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.

Voleti, Venkatakaushik;Patel, Kripa B;Li, Wenze;Perez Campos, Citlali;Bharadwaj, Srinidhi;Yu, Hang;Ford, Caitlin;Casper, Malte J;Yan, Richard Wenwei;Liang, Wenxuan;Wen, Chentao;Kimura, Koutarou D;Targoff, Kimara L;Hillman, Elizabeth M C;
Nature Methods 2019 Vol. 16 pp. 1054-1062
355
voleti2019realtimenature

Abstract

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.

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