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Related Experiment Video

Updated: Nov 27, 2025

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Volumetric Lissajous confocal microscopy with tunable spatiotemporal resolution.

Takahiro Deguchi1, Paolo Bianchini1,2, Gemma Palazzolo3

  • 1Nanoscopy & NIC@IIT, Center for Human Technologies, Istituto Italiano di Tecnologia, via E. Melen 83B, 16152 Genoa, Italy.

Biomedical Optics Express
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Summary

We developed a novel volumetric Lissajous confocal microscopy system for fast 3D imaging of dynamic biological systems. This tunable system achieves high-speed volumetric imaging, enabling detailed observation of cellular processes.

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Area of Science:

  • Optical Microscopy
  • Biophysics
  • Cell Biology

Background:

  • Dynamic biological systems require advanced 3D optical microscopy capable of adapting to rapid temporal and spatial changes.
  • Conventional confocal microscopy techniques often lack tunability in acquisition parameters, leading to suboptimal imaging of fast-moving biological processes.
  • Rigid scanning rates and pixel dwell times in existing methods hinder the capture of dynamic events in biological samples.

Purpose of the Study:

  • To develop a volumetric Lissajous confocal microscopy system with enhanced 3D scanning speed and tunable sampling rates.
  • To overcome the limitations of fixed acquisition parameters in traditional microscopy for imaging dynamic biological systems.
  • To enable high-speed, high-resolution 3D imaging of biological processes at the cellular level.

Main Methods:

  • Integration of an acoustic liquid lens for continuous axial focus translation with a resonant scanning mirror.
  • Implementation of a dynamic Lissajous trajectory for the excitation beam, enabling sub-millisecond 3D data acquisition.
  • Utilizing sub-Nyquist sampling rates combined with temporal accumulation and advanced interpolation algorithms for selectable volumetric imaging rates.

Main Results:

  • Demonstrated multicolor and calcium imaging in volumes of tens of cubic microns.
  • Achieved 3D acquisition speeds of 30 Hz and frame rates up to 5 kHz.
  • The developed system offers selectable spatiotemporal resolution through post-processing for specific biological events.

Conclusions:

  • Volumetric Lissajous confocal microscopy provides unprecedented 3D scanning speed and tunability for dynamic biological imaging.
  • The system's flexibility in acquisition and post-processing allows for optimized imaging of fast biological events.
  • This technology advances the capability to study complex cellular dynamics in three dimensions with high temporal resolution.