Unimolecular dual-reporter probes for early kidney injury diagnosis through synchronous <i>in situ</i> imaging and portable urinalysis

Abstract Early detection of acute kidney injury (AKI) is critical for effective intervention and improved survival, yet it remains challenging due to the lack of practical tools that sensitively capture subtle renal dysfunction. Here, we report a unimolecular dual-reporter probe, DHU-AKI-3, featuring activatable crosstalk-free multiplexed signal output for precise diagnosis of early AKI by integrating in situ imaging and in vitro portable urinalysis. Following systemic administration, DHU-AKI-3 is selectively activated in the injured kidney, producing a long-retained photoacoustic signal for real-time in situ imaging ahead of the clinical diagnostic window. Concurrently, a near-infrared fluorescent signal is rapidly released and excreted into urine, enabling smartphone-assisted colorimetric analysis for noninvasive, point-of-care testing. The mutual validation of crosstalk-free in situ and in vitro signals guaranteed the diagnostic accuracy. This study underscores the potential of unimolecular dual-reporter probe for directly visualizing early-stage AKI in urine, meeting the pressing demand for point-of-care testing in AKI management.