[논문]3D Light-Sheet Fluorescence Microscopy of Cranial Neurons and Vasculature during Zebrafish Embryogenesis

Park, Ok Kyu; Kwak, Jina; Jung, Yoo Jung; Kim, Young Ho; Hong, Hyun-Seok; Hwang, Byung Joon; Kwon, Seung-Hae; Kee, Yun

doi:10.14348/molcells.2015.0160

3D Light-Sheet Fluorescence Microscopy of Cranial Neurons and Vasculature during Zebrafish Embryogenesis 원문보기

Molecules and cells, v.38 no.11, 2015년, pp.975 - 981

Park, Ok Kyu (Korea Basic Science Institute Chuncheon Center) , Kwak, Jina (Department of Systems Immunology, College of Biomedical Science, Kangwon National University) , Jung, Yoo Jung (Department of Systems Immunology, College of Biomedical Science, Kangwon National University) , Kim, Young Ho (Medifron_DBT, Inc.) , Hong, Hyun-Seok (Medifron_DBT, Inc.) , Hwang, Byung Joon (Department of Molecular Bioscience, College of Biomedical Science, Kangwon National University) , Kwon, Seung-Hae (Korea Basic Science Institute Chuncheon Center) , Kee, Yun (Department of Systems Immunology, College of Biomedical Science, Kangwon National University)

Abstract ▼ AI-Helper

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.

주제어

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문제 정의

S4B). This study demonstrates that light-sheet imaging allows the complete visualization of blood cell formation, local distribution and circulation through developing blood vessels in the entire zebrafish embryo during development.

제안 방법

We also imaged circulating blood cells within blood vessels of the whole anterior embryonic body in 3D. Our results reveal details of the spatial relationship between the developing nervous system and blood vessels, as well as providing a direct visualization of blood flow through the entire cranial vasculature. Such data cannot be obtained by traditional point scanning microscopy.
This study is the first to visualize and comprehensively map the 3D anatomy of cranial neurons, vasculature tissues and blood cells in the entire embryonic brains at different stages of embryogenesis using Light-Sheet microscopy. SPIM overcomes many of the limitations of focal imaging, including confocal microscopy, generating live 3D images at high speed and spatio-temporal resolution.

대상 데이터

Data is comprised of 479 optical sections of 1.176 μm thickness using optimum interval option.
Data is comprised of1740 optical sections of 0.488 μm thickness.

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