Multi-scale tissue fluorescence mapping with fiber optic ultraviolet excitation and generative modeling

Abstract

Cellular imaging of thick samples requires physical sectioning or laser scanning microscopy, which can be restrictive, involved, and generally incompatible with high-throughput requirements. We developed fiber optic microscopy with ultraviolet (UV) surface excitation (FUSE), a portable and quantitative fluorescence imaging platform for thick tissue that enabled quick sub-cellular imaging without thin sections. We substantially advanced prior UV excitation approaches with illumination engineering and computational methods. Optical fibers delivered <300nm light with directional control, enabling unprecedented 50× widefield imaging on thick tissue with sub-nuclear clarity, and 3D topography of surface microstructure. Probabilistic modeling of high-magnification images using our normalizing flow architecture FUSE-Flow (made freely available as open-source software) enhanced low-magnification imaging with measurable localized uncertainty via variational inference. Comprehensive validation comprised multi-scale fluorescence histology compared with standard H&E histology, and quantitative analyses of senescence, antibiotic toxicity, and nuclear DNA content in tissue models via efficient sampling of thick slices from entire murine organs up to 0.4×8×12mm and 1.3 million cells per surface. This technology addresses long-standing laboratory gaps in high-throughput studies for rapid cellular insights.