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Updated: May 12, 2026

A Protocol for Real-time 3D Single Particle Tracking

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

Differential interferometric particle tracking on the subnanometer- and submillisecond-scale.

Dennis Müller¹, Dieter R Klopfenstein, Rainer G Ulbrich

¹IV. Physikalisches Institut, Georg-August-Universität Göttingen, Friedrich-Hund-Platz 1, 37077 Göttingen, Germany. dmueller@ph4.physik.uni-goettingen.de

|April 3, 2013

Summary

We developed a novel interferometric method to precisely track subwavelength particle movement. This technique successfully resolved kinesin

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Published on: October 1, 2014

Area of Science:

Biophysics
Nanotechnology
Optical Physics

Background:

Microscopic particle movement is crucial for understanding biological processes.
Existing methods often lack the required spatial and temporal resolution.
Microscope stage drift can introduce artifacts in high-resolution measurements.

Purpose of the Study:

To develop a high-resolution interferometric method for measuring subwavelength particle motion.
To eliminate microscope stage drift using a differential measurement technique.
To apply the method to observe molecular motor activity.

Main Methods:

Utilized an upright microscope with laser dark field illumination (532nm, 30mW).
Integrated a modified Mach-Zehnder interferometer at the microscope's camera exit.
Combined scattered light from probe and reference particles to generate interferograms.
Probed interferograms with avalanche photodiodes for subnanometer motion detection.

Main Results:

Achieved high spatial resolution of 0.9nm and temporal resolution of 200μs.
Successfully eliminated microscope stage drift effects.
Resolved the 8-nm steps of kinesin motor proteins moving along microtubules at high ATP concentrations without external forces.

Conclusions:

The developed interferometric method offers unprecedented resolution for tracking nanoscale movements.
This technique is suitable for studying motor proteins like kinesin in biological systems.
Enables direct observation of molecular motor stepping under physiological conditions.