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    We developed a new adaptive optics (AO) method for Bessel-focus two-photon fluorescence microscopy (2PFM) with annular pupils. This modal focal AO (MFAO) simplifies optical configuration while maintaining high performance for clearer biological imaging.

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    Area of Science:

    • Biomedical Optics
    • Microscopy
    • Optical Engineering

    Background:

    • Adaptive optics (AO) enhances microscopy resolution by correcting optical aberrations.
    • AO is established for circular pupils but underexplored for non-circular pupils like those in Bessel-focus two-photon fluorescence microscopy (2PFM).

    Purpose of the Study:

    • To introduce and validate a modal focal AO (MFAO) method for Bessel-focus 2PFM with annular pupils.
    • To simplify AO implementation in microscopy systems with non-circular pupils.

    Main Methods:

    • Developed a modal focal AO (MFAO) method using Zernike annular polynomials.
    • Implemented aberration correction using a spatial light modulator at the objective's focal plane.
    • Validated MFAO with artificial and sample-induced aberrations, and in vivo imaging.

    Main Results:

    • MFAO achieved performance comparable to existing zonal AO methods.
    • The simplified optical configuration of MFAO offers practical advantages.
    • Successfully applied MFAO for in vivo imaging of zebrafish larvae and mouse brains.

    Conclusions:

    • MFAO expands modal AO applications to annular pupils.
    • This method simplifies aberration correction at the objective focal plane.
    • MFAO represents a significant advancement for AO implementation in microscopy.