All-optical interrogation of brain-wide activity in freely swimming larval zebrafish
Yuming Chai,
Kexin Qi,
Yubin Wu,
Daguang Li,
Guodong Tan,
Yuqi Guo,
Jun Chu,
Yu Mu,
Chen Shen,
Quan Wen
Affiliations
Yuming Chai
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; Corresponding author
Kexin Qi
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
Yubin Wu
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
Daguang Li
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
Guodong Tan
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
Yuqi Guo
Guangdong Provincial Key Laboratory of Biomedical Optical Imaging Technology and Center for Biomedical Optics and Molecular Imaging, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China
Jun Chu
Guangdong Provincial Key Laboratory of Biomedical Optical Imaging Technology and Center for Biomedical Optics and Molecular Imaging, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China
Yu Mu
Institute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai, China
Chen Shen
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; Corresponding author
Quan Wen
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; Corresponding author
Summary: 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 Ca2+ activity from signals contaminated by motion artifacts. The method also incorporates rapid 3D image reconstruction and registration, facilitating real-time 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.
