关键生物信号枢纽的细胞原位单分子定位超高分辨率解析  

In Situ Single⁃Molecule Localization⁃Based Super⁃Resolution Analysis of Key Cellular Signalosomes

作　　者：林健 陈鑫[1] Lin Jian;Chen Xin(School of Life Sciences,Xiamen University,Xiamen 361102,Fujian,China)

机构地区：[1]厦门大学生命科学学院,福建厦门361102

出　　处：《中国激光》2024年第3期20-36,共17页Chinese Journal of Lasers

基　　金：国家自然科学基金面上项目(32370803,32070736,31871386);国家自然科学基金青年项目(31501115);福建省自然科学基金杰青项目(2023J06003);厦门大学校长基金(20720210114);厦门大学拔尖学生贵重实验仪器设备开放创新基金(KFJJ-202206,KFJJ-202316)。

摘　　要：细胞内存在大量性质各异的大分子复合物,它们是控制生命活动的核心信号枢纽。荧光显微成像因其分子特异性、细胞原位可视化和多色标记解析等优势,已成为当今生物学研究不可或缺的技术手段。而突破光学衍射极限的超分辨光学成像技术为进一步理解多种重要生物百纳米信号体的原位结构和功能调节提供了亚细胞尺度甚至单分子精度的研究思路和方案。首先阐述了超分辨成像和单分子定位显微镜的基本原理,介绍了近年来在多项生命科学研究中的代表性应用案例,结合实际研究经验总结了超分辨光学成像技术在使用中面临的诸多挑战和未来发展方向,提出了基于原位纳米成像解析信号枢纽组织结构将有力促进新型疾病治疗靶点和策略的开发。Significance Cells,the basic structural and functional units,play an essential role in the development,aging,disease,and death of organisms.Since the first microscopic observation of cells by Robert Hooke in 1665,numerous advanced technical and theoretical methods have been developed over the past centuries to microscopically visualize cells,enabling a thorough analysis of life activities from the cellular to the molecular levels.Cells are composed of numerous macromolecular complexes with different sizes and diverse compositions.For example,the eukaryotic 80S ribosome is composed of large and small subunits,and each subunit contains various ribosomal RNA(rRNA)and ribosomal proteins.These complexes are usually considered as core signaling hubs to precisely control cellular structures and functions during various biological activities.Accordingly,cells can properly generate immediate responses to frequent environmental changes and distinct cellular stresses.Therefore,a mechanistic investigation of the structural assembly of these key signaling hubs and their functional regulation is necessary to improve our understanding of life activities and to identify potential therapeutic targets for disease treatment.Currently,single-particle cryo-electron microscope(cryo-EM),which requires only a small number of samples for analysis and does not involve the use of crystals unlike traditional X-ray-based methods,is the most powerful tool in structural biology owing to its extremely high spatial resolution.However,precisely resolving a structure using cryo-EM involves purification or enrichment of the target biomolecules,which increases the risk of inconsistency between in vitro resolved structures and the native structures in cells.Notably,owing to the lack of molecular specificity,understanding the interactions among different molecules when resolving multicomponent complexes is challenging.In addition,high-quality cryo-EM analyses depend on the computational averaging of thousands of images of identical particles with good

关 键 词：生物光学 超分辨成像 单分子定位 生物信号体 信号转导 

分 类 号：O436[机械工程—光学工程] Q2-33[理学—光学]