Spatial control of biological ligands on surfaces applied to T cell activation

Haogang Cai, David Depoil, James Muller, Michael Sheetz, Michael L. Dustin, Shalom J. Wind

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

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.

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Keywords

  • Bifunctional biomimetic surfaces
  • Fluorescence microscopy
  • Nanofabrication
  • Self-assembly
  • Single-molecule

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Cai, H., Depoil, D., Muller, J., Sheetz, M., Dustin, M. L., & Wind, S. J. (2017). Spatial control of biological ligands on surfaces applied to T cell activation. In Methods in Molecular Biology (pp. 307-331). (Methods in Molecular Biology; Vol. 1584). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6881-7_18