Spot variation fluorescence correlation spectroscopy allows for superresolution chronoscopy of confinement times in membranes.

Verena Ruprecht (1) , Stefan Wieser (2) , Didier Marguet (2) , Gerhard J Schütz (1)

1 JKU - University of Linz - Johannes Kepler Universität Linz
2 CIML - Centre d'Immunologie de Marseille - Luminy

Resolving the dynamical interplay of proteins and lipids in the live-cell plasma membrane represents a central goal in current cell biology. Superresolution concepts have introduced a means of capturing spatial heterogeneity at a nanoscopic length scale. Similar concepts for detecting dynamical transitions (superresolution chronoscopy) are still lacking. Here, we show that recently introduced spot-variation fluorescence correlation spectroscopy allows for sensing transient confinement times of membrane constituents at dramatically improved resolution. Using standard diffraction-limited optics, spot-variation fluorescence correlation spectroscopy captures signatures of single retardation events far below the transit time of the tracer through the focal spot. We provide an analytical description of special cases of transient binding of a tracer to pointlike traps, or association of a tracer with nanodomains. The influence of trap mobility and the underlying binding kinetics are quantified. Experimental approaches are suggested that allow for gaining quantitative mechanistic insights into the interaction processes of membrane constituents.

Domaines: Immunologie

Verena Ruprecht, Stefan Wieser, Didier Marguet, Gerhard J Schütz. Spot variation fluorescence correlation spectroscopy allows for superresolution chronoscopy of confinement times in membranes.. Biophysical Journal, 2011, 100 (11), pp.2839-45. ⟨10.1016/j.bpj.2011.04.035⟩. ⟨hal-00611626⟩