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Protein Dynamics in Living Cells

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Related Experiment Video

Updated: May 2, 2026

A Protocol for Real-time 3D Single Particle Tracking

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

Single-molecule tracking technology based on symmetric power exponential phases.

Famin Wang, Huijian Liu, Yunhai Zhang

|October 15, 2024

Summary

This summary is machine-generated.

We developed a novel symmetric power exponential phase for advanced 3D single-molecule imaging. This method enables precise nanoscale localization and tracking with adjustable probing depth.

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Last Updated: May 2, 2026

A Protocol for Real-time 3D Single Particle Tracking

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Published on: January 3, 2018

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

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Published on: September 5, 2019

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Published on: November 23, 2021

Area of Science:

Optics and Photonics
Biophysics
Nanotechnology

Background:

Accurate 3D single-molecule localization is crucial for understanding biological processes.
Existing methods face limitations in probing depth and precision.
Phase modulation offers a promising avenue for improving point-spread function engineering.

Purpose of the Study:

To design and validate a novel phase-modulated point-spread function for enhanced 3D nanoscale localization.
To enable precise 3D single-molecule tracking and localization imaging with adjustable axial probing depth.

Main Methods:

Design of a symmetric power exponential phase.
Development of a phase-modulated point-spread function with steerable dual mainlobes.
Application of truncation filtering and a phase inversion-based optimization algorithm.
Experimental validation using multi-molecule 3D tracking at 10 µm axial depth.

Main Results:

The proposed design yields a point-spread function with two symmetrical mainlobes steerable towards the optical axis.
Adjustable axial probing depth was achieved by manipulating specific design parameters.
Optimization strategies significantly reduced sidelobes and enhanced localization precision.
Successful multi-molecule 3D tracking demonstrated the method's efficacy at a 10 µm depth.

Conclusions:

The symmetric power exponential phase design provides a robust platform for nanoscale 3D localization.
This approach significantly advances capabilities for 3D single-molecule tracking and imaging.
The method offers tunable axial probing depth, enhancing its versatility in biological applications.