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Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Updated: Feb 16, 2026

Spot Variation Fluorescence Correlation Spectroscopy for Analysis of Molecular Diffusion at the Plasma Membrane of Living Cells

Spot Variation Fluorescence Correlation Spectroscopy for Analysis of Molecular Diffusion at the Plasma Membrane of Living Cells

Published on: November 12, 2020

Circle scanning STED fluorescence correlation spectroscopy to quantify membrane dynamics and compartmentalization.

Riccardo Maraspini¹, Oliver Beutel¹, Alf Honigmann¹

¹Max Planck Institute of Cell Biology and Genetics, 01309 Dresden, Germany.

Methods (San Diego, Calif.)

|December 21, 2017

Summary

This study introduces Circle Scanning Stimulated Emission Depletion Fluorescence Correlation Spectroscopy (CS-STED-FCS) for precise measurement of molecular membrane dynamics. The technique maps cellular membrane heterogeneities with high spatiotemporal resolution.

Keywords:

Lipid phase separation Membrane dynamics Scanning fluorescence correlation spectroscopy Sub-diffusion Super-resolution STED microscopy

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Area of Science:

Biophysics
Cell Biology
Microscopy

Background:

Understanding cell membrane organization and function requires quantifying molecular dynamics.
Complex membrane structures necessitate high spatial and temporal resolution for accurate interpretation.

Purpose of the Study:

To describe the Circle Scanning Fluorescence Correlation Spectroscopy combined with Stimulated Emission Depletion Microscopy (CS-STED-FCS) method.
To enable quantification of sub-diffusion processes and mapping of membrane heterogeneities.

Main Methods:

Utilizing Circle Scanning Fluorescence Correlation Spectroscopy (CS-FCS).
Integrating CS-FCS with Stimulated Emission Depletion (STED) microscopy.
Calibrating and testing the technique with model membranes.
Applying the method to living cells.

Main Results:

CS-STED-FCS allows quantification of sub-diffusion processes.
The method provides direct mapping of membrane heterogeneities.
High spatiotemporal resolution and good statistics were achieved in living cells.

Conclusions:

CS-STED-FCS is a powerful technique for high-resolution membrane dynamics studies.
The method offers precise insights into cell membrane organization and function.
This technique advances the study of molecular dynamics in biological membranes.