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Nat Methods. 2019 Oct;16(10):1054-1062. doi: 10.1038/s41592-019-0579-4. Epub 2019 Sep 27.

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

Author information

1
Laboratory for Functional Optical Imaging, Department of Biomedical Engineering, Columbia University, New York, NY, USA.
2
Mortimer B. Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY, USA.
3
Department of Pediatrics, College of Physicians & Surgeons, Columbia University, New York, NY, USA.
4
Graduate School of Natural Sciences, Nagoya City University, Nagoya, Japan.
5
Center for Advanced Intelligence Project, RIKEN, Tokyo, Japan.
6
Laboratory for Functional Optical Imaging, Department of Biomedical Engineering, Columbia University, New York, NY, USA. Elizabeth.hillman@columbia.edu.
7
Mortimer B. Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY, USA. Elizabeth.hillman@columbia.edu.
8
Department of Radiology, Columbia University Medical Center and New York-Presbyterian Hospital New York, New York, NY, USA. Elizabeth.hillman@columbia.edu.

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.

PMID:
31562489
DOI:
10.1038/s41592-019-0579-4

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