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We are developing a hyperspectral imaging system aimed at imaging fluorescent molecules in two different contexts: In vivo 3-D molecular imaging of mice and multi-fluorophore imaging of 2-D tissue samples. The main concept is to add high resolution spectral information to the intensity data to improve imaging capabilities. The system is based on an imaging spectrograph, ImSpector, coupled to a CCD camera. The system has been characterized and calibrated. Spectral imaging of tissue samples stained with fluorescent dyes has been performed to validate the principle and the 2-D imaging capabilities. Near infrared 3-D in vivo fluorescence tomography is based on the intensity and spatial distribution of the fluorescent light reaching the surface of the animal as a function of the localized external excitation position. Based on the fact that the absorption coefficient of tissue is wavelength dependent, the measured spectrum of the fluorescent light will provide additional information, thus allowing an improvement of the 3-D reconstruction. By using solely spectral data we have demonstrated the capability of determining the 3-D position of a localized source within a red meat phantom.