High-Precision Tracking with Non-blinking Quantum Dots Resolves Nanoscale Vertical Displacement

Date
2012-03-29
Authors
Marchuk, Kyle
Guo, Yijun
Sun, Wei
Vela, Javier
Vela, Javier
Fang, Ning
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Research Projects
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Ames Laboratory
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Chemistry
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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.

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<p>Reprinted (adapted) with permission from <em>Journal of the American Chemical Society</em> 134 (2012): 6108, doi:<a href="http://dx.doi.org/10.1021/ja301332t" target="_blank">10.1021/ja301332t</a>. Copyright 2012 American Chemical Society.</p>
Keywords
decay function, emission intensity, fluorescence imaging, fluorescence intensity decays, high-precision, incident angles, microtubules, nano scale, total internal reflection fluorescence, tracking application, vertical displacements, refractive index, semiconductor quantum dots, fluorescence, biotin, cadmium selenide, protein interaction, rotation, scanning angle total internal reflection fluorescence microscopy, biophysics
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