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In situ Imaging of the Mouse Thymus Using 2-Photon Microscopy
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Fast axial scanning for 2-photon microscopy using liquid lens technology.

Kayvan Forouhesh Tehrani1, Min Kyoung Sun1, Lohitash Karumbaiah1

  • 1Regenerative Bioscience Center, Rhodes Center for ADS, University of Georgia, Athens, GA 30602, USA.

Proceedings of Spie--The International Society for Optical Engineering
|May 1, 2018
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Summary

This study introduces liquid lenses for rapid Z-scanning in microscopy, offering a vibration-resistant alternative to mechanical scanning for imaging dynamic biological samples.

Keywords:
Active opticsLiquid lensMulti-photon microscopyRemote focusing

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Area of Science:

  • Optical microscopy
  • Biomedical imaging
  • Materials science

Background:

  • 3D volume imaging in microscopy relies on Z-coordinate focus movement.
  • Mechanical Z scanning, while precise, is slow and sensitive to vibrations, limiting live-cell imaging.
  • Alternative methods use conjugate optics to alter the focal plane without physical movement.

Purpose of the Study:

  • To present and characterize the use of a liquid lens for fast axial scanning in microscopy.
  • To offer a more robust and faster Z-scanning solution compared to traditional mechanical methods.
  • To evaluate the liquid lens's performance in terms of Z displacement, spectral characteristics, and pulse duration.

Main Methods:

  • Utilized a liquid lens based on electrowetting to control the curvature of a liquid-liquid interface.
  • Implemented fast axial scanning by adjusting the liquid lens's focal plane without physical sample or objective movement.
  • Characterized the liquid lens's effects on Z displacement, beam spectral properties, and pulse duration.

Main Results:

  • Demonstrated fast and precise Z-scanning capabilities using the liquid lens.
  • Showcased the liquid lens's advantages over mechanical scanning, including reduced sensitivity to vibrations and stress.
  • Quantified the impact of the liquid lens on beam characteristics and pulse duration.

Conclusions:

  • Liquid lenses provide a viable and advantageous alternative for fast axial scanning in microscopy.
  • This technology enables improved imaging of dynamic, time-varying samples like live cells.
  • Liquid lenses offer a promising approach for advanced microscopy techniques requiring rapid focal plane adjustments.