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

Abstract:

: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.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Ruprecht V,Wieser S,Marguet D,Schütz GJ

doi

10.1016/j.bpj.2011.04.035

subject

Has Abstract

pub_date

2011-06-08 00:00:00

pages

2839-45

issue

11

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(11)00481-4

journal_volume

100

pub_type

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