慶應義塾研究者情報データベース

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• マルチモーダル非線形光学顕微鏡

  

  マウス脳切片のCARSイメージ

  

  生命現象は、その機能発現に様々な生体分子が協働する複雑な物理化学現象です。生細胞内では、このようなダイナミックな現象が“ごく当たり前”に起こっており、その究極的な理解には、機能する分子をそのまま可視化することが必須です。ラマン分光法は、生細胞内にある分子分布や分子構造、動態を、非染色・非標識・非破壊・低侵襲でその場観察可能な、非常に強力な手法の一つです。私たちは、微弱なラマン散乱光を増幅する非線形ラマン散乱を用い、分子集合体から生細胞・生体組織まで、様々な系を「化学の眼」で可視化し、未知の生命現象の発見とその本質の解明に挑んでいます。

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• 博士（理学）, 東京大学, 課程, 2001年03月

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• Coherent Anti-Stokes Raman Scattering (CARS) Microscopy and its Applications in Life Sciences

  Miyazaki S., Murakami Y., Honjoh S., Hayashi Y., Kano H., Raman Spectroscopy in Human Health and Biomedicine, 2023年01月

  　概要を見る

  The following sections are included: Introduction Basic Principles Four Analogs in Coherent Raman Scattering Conditions for CARS Signal Acquisition and CARS Signal Features Early Development of CARS Microscopy Practical Implementation of CARS Microscopy Overview Laser Sources for CARS Microscopy Hyperspectral Imaging Using CARS Microscopy Wavelength Scanning Spectral Focusing Fourier Transformation Multiplex Method Retrieval of a Raman Spectrum from a CARS Spectrum Applications of CARS Microscopy in Life Sciences Monochromatic (Single-Color) CARS Imaging Hyperspectral (Multicolor) CARS Imaging Living Cells Nematodes Mouse Brain Tissues CARS and Pathology In Vivo CARS Imaging CARS and Environmental Science Summary References

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• β-Sheet alignment in intact feathers revealed by polarization-resolved second harmonic generation (SHG) and coherent Raman imaging

  Hu Z., Oketani R., Hiramatsu K., Leproux P., Suzuki R., Kano H.

  Chemistry Letters 54 （ 11 ）  2025年11月

  ISSN  03667022

  　概要を見る

  We visualized β-sheet alignment in intact feathers using polarization-resolved second harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) imaging. SHG signal intensity was strongly dependent on the polarization of the incident laser, while the amide I and III bands in the CARS spectra exhibited moderate polarization-dependent behavior. These results demonstrate that SHG and CARS enable complementary, label-free assessment of structural anisotropy in β-sheet–rich protein fibers, with SHG being especially sensitive to the orientation of the peptide backbone.

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• Chromatic aberration compensation in multiplex coherent Raman microscopy using a Bessel beam

  Oketani R., Tanaka K., Murakami Y., Kano H.

  Optics Letters 50 （ 19 ） 6221 - 6224 2025年10月

  　概要を見る

  Multiplex coherent anti-Stokes Raman scattering (CARS) microscopy enables label-free spectroscopic imaging over an ultra-broadband range using a supercontinuum light source. However, under high numerical aperture conditions, chromatic aberration causes axial splitting of the illumination spot, leading to missing or significantly weakened peaks. Here, we used Bessel beam illumination to compensate for chromatic aberration without modifying objective lenses. Our numerical and experimental results demonstrate that combining the nonlinear nature of the CARS process with the axial elongation of the Bessel beam spot simultaneously achieves broadband spectroscopy and high spatial resolution imaging. This approach is expected to further expand the applicability of vibrational spectroscopic imaging technique.

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• Near-Infrared Coumarin-Hemicyanine Hybrid Dyes Bearing an Intramolecular Nucleophile for Activatable Fluorescence and Raman Imaging

  Fujioka H., Sakamoto R., Hiramatsu K., Murakami Y., Masaki M., Kawatani M., Matsumoto S., Kojima R., Urano Y., Kano H., Kamiya M.

  Analytical Chemistry 97 （ 32 ） 17589 - 17597 2025年08月

  ISSN  00032700

  　概要を見る

  Coumarin-hemicyanine (CHC) hybrid fluorophores bearing an intramolecular nucleophile have been reported as ratiometric fluorescent probes based on the equilibrium between the π-conjugated open and cyclic closed forms. Here, we report red-shifted CHC derivatives that can serve as scaffold dyes of activated fluorescent probes and activated Raman probes. We prepared a series of cyano-substituted CHC derivatives and found that substitution at C-4 of the coumarin moiety is effective in extending the absorption wavelength. The near-infrared (NIR) emission of these 4CN-CHC derivatives exhibits high tissue penetration. Dual-color imaging with 4CN-CHC enabled ratiometric monitoring of the lysosomal pH changes. Furthermore, the coherent anti-Stokes Raman scattering (CARS) signals of 4CN-CHC derivatives could be regulated by modifying the open/closed equilibrium, and the different patterns of CARS spectra enabled us to perform multicomponent imaging. Thus, these 4CN-CHC derivatives expand the repertoire of functional probes for NIR and multiplexed vibrational imaging.

