In this study nonlinear microscopy techniques utilized to shed new light in breast cancer diagnosis. In particular, the nonlinear imaging modalities of multi-photon excitation fluorescence (MPEF) second and third harmonic generation (SHG, THG) used as non-destructive, label free diagnostic tools for the qualitative discrimination of breast cancer tissues. Nonlinear signals were collected from unstained thin histological sections of breast biopsies samples. In an attempt to discriminate the different types of tissues, quantitative analysis of the recorded THG signals on specific cells in tissue was also performed.
Non-linear optical imaging techniques have been used to greatly enhance our understanding of issues with high biological significance and promise a strong impact on early and accurate detection of various diseases. In our current work, we employ Third Harmonic Generation (THG) and Second Harmonic Generation (SHG) imaging modalities for diagnosis of cancerous tissue limits and for obtaining additional quantitative information to supplement standard histopathology procedures. For this reason, unstained histological slides of breast tumor biopsies were irradiated. Cancerous and normal tissue areas could be distinguished based on cell morphology, size, and density. THG imaging reflects lipid bodies (LBs) in the intracellular compartments, cellular and nuclear membranes, while SHG shows collagen distribution in the tissue. By using THG microscopy, it is feasible to concentrate on specific cells in tissue and collect quantitative information. Our initial results showed that quantification of THG signaling can depict differences between healthy and cancerous tissue. This is a very promising observation, since the non-linear technique described here, allows fast, non-invasive, label-free imaging that does not require the use of fluorescent dyes or other preparations of tissues in order to detect specific structures and features. The significance of this work has a clinical potential since it can monitor quantitative changes in cellular behavior in healthy and pathological human tissues.
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