Facile construction of highly luminescent and biocompatible gold nanoclusters by shell rigidification for two-photon pH-edited cytoplasmic and in vivo imaging

Gold nanoclusters (AuNCs), as a novel fluorescent material, have been extensively explored and developed for bioimaging because of their attractive advantages such as ultrasmall size, low toxicity and exceptional two-photon excitation properties. However, it still remains a challenge to produce water-soluble, biocompatible and ultrabright AuNCs. Herein, we report on a novel one-pot synthesis of highly luminescent and biocompatible AuNCs by using polyvinyl pyrrolidone (PVP), a water-soluble polymer, to rigidify the primary stabilizing layer (shell) that is composed of 6-aza-2-thiothymine (ATT) ligands bound to the particle. Such shell-rigidification resulted in a significant enhancement of the fluorescence efficiency, reaching a quantum yield of 39% under the best conditions, about 35-fold increase from the intrinsically weak fluorescence of the AuNCs stabilized by only ATT. The fluorescence enhancement mechanism was systematically characterized, and the results indicate that PVP coating rigidifies the ATT ligand shell through steric hindrance and reduces the nonradiative relaxation of the excited states. The biocompatible PVP-AuNCs were further examined for two-photon cellular and sentinel lymph node (SLN) bioimaging, and we observed pH-dependent cytoplasmic images and intense green fluorescence in SLN and lymphatic vessels.