This study aimed to determine the structural features between immature and mature articular cartilage from the humeral and femoral joints of rabbits. Specimens of articular cartilage (n = 6 for immature tissue, n = 6 for mature tissue) that were still attached to the underlying bone from a humerus (shoulder joint) or femur (knee joint) were imaged using microscopic MRI (µMRI) and polarized light microscopy (PLM). Quantitative µMRI data with a pixel resolution of 11.7-13.2 µm revealed a number of differences between the immature and mature cartilage, including total thickness, and T2 and T1ρ relaxation values. Quantitative PLM data with a pixel resolution of 0.25-1 µm confirmed the µMRI results and revealed additional differences in cellular features between the tissues. The mature cartilage had a clearly defined tidemark, which was absent in the immature tissue. The ability to differentiate specific maturation-related cartilage characteristics could be beneficial to translational studies of degenerative diseases such as osteoarthritis.