Chemical structure-based three-dimensional reconstruction of human cortical bone from two-dimensional infrared images

A method is described for the construction of three-dimensional infrared images from serial two-dimensional infrared images of various chemical structure parameters acquired from cortical bone. A series of serial cortical bone sections, each 4-5 mum thick, separated by similar to100 mum, were microtomed from PMMA-embedded normal human tibia. Twenty-three IR image cubes were acquired from this series, corresponding to an overall sample length of similar to2.2 mm. From each image cube, IR images of the spatial distribution of the mineral-to-matrix (protein) ratio, the mineral crystallinity, and the ratio of nonreducible-to-reducible collagen crosslinks were calculated. Two-dimensional images were stacked to form a 3D view of the cortical bone, with contrast provided by variation in the spatial distribution of any of the aforementioned chemical/molecular structural parameters.