Multiple airy beams light-sheet fluorescence microscopy
Multiple airy beams light-sheet fluorescence microscopy
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DOI：
10.3389/fphy.2022.1111023
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发表时间：
2022-12
期刊：
影响因子：
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通讯作者：
Shuangyu Gu;Xianghua Yu;Chen Bai;Junwei Min;Runze Li;Yanlong Yang;Baoli Yao
中科院分区：
文献类型：
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作者：
Shuangyu Gu;Xianghua Yu;Chen Bai;Junwei Min;Runze Li;Yanlong Yang;Baoli Yao
光片荧光显微镜(LSFM)是一种适用于长时间、高时空分辨率的活体标本的体积成像方法。单艾里光束(SAB)光片可以利用其无衍射的性质扩展光片荧光显微镜的视场，但代价是旁瓣照明导致背景散焦和成像对比度低。在这里，我们提出了一种由多个不同尺度因子的单艾利束线性叠加而成的多艾利束的产生方法。与SAB光片相比，多束艾里光光片的能量更集中在探测物镜的焦面上，可以提高成像对比度，减小光损伤效应。此外，我们结合互补波束减法来提高轴向分辨率，称为多艾里波束-互补波束减法，该方法在保持450μm×450μm视场的同时，使轴向分辨率达到1.2μm。
Light-sheet fluorescence microscopy (LSFM) is a kind of volumetric imaging methodology suited for long term living specimens at high temporal-spatial resolution. A single Airy beam (SAB) light-sheet can extend the field of view of Light-sheet fluorescence microscopy benefiting from its non-diffracting nature, but at the cost of out-of-focus background and low imaging contrast caused by side lobes illumination. Here, we propose a method to generate a sort of multiple Airy beams (MAB), which are linearly superimposed of multiple single Airy beams with different scale factors. Compared to the SAB light-sheet, the energy of the multiple Airy beams light-sheet is more concentrated on the focal plane of the detection objective, which can improve the imaging contrast and decrease the photodamage effect. Furthermore, we combined the complementary beam subtraction (CBS) strategy to increase the axial resolution, termed as multiple Airy beams-complementary beam subtraction method, which enables the axial resolution of 1.2 μm while keeping the field of view of 450 μm × 450 μm. The effectiveness of the method is demonstrated by imaging of fluorescent beads and aspergillus conidiophores.