Imaging metazoan nuclear pore complexes by field emission scanning electron microscopy

Methods Cell Biol. 2014;122:41-58. doi: 10.1016/B978-0-12-417160-2.00002-3.

Abstract

High resolution three-dimensional surface images of nuclear pore complexes (NPCs) can be obtained by field emission scanning electron microscopy. We present a short retrospective view starting from the early roots of microscopy, through the discovery of the cell nucleus and the development of some modern techniques for sample preparation and imaging. Detailed protocols are presented for assembling anchored nuclei in a Xenopus cell-free reconstitution system and for the exposure of the nuclear surface in mammalian cell nuclei. Immunogold labeling of metazoan NPCs and a promising new technique for delicate coating with iridium are also discussed.

Keywords: Anchored nuclei; Iridium coating; Nuclear envelope; Nuclear pore complex; Scanning electron microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell-Free System
  • Fixatives
  • Imaging, Three-Dimensional
  • Iridium / chemistry
  • Microscopy, Electron, Scanning / methods*
  • Nuclear Pore / physiology*
  • Oocytes / cytology*
  • Staining and Labeling / methods
  • Tissue Fixation / methods
  • Xenopus laevis

Substances

  • Fixatives
  • Iridium