Membrane-Activated Fluorescent Probe for High-Fidelity Imaging of Mitochondrial Membrane Potential

Abstract

Mitochondrial membrane potential (ΔΨ_m) is a key indicator of cell health or injury due to its vital roles in adenosine 5'-triphosphate synthesis. Thus, monitoring ΔΨ_m is of great significance for the assessment of cell status, diagnosis of diseases, and medicament screening. Cationic fluorescent probes suffer from severe photobleaching or false positive signals due to the luminescence of the probe on non-mitochondria. Herein, we report a lipophilic cationic fluorescent probe [1-methyl-2-(4-(1,2,2-triphenylvinyl)styryl)-β-naphthothiazol-1-ium trifluoromethanesulfonate (<b>TPE-NT</b>)] with the features of aggregation-induced emission and intramolecular charge transfer for imaging ΔΨ_m in live cells. <b>TPE-NT</b> is enriched on the surface of the mitochondrial inner membrane due to the negative ΔΨ_m, and its fluorescence is activated in the high-viscosity microenvironment. The false positive signals of emission from <b>TPE-NT</b> on non-mitochondria are therefore effectively eliminated. Moreover, <b>TPE-NT</b> exhibits a Stokes shift of >200 nm, near-infrared (∼675 nm) emission, excellent photostability, and low cytotoxicity, which facilitate real-time imaging in live cells. Cell imaging confirmed that the probe can rapidly and reliably report mitochondrial depolarization (decrement of ΔΨ_m) during cell damage caused by CCCP and H₂O₂ as well as mitochondrial polarization (increment of ΔΨ_m) by oligomycin. Furthermore, the probe successfully detected the reduction of ΔΨ_m in these cell models of hypoxia, heat damage, acidification, aging, inflammation, mitophagy, and apoptosis caused by hypoxia, heatstroke, lactate/pyruvate, doxorubicin, lipopolysaccharide, rapamycin, monensin, and nystatin, respectively.