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Related Concept Videos

Two-Dimensional Microscopy in Microbiology01:29

Two-Dimensional Microscopy in Microbiology

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Two-dimensional (2D) microscopy encompasses a range of optical techniques that capture images within a single focal plane, offering detailed representations of microscopic structures. These techniques are essential in biological and medical research, enabling the visualization of cellular and subcellular structures with different levels of contrast and specificity.There are several major types of 2D microscopy, each with strengths and applications.Bright-Field MicroscopyBright-field microscopy...
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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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Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

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A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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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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Three-Dimensional Microscopy in Microbiology01:28

Three-Dimensional Microscopy in Microbiology

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Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...
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Super-resolution Fluorescence Microscopy01:37

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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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Updated: Apr 10, 2026

Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions
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Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions

Published on: June 24, 2013

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Correlative microscopy.

Céline Loussert Fonta1, Bruno M Humbel1

  • 1Electron Microscopy Facility, University of Lausanne, Biophore, 1015 Lausanne, Switzerland.

Archives of Biochemistry and Biophysics
|June 15, 2015
PubMed
Summary
This summary is machine-generated.

Correlative microscopy combines light and electron imaging for detailed biomedical research. This powerful approach overcomes individual technique limitations, enabling sub-micrometer resolution of cellular structures and dynamics.

Keywords:
Correlative microscopyCryo-fixation and Freeze-substitutionEpoxyGFP green fluorescent proteinImmuno-(gold) labellingLight and Electron microscopyMethacrylateMouse brainSTEM scanning transmission electron microscopyTokuyasu cryo-sectioning

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Area of Science:

  • Biomedical Research
  • Microscopy Techniques

Background:

  • Correlative microscopy integrates diverse imaging modalities (light, electron, X-ray, NMR) for advanced biomedical research.
  • Light and electron microscopy combinations are increasingly vital, leveraging high resolution and live-cell imaging capabilities.

Purpose of the Study:

  • To review successful protocols for correlative light and electron microscopy (CLEM).
  • To identify common features in preparation steps for improved understanding.
  • To discuss new instruments and software facilitating CLEM.

Main Methods:

  • Combining light microscopy (LM) for live imaging and large field-of-view with electron microscopy (EM) for high-resolution ultrastructure.
  • Utilizing fluorescently tagged molecules or organelles in LM, followed by fixation and EM analysis of the same region.
  • Employing array tomography for 3D reconstruction of protein distribution by correlating multi-labeled sections with ultrastructure.

Main Results:

  • CLEM enables dissection of biological events at sub-micrometer resolution.
  • LM identifies specific cells or labeled proteins; EM reveals ultrastructural context.
  • Successful protocols are being developed to preserve fluorescence during EM preparation or convert labels for electron density.

Conclusions:

  • Correlative microscopy, particularly LM-EM, is a powerful tool for detailed biomedical investigations.
  • Optimized preparation protocols are crucial for successful CLEM.
  • Advancements in instrumentation and software are enhancing the accessibility and application of CLEM.