Superresolution imaging using single-molecule localization

Annu Rev Phys Chem. 2010;61:345-67. doi: 10.1146/annurev.physchem.012809.103444.

Abstract

Superresolution imaging is a rapidly emerging new field of microscopy that dramatically improves the spatial resolution of light microscopy by over an order of magnitude (approximately 10-20-nm resolution), allowing biological processes to be described at the molecular scale. Here, we discuss a form of superresolution microscopy based on the controlled activation and sampling of sparse subsets of photoconvertible fluorescent molecules. In this single-molecule-based imaging approach, a wide variety of probes have proved valuable, ranging from genetically encodable photoactivatable fluorescent proteins to photoswitchable cyanine dyes. These have been used in diverse applications of superresolution imaging: from three-dimensional, multicolor molecule localization to tracking of nanometric structures and molecules in living cells. Single-molecule-based superresolution imaging thus offers exciting possibilities for obtaining molecular-scale information on biological events occurring at variable timescales.

Publication types

  • Review

MeSH terms

  • Image Interpretation, Computer-Assisted
  • Luminescent Proteins / chemistry
  • Microscopy, Fluorescence / methods*
  • Microscopy, Polarization / methods

Substances

  • Luminescent Proteins