Accurate Monitoring and Multiple Evaluations of Mitophagy by a Versatile Two-Photon Fluorescent Probe.

Mitophagy plays a critical role in regulating and maintaining cellular functions, particularly regulating the quantity and quality of mitochondria. In this research, a multifunctional two-photon fluorescent probe Mito-PV with improved mitochondria-anchored ability was designed. The proposed probe can track the fluctuation of polarity and viscosity in mitochondria simultaneously with two well-distinguished emissions. It can also precisely visualize the change in mitochondrial morphology (including mitochondrial form factor and length). The real-time and accurate monitoring of mitophagy under two-photon excitation was successfully achieved by utilizing probe Mito-PV through supervising the alterations of diverse mitophagy-related parameters (including colocalization coefficient, polarity, viscosity, and mitochondrial morphology). In addition, probe Mito-PV can be applied to evaluate drug bpV(phen) as an effective mitophagy inhibitor. Therefore, our work may provide a more efficient and reliable method for precisely monitoring mitophagy from multiple evaluations.