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Cells were washed with PBS and imaged in RMPI 1640 medium without phenol red using the confocal settings of a Leica Stellaris 8 STED FALCON in an 86X objective and a 10% power 405 nm excitation laser and emission ranges of 415-465 nm and 470-520 nm for Laurdan, plus a 5% power 647 nm excitation laser for SNAP-Surface Alexa Fluor 647

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2.2 Mitochondrial dysfunction and oxidative stress Mitochondrial complex I deficiency is consistently observed in PD, leading to impaired ATP synthesis and overproduction of reactive oxygen species (ROS) (Chang and Chen, 2020