Related Experiment Video
Updated: Feb 28, 2026
Author Spotlight: Oxygen-Independent Assays to Measure Mitochondrial Function in Mammals
Published on: May 19, 2023
Oxidative phosphorylation measurement in cell lines and tissues
Nicole J Van Bergen1, Rachel E Blake1, Jonathan G Crowston1
1Centre for Eye Research Australia, 32 Gisborne St, East Melbourne, VIC 3002, Australia; Department of Ophthalmology, University of Melbourne, 32 Gisborne St, East Melbourne, VIC 3002, Australia; Royal Victorian Eye and Ear Hospital, 32 Gisborne St, East Melbourne, VIC 3002, Australia.
Mitochondrial oxidative phosphorylation (OXPHOS) dysfunction links to many diseases. This study consolidates methods for analyzing OXPHOS in cells and tissues, aiding researchers in disease research.
Area of Science:
- Biochemistry and Molecular Biology
- Cell Biology
- Pathology
Background:
- Mitochondrial oxidative phosphorylation (OXPHOS) dysfunction is increasingly linked to diverse diseases, including neurodegeneration and cancer.
- Analyzing OXPHOS is crucial for understanding these pathologies, but requires standardized and accessible methods.
Purpose of the Study:
- To consolidate and present optimized protocols for assessing mitochondrial function.
- To provide a comprehensive guide for measuring ATP synthesis, respiration, and OXPHOS enzymology.
- To assist researchers, particularly new users, in overcoming common pitfalls in mitochondrial analysis.
Main Methods:
- Whole-cell ATP synthesis and respiration assays.
- Mitochondrial isolation from cultured cells and tissue samples.
- Enzymology assays for specific OXPHO S complexes.
- Detailed protocols and optimization guidance provided.
Main Results:
- Consolidated and validated methods for comprehensive OXPHOS analysis.
- Identification of common pitfalls and strategies for optimization.
- Accessible protocols suitable for various research settings.
Conclusions:
- Standardized methods for OXPHOS analysis are essential for advancing disease research.
- This work provides a valuable resource for researchers studying mitochondrial dysfunction.
- Optimized protocols facilitate accurate assessment of mitochondrial health in diverse biological samples.
