nanophotonics, members, 日本語
Katsumasa FUJITA

Ph. D.
Professor, Department of Applied Physics, Osaka University
2-1 Yamadaoka, Suita, Osaka 565-0871, Japan
office: P2-300, Suita Campus, Osaka University
phone: +81-6-6879-7847
email: fujita@ap.eng.osaka-u.ac.jp

BRI Workshop 2024

 We will organize Biomedical Raman Imaging Workshop 2024on 25 - 27 November 2024 in Osaka. The workshop includes lectures about the principle and applications of Raman imaging for biomedical researches and demonstration of cell imaging using facilities in our lab. The event will be jointly organized with The 22nd Annual Meeting of the Japan Association of Medical Spectroscopy. The due dates for abstract submission/participation are 20 October / 7 November 2024.

BRI Symposium 2019

 We organized a symposium: Biomedical Raman Imaging 2019 on 24 - 26 Nov, 2019 in Osaka. There were 129 participants from many different countries!

A report in Spectroscopy, 35 (4), 43–46 (2020).


Research Topics

HeLa cell stained with AttoR6G phalloidin
 Super-resolution fluorescence microscopy
We are developing optical microscopy techniques which can resolve fine structures of samples with the spatial resolution beyond the diffraction limit of light. By using saturated excitation or photoswiching of fluorescent molecules, we induce nonlinear fluorescence respoinse to realize super-resolution imaging. For example, saturated exction (SAX) miroscope can be realized by simply modulating laser intensity and demodulating fluorescence signal in a typical confocal microscope.

The figure shows the fluorescence images of a HeLa cell stained with AttoR6G phalloidin (xz cross-section). SAX microscopy can image fine structures that can not be resolved by conventional confocal microscopy.

References:
Temma et al., Nat. Methods (2024).
Temma et al., Opt. Express (2022).
Nawa et al., APL Photonics (2018).
Oketani et al., Opt. Lett. (2017)
Yonemaru et al., Phys. Rev. Applied (2015).
Chu et al., Phys. Rev. Lett. (2014).
Yamanaka et al., J. Biomed. Opt. (2013).
Fujita et al., Phys. Rev. Lett. (2007)

753 cm-1 : Cytochrome
1686 cm-1 : Protein
2852 cm-1 : Lipid
Movie by Raman
Cell division: cytokinesis
Cell division: mitosis
 Raman microscopy for imaging of biological samples
We are developing Raman microscopes to image and analyze biomolecules in living cells. Raman scattering spectra show molecular vibrations in a sample, that contain rich information about species, conditions and environments of molecules. We have developed a slit-scanning Raman microscope and applied it to imaging of biomolecule dynamics in a living cell. We are also developing a higher-sensitive Raman detection technique for cellular imaging by using surface-enhanced Raman scattering and Raman tags to image small molecules.

The figure shows a Raman image of living HeLa cells. Molecular vibrations detected by Raman spectra produce the distribution of proteins and lipids in the cells.

References:
Bando et al., Biomed. Opt. Express. (2022).
SPIE Newroom (2016).
Wanatabe et al., Nat. Commun. (2015).
Palonpon et al, Nat. Protoc. (2013).
Okada et al., PNAS (2012).
Ando et al., Nano Lett. (2011).
Hamada et al., J. Biomed. Opt. (2008)


Alkyne (EdU), Cytochrome , Lipid



 Raman tag imaging
Raman tag imaging is a new technique to observe small molecules in living cells and tissues. Small molecules are too "small" to be labeled by fluorescent dyes and could not be observed in the physiological conditions. We have demonstrated the use of Raman tag to visualize small molecules. Raman tag, such as an alkyne, shows a Raman band district from intracellular molecules and allows us to observe small molecules labeled by Raman tags with Raman microscopy. The Raman tag also realizes multiplexed imaging of many different molecules via the extremely narrow emission band of Raman scattering. We are also working on the application of the Raman tag technique for pharmaceutical researches.

