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Multiscale SHG microscopy analysis of collagen alterations in human diseases

We have developed Second Harmonic Generation (SHG) microscope tools to selectively and specifically probe changes in collagen organization in diseased tissues. Using a novel form of 3D machine learning, we successfully classified six types of ovarian tumors based on the observed collagen fiber morphology. We also exploit the SHG coherence to extract sub-resolution fibril assembly (size and packing) by analyzing the emission directionality in conjunction with measured optical scattering properties and Monte Carlo simulations. This approach further classified a spectrum of ovarian tumors. We developed polarization sensitive SHG methods to extract collagen macro/supramolecular structural aspects and found significant differences between normal and malignant ovarian tissues. We also used this set of SHG analyses to probe structural changes in idiopathic pulmonary fibrosis (IPF) compared to normal lung tissues, and found comparable collagen changes over normal tissues.