Applications of Atomic Force Microscopy for Adhesion Force Measurements in Mechanotransduction.
Applications of Atomic Force Microscopy for Adhesion Force Measurements in Mechanotransduction.
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DOI：
10.1007/978-1-4939-8591-3_30
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发表时间：
2018
影响因子：
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通讯作者：
A. Trache;Leike Xie;Huang-Chieh Huang-Huang-Chieh-Huang-93294268;V. Glinsky;G. Meininger
中科院分区：
文献类型：
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作者：
A. Trache;Leike Xie;Huang-Chieh Huang-Huang-Chieh-Huang-93294268;V. Glinsky;G. Meininger
活细胞之间的黏附相互作用或配体-受体相互作用可以用原子力显微镜(AFM)在分子水平上进行研究。粘附力的测量是用功能化的原子力显微镜探针进行的。为了测量单个配体-受体的相互作用，具有锥体尖端的悬臂被生物识别的配体(例如细胞外基质蛋白)功能化。然后将配体功能化的探针与培养中的细胞接触，以研究各自的探针结合的配体与内源性表达的细胞表面受体(例如整合素或其他黏附受体)之间的黏附。在设计用来检测细胞间粘连的实验中，单个细胞被连接到一个无端悬臂上，然后悬臂与其他培养的细胞接触。测力曲线被记录以确定断裂探针结合的配体和受体之间的离散粘连所需的力，或确定细胞-细胞接触处的总粘附力。在这里，我们描述了测量(A)纤维连接蛋白和α5β1整合素，以及(B)乳腺癌细胞和骨髓内皮细胞之间粘连的程序。
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.