Molecular orientation affects localization accuracy in superresolution far-field fluorescence microscopy

Nano Lett. 2011 Jan 12;11(1):209-13. doi: 10.1021/nl103472b. Epub 2010 Dec 6.

Abstract

We investigate the cooperative effect of molecular tilt and defocus on fluorophore localization by centroid calculation in far-field superresolution microscopy based on stochastic single molecule switching. If tilt angle and defocus are unknown, the localization contains systematic errors up to about ±125 nm. When imaging rotation-impaired fluorophores of unknown random orientation, the average localization accuracy in three-dimensional samples is typically limited to about ±32 nm, restricting the attainable resolution accordingly.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorescent Dyes / analysis*
  • Microscopy, Fluorescence / methods*
  • Sensitivity and Specificity
  • Stochastic Processes

Substances

  • Fluorescent Dyes