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A Protocol for Real-time 3D Single Particle Tracking

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

Nanoscale three-dimensional single particle tracking by light-sheet-based double-helix point spread function

Bin Yu, Jie Yu, Weihai Li

|February 3, 2016

Summary

This summary is machine-generated.

We improved nanoscale 3D tracking of single molecules using double-helix point spread function (DH-PSF) microscopy combined with light-sheet illumination. This method enhances precision by reducing background noise in thick biological samples.

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Last Updated: Mar 26, 2026

A Protocol for Real-time 3D Single Particle Tracking

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Published on: January 3, 2018

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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

Area of Science:

Biophysics
Optical Microscopy
Cell Biology

Background:

Double-helix point spread function (DH-PSF) microscopy enables nanoscale 3D single-molecule localization and tracking in living cells.
Localization precision is often limited in thick samples due to low signal-to-noise ratio from background fluorescence and low transfer function efficiency.

Purpose of the Study:

To enhance 3D single-particle tracking precision in DH-PSF microscopy.
To overcome limitations of background fluorescence in thick biological specimens.

Main Methods:

Combined DH-PSF microscopy with light-sheet illumination.
Utilized light-sheet illumination to minimize out-of-focus background fluorescence.
Acquired single fluorescent bead images and tracked particles in agarose solutions.

Main Results:

Achieved higher 3D localization accuracy compared to traditional epi-illumination.
Demonstrated successful tracking of single fluorescent beads in agarose, indicating potential for complex biological samples.
Significantly reduced background fluorescence through light-sheet integration.

Conclusions:

The combination of DH-PSF microscopy and light-sheet illumination offers a powerful approach for high-precision 3D single-particle tracking.
This technique holds promise for studying dynamic processes within complex biological environments.
Improved localization accuracy and background reduction pave the way for advanced live-cell imaging studies.