A new reconstruction algorithm for fluorescence diffuse optical tomography of biological tissues is proposed. The radiative transport equation in the frequency domain is used to model light propagation. The adjoint method studied in this work provides an efficient way for solving the inverse problem. The methodology is applied to a 2D tissue-like phantom subjected to a collimated laser beam. Indocyanine Green is used as fluorophore. Reconstructed images of the spatial fluorophore absorption distribution is assessed taking into account the residual fluorescence in the medium. We show that illuminating the tissue surface from a collimated centered direction near the inclusion gaves a better reconstruction quality. Two closely positioned inclusions can be accurately localized. Additionally, their fluorophore absorption coefficients can be quantified. However, the algorithm failes to reconstruct smaller or deeper inclusions. This is due to light attenuation in the medium. Reconstructions with noisy data are also achieved with a reasonable accuracy. Keywords fluorescence molecular imaging, radiative transport equation, modified finite volume method, frequency domain, inverse fluorescent source problem, Lagrangian formulation, adjoint method, biological tissue. Nomenclature c speed of light in vacuum (= 2.99793 10 8), m s −1 C concentration, M = mol cm −3 d measured fluorescent light intensity g anisotropy factor of the Henyey-Greenstein phase function * adjoint operator em emission field ex excitation field