The present invention relates to scanning confocal microscopy used to systematically quantify characteristic collagen fibril orientations by position within the lamellar thickness of secondary osteons along the osteon radial direction. Fully calcified lamellar specimens appear either extinct or bright in cross-section under circularly polarized light, and can be isolated from embedded osteon, flattened, and examined along the radial thickness direction of the original embedded osteon. Collagen orientation is measured from confocal image stacks. Extinct and bright lamellae display distinct patterns of collagen orientation distribution. Relative counts of collagen fibrils that are longitudinal to the osteon axis in extinct lamellae, transverse to the osteon axis in bright lamellae, and oblique to the osteon axis in both lamellar types, show parabolic distribution through the osteon radial direction.