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• Apoptosis induction by ceramide derivatives and its potential mechanisms through domain formation

  Tsujimura K., Yakabe M., Kano H., Matsumori N.

  Bioorganic and Medicinal Chemistry 126 2025年08月

  ISSN  09680896

  　概要を見る

  Ceramides, which are recognized as pivotal signaling agents in cell differentiation and proliferation, elicit apoptosis in response to anticancer drugs and stress. Although the 1-hydroxy group of ceramide is considered to play an important role in inducing apoptosis, the detailed mechanism underlying ceramide-induced apoptosis remains elusive. We previously synthesized ceramide derivatives by transforming the 1-hydroxy group into amino and carboxy groups, and assessed their ability to form domains in artificial membranes. In this study, we evaluated the apoptotic activities of these derivatives against living cells. Surprisingly, despite the lack of the 1-hydroxy group, most of the derivatives exhibited apoptotic activity, with some being more active than ceramide. Confocal microscopy using fluorescent-ceramide and coherent anti-Stokes Raman scattering microscopy utilizing deuterated ceramide suggested the formation of large domains on living cell membranes. These findings suggest a potential relationship between domain formation and apoptotic activity. Liquid chromatography-mass spectrometry showed that the differences in apoptotic activity among the derivatives were unlikely attributed to variations in cellular uptake. Consequently, we propose that these ceramide derivatives accumulate and form distinct domains within the cellular membranes, inducing apoptosis. Intracellular ceramides synthesized in response to external stimuli may trigger apoptosis through the same mechanism.

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• Label-Free Visualization of Ciliary Rootlets in Mouse Brain

  Murakami Y., Nuriya M., Hu Z., Tomioka M., Oketani R., Hiramatsu K., Leproux P., Inoko A., Honjoh S., Kano H.

  Analytical Chemistry 97 （ 27 ） 14160 - 14167 2025年07月

  ISSN  00032700

  　概要を見る

  Neuronal primary cilia play important roles in brain development, sensory perception, and neurogenesis. Rootletin, a fibrous protein composed of coiled-coil motifs, is a key structural component of ciliary rootlets and is essential for understanding ciliary functions. However, the precise mechanisms by which Rootletin influences ciliary dynamics and impacts neuronal function remain largely unknown, primarily due to the challenges associated with visualizing these structures. In this study, we present a label-free, rapid, and highly sensitive method for visualizing Rootletin molecules in brain tissue. This platform integrates a second harmonic generation (SHG) microscope with background reduction by a one-step chemical pretreatment. Additionally, we employed coherent anti-Stokes Raman scattering imaging to simultaneously determine the cellular regions and intracellular locations of SHG signals. Applying this multimodal multiphoton imaging approach to the mouse hippocampus revealed that neuronal ciliary rootlets exhibited highly organized and specific intracellular distributions. Moreover, measurements of cultured neurons revealed that ciliary rootlets were detected even in immature neurons. These findings highlight the utility of our label-free imaging platform in developmental and neuroscience research, providing a powerful tool for characterizing ciliary dynamics and neuronal function.

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• Tribute to Hiro-o Hamaguchi: Expanding the Boundaries of Raman Spectroscopy

  Kano H., Bonn M., Zanni M., Tahara T.

  Journal of Physical Chemistry B （Journal of Physical Chemistry B）  128 （ 4 ） 883 - 885 2024年02月

  ISSN  15206106

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• 心アミロイドーシスの早期診断・早期治療に向けた分光特性と細胞病態の相関理解

  2024年04月

  -

  2027年03月

  加納 英明, 基盤研究(B), 補助金,  研究代表者

• 第二高調波による細胞内らせん構造のラベルフリー新機能探索

  2023年06月

  -

  2025年03月

  加納 英明, 挑戦的研究（萌芽）, 補助金,  研究代表者

担当授業科目 【 表示 ／ 非表示 】

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• 生命システム情報特別講義Ａ

  2025年度

• 生命情報特別講義第１

  2025年度

• 生命情報輪講

  2025年度

• 生命現象の物理学

  2025年度

• 生命系の物理化学第２

  2025年度

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