Method for reconstructing by optical tomography the distribution of fluorophores (f) in a nonuniform scattering object, including: - illuminating the object in continuous light with at least one light source; - measuring the light intensity (φ m) received by at least one detector at a fluorescence wavelength; - and solving, at each point (r) of a mesh of a volume at least partially encompassing the object to be observed, an equation expressing the relationship between the measured light intensity (φ m), a parameter ( β ( r f)) representative of the properties of the fluorophores (f) and first and second transfer functions ( F ( k ex, r s, r f), F ( k ex, r s, r a)) that are representative of the transfer of energy between the source and a fluorophore (f), and between a fluorophore and a detector (d), respectively, the first and second transfer functions depending on a wave number (k ex) at the excitation wavelength (λ ex) and a wave number (k fl) at the fluorescence wavelength (λ fl), respectively, the method being characterized in that it includes: - determining, at the excitation wavelength (λ ex), a three-dimensional map of an attenuation variable (k' ex (r)) representative of absorption and scattering nonuniformities of the object, the attenuation variable being dependent on the wave number and a spatial variation in a scattering coefficient at each point of the mesh; - determining a three-dimensional map of the attenuation variable (k' fl (r)) at the fluorescence wavelength (λ fl); and - reconstructing the distribution of the fluorophores using, in said first and second transfer functions, the attenuation variable (k' ex (r)) at the excitation wavelength (λ ex) and the attenuation variable (k' fl (r)) at the fluorescence wavelength (λ fl), respectively. - illuminating the object in continuous light with at least one light source; - measuring the light intensity (φ m) received by at least one detector at a fluorescence wavelength; - and solving, at each point (r) of a mesh of a volume at least partially encompassing the object to be observed, an equation expressing the relationship between the measured light intensity (φ m), a parameter ( β ( r f)) representative of the properties of the fluorophores (f) and first and second transfer functions ( F ( k ex, r s, r f), F ( k ex, r s, r a)) that are representative of the transfer of energy between the source and a fluorophore (f), and between a fluorophore and a detector (d), respectively, the first and second transfer functions depending on a wave number (k ex) at the excitation wavelength (λ ex) and a wave number (k fl) at the fluorescence wavelength (λ fl), respectively, - determining, at the excitation wavelength (λ ex), a three-dimensional map of an attenuation variable (k' ex (r)) representative of absorption and scattering nonuniformities of the object, the attenuation variable being dependent on the wave number and a spatial variation in a scattering coefficient at each point of the mesh; - determining a three-dimensional map of the attenuation variable (k' fl (r)) at the fluorescence wavelength (λ fl); and - reconstructing the distribution of the fluorophores using, in said first and second transfer functions, the attenuation variable (k' ex (r)) at the excitation wavelength (λ ex) and the attenuation variable (k' fl (r)) at the fluorescence wavelength (λ fl), respectively.