Laser & Optoelectronics Progress, Volume. 57, Issue 24, (2020)

Review and Prospect for Single Molecule Localization Microscopy

Yuzhu Li1, Chuankang Li1, Xiang Hao1, Xu Liu1,3, and Cuifang Kuang1,2,3、*
Author Affiliations
  • 1State Key Laboratory of Modern Optical Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China
  • 2Ningbo Institute, Zhejiang University, Ningbo, Zhejiang 315100, China
  • 3Collaborative Innovation Center of Extreme Optics, Shanxi University, Taiyuan, Shanxi 030006, China
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    Figures & Tables(8)
    Research history of single molecule localization microscopy
    Principle of fluorescence emission
    Basic configuration of fluorescence microscopy
    Schematic of three-dimensional STORM[20]. (a) Experimental setup; (b) relationship between axial lengths of elliptical PSF and z position of molecules
    Principle diagram of stochastic photobleaching super-resolution imaging [10]. (a) Attenuation of integrated intensity with time; (b) distinguishing two PSFs by photobleaching
    Setup and principle diagram of MINFLUX[29]. (a) Experimental setup of MINFLUX; (b) detection steps of MINFLUX
    Principle diagram of SIMFLUX[34]. (a) Experimental setup; (b) work flow chart
    • Table 1. Comparison among all SMLMs

      View table

      Table 1. Comparison among all SMLMs

      SMLMBest resolutionLocalizationprecisionAdvantageDisadvantage
      PALM [16]2-25nmσ/NSuper high precisionBeing time-consuming, large amount of calculation, being unsuitable for dynamic process imaging, severe photobleaching, and only exogenous proteins being observed
      STORM[17]About 20nmσ/NSuper high precisionBeing limited in in-vivo imaging, severe photobleaching, and being unable to quantify molecule numbers in a cell
      PAINT[18]About 25nmσ/NNo need to label sampleLarge amount of calculation, long post-processing time, and only obtaining surface information of sample, not internal information
      GSDIM[22]About 30nmσ/NNo need of special fluorophore, simple device, and shortening time of image acquisitionSevere photobleaching, complex processing, and being limited in real-time imaging of living cells
      SHRImP [10]About 10nmσ/NNo special fluorophore, and relatively simple processBeing unsuitable for cases where two or more neighboring molecules are bleached at the same time and being limited in 2D situation
      SRRF[28]50-106nmσ/NBeing able to be complemented in conventional microscopeBeing limited resolution
      SOFI[25]100nmσ/NNo phototoxicity and short processing timeBeing limited resolution
      MINFLUX[29]1-5nmAbout L/22NImproved photon utilization, improved localization precision, and being suitable for living cell imagingComplex processing and being limited in sparse solution and long acquisition time
      SIMFLUX[34]5-10nmσ/2.4NImproved localization precision and accuracyBeing limited in 2D situation
      ROSE[33]2-5nmAbout 50/NImproved localization precision and accuracyBeing time-consuming
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    Yuzhu Li, Chuankang Li, Xiang Hao, Xu Liu, Cuifang Kuang. Review and Prospect for Single Molecule Localization Microscopy[J]. Laser & Optoelectronics Progress, 2020, 57(24):

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    Paper Information

    Category: Reviews

    Received: Apr. 7, 2020

    Accepted: May. 8, 2020

    Published Online: Dec. 30, 2020

    The Author Email: Kuang Cuifang (cfkuang@zju.edu.cn)

    DOI:10.3788/LOP57.240002

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