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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...
Scanning Electron Microscopy01:07

Scanning Electron Microscopy

A scanning electron microscope (SEM) is used to study the surface features of a sample by using an electron beam that scans the sample surface in a two-dimensional manner. Typically, areas between ~1 centimeter to 5 micrometers in width can be imaged. SEM can be used to image bacteria, viruses, tissues as well as larger samples like insects. Conventional SEM gives a magnification ranging from 20X to 30,000X and spatial resolution of 50 to 100 nanometers.
Fundamental Principles
Accelerated...
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

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,...
Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
X-ray Imaging01:24

X-ray Imaging

German physicist Wilhelm Röntgen (1845–1923) was experimenting with electrical current when he discovered that a mysterious and invisible "ray" would pass through his flesh but leave an outline of his bones on a screen coated with a metal compound. In 1895, Röntgen made the first durable record of the internal parts of a living human: an "X-ray" image (as it came to be called) of his wife’s hand. Scientists worldwide quickly began their own experiments with X-rays, and by 1900, X-ray was widely...
X-ray Diffraction of Biological Samples01:10

X-ray Diffraction of Biological Samples

X-ray diffraction or XRD is an analytical tool that utilizes X-rays to study ordered structures such as crystalline organic and inorganic samples, polycrystalline materials, proteins, carbohydrates, and drugs.
According to Bragg's law, when X-rays strike the sample positioned on a stage, the rays are  scattered by the electron clouds around the sample atoms. The  X-ray diffraction or scattering is caused by constructive interference of the X-ray waves that reflect off the internal crystal...

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Updated: Jun 2, 2026

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

Phase Contrast and Differential Interference Contrast (DIC) Microscopy

Published on: August 6, 2008

Zernike phase contrast in scanning microscopy with X-rays.

Christian Holzner1, Michael Feser, Stefan Vogt

  • 1Department of Physics and Astronomy, Stony Brook University, Nicolls Road, Stony Brook, New York 11794, USA.

Nature Physics
|May 6, 2011
PubMed
Summary

Researchers integrated Zernike phase contrast into scanning X-ray microscopy. This technique reveals structural details alongside elemental mapping, enhancing analysis of biological specimens.

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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

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Last Updated: Jun 2, 2026

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

Phase Contrast and Differential Interference Contrast (DIC) Microscopy

Published on: August 6, 2008

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

Area of Science:

  • X-ray microscopy
  • Phase contrast imaging
  • Materials science

Background:

  • Scanning X-ray microscopy (SXM) provides elemental mapping via X-ray fluorescence but lacks structural information.
  • Full-field X-ray microscopes utilize Zernike phase contrast for visualizing weakly absorbing samples.
  • Coupling structural and elemental data in SXM is crucial for enhanced analysis.

Purpose of the Study:

  • To implement Zernike phase contrast in scanning X-ray microscopy.
  • To enable simultaneous acquisition of structural and elemental data.
  • To demonstrate the benefits of correlating morphology with elemental composition.

Main Methods:

  • Adaptation of Zernike phase contrast principles for SXM based on reciprocity.
  • Raster scanning of the sample with a focused X-ray beam.
  • Simultaneous collection of transmission (phase contrast) and fluorescence (elemental) signals.

Main Results:

  • Successful implementation of Zernike phase contrast in SXM.
  • Acquisition of high-resolution images showing both structural detail and elemental distribution.
  • Demonstration of complementary information obtained from combined techniques.

Conclusions:

  • Zernike phase contrast is effectively integrated into SXM.
  • The method enhances the analysis of weakly absorbing samples by providing simultaneous structural and elemental information.
  • Correlating morphology with elemental composition in biological specimens is significantly improved.