title: Investigating cell-substrate and cell-cell interactions by means of single-cell-probe force spectroscopy.
abstract: Cell adhesion forces are typically a mixture of specific and nonspecific cell-substrate and cell-cell interactions. In order to resolve these phenomena, Atomic Force Microscopy appears as a powerful device which can measure cell parameters by means of manipulation of single cells. This method, commonly known as cell-probe force spectroscopy, allows us to control the force applied, the area of interest, the approach/retracting speed, the force rate, and the time of interaction. Here, we developed a novel approach for in situ cantilever cell capturing and measurement of specific cell interactions. In particular, we present a new setup consisting of two different half-surfaces coated either with recrystallized SbpA bacterial cell surface layer proteins (S-layers) or integrin binding Fibronectin, on which MCF-7 breast cancer cells are incubated. The presence of a clear physical boundary between both surfaces benefits for a quick detection of the region under analysis. Thus, quantitative results about SbpA-cell and Fibronectin-cell adhesion forces as a function of the contact time are described. Additionally, the importance of the cell spreading in cell-cell interactions has been studied for surfaces coated with two different Fibronectin concentrations: 20 μg/mL (FN20) and 100 μg/mL (FN100), which impact the number of substrate receptors. Microsc. Res. Tech. 80:124-130, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Nanobiotechnology, Institute for Biophysics, University of Natural Resources and Life Sciences Vienna (BOKU), Muthgasse 11, Vienna, 1190, Austria.
authors.forename: Alberto
authors.initials: A
authors.lastname: Moreno-Cencerrado
authors.affiliation: Department of Nanobiotechnology, Institute for Biophysics, University of Natural Resources and Life Sciences Vienna (BOKU), Muthgasse 11, Vienna, 1190, Austria.
authors.forename: Jagoba
authors.initials: J
authors.lastname: Iturri
authors.affiliation: Department of Engineering and Architecture, Università Degli Studi Di Trieste, via Valerio 6 - 34127, Trieste, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Pecorari
authors.affiliation: Cell Biology and Stem Cells Unit, CIC bioGUNE, Bizkaia Science and Technology Park, Derio, Spain.
authors.forename: Maria
authors.initials: M
authors.lastname: D M Vivanco
authors.affiliation: Department of Engineering and Architecture, Università Degli Studi Di Trieste, via Valerio 6 - 34127, Trieste, Italy.
authors.forename: Orfeo
authors.initials: O
authors.lastname: Sbaizero
authors.affiliation: Department of Nanobiotechnology, Institute for Biophysics, University of Natural Resources and Life Sciences Vienna (BOKU), Muthgasse 11, Vienna, 1190, Austria.
authors.forename: José L
authors.initials: JL
authors.lastname: Toca-Herrera
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D002448:Cell Adhesion; D002450:Cell Communication; D005353:Fibronectins / Q000737:chemistry; D006801:Humans; D061986:MCF-7 Cells; D018625:Microscopy, Atomic Force; D009004:Monosaccharide Transport Proteins / Q000737:chemistry; D011485:Protein Binding; D013499:Surface Properties
chemical_list: D001426:Bacterial Proteins; D005353:Fibronectins; D009004:Monosaccharide Transport Proteins; C120455:SbpA protein, bacteria
keywords: S-layer proteins; atomic force microscopy; cell adhesion; force spectroscopy; single-cell probe