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Functionalization of AFM Tips and Supports for Molecular Recognition Force Spectroscopy and Recognition Imaging

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Atomic Force Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1886))

Abstract

Linking of sensor molecules (e.g., antibodies) to an AFM tip converts it into a biosensor by which single target molecules (e.g., antigens) can be detected and localized on the sample surface. Moreover, the mechanism of interaction can be studied by force spectroscopy if purified target molecules are linked to an ultra-flat surface, such as mica or silicon (nitride). Rapid imaging of the binding sites and force spectroscopy studies are greatly facilitated if 6–10 nm long polyethylene glycol (PEG) chains are used as flexible tethers between the sensor molecule and the tip. Here, we describe a set of methods by which a variety of proteins, oligonucleotides, or small molecules can be tethered to silicon (nitride) tips or to mica. Methods are included which afford site-specific and oriented coupling of the sensor molecules.

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Acknowledgements

This work was supported by the Austrian Science Funds (FWF project 15295), by the Austrian Research Promotion Agency (FFG project 819703 NSI-NABIOS), and by the EU-project Tips4Cells LSHG-CT-2005-512101.

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Authors and Affiliations

  1. Institute of Biophysics, Johannes Kepler University Linz, Linz, Austria

    A. Ebner, L. Wildling & H. J. Gruber

Authors
  1. A. Ebner
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  2. L. Wildling
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  3. H. J. Gruber
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Corresponding author

Correspondence to H. J. Gruber .

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Editors and Affiliations

  1. Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal

    Nuno C. Santos

  2. Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal

    Filomena A. Carvalho

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Ebner, A., Wildling, L., Gruber, H.J. (2019). Functionalization of AFM Tips and Supports for Molecular Recognition Force Spectroscopy and Recognition Imaging. In: Santos, N., Carvalho, F. (eds) Atomic Force Microscopy. Methods in Molecular Biology, vol 1886. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8894-5_7

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  • DOI: https://doi.org/10.1007/978-1-4939-8894-5_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8893-8

  • Online ISBN: 978-1-4939-8894-5

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