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Freehand three-dimensional ultrasound imaging of carotid artery was performed by motion tracking technology.The probe positions in 3D space coordinates were calibrated at high spatial and temporal resolutions.Human carotid artery 3D ultrasound image was obtained during a freehand ultrasound examination.Ultrasound imaging has been extensively used for determining the severity of carotid atherosclerotic stenosis. In particular, the morphological characterization of carotid plaques can be performed for risk stratification of patients. However, using 2D ultrasound imaging for detecting morphological changes in plaques has several limitations. Due to the scan was performed on a single longitudinal cross-section, the selected 2D image is difficult to represent the entire morphology and volume of plaque and vessel lumen. In addition, the precise positions of 2D ultrasound images highly depend on the radiologists' experience, it makes the serial long-term exams of anti-atherosclerotic therapies are difficult to relocate the same corresponding planes by using 2D B-mode images. This has led to the recent development of three-dimensional (3D) ultrasound imaging, which offers improved visualization and quantification of complex morphologies of carotid plaques. In the present study, a freehand 3D ultrasound imaging technique based on optical motion tracking technology is proposed. Unlike other optical tracking systems, the marker is a small rigid body that is attached to the ultrasound probe and is tracked by eight high-performance digital cameras. The probe positions in 3D space coordinates are then calibrated at spatial and temporal resolutions of 10 μm and 0.01 s, respectively. The image segmentation procedure involves Otsu's and the active contour model algorithms and accurately detects the contours of the carotid arteries. The proposed imaging technique was verified using normal artery and atherosclerotic stenosis phantoms. Human experiments involving freehand scanning of the carotid artery of a volunteer were also performed. The results indicated that compared with manual segmentation, the lowest percentage errors of the proposed segmentation procedure were 7.8% and 9.1% for the external and internal carotid arteries, respectively. Finally, the effect of handshaking was calibrated using the optical tracking system for reconstructing a 3D image.