慶應義塾研究者情報データベース

総合紹介 【 表示 ／ 非表示 】

総合紹介 【 表示 ／ 非表示 】

• 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。

学位 【 表示 ／ 非表示 】

学位 【 表示 ／ 非表示 】

• 博士（理学）, 東京大学, 課程, 2001年03月

著書 【 表示 ／ 非表示 】

著書 【 表示 ／ 非表示 】

• 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

  • Access to Document (DOI)

論文 【 表示 ／ 非表示 】

論文 【 表示 ／ 非表示 】

• Monitoring Enzymatic Reactions through Probing Chemical Bond Changes by the CD<inf>3</inf> Group Using Multiplex Coherent Anti-Stokes Raman Scattering

  Imai R., Kano H., Hattori T.

  Analytical Chemistry 97 （ 15 ） 8322 - 8328 2025年04月

  ISSN  00032700

  　概要を見る

  We propose a method to measure enzymatic reactions using the C-D stretching vibrational spectra of CD3 groups that exist next to the site where a chemical bond changes. In the proposed method, temporal changes in the concentrations of the substrate and product are measured from changes in the C-D stretching spectra of the CD3 groups. Vibrational spectra are obtained using multiplex coherent anti-Stokes Raman scattering (CARS) spectroscopy, which can obtain vibrational spectra much faster than spontaneous Raman spectroscopy. We tested the proposed method by examining the dehydrogenation reaction of isopropyl alcohol-1,1,1,3,3,3-D6 (IPA-D6) catalyzed by alcohol dehydrogenase and found that the change in concentrations of the substrate and product, IPA-D6 and acetone-D6, respectively, was successfully measured from the C-D stretching spectra.

  • Access to Document (DOI)

• CMOS and CCD Detection in Raman Spectroscopy: A Comparison Using Spontaneous and Multiplex Coherent Anti-Stokes Raman Scattering (CARS)

  Klement W. . ., Leproux P., Browne W., Kano H.

  Journal of Raman Spectroscopy 2025年

  ISSN  03770486

  　概要を見る

  Cooled CCD cameras are used widely in spectroscopy, mainly due to their sensitivity, low noise under low-light conditions, and relatively high image and spectral readout rates. Despite their many advantages, CCD cameras have limitations. Particularly under bright-light conditions, such as those encountered with coherent Raman spectroscopies, the finite readout time of CCD chips becomes limiting. Furthermore, when weak signals need to be observed close to intense signals, blooming and smearing limit the signal-to-noise ratios achievable. Scientific complementary mixed oxide (sCMOS)–based sensors are relatively new. Although they still show much higher readout noise than cooled CCDs, their application to spectroscopy is of interest given the higher readout rates and dynamic ranges possible. Here, we evaluate sCMOS sensors for multiplex (50 ps) CARS and spontaneous Raman spectroscopy, making a direct comparison with a state-of-the-art EM-CCD detector. The EM-CCD camera outperforms the sCMOS camera in terms of limits of detection, while the sCMOS camera performs better than the CCD in terms of dynamic range and readout rate. Importantly, an sCMOS camera does not suffer from interference due to blooming and smearing seen with CCD cameras, which enables observation of weak bands (e.g., Raman overtones) close to much more intense signals. Here, we show that, at readout rates > 20–50 Hz, the performances of the two detector types are not substantially different. We anticipate that sCMOS-based cameras will find application for bright spectroscopies, such as multiplex CARS, as well as spontaneous Raman spectroscopy and Raman spectral imaging.

  • Access to Document (DOI)

• ms-Time Resolution Raman Spectroscopy Using sCMOS Cameras

  Carpenter A.P., Goulden J., Varagnat A., Klement N., Browne W.R., Kano H.

