Researcher Profile and Settings

Affiliation

• Faculty of Information Science and Technology　Bioengineering and Bioinformatics　Bioengineering

Job Title

• Professor

Degree

• Ph. D（1997/06 Univ. of Tokyo）

URL

https://www.ist.hokudai.ac.jp/labo/bmsys/index.html

J-Global ID

• 200901045896159714

Research Interests

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

Research Areas

• Nanotechnology/Materials / Optical engineering and photonics
• Nanotechnology/Materials / Basic physical chemistry
• Life sciences / Biomaterials
• Life sciences / Biomedical engineering

Educational Organization

•  Bachelor's degree program　School of Engineering
•  Master's degree program　Graduate School of Information Science and Technology
•  Doctoral (PhD) degree program　Graduate School of Information Science and Technology

Academic & Professional Experience

• 2019/04 - Today Hokkaido University Division of Bioengineering and Bioinformatics, Faculty of Information Science and Engineering professor
• 2016/10 - 2019/03 Hokkaido University Graduate School of Information Science and Technology Division of Bioengineering and Bioinformatics Professor
• 2007/04 - 2016/09 Osaka University Graduate School of Engineering Science Department of Mechanical Science and Bioengineering Division of Bioengineering Assoc. Professor
• 2003/04 - 2007/03 Osaka University Graduate School of Engineering Science Department of Mechanical Science and Bioengineering Division of Bioengineering
• 2002/11 - 2003/03 Osaka University Graduate School of Engineering Science Assoc. Professor
• 1999/04 - 2002/10 Osaka University Graduate School of Engineering Science Department of Systems Innovation Division of Mathematical Science
• 1997/08 - 1999/03 Osaka University Graduate School of Engineering Science
• 1996/04 - 1997/07 The University of Tokushima Faculty of Engineering
• 1991/04 - 1996/03 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

Education

• 1997/06 -   The University of Tokyo
• 1989/04 - 1991/03  Osaka University  Graduate School of Engineering  Division of Applied Physics
• 1985/04 - 1989/03  Osaka University  School of Engineering

Association Memberships

• 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   

Research Activities

Published Papers

• A CMOS Double-Demodulation Lock-In Amplifier for Stimulated Raman Scattering Signal Detection

  Shukri Bin Korakkottil Kunhi Mohd, De Xing Lioe, Keita Yasutomi, Keiichiro Kagawa, Mamoru Hashimoto, Shoji Kawahito

  Electronics 12 (1) 4 - 4 2022/12/20 [Refereed]
   

  In typical stimulated Raman scattering (SRS) signal extraction, the photodetector and lock-in amplifier are often based on separate platforms, rendering the system cumbersome and non-scalable. This paper proposes an SRS double-demodulation lock-in amplifier implemented with a complementary metal-oxide semiconductor (CMOS) image sensor technology that integrates two-stage 1/f noise and offset reduction circuits with a high-speed lateral electric field modulation (LEFM) photo-demodulator. A weak SRS signal is buried in a large offset with a ratio of 10−4 to 10−6; boosting such signals in a CMOS device requires an extremely high offset and noise reduction capability. The double-modulation two-stage lock-in amplifier demodulates at 40 MHz with a sampling frequency of 20 MHz, can suppress the laser and circuit’s 1/f noise to achieve higher detection sensitivity. A prototype chip fabricated using 0.11 μm CMOS image sensor technology is evaluated. Both simulation and measurement results are presented to verify the functionality and show that the differential readout structure can successfully reject laser common mode components while emphasizing its differences. The measurement results show that the double-modulation lock-in amplifier effectively suppresses the circuit’s 1/f noise by a factor of nearly two decades.
• Near real-time nerve visualization using coherent Raman scattering rigid endoscope and deep learning-based image processing for nerve-sparing surgery

  Naoki Yamato, Hirohiko Niioka, Jun Miyake, Mamoru Hashimoto

  Biomedical Vibrational Spectroscopy 2022: Advances in Research and Industry 2022/03/02 [Refereed]
  
