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作 者:周日 王晨光 卢革宇[1] ZHOU Ri;WANG Chen-guang;LU Ge-yu(State Key Laboratory of Integrated Optoelectronics,College of Electronic Science and Engineering,Jilin University,Changchun 130012,China)
机构地区:[1]吉林大学电子科学与工程学院集成光电子学国家重点实验室,吉林长春130012
出 处:《中国光学(中英文)》2022年第6期1228-1242,共15页Chinese Optics
基 金:国家自然科学基金(No.61831011,No.62075079)。
摘 要:脂滴是真核细胞中必不可少的一种球形细胞器,与很多细胞生理学过程息息相关。荧光成像技术是观察研究脂滴最有力的工具之一。受光学衍射极限的限制,传统的宽场以及共聚焦显微镜所能达到的成像分辨率约为250 nm左右,这对于观测小脂滴,尤其是新生脂滴(尺寸约30~60 nm)来说是远远不够的。在这种情况下,近年来新兴的各种能够打破衍射极限的超分辨荧光显微镜(如受激发射损耗显微镜、结构光照明显微镜以及光激活定位显微镜等)逐渐吸引了科研人员的兴趣。为了得到高分辨率脂滴荧光图像,除了上述超分辨显微镜之外,还需要具有与之相匹配的高性能荧光探针。本文将简要介绍这几种超分辨显微镜的工作原理,讨论其对荧光探针光物理性质的特殊要求,并进一步系统总结脂滴超分辨成像荧光探针的研究进展。与此同时,本文将分析对比不同超分辨显微镜在脂滴荧光成像方面的优势与不足,并对其发展趋势进行展望。Lipid droplets are a kind of spherical organelle in eukaryotic cells and are relevant to many cellular physiological processes. Fluorescence imaging techniques are one of the most powerful tools to visualize and study lipid droplets. However, conventional wide-field microscopy and confocal microscopy can only provide a resolution of about 250 nm due to the limitation of optical diffraction. This resolution is quite insufficient for visualizing the small lipid droplets, especially the nascent ones(size of about 30~60 nm). Emerging super-resolution microscopes that can break the diffraction limit(such as stimulated emission depletion microscopy, structured illumination microscopy and photoactivated localization microscopy) have gradually attracted much interest in recent years. To obtain high-resolution fluorescence images of lipid droplets, the advanced fluorescent probes which meet the special requirements of the corresponding super-resolution microscopes are highly essential. This review paper will briefly introduce the working principles of various super-resolution microscopes, discuss the special requirements on the photophysical properties of fluorescent probes, and systematically summarize the research progress of super-resolution imaging of lipid droplets by employing these fluorescent probes. Meanwhile, this review will compare the advantages and shortcomings of different super-resolution techniques for lipid droplets imaging, and prospect their future possible trends.
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