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Proceedings Paper

Transmissive liquid-crystal device correcting primary coma aberration and astigmatism in laser scanning microscopy
Author(s): Ayano Tanabe; Terumasa Hibi; Sari Ipponjima; Kenji Matsumoto; Masafumi Yokoyama; Makoto Kurihara; Nobuyuki Hashimoto; Tomomi Nemoto
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Paper Abstract

Laser scanning microscopy allows 3D cross-sectional imaging inside biospecimens. However, certain aberrations produced can degrade the quality of the resulting images. We previously reported a transmissive liquid-crystal device that could compensate for the predominant spherical aberrations during the observations, particularly in deep regions of the samples. The device, inserted between the objective lens and the microscope revolver, improved the image quality of fixed-mouse-brain slices that were observed using two-photon excitation laser scanning microscopy, which was originally degraded by spherical aberration. In this study, we developed a transmissive device that corrects primary coma aberration and astigmatism, motivated by the fact that these asymmetric aberrations can also often considerably deteriorate image quality, even near the sample surface. The device's performance was evaluated by observing fluorescent beads using single-photon excitation laser scanning microscopy. The fluorescence intensity in the image of the bead under a cover slip tilted in the y-direction was increased by 1.5 times after correction by the device. Furthermore, the y- and z-widths of the imaged bead were reduced to 66% and 65%, respectively. On the other hand, for the imaged bead sucked into a glass capillary in the longitudinal x-direction, correction with the device increased the fluorescence intensity by 2.2 times compared to that of the aberrated image. In addition, the x-, y-, and z-widths of the bead image were reduced to 75%, 53%, and 40%, respectively. Our device successfully corrected several asymmetric aberrations to improve the fluorescent signal and spatial resolution, and might be useful for observing various biospecimens.

Paper Details

Date Published: 15 March 2016
PDF: 7 pages
Proc. SPIE 9717, Adaptive Optics and Wavefront Control for Biological Systems II, 97171N (15 March 2016); doi: 10.1117/12.2209459
Show Author Affiliations
Ayano Tanabe, Hokkaido Univ. (Japan)
Citizen Holdings Co., Ltd. (Japan)
Terumasa Hibi, Hokkaido Univ. (Japan)
Sari Ipponjima, Hokkaido Univ. (Japan)
Kenji Matsumoto, Citizen Holdings Co., Ltd. (Japan)
Masafumi Yokoyama, Citizen Holdings Co., Ltd. (Japan)
Makoto Kurihara, Citizen Holdings Co., Ltd. (Japan)
Nobuyuki Hashimoto, Citizen Holdings Co., Ltd. (Japan)
Tomomi Nemoto, Hokkaido Univ. (Japan)


Published in SPIE Proceedings Vol. 9717:
Adaptive Optics and Wavefront Control for Biological Systems II
Thomas G. Bifano; Joel Kubby; Sylvain Gigan, Editor(s)

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