研究者データベース

橋本 守(ハシモト マモル)
情報科学研究院 生命人間情報科学部門 バイオエンジニアリング分野
教授

基本情報

所属

  • 情報科学研究院 生命人間情報科学部門 バイオエンジニアリング分野

職名

  • 教授

学位

  • 博士(学術)(1997年06月 東京大学)

ホームページURL

J-Global ID

研究キーワード

  • 非線形光学   分光学   応用光学   Spectroscopy   Applied Optics   

研究分野

  • ナノテク・材料 / 光工学、光量子科学
  • ナノテク・材料 / 基礎物理化学
  • ライフサイエンス / 生体材料学
  • ライフサイエンス / 生体医工学

職歴

  • 2019年04月 - 現在 北海道大学 大学院情報科学研究院 生命人間情報科学部門 教授
  • 2016年10月 - 2019年03月 北海道大学 大学院情報科学研究科 生命人間情報科学専攻 教授
  • 2007年04月 - 2016年09月 大阪大学 大学院基礎工学研究科 機能創成専攻 生体工学領域 准教授
  • 2003年04月 - 2007年03月 大阪大学 大学院基礎工学研究科 機能創成専攻 生体工学領域 助教授
  • 2002年11月 - 2003年03月 大阪大学 大学院基礎工学研究科 システム人間系専攻 助教授
  • 1999年04月 - 2002年10月 大阪大学 大学院基礎工学研究科 システム人間系専攻 講師
  • 1997年08月 - 1999年03月 大阪大学 大学院基礎工学研究科システム人間系専攻 助手
  • 1996年04月 - 1997年07月 徳島大学工学部機械工学科 助手
  • 1991年04月 - 1996年03月 神奈川科学技術アカデミー 濵口「極限分子計測」プロジェクト 研究員

学歴

  • 1997年06月 -   東京大学   大学院総合文化研究科 論文博士
  • 1989年04月 - 1991年03月   大阪大学   大学院工学研究科   応用物理学専攻
  • 1985年04月 - 1989年03月   大阪大学   工学部   応用物理学科

所属学協会

  • レーザ顕微鏡研究会   日本光学会   日本分光学会   日本応用物理学会   日本機械学会   

研究活動情報

論文

  • Naoki Yamato, Hirohiko Niioka, Jun Miyake, Mamoru Hashimoto
    Scientific Reports 10 1 15212  2020年09月 [査読有り][通常論文]
     
    AbstractA coherent anti-Stokes Raman scattering (CARS) rigid endoscope was developed to visualize peripheral nerves without labeling for nerve-sparing endoscopic surgery. The developed CARS endoscope had a problem with low imaging speed, i.e. low imaging rate. In this study, we demonstrate that noise reduction with deep learning boosts the nerve imaging speed with CARS endoscopy. We employ fine-tuning and ensemble learning and compare deep learning models with three different architectures. In the fine-tuning strategy, deep learning models are pre-trained with CARS microscopy nerve images and retrained with CARS endoscopy nerve images to compensate for the small dataset of CARS endoscopy images. We propose using the equivalent imaging rate (EIR) as a new evaluation metric for quantitatively and directly assessing the imaging rate improvement by deep learning models. The highest EIR of the deep learning model was 7.0 images/min, which was 5 times higher than that of the raw endoscopic image of 1.4 images/min. We believe that the improvement of the nerve imaging speed will open up the possibility of reducing postoperative dysfunction by intraoperative nerve identification.
  • Naoki Yamato, Mana Matsuya, Hirohiko Niioka, Jun Miyake, Mamoru Hashimoto
    Biomolecules 10 7 1012 - 1012 2020年07月08日 [査読有り][通常論文]
     
    Semantic segmentation with deep learning to extract nerves from label-free endoscopic images obtained using coherent anti-Stokes Raman scattering (CARS) for nerve-sparing surgery is described. We developed a CARS rigid endoscope in order to identify the exact location of peripheral nerves in surgery. Myelinated nerves are visualized with a CARS lipid signal in a label-free manner. Because the lipid distribution includes other tissues as well as nerves, nerve segmentation is required to achieve nerve-sparing surgery. We propose using U-Net with a VGG16 encoder as a deep learning model and pre-training with fluorescence images, which visualize the lipid distribution similar to CARS images, before fine-tuning with a small dataset of CARS endoscopy images. For nerve segmentation, we used 24 CARS and 1,818 fluorescence nerve images of three rabbit prostates. We achieved label-free nerve segmentation with a mean accuracy of 0.962 and an F 1 value of 0.860. Pre-training on fluorescence images significantly improved the performance of nerve segmentation in terms of the mean accuracy and F 1 value ( p < 0 . 05 ). Nerve segmentation of label-free endoscopic images will allow for safer endoscopic surgery, while reducing dysfunction and improving prognosis after surgery.
  • Keigo Hirose, Shuichiro Fukushima, Taichi Furukawa, Hirohiko Niioka, Mamoru Hashimoto
    APL Photonics 3 9 092407  2018年07月 [査読有り][招待有り]
  • K. Hirose, T. Aoki, T. Furukawa, S. Fukushima, H. Niioka, S. Deguchi, M. Hashimoto
    Biomedical Optics Express 9 2 387 - 396 2018年02月01日 [査読有り][通常論文]
     
    Label-free visualization of nerves and nervous plexuses will improve the preservation of neurological functions in nerve-sparing robot-assisted surgery. We have developed a coherent anti-Stokes Raman scattering (CARS) rigid endoscope to distinguish nerves from other tissues during surgery. The developed endoscope, which has a tube with a diameter of 12 mm and a length of 270 mm, achieved 0.91% image distortion and 8.6% non-uniformity of CARS intensity in the whole field of view (650 µm diameter). We demonstrated CARS imaging of a rat sciatic nerve and visualization of the fine structure of nerve fibers.
  • Kamel Mars, De Xing Lioe, Shoji Kawahito, Keita Yasutomi, Keiichiro Kagawa, Takahiro Yamada, Mamoru Hashimoto
    SENSORS 17 11 2581  2017年11月 [査読有り][通常論文]
     
    Raman imaging eliminates the need for staining procedures, providing label-free imaging to study biological samples. Recent developments in stimulated Raman scattering (SRS) have achieved fast acquisition speed and hyperspectral imaging. However, there has been a problem of lack of detectors suitable for MHz modulation rate parallel detection, detecting multiple small SRS signals while eliminating extremely strong offset due to direct laser light. In this paper, we present a complementary metal-oxide semiconductor (CMOS) image sensor using high-speed lock-in pixels for stimulated Raman scattering that is capable of obtaining the difference of Stokes-on and Stokes-off signal at modulation frequency of 20 MHz in the pixel before reading out. The generated small SRS signal is extracted and amplified in a pixel using a high-speed and large area lateral electric field charge modulator (LEFM) employing two-step ion implantation and an in-pixel pair of low-pass filter, a sample and hold circuit and a switched capacitor integrator using a fully differential amplifier. A prototype chip is fabricated using 0.11 m CMOS image sensor technology process. SRS spectra and images of stearic acid and 3T3-L1 samples are successfully obtained. The outcomes suggest that hyperspectral and multi-focus SRS imaging at video rate is viable after slight modifications to the pixel architecture and the acquisition system.
  • Takeo Minamikawa, Yoshinori Murakami, Naokazu Matsumura, Hirohiko Niioka, Shuichiro Fukushima, Tsutomu Araki, Mamoru Hashimoto
    JOURNAL OF SPECTROSCOPY 2017 5725340-1 - 5725340-8 2017年 [査読有り][通常論文]
     
    In this study, we investigated photo-induced damage to living cells during single-and multifocus excitations for coherent anti-Stokes Raman scattering (CARS) imaging. A near-infrared pulsed laser (709 nm) was used to induce cell damage. We compared the photo-induced cell damage in the single- and the multifocus excitation schemes with the condition to obtain the same CARS signal in the same frame rate. For the evaluation of cell viability, we employed 4', 6-diamidino-2-phenylindole (DAPI) fluorophores that predominantly stained the damaged cells. One-and two-photon fluorescence of DAPI fluorophores were, respectively, excited by an ultraviolet light source and the same near-infrared light source and were monitored to evaluate the cell viability during near-infrared pulsed laser irradiation. We found lower uptake of DAPI fluorophores into HeLa cells during the multifocus excitation compared with the single- focus excitation scheme in both the one- and the two-photon fluorescence examinations. This indicates a reduction of photo-induced cell damage in the multifocus excitation. Our findings suggested that the multifocus excitation scheme is expected to be suitable for CARS microscopy in terms of minimal invasiveness.
  • Y. Matsuda, J. Miura, M. Shimizu, T. Aoki, M. Kubo, S. Fukushima, M. Hashimoto, F. Takeshige, T. Araki
    JOURNAL OF DENTAL RESEARCH 95 13 1528 - 1534 2016年12月 [査読有り][通常論文]
     
    Advanced glycation end-products (AGEs) are generated via nonenzymatic glycation of dentinal collagen, resulting in accumulation of AGEs in dentin tissue. Since accumulated AGEs cause crosslinking between amino acid polypeptides in the collagen molecule and modify mechanical properties of dentinal collagen, the authors assumed that there would be a significant interaction between the generation of AGEs and progression of caries in dentin. To confirm such an interaction, spectroscopic imaging analyses (i.e., nanosecond fluorescence lifetime imaging and second harmonic generation light imaging) were performed in addition to biochemical and electron microscopic analyses in the present study. Seven carious human teeth were fixed in paraformaldehyde and cut longitudinally into 1-mm sections using a low-speed diamond saw for the following analyses. In transmission electron microscopy (TEM) analysis, nondecalcified specimens were embedded in epoxy resin and sliced into thin sections for observation. For the immunohistochemical analysis, the specimens were paraffin embedded after decalcification for 2 wk and sectioned with a microtome. Resultant sections were stained with anti-AGE and anticollagen antibodies. The demineralized specimens were used for spectroscopic analyses without additional treatment. For Western blotting analysis, specimens were separated into carious and sound dentin. Each specimen was homogenized with a bead crusher and an ultrasonic homogenizer and then treated with hydrochloric acid. In carious dentin, the collagen fibers showed an amorphous structure in the TEM image, and the AGEs were localized in the areas of bacterial invasion in the immunostaining image. The total amount of AGEs in carious dentin was higher than in sound dentin in Western blotting. The ultrastructure of type I collagen and total amount of AGEs varied markedly in the dentinal caries region. The fluorescence lifetime was shorter in the carious area than that in the sound areas, indicating an increase of AGEs in the carious area. The increase of AGEs could influence the progression of dentinal caries.
  • Doan Thi Kim Dung, Shoichiro Fukushima, Taichi Furukawa, Hirohiko Niioka, Takumi Sannomiya, Kaori Kobayashi, Hiroshi Yukawa, Yoshinobu Baba, Mamoru Hashimoto, Jun Miyake
    NANOMATERIALS 6 9 163-1 - 163-17 2016年09月 [査読有り][通常論文]
     
    Comprehensive imaging of a biological individual can be achieved by utilizing the variation in spatial resolution, the scale of cathodoluminescence (CL), and near-infrared (NIR), as favored by imaging probe Gd2O3 co-doped lanthanide nanophosphors (NPPs). A series of Gd2O3:Ln(3+)/Yb3+ (Ln(3+): Tm3+, Ho3+, Er3+) NPPs with multispectral emission are prepared by the sol-gel method. The NPPs show a wide range of emissions spanning from the visible to the NIR region under 980 nm excitation. The dependence of the upconverting (UC)/downconverting (DC) emission intensity on the dopant ratio is investigated. The optimum ratios of dopants obtained for emissions in the NIR regions at 810 nm, 1200 nm, and 1530 nm are applied to produce nanoparticles by the homogeneous precipitation (HP) method. The nanoparticles produced from the HP method are used to investigate the dual NIR and CL imaging modalities. The results indicate the possibility of using Gd2O3 co-doped Ln(3+)/Yb3+ (Ln(3+): Tm3+, Ho3+, Er3+) in correlation with NIR and CL imaging. The use of Gd2O3 promises an extension of the object dimension to the whole-body level by employing magnetic resonance imaging (MRI).
  • S. Fukushima, T. Furukawa, H. Niioka, M. Ichimiya, T. Sannomiya, N. Tanaka, D. Onoshima, H. Yukawa, Y. Baba, M. Ashida, J. Miyake, T. Araki, M. Hashimoto
    SCIENTIFIC REPORTS 6 25950  2016年05月 [査読有り][通常論文]
     
    This paper presents a new correlative bioimaging technique using Y2O3:Tm, Yb and Y2O3:Er, Yb nanophosphors (NPs) as imaging probes that emit luminescence excited by both near-infrared (NIR) light and an electron beam. Under 980 nm NIR light irradiation, the Y2O3:Tm, Yb and Y2O3:Er, Yb NPs emitted NIR luminescence (NIRL) around 810 nm and 1530 nm, respectively, and cathodoluminescence at 455 nm and 660 nm under excitation of accelerated electrons, respectively. Multimodalities of the NPs were confirmed in correlative NIRL/CL imaging and their locations were visualized at the same observation area in both NIRL and CL images. Using CL microscopy, the NPs were visualized at the single-particle level and with multicolour. Multiscale NIRL/CL bioimaging was demonstrated through in vivo and in vitro NIRL deep-tissue observations, cellular NIRL imaging, and high-spatial resolution CL imaging of the NPs inside cells. The location of a cell sheet transplanted onto the back muscle fascia of a hairy rat was visualized through NIRL imaging of the Y2O3:Er, Yb NPs. Accurate positions of cells through the thickness (1.5 mm) of a tissue phantom were detected by NIRL from the Y2O3:Tm, Yb NPs. Further, locations of the two types of NPs inside cells were observed using CL microscopy.
  • De Xing Lioe, Kamel Mars, Shoji Kawahito, Keita Yasutomi, Keiichiro Kagawa, Takahiro Yamada, Mamoru Hashimoto
    SENSORS 16 4 532-1 - 532-16 2016年04月 [査読有り][通常論文]
     
    A complementary metal-oxide semiconductor (CMOS) lock-in pixel to observe stimulated Raman scattering (SRS) using a high speed lateral electric field modulator (LEFM) for photo-generated charges and in-pixel readout circuits is presented. An effective SRS signal generated after the SRS process is very small and needs to be extracted from an extremely large offset due to a probing laser signal. In order to suppress the offset components while amplifying high-frequency modulated small SRS signal components, the lock-in pixel uses a high-speed LEFM for demodulating the SRS signal, resistor-capacitor low-pass filter (RC-LPF) and switched-capacitor (SC) integrator with a fully CMOS differential amplifier. AC (modulated) components remained in the RC-LPF outputs are eliminated by the phase-adjusted sampling with the SC integrator and the demodulated DC (unmodulated) components due to the SRS signal are integrated over many samples in the SC integrator. In order to suppress further the residual offset and the low frequency noise (1/f noise) components, a double modulation technique is introduced in the SRS signal measurements, where the phase of high-frequency modulated laser beam before irradiation of a specimen is modulated at an intermediate frequency and the demodulation is done at the lock-in pixel output. A prototype chip for characterizing the SRS lock-in pixel is implemented and a successful operation is demonstrated. The reduction effects of residual offset and 1/f noise components are confirmed by the measurements. A ratio of the detected small SRS to offset a signal of less than 10(-5) is experimentally demonstrated, and the SRS spectrum of a Benzonitrile sample is successfully observed.
  • Shoichiro Fukushima, Taichi Furukawa, Hirohiko Niioka, Masayoshi Ichimiya, Takumi Sannomiya, Jun Miyake, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto
    OPTICAL MATERIALS EXPRESS 6 3 831 - 843 2016年03月 [査読有り][通常論文]
     
    Yttrium oxide-based nanophosphors that emit both upconversion luminescence (UPL) and cathodoluminescence (CL) were synthesized by a precipitation method using excessive urea. Precursors of Y2O3 nanophosphors were synthesized with size control to less than 50 nm and a chemical yield greater than 90%. Concentrations of rare-earth co-dopants in nanophosphors were controlled with optimal molar ratios. Co-dopants Tm, Yb/Er, Yb enabled NPs to emit UPL at wavelengths around 810/660 nm and CL at wavelengths around 450/660 nm via excitation with 980 nm NIR light and an electron beam. Synthesized NPs were imaged by NIR and CL microscopy. (C) 2016 Optical Society of America
  • DeXing Lioe, Kamel Mars, Taishi Takasawa, Keita Yasutomi, Keiichiro Kagawa, Mamoru Hashimoto, Shoji Kawahito
    HIGH-SPEED BIOMEDICAL IMAGING AND SPECTROSCOPY: TOWARD BIG DATA INSTRUMENTATION AND MANAGEMENT 9720 97200J  2016年 [査読有り][通常論文]
     
