Multicolor super-resolution imaging with photo-switchable fluorescent probes

Science. 2007 Sep 21;317(5845):1749-53. doi: 10.1126/science.1146598. Epub 2007 Aug 16.

Abstract

Recent advances in far-field optical nanoscopy have enabled fluorescence imaging with a spatial resolution of 20 to 50 nanometers. Multicolor super-resolution imaging, however, remains a challenging task. Here, we introduce a family of photo-switchable fluorescent probes and demonstrate multicolor stochastic optical reconstruction microscopy (STORM). Each probe consists of a photo-switchable "reporter" fluorophore that can be cycled between fluorescent and dark states, and an "activator" that facilitates photo-activation of the reporter. Combinatorial pairing of reporters and activators allows the creation of probes with many distinct colors. Iterative, color-specific activation of sparse subsets of these probes allows their localization with nanometer accuracy, enabling the construction of a super-resolution STORM image. Using this approach, we demonstrate multicolor imaging of DNA model samples and mammalian cells with 20- to 30-nanometer resolution. This technique will facilitate direct visualization of molecular interactions at the nanometer scale.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Chlorocebus aethiops
  • Clathrin-Coated Vesicles
  • DNA / analysis*
  • DNA Probes*
  • Fluorescent Dyes*
  • Microscopy, Fluorescence / methods
  • Microtubules
  • Nanotechnology

Substances

  • DNA Probes
  • Fluorescent Dyes
  • DNA