  Currently, a large number of stored tissue samples are unavailable for
spectroscopic study without the time consuming and destructive process of
paraffin removal. Instead, a structurally sensitive technique, sum frequency
generation, and a chemically sensitive technique, coherent anti-Stokes Raman
scattering enables imaging through the paraffin. This method is demonstrated by
imaging collagen in mouse tibia. We introduce a statistical method for
separating images by quality and, with the aid of machine learning, distinguish
osteoporotic and healthy bone. This method has the potential to verify the
results of previous studies and reduce new sample production by allowing
retesting results with spectroscopy.