    A CMOS image sensor using high-speed lock-in pixels for stimulated Raman scattering (SRS) spectroscopy is presented in this paper. The effective SRS signal from the stimulated emission of SRS mechanism is very small in contrast to the offset of a probing laser source, which is in the ratio of 10(-4) to 10(-5). In order to extract this signal, the common offset component is removed, and the small difference component is sampled using switched-capacitor integrator with a fully differential amplifier. The sampling is performed over many integration cycles to achieve appropriate amplification. The lock-in pixels utilizes high-speed lateral electric field charge modulator (LEFM) to demodulate the SRS signal which is modulated at high-frequency of 20MHz. A prototype chip is implemented using 0.11 mu m CMOS image sensor technology.
  • Mamoru Hashimoto
    IEEE CPMT Symposium Japan 2015: Packaging is Everywhere, ICSJ 2015 88 - 89 2015年12月14日 [査読無し][通常論文]
     
    Raman imaging visualizes the biomolecule without staining however, the low cross section of Raman scattering requires long observation time. We have developed a nonlinear Raman microscope and endoscope for fast biomolecular imaging without staining. In the presentation, I would like to talk about the present status of nonlinear Raman imaging and medical applications.
  • Sho Okubo, Yi-Da Hsieh, Hajime Inaba, Atsushi Onae, Mamoru Hashimoto, Takeshi Yasui
    OPTICS EXPRESS 23 26 33184 - 33193 2015年12月 [査読有り][通常論文]
     
    We performed broadband dual-frequency-comb spectroscopy in the near-infrared region with a much higher resolution than the Fourier limit by using discrete Fourier transforms and spectral interleaving. We observed the resonant spectrum of a Fabry-Perot cavity over a spectral range of 187 to 218 THz using this technique, and measured its free spectral ranges and finesses. The recorded spectrum includes cavity resonance lines with widths of about 2 MHz, which is much narrower than the resolution of 48 MHz determined by the observation time window. (C) 2015 Optical Society of America
  • Mamoru Hashimoto, Keisuke Yoshiki, Makoto Kurihara, Nobuyuki Hashimoto, Tsutomu Araki
    OPTICAL REVIEW 22 6 875 - 881 2015年12月 [査読有り][通常論文]
     
    We have developed a system for measuring the orientation of single molecules using a conventional wide-field fluorescence microscope with a polarization filter consisting of a polarizer and a compact polarization mode converter. The polarization filter electrically controls the pattern of polarization filtering. Since the polarization of the fluorescence from a single molecule highly depends on the angle between the observation direction and the molecular direction, polarization pattern filtering at the pupil plane of the objective lens allows the orientation of a single molecule to be visualized. Using this system, we demonstrated the orientation detection of single molecules.
  • Mamoru Hashimoto, Hirohiko Niioka, Koichiro Ashida, Keisuke Yoshiki, Tsutomu Araki
    APPLIED PHYSICS EXPRESS 8 11 112401  2015年11月 [査読有り][通常論文]
     
    High-sensitivity, high-spatial-resolution imaging of organic monolayers on platinum with second harmonic generation (SHG) microscopy using radially polarized beam excitation is investigated. A tightly focused, radially polarized beam forms a longitudinal electric field at the focus. The longitudinal field is enhanced at a metal surface and increases the intensity of SHG from the molecules on the metal surface. The SHG signal from a self-assembled monolayer (SAM) on a platinum surface excited by a radially polarized beam is approximately 3.7 times higher than that obtained with a linearly polarized beam. Improved spatial resolution is also demonstrated using a SAM patterned by electron beam lithography. (C) 2015 The Japan Society of Applied Physics
  • Takeshi Yasui, Yuki Iyonaga, Yi-Da Hsieh, Yoshiyuki Sakaguchi, Francis Hindle, Shuko Yokoyama, Tsutomu Araki, Mamoru Hashimoto
    Optica 2 5 460 - 460 2015年05月20日 [査読有り][通常論文]
     
    Fourier transform spectroscopy (FTS) has been widely used in a variety of fields due to its high signal-to-noise ratio, simultaneous acquisition of a broad spectrum, and versatility for different radiation sources. Further improvement of the spectroscopic performance will widen its scope of applications. Here, we demonstrate improved spectral resolution by overcoming the time-window size limitation using a mode-locked terahertz (THz) pulse train as precisely periodic pulsed radiation in discrete Fourier transform spectroscopy (dFTS). Since infinitesimal resolution can be achieved at harmonic components of its repetition frequency 1/𝑇 when the time-window size is exactly matched to the repetition period 𝑇, a combination of dFTS with a spectral interleaving technique achieves a spectral resolution limited only by the spectral interleaving interval. Linewidths narrower than 1/(50𝑇) are fully resolved by THz-dFTS, allowing rotational-transition absorption lines of low-pressure molecular gases to be attributed within a 1.25 MHz band.
  • Taichi Furukawa, Shoichiro Fukushima, Hirohiko Niioka, Naoki Yamamoto, Jun Miyake, Tsutomu Araki, Mamoru Hashimoto
    JOURNAL OF BIOMEDICAL OPTICS 20 5 056007-1 - 056007-6 2015年05月 [査読有り][通常論文]
     
    We describe rare-earth-doped nanophosphors (RE-NPs) for biological imaging using cathodoluminescence (CL) microscopy based on scanning transmission electron microscopy (STEM). We report the first demonstration of multicolor CL nanobioimaging using STEM with nanophosphors. The CL spectra of the synthesized nanophosphors (Y2O3:Eu, Y2O3:Tb) were sufficiently narrow to be distinguished. From CL images of RE-NPs on an elastic carbon-coated copper grid, the spatial resolution was beyond the diffraction limit of light. Y2O3:Tb and Y2O3:Eu RE-NPs showed a remarkable resistance against electron beam exposure even at high acceleration voltage (80 kV) and retained a CL intensity of more than 97% compared with the initial intensity for 1 min. In biological CL imaging with STEM, heavy-metal-stained cell sections containing the RE-NPs were prepared, and both the CL images of RE-NPs and cellular structures, such as mitochondria, were clearly observed from STEM images with high contrast. The cellular CL imaging using RE-NPs also had high spatial resolution even though heavy-metal-stained cells are normally regarded as highly scattering media. Moreover, since the RE-NPs exhibit photoluminescence (PL) excited by UV light, they are useful for multimodal correlative imaging using CL and PL. (C) 2015 Society of Photo-Optical Instrumentation Engineers (SPIE)
  • Shuichiro Fukushima, Masato Shimizu, Jiro Miura, Yusuke Matsuda, Mizuho Kubo, Mamoru Hashimoto, Takuya Aoki, Fumio Takeshige, Tsutomu Araki
    BIOMEDICAL OPTICS EXPRESS 6 5 1844 - 1856 2015年05月 [査読有り][通常論文]
     
    Advanced Glycation End-products (AGEs) are produced by the Maillard reaction, which causes cross-linking of collagen and results in changes in the mechanical properties of collagen tissues. Several types of AGE fluoresce, and measurement of this fluorescence is effective for determining the presence of AGEs. Because fluorescence intensity by steady-state fluorometry is affected by sample surface condition and light source, we focused on fluorescence lifetime measurement (FLM). We found that fluorescence lifetime of collagen gel decreased with glycation progress. In vivo application of FLM for determination of AGEs was confirmed in human dentin. (C) 2015 Optical Society of America
  • Y. -D. Hiseh, S. Okubo, H. Inaba, M. Hashimoto, T. Yasui
    2015 CONFERENCE ON LASERS AND ELECTRO-OPTICS (CLEO) 2015年 [査読有り][通常論文]
     
    A stabilized Fabry-Perot cavity with a 2.2 MHz-linewidth resonance-mode and a 566MHz FSR was fully characterized over the spectral range from 186 to 200 THz by discrete Fourier transform infrared spectroscopy using precisely periodic pulse.
  • Kim Dung T. Doan, Shoichiro Fukushima, Hirohiko Niioka, Masayoshi Ichimiya, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto, Jun Miyake
    BIOPHYSICAL JOURNAL 108 2 171A - 172A 2015年01月 [査読有り][通常論文]
  • Shoichiro Fukushima, Taichi Furukawa, Hirohiko Niioka, Masayoshi Ichimiya, Jun Miyake, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto
    MICRON 67 90 - 95 2014年12月 [査読有り][通常論文]
     
    We present a phosphor nanoparticle that shows both upconversion luminescence (UCL) and cathodoluminescence (CL). With this particle, low-autofluorescence, deep-tissue and wide-field fluorescence imaging can be achieved with nanometer-order high-spatial-resolution imaging. We synthesized Y2O3:Tm,Yb nanophosphors that emit visible and near-infrared UCL under 980 nm irradiation and blue CL via electron beam excitation. The phosphors were applied to fluorescent imaging of HeLa cells. The photostability of the phosphors was superior to that of a conventional organic dye. We show that after uptake by HeLa cells, the particles can be imaged with SEM and CL contrast in a cellular section. This indicates that correlative UCL and CL imaging of biological samples could be realized. (C) 2014 Elsevier Ltd. All rights reserved.
  • Jiro Miura, Kantaro Nishikawa, Mizuho Kubo, Shuichiro Fukushima, Mamoru Hashimoto, Fumio Takeshige, Tsutomu Araki
    ARCHIVES OF ORAL BIOLOGY 59 2 119 - 124 2014年02月 [査読有り][通常論文]
     
    Cross-linking of collagen by Advanced Glycation End-products (AGEs) occurs by nonenzymatic glycation (Maillard reaction). The purpose of this study was to examine whether AGEs are formed in human dentinal collagen, and to consider any possible influence of AGEs on dentinal physiology. Mechanical characteristics, fluorescence spectra and immunohistochemical analyses of demineralized dentine sections from young subjects were compared with those of aged ones. The same investigations were performed with young dentine artificially glycated by incubation in 0.1 M ribose solution. Indentation measurement indicated that the sections from aged dentine were mechanically harder than those from young dentine. The hardness of young dentine increased after incubation in ribose solution. Fluorescence peak wavelength of the young dentine was shorter than that of the aged one, but shifted towards the peak wavelength of the aged one after incubation in ribose solution. These changes were considered to be due to accumulation of AGEs. Existence of AGEs in dentinal collagen was confirmed by immunohistochemical analysis. The obtained results suggest that AGEs accumulation occurs in dentinal collagen and is affected by both human age and physiological conditions such as glucose level in blood because dentinal collagen receives nourishment via dental pulp and tubules. (C) 2013 Elsevier Ltd. All rights reserved.
  • Mamoru Hashimoto, Hiroto Kanoh, Hirohiko Niioka, Tsutomu Araki
    Progress in Biomedical Optics and Imaging - Proceedings of SPIE 8948 2014年 [査読有り][通常論文]
     
    Coherent Raman scattering provides chemical imaging by using molecular vibrational information sensitive to molecular structure. To add another information of martial symmetry, we propose using fourth order coherent Raman scattering for imaging, because the even order nonlinear phenomenon is forbidden for centro-symmetric material. We have developed a multiplex fourth order coherent Raman scattering microscopy system using a femtosecond laser. A narrowband beam of 17 cm-1 bandwidth and a broadband beam generated by a photonic crystal fiber enables to obtain a spectrum of fourth order coherent Raman scattering at once. We demonstrate the fourth order coherent Raman, hyper-Raman and second harmonics of trans-4'-(dimethylamino)-N-methyl-4- stilbazolium tosylate crystal by using the developed microscope. © 2014 SPIE.
  • Kamel Mars, Ken Egawa, Lioe De Xing, Han Sang Man, Taishi Takasawa, Keita Yasutomi, Keiichiro Kagawa, Mamoru Hashimoto, Shoji Kawahito
    2014 IEEE 12TH INTERNATIONAL NEW CIRCUITS AND SYSTEMS CONFERENCE (NEWCAS) 4238 9 249 - 252 2014年 [査読有り][通常論文]
     
    In this paper, a lock-in pixel readout circuits using a high speed lateral electric field modulator with differential charge accumulation for a stimulated Raman scattering imager is presented. The stimulated Raman scattering works by detecting the vibrations in chemical bonds between atoms, and the generated Raman signal after stimulated scattering processes is very low compared to offset signal. By using a high-speed lateral electric field modulator with lock-in pixels differential charge accumulation technique using a sample-and hold circuit with a fully differential amplifier, very small effective Raman signal can be extracted from large offset signal and high dynamic range is achieved.
  • Kamel Mars, Beak Guseul, Han Sang Man, Taishi Takasawa, Keita Yasutomi, Keiichiro Kagawa, Mamoru Hashimoto, Shoji Kawahito
    IMAGE SENSORS AND IMAGING SYSTEMS 2014 9022 90220D  2014年 [査読有り][通常論文]
     
    A high speed Lateral Electric Field Modulator (LEFM) and lock-in pixels amplifiers for stimulated Raman scattering (SRS) imager is presented. Since the generated signal from the SRS process is very small compared to the offset signal, a technique suitable for extracting and amplifying the SRS signal is needed. The offset can be canceled by tuning the phase delay between the demodulated pixel output signal and the sampling clock. The small SRS signal in large offset is amplified by the differential integration. The proposed technique has been investigated with an implementation of 64x8 pixels array using a pinned photodiode LEFM an lock-in pixels amplifiers. Very small signal can be extracted from large offset signal. A ratio of the detected small SRS to offset signal of less 10(-5) is achieved.
  • Mamoru Hashimoto, Hiroto Kanoh, Hirohika Niioka, Tsutomu Araki
    MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES XIV 8948 894817  2014年 [査読有り][通常論文]
     
    Coherent Raman scattering provides chemical imaging by using molecular vibrational information sensitive to molecular structure. To add another information of martial symmetry, we propose using fourth order coherent Raman scattering for imaging, because the even order nonlinear phenomenon is forbidden for centro-symmetric material. We have developed a multiplex fourth order coherent Raman scattering microscopy system using a femtosecond laser. A narrowband beam of 17 cm(-1) bandwidth and a broadband beam generated by a photonic crystal fiber enables to obtain a spectrum of fourth order coherent Raman scattering at once. We demonstrate the fourth order coherent Raman, hyper-Raman and second harmonics of trans-4'-(dimethylamino)-N-methyl-4-stilbazolium tosylate crystal by using the developed microscope.
  • Taichi Furukawa, Hirohiko Niioka, Masayoshi Ichimiya, Tomohiro Nagata, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto
    Optics Express 21 22 25655 - 25663 2013年11月04日 [査読有り][通常論文]
     
    High-resolution microscopy for biological specimens was performed using cathodoluminescence (CL) of Y2O3:Eu, Zn nanophosphors, which have high CL intensity due to the incorporation of Zn. The intensity of Y2O3:Eu nanophosphors at low acceleration voltage (3 kV) was increased by adding Zn. The CL intensity was high enough for imaging even with a phosphor size as small as about 30 nm. The results show the possibility of using CL microscopy for biological specimens at single-protein-scale resolution. CL imaging of HeLa cells containing laserablated Y2O 3:Eu, Zn nanophosphors achieved a spatial resolution of a few tens of nanometers. Y2O3:Eu, Zn nanophosphors in HeLa cells were also imaged with 254 nm ultraviolet light excitation. The results suggest that correlative microscopy using CL, secondary electrons and fluorescence imaging could enable multi-scale investigation of molecular localization from the nanoscale to the microscale. ©2013 Optical Society of America.
  • Harsono Cahyadi, Junichi Iwatsuka, Takeo Minamikawa, Hirohiko Niioka, Tsutomu Araki, Mamoru Hashimoto
    JOURNAL OF BIOMEDICAL OPTICS 18 9 096009  2013年09月 [査読有り][通常論文]
     
    We develop a coherent anti-Stokes Raman scattering (CARS) microscopy system equipped with a tunable picosecond laser for high-speed wavelength scanning. An acousto-optic tunable filter (AOTF) is integrated in the laser cavity to enable wavelength scanning by varying the radio frequency waves applied to the AOTF crystal. An end mirror attached on a piezoelectric actuator and a pair of parallel plates driven by galvanometer motors are also introduced into the cavity to compensate for changes in the cavity length during wavelength scanning to allow synchronization with another picosecond laser. We demonstrate fast spectral imaging of 3T3-L1 adipocytes every 5 cm(-1) in the Raman spectral region around 2850 cm(-1) with an image acquisition time of 120 ms. We also demonstrate fast switching of Raman shifts between 2100 and 2850 cm(-1), corresponding to CD2 symmetric stretching and CH2 symmetric stretching vibrations, respectively. The fast-switching CARS images reveal different locations of recrystallized deuterated and nondeuterated stearic acid. (C) The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
  • Takeo Minamikawa, Tatsuro Takagi, Hirohiko Niioka, Makoto Kurihara, Nobuyuki Hashimoto, Tsutomu Araki, Mamoru Hashimoto
    APPLIED PHYSICS EXPRESS 6 7 072401  2013年07月 [査読有り][通常論文]
     
    We have developed a tunable-polarization-mode coherent anti-Stokes Raman scattering (CARS) microscope with compact polarization mode converters constructed using eight-segmented liquid-crystal spatial light modulators. The polarization modes, such as linear, radial, and azimuthal polarizations, of two excitation beams are controlled independently and are switched without any mechanical tuning in less than 300 ms. We use the system to detect the molecular orientation of 4-cyano-4'-octylbiphenyl (8CB) liquid crystals aligned parallel and perpendicular to the optical axis. We also observe CARS images of liquid crystal defects known as focal conic domains, demonstrating the potential of our molecular orientation imaging system. (C) 2013 The Japan Society of Applied Physics
  • Mamoru Hashimoto, Takeo Minamikawa, Tsutomu Araki
    CURRENT PHARMACEUTICAL BIOTECHNOLOGY 14 2 150 - 158 2013年02月 [査読有り][招待有り]
     
