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Related Concept Videos

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Nuclear protein sorting is the selective trafficking of histones, polymerases, gene regulatory proteins into the nucleus and exporting RNAs and ribosomes to the cytosol. It is a tightly controlled process that regulates gene expression within a cell.
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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Related Experiment Video

Updated: Apr 21, 2026

Single-Molecule Imaging of Nuclear Transport
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4Pi microscopy of the nuclear pore complex.

Martin Kahms1, Jana Hüve, Reiner Peters

  • 1Institute for Medical Physics and Biophysics, University Muenster, Robert-Koch-Strasse 31, Muenster, 48149, USA.

Methods in Molecular Biology (Clifton, N.J.)
|November 14, 2014
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Summary

4Pi microscopy, a super-resolution technique, achieves 100 nm axial resolution by interfering light waves. This advanced fluorescence imaging method visualizes cellular structures like the nuclear pore complex with unprecedented detail.

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Cell Biology

Background:

  • 4Pi microscopy is a far-field fluorescence technique utilizing interfering light.
  • It offers significantly higher resolution compared to conventional confocal microscopy.

Purpose of the Study:

  • To describe the fundamental principles of 4Pi microscopy.
  • To illustrate its application in cell biology, focusing on the nuclear pore complex.

Main Methods:

  • Utilizes two opposing illuminating beams with adjusted wave fronts for constructive interference.
  • Analyzes the resulting point spread function with a 100 nm axial full width at half maximum.

Main Results:

  • Achieves an axial resolution 6-7 times better than confocal microscopy.
  • Demonstrates the capability of visualizing large protein assemblies, such as the nuclear pore complex.

Conclusions:

  • 4Pi microscopy provides enhanced resolution for detailed biological imaging.
  • It is a powerful tool for studying complex cellular structures in their native environment.