Fluorescence-Based Quantification of Mitochondrial Damage in Human Airway Smooth Muscle Cells

Dual-fluorescence labeling reveals that untreated human airway smooth muscle cells exhibit ~10% overlap between intact and damaged mitochondria.

• Mitochondrial damage is indicated by loss of mitochondrial membrane potential, which triggers the process of mitophagy.
• Existing tools for assessing mitochondrial damage often provide limited or semiquantitative measures.
• The developed imaging-based assay effectively quantifies intact versus damaged mitochondria in human airway smooth muscle cells.
• Treatment with the mitochondrial uncoupler FCCP leads to significant mitochondrial depolarization, reducing fluorescence overlap and increasing mitochondrial fragmentation.
• The redox-sensitive reporter pMitoTimer shows a shift in fluorescence from green to red, suggesting increased oxidative stress and turnover of damaged mitochondria.

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