TY - JOUR
T1 - Adaptive Optical Two-Photon Microscopy for Surface-Profiled Living Biological Specimens
AU - Yamaguchi, Kazushi
AU - Otomo, Kohei
AU - Kozawa, Yuichi
AU - Tsutsumi, Motosuke
AU - Inose, Tomoko
AU - Hirai, Kenji
AU - Sato, Shunichi
AU - Nemoto, Tomomi
AU - Uji-I, Hiroshi
N1 - Funding Information:
We thank Dr. Ryosuke Enoki and Dr. Hirokazu Ishii of the Division of Biophotonics at the National Institute for Physiological Sciences, National Institute of Natural Sciences for their helpful advice regarding the experimental set up. We are also grateful to Dr. Kentaro Kobayashi and Dr. Yasutaka Matsuo of the Nikon Imaging Center at Hokkaido University for providing technical support. This collaborative work was partially supported by the JSPS “Core-to-Core Program, A. Advanced Research Networks”, the Research Program of “Five-star Alliance” in “NJRC Mater. & Dev.” (MEXT), “Network Joint Research Center for Materials and Devices”, and “the Photo-excitonix in Hokkaido University”. We would like to thank Enago ( www.enago.jp ) for the English language review.
Publisher Copyright:
© 2021 ACS. All rights reserved.
PY - 2021/1/12
Y1 - 2021/1/12
N2 - We developed adaptive optical (AO) two-photon excitation microscopy by introducing a spatial light modulator (SLM) in a commercially available microscopy system. For correcting optical aberrations caused by refractive index (RI) interfaces at a specimen's surface, spatial phase distributions of the incident excitation laser light were calculated using 3D coordination of the RI interface with a 3D ray-tracing method. Based on the calculation, we applied a 2D phase-shift distribution to a SLM and achieved the proper point spread function. AO two-photon microscopy improved the fluorescence image contrast in optical phantom mimicking biological specimens. Furthermore, it enhanced the fluorescence intensity from tubulin-labeling dyes in living multicellular tumor spheroids and allowed successful visualization of dendritic spines in the cortical layer V of living mouse brains in the secondary motor region with a curved surface. The AO approach is useful for observing dynamic physiological activities in deep regions of various living biological specimens with curved surfaces.
AB - We developed adaptive optical (AO) two-photon excitation microscopy by introducing a spatial light modulator (SLM) in a commercially available microscopy system. For correcting optical aberrations caused by refractive index (RI) interfaces at a specimen's surface, spatial phase distributions of the incident excitation laser light were calculated using 3D coordination of the RI interface with a 3D ray-tracing method. Based on the calculation, we applied a 2D phase-shift distribution to a SLM and achieved the proper point spread function. AO two-photon microscopy improved the fluorescence image contrast in optical phantom mimicking biological specimens. Furthermore, it enhanced the fluorescence intensity from tubulin-labeling dyes in living multicellular tumor spheroids and allowed successful visualization of dendritic spines in the cortical layer V of living mouse brains in the secondary motor region with a curved surface. The AO approach is useful for observing dynamic physiological activities in deep regions of various living biological specimens with curved surfaces.
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U2 - 10.1021/acsomega.0c04888
DO - 10.1021/acsomega.0c04888
M3 - Article
AN - SCOPUS:85097732690
SN - 2470-1343
VL - 6
SP - 438
EP - 447
JO - ACS Omega
JF - ACS Omega
IS - 1
ER -