    Vibrational microscopy (Raman microscopy and infrared microscopy), which observes molecular vibrations, gives us the information of molecular species without staining because the observed signals are originated from intrinsic molecules of a cell. However, infrared radiation is absorbed with water, and the long wavelength (3-10 mu m) limits the spatial resolution to several micrometers. Spontaneous emission of Raman scattering is quite feeble, and the Raman scattering often overlaps with one-photon fluorescence from a specimen. Coherent anti-Stokes Raman scattering (CARS) microscopy, which is one of the nonlinear Raman microscopy, is a method to overcome those problems. In this review, present system of CARS microscopy, the methods of background rejection, and applications are introduced.
  • Zhuo Li, Shoji Kawahito, Keita Yasutomi, Keiichiro Kagawa, Juichiro Ukon, Mamoru Hashimoto, Hirohiko Niioka
    IEEE TRANSACTIONS ON ELECTRON DEVICES 59 10 2715 - 2722 2012年10月 [査読有り][通常論文]
     
    This paper presents a time-resolved CMOS image sensor with draining-only modulation (DOM) pixels, for time-domain fluorescence lifetime imaging. In the DOM pixels using a pinned photodiode (PPD) technology, a time-windowed signal charge transfer from a PPD to a pinned storage diode (PSD) is controlled by a draining gate only, without a transfer gate between the two diodes. This structure allows a potential barrierless and trapless charge transfer from the PPD to the PSD. A 256 x 256 pixel time-resolved CMOS imager with 7.5 x 7.5 mu m(2) DOM pixels has been implemented using 0.18-mu m CMOS image sensor process technology with PPD option. The prototype demonstrates high sensitivity for weak signal of less than one electron per light pulse and accurate measurement of fluorescence decay process with subnanosecond time resolution.
  • Mamoru Hashimoto, Taro Ichimura, Katsumasa Fujita
    Springer Series in Optical Sciences 168 317 - 346 2012年 [査読有り][通常論文]
     
    Raman microscopy has been attracting researchers in biology and medicine due to its capability of detecting molecular vibrations that provide information of molecular species, structures, conditions, and environments. Raman scattering can be obtained by simply illuminating molecules with monochromatic light, and providing molecular vibration frequency as wavelengths of scattered light. This does not require labeling of target molecules, such as chemical or biological staining with fluorophore, which may modify the condition of living biological specimens. © 2012 Springer-Verlag Berlin Heidelberg.
  • Mamoru Hashimoto, Junichi Iwatsuka, Hirohiko Niioka, Tsutomu Araki
    Progress in Biomedical Optics and Imaging - Proceedings of SPIE 8226 2012年 [査読有り][通常論文]
     
    We have developed a high speed spectral tuning CARS microscopy system using a mode-locked Ti:Sapphire laser with an acousto-optic tunable filter (AOTF) in the cavity. Since the wavelength of the laser is tunable with the applied radio frequency to the AOTF, the wavelength is electrically tunable.The pulse duration of the laser is about 10 ps, tunable range is 800 nm to 930 nm, and the tuning speed is ms order. The laser is synchronized with another mode-locked Ti:Sapphire laser laser our own method using a balance cross-correlator and phase lock loop technique. The synchronized lasers are used for light source of multi-focus CARS microscopy system using a microlens array scanner, and the hyperspectral imaging of adipocyte cells is demonstrated. © 2012 Copyright Society of Photo-Optical Instrumentation Engineers (SPIE).
  • Hirohiko Niioka, Taichi Furukawa, Masayoshi Ichimiya, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto
    APPLIED PHYSICS EXPRESS 4 11 2011年11月 [査読有り][通常論文]
     
    We report the first demonstration of a multicolor high-spatial-resolution imaging technique for observation of biological cells using cathodoluminescence from nanophosphors. Three kinds of rare-earth-doped nanophosphors were injected into J744A.1 macrophages, and the spatial distribution of nanophosphors was visualized by using a scanning electron microscope cathodoluminescence (SEM-CL) system. The spectral bandwidth of the phosphors was narrow enough to distinguish the types of the phosphors. CL images of the nanophosphors on Si substrates were obtained with high resolution comparable to that of SEM images. These nanophosphors will be candidates to image more than two kinds of biological molecules at high resolution. (C) 2011 The Japan Society of Applied Physics
  • Takeo Minamikawa, Hirohiko Niioka, Tsutomu Araki, Mamoru Hashimoto
    JOURNAL OF BIOMEDICAL OPTICS 16 2 021111  2011年02月 [査読有り][通常論文]
     
    We demonstrate the real-time imaging of laser-induced disruption of the cellular membrane in a living HeLa cell and its cellular response with a multifocus coherent anti-Stokes Raman scattering (CARS) microscope. A near-infrared pulsed laser beam tightly focused on the cellular membrane of a living cell induces ablation at the focal point causing a local disruption of the cellular membrane. After the membrane disruption a dark spot decreasing CARS intensity of 2840 cm(-1) Raman shift at the disrupted site appears. This dark spot immediately disappears and a strong CARS signal is observed around the disrupted site. This increase of the CARS signal might be caused by resealing of the disrupted site via aggregation of the patch lipid vesicles in the cytoplasm. The accumulation of lipids around the disrupted site is also confirmed with three-dimensional CARS images of a cell before and after membrane disruption. The temporal behavior of the CARS signal at the disrupted site is observed to detect the fusion dynamics of patch vesicles. (C) 2011 Society of Photo-Optical Instrumentation Engineers (SPIE). [DOI:10.1117/1.3533314]
  • Mamoru Hashimoto, Tatsuro Takagi, Takeo Minamikawa, Hirohiko Niioka, Tsutomu Araki
    Progress in Biomedical Optics and Imaging - Proceedings of SPIE 7903 2011年 [査読有り][通常論文]
     
    We developed a polarization-mode controllable coherent anti-Stokes Raman scattering microscope. The polarization-mode of excitations beams such as linear, radial, or azimuth polarization were switched with compact polarization mode converters made of eight-segmented liquid-crystal spatial-light-modulators. The polarization-mode of the excitation beams is electrically controllable without any mechanical operation. We demonstrated the detection of the molecular orientation of liquid crystals with the developed microscope. © 2011 SPIE.
  • Takeo Minamikawa, Yoshinori Murakami, Naokazu Matsumura, Hirohiko Niioka, Shuichiro Fukushima, Tsutomu Araki, Mamoru Hashimoto
    Progress in Biomedical Optics and Imaging - Proceedings of SPIE 7903 2011年 [査読有り][通常論文]
     
    We investigated photo-induced cell damage for multi-focus CARS (coherent anti-Stokes Raman scattering) microscopy. In general, using a near-infrared pulse light source, photo-induced damage is dominantly caused via multi-photon induced phenomena, and the peak power of the excitation light is limited for the non-invasive imaging. We obtained cell viability images during single- or multi-focus (7 foci) exposure of which wavelength and pulse duration were 709 nm and 5 ps. The laser power of one focal spot was respectively set to 27.8 mW and 14.5 mW for single- and multi-focus excitation because those excitation beams induce the comparable signals for third-order nonlinear phenomena. The cell viability was observed using DAPI fluorophore that mainly stains DNA of dead cells. As a result, we found that the single-focus excitation with 27.8 mW/spot caused cell damage within 6 min. In contrast, photo-induced damage was not detected until 20 min for the multi-focus excitation with 14.5 mW/spot and 7 foci. The results suggest that the photo-induced damage is a serious problem on the single-focus excitation, and the multi-focus excitation method is preferable for CARS imaging. © 2011 SPIE.
  • K. Yoshiki, S. Yoshida, T. Namazu, N. Araki, M. Hashimoto, M. Kurihara, N. Hashimoto, S. Inoue
    Proceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS) 461 - 464 2011年 [査読有り][通常論文]
     
    In this study we describe a novel system for measuring the distribution of inhomogeneous mechanical properties of microelectromechanical system (MEMS) devices. Such inhomogeneous properties are important because they can decrease the reliability of MEMS devices. Our technique involves measuring the variation in the relative position and angle between single molecule markers sprayed on a MEMS device, which indicates the amount of local deformation. The distribution of the deformation on the surface of a MEMS device is calculated from local displacements measured using the system. To simultaneously measure the three-dimensional (3D) position and orientation of the markers, we developed a 3D orientation measurement system that consists of an epifluorescence microscope and a polarization mode converter (PMC). We also detected the 3D displacement and orientation of the torsion bar in a MEMS mirror device.
  • Takeo Minamikawa, Yoshinori Murakami, Naokazu Matsumura, Hirohiko Niioka, Shuichiro Fukushima, Tsutomu Araki, Mamoru Hashimoto
    MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES XI 7903 79032H  2011年 [査読有り][通常論文]
     
    We investigated photo-induced cell damage for multi-focus CARS (coherent anti-Stokes Raman scattering) microscopy. In general, using a near-infrared pulse light source, photo-induced damage is dominantly caused via multi-photon induced phenomena, and the peak power of the excitation light is limited for the non-invasive imaging. We obtained cell viability images during single- or multi-focus (7 foci) exposure of which wavelength and pulse duration were 709 nm and 5 ps. The laser power of one focal spot was respectively set to 27.8 mW and 14.5 mW for single- and multi-focus excitation because those excitation beams induce the comparable signals for third-order nonlinear phenomena. The cell viability was observed using DAPI fluorophore that mainly stains DNA of dead cells. As a result, we found that the single-focus excitation with 27.8 mW/spot caused cell damage within 6 min. In contrast, photo-induced damage was not detected until 20 min for the multi-focus excitation with 14.5 mW/spot and 7 foci. The results suggest that the photo-induced damage is a serious problem on the single-focus excitation, and the multi-focus excitation method is preferable for CARS imaging.
  • Mamoru Hashimoto, Tatsuro Takagi, Takeo Minamikawa, Hirohiko Niioka, Tsutomu Araki
    MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES XI 7903 79031E  2011年 [査読有り][通常論文]
     
    We developed a polarization-mode controllable coherent anti-Stokes Raman scattering microscope. The polarization-mode of excitations beams such as linear, radial, or azimuth polarization were switched with compact polarization mode converters made of eight-segmented liquid-crystal spatial-light-modulators. The polarization-mode of the excitation beams is electrically controllable without any mechanical operation. We demonstrated the detection of the molecular orientation of liquid crystals with the developed microscope.
  • Shintaro Yoshida, Keisuke Yoshiki, Takahiro Namazu, Nozomu Araki, Mamoru Hashimoto, Makoto Kurihara, Nobuyuki Hashimoto, Shozo Inoue
    OPTICS AND PHOTONICS FOR INFORMATION PROCESSING V 8134 81340E  2011年 [査読有り][通常論文]
     
    We propose a technique that employs single fluorescent molecules for visualizing the distribution of strain induced in microstructures. We sprayed single-molecule tracers on microstructures by ultrasonic atomization and traced the position and orientation of the tracers by a single-molecule detection technique with a three-dimensional (3D) orientation microscope, which consists of a conventional fluorescent microscope and a polarization-mode converter. By using 3D spline interpolation, we visualized the surface geometry of a microelectromechanical (MEMS) device. We tracked the 3D position and orientation of tracers attached to a supporting beam of the MEMS mirror. The surface declination angles calculated from the orientation of the tracers were in agreement with the tilt angle obtained from the 3D position of the tracers.
  • 橋本 守, 南川 丈夫, 新岡 宏彦, 荒木 勉
    日本レーザー医学会誌 = The Journal of Japan Society for Laser Medicine 30 4 421 - 426 特定非営利活動法人 日本レーザー医学会 2010年01月30日 [査読無し][通常論文]
     
    ラマン分光顕微鏡は,全ての分子が持つ分子振動を観測することで,無染色で分子種を見分けながら可視化する手法である.我々は,非線形なラマン散乱分光であるCARS(Coherent anti-Stokes Raman scattering)分光により,リアルタイムで観測可能なを多焦点CARS 顕微鏡を開発した.本稿では,開発した装置や,これを用いて観測した細胞の3次元イメージ,レーザーアブレーションと組み合わせて,細胞内小器官の破壊や細胞膜の破壊と修復過程の観測に応用した例を紹介する.
  • Mamoru Hashimoto, Takeo Minamikawa, Tsutomu Araki
    Progress in Biomedical Optics and Imaging - Proceedings of SPIE 7569 2010年 [査読有り][通常論文]
     
    We developed a high speed CARS (coherent anti-Stokes Raman scattering) spectral-imaging system using an acousto optic tunable filter and multi-focus excitation system. We compared two methods of CARS emission filtering and CARS excitation filtering. In both case, two laser pulses with narrow band (picosecond laser) and broad band (femotosecond laser)were used for the light source. For CARS emission filtering, the generated CARS was filtered by an AOTF, and for excitation filtering the broad band femtosecond laser pulse were filtered by an AOTF before excitation. The experimental results indicated that the CARS emission filtering was suitable for CARS microscopy. © 2010 Copyright SPIE - The International Society for Optical Engineering.
  • Takeo Minamikawa, Tsutomu Araki, Mamoru Hashimoto
    Progress in Biomedical Optics and Imaging - Proceedings of SPIE 7569 2010年 [査読有り][通常論文]
     
    We demonstrated real-time imaging of organelles in a living HeLa cell using a multi-focus excitation CARS (coherent anti-Stokes Raman scattering) microscope. Chemical selective CARS imaging of lipids and proteins was demonstrated by observing CH2 and CH3 vibrations. Real-time imaging of lipid rich organelles such as the plasma membrane, mitochondria, and lipid rich vesicles was achieved by observing CH2 stretching vibrations of lipids. The image acquisition rate of 5 frames per second was achieved without any staining. We also demonstrated real-time CARS imaging of laser-induced disruption and reaction of organelles in a living HeLa cell. A near-infrared pulsed laser beam tightly focused on an organelle in a living cell produces ablation at the focal point, causing local disruption of the organelle. We visualized the spatial and temporal distributions of a lipid rich organelles in the cytoplasm of a living HeLa cell in laser-induced dissection. We also demonstrated real-time CARS imaging of disruption of a plasma membrane and its repair. © 2010 Copyright SPIE - The International Society for Optical Engineering.
  • 新岡 宏彦, 蘆田幸一郎, 吉木 啓介, 荒 木 勉, 橋 本 守
    光学 39 8 401-403  2010年 [査読有り][招待有り]
  • Mamoru Hashimoto, Takeo Minamikawa, Tsutomu Araki
    MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES X 7569 75690Q  2010年 [査読有り][通常論文]
     
    We developed a high speed CARS (coherent anti-Stokes Raman scattering) spectral-imaging system using an acousto optic tunable filter and multi-focus excitation system. We compared two methods of CARS emission filtering and CARS excitation filtering. In both case, two laser pulses with narrow band (picosecond laser) and broad band (femotosecond laser) were used for the light source. For CARS emission filtering, the generated CARS was filtered by an AOTF, and for excitation filtering the broad band femtosecond laser pulse were filtered by an AOTF before excitation. The experimental results indicated that the CARS emission filtering was suitable for CARS microscopy.
  • Takeo Minamikawa, Mamoru Hashimoto, Katsumasa Fujita, Satoshi Kawata, Tsutomu Araki
    OPTICS EXPRESS 17 12 9526 - 9536 2009年06月 [査読有り][通常論文]
     
    We developed a multi-focus excitation coherent anti-Stokes Raman scattering (CARS) microscope using a microlens array scanner for real-time molecular imaging. Parallel exposure of a specimen with light from two highly controlled picosecond mode-locked lasers (jitter of 30 fs through an electronic low-pass filter with 150 Hz bandwidth, point-by-point wavelength scan within 300 ms) and parallel detection with an image sensor enabled real-time imaging. We demonstrated real-time CARS imaging of polystyrene beads (frame rate of 30 fps), a giant multi-lamellar vesicle of dipalmitoylphosphatidylcholine (frame rate of 10 fps), and living HeLa cells (frame rate of 10 fps). (C) 2009 Optical Society of America
  • Mamoru Hashimoto, Koichiro Ashida, Keisuke Yoshiki, Tsutomu Araki
    OPTICS LETTERS 34 9 1423 - 1425 2009年05月 [査読有り][通常論文]
     
    We have studied the enhancement of second-harmonic generation (SHG) from self-assembled monolayers on Au surfaces excited by radially polarized beams. The electric field at the metal surface was enhanced by constructive interference between the incident and the reflected beams due to a longitudinal field, which is the field parallel to the optical axis, generated around the focus by the radially polarized beam. Since even-order nonlinear phenomena are surface sensitive, the combination of SHG and a radially polarized beam has the potential to be a powerful new imaging tool for characterization of metal surfaces. The SHG signal excited by the radially polarized beam was about 3 times higher than that excited by a linearly polarized beam; in addition, the SHG from a 7-(dimethylamino)-4-methylcoumarin-3-isothiocyanate monolayer was about 1.3 times higher than that from a bare Au. substrate. (C) 2009 Optical Society of America
  • Mamoru Hashimoto, Takeo Minamikawa, Tsutomu Araki, Katsumasa Fujita, Satoshi Kawata
    Progress in Biomedical Optics and Imaging - Proceedings of SPIE 7183 2009年 [査読有り][通常論文]
     
