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Related Experiment Video

Updated: Jun 24, 2026

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

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

Published on: October 1, 2014

3D particle trajectories observed by orthogonal tracking microscopy.

Matthew D McMahon¹, Andrew J Berglund, Peter Carmichael

¹Center for Nanoscale Science and Technology, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, USA. matthew.mcmahon@nist.gov

|March 25, 2009

Summary

High-speed 3D nanoparticle tracking is achieved using angled micromirrors. This method enables precise trajectory determination for potential real-time nanoparticle assembly applications.

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A Protocol for Real-time 3D Single Particle Tracking

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

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

Published on: October 1, 2014

A Protocol for Real-time 3D Single Particle Tracking

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

Area of Science:

Optics and Photonics
Nanotechnology
Biophysics

Background:

Accurate three-dimensional (3D) tracking of nanoparticles is crucial for understanding nanoscale phenomena.
Traditional methods often face limitations in speed, resolution, or computational complexity.
Developing faster and more precise nanoparticle tracking techniques is an ongoing challenge.

Purpose of the Study:

To demonstrate a novel method for high-resolution, high-speed 3D nanoparticle tracking.
To utilize angled micromirrors to capture multiple views of nanoparticles within an optical microscope.
To enable precise trajectory determination without complex computational algorithms.

Main Methods:

Introduction of angled micromirrors into the optical microscope's field of view.
Projection of reflected side-on views alongside the direct image of diffusing nanoparticles.
Fast, centroid-based image processing for determining 3D particle trajectories.
Tracking of 190 nm polystyrene particles with a 3 ms frame duration.

Main Results:

Achieved high-speed imaging at over 330 frames per second.
Demonstrated position measurement precision of less than 20 nm in 3D.
Image processing completed in approximately 1 ms per frame.
Successfully tracked nanoparticle trajectories in three dimensions.

Conclusions:

Angled micromirrors provide an effective means for high-speed, high-resolution 3D nanoparticle tracking.
The technique's speed and precision are suitable for advanced applications.
Potential for real-time feedback-controlled nanoparticle assembly with nanometer precision is established.