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Quantitative Optical Microscopy: Measurement of Cellular Biophysical Features with a Standard Optical Microscope
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Quantitative phase imaging and complex field reconstruction by pupil modulation differential phase contrast.

Hangwen Lu, Jaebum Chung, Xiaoze Ou

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    |November 10, 2016
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    Summary

    We developed pupil modulation differential phase contrast (PMDPC) for high-resolution quantitative phase imaging. This method achieves 1.73μm resolution and extends depth of field digitally.

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    Area of Science:

    • Optics and Photonics
    • Biomedical Imaging
    • Microscopy

    Background:

    • Differential phase contrast (DPC) is a non-interferometric technique for quantitative phase imaging.
    • Existing DPC methods can be limited by resolution and depth of field.
    • Quantitative phase imaging is crucial for label-free microscopy and biological sample analysis.

    Purpose of the Study:

    • To introduce and experimentally validate a novel pupil modulation differential phase contrast (PMDPC) imaging method.
    • To demonstrate the capability of PMDPC for high-resolution quantitative phase reconstruction.
    • To explore the digital extension of the depth of field in PMDPC.

    Main Methods:

    • Implemented PMDPC by filtering the Fourier domain of a sample with half-circle pupils.
    • Acquired phase gradient images using asymmetric pupil illumination.
    • Developed a reconstruction algorithm involving deconvolution of multiple phase gradient images.
    • Utilized a 4f system with a spatial light modulator (SLM) at the pupil plane for experimental realization.
    • Applied computational aberration removal and refocusing techniques.

    Main Results:

    • Achieved a quantitative phase image with a resolution of 1.73μm at a numerical aperture of 0.36.
    • Successfully extended the depth of field digitally by 20 times to ±50μm.
    • Maintained a high resolution of 1.76μm even with the extended depth of field.

    Conclusions:

    • PMDPC offers a robust and effective approach for high-resolution quantitative phase imaging.
    • The developed reconstruction algorithm and experimental setup enable precise phase measurements.
    • Digital refocusing and depth of field extension significantly enhance the versatility of PMDPC for various applications.