Toward cancer theranostics via multimodal nonlinear endomicroscopy, microspectroscopy and femtosecond laser ablation

This thesis advances nonlinear multimodal imaging techniques, including Coherent Anti-Stokes Raman Scattering (CARS), Two-Photon Excited Fluorescence (TPEF), Second Harmonic Generation (SHG), and broadband Coherent Raman Scattering (BCARS), toward clinical applications to cancer diagnosis and treatment. A multimodal endomicroscopic platform integrating CARS, TPEF, SHG, indocyanine green (ICG) fluorescence, and femtosecond laser ablation was developed and validated for real-time cancer detection and treatment. The system enables a “seek-and-treat” approach combining imaging, AI-based tissue analysis, and selective laser ablation for image-guided surgery. Preclinical studies on head and neck cancer tissue demonstrated high-quality imaging and robust performance comparable to commercial systems. AI-based analysis achieved 88% sensitivity in distinguishing tissue requiring resection from healthy tissue, while automated image-guided laser ablation demonstrated precise selective tissue removal. The thesis also investigates broadband CARS for enhanced chemical specificity and spectral histopathology. BCARS systems were developed and applied to biological samples, showing strong potential for molecular characterization, tissue classification, and disease detection. Preliminary results further demonstrate the feasibility of integrating BCARS into endomicroscopy, supporting future real-time intraoperative spectral imaging. Overall, this work highlights the potential of advanced nonlinear imaging for image-guided cancer diagnosis and treatment.