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Phase-change droplets (PCDs) are the novel ultrasound-responsive drug carriers composed of volatile perfluoropentanestablized by a shell of phospholipids.PCDs can undergo liquid-to-gas transition upon ultrasound exposure to turn into gas bubbles in a few microseconds, during which the mechanical forces are expected to stimulate bioeffects to ambient tissues and trigger drug liberation from the gas-liquid interfaces.These features provide great diagnostic and therapeutic usefulness of PCDs in ultrasound controlled drug delivery.In this study,we aimed to investigate the transient dynamics of drug release from PCDs and the resulting drug/tissue interactions over different spatial and temporal scales.By developing multi-color integrated high-speed fluorescence microscope system,we performed real-time observations to explore the effects of the drug release from PCDs on tumor microenvironments such as tumor angiogenesis and progression.The obtained results were correlated with the results of animal studies using mouse window chamber models and intravital microscopy, providing valuable information for validation and optimization of the therapeutic efficacy.The outcomes of this stud this study are to promote the research and development of ultrasound drug delivery using PCDs and provide advantageous tools for studies of other drug delivery techniques.