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Experimental validation of a customized phase mask designed to enable efficient computational optical sectioning

Nurmohammed Patwary, Hasti Shabani, Ana Doblas

    Applied Optics
    |April 5, 2017
    PubMed
    Summary
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    Wavefront-encoded computational optical sectioning microscopy (WFE-COSM) uses a novel phase mask to reduce depth-induced aberrations. This imaging technique achieves over twofold performance improvement for deep tissue imaging compared to conventional methods.

    Area of Science:

    • Microscopy and Imaging Technologies
    • Computational Optics
    • Biomedical Engineering

    Background:

    • Depth-induced aberrations limit optical sectioning microscopy performance in thick samples.
    • Conventional methods often require computationally intensive depth-variant algorithms for image restoration.
    • Developing robust imaging systems less sensitive to axial position is crucial for deep tissue visualization.

    Purpose of the Study:

    • To investigate wavefront-encoded computational optical sectioning microscopy (WFE-COSM) using a square cubic (SQUBIC) phase mask.
    • To assess the system's reduced sensitivity to depth-induced aberrations.
    • To enable computationally efficient image restoration for large volume imaging.

    Main Methods:

    • Fabrication and characterization of a SQUBIC phase mask with 75% fidelity.

    Related Experiment Videos

  • Integration of the phase mask into a commercial wide-field microscope to create the WFE-COSM system.
  • Comparative analysis of theoretical and experimental point spread functions (PSFs) and imaging of spherical shells at various depths.

    • Main Results:

    • The WFE-COSM system exhibits a point spread function (PSF) with slower variation with depth compared to conventional systems.
    • PSF and imaging models showed a 99% correlation with experimental data for spherical shell imaging.
    • Experimental space-invariant (SI) restoration demonstrated over twofold performance improvement up to 65 μm depth.

    Conclusions:

    • WFE-COSM with a SQUBIC phase mask effectively reduces depth-induced aberrations.
    • The system allows for computationally efficient space-invariant (SI) image restoration.
    • This approach significantly enhances imaging performance in deep biological samples.