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Scanning Probe Microscopy in Nanoscience and Nanotechnology 3 pp 137–169Cite as

Single-Molecule Studies of Integrins by AFM-Based Force Spectroscopy on Living Cells

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Abstract

The characterization of cell adhesion between two living cells at the single-molecule level, i.e., between one adhesion receptor and its counter-receptor, appears to be an experimental challenge. Atomic force microscopy (AFM) can be used in its force spectroscopy mode to determine unbinding forces of a single pair of adhesion receptors, even with a living cell as a probe. This chapter provides an overview of AFM force measurements of the integrin family of cell adhesion receptors and their ligands. A focus is given to major integrins expressed on leukocytes, such as lymphocyte function-associated antigen 1 (LFA-1) and very late antigen 4 (VLA-4). These receptors are crucial for leukocyte trafficking in health and disease. LFA-1 and VLA-1 can be activated within the bloodstream from a low-affinity to a high-affinity receptor by chemokines in order to adhere strongly to the vessel wall before the receptor-bearing leukocytes extravasate. The experimental considerations needed to provide near-physiological conditions for a living cell and to be able to measure adequate forces at the single-molecule level are discussed in detail. AFM technology has been developed into a modern and extremely sensitive tool in biomedical research. It appears now that AFM force spectroscopy could enter, within a few years, medical applications in diagnosis and therapy of cancer and autoimmune diseases.

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Acknowledgements

The author gratefully acknowledges Hermann Gaub, Martin Benoit, Vincent Moy, Xiaohui (Frank) Zhang, Olga Vinogradova, and Wolfang Knoll for expert advice in the field of AFM and nanotechnology; Irving L. Weissman, Eugene C. Butcher, Marcus Hubbe, Bernhard Holzmann, Uwe Gosslar, James Campbell, Albert Zlotnik, Anja Müller, and Horst Kessler for support and material in the field of rolling, chemokines, and tumor stem cell biology;

Timothy A. Springer and Ronen Alon for the discussion; Heinz Höfler who fully supported the idea of the technology transfer of rolling experiments to AFM nanotechnology from its beginning, Matthias Maiwald and Markus Schneemann for the critical comments on the manuscript, and Stephan Bärtsch for the discussion of approaches in systems biology. Parts of this work were supported by stipends from the German Research Council (DFG, Bonn), an award of the German Cancer Research Center (DKFZ, Heidelberg), and major traveling grants from the Max Planck Society, the Technical University of Munich, the Center for NanoScience in Munich, as well as a Dean’s Fellowship from Stanford University and tuition sponsoring by Irv Weissman.

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  1. Institute of Pathology, Technical University of Munich, Munich, Germany

    Robert H. Eibl M.D.

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  1. , Nanoprobe Laboratory for Bio- &, Ohio State University, West 19th Avenue 201, Columbus, 43210-1142, Ohio, USA

    Bharat Bhushan

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Eibl, R.H. (2012). Single-Molecule Studies of Integrins by AFM-Based Force Spectroscopy on Living Cells. In: Bhushan, B. (eds) Scanning Probe Microscopy in Nanoscience and Nanotechnology 3. NanoScience and Technology. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-25414-7_6

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