• Ultrahigh-speed multiplex coherent anti-Stokes Raman scattering microspectroscopy using scanning elliptical focal spot

  Shun Kizawa, Mamoru Hashimoto

  The Journal of Chemical Physics 155 (14) 144201 - 144201 0021-9606 2021/10/14 [Refereed][Not invited]
  
• Avoidance of four-wave mixing in optical fiber bundle for coherent anti-Stokes Raman scattering endomicroscopy

  Hiroki Ogawa, Mamoru Hashimoto

  Optics Letters 46 (14) 3356 - 3356 0146-9592 2021/07/15 [Refereed]
  
• Improvement of nerve imaging speed with coherent anti-Stokes Raman scattering rigid endoscope using deep-learning noise reduction

  Naoki Yamato, Hirohiko Niioka, Jun Miyake, Mamoru Hashimoto

  Scientific Reports 10 (1) 15212  2020/09 [Refereed][Not invited]
   

  A 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.
• Nerve Segmentation with Deep Learning from Label-Free Endoscopic Images Obtained Using Coherent Anti-Stokes Raman Scattering

  Naoki Yamato, Mana Matsuya, Hirohiko Niioka, Jun Miyake, Mamoru Hashimoto

  Biomolecules 10 (7) 1012 - 1012 2020/07/08 [Refereed][Not invited]
   

  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.
• Label-free nerve imaging with a coherent anti-Stokes Raman scattering rigid endoscope using two optical fibers for laser delivery

  Keigo Hirose, Shuichiro Fukushima, Taichi Furukawa, Hirohiko Niioka, Mamoru Hashimoto

  APL Photonics 3 (9) 092407  2018/07 [Refereed][Invited]
  
• Coherent anti-stokes Raman scattering rigid endoscope toward robot-assisted surgery

  K. Hirose, T. Aoki, T. Furukawa, S. Fukushima, H. Niioka, S. Deguchi, M. Hashimoto

  Biomedical Optics Express 9 (2) 387 - 396 2156-7085 2018/02/01 [Refereed][Not invited]
   

  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.
• Label-Free Biomedical Imaging Using High-Speed Lock-In Pixel Sensor for Stimulated Raman Scattering

  Kamel Mars, De Xing Lioe, Shoji Kawahito, Keita Yasutomi, Keiichiro Kagawa, Takahiro Yamada, Mamoru Hashimoto

  SENSORS 17 (11) 2581  1424-8220 2017/11 [Refereed][Not invited]
   

  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.
• 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 日本工業出版 28 (1) 24 - 28 0917-026X 2017/01 [Not refereed][Invited]
  
• Photo-Induced Cell Damage Analysis for Single- and Multifocus Coherent Anti-Stokes Raman Scattering Microscopy

  Takeo Minamikawa, Yoshinori Murakami, Naokazu Matsumura, Hirohiko Niioka, Shuichiro Fukushima, Tsutomu Araki, Mamoru Hashimoto

  JOURNAL OF SPECTROSCOPY 2017 5725340-1 - 5725340-8 2314-4920 2017 [Refereed][Not invited]
   

  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.
• Influence of Nonenzymatic Glycation in Dentinal Collagen on Dental Caries

  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 0022-0345 2016/12 [Refereed][Not invited]
   

  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.
• Multispectral Emissions of Lanthanide-Doped Gadolinium Oxide Nanophosphors for Cathodoluminescence and Near-Infrared Upconversion/Downconversion Imaging

  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 2079-4991 2016/09 [Refereed][Not invited]
   

  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).
• Correlative near-infrared light and cathodoluminescence microscopy using Y2O3:Ln, Yb (Ln = Tm, Er) nanophosphors for multiscale, multicolour bioimaging

  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  2045-2322 2016/05 [Refereed][Not invited]
   

  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.
• A Stimulated Raman Scattering CMOS Pixel Using a High-Speed Charge Modulator and Lock-in Amplifier

  De Xing Lioe, Kamel Mars, Shoji Kawahito, Keita Yasutomi, Keiichiro Kagawa, Takahiro Yamada, Mamoru Hashimoto

