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High-Resolution, Large Field-of-View, and Multi-View Single Objective Light-Sheet Microscopy

View ORCID ProfileBin Yang, Alfred Millett-Sikking, View ORCID ProfileMerlin Lange, Ahmet Can Solak, Hirofumi Kobayashi, Andrew York, View ORCID ProfileLoic A. Royer
doi: https://doi.org/10.1101/2020.09.22.309229
Bin Yang
1Chan Zuckerberg Biohub, San Francisco, USA
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  • For correspondence: loic.royer@czbiohub.org bin.yang@czbiohub.org
Alfred Millett-Sikking
2Calico Life Sciences LLC, South San Francisco, USA
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Merlin Lange
1Chan Zuckerberg Biohub, San Francisco, USA
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Ahmet Can Solak
1Chan Zuckerberg Biohub, San Francisco, USA
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Hirofumi Kobayashi
1Chan Zuckerberg Biohub, San Francisco, USA
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Andrew York
2Calico Life Sciences LLC, South San Francisco, USA
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Loic A. Royer
1Chan Zuckerberg Biohub, San Francisco, USA
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  • For correspondence: loic.royer@czbiohub.org bin.yang@czbiohub.org
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Abstract

Light-sheet microscopy has become the preferred method for long-term imaging of large living samples because of its low photo-invasiveness and good optical sectioning capabilities. Unfortunately, refraction and scattering often pose obstacles to light-sheet propagation and limit imaging depth. This is typically addressed by imaging multiple complementary views to obtain high and uniform image quality throughout the sample. However, multi-view imaging often requires complex multi-objective configurations that complicate sample mounting, or sample rotation that decreases imaging speed. Recent developments in single-objective light-sheet microscopy have shown that it is possible to achieve high spatio-temporal resolution with a single objective for both illumination and detection. Here we describe a single-objective light-sheet microscope that achieves: (i) high-resolution and large field-of-view imaging via a custom remote focusing objective; (ii) simpler design and ergonomics by remote placement of coverslips; (iii) fast volumetric imaging by means of light-sheet stabilised stage scanning – a novel scanning modality that extends the imaging volume without compromising imaging speed nor quality; (iv) multi-view imaging by means of dual orthogonal light-sheet illumination. Finally, we demonstrate the speed, field of view and resolution of our novel instrument by imaging zebrafish tail development.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • https://github.com/royerlab/hr-lf-mv-SOLS

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted September 24, 2020.
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High-Resolution, Large Field-of-View, and Multi-View Single Objective Light-Sheet Microscopy
Bin Yang, Alfred Millett-Sikking, Merlin Lange, Ahmet Can Solak, Hirofumi Kobayashi, Andrew York, Loic A. Royer
bioRxiv 2020.09.22.309229; doi: https://doi.org/10.1101/2020.09.22.309229
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High-Resolution, Large Field-of-View, and Multi-View Single Objective Light-Sheet Microscopy
Bin Yang, Alfred Millett-Sikking, Merlin Lange, Ahmet Can Solak, Hirofumi Kobayashi, Andrew York, Loic A. Royer
bioRxiv 2020.09.22.309229; doi: https://doi.org/10.1101/2020.09.22.309229

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