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Related Concept Videos

Overview of Electron Microscopy01:25

Overview of Electron Microscopy

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.
Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

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...
Transmission Electron Microscopy01:15

Transmission Electron Microscopy

In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light in an optical microscope—developed the first prototype of the electron microscope. This development led to the development of the field of electron microscopy. In the transmission electron microscope (TEM), electrons are produced by a hot tungsten element and accelerated by a potential difference in an electron gun, which gives them up to 400 keV in...
Scanning Electron Microscopy01:07

Scanning Electron Microscopy

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.
Fundamental Principles
Accelerated...
Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

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...
Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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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Single Particle Electron Microscopy Reconstruction of the Exosome Complex Using the Random Conical Tilt Method
12:10

Single Particle Electron Microscopy Reconstruction of the Exosome Complex Using the Random Conical Tilt Method

Published on: March 28, 2011

Single particle electron microscopy.

Wilson C Y Lau1, John L Rubinstein

  • 1Molecular Structure and Function Program, Departments of Biochemistry and Medical Biophysics, The Hospital for Sick Children, The University of Toronto, Toronto, ON, Canada.

Methods in Molecular Biology (Clifton, N.J.)
|November 8, 2012
PubMed
Summary
This summary is machine-generated.

Single particle electron microscopy (SPEM) provides near-native structural analysis of protein complexes. This guide offers practical steps for initiating SPEM studies, covering specimen preparation, imaging, and data analysis strategies.

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Last Updated: May 17, 2026

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Published on: March 28, 2011

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Area of Science:

  • Biophysics
  • Structural Biology
  • Biochemistry

Background:

  • Single particle electron microscopy (SPEM) is a powerful technique for determining protein complex structures under near-native conditions.
  • Significant advancements in specimen preparation, imaging, and data processing have been achieved, yet the field continues to evolve.

Purpose of the Study:

  • To provide practical guidelines for researchers beginning single particle electron microscopy studies.
  • To outline essential steps for specimen preparation, imaging parameter selection, and appropriate image analysis method selection for diverse samples.

Main Methods:

  • Specimen preparation techniques for cryo-electron microscopy.
  • Selection of optimal imaging conditions, including defocus and electron dose.
  • Overview of various image processing and analysis workflows for single particle data.

Main Results:

  • The chapter details a systematic approach to initiating SPEM projects.
  • It emphasizes the importance of tailored strategies for specimen handling, data acquisition, and computational analysis based on sample characteristics.

Conclusions:

  • This chapter serves as a foundational resource for researchers new to single particle electron microscopy.
  • It empowers scientists to effectively apply SPEM for structural elucidation of biological macromolecules.