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A FRET-based carbon nanodot (CDot) drug delivery platform has been developed. These CDots offer excellent biocompatibility, stable fluorescence, and efficient FRET between CDots and the attached fluorescent drug molecules, such as doxorubicin, enabling enhanced drug delivery, convenient cell imaging, and real-time monitoring of drug release. Moreover, the FRET-based two-photon imaging and drug tracking in deep tissues are also demonstrated.