Fluorescence-Based Quantification of Mitochondrial Membrane Potential and Superoxide Levels Using Live Imaging in HeLa Cells

Mohammad Fazli; Chantell  S. Evans

doi:10.3791/65304

/

Fluorescence-Based Quantification of Mitochondrial Membrane Potential and Superoxide Levels Using Live Imaging in HeLa Cells

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Biology

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JoVE Journal Biology

Fluorescence-Based Quantification of Mitochondrial Membrane Potential and Superoxide Levels Using Live Imaging in HeLa Cells

7,204 Views

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06:57 min

•

May 12, 2023

DOI:

10.3791/65304-v

DOI:

10.3791/65304-v

•

06:57 min

May 12, 2023

•

37 Views

Mohammad Fazli¹, Chantell S. Evans¹

¹Department of Cell Biology,Duke University Medical Center

Transcript

This research focuses on mitochondria dysregulation in neurodegenerative diseases. We believe that this research can be used to understand potential causes for disease onset that could lead to therapeutics combating neurodegenerative diseases like Parkinson’s disease and ALS. Techniques such as super resolution microscopy like STED and SIM or expansion microscopy have improved the ability to accurately understand protein distribution within individual organelles and mitochondria distribution throughout the cell.

Results of this technique can be used as a starting point to study the effect of Parkinson’s disease linked mutations on mitochondrial turnover and begin to understand the importance of regulating reactive oxygen species levels and mitochondrial membrane potential to maintain neuronal health. Our laboratory aims to mechanistically characterize individual mitochondrial quality control pathways to understand the interplay between these pathways. By having insights into pathway dynamics, one can understand how mitochondria are maintained and how mitochondrial dysregulation contributes to neurodegenerative disease onset.

Summary

This technique describes an effective workflow to visualize and quantitatively measure mitochondrial membrane potential and superoxide levels within HeLa cells using fluorescence-based live imaging.