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62 In sum, this analysis cannot resolve the small fibrils, but the coherence of the SHG permits inferences of organization on this sub-resolution size scale. SHG directionality measurements have also been extended to breast PRDX4 cancer, but the analyses have been inconclusive.63 This may be because the measurements were taken on thin histological slides and are susceptible to reflections and are also highly sensitive to any slight changes in thickness. In contrast, our combined imaging and simulation method requires tissues of 50�C100 ��m in thickness to properly account for scattering and are less sensitive to experimental artifacts. In addition, breast tissues are more heterogeneous than the ovarian stroma, which is comprised predominantly of dense collagen. Therefore, additional studies are required to obtain robust knowledge of the SHG directionality alterations associated with normal and malignant breast stroma. Figure 4 Depth-dependent SHG F/B measurements of normal and high-grade serous ovarian cancer. Best fits using Monte Carlo simulations and independently measured ��s and g resulted in 93% and 77% forward-directed SHG in normal and cancer respectively. Adapted ... The combined imaging/optical scattering/simulation approach requires several measurements and is computationally intensive; but it is highly general and may be effective in complex fibrillar organization when simpler morphology-based analyses (eg, transforms described earlier) are not applicable. To date, we have found collagen-based tissues have negligible absorption (secondary Duvelisib molecular weight filter), but we anticipate blood-rich organs will have significant absorption due to the overlap of SHG photons and the absorption spectrum of blood. In performing the SHG emission directionality studies of blood-rich organs, we would account for the increased absorption (measured by integrating spheres) in our Monte Carlo simulations to accurately model the response. An analogous response has long been done in diffuse optical tomography in reconstructing the image.64 For example, we have used this approach for the analysis of the connective tissue disorder osteogenesis imperfecta60 and also in the analysis of optical clearing.61 To date, TPEF and SHG imaging studies have provided a wealth of information concerning the Bleomycin cell line early ECM alterations associated with breast and ovarian cancers. Tables 2 and ?and33 summarize the results and conclusions of several papers. Overall, these studies indicate that TPEF and SHG imaging has the sensitivity and specificity to detect early cancerous lesions for both cancers. Table 2 Summary of TPEF and SHG imaging studies and techniques to probe collagen remodeling of the ECM in ovarian cancer. Table 3 Summary of TPEF and SHG imaging studies and techniques to probe collagen remodeling of the ECM in breast cancer.