Quantitative analyses of cell adhesion strength.

Abstract

Biochemical and mechanical analyses of integrin-mediated cell adhesion have been limited by the inability to apply controlled detachment forces and the inherent complexities of the adhesive process, including cell spreading, integrin clustering, cytoskeletal interactions, and non-uniformly distributed focal complexes. A comprehensive set of techniques to analyze mechanical and biochemical events at the cell-extracellular matrix interface is presented. The spinning disk assay provides a rigorous hydrodynamic assay to measure adhesion strength. Crosslinking/extraction and wet-cleaving biochemical approaches isolate and quantify integrins bound to their ligands and adhesive components recruited to focal adhesions. These techniques provide an experimental framework for the rigorous analysis of cell adhesion.