Deep ultraviolet-excited fluorescence histology with fibre optic illumination for tissue microtopography and quantitative mapping

Abstract

1AbstractRapidly resolving microstructure and function from thick biological samples is a critical requirement from basic research to clinical diagnosis. The need for physical sectioning leads to limitations in the speed, cost, or ease of sample preparation and imaging. We introduce a fluorescence microscope with deep ultraviolet illumination via omnidirectional fibre optics, based on the concept of microscopy with ultraviolet surface excitation (MUSE). We further exploited precise control of directional illumination to produce 3D reconstructions of surface microtopography from 2D images. Our technology is applicable to diverse sample types, ranging from single cells and 3D cultures to large tissue samples while utilising conventional, widely available objective lenses in a compact and portable implementation (30cmby 30cm). We demonstrated the use of our microscope with important fluorescent assays through quantitative viability studies in single cells to whole organs, organ-level studies comparable to standard H&E histology, and multiplexed immunohistochemistry staining. We also explored the potential for microtopography to provide additional insight into vascular and tissue texture and heterogeneity studies. This technology fills long-standing technological gaps in both the laboratory and clinic for a general-purpose microscope that is simple, portable, and cheap that can provide near real-time biological insights.