Real-time volumetric microscopy of in vivo dynamics and large-scale samples with SCAPE 2.0
Real-time volumetric microscopy of in vivo dynamics and large-scale samples with SCAPE 2.0
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DOI：
10.1038/s41592-019-0579-4
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发表时间：
2019-10-01
期刊：
影响因子：
48
通讯作者：
Hillman, Elizabeth M. C.
中科院分区：
文献类型：
--
作者：
Voleti, Venkatakaushik;Patel, Kripa B.;Hillman, Elizabeth M. C.
大多数显微镜方法的有限的每像素带宽需要在视场、采样密度和成像速度之间进行折衷。这一限制限制了涉及复杂运动或快速细胞信号传导的研究，并成为高通量结构成像的主要瓶颈。在这里，我们结合联合收割机高速增强相机技术与一个多功能的，可重新配置和显着改善扫描，共焦对准平面激发（SCAPE）显微镜设计，可以实现高分辨率的体积成像超过300体积每秒和超过1.2 GHz的像素速率。我们展示了近各向同性采样自由移动的秀丽隐杆线虫，并分析实时血流和钙动力学跳动的斑马鱼心脏。同一系统还允许对封固的、完整的、清除的和扩增的样品进行高通量结构成像。SCAPE 2.0的显着较低的光损伤相比，点扫描技术也得到证实。我们的研究结果表明，SCAPE 2.0是一个强大的，但可访问的成像平台，无数新兴的高速动态和高通量体积显微镜应用。
The limited per-pixel bandwidth of most microscopy methods requires compromises between field of view, sampling density and imaging speed. This limitation constrains studies involving complex motion or fast cellular signaling, and presents a major bottleneck for high-throughput structural imaging. Here, we combine high-speed intensified camera technology with a versatile, reconfigurable and dramatically improved Swept, Confocally Aligned Planar Excitation (SCAPE) microscope design that can achieve high-resolution volumetric imaging at over 300 volumes per second and over 1.2 GHz pixel rates. We demonstrate near-isotropic sampling in freely moving Caenorhabditis elegans, and analyze real-time blood flow and calcium dynamics in the beating zebrafish heart. The same system also permits high-throughput structural imaging of mounted, intact, cleared and expanded samples. SCAPE 2.0's significantly lower photodamage compared to point-scanning techniques is also confirmed. Our results demonstrate that SCAPE 2.0 is a powerful, yet accessible imaging platform for myriad emerging high-speed dynamic and high-throughput volumetric microscopy applications.