    We realized realtime molecular imaging with low excitation laser intensity using a multi-focus excitation CARS (coherent anti-Stokes Raman scattering) microscope. We demonstrated realtime CARS images of polystyrene beads and lipid vesicles. Time series CARS images of the polystyrene beads in water was obtained with the frame rate of 30 fps. The three dimensional lipids vesicle which consists of 50 slices was observed within 7 s (100 ms/image). © 2009 SPIE.
  • Takeo Minamikawa, Tsutomu Araki, Mamoru Hashimoto
    Progress in Biomedical Optics and Imaging - Proceedings of SPIE 7183 2009年 [査読有り][通常論文]
     
    We demonstrated high-speed imaging of the distribution of DPPC (dipalmitoylphosphatidylcholine), d62-DPPC (deuterated DPPC), and DOPC (dioleoylphosphatidylcholine) lipids in a lipid vesicle with a multi-focus ex-citation CARS (coherent anti-Stokes Raman scattering) microscope using a microlens array scanner. By the multi-focus excitation, the dwell time is increased in proportion to the number of focal spots compared with a single beam scanning, and high-speed and high-quality CARS imaging is possible without increasing the peak power of each spot. We demonstrated the selectively visualization of DPPC and d62-DPPC lipid vesicles, in which the vesicles contain a type of lipid, by observing at 2840 cm-1 and 2090 cm-1. We also visualized the DOPC and DPPC lipids distribution in a lipid mixture vesicle observed at 1440 cm-1 and 1655 cm-1. The image acquisition time of 10 s/image at each Raman shift was realized. The signal ratio of 1440 cm-1 and 1655 cm-1 was locally intense on the lipid vesicle. It must be because the gel phase domain of DPPC lipids was exists in the DOPC lipids which were liquid-crystalline phase at room temperature. © 2009 SPIE.
  • Mamoru Hashimoto, Takeo Minamikawa, Hirohiko Niioka, Tsutomu Araki
    Proceedings of SPIE - The International Society for Optical Engineering 7507 75070H  2009年 [査読有り][通常論文]
     
    We developed a multifocus excitation coherent anti-Stokes Raman scattering microscope using a microlens array scanner for realtime molecular imaging. Two picoseond mode-locked lasers tightly synchronized were splited to a few tens of foci with the microlens array, the foci excited the sample parallely and the generated CARS from each spot was detected with an image sensor at once. By the multifocus excitation, exposure time was prolonged proportionally to the number of the foci because of parallel excitation and detection. The video-rate (frame rate of 30 fps) imaging of polystyrene beads in water was demonstrated, and the Brownian motion of beads were clearly obtained. The three-dimensional reconstructed imaging of living HeLa cells (frame rate of 5 fps, 85 images) was also demonstrated. © 2009 SPIE.
  • Mamoru Hashimoto, Takeo Minamikawa, Tsutomu Araki, Katsumasa Fujita, Satoshi Kawata
    MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES IX 7183 2009年 [査読無し][通常論文]
     
    We realized realtime molecular imaging with low excitation laser intensity using a multi-focus excitation CARS (coherent anti-Stokes Raman scattering) microscope. We demonstrated realtime CARS images of polystyrene beads and lipid vesicles. Time series CARS images of the polystyrene beads in water was obtained with the frame rate of 30 fps. The three dimensional lipids vesicle which consists of 50 slices was observed within 7 s (100 ms/image).
  • Takeo Minamikawa, Tsutomu Araki, Mamoru Hashimoto
    MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES IX 7183 718328  2009年 [査読有り][通常論文]
     
    We demonstrated high-speed imaging of the distribution of DPPC (dipalmitoylphosphatidycholine), d62-DPPC (deuterated DPPC), and DOPC (dioleoylphosphatidylcholine) lipids in a lipid vesicle with a multi-focus excitation CARS (coherent anti-Stokes Raman scattering) microscope using a microlens array scanner. By the multi-focus excitation, the dwell time is increased in proportion to the number of focal spots compared with a single beam scanning, and high-speed and high-quality CARS imaging is possible without increasing the peak power of each spot. We demonstrated the selectively visualization of DPPC and d62-DPPC lipid vesicles, in which the vesicles contain a type of lipid,, by observing at 2840 cm(-1) and 2090 cm(-1). We also visualized the DOPC and DPPC lipids distribution in a lipid mixture vesicle observed at 1440 cm(-1) and 1655 cm(-1). The image acquisition time of 10 s/image at each Raman shift was realized. The signal ratio of 1440 cm(-1) and 1655 cm(-1) was locally intense on the lipid vesicle. It must be because the gel phase domain of DPPC lipids was exists in the DOPC lipids which were liquid-crystalline phase at room temperature.
  • 吉木 啓介, 阿井川智正, 橋 本 守, 栗 原 誠, 橋本 信幸, 荒 木 勉
    生体医工学 46 6 698 - 702 2008年 [査読有り][通常論文]
  • Takeshi Yasui, Ken-ichi Sawanaka, Atsushi Ihara, Emmanuel Abraham, Mamoru Hashimoto, Tsutomu Araki
    OPTICS EXPRESS 16 2 1208 - 1221 2008年01月 [査読有り][通常論文]
     
    Terahertz time-domain spectroscopic (THz-TDS) imaging is an interesting new tool for nondestructive testing and other applications. However, the current speed of image acquisition is relatively low, making it difficult to use for moving objects. In this paper, we propose a real-time THz-TDS line scanner based on electro-optical time-to-space conversion and line focusing of a THz beam. The proposed system functions as a color scanner in the terahertz spectral region with fast line-scanning and has been successfully used to image objects, which are moved on a translation stage. The achieved THz-TDS imaging rate is 23 200 pixels per second. This proposed THz-TDS line scanner has the potential to become a powerful tool for monitoring moving objects in various real-world applications. (c) 2008 Optical Society of America.
  • Keisuke Yoshiki, Ryosuke Kanamaru, Mamoru Hashimoto, Nobuyuki Hashimoto, Tsutomu Araki
    OPTICS LETTERS 32 16 2465 - 2465 2007年08月 [査読有り][通常論文]
  • Keisuke Yoshiki, Kanamaru Ryosuke, Mamoru Hashimoto, Tsutomu Araki, Nobuyuki Hashimoto
    OPTICS LETTERS 32 12 1680 - 1682 2007年06月 [査読有り][通常論文]
     
    We developed a compact polarization-mode converter for microscopy to control three-dimensional polarization at the focus. The converter consisted of two homogeneously aligned liquid-crystal spatial light modulators with eight independently controllable electrodes (segments), and a quarter-waveplate. The converter converted a linearly polarized beam to three polarization modes: two orthogonal linear polarizations and a pseudo-radial polarization. We applied the converter to second-harmonic-generation microscopy and demonstrated the detection of three-dimensional molecular orientation. (C) 2007 Optical Society of America
  • Mamoru Hashimoto, Keisuke Yoshiki, Ryosuke Kanamaru, Nobuyuki Hashimoto, Tsutomu Araki
    THREE-DIMENSIONAL AND MULTIDIMENSIONAL MICROSCOPY: IMAGE ACQUISITION AND PROCESSING XIV 6443 64430L  2007年 [査読有り][通常論文]
     
    We developed a compact polarization-converter using two liquid-crystal spatial-light-modulators with eight electrodes. The converter converted a linearly polarized beam to two orthogonal linearly polarized beams and a radially polarized beam, and the direction of the electric filed at the focal point were controlled three-dimensionally. We constructed a second-harmonic-generation microscope using the polarization-converter to observe three-dimensional molecular orientation and demonstrated the detectability of molecular orientation.
  • Takeo Minamikawa, Naoki Tanimoto, Mamoru Hashimoto, Tsutomu Araki, Minoru Kobayashi, Katsumasa Fujita, Satoshi Kawata
    APPLIED PHYSICS LETTERS 89 19 191101  2006年11月 [査読有り][通常論文]
     
    The authors have developed a highly synchronized picosecond mode-locked laser system. A balanced cross-correlator using two-photon detectors was employed to observe femtosecond order timing jitter between two picosecond lasers (1.26 fs with 150 Hz bandwidth and 7.14 fs with 1 kHz bandwidth), and a signal from the correlator was used as a feedback control signal to reduce the timing jitter. The timing jitter between the two lasers was reduced to 8 fs through a low-pass filter with 150 Hz bandwidth.
  • K. Yoshiki, M. Hashimoto, T. Araki
    LASER BEAM SHAPING VII 6290 62900F  2006年 [査読有り][通常論文]
     
    We have developed a second-harmonic-generation (SHG) microscope to observe the three-dimensional molecular orientation with three-dimensional high spatial resolution using a polarization mode converter. The mode converter consists of a parallel-aligned nematic-liquid-crystal spatial-light-modulator (PAL-SLM) and quarter-waveplates, and converts a incident linearly polarized beam to orthogonal linearly polarized beams or radially polarized beam. We combined the mode converter with SHG microscope to obtain the local information of the three-dimensional molecular orientation. We demonstrated the detection of three-dimensional molecular orientation of collagen fiber in human Achilles' tendon. For high precision three-dimensional molecular orientation measurement, we propose a technique to calibrate the dependence of SHG detection efficiencies on molecular orientation using a liposome.
  • K Yoshiki, H Azuma, K Yoshioka, M Hashimoto, T Araki
    OPTICAL REVIEW 12 5 415 - 419 2005年09月 [査読有り][通常論文]
     
    We have investigated the fluorescence lifetime properties of 8 calcium ion probes, calcium-green-1, calcium green-2, calcium green-5N, calcium orange, oregon green 488 BAPTA-6F, fluo-3, fluo-4, and fluo-5N. We found that the decay time of calcium green-5N varied more sensitively with calcium concentration than calcium green-1 which was known to be a highly sensitive probe. We have also found that the center of observable range of calcium concentration by fluorescence lifetime measurement is lower than that by fluorescence intensity measurement. (c) 2005 The Optical Society of Japan.
  • K. Yoshiki, M. Hashimoto, T. Araki
    2005 Pacific Rim Conference on Lasers & Electro-Optics 2005年07月 [査読有り][通常論文]
  • M Hashimoto, T Asada, T Araki, Y Inouye, S Kawata
    JAPANESE JOURNAL OF APPLIED PHYSICS PART 1-REGULAR PAPERS BRIEF COMMUNICATIONS & REVIEW PAPERS 44 6A 3958 - 3961 2005年06月 [査読有り][通常論文]
     
    We have proposed and developed a new feedback control system to automatically minimize the pulse duration of an ultrafast laser using a two-photon absorption detector that has a Gires-Toumois interferometer (GTI) for compensating the positive group delay dispersion of the laser cavity. The signal of the two-photon detector is inversely proportional to the pulse duration when the average power, repetition rate, and pulse shape are maintained. The minimization of the pulse duration is accomplished by adjusting the voltage applied to the GTI to maximize the two-photon signal. We demonstrated the control of pulse duration minimization with the developed system.
  • M Hashimoto, T Asada, T Araki, S Kawata
    2005 PACIFIC RIM CONFERENCE ON LASERS AND ELECTRO-OPTICS 660 - 661 2005年 [査読有り][通常論文]
     
    We have developed the automatic pulse duration control system using two-photon absorption detector for stable operation and pulse duration control. We have applied the developed system to the multi-focus CARS microscopy.
  • S Kawata, T Ichimura, N Hayazawa, M Hashimoto, Y Inouye
    MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES V 5700 52 - 59 2005年 [査読有り][通常論文]
     
    Optical microscopy that can visualize the molecular vibration with a nanometric spatial resolution has been realized by a combination of near-field optics and coherent anti-Stokes Raman scattering (CARS) spectroscopy. A metallic probe with a sharp tip is used to strongly enhance optical near-field in the local vicinity of the tip owing to the excitation of local surface plasmon polariton. CARS signals of molecules in the local area can be strongly induced by the plasmonic field. We have visualized DNA molecules and single-walled carbon nanotubes (SWNTs) with a spatial resolution far beyond the diffraction limit by the tip-enhanced near-field CARS microscopy.
  • K Yoshiki, M Hashimoto, T Araki
    JAPANESE JOURNAL OF APPLIED PHYSICS PART 2-LETTERS & EXPRESS LETTERS 44 33-36 L1066 - L1068 2005年 [査読有り][通常論文]
     
    We have developed a second-harmonic-generation (SHG) microscope using an excitation beam with a controlled polarization pattern in order to detect three-dimensional molecular orientation. The electric field at the focus is controlled three-dimensionally by modifying the polarization distribution with a parallel-aligned nematic-liquid-crystal spatial-light-modulator without any mechanical moving parts. We demonstrated that the SHG signal from an Achilles tendon, sliced so that collagen fibers were aligned parallel to the optical axis, excited by a radially polarized beam was higher than those excited by linearly polarized beams. The possibility of determinating three-dimensional molecular orientation was thus shown.
  • M Hashimoto, K Yamada, T Araki
    OPTICAL REVIEW 12 1 37 - 41 2005年01月 [査読有り][通常論文]
     
    We have proposed a method to control the three-dimensional electric field in the focus of an optical microscope using two non-twisted liquid crystal spatial light modulators, and to detect the molecular orientation of a single molecule. The three-dimensional electric field is generated by focusing the beam with two dimensional spatial distribution of polarization. The possibility of detection of three-dimensional single molecular orientation was shown by numerical calculations. (c) 2005 The Optical Society of Japan.
  • S Kawata, T Ichimura, N Hayazawa, Y Inouye, M Hashimoto
    JOURNAL OF NONLINEAR OPTICAL PHYSICS & MATERIALS 13 3-4 593 - 599 2004年12月 [査読無し][招待有り]
     
    We apply the field enhancement effect due to plasmon polariton excitation on a metallic nanostructure in order to improve the diffraction limited spatial resolution of coherent anti-Stokes Raman scattering (CARS) microscopy. A cantilever probe tip coated with a 25 nm-thick gold film is utilized as a near-field light source to locally excite the CARS polarizations near the tip. Our CARS microscope has effectively enhanced the CARS signals and realized vibrational imaging of single-wall carbon nanotubes (SWNTs) beyond the spatial resolution of far-field CARS microscopy.
  • T Ichimura, N Hayazawa, M Hashimoto, Y Inouye, S Kawata
    PHYSICAL REVIEW LETTERS 92 22 220801  2004年06月 [査読有り][通常論文]
     
    An electric field enhanced by a metallic nanoprobe has locally induced coherent anti-Stokes Raman scattering (CARS) of adenine molecules in a nanometric DNA network structure. Owing to the third-order nonlinearity, the excitation of the CARS polarization is extremely confined to the end of the tip apex, resulting in a spatial resolution far beyond the diffraction limit of light. Our tip-enhanced CARS microscope visualized the DNA network structure at a specific vibrational frequency (similar to1337 cm(-1)) corresponding to the ring-breathing mode of diazole of adenine molecules.
  • N Hayazawa, T Ichimura, M Hashimoto, Y Inouye, S Kawata
    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 227 U280 - U280 2004年03月 [査読有り][通常論文]
  • N Hayazawa, T Ichimura, M Hashimoto, Y Inouye, S Kawata
    JOURNAL OF APPLIED PHYSICS 95 5 2676 - 2681 2004年03月 [査読有り][通常論文]
     
    On the basis of the mechanism of surface enhanced Raman scattering, it is shown that coherent anti-Stokes Raman scattering (CARS) of molecules attached to isolated gold nanoparticles are strongly enhanced and the signal from each particle is well localized. In addition to well-known advantages of CARS, the surface enhanced CARS combined with a scanning system of metallic nanoprobe tip can realize high spatial resolution CARS microscopy beyond the diffraction limit of light by locally enhancing the weak signals from the small sample volume. This concept is realized by tip-enhanced coherent anti-Stokes Raman spectroscopy using a metallic nanoprobe of near-field scanning optical microscope. (C) 2004 American Institute of Physics.
  • T Ichimura, N Hayazawa, M Hashimoto, Y Inouye, S Kawata
    APPLIED PHYSICS LETTERS 84 10 1768 - 1770 2004年03月 [査読有り][通常論文]
     
    A tip-enhanced electric field at a metallic probe tip of apertureless near-field scanning optical microscope was applied to a third-order nonlinear optical process, coherent anti-Stokes Raman spectroscopy. The combination of the enhanced field and third-order nonlinearity resolved molecular vibrations of adenine molecules embedded in deoxyribonucleic acid double-helix nanocrystals beyond the diffraction limit of light. (C) 2004 American Institute of Physics.
  • S Kawata, T Ichimura, N Hayazawa, M Hashimoto, Y Inouye
    NONLINEAR OPTICAL TRANSMISSION AND MULTIPHOTON PROCESSES IN ORGANICS II 5516 1 - 8 2004年 [査読有り][通常論文]
     
