Chapter title |
Applications of Atomic Force Microscopy for Adhesion Force Measurements in Mechanotransduction
|
---|---|
Chapter number | 30 |
Book title |
Nanoscale Imaging
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8591-3_30 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8590-6, 978-1-4939-8591-3
|
Authors |
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. |
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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