References:
Dodo et al, JACS (2022).
Koike et al, ACS Nano (2020).
Ando et al, JACS (2016).
Ando et al, PNAS (2015).
Yamakoshi et al, Bioorg. Med. Chem. Lett. (2015).
Yamakoshi et al, Chem Commun. (2014).
Palonpon et al, Nat. Protoc. (2013).
Yamakoshi et al., JACS (2011)

Original Papers
  • K. Temma, R, Oketani, T. Kubo, K. Bando, S. Maeda, K. Sugiura, T. Matsuda, R. Heintzmann, T. Kaminishi, K. Fukuda, M. Hamasaki, T. Nagai, and K. Fujita, "Selective-plane-activation structured illumination microscopy," Nat. Methods (online).
  • A. J. Hobro, N. Pavillon, K. Koike, T. Sugiyama, T. Umakoshi, P. Verma, K. Fujita, and N. I. Smith, "Imaging vs Nonimaging Raman Spectroscopy for High-Throughput Single-Cell Phenotyping," Anal. Chem., 96, 18, 7047–7055 (2024).
  • K. Tabata, H. Kawagoe, J. N. Taylor, K. Mochizuki, T. Kubo, J.-E. Clement, Y. Kumamoto, Y. Harada, A. Nakamura, K. Fujita, and T. Komatsuzaki, "On-the-fly Raman microscopy guaranteeing the accuracy of discrimination," Proc. Natl. Acad. Sci. USA, 121 (12) e2304866121 (2024).
  • T. Hayashi, N. Ito, K. Tabata, A. Nakamura, K. Fujita, Y. Harada, T. Komatsuzaki, "Gaussian process classification bandits," Pattern Recognition, 149, 110224 (2024).
  • H. Yamakoshi, D. Shibata, K. Bando, S. Kajimoto, A. Kohyama, S. Egoshi, K. Dodo, Y. Iwabuchi, M. Sodeoka, K. Fujita, T. Nakabayashi, "Ratiometric analysis of reversible thia-Michael reactions using nitrile-tagged molecules by Raman microscopy," Chem. Commun. 59, 14563-14566 (2023).
  • H.-X. Liao, K. Bando, M. Li, K. Fujita, "Multifocal Raman spectrophotometer for examining drug-Induced and chemical-Induced cellular changes in 3D cell spheroids," Anal. Chem. 95 (39), 14616–14623 (2023).
  • J. N. Taylor, A. Pélissier, K. Mochizuki, K. Hashimoto, Y. Kumamoto, Y. Harada, K. Fujita, T. Bocklitz, T. Komatsuzaki, "Correction for Extrinsic Background in Raman Hyperspectral Images," Anal. Chem., 95 (33) 12298-12305 (2023).
  • K. Temma, M. Wincott, K. Fujita, and M. J. Booth, "Deflectometry based calibration of a deformable mirror for aberration correction and remote focusing in microscopy," Opt. Express., 31 (17) 28503-28514 (2023).
  • A. H. Bhuiyan, J. Clement, Z. Ferdous, K. Mochizuki, K. Tabata, J. N. Taylor, Y. Kumamoto, Y. Harada, T. Bocklitz, K. Fujita, and T. Komatsuzaki, "Differentiability of cell types enhanced by detrending non-homogeneous pattern in line-illumination Raman microscope," Analyst, 148, 3574-3583 (2023).
  • M. Li, Y. Ueyama-Toba, M. Lindley, G. Kongklad, Y. Nawa, Y. Kumamoto, S. Ishida, Y. Kanda, S. Fujita, H. Mizuguchi, K. Fujita, "Label-free evaluation of maturation and hepatotoxicity of human iPSC-derived hepatocytes using hyperspectral Raman imaging," Anal. Chem., 95 (24), 9252–9262 (2023).
  • T. Li, J. Liu, M. Guo, F-C. Bin, J-Y. Wang, A. Nakayama, W-C. Zhang, F. Jin, X-Z. Dong, K. Fujita, M-L. Zheng, "Synthesis of biocompatible BSA-GMA and two-photon polymerization of 3D hydrogels with free radical type I photoinitiator," Int. J. Bioprinting. 9(5), 752 (2023).
  • K. M. Helal, H. Cahyadi, J. N. Taylor, A. Okajima, K. Tabata, Y. Kumamoto, K. Mochizuki, Y. Itoh, T. Takamatsu, H. Tanaka, K. Fujita, T. Komatsuzaki, Y. Harada, "Raman imaging of rat non-alcoholic fatty liver tissues reveals distinct biomolecular states," FEBS Lett., 597, 1517-1527 (2023).
  • K. Mochizuki, Y. Kumamoto, S. Maeda, M. Tanuma, A. Kasai, M. Takemura, Y. Harada, H. Hashimoto, H. Tanaka, N. I. Smith, and K. Fujita, "High-throughput line-illumination Raman microscopy with multislit detection," Biomed. Opt. Express., 14 (3), 1015-1026 (2023).
  • M. Hayakawa, J. N. Taylor, R. Nakao, K. Mochizuki, Y. Sawai, K. Hashimoto, K. Tabata, Y. Kumamoto, K. Fujita, E. Konishi, S. Hirano, H. Tanaka, and T. Komatsuzaki, "Lipid Droplet Accumulation and Adipophilin Expression in Follicular Thyroid Carcinoma," Biochem. Biophys. Res. Commun., 640, 192-201 (2023).


