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Phase Contrast and Differential Interference Contrast Microscopy01:26

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In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...
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Quantitative Optical Microscopy: Measurement of Cellular Biophysical Features with a Standard Optical Microscope
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High-speed Fourier ptychographic microscopy for quantitative phase imaging.

Yi Xiao, Shiyuan Wei, Shaolong Xue

    Optics Letters
    |October 1, 2021
    PubMed
    Summary

    We developed a high-speed Fourier ptychographic microscopy (FPM) technique for real-time imaging. This method achieves over 42 frames per second, enabling nanometer-scale sensitivity for red blood cell imaging and subcellular resolution in cancer tissues.

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    Area of Science:

    • Computational imaging
    • Optical microscopy
    • Biomedical imaging

    Background:

    • Fourier ptychographic microscopy (FPM) offers super-resolution imaging beyond optical limits.
    • Current FPM techniques suffer from low imaging speed due to long acquisition times and weak illumination, hindering real-time applications.

    Purpose of the Study:

    • To develop a high-speed FPM method for real-time quantitative phase and intensity imaging.
    • To enhance the imaging speed of FPM beyond 42 frames per second (fps) while maintaining high resolution.

    Main Methods:

    • Utilized laser illumination for increased intensity.
    • Employed digital micro-mirror devices (DMDs) for rapid illumination angle scanning.
    • Implemented a novel FPM approach to achieve high-speed data acquisition and processing.

    Main Results:

    • Achieved quantitative phase and intensity imaging at over 42 fps with approximately 1 µm lateral resolution.
    • Demonstrated nanometer-scale sensitivity in detecting red blood cell membrane height fluctuations.
    • Resolved subcellular features in stained cancer tissue slices using intensity imaging.

    Conclusions:

    • The proposed high-speed FPM method significantly advances real-time imaging capabilities in microscopy.
    • This technique enables sensitive, high-resolution imaging of biological samples, including dynamic cellular processes and tissue morphology.