A two-photon excitation fluorescent probe enables dual-channel mapping mitochondrial superoxide anion and viscosity for evaluating multiple inflammation models

Excessive superoxide anion (O2.-) and hyperviscosity intracellular heterogeneous microenvironment are generally associated with various inflammatory disorders and therefore the biomarkers for diagnosing the related diseases while simultaneously determining the intracellular O2.- and viscosity levels in the physiological environment remains a great challenge. In this work, we developed a single fluorescent probe (CM-TBS) capable of dualchannel sensing mitochondrial O2.- species and viscosity levels, with detection limits down to 28.88 nM and 1.46 cp, respectively. Owing to such superior sensitivity and the salient property regarding dual-channel detection of two types of indicating cellular contexts, as well as the recognition specificity, second-level response agility, and two-photon excitation fluorescence (TPEF) imaging ability beneficial for achieving satisfactory imaging depth in intravital biological imaging, satisfactory results in evaluating the common inflammation diseases in clinic including acute pancreatitis, autoimmune hepatitis, interstitial cystitis were unequivocally obtained by accurately mapping the up-regulated mitochondrial O2.- level and viscosity in these cases.