Abstract

Mitochondrion is one of significant organelles inside cells because it serves as a hub for energy management and intracellular signaling. Internal/external damages on mitochondria would lead to mitochondrial stresses with the malfunctions, accompanying with the changes of morphological structure and abnormal local environments (pH values). Mitophagy is capable of degradation of damaged mitochondrial segments to restore its normal metabolism, dynamics, and biogenesis. The dynamic structural visualization and pH quantification can be helpful for the understanding of mitochondrial functions as well as the diagnosis of disorders linking with this process. In this work, we use confocal laser scanning microscopy, STED super-resolution nanoscopy and fluorescence lifetime imaging microscopy, in conjunction with a mitochondrial probe to image the dynamic changes in the mitochondrial morphology and microenvironmental pH values during mitophagy in live cells, in particular, the structural changes of mitochondrial cristae beyond optical diffraction can be distinguished by STED nanoscopy with/without treatment by CCCP, which will provide a new view for the diagnosis and personalized treatment of mitochondrial dysfunction-related diseases.

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