    A metallic nano-probe has locally induced coherent anti-Stokes Raman scattering (CARS) of adenine molecules in a nanometric DNA network structure. The excitation fields and CARS polarization are enhanced by the tip apex of the nano-probe through the excitation of local surface plasmons. Owing to the third-order nonlinearity- the excitation of the CARS polarization is extremely confined to the end of the tip apex. resulting in the spatial resolution far beyond the diffraction limit of light. Our CARS microscope using a silver-coated probe visualized the DNA network structure at a specific vibrational frequency (similar to1337 cm(-1)) of adenine molecules with a spatial resolution of similar to15 nm and sufficient sensitivity.
  • S Kawata, N Hayaazawa, T Yano, H Watanabe, T Ichimura, M Hashimoto, Y Inouye
    NANOBIOPHOTONICS AND BIOMEDICAL APPLICATIONS 5331 1 - 12 2004年 [査読有り][通常論文]
     
    A light microscope capable to show images of molecules in nanometer scale has been a dream of scientists, which, however, is difficult clue to the strict limitation of spatial resolution due to the wave nature of light [1]. While there have been attempts to overcome the diffraction limit by using nonlinear response of materials [2, 3], near-field optical microscopy could provide better detecting accuracy [4-6]. In this paper, we present molecular distribution nano-imaging colored by Raman-scattering spectral shifting, which is probed with a metallic tip. The metallic probe tip has been used to enhance the optical field only in the vicinity of probe tip [7-11]. The effect is similar to the one seen in the detection of molecules on the metal-island film, known as surface-enhanced Raman spectroscopy (SERS) [12], while in this case a single metallic tip works for the field enhancement in nanometer scale.
  • T Ichimura, N Hayazawa, M Hashimoto, Y Inouye, S Kawata
    JOURNAL OF RAMAN SPECTROSCOPY 34 9 651 - 654 2003年09月 [査読有り][通常論文]
     
    We observed local enhancement of coherent anti-Stokes Raman scattering (CARS) by isolated gold nanoparticles. CARS signals from adenine molecules of a DNA base attached to isolated gold nanoparticles were strongly enhanced owing to the electromagnetic enhancement at the local proximity of the particles. The localization of the enhancement was successfully observed by a CARS microscope using a collinear configuration of tightly focused lasers. The CARS spectrum of adenine was enhanced by a factor of 2000 and was in good agreement with the measured spontaneous Raman spectrum. Copyright (C) 2003 John Wiley Sons, Ltd.
  • 伊丹 伸, 橋本 守, 荒木 勉
    日本機会学会論文集(C編) 69 684 210 - 215 2003年 [査読有り][通常論文]
  • M Hashimoto
    ACTA HISTOCHEMICA ET CYTOCHEMICA 35 2 83 - 86 2002年 [査読有り][招待有り]
     
    In this paper, new multiphoton microscopy using CARS (coherent anti-Stokes Raman scattering) spectroscopy is described. CARS microscopy has the features of non-staining molecular mapping by molecular vibration imaging and three-dimensional resolution by multiphoton process. A picosecond tunable laser and suitable optical filters provide the CARS imaging in the fingerprint region, and multi-focus excitation using a rotatory-microlens array enables the multi-spectral imaging.
  • Y Tohno, S Tohno, Y Tateyama, Y Kida, T Yasui, M Hashimoto, T Araki
    BIOLOGICAL TRACE ELEMENT RESEARCH 81 2 115 - 125 2001年08月 [査読無し][招待有り]
     
    To elucidate the calcium content of the arteries in the upper and lower limbs, the authors determined the calcium content of all the arteries in the upper and lower limbs continuously by microwave-induced plasma-atomic emission spectrometry. The subjects were an 87-yr-old man and a 72-yr-old woman. The calcium content was determined both in the arteries of the upper limbs continuously, such as the subclavian arteries and its distal arteries, and in the arteries of the lower limbs, such as the common iliac arteries and its distal arteries. The common finding that the higher accumulation of calcium occurred in the arteries of the lower limbs in comparison to the arteries of the upper limbs and extremely high accumulation of calcium occurred in the common, external, and internal iliac arteries was obtained in the two subjects. The calcium content of the arteries in the upper and lower limbs was visually demonstrated.
  • M Hashimoto, T Araki
    JOURNAL OF THE OPTICAL SOCIETY OF AMERICA A-OPTICS IMAGE SCIENCE AND VISION 18 4 771 - 776 2001年04月 [査読有り][通常論文]
     
    The three-dimensional coherent transfer function of confocal coherent anti-Stokes Raman scattering microscopy was derived theoretically. The three-dimensional optical transfer function was also derived under the weak-contrast assumption. The effect of a pinhole in front of the detector on the optical transfer function was estimated, and it was found that the cutoff frequency of the optical transfer function is independent of the pinhole. Micrometer-order spatial resolution along the optical axis was also experimentally demonstrated. (C) 2001 Optical Society of America.
  • Mamoru Hashimoto, Tsutomu Araki
    Journal of the Optical Society of America A: Optics and Image Science, and Vision 18 4 771 - 776 2001年 [査読有り][通常論文]
     
    The three-dimensional coherent transfer function of confocal coherent anti-Stokes Raman scattering microscopy was derived theoretically. The three-dimensional optical transfer function was also derived under the weak-contrast assumption. The effect of a pinhole in front of the detector on the optical transfer function was estimated, and it was found that the cutoff frequency of the optical transfer function is independent of the pinhole. Micrometer-order spatial resolution along the optical axis was also experimentally demonstrated. © 2001 Optical Society of America.
  • M Hashimoto, T Araki, S Kawata
    OPTICS LETTERS 25 24 1768 - 1770 2000年12月 [査読有り][通常論文]
     
    We have developed a new coherent anti-Stokes Raman scattering (CARS) microscopy system with a collinear configuration for use in the fingerprint region. The system consists of a picosecond laser system and a transmission-type laser scanning microscope without a pinhole in front of the detector. The observable Raman-shift region is 900-1750 cm(-1), the spectral resolution is 30 cm(-1), and the spatial resolution is smaller than 1 mum in the lateral direction and 3.2 mum in the depth direction, with objectives with a numerical aperture of 0.65. CARS spectra and images of polystyrene beads are demonstrated, and CARS imaging of a viable yeast cell is attempted. (C) 2000 Optical Society of America.
  • Y Tateyama, Y Takano, Y Tohno, Y Moriwake, S Tohno, M Hashimoto, T Araki
    BIOLOGICAL TRACE ELEMENT RESEARCH 74 3 211 - 221 2000年06月 [査読有り][通常論文]
     
    To show the relationships of calcium accumulation in the thoracic aorta to the other tissues, calcium contents were determined with a microwave-induced plasma-atomic emission spectrometer on arteries, veins, cartilages, Ligaments, and bones. These tissues were resected from 18 individuals, consisting of 11 men and 7 women who died in the age range 59-91 yr. As thoracic and abdominal aortas are routinely used for radiographic examination of arterial calcification, they appear to be standard tissues of the calcium accumulation. The calcium accumulations were determined in the femoral artery, the superior and inferior venae cavae, the internal jugular vein, cartilages of the articular disk of the temporomandibular joint and the intervertebral disk, both the Ligaments of the anterior cruciate ligament and the ligamentum capitis femoris, and the calcaneus, in contrast with the thoracic aorta. As calcium increased in the thoracic aorta, it increased in the femoral artery, the articular disk of the temporomandibular joint, the intervertebral disk, both ligaments of the anterior cruciate ligament, and the Ligamentum capitis femoris, but it did not increase in veins, such as the superior and inferior venae cavae and the internal jugular vein. In contrast, it decreased in the calcaneus.
  • M Hashimoto, T Araki
    18TH CONGRESS OF THE INTERNATIONAL COMMISSION FOR OPTICS: OPTICS FOR THE NEXT MILLENNIUM, TECHNICAL DIGEST 3749 496 - 497 1999年 [査読有り][通常論文]
     
    We propose a new laser scanning microscope using coherent anti-Stokes Raman spectroscopy. As the proposed method is a kind of Raman spectroscopy, molecular structural informations are obtained without any staining. The imaging property is theoretically estimated by using the three-dimensional optical transfer function. It is shown that the proposed microscope has three-dimensional resolution in any case of measuring for the weak or the high contrast object with or without a pinhole before a detector. Spatial resolution of micrometer order along the optical axis is demonstrated. key words: Microscope, Nonlinear Optics, Raman spectroscopy.
  • Mamoru Hashimoto, Hiro-O. Hamaguchi
    Applied Spectroscopy 52 2 222 - 225 1998年 [査読有り][通常論文]
     
    The surface (about 130 molecular layers) of an oriented thin crystal of decanoic acid was subjected to sudden melting by a laser-induced temperature jump (T-jump), and the process of subsequent crystal re-growth was monitored by millisecond time-resolved multichannel Fourier transform infrared spectroscopy. The gauche-trans structural change of the alkane part of the molecule has been probed by the CH stretch bands in the 2800-3000 cm-1 region. The change in the molecular orientation has been detected by the OH stretch band around 3065 cm-1. The recovery curves for the CH2 antisymmetric stretch and the OH stretch bands are markedly different from each other in the first 200 ms, suggesting that the gauche-trans structural changes precedes the crystal re-growth. After 500 ms, the recovery curves become identical. This result means that the rate of the gauche to the trans structural change is equal to the rate of the recovery of the molecular orientation. It is highly likely that a fast equilibrium is attained between the gauche and the trans conformations in the liquid phase after 500 ms from the sudden melting and that the crystal re-growth takes place solely via the all-trans structure in the liquid phase.
  • K Tokumura, M Natsume, T Nakagawa, M Hashimoto, T Yuzawa, H Hamaguchi, M Itoh
    CHEMICAL PHYSICS LETTERS 271 4-6 320 - 326 1997年06月 [査読有り][通常論文]
     
    Time-resolved infrared absorption spectra of 7-hydroxyquinoline (7-HQ) in methanol were measured to investigate the relaxation processes following S-l --> S-l proton transfer tautumerization. Deuterium and nitrogen isotope effects were observed for the transient infrared spectra of 7-HQ-N-14 and -N-15 in MeOD and MeOH. The 1644 (1628) cm(-1) band in MeOH (MeOD) is ascribed to the H(D)-bonded C = O stretching of the phototautomer in the ground state (S-0'). Transient absorption decay exhibits a remarkable deuterium isotope effect, It is thus demonstrated that the ground-state reverse proton transfer of S-0', is responsible for the observed transient decay.
  • 藤澤 泰充, 橋本 守, 荒木 勉
    照明学会誌 81 8A 656 - 663 1997年05月 [査読有り][通常論文]
  • Tsutomu Araki, Yasumitsu Fujisawa, Mamoru Hashimoto
    Review of Scientific Instruments 68 3 1365 - 1368 1997年 [査読有り][通常論文]
     
    An optical function pulse generator that emits (1) short pulse of 1 ns duration, (2) double pulse with variable time interval, and (3) square waveform pulse of variable width in nanosecond range is devised using an InGaN/AlGaN double heterostructure light emitting diode (LED). Although the LED emits a 450 nm (blue) light under conventional dc operation below 30 mA, 380 nm light due to the InGaN/AlGaN component appears when a current larger than 50 mA is applied. This phenomenon is used to realize a pulsed ultraviolet light source. Under large nanosecond current pulsing (peak current > 1 A), an intense pulsed emission of 380 nm is obtained. Pulse waveform of the LED emission can be adjusted electrically by applying a shaped current to the LED. To evaluate the potential of the pulse generator as a test source of photodetectors, the response waveforms of photomultiplier tubes were measured. © 1997 American Institute of Physics.
  • M Hashimoto, HO Hamaguchi
    APPLIED SPECTROSCOPY 50 8 1030 - 1033 1996年08月 [査読有り][通常論文]
     
    A newly designed multichannel Fourier transform infrared spectrometer has been constructed for single-event time-resolved spectroscopy. It is capable of measuring unrepeatable transient events such as phase transitions, explosions, etc., with a time resolution of up to 5.14 ms. The mid-infrared region of 4500-2500 cm(-1) is covered with a maximum spectral resolution of 13 cm(-1). The developed system is used to measure the phase transitions of alkanes. The solid-solid and solid-liquid phase transitions of octadecane and nonadecane have been studied with time resolutions of 5-50 ms.
  • Mamoru Hashimoto, Hiro-o Hamaguchi
    The Journal of Physical Chemistry 99 20 7875 - 7877 1995年05月 [査読有り][通常論文]
  • 橋本 守, 河田 聡
    分光研究 41 5 317 - 326 1992年08月 [査読有り][通常論文]
     
    The signal-to-noise ratio of multi-channel Fourier-transform spectroscopy is compared with those of multi-channel dispersive spectroscopy, conventional Fourier-transform spectroscopy and conventional dispersive spectroscopy in the detector-noise or the photon-noise dominant cases. The results show that multi-channel Fourier-transform spectroscopy is not superior to others with the same optical thorouput but higher signal-to-noise ratio is obtained in the case when its large optical throughput can be utilized.
  • Mamoru Hashimoto, Satoshi Kawata
    Applied Optics 31 28 6096 - 6101 1992年 [査読有り][通常論文]
     
    A compact Fourier-transform IR spectrometer without a moving mechanism was developed. The spectrometer consists of a shearing interferometer for forming a spatially distributed interferogram and an IR array detector for observing the interferogram. The shearing interferometer of the developed system is a birefringent interferometer with a Savert plate the IR array detector is a PtSi Schottky- barrier detector with 4096 elements. The optics and the system configuration are described in detail, and the experimental results of the IR absorption spectra of polystyrene and polyethylene terephthalate film are shown. The developed optics is as small as 20 x 6 cm-1 in size. The spectral resolution of the prototype system is ∼27.6 cm-1 between 5000 and 2000 cm-1. The methods and their possibilities of resolution improvement are also described. © 1992 Optical Society of America.
  • Satoshi Kawata, Mamoru Hashimoto
    ANALYTICAL SCIENCES 7 575 - 576 1991年 [査読有り][通常論文]
     
    A multichannel Fourier-transform spectrometer in infrared was developed with birefringent interferometer with a Saverts plate and a infrared CCD with 4096 elements. The optics and the system configuration are described, and some experimental results of infrared spectrum measurement are shown for standard samples and an infrared radiation source. The resolution attained by the present system is -27.6 cm(-1) between 5000-2000 cm(-1)

書籍

  • Raman Imaging
    M. Hashimoto, T. Ichimura, K. Fujita (担当:分担執筆範囲:CARS microscopy: Implementation of nonlinear vibrational spectroscopy for far-field and near-field imaging)
    Springer 2012年 168
  • 顕微分光法 ナノ・マイクロの世界を見る分光法
    (担当:分担執筆範囲:赤外・ラマン顕微分光法)
    講談社サイエンティフィク 2009年
  • バイオイメージングがわかる
    橋本守, 福島修一郎, 荒木勉 (担当:分担執筆範囲:LIM蛍光寿命イメージング)
    2005年
  • Biological Imaging and Sensing
    S. Kawata, O. Nakamura, T. Kaneko, M. Hashimoto, K. Goto, N. I. Smith, T. Sugiura, I. Fujimasa, H. Matsumoto (担当:分担執筆範囲:Biological imaging and sensing from basic techniques to clinical application)
    Springer 2004年 1-68
  • Mamoru Hashimoto, Teturo Yuzawa, Chihiro Kato, Koichi Iwata, Hiro‐o Hamaguchi (担当:分担執筆範囲:Fast time-resolved mid-infrared spectroscopy using grating spectrometers)
    John Wiley & Sons, Ltd 2001年 666-676

