Precise and differential profiling of the dynamic correlations and pathophysiological implications of multiplex biological mediators with deep penetration and highly programmed precision remain critical challenges in clinics. Here we present an innovative strategy by tailoring a powerful multispectral optoacoustic tomography (MSOT) technique with a photon-upconverting nanoprobe (UCN) for simultaneous visualization of diversely endogenous redox biomarkers with excellent spatiotemporal resolution in living conditions. Upon incorporating two specific radicals-sensitive NIR cyanine fluorophores onto UCNs surface, such nanoprobes can orthogonally respond to disparate oxidative and nitrosative stimulation, and generate spectrally opposite optoacoustic signal variations, which thus achieves compelling superiorities for reversed ratiometric tracking of multiple radicals under dual independent wavelength channels, and significantly, for precise validating of their complex dynamics and correlations with redox-mediated pathophysiological procession in vivo.

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