Intravascular ultrasonography is emerging as an important imaging modality to assess the presence, distribution, and extent of atherosclerotic vascular disease. To determine the accuracy and clinical utility of intravascular ultrasonography, a flexible catheter-based system was used to generate two-dimensional, cross-sectional vascular images. In 23 arteries of 11 sheep 206 in vivo images demonstrated an echo-free lumen surrounded by three distinct concentric acoustic transitions corresponding to intima, media, and adventitia. Ultrasound measurements of lumen diameter and area correlated significantly with those of corresponding arteriographic measurements obtained by use of digital calipers (r = 0.91, r = 0.86). To evaluate clinical feasibility, intraoperative images (n = 160) were obtained in 10 patients undergoing vascular bypass or hemodialysis access procedures. The images depicted luminal configuration and arterial wall morphologic characteristics. Measurements of lumen diameter and lumen area correlated closely with corresponding intraoperative arteriography (r = 0.81, r = 0.79). The ultrasound images demonstrated arterial stenoses, intimal hyperplasia, intraluminal thrombus, polytetrafluoroethylene graft material, and anastomotic sites. We conclude that flexible catheter-based ultrasonography produces images that accurately demonstrate arterial wall architecture, lumen diameter, and area. Intraoperative application can produce images that provide unique information thus expanding the clinical potential of ultrasonography as a guidance system for vascular procedures.