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    This study introduces a phase-modulated spinning disk (PMSD) to improve structured illumination microscopy (SIM). The new PMSD-SIM technique enhances optical sectioning for faster, clearer live-cell imaging.

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    Area of Science:

    • Microscopy
    • Optical Physics
    • Biotechnology

    Background:

    • Structured illumination microscopy (SIM) offers high imaging speed for live-cell applications.
    • Conventional SIM lacks effective optical sectioning capabilities, limiting its use in complex biological samples.
    • Improving optical sectioning in SIM is crucial for enhanced image clarity and reduced out-of-focus blur.

    Purpose of the Study:

    • To develop a novel method for enhancing the optical sectioning capability of structured illumination microscopy.
    • To integrate a phase-modulated spinning disk (PMSD) into SIM for improved performance.
    • To enable real-time reconstruction of super-resolved images with higher contrast.

    Main Methods:

    • A phase-modulated spinning disk (PMSD) was designed, incorporating a pinhole array and a polymer layer for light phase modulation.
    • The PMSD was engineered to cancel the zeroth-order diffracted beam and generate a sharp lattice illumination pattern via interference.
    • The PMSD was utilized as a spatial filter in the detection path to reject out-of-focus light.

    Main Results:

    • The PMSD effectively enhanced the optical sectioning capability of SIM.
    • Approximately 80% of out-of-focus signals were rejected by the PMSD acting as a spatial filter.
    • Real-time optical reconstruction yielded super-resolved images with significantly improved contrast.

    Conclusions:

    • The novel PMSD approach substantially improves SIM's optical sectioning, addressing a key limitation.
    • This technique facilitates clearer, higher-contrast super-resolution live-cell imaging.
    • The PMSD design allows for straightforward upgrades to existing fluorescence microscopes, promoting wider adoption.