  SENSORS 16 (4) 532-1 - 532-16 1424-8220 2016/04 [Refereed][Not invited]
   

  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.
• Synthesis of Y2O3 nanophosphors by homogeneous precipitation method using excessive urea for cathodoluminescence and upconversion luminescence bioimaging

  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 2159-3930 2016/03 [Refereed][Not invited]
   

  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
• A CMOS Image Sensor Using High Speed Lock-In Pixels for Stimulated Raman Scattering

  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  0277-786X 2016 [Refereed][Not invited]
   

  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.
• Near-infrared broadband dual-frequency-comb spectroscopy with a resolution beyond the Fourier limit determined by the observation time window

  Sho Okubo, Yi-Da Hsieh, Hajime Inaba, Atsushi Onae, Mamoru Hashimoto, Takeshi Yasui

  OPTICS EXPRESS 23 (26) 33184 - 33193 1094-4087 2015/12 [Refereed][Not invited]
   

  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
• Orientation detection of a single molecule using pupil filter with electrically controllable polarization pattern

  Mamoru Hashimoto, Keisuke Yoshiki, Makoto Kurihara, Nobuyuki Hashimoto, Tsutomu Araki

  OPTICAL REVIEW 22 (6) 875 - 881 1340-6000 2015/12 [Refereed][Not invited]
   

  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.
• High-sensitivity and high-spatial-resolution imaging of self-assembled monolayer on platinum using radially polarized beam excited second-harmonic-generation microscopy

  Mamoru Hashimoto, Hirohiko Niioka, Koichiro Ashida, Keisuke Yoshiki, Tsutomu Araki

  APPLIED PHYSICS EXPRESS 8 (11) 112401  1882-0778 2015/11 [Refereed][Not invited]
   

  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
• Super-resolution discrete Fourier transform spectroscopy beyond time-window size limitation using precisely periodic pulsed radiation

  Takeshi Yasui, Yuki Iyonaga, Yi-Da Hsieh, Yoshiyuki Sakaguchi, Francis Hindle, Shuko Yokoyama, Tsutomu Araki, Mamoru Hashimoto

  Optica 2 (5) 460 - 460 2334-2536 2015/05/20 [Refereed][Not invited]
   

  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.
• Rare-earth-doped nanophosphors for multicolor cathodoluminescence nanobioimaging using scanning transmission electron microscopy

  Taichi Furukawa, Shoichiro Fukushima, Hirohiko Niioka, Naoki Yamamoto, Jun Miyake, Tsutomu Araki, Mamoru Hashimoto

  JOURNAL OF BIOMEDICAL OPTICS 20 (5) 056007-1 - 056007-6 1083-3668 2015/05 [Refereed][Not invited]
   

  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)
• Decrease in fluorescence lifetime by glycation of collagen and its application in determining advanced glycation end-products in human dentin

  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 2156-7085 2015/05 [Refereed][Not invited]
   

  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
• Multimodal Imaging Probing Platform Based on Upconverting Rare-Earth Doped Gd2O3 Nanocrystals

  Kim Dung T. Doan, Shoichiro Fukushima, Hirohiko Niioka, Masayoshi Ichimiya, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto, Jun Miyake

  BIOPHYSICAL JOURNAL 108 (2) 171A - 172A 0006-3495 2015/01 [Refereed][Not invited]
  
• Y2O3:Tm,Yb nanophosphors for correlative upconversion luminescence and cathodoluminescence imaging

  Shoichiro Fukushima, Taichi Furukawa, Hirohiko Niioka, Masayoshi Ichimiya, Jun Miyake, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto

  MICRON 67 90 - 95 0968-4328 2014/12 [Refereed][Not invited]
   

  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.
• Realtime cellar response observation by multi-focus CARS microscopy

  橋本 守, Harsono Cahyadi, 南川 丈夫, 新岡 宏彦, 荒木 勉

  OPTRONICS オプトロニクス社 33 (389) 74 - 78 0286-9659 2014/05 [Not refereed][Invited]
  
