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Updated: Jul 10, 2026

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
Published on: December 11, 2021
Fluorescence correlation spectroscopy and its application to the characterization of molecular properties and
Hacène Boukari1, Dan L Sackett
1Laboratory of Integrative and Medical Biophysics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA.
Abstract:
Fluorescence correlation spectroscopy (FCS) utilizes temporal fluctuations in fluorescence emission to extract quantitative measures of inter- or intramolecular dynamics or molecular motions of probe molecules, which occur on submicrosecond to second timescales. In typical experiments, one can readily obtain the probe's diffusion coefficient and concentration from small volumes of sample. Recent FCS applications have yielded information on interactions of the probe with changing or structured solvent, binding with other molecules, photophysical or conformational changes in the probe, polymerization, and other changes in the dynamics of the probe. In cross-correlation mode FCS promises to attract more applications as the technique can monitor interactions in a system with two or more probes with different fluorophores.
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