MitoPerOx is a ratiometric fluorescent probe that can be used to assess changes in lipid peroxidation within mitochondria. It is composed of a BODIPY fluorophore conjugated via a dienyl link to a triphenylphosphonium cation component that drives its accumulation in mitochondria. MitoPerOx displays an excitation maximum of 495 nm and exhibits a shift in emission maxima from 590 to 520 nm upon mitochondrial lipid peroxidation, enabling determination of ratiometric measurements of lipid peroxidation in live cells.