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Live-Cell Imaging

Live Imaging: Mitophagy

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To assess mitophagy in real time, I performed live-cell imaging of neurons treated with Parkin agonists. These assays tracked mitochondrial clearance and Parkin translocation to depolarized mitochondria, demonstrating that pharmacological activation of Parkin enhances mitophagy flux. This dynamic approach provided direct evidence that Parkin agonists restore mitochondrial quality control in dopaminergic neurons (Manuscript in preparation).

Dopaminergic neurons treated with Parkin activators, were stained by lysotracker (green) and mitotracker (red) . Colocalization of green and red shows mitophagy in action. 

Lysosomal integrity was assessed using LysoSensor and LysoTracker staining in dopaminergic neurons under different treatment conditions.

Live-Imaging: Lysosomal pH

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Lysosomal integrity was assessed using LysoSensor and LysoTracker staining in dopaminergic neurons under different treatment conditions. Neurons treated with IFN-γ or with the dual-hit paradigm (PFF+IFN-γ) showed lysosomes with higher pH values (measured by LysoSensor) compared to control and PFF-only groups, indicating impaired acidification. Complementary LysoTracker staining revealed a marked reduction in the number of lysosome-positive puncta in IFN-γ– and dual-hit–treated neurons, further confirming disrupted lysosomal function

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