International audienceBackground/Purpose The ASCLEPIOS system developed by the M2D+ team of the Le2i laboratory (Université de Bourgogne, France) allows determination of a skin reflectance spectrum over the visible wavelength range in each pixel of a 2D image, thereby generating a hyperspectral (3D) cube. Reflectance spectra mainly result from the reflectance of two skin chromophores, epidermal melanin and dermal haemoglobin. A source separation method was applied on the mixed reflectance spectra, resulting in two component spectra for melanin and haemoglobin, respectively. We also obtained through this process quantification of each chromophore in each pixel of a 2D skin image. The accuracy of the pure source spectra obtained was validated...