Lucky imaging: improved localization accuracy for single molecule imaging

Biophys J. 2009 Apr 8;96(7):2912-7. doi: 10.1016/j.bpj.2008.12.3945.


We apply the astronomical data-analysis technique, Lucky imaging, to improve resolution in single molecule fluorescence microscopy. We show that by selectively discarding data points from individual single-molecule trajectories, imaging resolution can be improved by a factor of 1.6 for individual fluorophores and up to 5.6 for more complex images. The method is illustrated using images of fluorescent dye molecules and quantum dots, and the in vivo imaging of fluorescently labeled linker for activation of T cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / analysis
  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • Cell Line, Tumor
  • Cell Survival
  • Fluorescence
  • Fluorescent Dyes / metabolism
  • Image Processing, Computer-Assisted
  • Membrane Proteins / analysis
  • Membrane Proteins / metabolism
  • Mice
  • Microscopy, Fluorescence / methods*
  • Phosphoproteins / analysis
  • Phosphoproteins / metabolism
  • Quantum Dots
  • Sensitivity and Specificity
  • T-Lymphocytes / cytology
  • T-Lymphocytes / metabolism


  • Adaptor Proteins, Signal Transducing
  • Fluorescent Dyes
  • Lat protein, mouse
  • Membrane Proteins
  • Phosphoproteins