Application and Development of Super-resolution Microscopy in Live Cell Imaging
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Center for Biomedical Photonics & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems, Shenzhen University, Shenzhen 518060, China

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This work was supported by grants from The National Natural Science Foundation of China (61775144, 61975131, 61620106016, 61525503, 61835009), the Natural Science Foundation of Guangdong Province (2018A030313362) and the Shenzhen Research Project (JCYJ20170818144012025, JCYJ20200109105411133, JCYJ20170818141701667, JCYJ20170412105003520).

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    Abstract:

    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.

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ZHANG Jiao, HE Qin, WU Ze-Kai, YU Bin, QU Jun-Le, LIN Dan-Ying. Application and Development of Super-resolution Microscopy in Live Cell Imaging[J]. Progress in Biochemistry and Biophysics,2021,48(11):1301-1315

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History
  • Received:December 05,2020
  • Revised:March 03,2021
  • Accepted:April 26,2021
  • Online: November 23,2021
  • Published: November 20,2021