Ultrasensitive triple-mode Nile red-L64 niosome nanothermometers for real-time cellular thermogenesis mapping

with astounding temperature resolution of less than 0.001 °C, under two-photon fluorescence imaging. NR-L64 demonstrates remarkable photostability for over an hour and maintains a linear thermal response in the range of 37-39 °C. The nanothermometers are successful in monitoring real-time glucose-induced thermogenesis in live FaDu cells, by detecting a sequential temperature increase of 0.2 °C with subcellular resolution using confocal and two-photon fluorescence microscopy. Cross-validation between intensity and lifetime imaging confirms reliable thermal quantification across the linear range. These findings establish NR-L64 niosomes as versatile, minimally invasive tools for real-time cellular thermometry with broad applications in metabolic studies and precision therapeutic monitoring.