The goal of this study was to demonstrate the feasibility of using 7-Tesla (7T) magnetic resonance imaging (MRI) and micro-finite element analysis (µFEA) to evaluate mechanical and structural properties of whole, cortical, and trabecular bone at the distal femur and proximal tibia in vivo. 14 healthy subjects were recruited (age 40.7 ± 15.7 years). The right knee was scanned on a 7T MRI scanner using a 28 channel-receive knee coil and a three-dimensional fast low-angle shot sequence (TR/TE 20 ms/5.02 ms, 0.234 mm × 0.234 mm × 1 mm, 80 axial images, 7 min 9 s). Bone was analyzed at the distal femoral metaphysis, femoral condyles, and tibial plateau. Whole, cortical, and trabecular bone stiffness was computed using µFEA. Bone volume fraction (BVF), bone areas, and cortical thickness were measured. Trabecular bone stiffness (933.7 ± 433.3 MPa) was greater than cortical bone stiffness (216 ± 152 MPa) at all three locations (P < 0.05). Across locations, there were no differences in bone stiffness (whole, cortical, or trabecular). Whole, cortical, and trabecular bone stiffness correlated with BVF (R ≥ 0.69, P < 0.05) and inversely correlated with corresponding whole, cortical, and trabecular areas (R ≤ -0.54, P < 0.05), but not with cortical thickness (R < -0.11, P > 0.05). Whole, cortical, and trabecular stiffness correlated with body mass index (R ≥ 0.62, P < 0.05). In conclusion, at the distal femur and proximal tibia, trabecular bone contributes 66-74% of whole bone stiffness. 7T MRI and µFEA may be used as a method to provide insight into how structural properties of cortical or trabecular bone affect bone mechanical competence in vivo.