The success of osseointegrated biomaterials often depends on the functional interface between the implant and mineralized bone tissue. Several parallels between natural and synthetic interfaces exist on various length scales from the microscale toward the cellular and the atomic scale structure. Interest lies in the development of more sophisticated methods to probe these hierarchical levels in tissues at both biomaterials interfaces and natural tissue interphases. This review will highlight new and emerging perspectives toward understanding mineralized tissues, particularly bone tissue, and interfaces between bone and engineered biomaterials at multilength scales and with multidimensionality. Emphasis will be placed on highlighting novel and correlative X-ray, ion, and electron beam imaging approaches, such as electron tomography, atom probe tomography, and in situ microscopies, as well as spectroscopic and mechanical characterizations. These less conventional approaches to imaging biomaterials are contributing to the evolution of the understanding of the structure and organization in bone and bone integrating materials.