Special Section on Endomicroscopy Technologies and Biomedical Applications

Increased illumination uniformity and reduced photodamage offered by the Lissajous scanning in fiber-optic two-photon endomicroscopy

[+] Author Affiliations
Wenxuan Liang, Kartikeya Murari, Yuying Zhang, Yongping Chen, Xingde Li

Johns Hopkins University, Department of Biomedical Engineering, Baltimore, Maryland 21205

Ming-Jun Li

Science and Technology Division, Corning Incorporated, SP-AR-02-2, Corning, New York 14831

J. Biomed. Opt. 17(2), 021108 (Feb 27, 2012). doi:10.1117/1.JBO.17.2.021108
History: Received September 22, 2011; Revised January 3, 2012; Accepted January 9, 2012
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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.

Figures in this Article
© 2012 Society of Photo-Optical Instrumentation Engineers

Citation

Wenxuan Liang ; Kartikeya Murari ; Yuying Zhang ; Yongping Chen ; Ming-Jun Li, et al.
"Increased illumination uniformity and reduced photodamage offered by the Lissajous scanning in fiber-optic two-photon endomicroscopy", J. Biomed. Opt. 17(2), 021108 (Feb 27, 2012). ; http://dx.doi.org/10.1117/1.JBO.17.2.021108


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