Anomalous transport resolved in space and time by fluorescence correlation spectroscopy

TL;DR

This paper develops a theoretical framework for space-resolved fluorescence correlation spectroscopy (FCS) to distinguish different microscopic mechanisms of subdiffusion in crowded cell membranes, validated by simulations and compatible with experimental data.

Contribution

It generalizes FCS theory beyond Gaussian transport assumptions and introduces a master formula for the autocorrelation function to identify subdiffusion mechanisms.

Findings

FCS can differentiate between fractional Brownian motion and hindered diffusion.

Derived a master formula linking FCS autocorrelation to microscopic transport mechanisms.

Validated scaling predictions with in silico experiments and experimental data.

Abstract

A ubiquitous observation in crowded cell membranes is that molecular transport does not follow Fickian diffusion but exhibits subdiffusion. The microscopic origin of such a behaviour is not understood and highly debated. Here we discuss the spatio-temporal dynamics for two models of subdiffusion: fractional Brownian motion and hindered motion due to immobile obstacles. We show that the different microscopic mechanisms can be distinguished using fluorescence correlation spectroscopy (FCS) by systematic variation of the confocal detection area. We provide a theoretical framework for space-resolved FCS by generalising FCS theory beyond the common assumption of spatially Gaussian transport. We derive a master formula for the FCS autocorrelation function, from which it is evident that the beam waist of an FCS experiment is a similarly important parameter as the wavenumber of scattering experiments. These results lead to scaling properties of the FCS correlation for both models, which are tested by in silico experiments. Further, our scaling prediction is compatible with the FCS half-value times reported by Wawrezinieck et al. [Biophys. J. 89, 4029 (2005)] for in vivo experiments on a transmembrane protein.