Chai Yuming, Qi Kexin, Wu Yubin, Li Daguang, Tan Guodong, Guo Yuqi, Chu Jun, Mu Yu, Shen Chen, Wen Quan
Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China.
Hefei National Research Center for Physical Sciences at the Microscale, Center for Integrative Imaging, University of Science and Technology of China, Hefei, China.
iScience. 2023 Nov 3;27(1):108385. doi: 10.1016/j.isci.2023.108385. eCollection 2024 Jan 19.
We introduce an all-optical technique that enables volumetric imaging of brain-wide calcium activity and targeted optogenetic stimulation of specific brain regions in unrestrained larval zebrafish. The system consists of three main components: a 3D tracking module, a dual-color fluorescence imaging module, and a real-time activity manipulation module. Our approach uses a sensitive genetically encoded calcium indicator in combination with a long Stokes shift red fluorescence protein as a reference channel, allowing the extraction of Ca activity from signals contaminated by motion artifacts. The method also incorporates rapid 3D image reconstruction and registration, facilitating selective optogenetic stimulation of different regions of the brain. By demonstrating that selective light activation of the midbrain regions in larval zebrafish could reliably trigger biased turning behavior and changes of brain-wide neural activity, we present a valuable tool for investigating the causal relationship between distributed neural circuit dynamics and naturalistic behavior.
我们介绍了一种全光学技术，该技术能够对自由游动的斑马鱼幼体进行全脑钙活性的体积成像，并对特定脑区进行靶向光遗传学刺激。该系统由三个主要部分组成：一个三维跟踪模块、一个双色荧光成像模块和一个实时活动操纵模块。我们的方法使用一种灵敏的基因编码钙指示剂，并结合一种具有长斯托克斯位移的红色荧光蛋白作为参考通道，从而能够从受运动伪影污染的信号中提取钙活性。该方法还结合了快速三维图像重建和配准，便于对大脑不同区域进行选择性光遗传学刺激。通过证明对斑马鱼幼体中脑区域的选择性光激活能够可靠地触发偏向性转向行为和全脑神经活动的变化，我们展示了一种用于研究分布式神经回路动力学与自然行为之间因果关系的有价值工具。
