Small quantum dots conjugated to nanobodies as immunofluorescence probes for nanometric microscopy

Yong Wang, En Cai, Tobias Rosenkranz, Pinghua Ge, Kai Wen Teng, Sung Jun Lim, Andrew M. Smith, Hee Jung Chung, Frederick Sachs, William N. Green, Philip Gottlieb, Paul R. Selvin

Research output: Contribution to journalArticle

Abstract

Immunofluorescence, a powerful technique to detect specific targets using fluorescently labeled antibodies, has been widely used in both scientific research and clinical diagnostics. The probes should be made with small antibodies and high brightness. We conjugated GFP binding protein (GBP) nanobodies, small single-chain antibodies from llamas, with new ∼7 nm quantum dots. These provide simple and versatile immunofluorescence nanoprobes with nanometer accuracy and resolution. Using the new probes we tracked the walking of individual kinesin motors and measured their 8 nm step sizes; we tracked Piezo1 channels, which are eukaryotic mechanosensitive channels; we also tracked AMPA receptors on living neurons. Finally, we used a new super-resolution algorithm based on blinking of (small) quantum dots that allowed ∼2 nm precision.

Original languageEnglish (US)
Pages (from-to)2205-2211
Number of pages7
JournalBioconjugate Chemistry
Volume25
Issue number12
DOIs
StatePublished - Dec 17 2014

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

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  • Cite this

    Wang, Y., Cai, E., Rosenkranz, T., Ge, P., Teng, K. W., Lim, S. J., Smith, A. M., Chung, H. J., Sachs, F., Green, W. N., Gottlieb, P., & Selvin, P. R. (2014). Small quantum dots conjugated to nanobodies as immunofluorescence probes for nanometric microscopy. Bioconjugate Chemistry, 25(12), 2205-2211. https://doi.org/10.1021/bc5004179