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Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

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Fluorescence correlation spectroscopy: past, present, future.

Elliot L Elson¹

¹Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, Washington University in St. Louis, St. Louis, Missouri, USA. elson@wustl.edu

Biophysical Journal

|January 3, 2012

Summary

Fluorescence Correlation Spectroscopy (FCS) is a powerful technique for measuring molecular properties in small volumes like cells. It bridges ensemble and single-molecule methods, offering insights into dynamic systems and fluctuations.

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Area of Science:

Biophysics
Chemical Physics
Physical Chemistry

Background:

Fluorescence Correlation Spectroscopy (FCS) is a routine method for quantifying molecular parameters.
FCS bridges classical ensemble and single-molecule measurements by analyzing temporal and spatial correlations.
It provides insights into molecular concentrations and fluctuations in nonlinear systems.

Purpose of the Study:

To highlight the utility of FCS in determining various molecular parameters.
To emphasize FCS's role in characterizing dynamic systems, particularly biological cells.
To explore FCS's potential in studying nonequilibrium steady states in living systems.

Main Methods:

FCS utilizes fluorescence microscopy to analyze molecular behavior.
It measures spatial and temporal correlations of individual molecules.
The technique samples femtoliter volumes, ideal for small dynamic systems.

Main Results:

FCS accurately determines diffusion coefficients, rate constants, concentrations, and fluorescence brightness.
It provides information on triplet state lifetimes and molecular number fluctuations.
FCS offers a unique perspective on mesoscopic systems and fluctuations.

Conclusions:

FCS is a versatile tool for molecular characterization in biophysical and chemical research.
Its application extends to understanding complex dynamics in biological cells.
FCS opens new avenues for studying nonequilibrium steady states, especially in living cells.