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    This study introduces a vectorial model for Image Scanning Microscopy (ISM), improving resolution beyond the diffraction limit. The vectorial approach accurately accounts for polarization and aberrations, crucial for advanced high-resolution imaging.

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    Area of Science:

    • Optics and Photonics
    • Microscopy Techniques
    • Super-resolution Imaging

    Background:

    • Image Scanning Microscopy (ISM) offers super-resolution imaging beyond the diffraction limit.
    • Existing ISM models often rely on scalar diffraction theory, neglecting polarization, aberrations, and Stokes shift.
    • These neglected factors can significantly impact imaging accuracy and resolution.

    Purpose of the Study:

    • To develop a comprehensive vectorial Image Scanning Microscopy (ISM) point spread function (PSF) model.
    • To account for polarization, aberrations, and Stokes shift in ISM.
    • To analyze the discrepancies between scalar and vectorial ISM theories and their impact on resolution.

    Main Methods:

    • Development of a vectorial PSF model for ISM incorporating polarization, aberrations, and Stokes shift.
    • Comparative analysis of scalar and vectorial ISM theories.
    • Investigation of the influence of pinhole size and aberration strength on resolution.
    • Exploration of multiphoton excitation effects (2-photon and 3-photon) in ISM.

    Main Results:

    • The vectorial ISM model provides a more accurate representation of imaging phenomena.
    • Discrepancies between scalar and vectorial PSFs can reach 45 nm (30%) at NA 1.2.
    • Multiphoton excitation (2-photon and 3-photon) increases PSF FWHM by 20% and 28%, respectively.
    • Optimal sweep factors for 2-photon (1.22) and 3-photon (1.12) ISM differ from scalar theory predictions (2).

    Conclusions:

    • Vectorial modeling is essential for accurate Image Scanning Microscopy (ISM) analysis.
    • Polarization, aberrations, and Stokes shift critically affect ISM resolution.
    • The developed model enhances understanding and advancement of ISM for high-resolution applications.