• 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 日本工業出版 25 (3) 16 - 20 0917-026X 2014/03 [Not refereed][Invited]
  
• Accumulation of advanced glycation end-products in human dentine

  Jiro Miura, Kantaro Nishikawa, Mizuho Kubo, Shuichiro Fukushima, Mamoru Hashimoto, Fumio Takeshige, Tsutomu Araki

  ARCHIVES OF ORAL BIOLOGY 59 (2) 119 - 124 0003-9969 2014/02 [Refereed][Not invited]
   

  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.
• Observation, Visualization and Inspection of Biological Specimen with Light

  ARAKI Tsutomu, HASHIMOTO Mamoru, FUKUSHIMA Shuichiro

  Systems, control and information システム制御情報学会 58 (10) 396 - 403 0916-1600 2014
• Fourth-order coherent Raman microspectroscopy for detection of material symmetry

  Mamoru Hashimoto, Hiroto Kanoh, Hirohiko Niioka, Tsutomu Araki

  Progress in Biomedical Optics and Imaging - Proceedings of SPIE 8948 1605-7422 2014 [Refereed][Not invited]
   

  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.
• A Stimulated Raman Scattering Imager Using High-Speed Lateral Electric Field Modulator and Lock-in Pixels Amplifiers

  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  0277-786X 2014 [Refereed][Not invited]
   

  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.
• Fourth-order coherent Raman microspectroscopy for detection of material symmetry

  Mamoru Hashimoto, Hiroto Kanoh, Hirohika Niioka, Tsutomu Araki

  MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES XIV 8948 894817  0277-786X 2014 [Refereed][Not invited]
   

  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.
• High-resolution microscopy for biological specimens via cathodoluminescence of Eu- and Zn-doped Y2O3 nanophosphors

  Taichi Furukawa, Hirohiko Niioka, Masayoshi Ichimiya, Tomohiro Nagata, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto

  Optics Express 21 (22) 25655 - 25663 1094-4087 2013/11/04 [Refereed][Not invited]
   

  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.
• Fast spectral coherent anti-Stokes Raman scattering microscopy with high-speed tunable picosecond laser

  Harsono Cahyadi, Junichi Iwatsuka, Takeo Minamikawa, Hirohiko Niioka, Tsutomu Araki, Mamoru Hashimoto

  JOURNAL OF BIOMEDICAL OPTICS 18 (9) 096009  1083-3668 2013/09 [Refereed][Not invited]
   

  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.
• Molecular Orientation Imaging of Liquid Crystals by Tunable-Polarization-Mode Coherent Anti-Stokes Raman Scattering Microscopy

  Takeo Minamikawa, Tatsuro Takagi, Hirohiko Niioka, Makoto Kurihara, Nobuyuki Hashimoto, Tsutomu Araki, Mamoru Hashimoto

  APPLIED PHYSICS EXPRESS 6 (7) 072401  1882-0778 2013/07 [Refereed][Not invited]
   

  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
• SHG顕微断層偏光イメージング : SHGによるコラーゲンの可視化 (特集 非線形光学効果を利用した生体イメージング)

  橋本 守, 福島 修一郎, 荒木 勉

  光アライアンス 日本工業出版 24 (4) 21 - 29 0917-026X 2013/04
• Application of Second-harmonic-generation Imaging to Tissue Characterization

  福島 修一郎, 橋本 守, 荒木 勉

  京都府立医科大学雑誌 京都府立医科大学 122 (4) 189 - 198 0023-6012 2013/04
• Coherent Anti-Stokes Raman Scattering Microscopy for High Speed Non-Staining Biomolecular Imaging

  Mamoru Hashimoto, Takeo Minamikawa, Tsutomu Araki

  CURRENT PHARMACEUTICAL BIOTECHNOLOGY 14 (2) 150 - 158 1389-2010 2013/02 [Refereed][Invited]
   

  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.
• Frequency-Swept Asynchronous-Optical-Sampling Terahertz Time-Domain Spectroscopy

