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    We developed a novel 3D interferometric fluorescent particle tracking method for high-resolution localization without sample scanning. This technique achieves nanometer resolution, proving effective in biological systems like tracking bacterial proteins.

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    Area of Science:

    • Biophysics
    • Soft Condensed Matter Physics
    • Microscopy

    Background:

    • Accurate particle tracking is crucial in biophysics and soft matter physics.
    • Existing methods often require sample scanning, limiting speed and applicability.

    Purpose of the Study:

    • To develop a novel 3D interferometric fluorescent particle tracking approach.
    • To achieve high-resolution particle localization without sample scanning.

    Main Methods:

    • Utilized a 3D interferometric microscopy technique.
    • Employed periodic shifting of interferometer phase to analyze interference patterns.
    • Tracked fluorescent beads and RecA protein in bacteria.

    Main Results:

    • Achieved nanometer-level localization resolution.
    • Demonstrated sub-5 nm axial localization error for a fluorescent bead at 5 Hz.
    • Validated compatibility with biological systems by tracking RecA protein in Bacillus subtilis.

    Conclusions:

    • The developed method offers accurate and rapid 3D particle tracking.
    • This technique eliminates the need for sample scanning, enhancing efficiency.
    • The approach is suitable for various applications in biophysics and soft matter research, including biological studies.