Quantitative changes in human epithelial cancers and osteogenesis imperfecta disease detected using nonlinear multicontrast microscopy

Abstract

Abstract. We show that combined multimodal nonlinear optical (NLO) microscopies, including two-photon excitation fluorescence, second-harmonic generation (SHG), third harmonic generation, and fluorescence lifetime imaging microscopy (FLIM) can be used to detect morphological and metabolic changes associated with stroma and epithelial transformation during the progression of cancer and osteogenesis imperfecta (OI) disease. NLO microscopes provide complementary information about tissue microstructure, showing distinctive patterns for different types of human breast cancer, mucinous ovarian tumors, and skin dermis of patients with OI. Using a set of scoring methods (anisotropy, correlation, uniformity, entropy, and lifetime components), we found significant differences in the content, distribution and organization of collagen fibrils in the stroma of breast and ovary as well as in the dermis of skin. We suggest that our results provide a framework for using NLO techniques as a clinical diagnostic tool for human cancer and OI. We further suggest that the SHG and FLIM metrics described could be applied to other connective or epithelial tissue disorders that are characterized by abnormal cells proliferation and collagen assembly.

Differentiating the two main histologic categories of fibroadenoma tissue from normal breast tissue by using multiphoton microscopy

“…TPEF is resulting from the nonlinear excitation of molecular fluorescence and that can visualize tissue architecture and cellular morphology. In addition to TPEF, SHG is well suited to observe collagen fibre structure in extracellular matrices that allows MPM to monitor changes in collagen composition during tumour development, whereas the changes were not obvious by standard histopathology (Adur et al ., ; Chen et al ., ). Due to the autofluorescence of properties in breast tissue, recent studies showed that the miniaturized MPM and multiphoton probe have potential ability for the noninvasive in vivo pathological diagnostic of breast tumour (Wu et al ., ).…”

The Severity of Osteogenesis Imperfecta and Type I Collagen Pattern in Human Skin as Determined by Nonlinear Microscopy: Proof of Principle of a Diagnostic Method

“…Both techniques have already been widely used on ex-vivo tissue samples to perform optical biopsy for morphological characterizations [12], [13], [21]. However, there are only a few reports in the literature using SHG [22], [23] or TPEF+SHG [24] to analyze OI in skin samples.…”

Nonlinear Microscopy Techniques: Principles and Biomedical Applications

“…In contrast to the chemical specificity that characterizes fluorescence images, harmonic generation (SHG and THG) provides an imaging modality specific for structural configuration. In the study of cancer tumors, our experience with both techniques is that SHG is an excellent tool to observe collagen network of extracellular matrix, while THG allows to clearly display the nuclei, which are two key parameters for pathologists [4,[18][19][20]. In addition to harmonic generation microscopy, CARS microscopy is another 3D highresolution imaging approach that circumvents exogenous probes.…”