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Molecular Diffusion in Plasma Membranes of Primary Lymphocytes Measured by Fluorescence Correlation Spectroscopy
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Total internal reflection fluorescence correlation spectroscopy: effects of lateral diffusion and surface-generated

Jonas Ries1, Eugene P Petrov, Petra Schwille

  • 1Biotechnologisches Zentrum, Technical University of Dresden, Dresden, Germany.

Biophysical Journal
|March 15, 2008
PubMed
Summary

This study enhances fluorescence correlation spectroscopy with total internal reflection excitation (TIR-FCS) by improving models to accurately measure molecular dynamics at surfaces. These advancements enable more precise quantitative analysis in biological interface studies.

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Area of Science:

  • Biophysics
  • Surface Science
  • Spectroscopy

Background:

  • Fluorescence correlation spectroscopy with total internal reflection excitation (TIR-FCS) is valuable for studying molecular interactions at surfaces.
  • Existing models for TIR-FCS correlation functions are often approximate and neglect lateral diffusion.

Purpose of the Study:

  • To develop accurate models for axial correlation functions in TIR-FCS.
  • To incorporate lateral diffusion and surface-generated emission into TIR-FCS models for improved accuracy.

Main Methods:

  • Extension of existing models for axial correlation functions.
  • Inclusion of lateral diffusion effects through realistic detection profiles.
  • Consideration of surface-generated emission in objective-based TIR-FCS.

Main Results:

  • Accurate expressions for correlation functions considering lateral diffusion.
  • Inclusion of surface-generated emission effects in objective-based TIR-FCS.
  • Development of models applicable to typical experimental conditions.

Conclusions:

  • The proposed models facilitate more quantitative and accurate measurements using TIR-FCS.
  • These advancements will improve the study of binding dynamics, diffusion coefficients, and concentrations at interfaces.
  • Enhanced TIR-FCS models offer greater precision for biological applications at planar substrates.