Correlative super-resolution fluorescence and electron microscopy of the nuclear pore complex with molecular resolution

Anna Löschberger, Christian Franke, Georg Krohne, Sebastian van de Linde, Markus Sauer

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

Here, we combine super-resolution fluorescence localization microscopy with scanning electron microscopy to map the position of proteins of nuclear pore complexes in isolated Xenopus laevis oocyte nuclear envelopes with molecular resolution in both imaging modes. We use the periodic molecular structure of the nuclear pore complex to superimpose direct stochastic optical reconstruction microscopy images with a precision of <20 nm on electron micrographs. The correlative images demonstrate quantitative molecular labeling and localization of nuclear pore complex proteins by standard immunocytochemistry with primary and secondary antibodies and reveal that the nuclear pore complex is composed of eight gp210 (also known as NUP210) protein homodimers. In addition, we find subpopulations of nuclear pore complexes with ninefold symmetry, which are found occasionally among the more typical eightfold symmetrical structures.

LanguageEnglish
Pages4351-4355
Number of pages5
JournalJournal of Cell Science
Volume127
Issue number20
Early online date21 Aug 2014
DOIs
Publication statusPublished - 14 Oct 2014
Externally publishedYes

Fingerprint

Nuclear Pore
Fluorescence Microscopy
Nuclear Pore Complex Proteins
Electron Microscopy
Computer-Assisted Image Processing
Nuclear Envelope
Xenopus laevis
Molecular Structure
Electron Scanning Microscopy
Oocytes
Microscopy
Immunohistochemistry
Electrons
Antibodies
Proteins

Keywords

  • correlative electron and super-resolution fluorescence microscopy
  • localization microscopy
  • nuclear pore complex
  • dSTORM
  • immunocytochemistry

Cite this

Löschberger, Anna ; Franke, Christian ; Krohne, Georg ; van de Linde, Sebastian ; Sauer, Markus. / Correlative super-resolution fluorescence and electron microscopy of the nuclear pore complex with molecular resolution. In: Journal of Cell Science. 2014 ; Vol. 127, No. 20. pp. 4351-4355.
@article{16fce366e2304128ad30f2fcdce554d0,
title = "Correlative super-resolution fluorescence and electron microscopy of the nuclear pore complex with molecular resolution",
abstract = "Here, we combine super-resolution fluorescence localization microscopy with scanning electron microscopy to map the position of proteins of nuclear pore complexes in isolated Xenopus laevis oocyte nuclear envelopes with molecular resolution in both imaging modes. We use the periodic molecular structure of the nuclear pore complex to superimpose direct stochastic optical reconstruction microscopy images with a precision of <20 nm on electron micrographs. The correlative images demonstrate quantitative molecular labeling and localization of nuclear pore complex proteins by standard immunocytochemistry with primary and secondary antibodies and reveal that the nuclear pore complex is composed of eight gp210 (also known as NUP210) protein homodimers. In addition, we find subpopulations of nuclear pore complexes with ninefold symmetry, which are found occasionally among the more typical eightfold symmetrical structures.",
keywords = "correlative electron and super-resolution fluorescence microscopy, localization microscopy, nuclear pore complex, dSTORM, immunocytochemistry",
author = "Anna L{\"o}schberger and Christian Franke and Georg Krohne and {van de Linde}, Sebastian and Markus Sauer",
note = "{\circledC} 2014. Published by The Company of Biologists Ltd.",
year = "2014",
month = "10",
day = "14",
doi = "10.1242/jcs.156620",
language = "English",
volume = "127",
pages = "4351--4355",
journal = "Journal of Cell Science",
issn = "0021-9533",
number = "20",

}

Correlative super-resolution fluorescence and electron microscopy of the nuclear pore complex with molecular resolution. / Löschberger, Anna; Franke, Christian; Krohne, Georg; van de Linde, Sebastian; Sauer, Markus.

In: Journal of Cell Science, Vol. 127, No. 20, 14.10.2014, p. 4351-4355.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Correlative super-resolution fluorescence and electron microscopy of the nuclear pore complex with molecular resolution

AU - Löschberger, Anna

AU - Franke, Christian

AU - Krohne, Georg

AU - van de Linde, Sebastian

AU - Sauer, Markus

N1 - © 2014. Published by The Company of Biologists Ltd.

PY - 2014/10/14

Y1 - 2014/10/14

N2 - Here, we combine super-resolution fluorescence localization microscopy with scanning electron microscopy to map the position of proteins of nuclear pore complexes in isolated Xenopus laevis oocyte nuclear envelopes with molecular resolution in both imaging modes. We use the periodic molecular structure of the nuclear pore complex to superimpose direct stochastic optical reconstruction microscopy images with a precision of <20 nm on electron micrographs. The correlative images demonstrate quantitative molecular labeling and localization of nuclear pore complex proteins by standard immunocytochemistry with primary and secondary antibodies and reveal that the nuclear pore complex is composed of eight gp210 (also known as NUP210) protein homodimers. In addition, we find subpopulations of nuclear pore complexes with ninefold symmetry, which are found occasionally among the more typical eightfold symmetrical structures.

AB - Here, we combine super-resolution fluorescence localization microscopy with scanning electron microscopy to map the position of proteins of nuclear pore complexes in isolated Xenopus laevis oocyte nuclear envelopes with molecular resolution in both imaging modes. We use the periodic molecular structure of the nuclear pore complex to superimpose direct stochastic optical reconstruction microscopy images with a precision of <20 nm on electron micrographs. The correlative images demonstrate quantitative molecular labeling and localization of nuclear pore complex proteins by standard immunocytochemistry with primary and secondary antibodies and reveal that the nuclear pore complex is composed of eight gp210 (also known as NUP210) protein homodimers. In addition, we find subpopulations of nuclear pore complexes with ninefold symmetry, which are found occasionally among the more typical eightfold symmetrical structures.

KW - correlative electron and super-resolution fluorescence microscopy

KW - localization microscopy

KW - nuclear pore complex

KW - dSTORM

KW - immunocytochemistry

UR - http://jcs.biologists.org/

U2 - 10.1242/jcs.156620

DO - 10.1242/jcs.156620

M3 - Article

VL - 127

SP - 4351

EP - 4355

JO - Journal of Cell Science

T2 - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 20

ER -