Spectral unmixing with CRANAD-29 for ex vivo BAT and blood. (a) Unmixed #1 represents autofluorescence signal and is corresponding to green line spectrum in (e). (b) Unmixed #1 represents NIRF signal from BAT mass and is corresponding to blue line spectrum in (e). (c) Unmixed #2 is for NIRF from blood and red line spectrum in (e). (d) Merged image of unmixed #1, #2, and #3. (e) Ex vivo unmixed spectra for autofluorescence (green), BAT mass (blue), and blood flow (red).