An accurate and reliable technique used to quantify carotid plaque volume has practical importance in research and patient management. In this study, we develop and investigate a theoretical description of carotid plaque volume measurements made using three-dimensional (3D) ultrasound (US) images and compare it with experimental results. Multiple observers measured 48 3D US patient images of carotid plaque (13.2 to 544.0 mm(3)) by manual planimetry. Coefficients of variation in the measurement of plaque volume were found to decrease with increasing plaque size for both inter- (90.8 to 3.9%) and intraobserver (70.2 to 3.1%) measurements. Plaque volume measurement variability was found to increase with interslice distance (ISD), while the relative measurement accuracy remained constant for ISDs between 1.0 and 3.0 mm and then decreased. Root-mean-square (RMS) difference between our theoretical description of plaque volume measurement variance and the experimental results was 5.7%. Thus, our results support the clinical utility of measuring carotid plaque volume by manual planimetry noninvasively using 3D US.