title: Imaging metazoan nuclear pore complexes by field emission scanning electron microscopy.
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.
authors.affiliation: Faculty of Medicine in the Galilee, Bar-Ilan University, Safed, Israel.
authors.forename: Boris
authors.initials: B
authors.lastname: Fichtman
authors.affiliation: Faculty of Medicine in the Galilee, Bar-Ilan University, Safed, Israel.
authors.forename: Lihi
authors.initials: L
authors.lastname: Shaulov
authors.affiliation: Faculty of Medicine in the Galilee, Bar-Ilan University, Safed, Israel.
authors.forename: Amnon
authors.initials: A
authors.lastname: Harel
mesh_terms: D000818:Animals; D002474:Cell-Free System; D005404:Fixatives; D021621:Imaging, Three-Dimensional; D007495:Iridium / Q000737:chemistry; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D022022:Nuclear Pore / Q000502:physiology*; D009865:Oocytes / Q000166:cytology*; D013194:Staining and Labeling / Q000379:methods; D016707:Tissue Fixation / Q000379:methods; D014982:Xenopus laevis
chemical_list: D005404:Fixatives; D007495:Iridium
keywords: Anchored nuclei; Iridium coating; Nuclear envelope; Nuclear pore complex; Scanning electron microscopy