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Related Experiment Video

Updated: Apr 19, 2026

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Live-cell multiplane three-dimensional super-resolution optical fluctuation imaging.

Stefan Geissbuehler1, Azat Sharipov1, Aurélien Godinat2

  • 1cole Polytechnique Fédérale de Lausanne, Laboratoire d'Optique Biomédicale, 1015 Lausanne, Switzerland.

Nature Communications
|December 19, 2014
PubMed
Summary
This summary is machine-generated.

Super-resolution optical fluctuation imaging (SOFI) now captures multiple focal planes simultaneously. This multiplane 3D SOFI approach enhances depth sampling and reduces imaging time for super-resolution microscopy.

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Area of Science:

  • Optical Microscopy
  • Super-resolution Imaging
  • Biophysics

Background:

  • Super-resolution optical fluctuation imaging (SOFI) overcomes the diffraction limit in 3D using higher-order cumulants of blinking fluorophore images.
  • Previous 3D SOFI required sequential imaging of multiple depth positions, increasing acquisition time and photobleaching.

Purpose of the Study:

  • To introduce a multiplexed imaging scheme for simultaneous acquisition of multiple focal planes in 3D SOFI.
  • To enhance depth sampling and reduce acquisition time and photobleaching in super-resolution microscopy.

Main Methods:

  • Developed a multiplexed imaging scheme for simultaneous multiplane acquisition.
  • Applied 3D cross-cumulants to analyze data from multiple focal planes.
  • Demonstrated the technique on fluorescently labeled cellular structures (mitochondria and vimentin).

Main Results:

  • Achieved increased depth sampling using simultaneous multiplane acquisition.
  • Significantly reduced acquisition time and photobleaching compared to sequential methods.
  • Successfully imaged cellular structures in a volume up to 65 × 65 × 3.5 μm(3) without depth scanning.

Conclusions:

  • Multiplane 3D SOFI enables faster, more efficient super-resolution imaging.
  • The technique allows for high-resolution 3D imaging of cellular structures with reduced photodamage.
  • This advancement is valuable for studying dynamic biological processes in 3D.