Fluorescence imaging in centimeter-deep tissues with high resolution is highly desirable for many biomedical applications. Recently, we have developed a new imaging modality, ultrasound-switchable fluorescence (USF) imaging, for achieving this goal. In our previous work, we successfully achieved USF imaging with several types of USF contrast agents and imaging systems. In this study, we introduced a new USF imaging system: an intensified charge-coupled device (ICCD) camera-based, time-domain USF imaging system. We demonstrated the principle of time-domain USF imaging by using two USF contrast agents. With a series of USF imaging experiments, we demonstrated the tradeoffs among different experimental parameters (i.e., data acquisition time, including CCD camera recording time and intensifier gate delay; focused ultrasound (FU) power; and imaging depth) and the image qualities (i.e., signal-to-noise ratio, spatial resolution, and temporal resolution). In this study, we also discussed several imaging strategies for achieving a high-quality USF image via this time-domain system.
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