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Updated: Jan 24, 2026

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Lightsheet localization microscopy enables fast, large-scale, and three-dimensional super-resolution imaging.

Chieh-Han Lu1, Wei-Chun Tang1, Yen-Ting Liu1

  • 11Research Center for Applied Sciences, Academia Sinica, Taipei, 11529 Taiwan.

Communications Biology
|May 18, 2019
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Researchers developed a fast, large-scale 3D super-resolution microscope using Bessel lightsheet imaging. This breakthrough enables high-resolution visualization of multiple whole cells, advancing biological structure discovery.

Keywords:
Nuclear envelopeSuper-resolution microscopy

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Area of Science:

  • Biophysics
  • Microscopy
  • Cell Biology

Background:

  • Current super-resolution microscopy struggles with imaging multiple whole cells due to weak signals and slow acquisition.
  • Limitations hinder the detailed study of complex cellular structures in their native environments.

Purpose of the Study:

  • To develop a fast, large-scale, 3D super-resolution fluorescence microscope.
  • To overcome limitations in imaging multiple whole cells.
  • To enable new insights into complex biological structures and live cell imaging.

Main Methods:

  • Utilized a single-wavelength Bessel lightsheet illumination strategy.
  • Selective illumination of spontaneous blinking fluorophores tagged to proteins.
  • Systematic optimization of labeling density, excitation power, and exposure time.

Main Results:

  • Achieved a maximum imaging speed of 2.7 × 10^4 µm³ s⁻¹.
  • Reconstructed 3D super-resolution images with a lateral resolution of approximately 75 nm.
  • Demonstrated imaging of 250 volumes on immobilized samples within 15 minutes.

Conclusions:

  • The developed Bessel lightsheet microscope offers unprecedented speed and scale for 3D super-resolution imaging.
  • This system facilitates the discovery of complex biological structures.
  • Opens possibilities for live 3D localization imaging within multiple whole cells.