Viral particle imaging by super-resolution fluorescence microscopy
Viral particle imaging by super-resolution fluorescence microscopy
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DOI：
10.1016/j.chphi.2021.100013
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发表时间：
2021-06-01
影响因子：
2.2
通讯作者：
Boretti, Alberto
中科院分区：
文献类型：
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作者：
Castelletto, Stefania;Boretti, Alberto
荧光显微镜提供了一种简便的成像方法和潜力，在体内成像病毒。然而，它受到光学显微镜衍射极限的限制，该极限高于病毒颗粒的尺寸。荧光超分辨率（FSR）显微镜规避衍射极限。这些技术，荧光显微镜的延伸和替代电子显微镜，建立在过去的二十年，允许可视化的一些病毒成分。在这里，我们总结了不同FSR技术和单一病毒追踪的原理，以及它们当前在病毒学研究中的应用，重点讨论了阻碍它们在活细胞中病毒复制和感染机制研究中广泛使用的一些限制。在病毒学研究和活细胞成像的特定背景下，最知名的FSR方法的优点和缺点的概述，重点放在他们目前的局限性，如标记，成像速度，光漂白，成像深度，以及目前试图克服它们。最近的一些示范，包括不同的超分辨率（SR）的方法的组合提供了答案，作为一个例子，在人类免疫缺陷病毒的复制和从这些经验，可以设想进一步的进展，以揭示其他病毒的开放问题。
Fluorescence microscopy provides a facile imaging methodology and potentials for imaging viruses in vivo. However, it is limited by the diffraction limit of light microscopy which is above the size of the virus particles. Fluorescence super-resolution (FSR) microscopy circumvents the diffraction limit. These techniques, an extension of fluorescence microscopy and alternative to electron microscopy, established over the past twenty years, have permitted the visualization of some virus components. Here, we summarize the principles of different FSR techniques and single virus tracking, and their current applications to virology studies, focusing on some limitations impeding their widespread use in virus replication and infection mechanism studies in living cells. In the specific context of virology studies and live-cell imaging, the advantages and disadvantages of the most known FSR methods are outlined with a focus on their current limitations such as labeling, imaging speed, photo-bleaching, depth of imaging, and the current attempts to overcome them. Some of the recent demonstrations encompassing the combination of different super-resolution (SR) approaches have provided answers to open questions as an example in the human immunodeficiency virus replication and from these experiences, further progress can be envisaged to reveal other viruses.