Biological applications of phase-contrast electron microscopy

Methods Mol Biol. 2014:1117:385-99. doi: 10.1007/978-1-62703-776-1_18.

Abstract

Here, I review the principles and applications of phase-contrast electron microscopy using phase plates. First, I develop the principle of phase contrast based on a minimal model of microscopy, introducing a double Fourier-transform process to mathematically formulate the image formation. Next, I explain four phase-contrast (PC) schemes, defocus PC, Zernike PC, Hilbert differential contrast, and schlieren optics, as image-filtering processes in the context of the minimal model, with particular emphases on the Zernike PC and corresponding Zernike phase plates. Finally, I review applications of Zernike PC cryo-electron microscopy to biological systems such as protein molecules, virus particles, and cells, including single-particle analysis to delineate three-dimensional (3D) structures of protein and virus particles and cryo-electron tomography to reconstruct 3D images of complex protein systems and cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Cryoelectron Microscopy / methods*
  • Microscopy, Phase-Contrast / methods*