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Related Concept Videos

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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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An applied magnetic field causes loosely bound π-electrons in organic molecules to circulate, producing a local or induced diamagnetic field over a large spatial volume. As the molecules tumble in solution, the field generated by π-electrons in spherical substituents results in a zero net field. However, the net field generated by π-electrons in non-spherical substituents is not zero. The effect of this induced field depends on the orientation of the molecule with respect to B0,...
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Multiplexed barcoding image analysis has recently improved the characterization of the tumor microenvironment, permitting comprehensive studies of cell composition, functional state, and cell-cell interactions. Herein, we describe a staining and imaging protocol using the barcoding of oligonucleotide-conjugated antibodies and cycle imaging, which allows for the use of a high-dimensional image analysis...
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Related Experiment Video

Updated: Jan 19, 2026

Phase Contrast and Differential Interference Contrast DIC Microscopy
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Single-shot two-frame π-shifted spatially multiplexed interference phase microscopy.

Maciej Trusiak1, Jose-Angel Picazo-Bueno2, Krzysztof Patorski1

  • 1Warsaw University of Technology, Institute of Micromechanics and Photonics, Warsaw, Poland.

Journal of Biomedical Optics
|September 16, 2019
PubMed
Summary
This summary is machine-generated.

A new method, π-shifted spatially multiplexed interference microscopy (π-SMIM), offers fast and accurate quantitative phase imaging for challenging samples. This technique improves phase object analysis by reducing noise and enhancing contrast in microscopy images.

Keywords:
digital holographic microscopyfringe analysisinterference microscopyphase retrievalquantitative phase imaging

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Area of Science:

  • Microscopy
  • Optical Physics
  • Biomedical Imaging

Background:

  • Quantitative Phase Imaging (QPI) is crucial for analyzing transparent biological samples.
  • Traditional QPI methods can be slow, noisy, and suffer from low contrast.
  • Existing Spatially Multiplexed Interference Microscopy (SMIM) techniques have limitations.

Purpose of the Study:

  • To introduce an improved QPI technique, π-shifted SMIM (π-SMIM).
  • To enhance the speed, robustness, and accuracy of phase object analysis.
  • To enable in-vivo analysis of high dynamic range phase objects.

Main Methods:

  • Implemented single-shot, two-frame, π-shifted SMIM (π-SMIM) using a beam-splitter in a commercial microscope.
  • Simultaneously recorded two holograms with a π-radian phase shift.
  • Generated a π-hologram via subtractive superimposition for improved fringe modulation and reduced background.
  • Employed the Hilbert spiral transform for phase retrieval from the π-hologram.

Main Results:

  • Achieved high-speed (video frame rate) QPI capability.
  • Demonstrated superior phase retrieval quality compared to single hologram analysis.
  • Enabled accurate in-vivo analysis of dynamic phase objects.
  • Validated the technique using prostate cancer cells and flowing microbeads.

Conclusions:

  • π-SMIM provides a versatile, robust, fast, and accurate method for phase object analysis.
  • The technique overcomes limitations of previous SMIM implementations and traditional QPI.
  • π-SMIM facilitates advanced imaging of biological samples and dynamic processes.