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A production of heat by mitochondria is critical for maintaining bodytemperature, regulating metabolic rate and preventing oxidative damage tomitochondria and cells. Up to now mitochondrion heat production wascharacterized only by methods based on fluorescent probes which are sensitiveto environmental variations (viscosity, pH, ionic strength, quenching etc.).Herein, for the first time the heat release of isolated mitochondria wasunambiguously measured by a diamond thermometer (DT) which is absolutelyindifferent to external non-thermal parameters. We show that during totaluncoupling of transmembrane potential by CCCP application the temperature nearmitochondria rises by 4-22 {\deg}C above the ambient temperature, with anabsolute maximum of 45 {\deg}C. Such a broad temperature response may beassociated with the heterogeneity of the mitochondria themselves as well astheir aggregations in the isolated suspension. It also revealed spontaneoustemperature bursts prior to CCCP application that can reflect involvement ofsome mitochondria to ATP synthesis or membrane potential leaking to avoidreactive oxygen species hyperproduction. The used temperature sensor and thedata obtained shed light to the thermodynamics on the subcellular level.

Heat release by isolated mouse brain mitochondria detected with diamond thermometer