Proceedings of September Meeting 2016 (NFM16)
Publication date: 14th June 2016
Imaging biological systems at the single-molecule level reveals protein motions and spatial heterogeneities that are lost in ensemble imaging experiments, but an ongoing challenge is the development of luminescent probes with the brightness, stability, continuous emission, and biocompatibility necessary for single-molecule microscopy. Semiconductor quantum dots (QDs) have proven to be superior single-particle probes compared to organic or genetically encoded fluorophores, but they are limited by difficulties in protein targeting, their larger sizes, and certain optical properties such as on-off blinking. Here we report compact aqueous quasi-type II CdSe/CdS QDs with significantly improved labeling and single-molecule optical properties. Single-molecule analysis of aqueous QDs shows a 99% on rate, ~3-fold higher single-QD quantum efficiency than standard CdSe/ZnS QDs, and 350 million photons detected over the life of the QD before photobleaching. To reduce QD size, we have synthesized novel covalent protein labeling ligands (i.e., SNAP tags) that are optimized for nanoparticle use and are over an order of magnitude more efficient than existing ligands. The improved protein labeling efficiency allows live-cell kinesin imaging and dual-color QD labeling of pairs of kinesin heads to quantitatively track motor protein movements.
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