title: Increasing throughput of AFM-based single cell adhesion measurements through multisubstrate surfaces.
abstract: Mammalian cells regulate adhesion by expressing and regulating a diverse array of cell adhesion molecules on their cell surfaces. Since different cell types express distinct sets of cell adhesion molecules, substrate-specific adhesion is cell type- and condition-dependent. Single-cell force spectroscopy is used to quantify the contribution of cell adhesion molecules to adhesion of cells to specific substrates at both the cell and single molecule level. However, the low throughput of single-cell adhesion experiments greatly limits the number of substrates that can be examined. In order to overcome this limitation, segmented polydimethylsiloxane (PDMS) masks were developed, allowing the measurement of cell adhesion to multiple substrates. To verify the utility of the masks, the adhesion of four different cell lines, HeLa (Kyoto), prostate cancer (PC), mouse kidney fibroblast and MDCK, to three extracellular matrix proteins, fibronectin, collagen I and laminin 332, was examined. The adhesion of each cell line to different matrix proteins was found to be distinct; no two cell lines adhered equally to each of the proteins. The PDMS masks improved the throughput limitation of single-cell force spectroscopy and allowed for experiments that previously were not feasible. Since the masks are economical and versatile, they can aid in the improvement of various assays.
authors: Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, 4058 Basel, Switzerland ; Center for Precision Engineering, Harbin Institute of Technology, Harbin 150001, China.
authors: Miao
authors: M
authors: Yu
authors: Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, 4058 Basel, Switzerland.
authors: Nico
authors: N
authors: Strohmeyer
authors: Center for Precision Engineering, Harbin Institute of Technology, Harbin 150001, China.
authors: Jinghe
authors: J
authors: Wang
authors: Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, 4058 Basel, Switzerland.
authors: Daniel J
authors: DJ
authors: Müller
authors: Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, 4058 Basel, Switzerland.
authors: Jonne
authors: J
authors: Helenius
keywords: HeLa; MDCK; PC3; atomic force microscopy; cell adhesion; collagen I; fibroblasts; fibronectin; laminin; single cell assay; single cell force spectroscopy