This article first provides a brief review of the status of the subfield of three-dimensional (3D) materials analyses that combine serial sectioning, electron backscatter diffraction (EBSD), and finite element modeling (FEM) of materials microstructures, with emphasis on initial investigations and how they led to the current state of this research area. The discussions focus on studies of the mechanical properties of polycrystalline materials where 3D reconstructions of the microstructure—including crystallographic orientation information—are used as input into image-based 3D FEM simulations. The authors' recent work on a β-stabilized Ti alloy is utilized for specific examples to illustrate the capabilities of these experimental and modeling techniques, the challenges and the solutions associated with these methods, and the types of results and analyses that can be obtained by the close integration of experiments and simulations.