Hyperspectral Raman images of mineral components of trabecular and cortical bone at 3 mm spatial resolu- tion are presented. Contrast is generated from Raman spectra acquired over the 600–1400 cm−1 Raman shift range. Factor analysis on the ensemble of Raman spectra is used to generate descriptors of mineral compo- nents. In trabecular bone independent phosphate (PO₄−3) and monohydrogen phosphate (HPO₄−2) factors are observed. Phosphate and monohydrogen phosphate gradients extend from trabecular packets into the inte- rior of a rod. The gradients are sharply defined in newly regenerated bone. There, HPO₄−2 content maximizes near a trabecular packet and decreases to a minimum value over as little as a 20 mm distance. Incomplete mineralization is clearly visible. In cortical bone, factor analysis yields only a single mineral factor containing both PO₄−3 and HPO₄−2 signatures and this implies uniform distribution of these ions in the region imaged. Uniform PO₄−3 and HPO₄−2 distribution is verified by spectral band integration.
Keywords:
Raman imaging; cortical bone; trabecular bone; microscopy