  Yasui T, Iyonaga Y, Hsieh Y. D, Inaba H, Minoshima K, Yokoyama M, Araki T, Hashimoto M, Ieee

  2013 Conference on Lasers and Electro-Optics 2013 [Refereed][Not invited]
  
• A Time-Resolved CMOS Image Sensor With Draining-Only Modulation Pixels for Fluorescence Lifetime Imaging

  Zhuo Li, Shoji Kawahito, Keita Yasutomi, Keiichiro Kagawa, Juichiro Ukon, Mamoru Hashimoto, Hirohiko Niioka

  IEEE TRANSACTIONS ON ELECTRON DEVICES 59 (10) 2715 - 2722 0018-9383 2012/10 [Refereed][Not invited]
   

  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.
• CARS Microscopy: Implementation of nonlinear vibrational spectroscopy for far-field and near-field imaging

  Mamoru Hashimoto, Taro Ichimura, Katsumasa Fujita

  Springer Series in Optical Sciences 168 317 - 346 0342-4111 2012 [Refereed][Not invited]
   

  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.
• High-speed spectral tuning CARS microscopy using AOTF laser

  Mamoru Hashimoto, Junichi Iwatsuka, Hirohiko Niioka, Tsutomu Araki

  Progress in Biomedical Optics and Imaging - Proceedings of SPIE 8226 1605-7422 2012 [Refereed][Not invited]
   

  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).
• Multicolor Cathodoluminescence Microscopy for Biological Imaging with Nanophosphors

  Hirohiko Niioka, Taichi Furukawa, Masayoshi Ichimiya, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto

  APPLIED PHYSICS EXPRESS 4 (11) 1882-0778 2011/11 [Refereed][Not invited]
   

  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
• Real-time imaging of laser-induced membrane disruption of a living cell observed with multifocus coherent anti-Stokes Raman scattering microscopy

  Takeo Minamikawa, Hirohiko Niioka, Tsutomu Araki, Mamoru Hashimoto

  JOURNAL OF BIOMEDICAL OPTICS 16 (2) 021111  1083-3668 2011/02 [Refereed][Not invited]
   

  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]
• Development of polarization-mode controllable CARS microscope

  Mamoru Hashimoto, Tatsuro Takagi, Takeo Minamikawa, Hirohiko Niioka, Tsutomu Araki

  Progress in Biomedical Optics and Imaging - Proceedings of SPIE 7903 1605-7422 2011 [Refereed][Not invited]
   

  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.
• Photo-induced cell damage analysis for multi-focus CARS microscopy

  Takeo Minamikawa, Yoshinori Murakami, Naokazu Matsumura, Hirohiko Niioka, Shuichiro Fukushima, Tsutomu Araki, Mamoru Hashimoto

  Progress in Biomedical Optics and Imaging - Proceedings of SPIE 7903 1605-7422 2011 [Refereed][Not invited]
   

  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.
• Microscopic measurement of strain distribution on MEMS device using three dimensional orientation microscope

  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 1084-6999 2011 [Refereed][Not invited]
   

  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.
• Photo-induced cell damage analysis for multi-focus CARS microscopy

  Takeo Minamikawa, Yoshinori Murakami, Naokazu Matsumura, Hirohiko Niioka, Shuichiro Fukushima, Tsutomu Araki, Mamoru Hashimoto

  MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES XI 7903 79032H  0277-786X 2011 [Refereed][Not invited]
   

  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.
• Development of polarization-mode controllable CARS microscope

  Mamoru Hashimoto, Tatsuro Takagi, Takeo Minamikawa, Hirohiko Niioka, Tsutomu Araki

  MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES XI 7903 79031E  0277-786X 2011 [Refereed][Not invited]
   

  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.
• Development of strain visualization system for microstructures using single fluorescent molecule tracking on three dimensional orientation 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  0277-786X 2011 [Refereed][Not invited]
   

  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.
• Dedvelopement of Real-Time CARS Microscope and Application to Living Cell Measurement

