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Real-time volumetric microscopy of in vivo dynamics and large-scale samples with SCAPE 2.0

2019; Nature Portfolio; Volume: 16; Issue: 10 Linguagem: Inglês

10.1038/s41592-019-0579-4

1548-7105

Venkatakaushik Voleti, Kripa Patel, Wenze Li, Citlali Pérez Campos, Srinidhi Bharadwaj, Hang Yu, Caitlin Ford, Malte Casper, Richard Wenwei Yan, Wenxuan Liang, Chentao Wen, Koutarou D. Kimura, Kimara L. Targoff, Elizabeth M. C. Hillman,

Single-cell and spatial transcriptomics

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. SCAPE 2.0 is a versatile imaging platform that enables real-time three-dimensional microscopy of cellular function and dynamic motion in living organisms at over 100 volumes per second with minimal photodamage, and high-throughput structural imaging in fixed, cleared and expanded samples.