Quantifying Reversible Surface Binding via Surface-Integrated Fluorescence Correlation Spectroscopy.

ABSTRACT: We present a simple and versatile single-molecule-based method for the accurate determination of binding rates to surfaces or surface bound receptors. To quantify the reversible surface attachment of fluorescently labeled molecules, we have modified previous schemes for fluorescence correlation spectroscopy with total internal reflection illumination (TIR-FCS) and camera-based detection. In contrast to most modern applications of TIR-FCS, we completely disregard spatial information in the lateral direction. Instead, we perform correlation analysis on a spatially integrated signal, effectively converting the illuminated surface area into the measurement volume. In addition to providing a high surface selectivity, our new approach resolves association and dissociation rates in equilibrium ov

SUBMITTER: Mucksch J 

PROVIDER: S-EPMC5946168 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature