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Organic fluorescent probes for stochastic optical reconstruction microscopy (STORM): Recent highlights and future possibilities

机译:随机光学重建显微镜(STORM)的有机荧光探针:近期亮点和未来可能性

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Super-resolution fluorescence imaging by single-molecule localization microscopy (SMLM) offers the possibility of microscopic images with sub-diffraction spatial resolution. Stochastic optical reconstruction microscopy (STORM) is one of the emerging SMLM techniques that has contributed new insights into both the structures and functions of sub-cellular organelles in the cellular context with a spatial resolution virtually at the molecular level. Photo-switching of single fluorophores and position determination are the most common features of this SMLM technique, which allows molecule-resolved information as well as super-resolved images. However, achieving successful STORM-based images relies on the suitable choice of a fluorophore. In particular, the use of ideal organic fluorescent probes has great potential to circumvent common difficulties that arise during the construction of STORM images. However, there is hardly any comprehensive review that critically assesses the criteria for choosing ideal fluorescent probes for STORM and designing new efficient organic fluorescent probes to date. Therefore, this review has particularly focused on the choice of organic fluorescent probes, the essential features for designing new probes and the future prospects for resolving persistent issues in STORM imaging. The utility of organic fluorescent probes in multicolor STORM, 3D STORM and live cell STORM imaging are also discussed to provide a perspective concerning the true application potential of commonly used fluorescent dyes. In this review, we not only describe how organic fluorescent dyes have contributed to the growth of STORM-based super-resolution imaging in eukaryotic biology, but we also attempt to provide a basis on which advanced organic fluorescent probes can be designed and developed in the near future. (C) 2018 Elsevier B.V. All rights reserved.
机译:通过单分子定位显微镜(SMLM)进行的超分辨率荧光成像可提供具有亚衍射空间分辨率的显微图像。随机光学重建显微镜(STORM)是新兴的SMLM技术之一,它为细胞环境中的亚细胞细胞器的结构和功能(在分子水平上具有空间分辨率)提供了新的见识。单个荧光团的光开关和位置确定是此SMLM技术的最常见特征,它允许分子分辨的信息以及超分辨的图像。但是,成功获得基于STORM的图像取决于荧光团的适当选择。特别是,使用理想的有机荧光探针具有很大的潜力来规避在构建STORM图像时出现的常见困难。但是,迄今为止,几乎没有任何全面的评论可以严格地评估为STORM选择理想的荧光探针和设计新的有效有机荧光探针的标准。因此,本综述特别关注有机荧光探针的选择,设计新探针的基本功能以及解决STORM成像中持续存在的问题的未来前景。还讨论了有机荧光探针在多色STORM,3D STORM和活细胞STORM成像中的用途,以提供有关常用荧光染料的实际应用潜力的观点。在这篇综述中,我们不仅描述了有机荧光染料如何促进基于STORM的超分辨率成像在真核生物中的生长,而且我们还尝试提供一个基础,可以在该设计中开发和开发先进的有机荧光探针。不远的将来。 (C)2018 Elsevier B.V.保留所有权利。

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