Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy
Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy
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DOI：
10.1038/nmeth.2062
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发表时间：
2012-07-01
期刊：
影响因子：
48
通讯作者：
Keller, Philipp J.
中科院分区：
文献类型：
--
作者：
Tomer, Raju;Khairy, Khaled;Keller, Philipp J.
大型生物样本的实时成像从根本上受到光学显微镜光学穿透深度短的限制。为了最大限度地扩大物理覆盖范围，我们开发了用于高速体内成像的 SiMView 技术框架，可同时记录标本的多个视图。 SiMView 由具有四个同步光学臂的光片显微镜、用于以每秒 1.75 亿体素的速度长期采集基于 sCMOS 的图像的实时电子设备以及用于高通量图像配准、分割、跟踪和实时管理每个样本记录的 TB 多视图数据的计算模块组成。我们开发了单光子和多光子 SiMView 实现，并在整个果蝇胚胎的整个发育过程中以 30 秒的时间分辨率记录了细胞动力学。我们还对发育中的神经系统进行了高分辨率长期成像，并跟踪了体内神经母细胞谱系。 SiMView 数据集甚至可以为快速全局过程提供定量形态信息，并能够在整个早期胚胎中进行准确的自动细胞跟踪。
Live imaging of large biological specimens is fundamentally limited by the short optical penetration depth of light microscopes. To maximize physical coverage, we developed the SiMView technology framework for high-speed in vivo imaging, which records multiple views of the specimen simultaneously. SiMView consists of a light-sheet microscope with four synchronized optical arms, real-time electronics for long-term sCMOS-based image acquisition at 175 million voxels per second, and computational modules for high-throughput image registration, segmentation, tracking and real-time management of the terabytes of multiview data recorded per specimen. We developed one-photon and multiphoton SiMView implementations and recorded cellular dynamics in entire Drosophila melanogaster embryos with 30-s temporal resolution throughout development. We furthermore performed high-resolution long-term imaging of the developing nervous system and followed neuroblast cell lineages in vivo. SiMView data sets provide quantitative morphological information even for fast global processes and enable accurate automated cell tracking in the entire early embryo.