In Vivo Visualization of Vasculature in Adult Zebrafish by Using High-Frequency Ultrafast Ultrasound Imaging

• Published in: IEEE transactions on biomedical engineering Vol. 66; no. 6; pp. 1742 - 1751
• Main Authors: Chang, Chao-Chuan, Chen, Pei-Yu, Huang, Hsin, Huang, Chih-Chung
• Format: Journal Article
• Language: English
• Published: United States IEEE 01.06.2019; The Institute of Electrical and Electronics Engineers, Inc. (IEEE)
• Subjects: adaptive spatiotemporal filtering; Adult zebrafish; Angiogenesis; Animals; Aorta; Blood flow; Blood vessels; Blood Vessels - diagnostic imaging; Clutter; Danio rerio; Developmental biology; Doppler effect; doppler imaging; Echocardiography; Flow velocity; Genetics; high-frequency ultrasound; Image Processing, Computer-Assisted - methods; Imaging; Imaging techniques; Mapping; Optical imaging; Optics; Regeneration; Transducers; Tumorigenesis; ultrafast ultrasound imaging; Ultrasonic imaging; Ultrasonography, Doppler - methods; Ultrasound; Vertebrates; Zebrafish
• ISSN: 0018-9294; 1558-2531
• DOI: 10.1109/TBME.2018.2878887

Objective: 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, its body loses transparency, thus making conventional optical imaging techniques difficult for imaging internal anatomy and vasculature. 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. Methods: In the present study, high-frequency micro-Doppler imaging (HFμDI) based on ultrafast ultrasound imaging was proposed for 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. Results: Blood vessels, including intersegmental vessels, parachordal vessel, dorsal longitudinal anastomotic vessel, and dorsal aorta, from the dorsal side of the zebrafish were clearly observed in two-dimensional (2-D) and 3-D HFμDI. Conclusion: 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. Significance: Compared with conventional ultrasound Doppler imaging, HFμDI exhibited superior small vessel imaging.