High-Resolution scanning electron microscopy for the imaging of nuclear pore complexes and ran-mediated transport

Lihi Shaulov, Boris Fichtman, Amnon Harel

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

6 Scopus citations

Abstract

High-resolution scanning electron microscopy provides three-dimensional surface images of nuclear pore complexes (NPCs) embedded in the nuclear envelope. Here, we describe a method for exposing the nuclear surface in mammalian tissue culture cells for imaging by scanning electron microscopy. Hypotonic treatment is followed by low-speed centrifugation onto polylysine-coated silicon chips, without the use of detergents. This helps to preserve NPCs close to their native morphology, embedded in undamaged nuclear membranes. This method is particularly advantageous for combining high-resolution imaging of NPCs with mammalian genetic systems.

Original languageEnglish
Title of host publicationRas Signaling
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages253-261
Number of pages9
ISBN (Print)9781627037907
DOIs
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
Volume1120
ISSN (Print)1064-3745

Keywords

  • Mammalian cell culture
  • Nuclear envelope
  • Nuclear pore complex
  • Scanning electron microscopy

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