期刊
IMMUNE SYNAPSE: Methods and Protocols
卷 1584, 期 -, 页码 307-331出版社
HUMANA PRESS INC
DOI: 10.1007/978-1-4939-6881-7_18
关键词
Single-molecule; Bifunctional biomimetic surfaces; Nanofabrication; Self-assembly; Fluorescence microscopy
资金
- National Science Foundation (NSF) [CMMI-1300590]
- National Institutes of Health (NIH) Common Fund Nanomedicine program [PN2 EY016586]
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.
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