Bovine serum albumin protected Cd₈ nanoclusters as efficient two-photon absorbers for near-infrared excited fluorescence imaging of intracellular pH

Metal nanocluster-based fluorescent sensors have made great progress in the field of analytical chemistry, but their practical applications in biological sensing were limited by poor stability, short wavelength excitation and low quantum yield. It is of great significance to develop NIR-excited metal nanocluster-based fluorescence sensors with high quantum yield. In contrast to noble metal nanoclusters (e.g., Au, Ag, and Pt), stable and affordable fluorescent metal nanoclusters have received relatively less research attention, with the exception for Cu nanoclusters. Herein, we developed a rapid one-pot route to synthesize cadmium nanoclusters employing bovine serum albumin (BSA) as both reducing and templating agent. The obtained Cd₈NCs exhibited bright fluorescence emission with quantum yield of 13.8%. Of particular interest, the Cd₈NCs showed remarkable capability as two-photon absorbers, with a two-photon absorption (TPA) cross section up to 1588 GM (excited at 730 nm). The TPA cross section was higher than that of BSA-protected AuNCs, and even exceeded that of conventional dyes by two orders of magnitude. In addition, the Cd₈NCs exhibited pH-sensitive photoluminescence, and the pH-responsive mechanism was revealed by time-resolved fluorescence spectroscopy, transient absorption spectroscopy, etc. Benefiting from protein coating, Cd₈NCs showed excellent biocompatibility and were successfully utilized as two-photon excited fluorescence sensors for intracellular pH.