OBJECTIVE. Our goal was to determine how the three-dimensional structure of hyaline cartilage affects its MR appearance and to correlate this appearance with detailed structural analysis using scanning electron microscopy and freeze-fracture sectioning techniques. MATERIALS AND METHODS. In vitro 7-T spin-echo MR images of hyaline cartilage specimens from four patients undergoing above-knee amputations were obtained parallel and perpendicular to the main magnetic field. Specimens were imaged with low- and high-power scanning electron microscopy after freeze fracturing. The corresponding images from both techniques were analyzed with specific attention to the three-dimensional structure of the cartilage, collagen fibril orientation, and respective changes in the MR appearance. RESULTS. Freeze fracturing of cartilage reveals a curved fracture plane. Expected changes in signal intensity predicted by the magic angle effect correlated with observed changes in signal intensity across the thickness of the sample. Changes in individual collagen fibril orientation did not correspond to MR layering. CONCLUSION. The three-dimensional organization of collagen in cartilage has a strong influence on the MR appearance of cartilage. This influence is caused by the restriction of water mobility and the resulting magic angle effect caused by curvature of the collagen network, possibly because of the influence on proteoglycan orientation.