Three-photon excited fluorescence imaging in neuroscience: From principles to applications

Yujie Xiao; Peng Deng; Yaoguang Zhao; Shasha Yang; Bo Li

doi:10.3389/fnins.2023.1085682

Three-photon excited fluorescence imaging in neuroscience: From principles to applications

Front Neurosci. 2023 Feb 20:17:1085682. doi: 10.3389/fnins.2023.1085682. eCollection 2023.

Authors

Yujie Xiao¹, Peng Deng¹, Yaoguang Zhao¹, Shasha Yang¹, Bo Li¹

Affiliation

¹ State Key Laboratory of Medical Neurobiology, Department of Neurology, Ministry of Education (MOE), Frontiers Center for Brain Science, Institute for Translational Brain Research, Huashan Hospital, Fudan University, Shanghai, China.

Abstract

The development of three-photon microscopy (3PM) has greatly expanded the capability of imaging deep within biological tissues, enabling neuroscientists to visualize the structure and activity of neuronal populations with greater depth than two-photon imaging. In this review, we outline the history and physical principles of 3PM technology. We cover the current techniques for improving the performance of 3PM. Furthermore, we summarize the imaging applications of 3PM for various brain regions and species. Finally, we discuss the future of 3PM applications for neuroscience.

Keywords: deep brain imaging; functional imaging; rodents; structural imaging; three-photon excited fluorescence microscopy.

Publication types

Review

Grants and funding

This work was supported by the National Natural Science Foundation of China (T2222006 and 32200951), Science and Technology Commission of Shanghai Municipality (22JC1403100), Shanghai Municipal Science and Technology Major Project, Natural Science Foundation of Shanghai (21ZR1404600), China Postdoctoral Science Foundation (2022M720801), and the CAS Key Laboratory of Health Informatics, Shenzhen Institute of Advanced Technology (2011DP173015).