Abstract
Adhesive interactions between living cells or ligand-receptor interactions can be studied at the molecular level using atomic force microscopy (AFM). Adhesion force measurements are performed with functionalized AFM probes. In order to measure single ligand-receptor interactions, a cantilever with a pyramidal tip is functionalized with a bio-recognized ligand (e.g., extracellular matrix protein). The ligand-functionalized probe is then brought into contact with a cell in culture to investigate adhesion between the respective probe-bound ligand and endogenously expressed cell surface receptors (e.g., integrins or other adhesion receptor). For experiments designed to examine cell-cell adhesions, a single cell is attached to a tipless cantilever which is then brought into contact with other cultured cells. Force curves are recorded to determine the forces necessary to rupture discrete adhesions between the probe-bound ligand and receptor, or to determine total adhesion force at cell-cell contacts. Here, we describe the procedures for measuring adhesions between (a) fibronectin and α5β1 integrin, and (b) breast cancer cells and bone marrow endothelial cells.
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Acknowledgments
This work was supported by NIH P01HL095486 (to G.A. Meininger), NIH R01CA160461 (to V.V. Glinsky), and VA BLR&D Service Award 1I01BX000609 (to V.V. Glinsky).
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Trache, A., Xie, L., Huang, H., Glinsky, V.V., Meininger, G.A. (2018). Applications of Atomic Force Microscopy for Adhesion Force Measurements in Mechanotransduction. In: Lyubchenko, Y. (eds) Nanoscale Imaging. Methods in Molecular Biology, vol 1814. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8591-3_30
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DOI: https://doi.org/10.1007/978-1-4939-8591-3_30
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