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This volume details comprehensive protocols and methodologies to assess mitochondrial bioenergetics and dynamics in different tissues and cells involving health and pathological states. Chapters guide readers through methods for assessment of the energy metabolism including Oxygen Consumption Rate (OCR), mitochondrial membrane potential, and measuring mitochondrial Ca2+ handling, and ROS emission. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting, and systematic reproducible protocols.