entered for the value-based set.
  • Select the run-based "Guide Set for Lot 437", then click Apply Thresholds.
  • In the Levey-Jennings plot, observe the range bars applied to run 04 and run 05.
  • Notice that results for both run 04 and run 05 fall outside of the expected range. We will discuss the QC flags raised by this occurrence in a future step.

Apply Value-based Guide Set

No runs were used to create this set, so we can apply it to all 5 runs that used the second analyte lot.

  • At the bottom of the Levey-Jennings plot, select the checkboxes next to runs 06-10.
  • Click Apply Guide Set.
  • In the popup, make sure that the guide set with comment Guide Set for Lot 815 is checked.
  • Click Apply Thresholds.

Notice that three of the runs from the second lot including values within our ranges, but two fall outside them.

Explore the guide sets as displayed in the graphs on the other performance metric tabs for EC50 5PL, AUC, and High MFI.

Manage Guide Sets

You can view the ranges defined by any guide set by selecting View QC Report > view guide sets and clicking Details. Note that for run-based guide sets, only the calculated range values are shown in the popup. Clicking a Graph link will navigate you to the Levey-Jennings plot where the set is defined.

Change Guide Set Associations

Select checkboxes for runs below the Levey-Jennings plot and click Apply Guide Set. You may choose from available guide sets listed. If any selected runs are used to define a run-based guide set, they may not have any guide set applied to them, and requests to do so will be ignored.

Edit Guide Sets

Only the most recently defined guide set is editable. From the Levey-Jennings plot, click Edit next to the guide set to change the values or runs which comprise it. For run based guide sets, use the plus and minus buttons for runs; for value-based guide sets, simply enter new values. Click Save when finished.

Delete Guide Sets

Over time, when new guide sets are created, you may wish to delete obsolete ones. In the case of run-based guide sets, the runs used to define them are not eligible to have other guide set ranges applied to them unless you first delete the guide set they helped define.

  • Select View QC Report > view guide sets.
  • Check the box for the obsolete guide set. To continue with the tutorial, do not delete the guide sets we just created.
  • Click (Delete).
  • For each guide set selected, you will be shown some information about it before confirming the deletion, including the set of runs which may still be using the given guide set. In this screencap, you see what the confirmation screen would look like if you attempted to delete an old run-based guide set. Value-based sets will only have user runs.
  • Click Cancel and then either Graph link to return to the Levey-Jennings plot.

When a guide set is deleted, any QC flags raised by the expected range it defined will be deleted as well.

View QC Flags

When guide sets are applied, runs whose values which fall outside expected ranges are automatically flagged for quality control. You can see these tags in the grid at the bottom of the Levey-Jennings page.

  • Look at the Standard1 Tracking Data grid below the plots.
  • Notice red highlighting is applied to any values that fall out of the applied guide set ranges.
  • Observe that QC flags have been added to the left hand column for each metric where there are out of range values on each run.

For additional information about QC flagging, including how to disable individual flags, see Luminex QC Reports and Flags.

Previous Step | Next Step (7 of 7)




Step 7: Compare Standard Curves Across Runs


Plotting standard curves for several runs together helps visualize any inconsistencies in data and curve fits between runs. The resulting overlay plot is sometimes called a curve "graveyard."

Here, we generate an overlay plot for the 4pl standard titration curves for the same data used in the previous steps (Step 5: Track Analyte Quality Over Time and Step 6: Use Guide Sets for QC).

Steps

If you navigated away after the last tutorial step, return to the Levey-Jennings plot for the ENV2 standard

  • Select (Admin) > Manage Assays in the Luminex folder.
  • Click Luminex Assay 200.
  • Select View QC Report > view levey-jennings reports.
  • Click Standard1.
  • In the Choose Graph Parameters box, select Antigen "ENV2", Isotype "IgA", Conjugate "Biotin" and click Apply.

Next, create the overlay plot:

  • Scroll down to the Standard1 Tracking Data for ENV2 - IgA Biotin table.
  • Select all rows. (Click the box at the top of the left hand column.)
  • Click View 4pl Curves to generate the overlay plot.

In the Curve Comparison popup, you can customize what is shown:

  • Y-Axis: Use the pulldown to specify what is plotted on the Y-axis. Options: FI, FI-Bkgd, FI-Bkgd-Neg
  • Scale: Choose linear or log scale.
  • Legend: Use the pulldown to specify how the legend is labeled.
  • Export to PDF - Export the overlay plot. The exported pdf includes both linear and log versions. View a sample here.
  • Close the plot when finished.
Congratulations! You have completed the Luminex Tutorial Level II.

Related Topics

Previous Step