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Related Experiment Video

Updated: Oct 16, 2025

A Protocol for Real-time 3D Single Particle Tracking
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Particles 3D tracking with large axial depth by using the 2π-DH-PSF.

Hangfeng Li, Famin Wang, Tongda Wei

    Optics Letters
    |October 15, 2021
    PubMed
    Summary

    We developed a novel 2π-double-helix point spread function (2π-DH-PSF) for advanced 3D nanoparticle imaging. This method significantly improves axial tracking range and localization accuracy in microscopy.

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    Area of Science:

    • Optics and Photonics
    • Biophysics
    • Nanotechnology

    Background:

    • Accurate three-dimensional (3D) localization of nanoparticles is crucial for various scientific fields.
    • Conventional double-helix point spread functions (DH-PSFs) offer 3D imaging capabilities but have limitations in axial range and rotation.
    • Developing advanced PSF engineering techniques is essential for enhancing microscopy performance.

    Purpose of the Study:

    • To introduce and validate a novel 2π-double-helix point spread function (2π-DH-PSF) for improved 3D super-resolution microscopy.
    • To demonstrate the enhanced axial tracking range and localization accuracy of the proposed 2π-DH-PSF.
    • To evaluate the performance of 2π-DH-PSF in measuring nanoparticle diffusion dynamics.

    Main Methods:

    • Utilized the Fresnel zone approach to design a 2π-DH-PSF capable of rotating 2π radians.

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  • Implemented the 2π-DH-PSF in a 100× microscopy system (NA=1.4, λ=514nm) with 16 Fresnel zones.
  • Measured the diffusion coefficient of nanospheres in varying glycerol concentrations using the 2π-DH-PSF.

    • Main Results:

    • Achieved particle tracking in an axial range of 10 µm.
    • Demonstrated superior 3D localization imaging performance for nanoparticles with the 2π-DH-PSF.
    • Obtained diffusion coefficient measurements with an error within 10% compared to theoretical values.
    • Showcased a potential rotation angle of 4π radians when combined with defocus phase, four times that of conventional DH-PSF.

    Conclusions:

    • The proposed 2π-DH-PSF significantly enhances axial tracking range and localization accuracy in 3D imaging.
    • The 2π-DH-PSF offers superior performance for nanoparticle tracking and diffusion analysis.
    • This advanced PSF engineering approach holds great promise for future developments in super-resolution microscopy and biophysical studies.