Imaging the Dynamics of Mitophagy in Live Cells

Abstract

Investigating the precise spatiotemporal dynamics of mitophagy can provide insights into how mitochondrial quality control is regulated in different tissues and organisms. Here, we outline live imaging assays to quantitatively assess mitophagy dynamics in real time. This protocol describes both chemical and optogenetic techniques to induce mitochondrial damage with high spatial and temporal control. Using these assays, mitochondria can be tracked from before they sustain damage up to their engulfment by autophagosomes and acidification by lysosomes.

Keywords: Autophagy; LC3; Lysosome; Mitochondria; Mitophagy; OPTN; Parkin; Ubiquitin.