Covalent Protein Labeling and Improved Single-Molecule Optical Properties of Aqueous CdSe/CdS Quantum Dots

Wichner, Sara M.; Mann, Victor R.; Powers, Alexander S.; Segal, Maya A.; Mir, Mustafa; Bandaria, Jigar N.; DeWitt, Mark A.; Darzacq, Xavier; Yildiz, Ahmet; Cohen, Bruce E.

doi:10.1021/acsnano.7b01470

Covalent Protein Labeling and Improved Single-Molecule Optical Properties of Aqueous CdSe/CdS Quantum Dots

Journal Article · Thu Jun 15 00:00:00 EDT 2017 · ACS Nano

DOI:https://doi.org/10.1021/acsnano.7b01470· OSTI ID:1973230

Wichner, Sara M. ^[1]; Mann, Victor R. ^[2]; Powers, Alexander S. ^[2]; Segal, Maya A. ^[1]; Mir, Mustafa ^[1]; Bandaria, Jigar N. ^[1]; DeWitt, Mark A. ^[1]; Darzacq, Xavier ^[1]; Yildiz, Ahmet ^[1]; ^[2]

University of California, Berkeley, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States). Molecular Foundry
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States). Molecular Foundry

Semiconductor quantum dots (QDs) have proven to be superior probes for single-molecule imaging compared to organic or genetically encoded fluorophores, but they are limited by difficulties in protein targeting, their larger size, and on-off blinking. We report compact aqueous CdSe/CdS QDs with significantly improved bioconjugation efficiency and superior single-molecule optical properties. We have synthesized covalent protein labeling ligands (i.e., SNAP tags) that are optimized for nanoparticle use, and QDs functionalized with these ligands label SNAP-tagged proteins ~10-fold more efficiently than existing SNAP ligands. Single-molecule analysis of these QDs shows 99% of time spent in the fluorescent on-state, ~4-fold higher quantum efficiency than standard CdSe/ZnS QDs, and 350 million photons detected before photobleaching. Bright signals of these QDs enable us to track the stepping movement of a kinesin motor in vitro, and the improved labeling efficiency enables tracking of single kinesins in live cells.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: California Institute for Regenerative Medicine; Miller Institute; National Institutes of Health (NIH); National Science Foundation (NSF); USDOE Office of Science (SC), Basic Energy Sciences (BES). Materials Sciences & Engineering Division (MSE)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1973230

Journal Information:: ACS Nano, Journal Name: ACS Nano Journal Issue: 7 Vol. 11; ISSN 1936-0851

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
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Covalent Protein Labeling and Improved Single-Molecule Optical Properties of Aqueous CdSe/CdS Quantum Dots

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