FlowJo will do the entire unmixing process in once click with this selection.
The one touch approach uses spectral unmixing, auto-spill, and our standard clean-up gate creation to perform all of the steps needed to create a spillover matrix. It will automatically use all detectors, but importantly, if you have a traditional compensation assay, in which there are an equivalent number of detectors and parameters, spectral unmixing and traditional compensation are equivalent.
NOTE: One touch does not allow for any user modification of the clean-up gates or parameter assignments, so if only works well on high-quality single stain controls.
Use
To perform One-touch unmixing:
- Select the Compensation context, highlighted in Figure 1
- Click on the specific set of single stain controls that you would like to use, found within the Compensation Data group
- Click the Auto button, also highlighted in Figure 1
Figure 1 The steps for performing One touch unmixing
The software will display an in-progress screen while calculating. Calculation time will depend on the complexity of the assay and can take several minutes.
Figure 2 Creating a matrix
When complete a new matrix, called simply 'Matrix', will appear within the list of Compensation matrices. The matrix will be selected, as shown in Figure 3, and the outcome of the calculation will be shown in the primary pane. The matrix then acts as any other matrix, including that it can be applied to any sample with the matching staining panel or previewed on any sample.
Figure 3 Completed One touch unmixing
For more information on unmixing or compensation see:
- Compensation / Unmixing in FlowJo
- The Compensation Wizard
- AutoSpill
- Matrix management