  HASHIMOTO Mamoru, MINAMIKAWA Takeo, NIIOKA Hirohiko, ARAKI Tsutomu

  Nippon Laser Igakkaishi 特定非営利活動法人 日本レーザー医学会 30 (4) 421 - 426 0288-6200 2010/01/30 [Not refereed][Not invited]
   

  Raman microscopy visualizes molecular spices without any staining, because molecular vibrations that all molecules have are sensitive to molecular spices. We developed a real-time CARS (coherent anti-Stokes Raman scattering, which is one of nonlinear Raman scattering) microscopy system. We demonstrate three dimensional lipid distribution in a cell, laser-induced disruption of a lipid rich organelle, and plasma membrane repairing of disrupted plasma membrane.
• T0302-2-3 Noncontact and nondestructive measurement of strain using three-dimensional molecular orientation measurement under microscope

  YOSHIKI Keisuke, Masuda Kyosuke, HASHIMOTO Mamoru, HASHIMOTO Nobuyuki, KURIHARA Makoto, Namazu Takahiro, INOUE Shozo

  The proceedings of the JSME annual meeting 一般社団法人日本機械学会 2010 257 - 258 2010 

  We developed a system to analyze microscopic deformation, which can visualize inhomogeneous deformation in MEMS(Micro Electro Mechanical Systems) devices by using single-molecule markers. As MEMS devices become smaller, their mechanical properties seem to become inhomogeneous. This causes an undesirable distribution of deformation and stress, and decreases the reliability of the MEMS devices. We distributed fluorescent single molecules discretely on the microstructure of a MEMS device, and traced their relative displacements and rotations by observation under a three-dimensional orientation microscope, which provided information about normal, torsional, and bending displacements.
• High-speed CARS spectral imaging using acousto optic tunable filter

  Mamoru Hashimoto, Takeo Minamikawa, Tsutomu Araki

  Progress in Biomedical Optics and Imaging - Proceedings of SPIE 7569 1605-7422 2010 [Refereed][Not invited]
   

  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.
• Real-time molecular imaging of organelles in living cell by multifocus excitation CARS microscope

  Takeo Minamikawa, Tsutomu Araki, Mamoru Hashimoto

  Progress in Biomedical Optics and Imaging - Proceedings of SPIE 7569 1605-7422 2010 [Refereed][Not invited]
   

  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.
• SHG Microscopy Excitation with Radially Polarized Beam

  Hirohiko NIIOKA, Koichiro ASHIDA, Keisuke YOSHIKI, Tsutomu ARAKI, Mamoru HASHIMOTO

  Japanese journal of optics 応用物理学会分科会日本光学会 39 (8) 401-403 - 403 0389-6625 2010 [Refereed][Invited]
  
• High-speed CARS spectral imaging using acousto optic tunable filter

  Mamoru Hashimoto, Takeo Minamikawa, Tsutomu Araki

  MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES X 7569 75690Q  0277-786X 2010 [Refereed][Not invited]
   

  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.
• Real-time molecular imaging using multi-focus excitation CARS microscope

  南川 丈夫, 荒木 勉, 橋本 守

  Optronics オプトロニクス社 28 (8) 103 - 107 0286-9659 2009/08 [Not refereed][Not invited]
  
• Multi-focus excitation coherent anti-Stokes Raman scattering (CARS) microscopy and its applications for real-time imaging

  Takeo Minamikawa, Mamoru Hashimoto, Katsumasa Fujita, Satoshi Kawata, Tsutomu Araki

  OPTICS EXPRESS 17 (12) 9526 - 9536 1094-4087 2009/06 [Refereed][Not invited]
   

  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
• Enhancement of second-harmonic generation from self-assembled monolayers on gold by excitation with a radially polarized beam

  Mamoru Hashimoto, Koichiro Ashida, Keisuke Yoshiki, Tsutomu Araki

  OPTICS LETTERS 34 (9) 1423 - 1425 0146-9592 2009/05 [Refereed][Not invited]
   

