International audienceFluorescence optical imaging use one or several (in multiplexing) injected fluorescent markers which specifically bind to targeted compounds. Near infrared light illuminates the region of interest and the emitted fluorescence is analyzed to localize fluorescence sources. A spectroscopic approach and a separation source method (Nonnegative matrix factorization) are explored to separate di fferent fluorescence sources and remove the unwanted biological tissues autofluorescence. We present unmixing results on overlapping spectra of interest, and show that autofluorescence removal improves Fluorescent Diffuse Optical Tomograph
