This paper is devoted to the study of the mechanisms of interaction between uranyl ion and rutile TiO2. Among the radionuclides of interest, U(VI) can be considered as a model of the radionuclides oxo-cations. The substrate under study here is the rutile titanium dioxide (TiO2) which is an interesting candidate as a methodological solid since it can be easily found as powder and as manufactured single crystals. This material presents also a wide domain of stability as a function of pH. Then, it allows the study of the retention processes on well-defined crystallographic planes, which can lead to a better understanding of the surface reaction mechanisms. Moreover, it is well-established that the (110) crystallographic orientation is dominating the surface chemistry of the rutile powder. Therefore, the spectroscopic results obtained for the U(VI)/rutile (110) system and other relevant crystallographic orientations were used to have some insight on the nature of the uranium surface complexes formed on rutile powder. This goal was achieved by using time-resolved laser-induced fluorescence spectroscopy (TRLFS) which allows the investigation, at a molecular scale, of the nature of the reactive surface sites as well as the surface species. For rutile surfaces, oxygen atoms can be 3-fold, 2-fold (bridging oxygens), or single-fold (top oxygens) coordinated to titanium atoms. However, among these three types of surface oxygen atoms, the 3-fold coordinated ones are not reactive toward water molecules or aqueous metallic cations. This study led to conclude on the presence of two uranium(VI) surface complexes: the first one corresponds to the sorption of aquo UO22+ ion sorbed on two bridging oxygen atoms, while the second one, which is favored at higher surface coverages, corresponds to the retention of UO22+ by one bridging and one top oxygen atom. Thus, the approach presented in this paper allows the establishment of experimental constraints that have to be taken into account in the modeling of the sorption mechanisms.