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Related Concept Videos

Atomic Force Microscopy01:08

Atomic Force Microscopy

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Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
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Overview of Electron Microscopy01:25

Overview of Electron Microscopy

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The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.
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Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

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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.
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Atomic Fluorescence Spectroscopy01:29

Atomic Fluorescence Spectroscopy

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Atomic fluorescence spectroscopy (AFS) is an analytical technique that involves the electronic transitions of atoms in a flame, furnace, or plasma being excited by electromagnetic (EM) radiation. When these atoms absorb energy, they become excited and subsequently release energy as they return to their original state. This emitted light, or "fluorescence," is observed at a right angle to the incident beam. Both absorption and emission processes transpire at distinct wavelengths, which...
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Scanning Electron Microscopy01:07

Scanning Electron Microscopy

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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.
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Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

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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...
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Author Spotlight: Advancements in Correlative Light and Electron Microscopy with Fluorescent Protein Preservation
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CLAFEM: Correlative light atomic force electron microscopy.

Sébastien Janel1, Elisabeth Werkmeister1, Antonino Bongiovanni1

  • 1Univ. Lille, CNRS UMR 8204, Inserm U1019, CHU Lille, Institut Pasteur de Lille - CIIL - Center for Infection and Immunity of Lille, Lille, France.

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

Correlative Light Atomic Force Electron Microscopy (CLAFEM) integrates multiple imaging techniques. This powerful approach provides nanoscale topographical and biomechanical data for biological samples, revealing cellular details.

Keywords:
Atomic force microscopyCell elasticityCorrelative microscopyScanning electron microscopySuperresolution fluorescence microscopyTransmission electron microscopy

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Area of Science:

  • Biophysics
  • Cell Biology
  • Microscopy

Background:

  • Atomic Force Microscopy (AFM) offers precise topographical and biomechanical data for biological samples.
  • Correlative microscopy techniques are essential for multi-scale analysis of cellular structures and functions.

Purpose of the Study:

  • To introduce and validate Correlative Light Atomic Force Electron Microscopy (CLAFEM).
  • To demonstrate CLAFEM's capability in analyzing cellular processes like bacterial infection and cytoskeleton dynamics.

Main Methods:

  • Integration of AFM with super-resolution light microscopy and electron microscopy (transmission or scanning).
  • Application of CLAFEM on both fixed and living cells.

Main Results:

  • CLAFEM provides complementary nanoscale topographical and piconewton-scale biomechanical (elasticity) data.
  • The technique enables detailed ultrastructural analysis alongside functional and compositional information.

Conclusions:

  • CLAFEM is a versatile correlative technique for comprehensive cellular analysis.
  • This method enhances understanding of cellular mechanisms by combining diverse imaging modalities.