PMC is capable of quantifying mixing in the presence of significant background signal

The image data for Figure 8, and Figures S7 of Redefining colocalization analysis with a novel Phasor Mixing Coefficient.
This folder contains multispectral imaging data of U2OS cells in LAMP1 was labeled with AlexaFluor(AF)-488, while TOMM20 was labeled with AF-568. Additionally, a set of cells were also stained with CellTrace-Yellow to act as a diffuse background signal.
The folder "one-color" contains the single-color control multispectral images for the two non-background signals. The file "Snap-1001.czi" refers to the single-label LAMP1; "Snap-995.czi" refers to the single-label TOMM20.
The folder "two-color" contains the two-color LAMP1+TOMM20 multispectral images. These were used as controls for the mixing analysis of the three-color samples containing CellTrace in which the diffuse background signal was removed.
The folder "one-color-plus-cellTrace" contains the single-color multispectral images for the CellTrace sample, as well as two-color samples of LAMP1+CellTrace and TOMM20+CellTrace. The files are named accordingly. This folder also contains relevant spectra for each label as measured by the Zen imaging software. The folder "three-color" contains the three-color LAMP1+TOMM20+CellTrace multispectral images. In these folders, the filenames also contain relevant imaging parameters: the laser power of 488 and 561 are 0.8 and 0.1%, respectively; the pinhole was set to 70; the gain was set to 700; and a zoom of 2x was used.
The analysis outputs are produced through the "Figure 8" section of the Jupyter notebook in the associated GitHub repository.