High-frequency ultrasound can potentially display gross morphologic changes during thrombus formation and lysis. Current intravascular ultrasound (IVUS) devices, however, provide only 2-dimensional cross-sectional images with limited overall appreciation of thrombus size and 3-dimensional (3D) configuration. The purpose of this study was to explore the ability of 3D reconstruction of serial ultrasound images to provide a quantitative assessment of intravascular thrombi. We therefore imaged 11 arterial thrombi of varying shape and volume (10 to 116mm3). To avoid thrombus disruption, we used an epivascular approach (also suitable for transvenous imaging) with a 20MHz IVUS catheter withdrawn at 1mm/sec. A 3D voxel image intensity data set was reconstructed, and thrombus volume was semiautomatically extracted based on its intensity. Calculated volume was compared with directly measured values by volume displacement in a miniature cylinder.3D reconstruction provided previously unobtainable longitudinal and 3D views that improved spatial appreciation of thrombus size, shape and channel formation. Calculated thrombus volumes agreed well with actual volumes: y=0.92x+2.4, r=0.98, SEE=5mm3, mean error = 1±5mm3(ns vs 0).3D reconstruction can improve spatial appreciation of the shape of thrombi and accurately measure their volumes. This approach, suitable for epivascular or transvenous imaging, could potentially be used to study thrombus formation and lysis in research and clinical studies.