Intravascular ultrasound (IVUS), which depicts both lumen and plaque, offers the potential to improve on the limitations of angiography for the assessment of the natural history of atherosclerosis and progression or regression of the disease. To facilitate measurements and increase the reproducibility of quantitative IVUS analyses, a computerized contour detection system was developed that detects both the luminal and external vessel boundaries in 3-dimensional sets of IVUS images. To validate this system, atherosclerotic human coronary segments (n = 13) with an area obstruction > or = 40% (40% to 61%) were studied in vitro by IVUS. The computerized IVUS measurements (areas and volumes) of the lumen, total vessel, plaque-media complex, and percent obstruction were compared with findings by manual tracing of the IVUS images and of the corresponding histologic cross sections obtained at 2-mm increments (n = 100). Both area and volume measurements by the contour detection system agreed well with the results obtained by manual tracing, showing low mean between-method differences (-3.7% to 0.3%) with SDs not exceeding 6% and high correlation coefficients (r = 0.97 to 0.99). Measurements of the lumen, total vessel, plaque-media complex, and percent obstruction by the contour detection system correlated well with histomorphometry of areas (r = 0.94, 0.88, 0.80, and 0.88) and volumes (r = 0.98, 0.91, 0.83, and 0.91). Systematic differences between the results by the contour detection system and histomorphometry (29%, 13%, -9%, and -22%, respectively) were found, most likely resulting from shrinkage during tissue fixation. The results of this study indicate that this computerized IVUS analysis system is reliable for the assessment of coronary atherosclerosis in vivo.