受激辐射损耗超分辨显微成像系统研究的新进展  

New advances in the research of stimulated emission depletion super-resolution microscopy

作　　者：王佳林[1] 严伟[1] 张佳 王璐玮 杨志刚 屈军乐[1] Wang Jia-Lin;Yan Wei;Zhang Jia;Wang Lu-Wei;Yang Zhi-Gang;Qu Jun-Le(Optoelectronic Device and System,Ministry of Education/Key Laboratory of Guangdong Province,College of Physics and Optoelectronic Engineering,Shenzhen University,Shenzhen 518060,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,深圳518060

出　　处：《物理学报》2020年第10期263-284,共22页Acta Physica Sinica

基　　金：国家重点基础研究发展计划(批准号:2017YFA0700500);国家自然科学基金(批准号:61975127,61525503,61620106016,61835009,81727804);广东省高等学校科技创新(重点)项目(批准号:2016KCXTD007);广东省自然科学基金创新团队项目(批准号:2014A030312008);深圳市基础研究项目(批准号:JCYJ2018030512304883,JCY20170818100153423)资助的课题.

摘　　要：由于受到衍射极限的影响,传统光学显微镜的分辨率被限制在半个波长左右.近二十年来出现了许多通过不同方法绕过光学衍射极限的超分辨成像技术,其中,受激辐射损耗显微(stimulated emission depletion microscopy,STED)通过引入一束环形损耗光来抑制荧光光斑外围荧光分子的发光,以达到减小点扩散函数的目的,实现超分辨成像.经过近些年的发展,STED系统无论从光束的产生、校准和扫描,还是最后的成像,都有了很大的发展.本文将简要介绍STED成像技术的基本原理,详述STED超分辨成像技术出现至今在光源、扫描及成像系统等方面的进展,以及在三维成像和多色成像方面的发展现状,STED技术与其他显微技术的结合.最后,本文对STED技术近几年的研究新进展进行了系统的论述,对STED技术未来的发展趋势进行了探讨.Due to the influence of the diffraction limit,the lateral spatial resolution and axial spatial resolution of traditional optical microscopes are limited to ~200 nm and ~500 nm,respectively.In the past two decades,with the rapid development of high-intensity lasers,high-sensitivity detectors and other optoelectronic devices,there have been reported many super-resolution imaging techniques that bypass the optical diffraction limit with different methods.Among these techniques,stimulated emission depletion microscopy(STED) technology has the advantages of high imaging resolution and fast imaging speed.This technology uses two lasers for imaging,one of which is used to excite fluorescence,and the other donut-shaped depletion laser is used to suppress the emission of fluorescent molecules around the fluorescent spot,in order to reduce the fluorescence point spread function and achieve super resolution Imaging.After recent years of development,the STED system has got great progress no matter from the generation,calibration and scanning of the beam,and the final imaging.In terms of laser source,new laser sources such as continuous wave beams,supercontinuum laser,stimulated Raman scattered laser,and higher-order Bessel beams have appeared;in scanning and calibration,new efficiency technology such as parallel scanning and automatic calibration have also appeared;In imaging,new methods such as time gating and phasor analysis have emerged to improve imaging quality.These new technologies and methods are of great significance to improve the efficiency of STED system construction and imaging.In addition,this paper also focuses on the ways to expand the imaging functions of the STED system.First,for three-dimensional STED imaging,this paper mainly introduces three methods to realize threedimensional STED imaging by wavefront non-coherent adjustment,4 Pi and structured light illumination methods.Second,for multi-color imaging,this paper introduces several dual-color and multi-color imaging techniques for special dyes.Third,this

关 键 词：受激辐射损耗显微 三维成像 多色成像 超分辨 

分 类 号：TP391.41[自动化与计算机技术—计算机应用技术] TH742[自动化与计算机技术—计算机科学与技术]