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Issue 24, 2008
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New concepts for fluorescence correlation spectroscopy on membranes

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Abstract

Fluorescence correlation spectroscopy (FCS) is a powerful tool to measure useful physical quantities such as concentrations, diffusion coefficients, diffusion modes or binding parameters, both in model and cell membranes. However, it can suffer from severe artifacts, especially in non-ideal systems. Here we assess the potential and limitations of standard confocal FCS on lipid membranes and present recent developments which facilitate accurate and quantitative measurements on such systems. In particular, we discuss calibration-free diffusion and concentration measurements using z-scan FCS and two focus FCS and present several approaches using scanning FCS to accurately measure slow dynamics. We also show how surface confined FCS enables the study of membrane dynamics even in presence of a strong cytosolic background and how FCS with a variable detection area can reveal submicroscopic heterogeneities in cell membranes.

Graphical abstract: New concepts for fluorescence correlation spectroscopy on membranes

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Article information


Submitted
23 Nov 2007
Accepted
03 Mar 2008
First published
27 Mar 2008

Phys. Chem. Chem. Phys., 2008,10, 3487-3497
Article type
Perspective

New concepts for fluorescence correlation spectroscopy on membranes

J. Ries and P. Schwille, Phys. Chem. Chem. Phys., 2008, 10, 3487
DOI: 10.1039/B718132A

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