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27 February 2012 Increased illumination uniformity and reduced photodamage offered by the Lissajous scanning in fiber-optic two-photon endomicroscopy
Wenxuan Liang, Kartikeya Murari, Yuying Y. Zhang, Yongping Chen, Xingde D. Li, Ming-Jun Li
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Journal of Biomedical Optics, Vol. 17, Issue 2, 021108 (February 2012). https://doi.org/10.1117/1.JBO.17.2.021108
Abstract
We compare the illumination uniformity and the associated effects of the spiral and Lissajous scanning patterns that are commonly used in an endomicroscope. Theoretical analyses and numerical simulations were first performed to quantitatively investigate the area illumination density in the spiral scanning pattern. The results revealed the potential problem of manifest photodamage due to the very high illumination density in the center of the spiral scan. Similar analyses of the Lissajous scanning pattern, which can be conveniently implemented on the same endomicroscope with no hardware modifications, showed a more uniform illumination density with about an 80-fold reduction in the peak illumination density. To underscore the benefit offered by the improved illumination uniformity, we conducted in vitro two-photon fluorescence imaging of cultured cells stained with a LIVE/DEAD viability assay using our home-built, fiber-optic, two-channel endomicroscopy system. Both the spiral and the Lissajous scans were implemented. Our experimental results showed that cells near the spiral scan center experienced obvious photodamage, whereas cells remained alive over the entire region under the Lissajous beam scanning, confirming the predicted advantage offered by the Lissajous scan over this spiral scan in an endomicroscopy setting.
© 2012 Society of Photo-Optical Instrumentation Engineers (SPIE) 0091-3286/2012/$25.00 © 2012 SPIE
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Wenxuan Liang, Kartikeya Murari, Yuying Y. Zhang, Yongping Chen, Xingde D. Li, and Ming-Jun Li "Increased illumination uniformity and reduced photodamage offered by the Lissajous scanning in fiber-optic two-photon endomicroscopy," Journal of Biomedical Optics 17(2), 021108 (27 February 2012). https://doi.org/10.1117/1.JBO.17.2.021108
Published: 27 February 2012
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KEYWORDS
Luminescence
Fiber optics
Endomicroscopy
Ferroelectric materials
Electrodes
Numerical simulations
Data acquisition
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