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Related Concept Videos

Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

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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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Three-Dimensional Microscopy in Microbiology01:28

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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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Total Internal Reflection Fluorescence Microscopy01:05

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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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Electron Microscope Tomography and Single-particle Reconstruction01:07

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Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
Electron Tomography
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Immunofluorescence Microscopy01:12

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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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Updated: Mar 2, 2026

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

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A fully integrated, three-dimensional fluorescence to electron microscopy correlative workflow.

Claudia S López1, Cedric Bouchet-Marquis2, Christopher P Arthur3

  • 1Oregon Health and Sciences University, Portland, OR, United States.

Methods in Cell Biology
|May 23, 2017
PubMed
Summary
This summary is machine-generated.

Correlative light and electron microscopy (CLEM) overcomes fluorescence microscopy limitations. This study demonstrates an integrated CLEM workflow for enhanced cellular structure and protein localization analysis in breast cancer cells.

Keywords:
3D FIB-SEMCLEMCorrelative imaging techniqueElectron microscopyFluorescence microscopy

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Author Spotlight: Advancements in Correlative Light and Electron Microscopy with Fluorescent Protein Preservation
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Author Spotlight: Advancements in Correlative Light and Electron Microscopy with Fluorescent Protein Preservation
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Area of Science:

  • Cell Biology
  • Microscopy
  • Biotechnology

Background:

  • Fluorescence microscopy offers specific labeling but has resolution limits and poor contrast.
  • Studying cellular structure and protein localization is challenging with traditional microscopy.
  • Correlative Light and Electron Microscopy (CLEM) integrates multiple imaging modalities.

Purpose of the Study:

  • To demonstrate an integrated correlative light and electron microscopy (CLEM) workflow.
  • To showcase the FEI CorrSight with MAPS for reliable and rapid CLEM data correlation.
  • To apply the integrated CLEM workflow to fluorescently tagged MCF7 breast cancer cells.

Main Methods:

  • Utilizing an integrated CLEM workflow instrument (FEI CorrSight with MAPS).
  • Correlating three-dimensional time-lapse confocal fluorescence microscopy data.
  • Correlating three-dimensional focused ion beam-scanning electron microscopy data.

Main Results:

  • Demonstration of a complete, integrated CLEM workflow.
  • Successful correlation of fluorescence and electron microscopy data.
  • Application of the workflow to MCF7 breast cancer cells.

Conclusions:

  • Integrated CLEM workflows provide a reliable, reproducible, and faster approach for cellular imaging.
  • This method enhances the study of cellular structure and protein localization.
  • The demonstrated workflow is applicable to cancer cell research.