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[基于光片的荧光显微镜术:终于揭示了样本的阴暗面]

[Light-sheet based fluorescence microscopy: the dark side of the sample finally revealed].

作者信息

Girard Philippe P, Forget Benoît C

机构信息

Université Paris Descartes, 75270 Paris Cedex 06, France.

出版信息

Med Sci (Paris). 2011 Aug-Sep;27(8-9):753-62. doi: 10.1051/medsci/2011278018. Epub 2011 Aug 31.

Abstract

Light-sheet based fluorescence microscopy (LSM) is an optical technique that becomes more and more popular for multi-view imaging of in vivo sample in its physiological environment. LSM combines the advantages of the direct optical sectioning to the ones of optical tomography by angular scanning. In fact, a thin light-sheet illuminates laterally a section of the sample, thus limiting the effects of photobleaching and phototoxicity only to the plane of interest. The spatial resolution can be improved by combining multiple views obtained along different angle into a single data, leading to a 3D isotropic rendering of the sample. Such an approach provides several advantages in comparison to conventional 3D microscopic techniques: confocal and multiphoton microscopies. It makes LSM an optical tool suited for imaging specimens with a subcellular resolution even inside an embryo and with temporal resolution adapted for real-time monitoring of biological processes.

摘要

基于光片的荧光显微镜(LSM)是一种光学技术,在对处于生理环境中的体内样本进行多视图成像方面越来越受欢迎。LSM通过角度扫描将直接光学切片的优点与光学层析成像的优点结合起来。实际上,一个薄的光片从侧面照亮样本的一个切片,从而将光漂白和光毒性的影响仅限制在感兴趣的平面上。通过将沿不同角度获得的多个视图组合成单个数据,可以提高空间分辨率,从而实现样本的三维各向同性渲染。与传统的三维显微镜技术(共聚焦显微镜和多光子显微镜)相比,这种方法具有几个优点。这使得LSM成为一种光学工具,适合以亚细胞分辨率对胚胎内部的样本进行成像,并具有适用于实时监测生物过程的时间分辨率。

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