Applications of Atomic Force Microscopy for Adhesion Force Measurements in Mechanotransduction
Methods in molecular biology, January 2018
Andreea Trache, Leike Xie, Huang, Vladislav V. Glinsky, Gerald A. Meininger, Huang Huang, Trache, Andreea, Xie, Leike, Huang, Huang, Glinsky, Vladislav V., Meininger, Gerald A.
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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