We propose a method called spatial autocorrelation analysis (SACA) to determine the spatial anisotropy of the trabecular bone in order to investigate osteoporosis. For demonstrating the potential of SACA we first evaluate the method on rectangular, simulated test patterns as a simple model for the anisotropic pore structure of the bone. As a next step towards biomedical application, photographic reference images of human vertebral bone were investigated by SACA. Osteoporotic bone structure could be clearly differentiated from non-osteoporotic sample images. Moreover, for demonstration of the applicability and potential of the method for in vivo characterization of osteoporosis, the microstructure of the human calcaneus was investigated by MR-microimaging on a young healthy male subject and an osteoporotic female. The measurements were performed using a high-field (3T) whole-body MR tomograph equipped with a special, strong head gradient system. The signal was acquired with a surface coil mounted on an in-house-built device for convenient immobilization of the subject's foot. Using a 3D gradient echo sequence a resolution of 0.254 × 0.254 × 2.188 mm3 was achieved in vivo. Selected images were inverted, gradient corrected for the inhomogeneous but sensitive detection by the surface coil, and subsequently analyzed by SACA. The anisotropy of bone structure detected by SACA is a possible candidate for noninvasive determination of the osteoporotic status, potentially complementing standard bone mineral density measurements. J. Magn. Reson. Imaging 2001;14:87–93. © 2001 Wiley-Liss, Inc.