Review Articles and Others


Invited Talks (International)
  • K. Fujita, "High-sensitivity Raman imaging of cryofixed biological samples," ACS Fall Meeting (Denver, Aug 2024).
  • K. Fujita, "Cryo-Raman Imaging: capturing the moment of biological activities," The XXVIII International conference on Raman spectroscopy (ICORS 2024) (Rome, 2 Aug 2024).
  • K. Fujita, "Raman imaging: Principle, Application, and Issues," CHARISMA Raman School 2024 (Rome, 28 Jul 2024).
  • K. Fujita, "Cryo-optical microscopy for multimodal molecular imaging," The Oxford International Conference on Advanced Optics and Photonics (Oxford, Julu 2024)
  • K. Fujita, Cryo-optical microscopy for super-resolution imaging of the moment"," The 21st Congress of the International Union for Pure and Applied Biophysics (IUPAB2024) (Kyoto, Jun 2024).
  • K. Fujita, "Raman microscopy: a new imaging modality for biology and medicine," OPTICA Student Chapter Lecture (Manipal, April 2024).
  • K. Fujita, "Multimodal cryo-optical microscopy for high-sensitivity molecular imaging," Taiwan Japan Bilateral Symposium on Photonics 2024 (Taipei, 21 Feb 2024)
  • K. Fujita, Kenta Mizushima, Shoko Tamura, Masahito Yamanaka, Menglu Li, Yoshinori Harada, Nicholas I. Smith, Yasuaki Kumamoto, Hideo Tanaka "Raman imaging of cryofixed cells with preservation of molecular states," SPIE Photonics West (San Francisco, 27 Jan 2024).
  • K. Fujita, "Visualization of drug uptake and its cellular response by Raman microscopy," SPIE Photonics West (San Francisco, 27 Jan 2024).
  • K. Fujita, "Unraveling of Raman Spectroscopy for Chemical Biology Research," ACS Monash University Malaysia International Student Chapter Seminar Series (Online, 22 Jan 2024).
  • K. Fujita, "High-sensitiivity Raman imaging of cryofixed biological samples," RamanFest2023 (Paris, 9-10 November 2023).
  • K. Fujita, "Enhancement of intraspheroid imaging through Bessel-beam side-illumination in Raman and structured illumination microscopy," International Forum on Microscopy 2023 (Zhongshan, 10 Septmber 2023).
  • K. Fujita, "High-throughput Raman microscopy for detecting intracellular molecules," The 12th Asia-Pacific Laser Symposium (Hakodate, 5 September 2023).
  • K. Fujita, "Hyperspectral Raman microscopy: toward visualization of intracellular chemistry," Gordon Research Conference, Chemical Imaging (Easton, 31 July 2023).
  • K. Fujita, "Improvement of the detection sensitivity in Raman microscopy," Biomedical Raman Imaging 2023 (Atlanta, 27 June 2023).
  • K. Fujita, "Raman microscopy: a new imaging modality for biology and medicine," OIST-OU Joint Symposium, A recipe for scientific synergy -Series 4- “Advancing biotechnology through multidisciplinary approaches” (Suita, 29 May 2023).
  • K. Fujita, "Side-illumination Raman microscopy using a Bessel beam for observation of cell spheroids," The 5th Global Conference on Biomedical Engineering (Taipei, 16 Dec 2023).

