Measurement of oxygen consumption is a powerful and uniquely informative experimental technique. It can help identify mitochondrial mechanisms of action following pharmacologic and genetic interventions, and characterize energy metabolism in physiology and disease. The conceptual and practical benefits of respirometry have made it a frontline technique to understand how mitochondrial function can interface with—and in some cases control—cell physiology. Nonetheless, an appreciation of the complexity and challenges involved with such measurements is required to avoid common experimental and analytical pitfalls. Here we provide a practical guide to oxygen consumption measurements covering the selection of experimental models and instrumentation, as well as recommendations for the collection, interpretation and normalization of data. These guidelines are provided with the intention of aiding experimental design and enhancing the overall reputability, transparency and reliability of oxygen consumption measurements.
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A.S.D. is supported by National Institutes of Health grants R35GM138003, P30DK063491 and P50CA092131, as well as the W. M. Keck Foundation. M.J. is supported by the Novo Nordisk Research Fonden (NNF20OC0059646). We thank members of both of our laboratories for their helpful discussions during the preparation of this manuscript, as well as L. Stiles (UCLA), B. Desousa (UCSF) and A. Murphy (Cytokinetics) for their critical perspective.
The authors declare no current competing interests. A.S.D. has previously served as a paid consultant for Agilent Technologies.
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Divakaruni, A.S., Jastroch, M. A practical guide for the analysis, standardization and interpretation of oxygen consumption measurements. Nat Metab 4, 978–994 (2022). https://doi.org/10.1038/s42255-022-00619-4