Journal of Fluorescence

, Volume 20, Issue 1, pp 105–114

On the Resolution Capabilities and Limits of Fluorescence Lifetime Correlation Spectroscopy (FLCS) Measurements

  • Steffen Rüttinger
  • Peter Kapusta
  • Matthias Patting
  • Michael Wahl
  • Rainer Macdonald
Original Paper

Abstract

Quantitative tests were performed in order to explore the practical limits of FLCS. We demonstrate that: a) FLCS yields precise and correct concentration values from as low as picomolar to micromolar concentrations; b) it is possible to separate four signal components in a single detector setup; c) diffusion times differing only 25% from each other can be resolved by separating a two component mixture based on the different fluorescence lifetimes of both components; d) most of the inherent technical limitations of conventional FCS are easily overcome by FLCS employing a single detector channel confocal detection scheme.

Keywords

Fluorescence lifetime correlation spectroscopy (FLCS) Fluorescence correlation spectroscopy (FCS) Single molecule detection (SMD) Confocal laser scanning microscopy (CLSM) 

Supplementary material

10895_2009_528_MOESM1_ESM.doc (34 kb)
Supplemental Material 1(DOC 33.5 kb)

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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Steffen Rüttinger
    • 1
  • Peter Kapusta
    • 2
  • Matthias Patting
    • 2
  • Michael Wahl
    • 2
  • Rainer Macdonald
    • 1
  1. 1.Physikalisch-Technische BundesanstaltBerlinGermany
  2. 2.PicoQuant GmbHBerlinGermany

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