High-Resolution Imaging and Analysis of Individual Nuclear Pore Complexes

Boris Fichtman; Saroj G Regmi; Mary Dasso; Amnon Harel

doi:10.1007/978-1-0716-2337-4_29

High-Resolution Imaging and Analysis of Individual Nuclear Pore Complexes

Methods Mol Biol. 2022:2502:461-471. doi: 10.1007/978-1-0716-2337-4_29.

Authors

Boris Fichtman¹, Saroj G Regmi², Mary Dasso², Amnon Harel³

Affiliations

¹ Azrieli Faculty of Medicine, Bar-Ilan University, Safed, Israel.
² Division of Molecular and Cellular Biology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA.
³ Azrieli Faculty of Medicine, Bar-Ilan University, Safed, Israel. amnon.harel@biu.ac.il.

PMID: 35412256
DOI: 10.1007/978-1-0716-2337-4_29

Abstract

Field emission scanning electron microscopy (FESEM) is a well-established technique for acquiring three-dimensional surface images of nuclear pore complexes (NPCs). We present an optimized protocol for the exposure of mammalian cell nuclei and direct surface imaging of nuclear envelopes by FESEM, allowing for a detailed morphological comparison of individual NPCs, without the need for averaging techniques. This provides a unique high resolution tool for studying the effects of cellular stress, specific genetic manipulations and inherited diseases on the ultrastructure of human NPCs.

Keywords: Auxin-inducible degron; Central channel particles; Mammalian cell culture; Nuclear envelope; Nuclear pore complex; Scanning electron microscopy.

Publication types

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

MeSH terms

Animals
Cell Nucleus / ultrastructure
Humans
Imaging, Three-Dimensional
Mammals
Microscopy, Electron, Scanning
Nuclear Envelope* / ultrastructure
Nuclear Pore* / metabolism