Dual-response fluorescent probe platform integrating two synergistic hypoxia indicators for precision imaging of hypoxic stress

Hypoxia is a pathological state caused by insufficient oxygen supply within tissues, which plays a decisive role in regulating metabolic pathways, redox homeostasis, and disease progression. Accurately visualizing the hypoxic microenvironment is crucial for understanding the pathways through which hypoxia occurs. However, numerous hypoxia probes typically only target a single analyte, thus failing to gain a deep understanding of the complex redox and enzyme disturbances of hypoxia stress. To overcome this limitation, we present a dual-response fluorescent probe platform that integrates nitroreductase (NTR) and hypochlorous acid (HOCl) as two synergistic hypoxia-related indicators to achieve multiplex imaging of hypoxic stress. In contrast to conventional single-response fluorescent probes, this platform functions as a two-input/two-output logic gate, enabling the independent or simultaneous detection of NTR and HOCl with high sensitivity and selectivity via dual-channel. This dual-indicator strategy provides a more comprehensive understanding of their respective roles during hypoxia, greatly improving the accuracy of identifying hypoxic stress. The system platform has successfully achieved efficient and accurate fluorescence imaging of hypoxic areas in cells, zebrafish, and mice. This multifunctional probe offers a promising tool for investigating hypoxia-related pathophysiology or biological processes and may facilitate the auxiliary diagnosis and therapeutic evaluation in hypoxia-associated diseases.