Organelle-SpecificQuantum Thermometry Using FluorescentNanodiamonds: Insights into Cellular Metabolic Thermodynamics

Intracellular thermometry is a powerful method for studying biological thermodynamics across various physiological contexts. In this study, we present an organelle-specific quantum thermometry utilizing nitrogen-vacancy (NV) centers in fluorescent nanodiamonds (FNDs) to obtain precise temperature measurements at the subcellular level. By conjugating antibodies, FNDs were selectively targeted to mitochondria, nuclei, and cell membranes in living fibroblasts, enabling real-time monitoring of temperature changes during adenosine triphosphate (ATP) synthesis and inhibition. The system integrates advanced bioconjugation and quantum sensing methodologies, thereby overcoming challenges, such as photobleaching and limited spatial resolution. Notably, mitochondria-targeted FNDs revealed significant temperature increases, revealing mitochondria as the primary site of thermogenesis during ATP inhibition. These findings establish a robust framework for investigating metabolic thermodynamics and offer a powerful tool for exploring the thermal regulation of cellular processes.