Synchrotron microCT imaging of soft tissue in juvenile zebrafish reveals retinotectal projections.
Synchrotron microCT imaging of soft tissue in juvenile zebrafish reveals retinotectal projections.
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幼年斑马鱼软组织的同步加速器 microCT 成像揭示了视网膜顶盖投影。
DOI：
10.1117/12.2267477
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发表时间：
2017
期刊：
影响因子：
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通讯作者：
Cheng,KeithC
中科院分区：
文献类型：
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作者：
Xin,Xuying;Clark,Darin;Ang,KhaiChung;vanRossum,DamianB;Copper,Jean;Xiao,Xianghui;LaRiviere,PatrickJ;Cheng,KeithC
生物医学研究和临床诊断将受益于解剖表型的全体积测定。形态学表型分析的综合工具是表型组学新兴领域的核心，这需要从受遗传、疾病或环境变量影响的生物体中收集高通量、系统、准确和可重复的数据。理论上，完整的解剖表型需要对整个生物体中的每种细胞类型进行评估，但由于缺乏一种公正的3D成像方法，这种理想目前是站不住脚的，这种方法允许对任何细胞类型进行组织病理学评估，尽管光学不透明。组织病理学是目前诊断表型的临床标准，涉及组织切片的显微研究，以评估组织结构、疾病机制和生理状态的定性方面。然而，组织结构的定量特征，如细胞组成和组织体积中的细胞计数，由于组织切片的特点，包括不完整的采样和5微米厚组织板的二维成像的限制，只能近似地估计。我们使用了一种小型的脊椎动物，斑马鱼，来测试microCT在系统的宏观和微观形态表型上的潜力。虽然细胞分辨率通常是通过光片荧光显微镜和光学断层扫描等方法实现的，但这些方法不能提供组织学特征的全细胞视角，并且受到可见光通过着色和不透明标本的有限穿透的限制，这是斑马鱼幼鱼的特征。在这里，我们提供了一个神经解剖学的例子，可以通过1.43微米各向同性体素分辨率的染色软组织微ct进行研究。我们得出的结论是，同步微ct是一种3D成像形式，可能潜在地用于光学不透明组织的更可复制，大规模，形态表型。软组织微ct的进一步发展、可视化和定量工具的开发将增强其实用性。
Biomedical research and clinical diagnosis would benefit greatly from full volume determinations of anatomical phenotype. Comprehensive tools for morphological phenotyping are central for the emerging field of phenomics, which requires high-throughput, systematic, accurate, and reproducible data collection from organisms affected by genetic, disease, or environmental variables. Theoretically, complete anatomical phenotyping requires the assessment of every cell type in the whole organism, but this ideal is presently untenable due to the lack of an unbiased 3D imaging method that allows histopathological assessment of any cell type despite optical opacity. Histopathology, the current clinical standard for diagnostic phenotyping, involves the microscopic study of tissue sections to assess qualitative aspects of tissue architecture, disease mechanisms, and physiological state. However, quantitative features of tissue architecture such as cellular composition and cell counting in tissue volumes can only be approximated due to characteristics of tissue sectioning, including incomplete sampling and the constraints of 2D imaging of 5 micron thick tissue slabs. We have used a small, vertebrate organism, the zebrafish, to test the potential of microCT for systematic macroscopic and microscopic morphological phenotyping. While cell resolution is routinely achieved using methods such as light sheet fluorescence microscopy and optical tomography, these methods do not provide the pancellular perspective characteristic of histology, and are constrained by the limited penetration of visible light through pigmented and opaque specimens, as characterizes zebrafish juveniles. Here, we provide an example of neuroanatomy that can be studied by microCT of stained soft tissue at 1.43 micron isotropic voxel resolution. We conclude that synchrotron microCT is a form of 3D imaging that may potentially be adopted towards more reproducible, large-scale, morphological phenotyping of optically opaque tissues. Further development of soft tissue microCT, visualization and quantitative tool development will enhance its utility.