title: High-precision tracking with non-blinking quantum dots resolves nanoscale vertical displacement.
abstract: Novel non-blinking quantum dots (NBQDs) were utilized in three-dimensional super-localization, high-precision tracking applications under an automated scanning-angle total internal reflection fluorescence microscope (SA-TIRFM). NBQDs were randomly attached to stationary microtubules along the radial axis under gliding assay conditions. By automatically scanning through a wide range of incident angles with different evanescent-field layer thicknesses, the fluorescence intensity decay curves were obtained. Fit with theoretical decay functions, the absolute vertical positions were determined with sub-10-nm localization precision. The emission intensity profile of the NBQDs attached to kinesin-propelled microtubules was used to resolve the self-rotation of gliding microtubules within a small vertical distance of ~50 nm. We demonstrate the applicability of NBQDs in high-precision fluorescence imaging experiments.
authors.affiliation: Ames Laboratory, U.S. Department of Energy, and Department of Chemistry, Iowa State University, Ames, Iowa 50011, USA.
authors.forename: Kyle
authors.initials: K
authors.lastname: Marchuk
authors.forename: Yijun
authors.initials: Y
authors.lastname: Guo
authors.forename: Wei
authors.initials: W
authors.lastname: Sun
authors.forename: Javier
authors.initials: J
authors.lastname: Vela
authors.forename: Ning
authors.initials: N
authors.lastname: Fang
mesh_terms: D000818:Animals; D001331:Automation; D001703:Biophysics / Q000379:methods; D006801:Humans; D021621:Imaging, Three-Dimensional; D016547:Kinesins / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods; D008870:Microtubules / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry; D036103:Nanotechnology / Q000379:methods; D045663:Quantum Dots; D015203:Reproducibility of Results; D012666:Semiconductors
chemical_list: D016547:Kinesins