The objective of this study was to create three-dimensional (3D) images for the histomorphological study of osteons. Medical imaging technology was used to register digitized 2D images of serial decalcified histological sections of bone, to segment the tissues of interest from the surrounding tissues, and to create 3D reconstructions from the segmented structures. Examination of the 3D reconstructions did not support suggestions in the literature that osteons have a spiraling organization. In contrast, the 3D reconstructions indicated that osteons have a complex pattern of organization that is dominated by branching. Examination of the reconstructions also suggested that osteons described in the literature as being dumbbell shaped are actually artifacts of the plane of sectioning. This study demonstrated the applicability of imaging and visualization technology developed for the 3D reconstruction of medical images to the reconstruction of digitized 2D images of serial sections Of bone and additionally demonstrated the feasibility of using 3D reconstructions for the histomorphological study of osteons.