大视场高分辨率、大体积成像和多视场单物镜光片显微镜。

DaXi-high-resolution, large imaging volume and multi-view single-objective light-sheet microscopy.

机构信息

Chan Zuckerberg Biohub, San Francisco, CA, USA.

Calico Life Sciences LLC, South San Francisco, CA, USA.

出版信息

Nat Methods. 2022 Apr;19(4):461-469. doi: 10.1038/s41592-022-01417-2. Epub 2022 Mar 21.

Abstract

The promise of single-objective light-sheet microscopy is to combine the convenience of standard single-objective microscopes with the speed, coverage, resolution and gentleness of light-sheet microscopes. We present DaXi, a single-objective light-sheet microscope design based on oblique plane illumination that achieves: (1) a wider field of view and high-resolution imaging via a custom remote focusing objective; (2) fast volumetric imaging over larger volumes without compromising image quality or necessitating tiled acquisition; (3) fuller image coverage for large samples via multi-view imaging and (4) higher throughput multi-well imaging via remote coverslip placement. Our instrument achieves a resolution of 450 nm laterally and 2 μm axially over an imaging volume of 3,000 × 800 × 300 μm. We demonstrate the speed, field of view, resolution and versatility of our instrument by imaging various systems, including Drosophila egg chamber development, zebrafish whole-brain activity and zebrafish embryonic development - up to nine embryos at a time.

摘要

单目标光片显微镜的承诺是将标准单目标显微镜的便利性与光片显微镜的速度、覆盖范围、分辨率和温和性结合起来。我们提出了 DaXi,这是一种基于斜平面照明的单目标光片显微镜设计,实现了:(1)通过定制的远程聚焦物镜实现更宽的视野和高分辨率成像;(2)在不影响图像质量或不需要平铺采集的情况下,快速对更大体积进行体积成像;(3)通过多视角成像实现对大样本的更全面覆盖;(4)通过远程盖玻片放置实现更高的高通量多孔成像。我们的仪器在成像体积为 3000×800×300μm 时,实现了 450nm 横向和 2μm 轴向的分辨率。我们通过对各种系统进行成像,包括果蝇卵室发育、斑马鱼全脑活动和斑马鱼胚胎发育,证明了我们仪器的速度、视野、分辨率和多功能性——一次可同时对多达九个胚胎进行成像。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/975d/9007742/1719605839a9/41592_2022_1417_Fig1_HTML.jpg

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