Laboratory of Cellular Pharmacology, Graduate School of Pharmaceutical Sciences, Nagoya University, Nagoya, Japan.
RIKEN Center for Advanced Photonics, Saitama, Japan; Department of Advanced Imaging, Graduate School of Biostudies, Kyoto University, Kyoto, Japan; PRESTO/CREST, Japan Science and Technology Agency, Saitama, Japan.
Neurosci Res. 2022 Jun;179:15-23. doi: 10.1016/j.neures.2022.03.007. Epub 2022 Mar 31.
Monitoring neural activity and associating neural dynamics with the anatomical connectome are required to understand how the brain works. Neural dynamics are measured by electrophysiology and optical imaging. Since the discovery of the two-photon excitation phenomenon, significant progress has been made in deep imaging for capturing neural activity from numerous neurons in vivo. The development of two-photon microscopy is aimed to image neural activity from a large and deep region with high spatial (x, y, and z) and temporal (t) resolutions at a high signal-to-noise ratio. Imaging deep regions along the optical axis (z-axis) is particularly challenging because heterogeneous biological tissues scatter and absorb light. Recent advances in the light focus modulation technology at high speeds in three dimensions (x, y, and z) have allowed multiplane two-photon imaging. z-Focus control by varifocal optical systems, such as ferroelectric liquid lenses, gradient refractive index lenses, and adaptive optical element systems, and multiplexing by time- and wavelength-division strategies have allowed to rapidly observe specimens at different focal depths. Herein, we overview the recent advances in multiplane functional imaging systems that enable four-dimensional (x, y, z, and t) analysis of neural dynamics, with a special emphasis on z-scanning mechanisms and multiplexing strategies.
监测神经活动并将神经动力学与解剖连接组学关联起来，是理解大脑工作原理所必需的。神经动力学通过电生理学和光学成像来测量。自双光子激发现象发现以来，在捕获体内大量神经元的神经活动的深层成像方面取得了重大进展。双光子显微镜的发展旨在以高信噪比在大而深的区域内以高空间（x、y 和 z）和时间（t）分辨率成像神经活动。沿着光轴（z 轴）对深部区域进行成像特别具有挑战性，因为异质生物组织会散射和吸收光。近年来，在三维（x、y 和 z）高速光焦点调制技术方面的进展使得多层面双光子成像成为可能。通过可变焦光学系统（如铁电液晶透镜、梯度折射率透镜和自适应光学元件系统）进行 z 焦点控制，以及通过时分和波分复用策略进行复用，使得能够快速观察不同焦点深度的标本。在此，我们概述了多层面功能成像系统的最新进展，这些系统能够对神经动力学进行四维（x、y、z 和 t）分析，特别强调 z 扫描机制和复用策略。