講演・口頭発表等

  • Label free bioimaging using nonlinear coherent Raman microscopy  [招待講演]
    M. Hashimoto
    2015 IEEE CPMT Symposium Japan (ICSJ) 2015年11月 口頭発表(招待・特別)
  • Nonlinear Raman microscopy for biomedical applications  [招待講演]
    M. Hashimoto, S. Kawahito
    2015年11月 口頭発表(一般)
  • Plasmonic-nanoparticle-enhanced hyper-Raman spectroscopy  [通常講演]
    B. Ranjan, M. Hashimoto, Y. Saito, P. Verma
    2015年09月 口頭発表(一般)
  • Label free imaging of atherosclerotic lesions using stimulate Raman scattering, second harmonic generation, and two-photon fluorescence microscopy  [通常講演]
    T. Aoki, T. Tao, S. Fukushima, T. Araki, M. Hashimoto
    Abstract of the 8th Asian-Pacific Conference on Biomechanics 2015年09月 口頭発表(一般)
  • Discrete fourier transform infrared spectroscopy using precisely periodic pulse  [通常講演]
    Y.-D. Hsieh, S. Okubo, H. Inaba, M. Hashimoto, T. Yasui
    2015年05月 口頭発表(一般)
  • マルチモーダル非線形光学顕微鏡によるアテローム性動脈 硬化症病変のイメージング”  [招待講演]
    橋本 守
    2015年05月 口頭発表(一般)
  • Biological high-resolution imaging in wet-condition using cathodoluminescence microscopy  [通常講演]
    T. Furukawa, S. Fukushima, H. Niioka, M. Ichimiya, J. Miyake, M. Ashida, T. Araki, M. Hashimoto
    Proceedings of The 5th Asian and Pacific-Rim Symposium on Biophotonics 2015年05月 口頭発表(一般)
  • 近赤外発光・カソードルミネッセンスによるマルチスケール生体観察  [通常講演]
    福島昌一郎, 新岡宏彦, 一宮正義, 三宅 淳, 芦田昌明, 荒木 勉, 橋本 守
    2015年03月
  • Highly luminescent rare-earth doped Y2O3 ceramics in near-infrared region by codoping Li+ ions and its application for imaging  [通常講演]
    J. Yamasaki, S. Fukushima, H. Niioka, T. Araki, M. Hashimoto, J. Miyake
    2014年12月
  • Gadolinium oxide doped rare earth nanophosphors for trimodal imaging  [通常講演]
    D. T, K. Dung, S. Fukushima, H. Niioka, M. Ichimiya, M. Ashida, T. Araki, M. Hashimoto, J. Miyake
    2014年12月
  • Observation of organic nonlinear optical crystal by multiplex fourth order Raman microscope  [通常講演]
    C. Ninagawa, T. furukawa, H. Niioka, T. Araki, M. Hashimoto
    2014年12月
  • 近赤外光・電子顕微鏡による相関バイオイメージング  [通常講演]
    福島昌一郎, 新岡宏彦, 一宮正義, 三宅 淳, 芦田昌明, 荒木 勉, 橋本 守
    2014年10月
  • 近赤外・カソードルミネッセンス相関観察を目指したナノ蛍光体粒子の開発  [通常講演]
    福島昌一郎, 新岡宏彦, 一宮正義, 三宅 淳, 芦田昌明, 荒木 勉, 橋本 守
    2014年07月
  • 希土類ナノ蛍光体を用いたマルチカラーカソードルミネッセンス・アップコンバージョン生体観察  [通常講演]
    福島昌一郎, 古川太一, 新岡宏彦, 一宮正義, 芦田昌明, 三宅 淳, 荒木 勉, 橋本 守
    2014年03月
  • 電子線及び近赤外光照射による発光を用いたバイモーダル生体観察  [通常講演]
    福島昌一郎, 古川太一, 新岡宏彦, 一宮正義, 芦田昌明, 三宅 淳, 荒木 勉, 橋本 守
    2013年11月
  • マルチスケール生体イメージングを目指したカソードルミネッセンス・アップコンバージョンナノ蛍光体の作製  [通常講演]
    福島昌一郎, 古川太一, 新岡宏彦, 一宮正義, 芦田昌明, 三宅 淳, 荒木 勉, 橋本 守
    2013年11月
  • マルチスケール相関生体イメージングを実現するバイモーダルナノ蛍光体の合成  [通常講演]
    福島昌一郎, 古川太一, 新岡宏彦, 一宮正義, 三宅 淳, 芦田昌明, 荒木 勉, 橋本 守
    2013年10月
  • アップコンバージョンナノ蛍光体を用いたCL・蛍光イメージング  [通常講演]
    H. Niioka, T. Furukawa, S. Fukushima, M. Ichimiya, T. Nagata, J. Miyake, M. Ashida, T. Araki, M. Hashimoto
    2013年10月
  • カソードルミネッセンス顕微鏡を用いたバイオイメージング  [通常講演]
    古川太一, 新岡宏彦, 福島昌一郎, 一宮正義, 市川 聡, 永田智啓, 芦田昌明, 荒木 勉, 橋本 守
    2013年07月
  • アップコンバージョン発光とカソードルミネッセンスによる生体観察を目指した希土類ナノ蛍光体プローブの作製  [通常講演]
    福島昌一郎, 古川太一, 新岡宏彦, 一宮正義, 永田智啓, 三宅 淳, 芦田昌明, 荒木 勉, 橋本 守
    2013年03月
  • カソードルミネッセンス生体イメージングのための微小希土類添加ナノ蛍光体作製  [通常講演]
    古川太一, 新岡宏彦, 一宮正義, 市川 聡, 永田智啓, 三宅 淳, 芦田昌明, 荒木 勉, 橋本 守
    2013年03月
  • 希土類添加Y2O3ナノ蛍光体を用いたマルチモーダル蛍光・CL細胞イメージング  [通常講演]
    新岡宏彦, 古川太一, 一宮正義, 永田智啓, 芦田昌明, 荒木 勉, 橋本 守
    2012年11月
  • 光学顕微鏡とカソードルミネッセンス顕微鏡を用いたマルチモーダル細胞イメージング  [通常講演]
    新岡宏彦, 古川太一, 一宮正義, 永田智啓, 芦田昌明, 荒木勉, 橋本守
    2012年09月
  • ナノ蛍光体粒子とカソードルミネッセンス顕微鏡を用いたマルチカラー生体イメージング  [招待講演]
    新岡宏彦, 古川太一, 一宮正義, 芦田昌明, 荒木勉, 橋本守
    日本顕微鏡学会 (つくば国際会議場, 2012/5/14-16, 招待講演) 2012年05月
  • 希土類ナノ蛍光体を用いた生体カソードルミネッセンスイメージングの高輝度化  [通常講演]
    古川太一, 新岡宏彦, 一宮正義, 永田智啓, 芦田昌明, 荒木 勉, 橋本 守
    2012年03月
  • ナノ蛍光体粒子を用いたカソードルミネッセンス生体イメージング  [通常講演]
    新岡宏彦, 古川太一, 一宮正義, 芦田昌明, 荒木 勉, 橋本 守
    2012年02月
  • ナノ蛍光体粒子を用いたマルチカラー生体カソードルミネッセンスイメージング  [通常講演]
    古川太一, 新岡宏彦, 一宮正義, 永田智啓, 芦田昌明, 荒木 勉, 橋本 守
    2011年11月
  • カソードルミネッセンスを用いた細胞の超解像イメージング  [通常講演]
    古川太一, 新岡宏彦, 一宮正義, 芦田昌明, 荒木 勉, 橋本 守
    2011年09月
  • カソードルミネッセンスを利用した生体細胞の超解像イメージング手法  [通常講演]
    古川太一, 新岡宏彦, 一宮正義, 芦田昌明, 荒木 勉, 橋本 守
    2011年07月
  • Realtime biomolecular imaging by multifocus CARS microscopy  [通常講演]
    2009年
  • High-speed non-staining biomolecular imaging by nonlinear Raman microspectroscopy  [通常講演]
    2009年
  • Multi-focus CARS microscopy using automatic pulse duration control system  [通常講演]
    Proceedings of International Conference on Quantum Electronics 2005 and the Pacific Rim Conference on Lasers and Electro-Optics 2005 (IQEC/CLEO-PR 2005) 2005年
  • Coherent anti-Stokes Raman microscope for identification of cellular molecule  [通常講演]
    Proceedings of the Second Asian and Pacific Rim Symposium on Biophotonics 234-235 2004年
  • Fluorescence lifetime properties of various calcium ion indicators  [通常講演]
    Proc. ICO-14 2004年
  • Multi-spectral imaging by multi-focus CARS microscopy  [通常講演]
    Proceedings of Multi-dimensional Microscopy 2001, 23 2001年
  • Effect of the detector side pinhole on 3D optical properties of CARS microscopy  [通常講演]
    Proceedings of Focus on Microscopy 2000 2000年 口頭発表(一般)
  • Molecular Vibrational Imaging in the Fingerprint Region by Cars Microscopy  [通常講演]
    Proceedings of Focus on Microscopy 2000, p.~4 2000年

その他活動・業績

  • Keigo Hirose, Shuichiro Fukushima, Taichi Furukawa, Mamoru Hashimoto Optics InfoBase Conference Papers Part F78-JSAP 2017 paper 5p_A409_8 2017年 [査読有り][通常論文]
  • カソードルミネッセンス顕微鏡と近赤外顕微鏡によるマルチモーダルイメージング
    新岡宏彦, 福島昌一郎, 古川太一, 橋本守 光アライアンス 28 (1) 24 -28 2017年01月 [査読無し][招待有り]
  • Mamoru Hashimoto, Shoji Kawahito Transactions of Japanese Society for Medical and Biological Engineering 52 54 -SY-55 2014年08月17日 [査読有り][通常論文]
     
    We have developed a nonlinear optical multimodal microscopy system for diagnosis of atherosclerosis. By using a fast tuning picosecond mode-locked laser with an acoutso-optic tunable filter and modifying to synchronize with another mode-locked laser, fast spectral nonlinear Raman imaging becomes available. The combination of nonlinear Raman and second harmonic generation visualize lipid and collagen without any pre-treatment of sample. The developed system is applied to observe the artery of atherosclerotic lesion.
  • Tomoyo Tao, Harsono Cahyadi, Shuichiro Fukushima, Mamoru Hashimoto, Tsutomu Araki Transactions of Japanese Society for Medical and Biological Engineering 52 128 -O-129 2014年08月17日 [査読有り][通常論文]
     
    We have developed a nonlinear optical multimodal microscopy system for diagnostic tool of atherosclerosis. Nonlinear coherent optical imaging never requires invasive dyes to visualize the sample with three-dimensional resolution. In particular, coherent Raman scattering (CRS) and second harmonic generation (SHG) techniques provide label-free detection of lipid and collagen, respectively, which are keys of atherosclerotic plaques formation. Therefore, CRS/SHG multimodal imaging has a potential of quantitative diagnosis of atherosclerosis. In this study, we provide a histological assessment of abnormally produced collagen in aorta samples of mice, and show that the abnormally produced collagen has a correlation with the progression of lesion. Furthermore, we perform CRS/SHG imaging of the sample with the developed system, and lipid distribution and collagen in adventitia was visualized invasively.
  • 多焦点リアルタイムCARS顕微鏡による細胞応答観測
    橋本 守, Harsono Cahyadi, 南川 丈夫, 新岡 宏彦, 荒木 勉 オプトロニクス 33 (389) 74 -78 2014年05月 [査読無し][招待有り]
  • 希土類添加ナノ蛍光体粒子を用いたカソードルミネッセンス・蛍光細胞イメージング
    新岡 宏彦, 古川 太一, 橋本 守 顕微鏡 49 (1) 59 -63 2014年04月 [査読無し][招待有り]
  • Synthesis of nanophosphors for correlative cathodoluminescence, up-conversion, and near-infrared bioimaging
    S. Fukushima, T. Furukawa, H. Niioka, M. Ichimiya, J. Miyake, M. Ashida, T. Araki, M. Hashimoto 2014年03月 [査読無し][通常論文]
  • 高速広帯域波長走査レーザを光源とした多焦点CARS顕微鏡
    橋本 守, Harsono Cahyadi, 新岡 宏彦, 荒木 勉 光アライアンス 25 (3) 16 -20 2014年03月 [査読無し][招待有り]
  • Visible to near-infrared luminescent nanoparticles for multimodal bioimaging on nanometer to millimeter scale
    S. Fukushima, T. Furukawa, H. Niioka, M. Ichimiya, J. Miyake, M. Ashida, T. Araki, M. Hashimoto 2014年02月 [査読無し][通常論文]
  • 新岡 宏彦, 福島 昌一郎, 橋本 守, 荒木 勉, 小野島 大介, 湯川 博, 馬場 嘉信, 三宅 淳 日本生物工学会大会講演要旨集 66 256 -256 2014年 [査読無し][通常論文]
  • Shoichiro Fukushima, Hirohiko Niioka, Masayoshi Ichimiya, Masayoshi Ichimiya, Jun Miyake, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto JSAP-OSA Joint Symposia, JSAP 2014 2014年01月 [査読無し][通常論文]
  • Chikako Ninagawa, Hirohiko Niioka, Tsutomu Araki, Mamoru Hashimoto JSAP-OSA Joint Symposia, JSAP 2014 2014年01月 [査読無し][通常論文]
  • Doan T. Kim Dung, Shoichiro Fukushima, Hirohiko Niioka, Masayoshi Ichimiya, Masayoshi Ichimiya, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto, Jun Miyake JSAP-OSA Joint Symposia, JSAP 2014 2014年01月 [査読無し][通常論文]
  • Upconversion fluorescence and CL imaging for multiscale biological imaging
    H. Niioka, T. Furukawa, S. Fukushima, M. Ichimiya, J. Miyake, M. Ashida, T. Araki, M. Hashimoto Proceedings of 2013 Annual Meeting of the Spectroscopical Society of Japan 49 -49 2013年11月 [査読無し][通常論文]
  • Bimodal biological observation with luminescence emitted under electron beam and near-infrared light irradiation
    S. Fukushima, T. Furukawa, H. Niioka, M. Ichimiya, J. Miyake, M. Ashida, T. Araki, M. Hashimoto Proceedings of 2013 Annual Meeting of the Spectroscopical Society of Japan 90 -90 2013年11月 [査読無し][通常論文]
  • S. Fukushima, T. Furukawa, H. Niioka, M. Ichimiya, M. Ichimiya, T. Nagata, J. Miyake, M. Ashida, T. Araki, M. Hashimoto JSAP-OSA Joint Symposia 2013 2013年09月 [査読無し][通常論文]
  • Synthesis of Rare-earth Doped Nano Phosphors for Biological Cathodoluminescence Imaging
    T. Furukawa, H. Niioka, M. Ichimiya, S. Ichikawa, T. Nagata, J. Miyake, M. Ashida, T. Araki, M. Hashimoto Focus on Microscopy 2013 (Maastricht, The Netherlands) (Poster) 2013年03月 [査読無し][通常論文]
  • 福島 昌一郎, 古川 太一, 新岡 宏彦, 一宮 正義, 芦田 昌明, 三宅 淳, 荒木 勉, 橋本 守 バイオフロンティア講演会講演論文集 2013.24 (0) 181 -182 2013年 [査読無し][通常論文]
  • Rare-earth Doped Y2O3 Nanophosphors Synthesized for Bio-imaging with Using CL and Fluorescence Microscopy
    H. Niioka, T. Furukawa, M. Ichimiya, T. Nagata, M. Ashida, T. Araki, M. Hashimoto 8th Handai Nanoscience and Nanotechnology International Symposium 2012年12月 [査読無し][通常論文]
  • Multimodal Imaging via Light Microscopy and Cathodoluminescence Microscopy for Biological Specimens with Rare-earth Doped Nanophosphors
    T. Furukawa, H. Niioka, M. Ichimiya, T. Nagata, M. Ashida, T. Araki, M. Hashimoto JSAP-OSA joint Symposia 2012, the 73th Autumn Meeting 2012年09月 [査読無し][通常論文]
  • ナノ蛍光体粒子とカソードルミネッセンス顕微鏡を用いたマルチカラー生体イメージング
    新岡宏彦, 古川太一, 一宮正義, 芦田昌明, 荒木勉, 橋本守 日本顕微鏡学会 (つくば国際会議場, 2012/5/14-16, 招待講演) 2012年05月 [査読無し][招待有り]
  • T. Furukawa, H. Niioka, M. Ichimiya, T. Nagata, M. Ashida, T. Araki, M. Hashimoto Program and Abstract Book, Focus on Micrscopy 2012 (FOM2012) 2012年04月 [査読無し][通常論文]
  • 岩塚 純一, 南川 丈夫, 新岡 宏彦, 荒木 勉, 橋本 守 バイオエンジニアリング講演会講演論文集 2012 (0) _7D31 -1_-_7D31-2_ 2012年 [査読無し][通常論文]
  • 村上 貴親, 南川 丈夫, 荒木 勉, 橋本 守 バイオエンジニアリング講演会講演論文集 2012 (0) _7D46 -1_-_7D46-2_ 2012年 [査読無し][通常論文]
  • Super-resolution imaging of nano phosphors via cathodoluminescence microscopy for biological imaging
    H. Niioka, T. Furukawa, M. Ichimiya, M. Ashida, T. Araki, M. Hashimoto 2011年12月 [査読無し][通常論文]
  • 南川 丈夫, 荒木 勉, 橋本 守 オプトロニクス 28 (8) 103 -107 2009年08月 [査読無し][通常論文]
  • 南川 丈夫, 新岡 宏彦, 荒木 勉, 橋本 守 バイオフロンティア講演会講演論文集 2009 (0) 93 -94 2009年 [査読無し][通常論文]
  • 生体組織の無染色顕微鏡観察術
    橋本 守, 福島 修一郎, 安井 武史, 荒木 勉 光アライアンス Vol. 20, No. 3 24 -28 2009年 [査読無し][通常論文]
  • ビーム断面内偏光分布制御と顕微観測への応用
    橋本 守, 吉木 啓介, 荒木 勉 光アライアンス 20 (4) 21 -25 2009年 [査読無し][通常論文]
  • SHG イメージングとCARS イメージング
    荒木勉, 橋本守, 安井武史, 福島修一郎 ぶんせき No. 9 490 -495 2009年 [査読無し][通常論文]
  • 南川 丈夫, 橋本 守, 荒木 勉 関西支部講演会講演論文集 2008 (0) _6 -2_ 2008年 [査読無し][通常論文]
  • 橋本 守 光学 36 (3) 143 -148 2007年 [査読無し][通常論文]
  • 分子の3次元的な向きを観測する偏光分布制御顕微鏡
    橋本守, 吉木啓介, 荒木勉 レーザ加工学会誌 13 (2) 140 -143 2006年04月 [査読無し][通常論文]
  • 橋本 守, 浅田崇裕, 荒木 勉 分光研究 54 (1) 32 -34 2005年 [査読無し][通常論文]
  • 市村垂生, 早沢紀彦, 橋本守, 井上康志, 河田聡 日本分光学会講演要旨集 2004 56 2004年05月18日 [査読無し][通常論文]
  • 市村垂生, 早沢紀彦, 橋本守, 井上康志, 河田聡 応用物理学関係連合講演会講演予稿集 51st (3) 1229 2004年03月28日 [査読無し][通常論文]
  • 市村垂生, 早沢紀彦, 橋本守, 井上康志, 河田聡 応用物理学関係連合講演会講演予稿集 51st (3) 1142 2004年03月28日 [査読無し][通常論文]
  • 市村垂生, 早沢紀彦, 橋本守, 荒木勉, 井上康志, 河田聡 レーザ顕微鏡研究会講演会論文集 29th 6,7 2003年07月01日 [査読無し][通常論文]
  • 市村垂生, 早沢紀彦, 橋本守, 井上康志, 河田聡 応用物理学関係連合講演会講演予稿集 50th (3) 1107 2003年03月27日 [査読無し][通常論文]
  • 橋本 守 分光研究 49 (2) 51 -61 2000年 [査読有り][通常論文]
  • 同期式ナノ秒Xeアークランプ低ジッター化とその応用
    藤澤泰充, 橋本 守, 伊丹 伸, 荒木 勉 中国四国支部第35期総会・講演会講演論文集 975 (1) 325 -326 1997年 [査読無し][通常論文]
  • 橋本 守, 湯沢 哲朗, 浜口 宏夫 Jacso Report 37 (3) 27 -33 1995年06月 [査読有り][招待有り]

