title: Fabricating surfaces with distinct geometries and different combinations of cell adhesion proteins.
abstract: A step-by-step procedure is described for functionalizing the surface of a glass coverslip so that a single cell contacts distinct patterns of extracellular matrix and cell-cell adhesion proteins. This dual-micropatterned substratum is accomplished through a two-step process. First, extracellular matrix (ECM) is microcontact-printed onto a silanized glass surface using electron beam lithography, etched resist-coated wafers, and Polydimethylsiloxane (PDMS) stamps of differing geometries. Then, non-ECM-coated surfaces are incubated sequentially with biotin, NeutrAvidin, and biotinylated Protein A to attach Fc-cadherin fusion proteins, Fc, or PEG. Cells are seeded at low density onto the functionalized surface for single-cell analysis of protein recruitment/turnover and cellular motility.
authors.affiliation: Cancer Biology Program, Stanford University, Stanford, CA, USA.
authors.forename: Molly
authors.initials: M
authors.lastname: Lowndes
authors.forename: W James
authors.initials: WJ
authors.lastname: Nelson
mesh_terms: D001360:Avidin / Q000737:chemistry; D002448:Cell Adhesion / Q000235:genetics*; D015815:Cell Adhesion Molecules / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D059010:Single-Cell Analysis / Q000379:methods*; D013205:Staphylococcal Protein A / Q000737:chemistry; D013499:Surface Properties
chemical_list: D015815:Cell Adhesion Molecules; D011506:Proteins; D013205:Staphylococcal Protein A; C093483:neutravidin; D001360:Avidin