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Related Concept Videos

Phase Contrast and Differential Interference Contrast Microscopy01:26

Phase Contrast and Differential Interference Contrast Microscopy

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...
Confocal Fluorescence Microscopy01:16

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Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
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Super-resolution Fluorescence Microscopy

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 developed.

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Related Experiment Video

Updated: Jun 9, 2026

Quantitative Optical Microscopy: Measurement of Cellular Biophysical Features with a Standard Optical Microscope
14:09

Quantitative Optical Microscopy: Measurement of Cellular Biophysical Features with a Standard Optical Microscope

Published on: April 7, 2014

Surface-profile reconstruction using reflection differential phase-contrast microscopy.

M R Atkinson, A E Dixon, S Damaskinos

    Applied Optics
    |August 25, 2010
    PubMed
    Summary
    This summary is machine-generated.

    Optical differential phase-contrast microscopy accurately measures surface profiles. Feature height accuracy depends on steepness, with thin features measured directly and thicker ones requiring sample tilting.

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    Phase Contrast and Differential Interference Contrast (DIC) Microscopy
    06:49

    Phase Contrast and Differential Interference Contrast (DIC) Microscopy

    Published on: August 6, 2008

    Area of Science:

    • Physics
    • Materials Science
    • Microscopy

    Background:

    • Optical microscopy is crucial for analyzing sample surfaces.
    • Differential phase-contrast microscopy offers enhanced surface detail.
    • Accurate surface profiling is essential in various scientific fields.

    Purpose of the Study:

    • To outline the use of optical differential phase-contrast microscopy for surface profiling.
    • To determine the accuracy of feature height measurements based on feature geometry.
    • To investigate methods for measuring thicker samples.

    Main Methods:

    • Utilizing optical differential phase-contrast microscopy.
    • Calculating the range of accurate feature height determination as a function of feature side steepness.
    • Experimentally determining heights of thin features (<0.1 microm).
    • Employing sample tilting and oblique stage scanning for thicker samples.
    • Measuring reconstructed profile heights as a function of defocus.

    Main Results:

    • Accurate determination of thin feature heights (<0.1 microm) was achieved experimentally.
    • The accuracy of height determination is dependent on the steepness of the feature's side.
    • Techniques like sample tilting and oblique scanning are necessary for measuring thicker samples.
    • Reconstructed profile heights were successfully correlated with defocus levels.

    Conclusions:

    • Optical differential phase-contrast microscopy is a viable technique for surface profiling.
    • The method's accuracy is influenced by sample topography (feature steepness).
    • Specific experimental strategies are required to overcome limitations for thicker or steeper features.