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It has recently been proposed, with the help of numerical investigations, that fluorescence correlation spectroscopy at variable observation area can reveal the existence of a meshgrid of semi-permeable barriers hindering the two-dimensional diffusion of tagged particles, such as plasmic membrane constituents. We present a complete theory confirming and accounting for these findings. It enables a reliable, quantitative exploitation of experimental data from which the sub-wavelength mesh size can be extracted. Time-scales at which fluorescence correlation spectroscopy must be performed experimentally are discussed in detail.

Graphical abstract: Theory of fluorescence correlation spectroscopy at variable observation area for two-dimensional diffusion on a meshgrid

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