Mean squared displacement from fluorescence correlation spectroscopy
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Updated: Mar 24, 2026

From Fast Fluorescence Imaging to Molecular Diffusion Law on Live Cell Membranes in a Commercial Microscope
Published on: October 9, 2014
Jakub Kubečka1, Filip Uhlík1, Peter Košovan1
1Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University in Prague, Hlavova 8, 128 00 Praha 2, Czech Republic. peter.kosovan@natur.cuni.cz.
Abstract:
Under certain conditions, the mean squared displacement (MSD) can be retrieved from fluorescence correlation spectroscopy (FCS) measurements. However, in the general case this procedure is not valid, and the apparent MSD obtained from FCS data may substantially differ from the true one. In this work we discuss under which conditions this procedure can be applied. Furthermore, we use computer simulations to obtain the MSD and the apparent MSD for the diffusion of a single polymer chain under various approximations. Based on the simulation results we discuss the reliability of the apparent MSD obtained from FCS, showing that it systematically deviates from the true MSD. We also propose a general procedure to verify the reliability of the apparent MSD by measurements at various focal spot sizes.
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