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Phase calibration target for quantitative phase imaging with ptychography.

T M Godden, A Muñiz-Piniella, J D Claverley

    Optics Express
    |May 4, 2016
    PubMed
    Summary
    This summary is machine-generated.

    A new standardized phase resolution target and protocol were developed to assess quantitative phase imaging (QPI) systems. This method ensures accurate measurements of optical path differences for transparent biological samples.

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    Area of Science:

    • Optical microscopy
    • Quantitative phase imaging
    • Biophysics

    Background:

    • Quantitative phase imaging (QPI) provides contrast for transparent objects by measuring refractive index and thickness variations.
    • QPI is crucial in quantitative biology for cell segmentation and calculating properties like dry mass and proliferation rate.
    • A lack of standardized methods for characterizing QPI system performance hinders reliable measurements.

    Purpose of the Study:

    • To present a standardized phase resolution target and measurement protocol for evaluating phase imaging systems.
    • To determine the transfer of spatial frequencies and sensitivity of phase imaging systems.
    • To address the need for reliable performance characterization in quantitative phase imaging.

    Main Methods:

    • Development of a phase resolution target with well-defined depth variations across a broad range of spatial frequencies.
    • Implementation of a measurement protocol to assess spatial frequency transfer and system sensitivity.
    • Quantitative phase imaging measurements using a ptychographic microscope, validated against Atomic Force Microscopy (AFM) and coherence scanning interferometry.

    Main Results:

    • The developed target and protocol enable the characterization of phase imaging system performance.
    • Ptychographic microscopy demonstrated quantitative nanometer sensitivity in optical path differences.
    • Accurate measurements were achieved for feature sizes ranging from micrometers to hundreds of micrometers.

    Conclusions:

    • The proposed phase resolution target and protocol offer a standardized approach for QPI system evaluation.
    • This standardization is essential for advancing quantitative biology applications relying on accurate phase measurements.
    • Ptychographic microscopy, when characterized with this method, provides highly sensitive and quantitative phase imaging.