Axially overlapped multi-focus light sheet with enlarged field of view

Li, H; Wu, Z; Yang, Z; Zhanghao, K; Xi, P; Jin, D

Axially overlapped multi-focus light sheet with enlarged field of view

Li, H Wu, Z Yang, Z Zhanghao, K Xi, P Jin, D

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Publisher:: American Institute of Physics
Publication Type:: Journal Article
Citation:: Applied Physics Letters, 2021, 118, (22), pp. 1-6
Issue Date:: 2021-05-31

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Field	Value	Language
dc.contributor.author	Li, H
dc.contributor.author	Wu, Z
dc.contributor.author	Yang, Z
dc.contributor.author	Zhanghao, K
dc.contributor.author	Xi, P
dc.contributor.author	Jin, D https://orcid.org/0000-0003-1046-2666
dc.date.accessioned	2022-01-15T22:42:30Z
dc.date.available	2022-01-15T22:42:30Z
dc.date.issued	2021-05-31
dc.identifier.citation	Applied Physics Letters, 2021, 118, (22), pp. 1-6
dc.identifier.issn	0003-6951
dc.identifier.issn	1077-3118
dc.identifier.uri	http://hdl.handle.net/10453/153156
dc.description.abstract	Light sheet fluorescence microscopy provides optical sectioning and is widely used in volumetric imaging of large specimens. However, the axial resolution and the lateral Field of View (FoV) of the system, defined by the light sheet, typically limit each other due to the spatial band product of the excitation objective. Here, we develop a simple multi-focus scheme to extend the FoV, where a Gaussian light sheet can be focused at three or more consecutive positions. Axially overlapped multiple light sheets significantly enlarge the FoV with improved uniformity and negligible loss in axial resolution. By measuring the point spread function of fluorescent beads, we demonstrated that the obtained light sheet has a FoV of 450 μm and a maximum axial FWHM of 7.5 μm. Compared with the conventional single-focus one, the multi-focus Gaussian light sheet displays a significantly improved optical sectioning ability over the full FoV when imaging cells and zebrafish. Light sheet fluorescence microscopy (LSFM) has become an indispensable tool in volumetric imaging, with the advances in high spatiotemporal resolution and low photo-toxicity to the fluorescent sample. The open-source design with detailed instructions encourages DIY setups, which has significantly accelerated the wide-range adoption and applications of LSFM. Pioneering works, including OpenSPIM,1 OpenSpin,2 and the recent mesoSPIM,3 provide detailed protocols for building and using the microscopes. These joint efforts further allow the biology labs to build their LSFM system for a specific application, including more complex schemes, such as multiview excitation or detection configurations.4–
dc.language	en
dc.publisher	American Institute of Physics
dc.relation	National Natural Science Foundation of China61729501
dc.relation.ispartof	Applied Physics Letters
dc.relation.isbasedon	10.1063/5.0049013
dc.rights	info:eu-repo/semantics/openAccess
dc.subject	02 Physical Sciences, 09 Engineering, 10 Technology
dc.subject.classification	Applied Physics
dc.title	Axially overlapped multi-focus light sheet with enlarged field of view
dc.type	Journal Article
utslib.citation.volume	118
utslib.for	02 Physical Sciences
utslib.for	09 Engineering
utslib.for	10 Technology
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Mathematical and Physical Sciences
pubs.organisational-group	/University of Technology Sydney/Strength - IBMD - Initiative for Biomedical Devices
utslib.copyright.status	open_access	*
pubs.consider-herdc	false
dc.date.updated	2022-01-15T22:42:28Z
pubs.issue	22
pubs.publication-status	Published
pubs.volume	118
utslib.citation.issue	22

Abstract:

Light sheet fluorescence microscopy provides optical sectioning and is widely used in volumetric imaging of large specimens. However, the axial resolution and the lateral Field of View (FoV) of the system, defined by the light sheet, typically limit each other due to the spatial band product of the excitation objective. Here, we develop a simple multi-focus scheme to extend the FoV, where a Gaussian light sheet can be focused at three or more consecutive positions. Axially overlapped multiple light sheets significantly enlarge the FoV with improved uniformity and negligible loss in axial resolution. By measuring the point spread function of fluorescent beads, we demonstrated that the obtained light sheet has a FoV of 450 μm and a maximum axial FWHM of 7.5 μm. Compared with the conventional single-focus one, the multi-focus Gaussian light sheet displays a significantly improved optical sectioning ability over the full FoV when imaging cells and zebrafish. Light sheet fluorescence microscopy (LSFM) has become an indispensable tool in volumetric imaging, with the advances in high spatiotemporal resolution and low photo-toxicity to the fluorescent sample. The open-source design with detailed instructions encourages DIY setups, which has significantly accelerated the wide-range adoption and applications of LSFM. Pioneering works, including OpenSPIM,1 OpenSpin,2 and the recent mesoSPIM,3 provide detailed protocols for building and using the microscopes. These joint efforts further allow the biology labs to build their LSFM system for a specific application, including more complex schemes, such as multiview excitation or detection configurations.4–

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http://hdl.handle.net/10453/153156