title: Spatial Control of Biological Ligands on Surfaces Applied to T Cell Activation.
abstract: In this chapter, we present techniques, based on molecular-scale nanofabrication and selective self-assembly, for the presentation of biomolecules of interest (ligands, receptors, etc.) on a surface with precise spatial control and arbitrary geometry at the single-molecule level. Metallic nanodot arrays are created on glass coverslips and are then used as anchors for the immobilization of biological ligands via thiol linking chemistry. The nanodot size is controlled by both lithography and metallization. The reagent concentration in self-assembly can be adjusted to ensure single-molecule occupancy for a given dot size. The surrounding glass is backfilled by a protein-repellent layer to prevent nonspecific adsorption. Moreover, bifunctional surfaces are created, whereby a second ligand is presented on the background, which is frequently a requirement for simulating complex cellular functions involving more than one key ligand. This platform serves as a novel and powerful tool for molecular and cellular biology, e.g., to study the fundamental mechanisms of receptor-mediated signaling.
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, USA.
authors.forename: Haogang
authors.initials: H
authors.lastname: Cai
authors.affiliation: Kennedy Institute of Rheumatology, NDORMS, The University of Oxford, Oxford, UK.
authors.forename: David
authors.initials: D
authors.lastname: Depoil
authors.affiliation: Department of Pathology, Skirball Institute, New York University School of Medicine, New York, USA.
authors.forename: James
authors.initials: J
authors.lastname: Muller
authors.affiliation: Department of Biological Sciences, Columbia University, New York, USA.|National University of Singapore, Singapore, Singapore.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Sheetz
authors.affiliation: Kennedy Institute of Rheumatology, NDORMS, The University of Oxford, Oxford, UK.|Department of Pathology, Skirball Institute, New York University School of Medicine, New York, USA.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Dustin
authors.affiliation: Department of Applied Physics and Applied Mathematics, Columbia University, 500 W 120th St, New York, NY, 10027, USA. sw2128@columbia.edu.
authors.forename: Shalom J
authors.initials: SJ
authors.lastname: Wind
mesh_terms: D000818:Animals; D006801:Humans; D008213:Lymphocyte Activation; D046228:Microarray Analysis / Q000295:instrumentation* / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry*; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000737:chemistry* / Q000276:immunology
keywords: Bifunctional biomimetic surfaces; Fluorescence microscopy; Nanofabrication; Self-assembly; Single-molecule