Method for reconstructing a three-dimensional fluorescence-optical-tomography image for examining an object of plate shape having a first face and an opposite second face and containing fluorophores, the method comprising: - a first step of sequential illumination of the first face of the object with light (λex) for exciting the fluorophores from a plurality of illumination points, the first illumination step comprising a succession of phases of illumination from an illumination point associated with each phase, and - a first step of sequential acquisition of a first series of images, an image of said series being formed, in each phase of illumination of the first face, by a detector comprising a plurality of detection points that simultaneously detect light (λex, λem) emitted via the second face of the object, the method being such that the density of the illumination points is lower than the density of the detection points, the method then comprising: - a second step of sequential illumination of the second face of the object with light (λex) for exciting the fluorophores, - a second step of sequential acquisition of a second series of images by detecting light (λex, λem) emitted via the first face of the object, and - reconstruction of the three-dimensional image of the distribution of the fluorophores in the object by way of the first and second series of images, said reconstruction being achieved by establishing two linear systems of equations that are established using two sets of matrices of Green's functions corresponding to the first and second acquisition of series of images, respectively, the two linear systems of equations being combined to be solved; the method being characterized in that the number and distribution of the illumination points and of the detection points differs between the first step of sequential illumination and acquisition and the second step of sequential illumination and acquisition. - a first step of sequential illumination of the first face of the object with light (λex) for exciting the fluorophores from a plurality of illumination points, the first illumination step comprising a succession of phases of illumination from an illumination point associated with each phase, and - a first step of sequential acquisition of a first series of images, an image of said series being formed, in each phase of illumination of the first face, by a detector comprising a plurality of detection points that simultaneously detect light (λex, λem) emitted via the second face of the object, the method being such that the density of the illumination points is lower than the density of the detection points, the method then comprising: - a second step of sequential illumination of the second face of the object with light (λex) for exciting the fluorophores, - a second step of sequential acquisition of a second series of images by detecting light (λex, λem) emitted via the first face of the object, and - reconstruction of the three-dimensional image of the distribution of the fluorophores in the object by way of the first and second series of images, said reconstruction being achieved by establishing two linear systems of equations that are established using two sets of matrices of Green's functions corresponding to the first and second acquisition of series of images, respectively, the two linear systems of equations being combined to be solved; the method being characterized in that the number and distribution of the illumination points and of the detection points differs between the first step of sequential illumination and acquisition and the second step of sequential illumination and acquisition. - a second step of sequential illumination of the second face of the object with light (λex) for exciting the fluorophores, - a second step of sequential acquisition of a second series of images by detecting light (λex, λem) emitted via the first face of the object, and - reconstruction of the three-dimensional image of the distribution of the fluorophores in the object by way of the first and second series of images, said reconstruction being achieved by establishing two linear systems of equations that are established using two sets of matrices of Green's functions corresponding to the first and second acquisition of series of images, respectively, the two linear systems of equations being combined to be solved; the method being characterized in that the number and distribution of the illumination points and of the detection points differs between the first step of sequential illumination and acquisition and the second step of sequential illumination and acquisition.