The mechanical properties of soft tissues are driven by their complex, heterogeneous composition and structure. Interestingly, studies of soft tissue biomechanics often ignore spatial heterogeneity. In our work, we are therefore interested in exploring the impact of tissue heterogeneity on the mechanical properties of soft tissues. Therein, we specifically focus on soft tissue heterogeneity arising from spatially-varying thickness. To this end, our first goal is to develop a non-destructive measurement technique that has a high spatial resolution, provides continuous thickness maps, and is fast. Our secondary goal is to demonstrate that including spatial variation in thickness is important to the accuracy of biomechanical analyses. To this end, we use mitral valve leaflet tissue as our model system. To attain our first goal, we identify a soft tissue-specific contrast protocol that enables thickness measurements using a Keyence profilometer. We also show that this protocol does not affect our tissues' mechanical properties. To attain our second goal, we conduct virtual biaxial, bending, and buckling tests on our model tissue both ignoring and considering spatial variation in thickness. Thereby, we show that the assumption of average, homogeneous thickness distributions significantly alters the results of biomechanical analyses when compared to including true, spatially-varying thickness distributions. In conclusion, our work provides a novel measurement technique that can capture continuous thickness maps non-invasively, at high resolution, and in short time. Our work also demonstrates the importance of including heterogeneous thickness in biomechanical analyses of soft tissues.