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Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.

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Studying flow close to an interface by total internal reflection fluorescence cross-correlation spectroscopy:

R Schmitz1, S Yordanov, H J Butt

  • 1Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany.

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|February 7, 2012
PubMed
Summary
This summary is machine-generated.

This study introduces a quantitative method combining Brownian dynamics simulations and experiments to precisely measure hydrodynamic flow properties near surfaces using total internal reflection fluorescence cross-correlation spectroscopy (TIR-FCCS). The method accurately determines shear rate and slip length, revealing near-zero slip on hydrophilic surfaces.

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Area of Science:

  • Surface science
  • Fluid dynamics
  • Spectroscopy

Background:

  • Total internal reflection fluorescence cross-correlation spectroscopy (TIR-FCCS) probes nanoscale hydrodynamic flows near surfaces.
  • Current TIR-FCCS methods provide indirect flow information, lacking quantitative accuracy for parameters like shear rate and slip length.

Purpose of the Study:

  • To develop a quantitative method for retrieving hydrodynamic flow properties from TIR-FCCS measurements.
  • To combine theoretical modeling with experimental data for accurate analysis.
  • To determine the slip length of a fluid near a hydrophilic surface.

Main Methods:

  • Utilizing Brownian dynamics simulations to generate accurate correlation functions for model parameters.
  • Employing an importance-sampling Monte Carlo procedure to fit experimental correlation functions.
  • Systematically varying simulation parameters to determine optimal values and statistical error bars.

Main Results:

  • A quantitative method was established to retrieve flow properties from TIR-FCCS data.
  • The approach is computationally efficient, suitable for parallel processing.
  • Flow near a hydrophilic surface exhibited a slip length below 10nm, indistinguishable from zero within experimental and model limitations.

Conclusions:

  • The combined simulation and experimental approach provides accurate quantitative analysis of hydrodynamic flows near surfaces.
  • The method successfully determined a near-zero slip length for flow past a hydrophilic surface.
  • This technique enhances the utility of TIR-FCCS for surface-dependent fluid dynamics studies.