Imaging metazoan nuclear pore complexes by field emission scanning electron microscopy

Boris Fichtman, Lihi Shaulov, Amnon Harel

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

16 Scopus citations

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.

Original languageEnglish
Title of host publicationMethods in Cell Biology
PublisherAcademic Press Inc.
Pages41-58
Number of pages18
DOIs
StatePublished - 2014

Publication series

NameMethods in Cell Biology
Volume122
ISSN (Print)0091-679X

Bibliographical note

Funding Information:
The authors thank Eugenia Klein and Michael Elbaum for advice on FESEM analysis, Ralph Neujahr and Xiong Liu (Carl Zeiss group, Germany) for their assistance in obtaining the images in Fig. 2.5 and Nitzan Shabtay for assistance with Fig. 2.4 . This work was supported by a research grant from the Israel Science Foundation (1072/10) to A.H.

Keywords

  • Anchored nuclei
  • Iridium coating
  • Nuclear envelope
  • Nuclear pore complex
  • Scanning electron microscopy

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