超分辨显微成像技术在活细胞成像中的应用与发展

Application and Development of Super-resolution Microscopy in Live Cell Imaging

超分辨显微成像技术(super-resolution microscopy,SRM)可以绕过光学衍射极限对成像分辨率的限制,让以前观察不到的纳米级结构实现可视化,这一重大研究进展推动了现代生命科学和生物医学研究的进步与发展.细胞是生物体的基本组成单位,对活细胞内部的细微结构和动力学过程进行研究是掌握生命本质必不可少的途径.但由于成像原理或条件的限制,早期的SRM技术在活细胞成像应用方面受到了不同程度的限制.近几年来,随着SRM和相关技术的发展,SRM在活细胞成像研究中的应用也越来越多.本文简要介绍目前常见的几种SRM技术的基本原理和特点,并在此基础上着重阐述它们在活细胞成像应用中所取得的最新研究进展和发展方向.

Super-resolution microscopy(SRM)can bypass the optical diffraction limitation to imaging resolution and visualize previously unobservable nanoscale structures.This significant breakthrough has promoted the progress and development of modern life science and biomedical research.Cell is the basic unit of living organisms,and studying the fine structures and dynamic processes inside living cells is an indispensable way to grasp the essence of life.However,due to the limitations of imaging principles or conditions,the applications of early SRM technologies in live cell imaging were limited in different degrees.In recent years,with the development of SRM technology and related technologies,the application of SRM in live cell imaging research has also been increasingly developed.This paper briefly introduces the basic principles and characteristics of several common SRM technologies,and on this basis,focuses on the latest research progress and development trend in their live-cell-imaging applications.For STORM,the developments of high-density localization algorithms and new blinking fluorescent probes promote its application in live cell super-resolution imaging,realizing the monitoring of the distribution and changes of specific proteins in the nucleus of living cells,as well as the super-resolution imaging of the dynamic process of organelle membranes in living cells.For STED,the developments of new fluorescent probes with high photostability,good bleaching resistance and low saturation threshold,and the utilization of new methods such as time-gated technology for reducing the depletion power help with its application in live cell imaging.The long-term super-resolution monitoring of the dynamic process of specific organelles in living cells and the super-resolution imaging of the interactions between different organelles in living cells are realized.For SIM,by introducing ingenious methods such as TIRF illumination,pattern activation and grazing incidence,researchers try to improve its resolution while retaining its advantage of live cell imaging,and realize the dynamic super-resolution imaging within 100 nm scale and with fast imaging speed in living cells.