Two-photon light-sheet microscopy offers high resolution and low photobleaching,making it widely used in biological research.However,a trade-off between light sheet thickness and length limits its axial resolution and field of view,restricting its applications.To address this issue,this paper employs a liquid crystal spatial light modulator(SLM)to modulate conventional Gaussian illumination beam into Bessel beam.The"non-diffracting"property of the Bessel beam enables the generation of a large and thin light sheet,achieving a length-to-thickness ratio of 163.Based on this,a Bessel two-photon light-sheet microscope is constructed,achieving three-dimensional high-resolution imaging with a lateral resolution of 440 nm and an axial resolution of 1.9 μm within a 310 μm field of view.Successful in vivo imaging of zebrafish embryos is demonstrated,including the measurement of blood cell flow velocity and the segmentation and counting of spinal cord cells,validating the application potential of the Bessel two-photon light-sheet microscope in live biological imaging.
维普
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