Fig. 1

Multispectral imaging and data analysis workflow. Experimental workflow, imaging protocols and data analysis pipelines are similar for mouse and human tissues, but they include optimized conditions for sample preparation (see “Implementation”). Multispectral imaging is performed using a Zeiss LSM780 confocal microscope to acquire lambda stacks. We show a single z-plane across the electromagnetic spectrum pseudo-colored by wavelength. Spectral signatures are extracted in MATLAB using the vertex component analysis. Finally, spectral unmixing is performed using different algorithms to separate signals for individual fluorophores