| 多种超分辨荧光成像技术比较和进展评述 |
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| 引用本文： | 陈婕,刘文娟,徐兆超. 多种超分辨荧光成像技术比较和进展评述[J]. 色谱, 2021, 39(10): 1055-1064. DOI: 10.3724/SP.J.1123.2021.06015 |
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| 作者姓名： | 陈婕 刘文娟 徐兆超 |
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| 作者单位： | 1.中国科学院分离分析化学重点实验室, 中国科学院大连化学物理研究所, 辽宁 大连 1160232.中国科学院大学, 北京 100049 |
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| 基金项目： | 国家自然科学基金(21878286);国家自然科学基金(22078314) |
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| 摘 要： | 所见即所得是生命科学研究的中心哲学,贯穿在不断认识单个分子、分子复合体、分子动态行为和整个分子网络的历程中.活的动态的分子才是有功能的,这决定了荧光显微成像在生命科学研究中成为不可替代的工具.但是当荧光成像聚焦到分子水平的时候,所见并不能给出想要得到的.这个障碍是由于受光学衍射极限的限制,荧光显微镜无法在衍射受限的空间...
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| 关 键 词： | 超分辨荧光成像 纳米尺度 可视化 活细胞结构和动态 |
| 收稿时间： | 2021-06-09 |
Comparison and progress review of various super-resolution fluorescence imaging techniques |
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| CHEN Jie,LIU Wenjuan,XU Zhaochao. Comparison and progress review of various super-resolution fluorescence imaging techniques[J]. Chinese journal of chromatography, 2021, 39(10): 1055-1064. DOI: 10.3724/SP.J.1123.2021.06015 |
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| Authors: | CHEN Jie LIU Wenjuan XU Zhaochao |
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| Affiliation: | 1. CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China2. University of Chinese Academy of Sciences, Beijing 100049, China |
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| Abstract: | “Seeing is believing” is the central philosophy of life science research, which runs through the continuous understanding of individual molecules, molecular complexes, molecular dynamic behavior, and the entire molecular network. Living and dynamic molecules are functional in nature; therefore, fluorescence microscopy has emerged as an irreplaceable tool in life science research. However, when fluorescence imaging is performed at the molecular level, some artificial signals may lead to erroneous experimental results. This obstacle is due to the limitation of the optical diffraction limit, and the fluorescence microscope cannot distinguish the target in the diffraction-limited space. Super-resolution fluorescence imaging technology breaks through the diffraction limit, allows visualization of biomolecules at the nanometer scale to the single-molecule level, and allows us to study the structure and dynamic processes of living cells with unprecedented spatial and temporal resolution. It has become a powerful tool for life science research and is gradually being applied to material science, catalytic reaction processes, and photolithography as well. The principle of super-resolution imaging technologies is different; therefore, it has different technical performances, thus limiting their specific technical characteristics and application scope. Current mainstream super-resolution imaging technologies can be classified into three types: structured illumination microscopy (SIM), stimulated emission depletion (STED), and single-molecule localization microscopy (SMLM). These microscopes use different complex technologies, but the strategy is the same and simple, i.e. two adjacent luminous points in a diffraction-limited space can be spatially resolved by time resolution. SIM has been used for three-dimensional real-time imaging in multicellular organisms; however, compared with other technologies, its lower horizontal and vertical resolutions need to be further optimized. STED is limited by its small imaging field of view and high photobleaching; however, the best time resolution can be considered at a high spatial resolution, and it has been proven that three-color STED imaging can be performed. In SMLM super-resolution imaging, the time resolution is affected by the time required to locate all fluorophores, which is closely related to the switching and luminescence properties of the fluorophore. With the improvement in horizontal and vertical resolution of imaging, the image acquisition speed, photobleaching characteristics, and the possibility of multi-color and dynamic imaging have increasingly become the key determinants of super-resolution fluorescence imaging. Thus far, the main use of super-resolution imaging technology has been focused on biological applications for studying structural changes less than 200 nm in dimension. In addition to the combination of structural and morphological characterization with biomolecular detection and identification, super-resolution imaging technology is rapidly expanding into the fields of interaction mapping, multi-target detection, and real-time imaging. In the latter applications, super-resolution imaging technology is particularly advantageous because of more flexible sample staining, higher labeling efficiency, faster and simpler readings, and gentler sample preparation procedures. In this article, we compare the principles of these three technologies and introduce their application progress in biology. We expect the results described herein will help researchers clarify the technical advantages and applicable application directions of different super-resolution imaging technologies, thus facilitating researchers in making reasonable choices in future research. |
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| Keywords: | super-resolution fluorescence imaging nanometer scale visualization cellular structure and dynamics |
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