Super-resolution imaging in live cells.

Super-resolution imaging in live cells.
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DOI:
10.1016/j.ydbio.2014.11.025
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发表时间:
2015-05-01
影响因子:
2.7
通讯作者:
Cox, Susan
Cox, Susan
中科院分区:
生物学3区
文献类型:
--
作者:
Cox, Susan

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在过去的二十年里,超分辨率荧光显微镜已经从概念验证实验发展到许多实验室可以使用的商业系统,将分辨率提高了10倍或更多。有三种主要的超分辨率方法,受激辐射耗尽显微镜、结构照明显微镜和定域显微镜,它们都产生了令人惊叹的细胞结构图像。目前的一个主要挑战是优化每种技术的性能,以便相同类型的数据可以常规地在活细胞中获取。有几个主要的挑战,特别是光毒性和获取整个细胞或细胞组的图像的速度。在这篇综述中,我们讨论了可以成功地用于活细胞的各种方法,每种方法在分辨率、速度和实施简易性方面的权衡,以及每种技术可能期望的结果的质量。细胞结构的超分辨率成像可以达到几十纳米的分辨率。主要有三种技术:STED、SIM和定位显微镜。活细胞的超分辨率需要在分辨率、速度和光剂量之间进行权衡。
Over the last twenty years super-resolution fluorescence microscopy has gone from proof-of-concept experiments to commercial systems being available in many labs, improving the resolution achievable by up to a factor of 10 or more. There are three major approaches to super-resolution, stimulated emission depletion microscopy, structured illumination microscopy, and localisation microscopy, which have all produced stunning images of cellular structures. A major current challenge is optimising performance of each technique so that the same sort of data can be routinely taken in live cells. There are several major challenges, particularly phototoxicity and the speed with which images of whole cells, or groups of cells, can be acquired. In this review we discuss the various approaches which can be successfully used in live cells, the tradeoffs in resolution, speed, and ease of implementation which one must make for each approach, and the quality of results that one might expect from each technique. Super-resolution imaging of cell structures can achieve a resolution of tens of nm. There are three major techniques: STED, SIM, and localisation microscopy. Live cell super-resolution requires trading off resolution, speed, and light dose.
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