We report a detailed characterization of Eu3+ and Tb3+ complexes derived from a tripyridinophane macro cycle bearing three acetate side arms (H(3)tpptac). Tpptac(3-) displays an overall basicity (Sigma log K-i(H)) of 24.5, provides the formation of mononuclear ML species, and shows a good binding affinity for Ln(3+) (log K-LnL = 17.5-18.7). These complexes are also thermodynamically stable at physiological pH (pEu = 18.6, pTb = 18.0). It should be noted that the pGd value of Gd-tpptac (18.4) is only slightly lower than that of commercially available MRI contrast agents such as Gd-dota (pGd = 19.2). Moreover, a very good selectivity for these ions over the endogenous cations (log K-CuL = 14.4, log K-ZnL = 12.9, and log K-CaL = 9.3) is observed. The X-ray structure of the terbium complex shows the metal coordinated by the nine N6O3 donor set of the ligand and one inner-sphere water molecule. DFT calculations result in two Eu-tpptac structures with similar bond energies (Delta E = 0.145 eV): one structure in which the water is coordinated to the metal ion and one structure in which the water molecule is farther away from the ion, bound to the ligand with an OH-pi bond. By detailed luminescence experiments, we demonstrate that the europium complex in aqueous solution presents a hydration equilibrium between nine-coordinate, dehydrated [Eu-tpptac](0) and ten-coordinate, monohydrated [Eu-tpptac(H2O)](0) species. A similar trend is observed for the terbium complex. Despite the presence of this hydration equilibrium, the H(3)tpptac ligand sensitizes Eu3+ and Tb3+ luminescence efficiently in buffered water at physiological pH. Particularly, the terbium complex displays a long excited-state lifetime of 2.24 ms and an overall quantum yield of 33% with a brightness of 3600 M-1 cm(-1). Such features of Ln(3+) complexes of H(3)tpptac indicate that this platform appears to be particularly appealing for the further development of luminescent lanthanide labels.