Motion-artifact-robust, polarization-resolved second-harmonic-generation microscopy based on rapid polarization switching with electro-optic Pockells cell and its application to in vivo visualization of collagen fiber orientation in human facial skin.
Polarization-resolved second-harmonic-generation (PR-SHG) microscopy is a powerful tool for investigating collagen fiber orientation quantitatively with low invasiveness. However, the waiting time for the mechanical polarization rotation makes it too sensitive to motion artifacts and hence has hampered its use in various applications in vivo. In the work described in this article, we constructed a motion-artifact-robust, PR-SHG microscope based on rapid polarization switching at every pixel with an electro-optic Pockells cell (PC) in synchronization with step-wise raster scanning of the focus spot and alternate data acquisition of a vertical-polarization-resolved SHG signal and a horizontal-polarization-resolved one. The constructed PC-based PR-SHG microscope enabled us to visualize orientation mapping of dermal collagen fiber in human facial skin in vivo without the influence of motion artifacts. Furthermore, it implied the location and/or age dependence of the collagen fiber orientation in human facial skin. The robustness to motion artifacts in the collagen orientation measurement will expand the application scope of SHG microscopy in dermatology and collagen-related fields.
Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan.
Yuji
Y
Tanaka
Graduate School of Advanced Technology and Science, The University of Tokushima, 2-1 Minami-Josanjima, Tokushima 770-8506, Japan.
Eiji
E
Hase
Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan ; Department of Anatomy and Cell Biology, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan.
Shuichiro
S
Fukushima
Shiseido Research Center, 2-2-1 Hayabuchi, Tsuzuki-Ku, Yokohama 224-8558, Japan.
Yuki
Y
Ogura
Shiseido Research Center, 2-2-1 Hayabuchi, Tsuzuki-Ku, Yokohama 224-8558, Japan.
Toyonobu
T
Yamashita
Shiseido Research Center, 2-2-1 Hayabuchi, Tsuzuki-Ku, Yokohama 224-8558, Japan.
Tetsuji
T
Hirao
Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan.
Tsutomu
T
Araki
Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan ; Department of Anatomy and Cell Biology, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan ; Institute of Technology and Science, The University of Tokushima, 2-1 Minami-Josanjima, Tokushima 770-8506, Japan.
Takeshi
T
Yasui
(170.1870) Dermatology; (170.3880) Medical and biological imaging; (180.4315) Nonlinear microscopy; (190.4160) Multiharmonic generation