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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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Modeling Active Particle Motion from Fluorescence Correlation Spectroscopy Data.

Miguel A Ramos-Docampo¹, Cairui Duan², Nanying Wang¹

¹Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000 Aarhus, Denmark.

Analytical Chemistry

|June 9, 2026

Summary

This summary is machine-generated.

Fluorescence correlation spectroscopy (FCS) in 3D complements 2D optical tracking for analyzing active colloids. Both methods reveal consistent motor dynamics, aiding transport property assessment.

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Area of Science:

Colloid and Interface Science
Soft Matter Physics
Nanotechnology

Background:

Active colloids are engineered micro/nanomotors with diverse propulsion mechanisms.
Comparing 2D and 3D analysis techniques for active colloids is crucial but challenging.
Fluorescence correlation spectroscopy (FCS) offers a 3D volumetric approach.

Purpose of the Study:

To compare 3D FCS measurements with 2D optical tracking for active colloids.
To establish FCS detectability criteria for active colloids.
To quantify locomotion and distinguish between active and passive colloids.

Main Methods:

Utilized fluorescence correlation spectroscopy (FCS) for 3D particle motion analysis.
Employed standard 2D optical microscopy-based tracking.
Investigated three propulsion types: self-diffusiophoresis, bubble propulsion, and magnetic guidance.

Main Results:

FCS successfully quantified locomotion in single and mixed active/passive colloid populations.
Directed particle motion assessed by FCS yielded velocities comparable to optical tracking.
Consistent trends in motor dynamics were observed, though absolute diffusion coefficients differed.

Conclusions:

FCS is a reliable method for assessing active colloid transport properties with appropriate data treatment.
FCS and optical tracking are complementary, providing population-level and individual motor diffusivity, respectively.
This study bridges the gap between 2D and 3D analysis methods for active matter.