Ultrahigh-resolution colocalization of spectrally separable point-like fluorescent probes

Methods. 2001 Sep;25(1):87-102. doi: 10.1006/meth.2001.1218.

Abstract

An ultrahigh-resolution colocalization method based on the simultaneous acquisition and analysis of spectrally separated images of the excitation point-spread function of point-like fluorescent probes is reviewed. It is shown that molecular distances can be measured with accuracy better than 10 nm using conventional far-field optics. A detailed account of the methodology, theoretical considerations, signal processing, and data fitting algorithms is given.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.
  • Review

MeSH terms

  • Fluorescent Dyes / isolation & purification*
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Photons
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence / instrumentation
  • Spectrometry, Fluorescence / methods*
  • Transducers

Substances

  • Fluorescent Dyes