Axial resolution and imaging contrast enhancement in inverted light-sheet microscopy by natural illumination modulation

Zhi Wang; Wei Qiao; Tao Jiang; Siqi Chen; Bolin Lu; Kefu Ning; Rui Jin; Hui Gong; Jing Yuan

doi:10.3389/fnins.2022.1032195

Axial resolution and imaging contrast enhancement in inverted light-sheet microscopy by natural illumination modulation

Front Neurosci. 2022 Oct 18:16:1032195. doi: 10.3389/fnins.2022.1032195. eCollection 2022.

Authors

Zhi Wang^{1

2}, Wei Qiao^{1

2}, Tao Jiang³, Siqi Chen^{1

2}, Bolin Lu^{1

2}, Kefu Ning^{1

2}, Rui Jin^{1

2}, Hui Gong^{1

2

3}, Jing Yuan^{1

2

3}

Affiliations

¹ Wuhan National Laboratory for Optoelectronics, Britton Chance Center for Biomedical Photonics, Huazhong University of Science and Technology, Wuhan, China.
² MoE Key Laboratory for Biomedical Photonics, School of Engineering Sciences, Huazhong University of Science and Technology, Wuhan, China.
³ HUST-Suzhou Institute for Brainsmatics, Jiangsu Industrial Technology Research Institute (JITRI) Institute for Brainsmatics, Jiangsu, China.

Abstract

Inverted light-sheet microscopy (ILSM) is widely employed for fast large-volume imaging of biological tissue. However, the scattering especially in an uncleared sample, and the divergent propagation of the illumination beam lead to a trade-off between axial resolution and imaging depth. Herein, we propose naturally modulated ILSM (NM-ILSM) as a technique to improve axial resolution while simultaneously maintaining the wide field-of-view (FOV), and enhancing imaging contrast via background suppression. Theoretical derivations, simulations, and experimental imaging demonstrate 15% axial resolution increases, and fivefold greater image contrast compared with conventional ILSM. Therefore, NM-ILSM allows convenient imaging quality improvement for uncleared tissue and could extend the biological application scope of ILSM.

Keywords: axial resolution enhancement; imaging contrast improvement; large-volume imaging; light-sheet microscopy; natural illumination modulation; optical sectioning microscopy.