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Related Concept Videos

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
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Preparation of Samples for Electron Microscopy01:20

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To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
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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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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.
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...
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Related Experiment Video

Updated: Jul 14, 2025

Cell Culture on Silicon Nitride Membranes and Cryopreparation for Synchrotron X-ray Fluorescence Nano-analysis
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Advancement in Cellular Topographic and Nanoparticle Capture Imaging by High Resolution Microscopy Incorporating a

Kunihiro Uryu1,2, Nadine Soplop1, Timothy P Sheahan3

  • 1Electron Microscopy Resource Center, The Rockefeller University, New York, NY 10065, USA.

Biorxiv : the Preprint Server for Biology
|October 9, 2023
PubMed
Summary

A new freeze-drying method with gaseous nitrogen (FDGN) preserves cell structure and fluorescence for high-resolution scanning electron microscopy (SEM). This advance enables detailed visualization of extracellular vesicles and host cell interactions.

Keywords:
correlative light and electron microscopygaseous nitrogenhelium ion microscopyhepatitis C virusscanning electron microscopytransmission electron microscopytumor-derived extracellular vesicles

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Plunge Freezing: A Tool for the Ultrastructural and Immunolocalization Studies of Suspension Cells in Transmission Electron Microscopy
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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Biotechnology

Background:

  • Conventional scanning electron microscopy (SEM) preparation using osmium tetroxide (OsO4) and ethanol limits resolution and compromises fluorescent signals.
  • This hinders correlative light and electron microscopy (CLEM) for studying cellular structures and fluorescent reporter proteins.

Approach:

  • Developed a novel freeze-drying method with gaseous nitrogen (FDGN) to overcome limitations of conventional SEM preparation.
  • FDGN processing preserves cellular cyto-architecture and retains fluorescence signals for high-resolution imaging.

Key Points:

  • FDGN enables visualization of detailed membrane topography and fluorescent reporter proteins simultaneously.
  • The method is compatible with various high-resolution imaging systems for data validation.
  • Demonstrated detailed insight into viral or tumor-derived extracellular vesicle (TEV)-host cell interactions.

Conclusions:

  • FDGN is a valuable technique for high-resolution SEM and CLEM, overcoming previous technological barriers.
  • This method facilitates the study of extracellular vesicle-host interactions.
  • Potential applications include developing new antiviral strategies and therapeutic uses for TEVs.