Enhancing Total Optical Throughput of Microscopy with Deep Learning for Intravital Observation

Abstract The significance of performing large‐depth dynamic microscopic imaging in vivo for life science research cannot be overstated. However, the optical throughput of the microscope limits the available information per unit of time, i.e., it is difficult to obtain both high spatial and temporal resolution at once. Here, a method is proposed to construct a kind of intravital microscopy with high optical throughput, by making near‐infrared‐II (NIR‐II, 900–1880 nm) wide‐field fluorescence microscopy learn from two‐photon fluorescence microscopy based on a scale‐recurrent network. Using this upgraded NIR‐II fluorescence microscope, vessels in the opaque brain of a rodent are reconstructed three‐dimensionally. Five‐fold axial and thirteen‐fold lateral resolution improvements are achieved without sacrificing temporal resolution and light utilization. Also, tiny cerebral vessel dilatations in early acute respiratory failure mice are observed, with this high optical throughput NIR‐II microscope at an imaging speed of 30 fps.