特許

  • 特許6817623:偏光制御装置および偏光制御方法    2021年01月20日
    吉木啓介, 橋本守  公立大学法人兵庫県立大学
  • 特許6425242:変調光検出のSN比を向上する方法    2018年11月21日
    橋本 守, 川人 祥二  国立大学法人静岡大学, 国立大学法人大阪大学
  • 特許6032574:スペクトル分解能とスペクトル確度を向上するフーリエ変換型分光法、分光装置および分光計測プログラム    2016年11月30日
    安井 武史, 橋本 守, 荒木 勉, 弥永 祐樹  国立大学法人大阪大学
  • 特許5984112:カソードルミネッセンス用標識試薬    2016年09月06日
    新岡 宏彦, 橋本 守, 古川 太一  国立大学法人大阪大学
  • 橋本 守, 丸山 真幸  国立大学法人大阪大学, 株式会社メガオプト
  • 特許5831901:誘導ラマン散乱顕微鏡    2015年12月09日
    橋本 守  国立大学法人大阪大学
  • 特許4862164:パルスレーザ光のタイミング調整装置,調整方法及び光学顕微鏡    2012年01月25日
    橋本 守, 南川 丈夫, 谷本 尚生, 小林 実, 藤田 克昌, 河田 聡, 荒木 勉  国立大学法人大阪大学  
    特願2006-135293

受賞

  • 2014年07月 国立大学法人 大阪大学 第3回 大阪大学総長顕彰 研究部門
     
    受賞者: 橋本守
  • 2010年02月 財団法人 中谷医工計測技術財団 中谷賞
     コヒーレントアンチスト―クスラマン散乱による生体分子の無染色な高解像・高速観測 
    受賞者: 橋本 守
  • 2009年08月 高速電子処理応用技術学会 優秀発表賞
     DSPを用いたピコ秒レーザーの高精度制御システムの開発 
    受賞者: 橋本 守
  • 2009年03月 社団法人 日本機械学会 2008年度日本機械学会船井賞
     テラヘルツ・カラースキャナーの開発 
    受賞者: 安井武史;橋本守;荒木勉
  • 2006年01月 財団法人 コニカミノルタ画像科学振興財団 コニカミノルタ画像科学奨励賞
     分子のベクトル場的分布画像を観測する光学顕微鏡システム 
    受賞者: 橋本 守
  • 2002年05月 社団法人 日本分光学会 日本分光学会賞論文賞
     コヒーレントアンチスト―クスラマン散乱を用いた顕微鏡 
    受賞者: 橋本 守

共同研究・競争的資金等の研究課題

  • 無侵襲血液成分モニタリングのための血中微粒子濃度の経皮光学計測技術の開発
    日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2020年04月 -2023年03月 
    代表者 : 加藤 祐次, 清水 孝一, 橋本 守, 松村 健太
  • 非線形ラマン散乱内視鏡実現のための4光波混合現象の削減法の開発
    公益財団法人 村田学術振興財団:研究助成
    研究期間 : 2020年06月 -2021年05月
  • ハイパースペクトル非線形ラマン散乱イメージングによる人工知能病理診断
    日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2017年04月 -2020年03月 
    代表者 : 橋本 守, 高松 哲郎, 加藤 祐次, 三宅 淳, 新岡 宏彦
     
    人工知能を用いた次々世代の病理診断およびスクリーニング技術を確立することを目的として,ハイパースペクトル非線形ラマン散乱イメージの人工知能を用いた解析により,病変部や正常部の分別,組織の分別する手法の確立を目指す. まずは,人工知能により観測データのノイズ低減による観測時間の短縮化について検討した.非線形ラマン散乱顕微鏡を用いて異なる露光時間によるCARS画像を観測し,長時間露光で観測した画像を教師データ,短時間露光で観測した画像を入力データとして深層学習を行った.また,様々な露光時間で観測した画像の画質を評価し,深層学習によって再構成した画像と比較することにより,深層学習によるノイズ低減効果を見積もった.実験結果から,深層学習を用いた再構成により,露光時間を10から15としたことに相当するイメージを取得できることがわかった.一般に広く用いられるガウシアンフィルターの場合には分解能の低下が見られるが,深層学習によれば分解能の低下を抑えながら,画質の向上をはかれることがわかった.これらの学習結果を非線形ラマン散乱硬性鏡イメージへと適用(転移学習)し,数秒の露光時間でも十分なSN比の観測が可能であることが示された. 蛍光画像により組織の分別の学習を行い,これを非線形ラマン散乱像へ転移学習することで分別精度が向上することも確かめた.非線形ラマン散乱像の取得は,蛍光観測に比べて観測時間を要するため大量のデータを取得することが難しいが,容易に大量な画像を取得することが容易な蛍光画像を用いて予備学習を行うことで,非線形ラマン散乱像の精度向上を目指すことができることがわかった.
  • 日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2015年04月 -2019年03月 
    代表者 : 福島 修一郎, 紀ノ岡 正博, 橋本 守
     
    細胞の培養環境を細胞スケールで制御することの重要性をマイクロ流体デバイスを用いて検証した.酸素濃度制御型デバイスにより,従来法では制御が困難な異なる酸素環境の細胞の共培養を実現することができた.また,細胞培養過程の非侵襲観測の方法として,非線形光学顕微鏡の有用性を示した.コラーゲンゲルを培養基質とする場合は,第2高調波発生光を用いた基質構造の可視化および変形解析が力学的環境要因の検討に有効である.
  • 日本学術振興会:科学研究費助成事業 挑戦的研究(萌芽)
    研究期間 : 2017年06月 -2019年03月 
    代表者 : 橋本 守
     
    光の回折限界を越える,非線形ラマン散乱と原子間力顕微鏡(AFM)とを組み合わせた超解像分子振動イメージング顕微鏡を開発した.非線形ラマン散乱過程を利用して特定の分子振動を励起し,この分子振動励起に伴う体積膨張をAFMで観測する.レーザー光をAFMのプローブ付近に集光するため,この影響を除去しながら高感度に体積膨張を検出する必要がある.そこで探針の試料接触の際のみに2台のパルス光が同時に試料に照射されるように,2台のレーザーのパルス発振の周波数差が,カンチレバーの共振と同期させるシステムの開発した.開発した装置を使って,非線形ラマン散乱によって誘起された体積膨張と思われる信号の検出に成功した.
  • 日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2016年04月 -2019年03月 
    代表者 : 三浦 治郎, 北條 裕信, 橋本 守, 久保 美寿穂, 荒木 勉
     
    長期的に糖尿病に罹患している人の歯髄内には顆粒状石灰化が認められ、病理組織学的には進行性病変の副産物として考えられている。しかし、周囲が硬組織かつ未脱灰での観察が必要であるため試料加工が難しく電子顕微鏡による超微形態観察の報告は少ない。本研究では、2型糖尿病モデルラットの臼歯の石灰化結石に対して、多面的な解析を行った。結果として、石灰化時期は糖尿病ラットにおいて10週齢前後=高値(1000mg/dl付近)あたりで歯髄内石灰化が観察され始めることが分かった。石灰化メカニズムを解明することで新規治療法の提案や全身疾患、加齢やう蝕に関連する歯の糖化現象についての詳細な研究を行った。
  • 日本学術振興会:科学研究費助成事業 挑戦的萌芽研究
    研究期間 : 2016年04月 -2018年03月 
    代表者 : 三浦 治郎, 橋本 守
     
    蛋白質による糖化反応は複雑で未解明の部分も多いが、最終的に非常に安定なAGE架橋がコラーゲン線維間に生成される. AGEs架橋が過度に形成されるとコラーゲン線維の色調や物性が変化する。同時にコラーゲンの自家蛍光の蛍光寿命がAGEsの産生に伴い短くなるという報告もある。AGEsの局在や組織への蓄積を光学的に分析することで組織の糖化レベルを評価ことができ、糖化現象が生体にあたえる影響を解明できそれらを応用することで生体老化の指標を策定できるのではないかと考えた。本研究で人及びラット組織のAGEs沈着状況と蛍光寿命の変化を詳細に測定し生体の糖化が蛍光寿命を基準として評価できる可能性が示唆された。
  • 日本学術振興会:科学研究費助成事業 基盤研究(A)
    研究期間 : 2014年04月 -2017年03月 
    代表者 : 安井 武史, 岩田 哲郎, 橋本 守, 木戸口 善行, 埴淵 昌毅
     
    本研究では、周期的な繰り返し現象を観測するフーリエ変換分光法において、観測時間窓と繰り返し周期とを厳密に一致させて時間波形を取得し、その時間波形をフーリエ変換することにより、無限小スペクトル分解の離散フーリエ変換スペクトルが取得可能であることを確認した。さらに、繰り返し周期の走査で離散スペクトルを高密度化することにより、フーリエ変換分光法の分光性能を大幅に向上出来ることを、テラヘルツ領域及び近赤外領域の分光計測で実証した。
  • 高速誘導ラマン散乱スペクトルイメージングシステムの開発
    科学技術振興機構:産学共創基礎基盤研究プログラム「ヒト生体イメージングを目指した革新的バイオフォ トニクス技術の構築」
    研究期間 : 2010年12月 -2017年03月 
    代表者 : 橋本 守
  • 日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2013年04月 -2016年03月 
    代表者 : 橋本 守, 福島 修一郎, 新岡 宏彦, 新岡 宏彦
     
    4次ラマン散乱(FOCRS)分光顕微鏡を結晶化モニタリングに用いることを提案した.結晶化は飽和溶液中でのプロセスであり,結晶と飽和溶液からの信号を分別する必要がある.偶数次の非線形光学現象は,中心対称性を持つ物質からは禁制となるため,4次の非線形光学効果であるFOCRSを用いることで,非中心対称性の結晶の分子振動情報の,溶液の影響ない観測が期待できる. FOCRS分光顕微鏡を開発し,対称心を持つ水和DAST結晶と対称心を持たない無水DAST結晶のFOCRSとCARSを観測した.CARSでは両者でラマンバンドが得られるが,FOCRSでは無水結晶のみラマンバンドを示し,FORCSの選択性を示した.
  • 日本学術振興会:科学研究費助成事業 基盤研究(A)
    研究期間 : 2011年04月 -2014年03月 
    代表者 : 荒木 勉, 橋本 守, 福島 修一郎, 新岡 宏彦
     
    複雑系の中から所定の生体分子の形態・高次構造および変化をイメージングする手法の開発と応用を目指す。ここでは第二高調波発生光顕微鏡(SHG顕微鏡)とカソードルミネッセンス(CL)顕微鏡について研究した。SHG顕微鏡では人の皮膚をin vivo測定し、光老化におけるSHG光強度の変化を追跡した。また本方法を培養細胞のコロニー状況観測に利用し、さらにコラーゲンゲルと細胞との機械的な相互干渉を可視化した。 CL顕微鏡では10 nmレベルの空間分解で観察できうるナノ蛍光体作製に成功した。また、ナノ蛍光体を細胞内に取り込ませ、細胞内においてもCLイメージングに成功した。
  • カソードルミネッセンス顕微鏡による細胞中の高空間分解能蛋白質イメージング
    立石科学技術振興財団:
    研究期間 : 2011年04月 -2013年03月 
    代表者 : 橋本 守
  • 日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2010年 -2012年 
    代表者 : 橋本 守, 福島 修一郎, 新岡 宏彦
     
    多焦点コヒーレントアンチストークスラマン散乱顕微鏡とレーザーアブレーション法を組み合わせ,レーザー光による遠隔的な刺激に対する試料の応答をリアルタイム・無染色に観測可能なシステムを構築した.アブレーション用レーザー光は,観測システムとは独立にその照射位置をコンピュータ制御可能である.開発した装置を,生細胞の細胞膜破壊に対する応答観測,ならびにレーザー放射圧による結晶化プロセスの観測へ応用した
  • 日本学術振興会:科学研究費助成事業 基盤研究(A)
    研究期間 : 2008年 -2010年 
    代表者 : 荒木 勉, 橋本 守, 福島 修一郎, 紀ノ岡 正博, 岩田 哲郎, 安井 武史
     
    極短パルスレーザーによって誘起される非線形光学現象を利用して、無染色で生体分子を可視化することができる生体分子イメージング顕微鏡を開発し、これらによって実際の細胞や生体組織を観察し生理活性に関する情報の取得することを目的としている。具体的にはSHG顕微鏡とCARS顕微鏡を対象とし、初年度は装置基盤技術の確立を進め、次年度以降はその改良を通じて実用機器の開発を目指すとともに、ヒト真皮コラーゲンのin situ測定や生細胞のリアルタイム計測ならびに応用や目指した。
  • 日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2007年 -2009年 
    代表者 : 橋本 守, 福島 修一郎, 藤田 克昌, 吉木 啓介
     
    ラマンイメージングの実時間観測を目指して,多焦点非線形ラマン散乱顕微鏡を開発した.2台のpsレーザーを高精度に同期させ(ジッター30fs),かつ波長を300msで走査可能なレーザーシステムを開発し光源に用いた.生細胞の実時間観測(100ms/image),レーザーアブレーションによる細胞膜の破壊とその修復過程の観測等が可能であることを示した.また,膜たんぱく質の選択的な観測を目指して,集光スポットでの電磁場の向きを自在に制御するシステムを構築し,液晶分子の配向観測によってその可能性を示した.
  • サブミクロンの分解能で液晶分子の 3 次元配向を観測する顕微鏡
    新エネルギー・産業技術総合開発機構:平成 17 年度 第 2 回産業技術研究助成事業
    研究期間 : 2006年01月 -2008年12月 
    代表者 : 橋本 守
  • 日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2007年 -2008年 
    代表者 : 井上 康志, VERMA Prabhat, 橋本 守, 藤田 克昌, 橋本 守, 藤田 克昌
     
    金属ナノ構造に誘起される局在プラズモンは、光(フォトン)と共鳴的に結合することで、光をナノ領域に閉じ込め、さらに光電場を増強する。本研究では、この近接場ナノプラズモニクス技術を駆使することで、距離に強く依存した分子-金属原子間の相互作用を計測するナノラマン分光装置を開発し、さらにこれを用いて、距離に応じて分子とナノ探針先端の金属原子の間に電磁気学的、化学的、さらに力学的な相互作用が各々働くことを明らかにした。
  • リアルタイム CARS 顕微鏡
    稲盛財団:平成 18 年度稲盛財団研究助成金
    研究期間 : 2006年04月 -2007年03月 
    代表者 : 橋本 守
  • テラヘルツ光照射に対する細胞応答のマルチフォトニクス・モニタリング
    日本学術振興会:科学研究費助成事業 萌芽研究
    研究期間 : 2006年 -2007年 
    代表者 : 安井 武史, 福島 修一郎, 橋本 守, 荒木 勉, 東野 義之
     
    過渡的テラヘルツ光照射に対する細胞応答を観察する手段として利用する非同期光サンプリング式テラヘルツ・ポンプ・プローブ法において、テラヘルツ・スペクトルの広帯域化を実現するため、2台の極超短パルスレーザー(パルス幅10fs)を用いた非同期光サンプリング光源を開発した。また、2台のレーザーのモード同期周波数及び両レーザーのモード同期周波数差の高安定化に取り込み、従来光源と比較して10倍以上の高安定化を達成した。そして、本光源を用いた非同期光サンプリング式テラヘルツ時間領域分光装置で低圧水蒸気ガスのスペクトル計測を行い、本装置のスペクトル分解能が7GHzであることを確認した。テラヘルツ・ポンプ・プローブ法で細胞応答を観察するためにはスペクトル分解能がまだ不足しており、レーザー制御手法を再検討し、さらなる高分解能化をはかる必要がある。 定常的テラヘルツ照射に対する細胞内応答の観察のために、アクティブ周波数逓倍器と周波数シンセサイザーを用いた連続発振(CW)テラヘルツ光源を開発し、パワー10mW、スペクトル線幅1Hz以下、周波数安定度10-11、チューニング範囲75GHz〜110GHzの基本特性を得た。一方、生体SHG(第2高調波発生光)イメージングは細胞内コラーゲン産生に関する情報を抽出するために利用する。今年度は、深部プローブ型SHGイメージング装置(レーザー波長1250nm)及び高分解SHGイメージング装置(レーザー波長800nm)を様々な生体サンプルに適用し、その有用性を評価した。その結果、細胞培養コラーゲンゲル、光老化真皮、心臓弁膜症によるコラーゲン変性、軟骨表層部、などの観察に有用であることが分かった。
  • 擬似ランダム符号を用いたパルスシェーピング多焦点CARS顕微鏡
    日本学術振興会:科学研究費助成事業 萌芽研究
    研究期間 : 2006年 -2007年 
    代表者 : 橋本 守
     
