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Related Experiment Video

Updated: Jun 17, 2026

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

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 in living cells: a practical approach.

Nihal Altan-Bonnet¹, Grégoire Altan-Bonnet

¹Department of Biology, Rutgers University, Newark, New Jersey, USA.

Current Protocols in Cell Biology

|December 17, 2009

Summary

Fluorescence correlation spectroscopy (FCS) provides insights into molecular dynamics in living cells. This guide details live-cell FCS experiments using the Zeiss Confocor 3 for data acquisition and analysis.

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Area of Science:

Biophysics
Cell Biology
Microscopy

Background:

Fluorescence correlation spectroscopy (FCS) is a powerful single-molecule technique.
It enables the study of molecular dynamics within living cells.
FCS can quantify diffusion and molecular numbers of labeled proteins and lipids.

Purpose of the Study:

To provide a practical, step-by-step guide for performing live-cell FCS experiments.
To detail the setup, data acquisition, and analysis procedures.
To demonstrate the application of FCS using the Zeiss Confocor 3 system.

Main Methods:

Utilizing a commercial spectroscopy/microscopy system (Zeiss Confocor 3).
Establishing protocols for live-cell FCS experiments.
Implementing reliable data acquisition and analysis techniques.

Main Results:

Demonstration of a feasible workflow for live-cell FCS.
Successful acquisition and analysis of molecular dynamics data.
Quantification of diffusion timescales and molecular counts in cellular environments.

Conclusions:

This unit offers a comprehensive protocol for live-cell FCS.
It empowers researchers to investigate molecular dynamics in biological systems.
The described methods facilitate robust data generation and interpretation.