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In vivo Visualization of Vasculature in Adult Zebrafish by using High-Frequency Ultrafast Ultrasound Imaging

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In vivo Visualization of Vasculature in Adult Zebrafish by High Frequency Ultrafast Ultrasound Imaging

Zebrafish has been recently considered an ideal vertebrate for studying developmental biology, genetics, particularly for modeling tumorigenesis, angiogenesis, and regeneration in vivo. However, when a zebrafish matures completely, particularly if the specimen is from a wild-type line, stripes are evident along the length of the body; such stripes reduce transparency. Optical imaging may have limitations for adult zebrafish imaging because of its low penetration ability. Acoustic wave penetration outperforms optical methods, high-frequency (>30 MHz) ultrasound (HFUS) was consequently an alternative imaging modality for adult zebrafish imaging, particularly for echocardiography However, visualizing peripheral vessels in a zebrafish by using conventional HFUS is still difficult. In the present study, high-frequency micro-Doppler imaging (HFμDI) based on ultrafast ultrasound imaging was proposed for adult zebrafish dorsal vascular mapping in vivo. HFμDI uses a 40-MHz ultrasound transducer, which is an ultrafast ultrasound imaging technology with the highest frequency available currently. Blood flow signals were extracted using an eigen-based clutter filter with different settings. Experiments were performed on an 8-month-old wild-type AB-line adult zebrafish. Blood vessels, including intersegmental vessels, parachordal vessel, dorsal longitudinal anastomotic vessel, and dorsal aorta, from the dorsal side of the adult zebrafish were clearly observed in two-dimensional (2D) and three-dimensional (3D) HFμDI. The maximum image depth of HFμDI and the minimal diameter of vessel can be detected were 4 mm and 36 μm, respectively; they were determined without any use of microbubbles. The maximum flow velocity range was approximately 3–4 mm/s on the dorsal vessels of the adult zebrafish.

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