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Fluorescence phase-shifting interferometry for axial single particle tracking: a numerical simulation study.

E Arbel, A Praiz, A Bilenca

    Optics Express
    |October 17, 2014
    PubMed
    Summary
    This summary is machine-generated.

    We introduce fluorescence phase-shifting interferometry (FPSI) for precise axial single particle tracking. Spatial FPSI offers improved precision for diffusion, while temporal FPSI excels in localizing slow linear movements.

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    Area of Science:

    • Biophysics
    • Optical Physics
    • Nanotechnology

    Background:

    • Axial tracking of single fluorescent probes is crucial for biological and physical sciences.
    • Existing methods face limitations in precision and depth-of-field for single particle tracking (SPT).

    Purpose of the Study:

    • To propose and analyze fluorescence phase-shifting interferometry (FPSI) for high-precision axial SPT.
    • To compare spatial and temporal FPSI methods for tracking particle trajectories along the z-axis.

    Main Methods:

    • Utilized fluorescence phase-shifting interferometry (FPSI), a photon-efficient self-interference technique.
    • Employed vectorial imaging analysis and Monte-Carlo simulations for diffusive and directed particle motions.
    • Collected four phase-shifted interferograms using a single camera for both spatial and temporal FPSI analysis.

    Main Results:

    • Spatial FPSI demonstrated up to a twofold improvement in trajectory reconstruction precision for diffusion (≤0.105 μm²/s) compared to temporal FPSI.
    • Spatial FPSI exhibited a fourfold reduced field-of-view relative to temporal FPSI.
    • Temporal FPSI was predicted to be superior for sufficiently slow random linear motions, yielding a smaller reconstruction error.

    Conclusions:

    • FPSI is a viable technique for high-precision axial single particle tracking over an extended depth-of-field.
    • The choice between spatial and temporal FPSI depends on the specific motion dynamics and desired trade-offs between precision and field-of-view.