Total internal reflection fluorescence correlation spectroscopy: Effects of lateral diffusion and surface-generated fluorescence

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Jonas Ries - , TUD Dresden University of Technology (Author)
  • Eugene P. Petrov - , TUD Dresden University of Technology (Author)
  • Petra Schwille - , Chair of Biophysics (Author)

Abstract

Fluorescence correlation spectroscopy with total internal reflection excitation (TIR-FCS) is a promising method with emerging biological applications for measuring binding dynamics of fluorescent molecules to a planar substrate as well as diffusion coefficients and concentrations at the interface. Models for correlation functions proposed so far are rather approximate for most conditions, since they neglect lateral diffusion of fluorophores. Here we propose accurate extensions of previously published models for axial correlation functions taking into account lateral diffusion through detection profiles realized in typical experiments. In addition, we consider the effects of surface-generated emission in objective-based TIR-FCS. The expressions for correlation functions presented here will facilitate quantitative and accurate measurements with TIR-FCS.

Details

Original languageEnglish
Pages (from-to)390-399
Number of pages10
JournalBiophysical journal
Volume95
Issue number1
Publication statusPublished - 1 Jul 2008
Peer-reviewedYes

External IDs

PubMed 18339763

Keywords

ASJC Scopus subject areas