Superresolution microscopy for microbiology.

Superresolution microscopy for microbiology.
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DOI:
10.1111/cmi.12024
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发表时间:
2012-12
影响因子:
3.4
通讯作者:
Xiao J
Xiao J
中科院分区:
生物学2区
文献类型:
--
作者:
Coltharp C;Xiao J

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这篇综述从微生物研究的角度对超分辨显微镜进行了介绍。由于细菌细胞的尺寸很小,超分辨率方法特别强大,适用于揭示在传统荧光显微镜下无法分辨的细胞结构细节。在这里,我们描述了三个主要类别的超分辨率光学显微镜背后的方法概念:光激活定位显微镜(PALM)和随机光学重建显微镜(STORM),结构照明显微镜(SIM)和受激发射耗尽(STED)显微镜。然后,我们提出了最近的应用程序,这些技术的微生物系统,揭示了新的构象的细胞结构和描述的新特性,在体内蛋白质的功能和相互作用。最后,我们讨论了与细菌标本实施这些超分辨率技术相关的独特问题,并提出了未来发展的途径。本文的目的是为感兴趣的微生物学家提供必要的技术背景,为他们的生物系统选择合适的超分辨率方法,并介绍设计和分析超分辨率成像实验所需的实际考虑。
This review provides a practical introduction to superresolution microscopy from the perspective of microbiological research. Because of the small sizes of bacterial cells, superresolution methods are particularly powerful and suitable for revealing details of cellular structures that are not resolvable under conventional fluorescence light microscopy. Here we describe the methodological concepts behind three major categories of super-resolution light microscopy: photoactivated localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM), structured illumination microscopy (SIM) and stimulated emission-depletion (STED) microscopy. We then present recent applications of each of these techniques to microbial systems, which have revealed novel conformations of cellular structures and described new properties of in vivo protein function and interactions. Finally, we discuss the unique issues related to implementing each of these superresolution techniques with bacterial specimens and suggest avenues for future development. The goal of this review is to provide the necessary technical background for interested microbiologists to choose the appropriate super-resolution method for their biological systems, and to introduce the practical considerations required for designing and analysing superresolution imaging experiments.
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