An introduction to sample preparation and imaging by cryo-electron microscopy for structural biology

Methods. 2016 May 1:100:3-15. doi: 10.1016/j.ymeth.2016.02.017. Epub 2016 Feb 28.

Abstract

Transmission electron microscopy (EM) is a versatile technique that can be used to image biological specimens ranging from intact eukaryotic cells to individual proteins >150kDa. There are several strategies for preparing samples for imaging by EM, including negative staining and cryogenic freezing. In the last few years, cryo-EM has undergone a 'resolution revolution', owing to both advances in imaging hardware, image processing software, and improvements in sample preparation, leading to growing number of researchers using cryo-EM as a research tool. However, cryo-EM is still a rapidly growing field, with unique challenges. Here, we summarise considerations for imaging of a range of specimens from macromolecular complexes to cells using EM.

Keywords: Electron microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Cryoelectron Microscopy / methods*
  • Freezing
  • Humans
  • Imaging, Three-Dimensional
  • Macromolecular Substances / chemistry
  • Macromolecular Substances / ultrastructure
  • Microscopy, Electron, Transmission
  • Models, Molecular
  • Vitrification

Substances

  • Macromolecular Substances