ArticleBiophysical journal2003
Ligand-receptor kinetics measured by total internal reflection with fluorescence correlation spectroscopy.
Article in Biophysical journal, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed.
- Mapping Protein Diffusion from the Plasma Membrane to the Nucleus: Insights from Fluorescence Correlation Spectroscopy.Biomolecules · 2026Review
- Photo-Induced Depletion of Binding Sites in DNA-PAINT Microscopy.Molecules (Basel, Switzerland) · 2018Article
- Surface plasmon-assisted microscope.Journal of biomedical optics · 2018Article
- Quantifying Reversible Surface Binding via Surface-Integrated Fluorescence Correlation Spectroscopy.Nano letters · 2018Article
- Insights into Protein-Ligand Interactions: Mechanisms, Models, and Methods.International journal of molecular sciences · 2016Review
- Direct observation of T4 lysozyme hinge-bending motion by fluorescence correlation spectroscopy.Biophysical journal · 2012Article
- Determination of lipid raft partitioning of fluorescently-tagged probes in living cells by Fluorescence Correlation Spectroscopy (FCS).Journal of visualized experiments : JoVE · 2012Article
- Fluorescence correlation spectroscopy: past, present, future.Biophysical journal · 2011Review
- Measuring surface binding thermodynamics and kinetics by using total internal reflection with fluorescence correlation spectroscopy: practical considerations.The journal of physical chemistry. B · 2011Article
- Imaging with total internal reflection fluorescence microscopy for the cell biologist.Journal of cell science · 2010Review
- Kinetics of M1 muscarinic receptor and G protein signaling to phospholipase C in living cells.The Journal of general physiology · 2010Article
- Total internal reflection with fluorescence correlation spectroscopy: Applications to substrate-supported planar membranes.Journal of structural biology · 2009Article
- Fluorescence fluctuation spectroscopy: ushering in a new age of enlightenment for cellular dynamics.Biophysical reviews · 2009Article
- Surface modified single molecules free-diffusion evidenced by fluorescence correlation spectroscopy.Journal of fluorescence · 2008Article
- Total internal reflection fluorescence correlation spectroscopy: effects of lateral diffusion and surface-generated fluorescence.Biophysical journal · 2008Article
- Supercritical angle fluorescence correlation spectroscopy.Biophysical journal · 2008Article
- Kinetics of complexin binding to the SNARE complex: correcting single molecule FRET measurements for hidden events.Biophysical journal · 2007Article
- Trends in fluorescence imaging and related techniques to unravel biological information.HFSP journal · 2007Article
- Dynamics of synaptic SfiI-DNA complex: single-molecule fluorescence analysis.Biophysical journal · 2007Article
- Lateral mobility of membrane-binding proteins in living cells measured by total internal reflection fluorescence correlation spectroscopy.Biophysical journal · 2006Article
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3 authors.
Funding
Abstract
Total internal reflection excitation used in combination with fluorescence correlation spectroscopy (TIR-FCS) is a method for characterizing the dynamic behavior and absolute concentrations of fluorescent molecules near or at the interface of a planar substrate and a solution. In this work, we demonstrate for the first time the use of TIR-FCS for examining the interaction kinetics of fluorescent ligands in solution which specifically and reversibly associate with receptors in substrate-supported planar membranes. Fluorescence fluctuation autocorrelation functions were obtained for a fluorescently labeled IgG reversibly associating with the mouse receptor FcgammaRII, which was purified and reconstituted into substrate-supported planar membranes. Data were obtained as a function of the IgG solution concentration, the Fc receptor surface density, the observation area size, and the incident intensity. Best fits of the autocorrelation functions to appropriate theoretical forms gave measures of the average surface density of bound IgG, the local solution concentration of IgG, the kinetic rate constant for surface dissociation, and the rate of diffusion through the depth of the evanescent field. The average number of observed fluorescent molecules, both in solution and bound to the surface, scaled with the solution concentration of IgG, observation area size, and Fc receptor surface density as expected. The dissociation rate constant and rate of diffusion through the evanescent field agree with previous results, and all measured parameters were independent of the incident intensity.
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