AI Article Synopsis
- Understanding the dynamic interactions between proteins and lipids in cell membranes is a key focus in cell biology, but current methods for observing these interactions in real-time are lacking.
- Recent advancements in spot-variation fluorescence correlation spectroscopy allow scientists to measure the transient confinement times of membrane components with much greater precision than before.
- This technique, which uses standard optics, helps to analyze specific binding mechanisms and provides recommendations for experimental setups to better understand how membrane constituents interact on a molecular level.
Article 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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3117160 | PMC |
| http://dx.doi.org/10.1016/j.bpj.2011.04.035 | DOI Listing |
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