  Proceedings of SPIE - The International Society for Optical Engineering 13348 2025年

  ISSN  0277786X

  　概要を見る

  Raman spectroscopy is a powerful tool that has found broad adoption in characterizing biological and material systems due to the molecular information embedded in Raman spectra. Some ways this spectroscopic method has been applied include monitoring chemical reactions, identifying molecular structures, and measuring the distribution of chemical species throughout a sample. Traditionally, CCD cameras have found popular use as a detector in Raman spectrometers because spectra can be detected in a single acquisition by dispersing the spectrum across the CCD’s pixel array. While CCDs have aided in the development of Raman spectroscopy as a sensitive detector, their readout mechanism can also introduce experimental limitations through slow frame rates, signal smearing, and blooming of strong signals. We discuss the integration of a scientific complementary metal oxide semiconductor (sCMOS) camera into a Raman spectrometer and conduct a side-by-side comparison with CCD and EMCCD cameras under a range of experimental conditions. The sCMOS readout structure provides performance advantages in bright conditions and enables the rapid acquisition of Raman spectra with millisecond exposure times. Additional, recent, experiments are reviewed that highlight the capability of sCMOS cameras to accurately capture spatially resolved Raman spectra under conditions where other detection technologies might struggle.

  • Access to Document (DOI)

• Exploring liquid–liquid phase separation in vitro and in vivo using multimodal nonlinear optical imaging

  Murakami Y., Obuchi M., Kamizawa H., Miyazaki S., Kishimura A., Oketani R., Hiramatsu K., Leproux P., Hayashi Y., Shiraki K., Kano H.

  Analytical Sciences 2025年

  ISSN  09106340

  　概要を見る

  Liquid–liquid phase separation leads to the formation of liquid droplets (LqDs) such as P granules in Caenorhabditis elegans (C. elegans). In this study, we demonstrate the label-free visualization of LqDs using multimodal nonlinear optical imaging both in vitro and in vivo. In vitro measurements with polymerized adenine [poly(A)], we found significantly higher poly(A) concentrations in LqDs compared to surrounding solutions, with the limit of detection (LoD) of 32 mg/mL. In vivo measurements, we performed label-free imaging of C. elegans. Despite efforts to detect P granules within P lineage cells in both wild-type C. elegans and green fluorescent protein (GFP)-tagged strains, no clear RNA-specific signals were observed. This indicates that the RNA concentration in P granules is lower than anticipated and falls below our in vitro LoD. These results underscore the challenges of label-free RNA detection in P granules.

  • Access to Document (DOI)

• Coherent Raman microscopy visualizes ongoing cellular senescence through amide I peak shifts originating from β sheets in disordered nucleolar proteins

  Ishibashi S., Inoko A., Oka Y., Leproux P., Kano H.

  Scientific Reports 14 （ 1 ）  2024年12月

  　概要を見る

  Cellular senescence occurs through the accumulation of many kinds of stresses. Senescent cells in tissues also cause various age-related disorders. Therefore, detecting them without labeling is beneficial for medical research and developing diagnostic methods. However, existing biomarkers have limitations of requiring fixation and labeling, or their molecular backgrounds are uncertain. Coherent anti-Stokes Raman scattering (CARS) spectroscopic imaging is a novel option because it can assess and visualize molecular structures based on their molecular fingerprint. Here, we present a new label-free method to visualize cellular senescence using CARS imaging in nucleoli. We found the peak of the nucleolar amide I band shifted to a higher wavenumber in binuclear senescent cells, which reflects changes in the protein secondary structure from predominant α-helices to β-sheets originating from amyloid-like aggregates. Following this, we developed a procedure that can visualize the senescent cells by providing the ratios and subtractions of these two components. We also confirmed that the procedure can visualize nucleolar aggregates due to unfolded/misfolded proteins produced by proteasome inhibition. Finally, we found that this method can help visualize the nucleolar defects in naïve cells even before binucleation. Thus, our method is beneficial to evaluate ongoing cellular senescence through label-free imaging of nucleolar defects.

  • Access to Document (DOI)

全件表示 >>

総説・解説等 【 表示 ／ 非表示 】

総説・解説等 【 表示 ／ 非表示 】

• 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

  • Access to Document (DOI)

競争的研究費の研究課題 【 表示 ／ 非表示 】

競争的研究費の研究課題 【 表示 ／ 非表示 】

• 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 】

担当授業科目 【 表示 ／ 非表示 】

• 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

全件表示 >>