TY - JOUR
T1 - Increased illumination uniformity and reduced photodamage offered by the Lissajous scanning in fiber-optic two-photon endomicroscopy
AU - Liang, Wenxuan
AU - Murari, Kartikeya
AU - Zhang, Yuying
AU - Chen, Yongping
AU - Li, Ming Jun
AU - Li, Xingde
N1 - Funding Information:
The authors gratefully thank Jiefeng Xi for his useful discussions on scanner calibration. This research was supported in part by the National Institutes of Health under grants R01 CA153023 and R01 EB007636 and the Individual Biomedical Research Award (to XDL) from The Hartwell Foundation.
PY - 2012/2
Y1 - 2012/2
N2 - 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.
AB - 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.
KW - Edomicroscopy
KW - Sanning probe.
KW - To-photon fluorescence
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U2 - 10.1117/1.JBO.17.2.021108
DO - 10.1117/1.JBO.17.2.021108
M3 - Article
C2 - 22463026
AN - SCOPUS:84864529547
VL - 17
JO - Journal of Biomedical Optics
JF - Journal of Biomedical Optics
SN - 1083-3668
IS - 2
M1 - 021108
ER -