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• Photonics Research
• Vol. 9, Issue 8, 08001477 (2021)
Rui Jin1, Yalan Yu1、2, Dan Shen1, Qingming Luo1、3、4, Hui Gong1、3、5、*, and Jing Yuan1、3、6、*
Author Affiliations
• 1Britton Chance Center for Biomedical Photonics and MoE Key Laboratory for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, China
• 2Current address: Department of Mechanical and Automation Engineering, Chinese University of Hong Kong, Shatin, Hong Kong
• 3HUST-Suzhou Institute for Brainsmatics, JITRI Institute for Brainsmatics, Suzhou 215123, China
• 4School of Biomedical Engineering, Hainan University, Haikou 570228, China
• 5e-mail: huigong@mail.hust.edu.cn
• 6e-mail: yuanj@hust.edu.cn
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Abstract

Parallel dual-plane imaging with a large axial interval enables the simultaneous observation of biological structures and activities in different views of interest. However, the inflexibility in adjusting the field-of-view (FOV) positions in three dimensions and optical sectioning effects, as well as the relatively small effective axial range limited by spherical aberration, have hindered the application of parallel dual-plane imaging. Herein, we propose a flexible, video-rate, and defocus-aberration-compensated axial dual-line scanning imaging method. We used a stepped mirror to remotely generate and detect dual axial lines with compensation for spherical aberration and FOV-jointing to rearrange into a head-to-head line for high-speed optical sectioning acquisition. The lateral and axial positions of the two FOVs could be flexibly adjusted before and during imaging, respectively. The method also allows the adjustment of optical sectioning effects according to specific experimental requirements. We experimentally verified the consistent imaging performance over an axial range of 300 μm. We demonstrated high throughput by simultaneously imaging Brownian motions in two $250 μm×250 μm$ FOVs with axial and lateral intervals of 150 μm and 240 μm, respectively, at 24.5 Hz. We also showed potential application in functional imaging by simultaneously acquiring neural activities in the optic tectum and hindbrain of a zebrafish brain. The proposed method is, thus, advantageous compared to existing parallel dual-plane imaging and potentially facilitates intravital biological study in large axial range.
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Rui Jin, Yalan Yu, Dan Shen, Qingming Luo, Hui Gong, Jing Yuan. Flexible, video-rate, and aberration-compensated axial dual-line scanning imaging with field-of-view jointing and stepped remote focusing[J]. Photonics Research, 2021, 9(8): 08001477