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Related Experiment Video

Updated: May 23, 2026

A Protocol for Real-time 3D Single Particle Tracking
10:16

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

High-precision tracking with non-blinking quantum dots resolves nanoscale vertical displacement.

Kyle Marchuk1, Yijun Guo, Wei Sun

  • 1Ames Laboratory, U.S. Department of Energy, and Department of Chemistry, Iowa State University, Ames, Iowa 50011, USA.

Journal of the American Chemical Society
|March 31, 2012
PubMed
Summary
This summary is machine-generated.

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Novel non-blinking quantum dots (NBQDs) enable precise 3D tracking and rotation analysis of microtubules. This advanced fluorescence microscopy technique achieves sub-10-nm vertical localization accuracy.

Area of Science:

  • Biophysics
  • Nanotechnology
  • Microscopy

Background:

  • Accurate 3D tracking of biological molecules is crucial for understanding cellular processes.
  • Conventional quantum dots suffer from blinking, limiting their utility in long-term, high-precision imaging.
  • Total internal reflection fluorescence microscopy (TIRFM) offers enhanced surface sensitivity but requires precise vertical positioning.

Purpose of the Study:

  • To develop and validate a novel method for high-precision 3D super-localization and tracking of microtubules.
  • To assess the performance of novel non-blinking quantum dots (NBQDs) in advanced fluorescence microscopy.
  • To demonstrate the capability of resolving microtubule self-rotation using NBQD fluorescence intensity profiles.

Main Methods:

  • Utilized novel non-blinking quantum dots (NBQDs) attached to microtubules under gliding assay conditions.

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3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles
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3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles

Published on: October 1, 2014

Related Experiment Videos

Last Updated: May 23, 2026

A Protocol for Real-time 3D Single Particle Tracking
10:16

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles
11:28

3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles

Published on: October 1, 2014

  • Employed an automated scanning-angle total internal reflection fluorescence microscope (SA-TIRFM).
  • Analyzed fluorescence intensity decay curves obtained by varying incident angles and evanescent-field thicknesses to determine vertical positions.

    • Main Results:

    • Achieved sub-10-nm localization precision in the vertical (Z) dimension.
    • Successfully resolved the self-rotation of kinesin-propelled microtubules within a vertical distance of approximately 50 nm.
    • Demonstrated the stability and non-blinking characteristics of NBQDs during high-precision tracking.

    Conclusions:

    • Novel non-blinking quantum dots are highly effective for 3D super-localization and high-precision tracking applications.
    • The SA-TIRFM technique combined with NBQDs provides unprecedented accuracy in determining molecular positions and dynamics.
    • This methodology opens new avenues for studying nanoscale biological processes with enhanced resolution and reliability.