Adaptive Optical Two-Photon Microscopy for Surface-Profiled Living Biological Specimens
Open accessAbstract
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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Cited by
Sorted by citations
High-Efficiency Polarization-Independent LCoS Utilizing a Silicon-Based Metasurface
Deng, Y., Liu, B., & Yan, J. (2025) · Photonics · doi:10.3390/photonics12060552 · Cited by 1 · see citing works
“…image projections), phase-only modulating LCoS devices [2,3] have a wider range of applications, including correction of chromatic aberration in imaging systems [4], phase control in biological optical microscopy [5], wavefront correction in adaptive optics systems [6,7], and spectral shaping [8].”
- Authors
- Yamaguchi, K., Otomo, K., Kozawa, Y., Tsutsumi, M., Inose, T., Hirai, K., Sato, S., Nemoto, T., & Uji-i, H.
- Affiliations
- National Institutes of Natural Sciences (JP)
- Tohoku University (JP)
- Hokkaido University (JP)
- Hokkaido University , , , ,
- Corresponding author
- Nemoto, Tomomi
- ORCID
- Venue
- ACS Omega
- Publisher
- American Chemical Society (ACS)
- Published
- 2020-11-30
- Volume
- 6
- Issue
- 1
- Pages
- 438-447
- ISSN
- 2470-1343 (ISSN-L)
- PMC
- PMC7807736
- OpenAlex
- W3108360418
- Cited by
- 19
- References
- 40
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 3 · 2023: 3 · 2022: 4 · 2021: 4
- FWCI
- 1.31
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- ACS Omega (publishedVersion, other-oa)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Lirias (KU Leuven) (submittedVersion, cc-by-nc-nd)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Hokkaido University Collection of Scholarly and Academic Papers (Hokkaido University) (submittedVersion)
- Institutional Repositories DataBase (IRDB) (submittedVersion)
- Funders
- Japan Agency for Medical Research and Development (NA)
- Japan Society for the Promotion of Science (JP15H05953, JP16K15103, JP17H03003, JP18K06591, JP19K15406, JP20H00523, JP20K05413, JP20K21165)
- Ministry of Education, Culture, Sports, Science and Technology (NA)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Digital Holography and Microscopy
- Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Adaptive optics and wavefront sensing
- Keywords
- adaptive optics, two-photon microscopy, spatial light modulator, 3D ray tracing, point spread function, fluorescence imaging, curved surfaces, tumor spheroids, dendritic spines, cortical layer V, mouse brain
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