LAUSR

Significance The temperature dynamics inside living cells play a critical role in maintaining the cells’ homeostasis or driving the cells into dysfunction status. Here, we developed temperature-sensitive LysoDots and MitoDots to monitor the in situ temperature dynamics with specific organelle-targeting strategies. We demonstrated both LysoDots and MitoDots for simultaneously mapping the in situ lysosomal and mitochondrial thermal dynamics and thermal transitions in living cells with high sensitivity. Our observations indicate different metabolic pathways and thermal transitions between lysosomes and mitochondria. The nanothermometry probes provide a powerful tool for multimodality functional imaging of subcellular organelles and interactions with high spatial, temporal, and thermal dynamics resolutions.