Fluorescence correlation spectroscopy in semiadhesive wall proximity

Anal Chem. 2011 Nov 1;83(21):8101-7. doi: 10.1021/ac201116g. Epub 2011 Oct 4.

Abstract

With examination of diffusion in heterogeneous media through fluorescence correlation spectroscopy, the temporal correlation of the intensity signal shows a long correlation tail and the characteristic diffusion time results are no longer easy to determine. Excluded volume and sticking effects have been proposed to justify such deviations from the standard behavior since all contribute and lead to anomalous diffusion mechanisms . Usually, the anomalous coefficient embodies all the effects of environmental heterogeneity providing too general explanations for the exotic diffusion recorded. Here, we investigated whether the reason of anomalies could be related to a lack of an adequate interpretative model for heterogeneous systems and how the presence of obstacles on the detection volume length scale could affect fluorescence correlation spectroscopy experiments. We report an original modeling of the autocorrelation function where fluorophores experience reflection or adsorption at a wall placed at distances comparable with the detection volume size. We successfully discriminate between steric and adhesion effects through the analysis of long time correlations and evaluate the adhesion strength through the evaluation of probability of being adsorbed and persistence time at the wall on reference data. The proposed model can be readily adopted to gain a better understanding of intracellular and nanoconfined diffusion opening the way for a more rational analysis of the diffusion mechanism in heterogeneous systems and further developing biological and biomedical applications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Adhesion*
  • Cell Membrane*
  • Diffusion
  • Fluorescent Dyes / chemistry*
  • Humans
  • Models, Biological*
  • Nanotubes*
  • Spectrometry, Fluorescence*

Substances

  • Fluorescent Dyes