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Related Experiment Video

Updated: Jul 6, 2026

Single-Molecule Imaging of Nuclear Transport
12:13

Single-Molecule Imaging of Nuclear Transport

Published on: June 9, 2010

4Pi microscopy of the nuclear pore complex.

Jana Hüve1, Ramona Wesselmann, Martin Kahms

  • 1Institute of Medical Physics and Biophysics, and Center for Nanotechnology (CeNTech), University of Münster, Münster, Germany.

Biophysical Journal
|April 1, 2008
PubMed
Summary

Super-resolution microscopy precisely maps protein complexes like the nuclear pore complex (NPC). This advanced technique reveals cellular nanomachinery details with high accuracy.

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Proteins targeted to the nucleus carry nuclear localization signals or NLS recognized by import receptors in the cytosol. Similarly, proteins with nuclear export signals are recognized by export receptors. Import and export receptors are...

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Area of Science:

  • Cell Biology
  • Biophysics
  • Microscopy

Background:

  • Understanding the structure of cellular protein complexes is crucial for cell biology.
  • Existing microscopy techniques have limitations in resolving nanoscale features of protein complexes.

Purpose of the Study:

  • To evaluate the capability of super-resolution fluorescence microscopy in resolving topographic features of single cellular protein complexes.
  • To investigate the structural organization of the nuclear pore complex (NPC).

Main Methods:

  • Utilized a two-photon 4Pi microscope with high axial resolution (110-130 nm) and localization accuracy (5-10 nm).
  • Employed immune-labeling techniques on HeLa cells and cells expressing green fluorescent protein constructs.
  • Compared 4Pi microscopy with conventional confocal microscopy.

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Examination of Mitotic and Meiotic Fission Yeast Nuclear Dynamics by Fluorescence Live-cell Microscopy
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Examination of Mitotic and Meiotic Fission Yeast Nuclear Dynamics by Fluorescence Live-cell Microscopy

Published on: June 24, 2019

Related Experiment Videos

Last Updated: Jul 6, 2026

Single-Molecule Imaging of Nuclear Transport
12:13

Single-Molecule Imaging of Nuclear Transport

Published on: June 9, 2010

Examination of Mitotic and Meiotic Fission Yeast Nuclear Dynamics by Fluorescence Live-cell Microscopy
12:04

Examination of Mitotic and Meiotic Fission Yeast Nuclear Dynamics by Fluorescence Live-cell Microscopy

Published on: June 24, 2019

Main Results:

  • 4Pi microscopy provided significantly better resolution of NPCs compared to confocal microscopy.
  • Successfully resolved and measured distances between specific epitopes within single NPCs (e.g., 152 ± 30 nm).
  • Determined distances from the NPC center to different structural components, aligning with previous estimates.

Conclusions:

  • Super-resolution fluorescence microscopy is a powerful tool for analyzing individual protein complexes.
  • This method enables detailed analysis of cellular nanomachinery.
  • The study validates the utility of 4Pi microscopy for structural investigations at the nanoscale.