低功率受激辐射损耗超分辨显微成像技术研究进展及展望  

Research Progress and Prospects of Low⁃Power Stimulated Emission Depletion Microscopy

作　　者：周浩贤 王璐玮 张仁龙 林方睿 刘丽炜 屈军乐[1] Zhou Haoxian;Wang Luwei;Zhang Renlong;Lin Fangrui;Liu Liwei;Qu Junle(Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education/Guangdong Province,College of Physics and Optoelectronic Engineering,Shenzhen University,Shenzhen 518060,Guangdong,China)

机构地区：[1]深圳大学物理与光电工程学院,光电子器件与系统教育部/广东省重点实验室,广东深圳518060

出　　处：《中国激光》2024年第21期1-18,共18页Chinese Journal of Lasers

基　　金：国家自然科学基金(62127819);广东省基础与应用基础研究基金(2024A1515030193,2023A1515010795);深圳市光子学与生物光子学重点实验室资助项目(ZDSYS20210623092006020);深圳市科技计划资助项目(JCYJ20220818100202005)。

摘　　要：受激辐射损耗(STED)是一种功能强大的远场超分辨显微成像技术,已被广泛用于细胞和组织切片等生物样品的超分辨成像。通过增加损耗激光的功率可以显著提高STED超分辨成像的空间分辨率和成像深度,然而,过高的激光功率会引起严重的光漂白及光毒性。因此,如何在保证成像质量的同时有效降低STED超分辨成像所需的损耗激光强度,是目前STED技术在生物成像领域面临的关键挑战。本文从STED成像的基本原理出发,分别从STED探针、单分子定位、图像处理和时间分辨探测等4个方面探讨了实现低功率STED超分辨成像的策略。针对以上4种策略及其优缺点的深入分析,为STED技术在生物学领域的应用提供了有价值的参考和指导。Significance Known for its non-invasive and non-destructive nature,optical microscopy can provide structural and functional insights into biological specimens,thus driving progress in fields such as biology,medicine,and related disciplines.Over the past four centuries,optical microscopy has witnessed significant developments.These have been particularly accelerated in the last century by technological advancements in lasers and computational methods.These advancements have led to revolutionary changes,making optical microscopy an essential tool in critical sectors such as healthcare,education,and food safety.With the increasing exploration in cellular biology and biomedicine,a growing need has arisen for optical microscopes with molecular or nanoscale spatial resolution,as exemplified by super-resolution optical microscopy(SRM).Of the various SRM techniques,stimulated emission depletion(STED)microscopy stands out because it achieves resolution enhancement by modulating the depletion power relative to the redshift in the excitation wavelength in the imaging setup.However,excessive power depletion poses challenges,including photobleaching of fluorophores and phototoxicity to biological specimens,which constrain the utility of STED in live-cell imaging scenarios.In recent years,researchers worldwide have collaborated to advance the field of STED microscopy with a particular focus on developing strategies to reduce depletion power.This effectively decreases the amount of power required for imaging while maintaining resolution accuracy.These studies are crucial for understanding the intricate details and underlying mechanisms in living organisms.Progress In this review,we discuss the basic principles of STED microscopy and emphasize its crucial role in achieving superresolution imaging of biological samples.Achieving super-resolution imaging using STED microscopy requires precise control over the spatial,temporal,and spectral aspects(Fig.1).By applying the theoretical framework that governs the resolution calculations

关 键 词：超分辨成像 受激辐射损耗 损耗光功率 纳米探针 荧光寿命 

分 类 号：TH742[机械工程—光学工程]