Using an on-the-fly scanning scheme, line confocal microscopy can obtain complex structures of large biological tissues with high throughput. Yet, it suffers from lateral imaging asymmetry and thus introduces the potential deformations of the observation results. Here, we propose cross-line illumination microscopy (cLIM) that acquires the imaging data of two perpendicular directions simultaneously through the same objective lens in a line scanning and utilizes two-direction deconvolution fusion to achieve lateral symmetric imaging performance. Imaging fluorescence beads indicates that cLIM reduces lateral resolution asymmetry from 46.1% to 2.5% and improves lateral resolution by 31.0%, compared with traditional line-scanning imaging. Compared with commercial point-confocal microscopy, the cLIM has a 25.84× increase in imaging speed and 1.93× better background-suppressing ability when imaging an 11,306 μm×7783 μm×100 μm mouse kidney slice. We also show the advantages of the cLIM in observing direction-sensitive texture features by imaging a muscular tissue slice. cLIM offers a novel solution to achieve laterally symmetric line-scanning imaging with simple modifications while maintaining high throughput and accuracy for imaging large-scale samples.

中文翻译：

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使用同步十字线照明进行横向对称成像的线扫描显微镜


使用动态扫描方案，线共焦显微镜可以高通量获得大型生物组织的复杂结构。然而，它受到横向成像不对称的影响，从而引入了观测结果的潜在变形。在这里，我们提出了十字线照明显微镜（cLIM），它通过同一物镜在线扫描同时获取两个垂直方向的成像数据，并利用两个方向反卷积融合来实现横向对称成像性能。成像荧光珠表明，与传统线扫描成像相比，cLIM 将横向分辨率不对称性从 46.1% 降低至 2.5%，横向分辨率提高 31.0%。与商用点共焦显微镜相比，cLIM 在对 11,306 像素进行成像时，成像速度提高了 25.84 倍，背景抑制能力提高了 1.93 倍。微米×7783微米×100微米小鼠肾切片。我们还展示了 cLIM 在通过对肌肉组织切片成像来观察方向敏感纹理特征方面的优势。 cLIM 提供了一种新颖的解决方案，通过简单的修改即可实现横向对称线扫描成像，同时保持大规模样品成像的高通量和准确性。