Integrating genetically encoded fluorescent sensors to elucidate the spatiotemporal choreography of necrosis by sodium overload

• Genetically encoded fluorescent sensors enable real-time, subcellular monitoring of ion fluxes and metabolic changes during NECSO. • NC1-induced TRPM4 activation leads to rapid potassium loss and sodium overload, triggering a distinct necrotic pathway. • The study reveals NADH accumulation and ATP depletion in mitochondria and cytosol, alongside reduced mitochondrial oxidative stress. • This methodological framework offers a powerful tool for studying metabolic and redox dynamics in various physiological and toxicological contexts.