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