Nitrogen-Rich D-π-A Structural Carbon Quantum Dots with a Bright Two-Photon Fluorescence for Deep-Tissue Imaging.

Two-photon fluorescent (TPF) probes, which allow imaging of biological events in a high spatiotemporal resolution, are in great demand. Recently, carbon quantum dots (CQDs) have emerged as a promising class of TPF probes. Unfortunately, the use of the existing CQDs has been limited by their weak TPF capacities. Herein, we report the first facile and large-scale synthesis of nitrogen-rich CQDs (NRCQDs) based on a donor-π-acceptor (D-π-A) strategy. The resulting NRCQDs demonstrated a tremendous TPF capacity with a two-photon absorption cross section (TPACS) and quantum yield (QY) up to 61 200 Göppert-Mayer (GM) units and 63%, respectively, which is greater than those that could be achieved by the existing TPF carbon probes. Structural and optical analyses of NRCQDs revealed that the great TPF capacity is contributed by the nitrogen-rich D-π-A structure as well as the high crystallinity, large plane, rigid, graphitic nitrogen-doped π-conjugated system. We further demonstrated that NRCQDs allow imaging of live cells as well as live liver tissues at depths of up to 440 μm. Our results suggest NRCQDs as a robust TPF probe that can be potentially used for a variety of biological applications.