Deep ultraviolet-excited fluorescence histology with fibre optic illumination for tissue microtopography and quantitative mapping
bioRxiv (Cold Spring Harbor Laboratory)(2022)
摘要
Cellular imaging of thick samples requires physical sectioning or laser scanning microscopy, generally incompatible with high-throughput requirements. We developed fibre optic microscopy with ultraviolet (UV) surface excitation (FUSE), a portable, quantitative fluorescence imaging platform for thick tissue that substantially advances prior UV excitation approaches with illumination engineering and computational methods. Optic fibres delivered <300nm light with directional control, enabling unprecedented 50X widefield imaging on thick tissue with sub-nuclear clarity, and 3D topography of surface microstructure. Generative modelling of high-magnification images using our normalising flow architecture FUSE-Flow (open-source) enhanced low-magnification imaging by variational inference. Comprehensive validation comprised multi-scale fluorescence histology compared with standard H&E, and quantitative analyses of senescence, antibiotic toxicity, and nuclear DNA content in tissue models via efficient sampling of entire murine organs by thick slices up to 0.4x8x12mm and 1.3 million cells per surface. This technology addresses long-standing laboratory gaps for high-throughput studies for rapid cellular insights.
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关键词
fluorescence histology,tissue microtopography,ultraviolet-excited
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