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Biophys. J. BioFAST: First Published September 30, 2005. doi:10.1529/biophysj.105.067959
© 2005 by the Biophysical Society.

A more recent version of this article appeared on December 1, 2005.
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Laure Wawrezinieck
Hervé Rigneault
Didier Marguet
Pierre-François Lenne
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MEMBRANES

Fluorescence Correlation Spectroscopy Diffusion Laws to Probe the Submicron Cell Membrane Organization

Laure Wawrezinieck 1, Hervé Rigneault 2, Didier Marguet 1 and Pierre-François Lenne 2*

1 Centre d'Immunologie de Marseille-Luminy
2 Institut Fresnel

* To whom correspondence should be addressed. E-mail: lenne{at}fresnel.fr.

Submitted on June 3, 2005
Revised on July 18, 2005
Accepted on 30 August 2005


  Abstract
To probe the complexity of the cell membrane organization and dynamics, it is important to obtain simple physical observables from experiments on live cells. Here we show that fluorescence correlation spectroscopy (FCS) measurements at different spatial scales enable to distinguish between different submicron confinement models. By plotting the diffusion time versus the transverse area of the confocal volume, we introduce the so-called 'FCS diffusion law', which is the key concept throughout this paper. First we report experimental FCS diffusion laws for two membrane constituents, which are respectively a putative raft marker and a cytoskeleton-hindered transmembrane protein. We find that these two constituents exhibit very distinct behaviors. To understand these results, we propose different models, which account for the diffusion of molecules either in a membrane comprising isolated microdomains or in a meshwork. By simulating FCS experiments for these two types of organization, we obtain FCS diffusion laws in agreement with our experimental observations. We also demonstrate that simple observables derived from these FCS diffusion laws are strongly related to confinement parameters such as the partition of molecules in microdomains and the average confinement time of molecules in a microdomain or a single mesh of a meshwork.

Key Words: anomalous diffusion, confinement time, cytoskeleton meshwork, hindered diffusion, lipid raft




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