3D Light-Sheet Fluorescence Microscopy of Cranial Neurons and Vasculature during Zebrafish Embryogenesis
2015
Park, O.K., Korea Basic Science Institute Chuncheon Center, Chuncheon, Republic of Korea | Kwak, J., Kangwon National University, Chuncheon, Chuncheon, Republic of Korea | Jung, Y.J., Kangwon National University, Chuncheon, Chuncheon, Republic of Korea | Kim, Y.H., Medifron DBT, Inc., Ansan, Republic of Korea | Hong, H.S., Medifron DBT, Inc., Ansan, Republic of Korea | Hwang, B.J., Kangwon National University, Chuncheon, Republic of Korea | Kwon, S.H., Korea Basic Science Institute Chuncheon Center, Chuncheon, Republic of Korea | Kee, Y., Kangwon National University, Chuncheon, Chuncheon, Republic of Korea
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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