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Related Experiment Video

Updated: Jun 5, 2026

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

Published on: December 1, 2018

Probing GFP-actin diffusion in living cells using fluorescence correlation spectroscopy.

Hanna Engelke¹, Doris Heinrich, Joachim O Rädler

¹Center for NanoScience and Fakultät für Physik, Ludwig-Maximilians-Universität, Geschwister-Scholl-Platz 1, D-80539 München, Germany.

Physical Biology

|December 24, 2010

Summary

Fluorescence correlation spectroscopy (FCS) reveals dynamic changes in actin cytoskeleton. This technique measures the diffusion of green fluorescent protein-actin (GFP-actin) in living cells, providing insights into cytoskeletal properties.

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Determination of Lipid Raft Partitioning of Fluorescently-tagged Probes in Living Cells by Fluorescence Correlation Spectroscopy (FCS)

Determination of Lipid Raft Partitioning of Fluorescently-tagged Probes in Living Cells by Fluorescence Correlation Spectroscopy (FCS)

Published on: April 6, 2012

Area of Science:

Cell Biology
Biophysics

Background:

The eukaryotic cytoskeleton is dynamically remodeled by actin polymerization and depolymerization.
The balance between filamentous actin (F-actin) and globular actin (G-actin) fluctuates in space and time.
Understanding actin dynamics is crucial for cell structure and function.

Purpose of the Study:

To investigate the spatial and temporal signatures of the actin cytoskeleton using fluorescence correlation spectroscopy (FCS).
To determine the dynamics and indirectly the structural properties of cytoskeleton components with high spatial resolution.
To explore the diffusion of green fluorescent protein-labeled actin (GFP-actin) in living cells.

Main Methods:

Utilized fluorescence correlation spectroscopy (FCS) to measure the diffusion of GFP-actin in living Dictyostelium discoideum cells.
Employed free green fluorescent protein (GFP) as a reference for diffusion measurements.
Probed the FCS signal in the cortical F-actin network using the cytoskeleton protein LIM.

Main Results:

Actin diffusion within cells is primarily driven by G-actin and is slower than in diluted cell extracts.
Diffusion of G-actin in the cell cortex is slower than cytosolic G-actin diffusion.
Jasplakinolide-induced actin polymerization significantly reduced G-actin diffusion.
Latrunculin induced pronounced fluctuations in FCS correlation curves, indicative of actin waves.

Conclusions:

FCS of GFP-actin provides valuable information about local dynamics and cytoskeletal properties.
Combining FCS with scanning or spatial correlation techniques enhances the understanding of actin cytoskeleton dynamics.
This approach offers high spatial resolution for studying cytoskeletal remodeling.