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Related Concept Videos

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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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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 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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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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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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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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Updated: Apr 23, 2026

Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions
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Correlative fluorescence and electron microscopy.

Randall T Schirra1, Peijun Zhang

  • 1Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.

Current Protocols in Cytometry
|October 2, 2014
PubMed
Summary
This summary is machine-generated.

Correlative fluorescence and electron microscopy (CFEM) integrates dynamic and ultrastructural data for deeper biological insights. This powerful technique enhances cellular component analysis and streamlines research, saving time and resources.

Keywords:
cell biologycorrelativecryoEMelectron microscopyfluorescenceimagingimmunolabelintegrated microscopelight microscopytomography

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Area of Science:

  • Cell Biology
  • Microscopy Techniques

Background:

  • Correlative fluorescence and electron microscopy (CFEM) combines dynamic imaging with ultrastructural details.
  • It offers a multimodal approach to understanding cellular components and their spatiotemporal dynamics.

Purpose of the Study:

  • To review the basic techniques and tools for utilizing CFEM.
  • To highlight the advantages of CFEM in biological research.

Main Methods:

  • Integration of fluorescence microscopy (dynamic, localization) and electron microscopy (ultrastructural).
  • Discussion of protocols and instrumentation for CFEM.

Main Results:

  • CFEM provides invaluable consensus between microscopy types, extending individual capabilities.
  • It reduces time and expense compared to separate analyses.

Conclusions:

  • CFEM is a powerful and increasingly adopted tool in cell biology.
  • Advancements in protocols and instruments are driving CFEM towards greater integration.