Scanning transmission electron microscopic tomography of cortical bone using Z-contrast imaging

•HAADF STEM tomography is appropriate method for imaging bone ultrastructure.•Tomographic images of cortical bone confirm arrangement of mineral structures around fibrils and lack of abundant mineral inside fibrils.•Cross sectional and longitudinal (parallel to axis) images provide complementary but mutually consistent data.

Previously we presented (McNally et al., 2012) a model for the ultrastructure of bone showing that the mineral resides principally outside collagen fibrils in the form of 5 nm thick mineral structures hundreds of nanometers long oriented parallel to the fibrils. Here we use high-angle annular dark-field electron tomography in the scanning transmission electron microscope to confirm this model and further elucidate the composite structure. Views of a section cut parallel to the fibril axes show bundles of mineral structures extending parallel to the fibrils and encircling them. The mineral density inside the fibrils is too low to be visualized in these tomographic images. A section cut perpendicular to the fibril axes, shows quasi-circular walls composed of mineral structures, wrapping around apparently empty holes marking the sites of fibrils. These images confirm our original model that the majority of mineral in bone resides outside the collagen fibrils.