Molecular Occupancy of Nanodot Arrays

Haogang Cai, Haguy Wolfenson, David Depoil, Michael L. Dustin, Michael Sheetz, Shalom J. Wind

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Single-molecule nanodot arrays, in which a biomolecule of choice (protein, nucleic acid, etc.) is bound to a metallic nanoparticle on a solid substrate, are becoming an increasingly important tool in the study of biomolecular and cellular interactions. We have developed an on-chip measurement protocol to monitor and control the molecular occupancy of nanodots. Arrays of widely spaced nanodots and nanodot clusters were fabricated on glass surfaces by nanolithography and functionalized with fluorescently labeled proteins. The molecular occupancy was determined by monitoring individual fluorophore bleaching events, while accounting for fluorescence quenching effects. We found that the occupancy can be interpreted as a packing problem, and depends on nanodot size and binding ligand concentration, where the latter is easily adjusted to compensate the flexibility of dimension control in nanofabrication. The results are scalable with nanodot cluster size, extending to large area close packed arrays. As an example, the nanoarray platform was used to probe the geometric requirement of T-cell activation at the single-molecule level.

Original languageEnglish (US)
Pages (from-to)4173-4183
Number of pages11
JournalACS Nano
Volume10
Issue number4
DOIs
StatePublished - Apr 26 2016
Externally publishedYes

Fingerprint

Proteins
Nanolithography
Molecules
T-cells
Fluorophores
Nucleic acids
Biomolecules
Bleaching
Nanotechnology
Nucleic Acids
proteins
Quenching
nanofabrication
Fluorescence
Chemical activation
Ligands
nucleic acids
bleaching
Nanoparticles
Glass

Keywords

  • electron beam lithography
  • fluorescence quenching
  • molecular occupancy
  • photobleaching
  • single-molecule assays
  • stoichiometry

ASJC Scopus subject areas

  • Materials Science(all)
  • Engineering(all)
  • Physics and Astronomy(all)

Cite this

Cai, H., Wolfenson, H., Depoil, D., Dustin, M. L., Sheetz, M., & Wind, S. J. (2016). Molecular Occupancy of Nanodot Arrays. ACS Nano, 10(4), 4173-4183. https://doi.org/10.1021/acsnano.5b07425

Molecular Occupancy of Nanodot Arrays. / Cai, Haogang; Wolfenson, Haguy; Depoil, David; Dustin, Michael L.; Sheetz, Michael; Wind, Shalom J.

In: ACS Nano, Vol. 10, No. 4, 26.04.2016, p. 4173-4183.

Research output: Contribution to journalArticle

Cai, H, Wolfenson, H, Depoil, D, Dustin, ML, Sheetz, M & Wind, SJ 2016, 'Molecular Occupancy of Nanodot Arrays', ACS Nano, vol. 10, no. 4, pp. 4173-4183. https://doi.org/10.1021/acsnano.5b07425
Cai H, Wolfenson H, Depoil D, Dustin ML, Sheetz M, Wind SJ. Molecular Occupancy of Nanodot Arrays. ACS Nano. 2016 Apr 26;10(4):4173-4183. https://doi.org/10.1021/acsnano.5b07425
Cai, Haogang ; Wolfenson, Haguy ; Depoil, David ; Dustin, Michael L. ; Sheetz, Michael ; Wind, Shalom J. / Molecular Occupancy of Nanodot Arrays. In: ACS Nano. 2016 ; Vol. 10, No. 4. pp. 4173-4183.
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