The University of Oklahoma 1 article published in JoVE Bioengineering Layer Microdissection of Tricuspid Valve Leaflets for Biaxial Mechanical Characterization and Microstructural Quantification Katherine M. Casey1, Devin W. Laurence1, Mulan Tang1, Chung-Hao Lee1,2 1Biomechanics and Biomaterials Design Laboratory (BBDL), School of Aerospace and Mechanical Engineering, The University of Oklahoma, 2Institute for Biomedical Engineering, Science and Technology (IBEST), The University of Oklahoma This protocol describes the biaxial mechanical characterization, polarized spatial frequency domain imaging-based collagen quantification, and microdissection of tricuspid valve leaflets. The provided method elucidates how the leaflet layers contribute to the holistic leaflet behaviors.