A Multi-Modal Light Sheet Microscope for High-Resolution 3D Tomographic Imaging with Enhanced Raman Scattering and Computational Denoising

Three-dimensional (3D) cellular models, such as spheroids, serve as pivotal systems for understanding complex biological phenomena in histology, oncology, and tissue engineering. In response to the growing need for advanced imaging capabilities, we present a novel multi-modal Raman light sheet microscope designed to capture elastic (Rayleigh) and inelastic (Raman) scattering, along with fluorescence signals, in a single platform. By leveraging a shorter excitation wavelength (532 nm) to boost Raman scattering efficiency and incorporating robust fluorescence suppression, the system achieves label-free, high-resolution tomographic imaging without the drawbacks commonly associated with near-infrared modalities. An accompanying Deep Image Prior (DIP) seamlessly integrates with the microscope to provide unsupervised denoising and resolution enhancement, preserving critical molecular details and minimizing extraneous artifacts. Altogether, this synergy of optical and computational strategies underscores the potential for in-depth, 3D imaging of biomolecular and structural features in complex specimens and sets the stage for future advancements in biomedical research, diagnostics, and therapeutics.