    生きたままの生体組織・細胞を構成する分子ならびにその機能を観測する"分子イメージング"技術は,ポストゲノム研究において必要不可欠な研究ツールとなる.CARS(コヒーレントアンチストークスラマン散乱)顕微鏡は,全ての分子が持つ分子振動を観測することにより,無染色に分子種ならび分子構造情報を得ることができ,また非線形光学効果による回折限界を超えた空間分解能と誘導放出過程による高い感度を備えた観測手段である.しかしながら,これまでのCARS顕微鏡では2色のレーザーを必要としていたために,装置が複雑なものとなっていた。本研究では,一台の超短パルスレーザーを使用し,パルスシェーピングすることによって選択的に分子振動を励起する手法の開発を行った. 昨年度,作製したパルスシェーピング装置を改造し,2ビームを同時にパルスシェーピング可能なえるものとした.これにより,相互相関を容易に観測できるようになった.パルスシェーピング時に与える変調方法の比較を行った.正弦波状の位相変調を与えるものと,擬似ランダム符号を与えた時との比較を行ったところ,擬似ランダム符号を与えた方が,相互相関信号のパルス列の数が多くなる(正弦波では3-5本,擬似ランダム符号では9-13本)ことが確認できた.また,四塩化炭素を試料にCARS信号を観測した結果,分子振動が選択的に励起(周波数分解能が約3倍に向上)されていることが確認できた.
  • 無染色・生体分子イメージング顕微鏡の開発と培養細胞の品質制御への応用
    日本学術振興会:科学研究費助成事業 基盤研究(A)
    研究期間 : 2005年 -2007年 
    代表者 : 荒木 勉, 橋本 守, 紀ノ岡 正博, 安井 武史, 岩田 哲郎, 東野 義之
     
    無染色・生体分子イメージング顕微鏡の開発と培養細胞の品質制御への応用を目的としている。初年度は要素技術・基盤技術の確立を進め、次年度はその改良を通じて実用機器の開発を目指した。さらに培養細胞の品質制御への応用を目指した。最終年度には、開発機器の評価と応用を進め、その研究成果を論文にまとめる作業を行った。具体的な成果は以下の通りである。 (1)真皮コラーゲンの高速経皮測定を目的とし、高機能SHG顕微鏡を開発した。その結果、表皮から深さ300ミクロンまでのコラーゲンの断層像を得ることた成功した。また、心臓弁など各種組織のコラーゲン配向像の取得に成功した。断層画像1枚あたりの取得時間は従来は1分程度であったが1秒にまで短縮した。 (2)SHG発生分子の3次元方向ベクトルを同定できるラジアル偏光を用い、生体分子の3次元配向イメージングに成功した (3)細胞培養の培地としてのコラーゲンに着目し、細胞の増殖によってコラーゲンゲルの分子配向が変化していく様子をSHG顕微鏡によってはじめて追跡するなど、品質管理に関する研究を行った。 (4)生体分子を無染色で同定するためのCARS顕微鏡について、レーザー間のタイミングを高精度で制御し、かつマルチビーム測定を行うことでビデオレートのリアルタイム測定を試みた。本顕微鏡によってリポゾームの形態変化の様子を捉えることが出来た。 (5)原子発光分析装置を駆使して加齢と生体元素間の相関を調査した。また、力学的所見から局所のカルシウム沈着についての解析を行った。 (6)時間分解蛍光測光に関して、カルシウムイオン定量への応用を行い、また歯の加齢との相関を求めた。さらに歪んだ波形から真の蛍光寿命を求めるための新しい手法を開発した。
  • 分子のベクトル分布観測顕微鏡の開発
    伊藤科学振興会:平成 17 年度研究助成
    研究期間 : 2005年09月 -2006年08月 
    代表者 : 橋本 守
  • 2 光子検出器を用いた二台のピコ秒モードロックレーザーの高精度同期
    村田学術振興財団:第 21 回 (平成 17 年度) 研究助成
    研究期間 : 2005年06月 -2006年08月 
    代表者 : 橋本 守
  • ピコ秒レーザーの自動安定発振・パルス長最適化機構の開発
    科学技術振興機構:シーズ育成試験
    研究期間 : 2006年01月 -2006年03月 
    代表者 : 橋本 守
  • 非線形光学効果による赤外発生とプローブレス近接場赤外顕微鏡への展開”
    日本学術振興会:若手研究 A
    研究期間 : 2003年04月 -2005年03月 
    代表者 : 橋本 守
  • ファン・デル・ワールス力制御ラマン散乱ナノ分光法
    日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2004年 -2005年 
    代表者 : 井上 康志, 橋本 守, 藤田 克昌
     
    ナノ材料の代表格であるカーボンナノチューブ(CNT)にファン・デル・ワールス力を印加することで特定のラマンバンドだけがシフトする現象を見出し、解析した。具体的には、単一のCNTバンドルに、原子間力顕微鏡用のカンチレバーに銀をコーティングした金属ナノ探針で応力を印加しながらチップ増強近接場ラマンスペクトルを測定し、CNTのラマンバンドの変化をin situで観察した。応力を0nNから2.4nNまで段階的に印加していくと、CNTの電気的性質を反映するG-bandと呼ばれるラマンバンドに対応する1600cm^<-1>付近に観測される2つのピークのうち、低波数側のピークは18cm^<-1>だけ低波数側にシフトした。一方、高波数側のピークは変化しなかった。この現象は、2つのピークが振動方向の異なる振動モードに対応し、それぞれの振動方向によってせん断応力が異なることに起因すると考えている。さらに、200cm^<-1>〜300cm^<-1>付近に観測されるCNTの直径に依存した振動モードであるRadial Breathing Modeのピークは最大で5cm^<-1>だけ低波数側にシフトすることを観察した。この低波数シフトは、探針によって一軸性の応力がカーボンナノチューブに印加されたことにより、カーボンナノチューブの断面が楕円形に歪んだことに起因すると考えている。また、応力を印加するとすべてのラマンバンドのラマン散乱光強度が増幅することも観察した。これはCNTが直径方向に歪むことで、バンドギャップエネルギーが共鳴エネルギー(2.33eV)に近づき、共鳴ラマン効果の寄与が大きくなったことを示唆している。 さらに、複数の金属原子により形成されるクラスターと分子とによる錯体の振動計算を密度汎関数法により行った。その結果、銀原子数が4つ以上あれば、錯体形成によるバンドシフトが実験値にほぼ一致することを確かめた。
  • 3次元偏光制御顕微鏡による3次元分子配向の3次元観測
    日本学術振興会:科学研究費助成事業 萌芽研究
    研究期間 : 2004年 -2005年 
    代表者 : 橋本 守, 安井 武史, 福島 修一郎
     
    従来の光学顕微鏡技術では,高い空間分解能を持ちながら3次元的な分子の配向を観測することは不可能であった.これは,通常の直線偏光ビームを集光しても焦点での電場の向きは光軸に対して直行しているために,光軸方向に向いた分子を観測することはできないためである.一方,ラジアル偏光と呼ばれる放射状の偏光を高い開口数を持つ対物レンズで集光すると焦点で光軸方向の電場が形成されるために光軸方向に向いた分子も観測することができることが知られている.したがって,レーザー走査顕微鏡においてレーザービーム断面内の偏光分布を自由に制御し,直線偏光やラジアル偏光を高速に変化させることができれば焦点の電場の方向を3次元的に変化させることができる.また,この偏光分布制御と非線形光学効果を組み合わせることで,6次元観測(3次元空間+3次元配向)することが可能となる. フェムト秒超短パルスレーザー光を,平行配向した液晶からなる液晶空間位相変調素子(SLのM)を2回通すことにより,任意の空間的偏光分布を形成し,この光を励起光に用いた第二高調波発生顕微鏡を構築した.SLMに印加するパターンを変え,直交した一様な偏光直線偏光ビームと,ラジアル偏光を試料に入射し,その第二高調波発生を観測することで,試料の分子配向を観測することが可能かどうか検証した.試料には,コラーゲンを主成分とし,コラーゲン線維が一方向に並んだヒトアキレス腱を用いた.コラーゲンは線維方向と入射電場の方向が一致した場合に強い第二高調波を発生することが知られているが,光軸方向にコラーゲン線維が向いた試料を開発した装置で観測したところ,ラジアル偏光励起で強い信号が得られ,提案手法の原理確認を行うことができた.
  • 極短パルスレーザーによる生体分子認識イメージング法の開発と組織診断への応用
    日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2003年 -2004年 
    代表者 : 荒木 勉, 橋本 守, 安井 武史, 東野 義之, 岩田 哲郎
     
    複雑系の中から所定の生体分子を非侵襲・非染色、生きたまま、かつリアルタイムで認識し、その形態・高次構造および変化をイメージングする技術の開発を目的としている。そこで我々は、高速レーザー誘起で現れる生体組織のフォトニクス現象を総合し、ミクロレベルでの細胞・組織の診断に利用することを試みた。具体的にはナノ秒蛍光と第二高調波光(SHG光)および非線形ラマン分光法に着目した。この中でフェムト秒レーザー光によってSHG光がコラーゲンから特異的に発生することに注目し、in situでコラーゲン分子の配向分布画像を観測できる顕微鏡システムを開発した。この成果の概要を記すと次のようになる。(1)フェムト秒レーザー光(波長810nm)をコラーゲン組織に照射し、波長405nmのSHG光を発生させた。(2)偏光レーザービームで誘起されたSHG光を検光子を通じて検出することで、線維配向の方位分布を知ることができた。(3)奥行き方向20μmの分解を持つ共焦点光学系顕微鏡システムを構築し、コラーゲン配向の3次元断層イメージング取得に成功した。(4)この計測システムを用いて、ヒト海綿骨および経皮測定による真皮のコラーゲン配向計測を試み、各組織のコラーゲン分子の配向方位マップ取得に成功した。次にナノ秒蛍光寿命分布画像取得システムを試作した。ここで得られた結果から、SHGと蛍光の情報を合わせて生体診断に寄与できるとの確証を得た。さらに非線形ラマン分光画像においては赤血球の周辺に存在するヒアルロン酸の分布像取得に成功した。また、フェムト秒レーザーを血糖値計測へ応用する試みに着手し、基本手法を論文として報告した。
  • 非線形光学効果による赤外発生とプローブレス近接場赤外顕微鏡への展開
    日本学術振興会:科学研究費助成事業 若手研究(A)
    研究期間 : 2003年 -2004年 
    代表者 : 橋本 守
     
    近年のナノテクノロジーの発達に伴い,高い空間分解能を持ちながら微小物体の組成や分子構造を観測する手法の開発が望まれている.すべての分子は分子振動を有し,その周波数は分子構造に非常に敏感であるため,分子振動を観測する振動分光による高空間分解能観測に関する研究を行った. 赤外光をそのまま集光しても回折限界により波長程度までしか集光できないために,赤外顕微鏡の空間分解能は10ミクロン程度に限定される.しかしながら,可視あるいは近赤外光を用いて赤外光を発生させることができたなら,可視光あるいは近赤外光の波長程度の空間分解能を持った顕微鏡を構成することができる.大気中でレーザー光を集光して誘電破壊(レーザーブレイクダウン)により発生するマイクロプラズマを光源とした近接場顕微鏡の開発を目指して,マイクロプラズマの発生,観測に関する研究を行った.高強度なフェムト秒レーザーをNA=0.6の対物レンズで強く集光することにより,直径2ミクロン程度のマイクロプラズマを生成することに成功した.また,このプラズマから1000cm^<-1>から4000cm^<-1>に渡る赤外光が放射されていることを実験的に確認した.しかし,本手法を近接場へと展開した場合,プラズマにより試料が破壊されてしまった.そこで,金属薄膜での非線形光学効果を利用した波長変換技術を用いることを検討し,金属薄膜での差周波発生の確認を行った. また,コヒーレントアンチストークスラマン散乱(CARS)と呼ばれる非線形光学現象を,近接場顕微鏡へと適用した.これは,金属プローブ先端付近での電場増強効果と非線形光学効果による相乗効果により従来にない高空間分解能化した振動分光を行うことであり,波長約800nmの光を用いて15nmの空間分解能と,回折限界の数十分の1まで向上させることに成功した.光を用いた振動分光において従来にない空間分解能を達成することができた.
  • 紫外励起 CARS 顕微鏡のシステムの開発と生物組織観察への応用
    日本学術振興会:基盤 B 展開
    研究期間 : 2001年04月 -2003年03月 
    代表者 : 橋本 守
  • 多焦点 CARS 顕微鏡による 3 次元分子分布イメージング
    日本学術振興会:奨励研究 A
    研究期間 : 2001年04月 -2003年03月 
    代表者 : 橋本 守
  • コヒーレントアンチストークスラマン散乱による生体組織の3次元局所空間分子分光分析
    仲谷電子計測技術財団:第16回研究助成
    研究期間 : 2000年04月 -2001年03月 
    代表者 : 橋本 守
  • コヒーレントアンチストークスラマン散乱顕微鏡の開発と生体計測への応用
    日本学術振興会:奨励研究 A
    研究期間 : 1999年04月 -2001年03月 
    代表者 : 橋本 守
  • コヒーレントアンチストークスラマン散乱顕微鏡による生体試料の3次元イメージング
    住友財団基礎科学研究助成:
    研究期間 : 1999年11月 -2000年10月 
    代表者 : 橋本 守

教育活動情報

主要な担当授業

  • システム工学概論
    開講年度 : 2019年
    課程区分 : 学士課程
    開講学部 : 工学部
  • 医用システム工学特論
    開講年度 : 2019年
    課程区分 : 修士課程
    開講学部 : 情報科学研究科
    キーワード : 医用システム,医用計測,医用画像,診断・治療機器,医用情報,医用安全,生体医工学.
  • 医用システム工学特論
    開講年度 : 2019年
    課程区分 : 修士課程
    開講学部 : 情報科学院
    キーワード : 医用システム,医用計測,医用画像,診断・治療機器,医用情報,医用安全,生体医工学.
  • バイオエンジニアリング特論
    開講年度 : 2019年
    課程区分 : 修士課程
    開講学部 : 情報科学研究科
    キーワード : 遺伝情報, genetic information, バイオインフォマティクス, bioinformatics, イメージング, imaging, 生体医工学, biomedical engineering, 細胞力学, cell mechanics
  • 医用システム工学特論
    開講年度 : 2019年
    課程区分 : 博士後期課程
    開講学部 : 情報科学研究科
    キーワード : 医用システム,医用計測,医用画像,診断・治療機器,医用情報,医用安全,生体医工学.
  • 医用システム工学特論
    開講年度 : 2019年
    課程区分 : 博士後期課程
    開講学部 : 情報科学院
    キーワード : 医用システム,医用計測,医用画像,診断・治療機器,医用情報,医用安全,生体医工学.
  • バイオエンジニアリング特論
    開講年度 : 2019年
    課程区分 : 博士後期課程
    開講学部 : 情報科学研究科
    キーワード : 遺伝情報, genetic information, バイオインフォマティクス, bioinformatics, イメージング, imaging, 生体医工学, biomedical engineering, 細胞力学, cell mechanics
  • 応用光学
    開講年度 : 2019年
    課程区分 : 学士課程
    開講学部 : 工学部
    キーワード : 光、幾何光学、レンズ、顕微鏡、望遠鏡、干渉計
  • 応用光学Ⅰ
    開講年度 : 2019年
    課程区分 : 学士課程
    開講学部 : 工学部
    キーワード : 光、幾何光学、レンズ、顕微鏡、望遠鏡、干渉計
  • 応用数学Ⅱ
    開講年度 : 2019年
    課程区分 : 学士課程
    開講学部 : 工学部
    キーワード : 常微分方程式、偏微分方程式、ラプラス変換、フーリエ級数

大学運営

委員歴

  • 2020年01月 - 現在   レーザ顕微鏡研究会 監査
  • 2019年04月 - 現在   日本分光学会 北海道支部   支部長
  • 2019年01月 - 現在   日本光学会北海道支部   幹事
  • 1998年 - 現在   レーザ顕微鏡研究会   幹事   レーザ顕微鏡研究会
  • 2005年 - 2016年09月   日本分光学会 関西支部   幹事   日本分光学会 関西支部
  • 2007年01月 - 2008年12月   日本光学会   光科学及び光技術調査委員会 委員長   日本光学会


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