Single-scan HiLo with line-illumination strategy for optical section imaging of thick tissues

Wei Qiao; Rui Jin; Tianpeng Luo; Yafeng Li; Guoqing Fan; Qingming Luo; Jing Yuan

doi:10.1364/BOE.419377

Single-scan HiLo with line-illumination strategy for optical section imaging of thick tissues

Biomed Opt Express. 2021 Mar 26;12(4):2373-2383. doi: 10.1364/BOE.419377. eCollection 2021 Apr 1.

Authors

Wei Qiao^{1

2

3}, Rui Jin^{1

2

3}, Tianpeng Luo^{1

2}, Yafeng Li^{1

2}, Guoqing Fan^{1

2}, Qingming Luo^{1

2

4

5}, Jing Yuan^{1

2

4}

Affiliations

¹ Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, China.
² MOE Key Laboratory for Biomedical Photonics, School of Engineering Sciences, Innovation Institute, Huazhong University of Science and Technology, Wuhan 430074, China.
³ These authors contributed equally.
⁴ HUST-Suzhou Institute for Brainsmatics, JITRI Institute for Brainsmatics, Suzhou 215123, China.
⁵ School of Biomedical Engineering, Hainan University, Haikou, China.

Abstract

Optical sectioning has been widely employed for inhibiting out-of-focus backgrounds in three-dimensional (3D) imaging of biological samples. However, point scanning imaging or multiple acquisitions for wide-field optical sectioning in epi-illumination microscopy remains time-consuming for large-scale imaging. In this paper, we propose a single-scan optical sectioning method based on the hybrid illumination (HiLo) algorithm with a line-illumination strategy. Our method combines HiLo background inhibition with confocal slit detection. It thereby offers a higher optical sectioning capability than wide-field HiLo and line-confocal imaging without extra modulation and multiple data acquisition. To demonstrate the optical-sectioning capability of our system, we imaged a thin fluorescent plane and different fluorescence-labeled mouse tissue. Our method shows an excellent background inhibition in thick tissue and thus potentially provides an alternative tool for 3D imaging of large-scale biological tissue.