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  Quantifying Reversible Surface Binding via Surface-Integrated Fluorescence Correlation Spectroscopy

Mücksch, J., Blumhardt, P., Strauss, M. T., Petrov, E. P., Jungmann, R., & Schwille, P. (2018). Quantifying Reversible Surface Binding via Surface-Integrated Fluorescence Correlation Spectroscopy. Nano Letters, 18(5), 3185-3192. doi:10.1021/acs.nanolett.8b00875.

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 Creators:
Mücksch, Jonas1, Author              
Blumhardt, Philipp1, Author              
Strauss, Maximilian T.2, Author              
Petrov, Eugene P.1, Author              
Jungmann, Ralf2, Author              
Schwille, Petra1, Author              
Affiliations:
1Schwille, Petra / Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565169              
2Jungmann, Ralf / Molecular Imaging and Bionanotechnology, Max Planck Institute of Biochemistry, Max Planck Society, ou_2149679              

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Free keywords: TOTAL INTERNAL-REFLECTION; SINGLE-MOLECULE KINETICS; SUPERRESOLUTION MICROSCOPY; SOLID/LIQUID INTERFACES; CROSS-CORRELATION; DNA-PAINT; DIFFUSION; PROTEINS; BILAYER; CELLChemistry; Science & Technology - Other Topics; Materials Science; Physics; Total internal reflection fluorescence correlation spectroscopy (TIR-FCS); surface binding kinetics; binding rates; DNA hybridization; DNA-PAINT;
 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 over a wide range of time scales. We chose the transient hybridization of fluorescently labeled single-stranded DNA to the complementary handles of surface-immobilized DNA origami structures as a reliable and well-characterized test system. We varied the number of base pairs in the duplex, yielding different binding times in the range of hundreds of milliseconds to tens of seconds, allowing us to quantify the respective surface affinities and binding rates.

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Language(s): eng - English
 Dates: 2018
 Publication Status: Published in print
 Pages: 8
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Project name : This work was supported in part by the DFG through the Emmy Noether Program (DFG JU 2957/1-1), the SFB 1032 (Nanoagents for spatiotemporal control of molecular and cellular reactions, Project A11), the ERC through an ERC Starting Grant (MolMap, Grant agreement no. 680241), the Max Planck Society, the Max Planck Foundation, and the Center for Nanoscience (CeNS) to R.J.
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Title: Nano Letters
  Abbreviation : Nano Lett.
Source Genre: Journal
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Publ. Info: Washington, DC : American Chemical Society
Pages: - Volume / Issue: 18 (5) Sequence Number: - Start / End Page: 3185 - 3192 Identifier: ISSN: 1530-6984
CoNE: https://pure.mpg.de/cone/journals/resource/110978984570403