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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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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
Electron tomography can be performed either in TEM or STEM (scanning transmission...
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Updated: Aug 29, 2025

Single Particle Cryo-Electron Microscopy: From Sample to Structure
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Single-particle cryo-EM structures from iDPC-STEM at near-atomic resolution.

Ivan Lazić1, Maarten Wirix2, Max Leo Leidl3,4,5,6

  • 1Materials and Structural Analysis Division, Thermo Fisher Scientific, Eindhoven, Netherlands. ivan.lazic@thermofisher.com.

Nature Methods
|September 6, 2022
PubMed
Summary
This summary is machine-generated.

Scanning transmission electron microscopy (STEM) with integrated differential phase contrast (iDPC-STEM) successfully imaged biological samples. This method achieved high-resolution cryo-electron microscopy (cryo-EM) structures for keyhole limpet hemocyanin and tobacco mosaic virus.

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Area of Science:

  • Structural Biology
  • Microscopy Techniques
  • Biophysics

Background:

  • Electron cryomicroscopy (cryo-EM) is crucial for determining high-resolution 3D structures of biological macromolecules.
  • Conventional transmission electron microscopy (CTEM) with large underfocuses is the standard cryo-EM imaging method.
  • Limitations in contrast transfer can affect resolution in CTEM imaging.

Purpose of the Study:

  • To evaluate the applicability of scanning transmission electron microscopy (STEM) for cryo-EM structure determination.
  • To assess the performance of integrated differential phase contrast STEM (iDPC-STEM) on vitrified biological specimens.
  • To compare iDPC-STEM results with conventional CTEM data.

Main Methods:

  • Application of iDPC-STEM to cryo-EM test specimens: keyhole limpet hemocyanin (KLH) and tobacco mosaic virus (TMV).
  • Acquisition of micrographs with complete contrast transfer to high resolution.
  • Utilizing single-particle reconstruction methods for 3D structure determination.

Main Results:

  • Successful cryo-EM structure determination of KLH at 6.5 Å resolution using iDPC-STEM.
  • Achieved 3.5 Å resolution for TMV structure determination with iDPC-STEM.
  • Cryo-EM structures obtained via iDPC-STEM showed identical features to those from CTEM.

Conclusions:

  • STEM imaging, specifically iDPC-STEM, is a viable method for cryo-EM structure determination of biological macromolecules.
  • iDPC-STEM enables near-atomic resolution cryo-EM structure determination of vitrified single-particle specimens.
  • This technique offers a complementary approach to CTEM for high-resolution structural analysis.