Thinner and Longer Working Distance Light Sheet Illumination and Microscopic Imaging

Abstract

Light sheet fluorescence microscopy has become a basic tool in biology and medical research with its fast imaging speed, low phototoxicity and high spatiotemporal resolution. Here, we report a novel method to generate multiplane parallel light sheets with a micro structure named isosceles triangular array. The thickness of light sheet in each plane can approach 3.12 μm along with the working distance of 152.20 μm directly generated with the isosceles triangular array. We experimentally recorded the multiplane parallel light sheets illumination, which is consistent with the corresponding simulation by using ray-tracing method. To evaluate the imaging quality, we compared the light field generated with and without projected phase mask onto the spatial light modulator by using polystyrene fluorescent microspheres nanoparticles and transgenic zebrafish. This study provides a potential method for the building of volumetric imaging light sheet fluorescence microscopy with high resolution and low phototoxicity.