多色单分子定位显微技术研究进展(特邀)  

Advances in Multicolor Single-molecule Localization Microscopy(Invited)

作　　者：赵悦晗 郝翔[1] ZHAO Yuehan;HAO Xiang(State Key Laboratory of Modern Optical Instrumentation,College of Optical Science and Engineering,Zhejiang University,Hangzhou 310027,China)

机构地区：[1]浙江大学光电科学与工程学院现代光学仪器国家重点实验室,杭州310027

出　　处：《光子学报》2022年第8期301-313,共13页Acta Photonica Sinica

基　　金：中央高校基本科研业务费专项资金(No.226-2022-00137);浙江省“领雁”研发攻关计划(No.2022C01077);浙江省自然科学基金(No.LZ21F050003);国家自然科学基金(No.92050115)。

摘　　要：超分辨显微镜是探测分子尺度的亚细胞结构的有力工具,为生物学研究提供了新的途径。由于许多生物学问题都可以通过分析不同细胞结构间的相互作用进行研究,因此近年来发展了一系列多色超分辨显微技术。本文从生物样品制备和光学系统改进两个角度,对已有的多色单分子定位显微技术进行了总结,简要概括了每种技术的基本原理,分析了每种技术的优缺点及适用范围,重点归纳了近5年通过光学系统改进提升多色单分子定位显微技术性能的各类方法。最后,对快速发展的多色单分子定位显微技术领域进行了展望。Due to its simplicity and minimal invasion,light microscopy has been widely applied in biological science,medicine and chemistry.However,as the result of diffraction,the spatial resolution of standard optical microscopy is limited to about half the wavelength of light,with a lateral dimension of around 200~300 nm and an axial dimension of approximately 500~700 nm.This diffraction limits the ability of light microscopy to resolve the subcellular organization of individual molecules or molecular complexes.To overcome the resolution barrier of optical microscopy caused by diffraction limit,a variety of superresolution microscopy techniques have been developed over the past 15 years,which significantly improved the resolution of light microscopy to 10 nm.These techniques can be roughly classified into two categories:1)single-molecule localization microscopy such as stochastic optical reconstruction microscopy and photoactivation localization microscopy,and 2)spatially patterned illumination-based methods,such as stimulated emission depletion microscopy and structured illumination microscopy.These techniques have revolutionized our understanding of the nanoscale world,Bringing new opportunities and challenges to molecular-scale biological research.This review summarizes the multicolor single-molecule localization microscopy techniques developed in the past five years from the perspectives of biological sample preparation and optical system improvement,The basic principles of each technique are explained and their advantages and disadvantages are discussed.According to the type of fluorescent probes select during sample preparation,some multicolor SMLM can be classified into three categories:Activator-free SMLM technology using synthetic dyes,which requires fluorophores that exhibit similar blinking efficiency under identical photoswitching buffer conditions.techniques using probes with both an activator and a reporter fluorophore enable multicolor SMLM using different activation lasers.And spectrally selective activat

关 键 词：单分子定位 超分辨成像 显微成像 荧光成像 多色成像 

分 类 号：Q63[生物学—生物物理学]