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Related Concept Videos

Phase Contrast and Differential Interference Contrast Microscopy01:26

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Phase-Contrast Microscopes
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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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Quantitative Optical Microscopy: Measurement of Cellular Biophysical Features with a Standard Optical Microscope
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Epi-illumination diffraction phase microscopy with white light.

C Edwards, B Bhaduri, B G Griffin

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    |November 1, 2014
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    Summary
    This summary is machine-generated.

    Researchers developed a new reflection-based white light diffraction phase microscope (epi-wDPM). This advanced imaging technique achieves sub-nanometer noise levels for opaque samples, offering high-resolution insights.

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    Area of Science:

    • Optical microscopy
    • Phase contrast imaging
    • Nanotechnology

    Background:

    • Diffraction phase microscopy (DPM) is a powerful technique for label-free imaging.
    • Existing DPM systems often face limitations in sample accessibility and illumination types.
    • Reflection-based microscopy is crucial for opaque and non-transparent samples.

    Purpose of the Study:

    • To introduce the first reflection-based epi-illumination white light diffraction phase microscope (epi-wDPM).
    • To demonstrate its capability for high-resolution imaging of opaque samples.
    • To showcase its performance using biological samples.

    Main Methods:

    • Development of a novel epi-wDPM system integrating off-axis, common-path, and white light principles in reflection geometry.
    • Utilizing single-shot acquisition for imaging opaque samples.
    • Verification through measurements of control samples with known dimensions and comparison with established techniques.

    Main Results:

    • The epi-wDPM system achieved sub-nanometer spatial and temporal noise levels.
    • Successful imaging of opaque samples with high resolution.
    • Demonstrated tradeoffs between low and high spatial coherence using gold-coated HeLa cells.

    Conclusions:

    • The developed epi-wDPM is a significant advancement in reflection-based phase microscopy.
    • It offers high-performance imaging capabilities for a range of opaque samples.
    • The system provides valuable insights into sample topography and optical path difference.