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Extended field of view of light-sheet fluorescence microscopy by scanning multiple focus-shifted Gaussian beam arrays

Authors :
Yuan Zhou
Shaohui Yan
Chen Bai
Junwei Min
Xing Li
Xianghua Yu
Chao Liu
Yanlong Yang
Baoli Yao
Dan Dan
Source :
Optics express. 29(4)
Publication Year :
2021

Abstract

Light-sheet fluorescence microscopy (LSFM) facilitates high temporal-spatial resolution, low photobleaching and phototoxicity for long-term volumetric imaging. However, when a high axial resolution or optical sectioning capability is required, the field of view (FOV) is limited. Here, we propose to generate a large FOV of light-sheet by scanning multiple focus-shifted Gaussian beam arrays (MGBA) while keeping the high axial resolution. The positions of the beam waists of the multiple Gaussian beam arrays are shifted in both axial and lateral directions in an optimized arranged pattern, and then scanned along the direction perpendicular to the propagation axis to form an extended FOV of light-sheet. Complementary beam subtraction method is also adopted to further improve axial resolution. Compared with the single Gaussian light-sheet method, the proposed method extends the FOV from 12 μm to 200 μm while sustaining the axial resolution of 0.73 μm. Both numerical simulation and experiment on samples are performed to verify the effectiveness of the method.

Details

ISSN :
10944087
Volume :
29
Issue :
4
Database :
OpenAIRE
Journal :
Optics express
Accession number :
edsair.doi.dedup.....d9cb19dd416e128c080b850719846d4a