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Related Experiment Video

Updated: Jul 4, 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

New concepts for fluorescence correlation spectroscopy on membranes.

Jonas Ries¹, Petra Schwille

¹Biotechnologisches Zentrum, TU Dresden, Tatzberg 47-51, Dresden, D-01307, Germany.

Physical Chemistry Chemical Physics : PCCP

|June 13, 2008

Summary

Fluorescence correlation spectroscopy (FCS) offers insights into membrane properties but faces artifacts. Recent developments in FCS techniques enable accurate, quantitative measurements in complex membrane systems.

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Last Updated: Jul 4, 2026

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Area of Science:

Biophysics
Membrane Biophysics
Spectroscopy

Background:

Fluorescence correlation spectroscopy (FCS) is a valuable technique for analyzing molecular dynamics and concentrations in biological membranes.
Standard confocal FCS can be prone to artifacts, limiting its accuracy in complex or non-ideal membrane environments.

Purpose of the Study:

To evaluate the capabilities and limitations of confocal FCS for lipid membrane studies.
To present advanced FCS methodologies for overcoming artifacts and enabling quantitative measurements.

Main Methods:

Assessment of standard confocal FCS on lipid membranes.
Application of z-scan FCS and two-focus FCS for calibration-free measurements.
Utilizing scanning FCS for analyzing slow dynamics.
Employing surface-confined FCS to mitigate background noise.
Using variable detection area FCS to probe membrane heterogeneities.

Main Results:

Demonstrated calibration-free diffusion and concentration measurements using z-scan and two-focus FCS.
Presented methods for accurate measurement of slow dynamics with scanning FCS.
Showcased surface-confined FCS for studying membrane dynamics in challenging conditions.
Revealed submicroscopic heterogeneities in cell membranes via variable detection area FCS.

Conclusions:

Advanced FCS techniques significantly enhance the accuracy and quantitative capabilities for studying membrane systems.
These developments address limitations of standard FCS, enabling deeper insights into membrane biophysics and dynamics.