3D Light-Sheet Fluorescence Microscopy of Cranial Neurons and Vasculature during Zebrafish Embryogenesis |
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| Authors: | Ok Kyu Park Jina Kwak Yoo Jung Jung Young Ho Kim Hyun-Seok Hong Byung Joon Hwang Seung-Hae Kwon Yun Kee |
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| Institution: | 1.Korea Basic Science Institute Chuncheon Center, Chuncheon 200-701, Korea;2.Department of Systems Immunology, College of Biomedical Science, Kangwon National University, Chuncheon 200-701, Korea;3.Medifron_DBT, Inc., Ansan 425-839, Korea;4.Department of Molecular Bioscience, College of Biomedical Science, Kangwon National University, Chuncheon 200-701, Korea;5.Institute of Bioscience and Biotechnology, Kangwon National University, Chuncheon 200-701, Korea |
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| Abstract: | Precise 3D spatial mapping of cells and their connections within living tissues is required to fully understand developmental processes and neural activities. Zebrafish embryos are relatively small and optically transparent, making them the vertebrate model of choice for live in vivo imaging. However, embryonic brains cannot be imaged in their entirety by confocal or two-photon microscopy due to limitations in optical range and scanning speed. Here, we use light-sheet fluorescence microscopy to overcome these limitations and image the entire head of live transgenic zebrafish embryos. We simultaneously imaged cranial neurons and blood vessels during embryogenesis, generating comprehensive 3D maps that provide insight into the coordinated morphogenesis of the nervous system and vasculature during early development. In addition, blood cells circulating through the entire head, vagal and cardiac vasculature were also visualized at high resolution in a 3D movie. These data provide the foundation for the construction of a complete 4D atlas of zebrafish embryogenesis and neural activity. |
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| Keywords: | 3D reconstruction light-sheet microscopy neurons SPIM vasculature zebrafish embryos |
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