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Updated: May 7, 2026

A Protocol for Real-time 3D Single Particle Tracking

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

Approach to multiparticle parallel tracking in thick samples with three-dimensional nanoresolution.

Danni Chen, Bin Yu, Heng Li

|October 2, 2013

Summary

This summary is machine-generated.

A new microscopy method, distorted grating (DG) and double-helix point spread function (DH-PSF) combination microscopy (DDCM), enables simultaneous tracking of multiple particles in thick samples. This technique achieves 3D nanolocation with high precision across large depths, crucial for live cell imaging.

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Published on: October 1, 2014

Area of Science:

Optical microscopy
Biophysics
Nanotechnology

Background:

Accurate 3D tracking of multiple particles in thick biological samples remains a challenge.
Existing methods often struggle with depth-dependent localization precision and speed.

Purpose of the Study:

To introduce a novel microscopy technique, DDCM, for high-precision, multiparticle parallel localization and tracking.
To demonstrate DDCM's capability in transparent samples exceeding cellular thickness (10 μm).

Main Methods:

Development of a specialized phase mask combining distorted grating (DG) and double-helix point spread function (DH-PSF) functionalities.
Simulation of time-lapse tracking for single particles (z-axis) and parallel tracking for multiple particles.
Comparison of DDCM's theoretical localization precision with DH-PSF and multifocus microscopy.

Main Results:

DDCM enables multiparticle parallel localization and tracking in samples >10 μm thick.
Single-snapshot localization and tracking with 3D nanoresolution are achieved.
DDCM exhibits nearly constant 3D localization precision over a >10 μm depth of field.

Conclusions:

DDCM significantly enhances 3D tracking capabilities in thick biological samples.
The method offers superior and consistent localization precision across large depths compared to existing techniques.
DDCM is poised to become a valuable tool for studying dynamic biological processes in living cells.