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    Fourier phase microscopy (FPM) accurately measures microscopic bead sizes and classifies cells by analyzing scatterer size distribution. This quantitative phase imaging method offers high precision for biological and material science applications.

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    Area of Science:

    • Optical microscopy
    • Biophysics
    • Materials science

    Background:

    • Quantitative phase imaging (QPI) methods are crucial for label-free cell analysis.
    • Fourier phase microscopy (FPM) combines features of on-axis and off-axis QPI for enhanced angular scattering analysis.

    Purpose of the Study:

    • To demonstrate the first application of FPM for precise measurement of single-micrometer bead diameters.
    • To classify cells based on alterations in scatterer size distribution using FPM.
    • To validate FPM's accuracy against Mie theory and manufacturer specifications.

    Main Methods:

    • Utilized Fourier phase microscopy (FPM), a quantitative phase imaging technique.
    • Employed planar illumination and common-path geometry for optimal angular scattering.
    • Analyzed angular scattering patterns from 1 µm polystyrene beads and macrophages.

    Main Results:

    • FPM achieved high-fidelity angular scattering patterns consistent with Mie theory for polystyrene beads.
    • Accurate bead diameter estimations with precision in the tens of nanometers were obtained.
    • Detected scattering changes in macrophages during antibody-dependent cellular phagocytosis, indicating internal material presence.

    Conclusions:

    • FPM is a robust and precise tool for sub-micrometer particle sizing.
    • FPM enables label-free cell classification by detecting intracellular changes via scattering.
    • The FPM system is stable, easy to align, and suitable for various angular-domain applications.