    ..... and more


Books
  • H. Yamakoshi, J. Ando, S. Egoshi, K. Dodo, M. Sodeoka, K. Fujita, "Raman Imaging and Screening of Bioactive Small Molecules," Raman Spectroscopy in Human Health and Biomedicine (H. Sato et al Ed, WSPC, September 2023)
  • H. Yamakoshi, K. Fujita, “Spontaneous Raman and SERS microscopy for Raman tag imaging,” in Stimulate Raman Scattering Microscopy, pp. 275-287 (J.-X. Chen et al Ed, Elsevier 2021).
  • K. Fujita, "Super-Resolution Imaging in Raman Microscopy" in Label-Free Super-Resolution Microscopy, Astratov Ed. (Springer Nature, 2019).
  • K. Mochizuki, N. I. Smith, K. Fujita, "Raman microscopy," Reference Module in Chemistry, Molecular Sciences and Chemical Engineering (Elsevier)
  • J. Ando, K. Dodo, K. Fujita, M. Sodeoka, "Visualizing Bioactive Small Molecules by Alkyne Tagging and Slit-Scanning Raman Microscopy, "Visualizing bioactive small molecules by alkyne tagging and slit-scanning Raman microscopy," Systems Chemical Biology: Methods and Protocols, Methods in Molecular Biology, Vol.1888, pp. 99-114 (Springer Nature 2019).
  • K. Fujita, "Micro-Raman Spectroscopy," in Compendium of Surface and Interface Analysis, pp. 375-379 (Springer Singapore, 2018).
  • K. Fujita, K. Mochizuki, N. I. Smith, "SAX microscopy and its application to imaging of 3D cultured cells," in Super-resolution imaging in biomedicine (A. Diaspro and M. A. M. J. van Zandvoort Eds., CRC Press, October 14, 2016).
  • M. Hashimoto, T. Ichimura, K. Fujita, "CARS Microscopy: Implementation of Nonlinear Vibrational Spectroscopy for Far-Field and Near-Field Imaging," in Raman Imaging, Springer Series in Optical Sciences 168 (A. Zoubir Ed., Springer-Verlag Berlin Heidelberg 2012).
  • Yasuaki Kumamoto, Nicholas Isaac Smith, Katsumasa Fujita, Jun Ando and Satoshi Kawata, "Optical Techniques for Future Pacemaker Technology," in Modern Pacemakers - Present and Future (M. K. Da ed. InTech 2011).
  • N. I. Smith, S. Kawano, M. Yamanaka, and K. Fujita, "Nonlinear Fluorescence Imaging by Saturated Excitation," in Nanoscopy and Multidimensional Optical Fluorescence Microscopy, pp. 2-1 ~ 2-16 (A. Diaspro Ed., Chapman and Hall/CRC press, April 2010).
  • P. Verma, K. Fujita, T. Ichimura, and S. Kawata, "Raman, CARS and near-field Raman-CARS microscopy for cellular and molecular imaging," in "Nano Biophotonics - Science and Technology -," pp. 57-71 (H. Masuhara, S. Kawata, and F. Tokunaga Ed. Elservier B.V., Amsterdam, 2007).
  • K. Fujita, N. Smith, and O. Nakamura, "Nonlinear optical imaging and stimulation of living cells," in Nanophotonics -Intengrating photochemistry, Optics and Nano/Bio Materials Studies - (H. Masuhara and S. Kawata Ed. Elsevier B.V., Amsterdam, 2004).
  • K. Fujita and T. Takamatsu, "Real-time in situ calcium imaging with single and two-photon confocal microscopy," in Confocal and two-photon microscopy: Foundation, Application and Advances (A. Diaspro Ed. John Wiley & Sons, Inc., New York, 2001).

    ..... and more

Award


Miscellaneous
  • "Technology development could bring Raman microscopy to the clinic," Optica New Release, Feburuary 6, 2023
  • “A further leap of biomedical Raman imaging,” Spectroscopy, Vol. 35, Issue 7, p.10 (2020).
  • “Technology Feature, Live-cell imaging: Deeper, faster, wider,” Science, Vol. 359, Issue 6383, pp. 1549 (2018).
  • "Raman Imaging of Live Cells," Spectroscopy, 18 Nov (2014).
  • "Harmonic Microscopy," RIKEN Research 14 Mar (2008).
  • Richard Gaughan, "Laser Technique Monitors Calcium," PHOTONICS SPECTRA, January, p.65 (2002).
  • Richard Gaughan, "Near-infrared laser generates calcium waves," Biophotonics International, March, p.54 (2002).