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使用光片显微镜对神经系统发育和功能进行实时成像。

Live imaging of nervous system development and function using light-sheet microscopy.

作者信息

Lemon William C, Keller Philipp J

机构信息

Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia.

出版信息

Mol Reprod Dev. 2015 Jul-Aug;82(7-8):605-18. doi: 10.1002/mrd.22258. Epub 2013 Sep 16.

Abstract

In vivo imaging applications typically require carefully balancing conflicting parameters. Often it is necessary to achieve high imaging speed, low photo-bleaching, and photo-toxicity, good three-dimensional resolution, high signal-to-noise ratio, and excellent physical coverage at the same time. Light-sheet microscopy provides good performance in all of these categories, and is thus emerging as a particularly powerful live imaging method for the life sciences. We see an outstanding potential for applying light-sheet microscopy to the study of development and function of the early nervous system in vertebrates and higher invertebrates. Here, we review state-of-the-art approaches to live imaging of early development, and show how the unique capabilities of light-sheet microscopy can further advance our understanding of the development and function of the nervous system. We discuss key considerations in the design of light-sheet microscopy experiments, including sample preparation and fluorescent marker strategies, and provide an outlook for future directions in the field.

摘要

体内成像应用通常需要仔细平衡相互冲突的参数。通常有必要同时实现高成像速度、低光漂白和光毒性、良好的三维分辨率、高信噪比以及出色的物理覆盖范围。光片显微镜在所有这些方面都具有良好的性能,因此正成为生命科学领域一种特别强大的实时成像方法。我们认为将光片显微镜应用于脊椎动物和高等无脊椎动物早期神经系统的发育和功能研究具有巨大潜力。在这里,我们回顾了早期发育实时成像的最新方法,并展示了光片显微镜的独特能力如何能够进一步推进我们对神经系统发育和功能的理解。我们讨论了光片显微镜实验设计中的关键考虑因素,包括样品制备和荧光标记策略,并对该领域的未来方向进行了展望。

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