Polarization-sensitive optical coherence tomography (PS-OCT) was applied to calcified aortic valves as well as bovine and porcine pericardium subjected to glutaraldehyde fixation and in-vitro calcification. Retardation, optic axis orientation and depolarization images revealed collagen layers, elastin fibers and micro-calcifications in 3D. PS-OCT enables non-destructive monitoring of structural and pathological changes in valve biomaterials.