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5 August 2019 Propagating-path uniformly scanned light sheet excitation microscopy for isotropic volumetric imaging of large specimens
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Abstract

We demonstrate a propagating-path uniformly scanned light sheet excitation (PULSE) microscopy based on the oscillation of voice coil motor that can rapidly drive a thin light sheet along its propagation direction. By synchronizing the rolling shutter of a camera with the motion of laser sheet, we can obtain a uniform plane-illuminated image far beyond the confocal range of Gaussian beam. A stable 1.7-μm optical sectioning under a 3.3  mm  ×  3.3  mm wide field of view (FOV) has been achieved for up to 20 Hz volumetric imaging of large biological specimens. PULSE method transforms the extent of plane illumination from one intrinsically limited by the short confocal range (μm scale) to one defined by the motor oscillation range (mm scale). Compared to the conventional Gaussian light sheet imaging, our method greatly mitigates the compromise of axial resolution and successfully extends the FOV over 100 times. We demonstrate the applications of PULSE method by rapidly imaging cleared mouse spinal cord and live zebrafish larva at isotropic subcellular resolution.

CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
Junyu Ping, Fang Zhao, Jun Nie, Tingting Yu, Dan Zhu, Mugen Liu, and Peng Fei "Propagating-path uniformly scanned light sheet excitation microscopy for isotropic volumetric imaging of large specimens," Journal of Biomedical Optics 24(8), 086501 (5 August 2019). https://doi.org/10.1117/1.JBO.24.8.086501
Received: 4 June 2019; Accepted: 15 July 2019; Published: 5 August 2019
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CITATIONS
Cited by 8 scholarly publications and 2 patents.
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KEYWORDS
Confocal microscopy

Microscopy

Image resolution

Cameras

Biomedical optics

Camera shutters

Luminescence

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