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Video-rate two-photon microendoscopy using second harmonic resonance fiber scanning.

Kai Zhang1,2, Guillaume Ducourthial1, Taresh Sharan1,2,3

  • 1Department of Physics, Northeastern University, Boston, MA 02115, USA.

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|November 18, 2024
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We developed a compact, hand-held two-photon microendoscope capable of video-rate imaging. This advanced fiber scanning technology minimizes motion artifacts for clear tissue visualization.

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Area of Science:

  • Biomedical Optics
  • Microscopy Engineering
  • Medical Imaging Technology

Background:

  • Traditional microendoscopy often suffers from motion artifacts, limiting real-time in vivo imaging.
  • Existing fiber scanning techniques have limitations in frame rate, hindering dynamic tissue observation.

Purpose of the Study:

  • To develop a high-frame-rate, hand-held two-photon microendoscope for artifact-free tissue imaging.
  • To enhance fiber scanning methods for improved imaging speed without compromising resolution.

Main Methods:

  • A 2.5-mm resonant fiber scanning two-photon microendoscope with a 30-mm rigid probe was designed.
  • Higher-order harmonic oscillation scanning techniques were implemented to achieve video-rate imaging (20 Hz).
  • 3D printed mounts were utilized for cost-effective and stable fabrication of the fiber scanner.

Main Results:

  • The microendoscope achieved a 20 Hz frame rate, significantly faster than previous designs.
  • Maintained field-of-view (~125 µm) and optical resolution (1.2 µm lateral, 10.9 µm axial).
  • Demonstrated high stability (<0.3 µm drift over 84 hours) with cost-effective 3D printed components.

Conclusions:

  • The developed microendoscope enables real-time, hand-held imaging of tissues with reduced motion artifacts.
  • This technology advances multiphoton microendoscopy for in vivo applications.
  • The use of higher-order harmonic scanning and 3D printing offers a practical solution for high-performance microendoscopy.