Multimodal and multiplex spectral imaging of rat cornea ex vivo using a white-light laser source.
We applied our multimodal nonlinear spectral imaging microscope to the measurement of rat cornea. We successfully obtained multiple nonlinear signals of coherent anti-Stokes Raman scattering (CARS), third-order sum frequency generation (TSFG), and second harmonic generation (SHG). Depending on the nonlinear optical processes, the cornea tissue was visualized with different image contrast mechanism simultaneously. Due to white-light laser excitation, multiplex CARS and TSFG spectra were obtained. Combined multimodal and spectral analysis clearly elucidated the layered structure of rat cornea with molecular structural information. This study indicates that our multimodal nonlinear spectral microscope is a promising bioimaging method for tissue study. Multimodal nonlinear spectral images of rat cornea at corneal epithelium and corneal stroma in the in-plane (XY) direction. With use of the combinational analysis of different nonlinear optical processes, detailed molecular structural information is available without staining or labelling.
Department of Chemistry, School of Science, The University of Tokyo, 7-3-1, Hongo, Bunkyo, Tokyo, 113-0033, Japan.
Hiroki
H
Segawa
Graduate School of Comprehensive Human Science, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, Japan.
Yuichi
Y
Kaji
Xlim Research Institute, CNRS-University of Limoges, 123 Avenue Albert Thomas, 87060, Limoges cedex, France.
Philippe
P
Leproux
Xlim Research Institute, CNRS-University of Limoges, 123 Avenue Albert Thomas, 87060, Limoges cedex, France.
Vincent
V
Couderc
Department of Chemistry, School of Science, The University of Tokyo, 7-3-1, Hongo, Bunkyo, Tokyo, 113-0033, Japan.
Takeaki
T
Ozawa
Graduate School of Comprehensive Human Science, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, Japan.
Tetsuro
T
Oshika
Institute of Applied Physics, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8573, Japan. hkano@bk.tsukuba.ac.jp.
Hideaki
H
Kano
D000818:Animals; D003315:Cornea / Q000737:chemistry / Q000166:cytology*; D007834:Lasers; D008027:Light; D008297:Male; D064847:Multimodal Imaging / Q000295:instrumentation*; D017785:Photons; D051381:Rats; D013057:Spectrum Analysis; D014732:Vibration
cornea; molecular imaging; multimodal imaging; spectrum analysis Raman/instrumentation