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Related Experiment Videos

lambda-Repressor oligomerization kinetics at high concentrations using fluorescence correlation spectroscopy in

K T Samiee1, M Foquet, L Guo

  • 1School of Applied & Engineering Physics, Cornell University, Ithaca, New York 14853, USA.

Biophysical Journal
|December 23, 2004
PubMed
Summary
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Zero-mode waveguides reduce the observation volume for fluorescence correlation spectroscopy (FCS), enabling high-concentration measurements. This technique successfully measured protein oligomerization and biological activity.

Area of Science:

  • Biophysics
  • Optical Nanostructures
  • Spectroscopy

Background:

  • Fluorescence correlation spectroscopy (FCS) is effective for kinetics and transport at low concentrations.
  • High fluorophore concentrations limit traditional FCS applications.

Purpose of the Study:

  • To adapt FCS for higher fluorophore concentrations using zero-mode waveguides.
  • To measure protein oligomerization and biological activity with enhanced FCS.

Main Methods:

  • Utilized zero-mode waveguides to reduce the FCS observation volume.
  • Derived an empirical FCS model for 1D diffusion in a finite tube.
  • Applied the technique to bacteriophage lambda repressor protein.

Main Results:

Related Experiment Videos

  • Successfully applied FCS to micromolar protein concentrations.
  • Achieved results consistent with previous studies.
  • Demonstrated assaying biological activity via diffusion constant changes.

    • Conclusions:

    • Zero-mode waveguides significantly enhance FCS capabilities for higher concentrations.
    • This method is valuable for studying protein dynamics and ligand binding.
    • The technique offers a new approach for biological activity assays.