BACKGROUND We aimed to develop and evaluate a deep learning-based method for fully automatic segmentation of knee joint MR imaging and quantitative computation of knee osteoarthritis (OA)-related imaging biomarkers. MATERIAL AND METHODS This retrospective study included 843 volumes of proton density-weighted fat suppression MR imaging. A convolutional neural network segmentation method with multiclass gradient harmonized Dice loss was trained and evaluated on 500 and 137 volumes, respectively. To assess potential morphologic biomarkers for OA, the volumes and thickness of cartilage and meniscus, and minimal joint space width (mJSW) were automatically computed and compared between 128 OA and 162 control data. RESULTS The CNN segmentation model produced reasonably high Dice coefficients, ranging from 0.948 to 0.974 for knee bone compartments, 0.717 to 0.809 for cartilage, and 0.846 for both lateral and medial menisci. The OA-related biomarkers computed from automatic knee segmentation achieved strong correlation with those from manual segmentation: average intraclass correlations of 0.916, 0.899, and 0.876 for volume and thickness of cartilage, meniscus, and mJSW, respectively. Volume and thickness measurements of cartilage and mJSW were strongly correlated with knee OA progression. CONCLUSIONS We present a fully automatic CNN-based knee segmentation system for fast and accurate evaluation of knee joint images, and OA-related biomarkers such as cartilage thickness and mJSW were reliably computed and visualized in 3D. The results show that the CNN model can serve as an assistant tool for radiologists and orthopedic surgeons in clinical practice and basic research.