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Related Experiment Videos

Total internal reflection with fluorescence correlation spectroscopy: nonfluorescent competitors.

Alena M Lieto1, Nancy L Thompson

  • 1Department of Physics and Astronomy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.

Biophysical Journal
|August 10, 2004
PubMed
Summary
This summary is machine-generated.

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Total internal reflection fluorescence correlation spectroscopy quantifies molecular interactions at surfaces. This study details how competing fluorescent and nonfluorescent molecules affect surface binding kinetics and concentrations.

Area of Science:

  • Biophysics
  • Surface Science
  • Analytical Chemistry

Background:

  • Total internal reflection fluorescence correlation spectroscopy (TIR-FCS) measures molecular dynamics at interfaces.
  • Existing models describe diffusion and surface kinetics but not competitive binding.

Purpose of the Study:

  • To describe the autocorrelation function in TIR-FCS when both surface kinetics and diffusion are present.
  • To analyze competitive binding between fluorescent and nonfluorescent molecules for surface sites.

Main Methods:

  • Theoretical analysis of the fluorescence fluctuation autocorrelation function.
  • Modeling the influence of competing molecules on surface binding.

Main Results:

  • The autocorrelation function is influenced by association/dissociation rate constants for both fluorescent and nonfluorescent molecules.

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  • Key parameters include surface site density, solution concentrations, diffusion coefficients, evanescent field depth, and observation area.

    • Conclusions:

    • This work extends TIR-FCS analysis to include competitive binding scenarios.
    • Provides a theoretical framework for more complex interfacial molecular interaction studies.