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Quantifying cellular adhesion to extracellular matrix components by single-cell force spectroscopy

Muller, Daniel J ; Friedrichs, Jens ; Helenius, Jonne

Nature protocols, 2010-07, Vol.5 (7), p.1353-1361 [Periódico revisado por pares]

England: Nature Publishing Group

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Título:
Quantifying cellular adhesion to extracellular matrix components by single-cell force spectroscopy
Autor: Muller, Daniel J ; Friedrichs, Jens ; Helenius, Jonne
Assuntos: Adhesives ; Atomic force microscopy ; Cell adhesion ; Cell adhesion & migration ; Cell Adhesion - physiology ; Cell research ; Collagen ; Collagen Type I ; Extracellular matrix ; Extracellular Matrix - metabolism ; Extracellular Matrix - physiology ; Extracellular Matrix - ultrastructure ; HeLa Cells ; Humans ; Integrins - metabolism ; Integrins - ultrastructure ; Ligands ; Methods ; Microscopy ; Microscopy, Atomic Force - instrumentation ; Microscopy, Atomic Force - methods ; Observations ; Physiology ; Protein Binding - physiology ; Proteins ; Spectrum analysis
É parte de: Nature protocols, 2010-07, Vol.5 (7), p.1353-1361
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SourceType-Scholarly Journals-1
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Descrição: Atomic force microscopy (AFM)-based single-cell force spectroscopy (SCFS) enables the quantitative study of cell adhesion under physiological conditions. SCFS probes adhesive interactions of single living cells with substrates such as extracellular matrix (ECM) proteins and other cells. Here we present a protocol to study integrin-mediated adhesion of HeLa cells to collagen type I using SCFS. We describe procedures for (i) functionalization of AFM cantilevers with the lectin concanavalin A and supports with collagen, (ii) cell handling and attachment to the AFM cantilever, (iii) measurement of adhesion forces and (iv) data analysis and interpretation. Although designed to measure HeLa cell adhesion to collagen, the protocol can be modified for other cell lines and ECM proteins. Compared with other SCFS assays (for example, optical tweezer, biomembrane force probe), AFM-based SCFS has a more versatile force detection range, and it can therefore be used to address a broader range of biological questions. The protocol can be completed in 2-3 d.
Editor: England: Nature Publishing Group
Idioma: Inglês

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Quantifying cellular adhesion to extracellular matrix components by single-cell force spectroscopy

Muller, Daniel J ; Friedrichs, Jens ; Helenius, Jonne

England: Nature Publishing Group

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