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ABSTRACT 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. Access through your institution Buy or subscribe This is a preview of subscription

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BEYOND FUNCTION AND DYSFUNCTION Article 26 April 2023 MITOCHONDRIAL GENETICS, SIGNALLING AND STRESS RESPONSES Article 10 March 2025 MECHANISMS OF MITOCHONDRIAL RESPIRATORY ADAPTATION Article

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Central  Google Scholar  Download references ACKNOWLEDGEMENTS 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. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Molecular

and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA Ajit S. Divakaruni * Department of Molecular Biosciences, The

Wenner-Gren Institute, The Arrhenius Laboratories F3, Stockholm University, Stockholm, Sweden Martin Jastroch Authors * Ajit S. Divakaruni View author publications You can also search for

this author inPubMed Google Scholar * Martin Jastroch View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS A.S.D. and M.J. both contributed to

the conceptualization, preparation, writing and editing of this manuscript. CORRESPONDING AUTHOR Correspondence to Ajit S. Divakaruni. ETHICS DECLARATIONS COMPETING INTERESTS The authors

declare no current competing interests. A.S.D. has previously served as a paid consultant for Agilent Technologies. PEER REVIEW PEER REVIEW INFORMATION _Nature Metabolism_ thanks Alexander

Galkin, David Nicholls and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Primary Handling Editor: Christoph Schmitt, in collaboration with the

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ARTICLE Divakaruni, A.S., Jastroch, M. A practical guide for the analysis, standardization and interpretation of oxygen consumption measurements. _Nat Metab_ 4, 978–994 (2022).

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