  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
• Multi-focus CARS microscopy using microlens array scanner for realtime molecular spectral-imaging

  Mamoru Hashimoto, Takeo Minamikawa, Tsutomu Araki, Katsumasa Fujita, Satoshi Kawata

  Progress in Biomedical Optics and Imaging - Proceedings of SPIE 7183 1605-7422 2009 [Refereed][Not invited]
   

  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.
• Lipids distribution imaging of lipid vesicles by multi-focus excitation CARS microscope

  Takeo Minamikawa, Tsutomu Araki, Mamoru Hashimoto

  Progress in Biomedical Optics and Imaging - Proceedings of SPIE 7183 1605-7422 2009 [Refereed][Not invited]
   

  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.
• Multifocus CARS microscopy for realtime vibrational imaging

  Mamoru Hashimoto, Takeo Minamikawa, Hirohiko Niioka, Tsutomu Araki

  Proceedings of SPIE - The International Society for Optical Engineering 7507 75070H  0277-786X 2009 [Refereed][Not invited]
   

  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.
• Multi-focus CARS microscopy using microlens array scanner for realtime molecular spectral-imaging

  Mamoru Hashimoto, Takeo Minamikawa, Tsutomu Araki, Katsumasa Fujita, Satoshi Kawata

  MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES IX 7183 0277-786X 2009 [Not refereed][Not invited]
   

  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).
• Lipids distribution imaging of lipid vesicles by by multi-focus excitation CARS microscope

  Takeo Minamikawa, Tsutomu Araki, Mamoru Hashimoto

  MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES IX 7183 718328  0277-786X 2009 [Refereed][Not invited]
   

  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.
• 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 日本工業出版 Vol. 20, No. 3 (3) 24 - 28 0917-026X 2009 [Not refereed][Not invited]
  
• 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 日本工業出版 20 (4) 21 - 25 0917-026X 2009 [Not refereed][Not invited]
  
• 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 0386-2178 2009 [Not refereed][Not invited]
  
• High Resolution Imaging of Cell Organelle Using Compact Polarization Mode Converter

  Keisuke YOSHIKI, Tomomasa AIKAWA, Mamoru HASHIMOTO, Makoto KURIHARA, Nobuyuki HASHIMOTO, Tsutomu ARAKIÐ

  BME 社団法人日本生体医工学会 46 (6) 698 - 702 1347-443X 2008 [Refereed][Not invited]
   

  The shape of focal spot is affected by the distribution of polarization and phase on the cross-section of the incident beam. We demonstrated the spatial resolution enhancement effect using the difference of the focal spots between a linearly polarized beam and an azimuthally polarized beam. The azimuthally polarized beam has the vortex of polarization on the cross-section of the beam. Since the vortex of the azimuthally polarized beam forms a doughnut shaped focal spot, the spatial resolution is expected to increase with the difference image between linearly polarized beam excitation/detection and azimuthally polarized beam excitation/detection. We applied an eight segmented polarization mode converter, which was developed by ourselves and could switch those polarization modes of the excitation beam electrically, to a commercial confocal microscope. We derived the suitable weight of subtraction theoretically, and demonstrated the spatial resolution enhancement effect by observing the stained cell cytoskeleton.
• Real-time terahertz color scanner for moving objects

  Takeshi Yasui, Ken-ichi Sawanaka, Atsushi Ihara, Emmanuel Abraham, Mamoru Hashimoto, Tsutomu Araki

  OPTICS EXPRESS 16 (2) 1208 - 1221 1094-4087 2008/01 [Refereed][Not invited]
   

  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.
• Second-harmonic-generation microscope using eight-segment polarization-mode converter to observe three-dimensional molecular orientation: publisher's note (vol 32, pg 1680, 2007)

  Keisuke Yoshiki, Ryosuke Kanamaru, Mamoru Hashimoto, Nobuyuki Hashimoto, Tsutomu Araki

  OPTICS LETTERS 32 (16) 2465 - 2465 0146-9592 2007/08 [Refereed][Not invited]
  
• Second-harmonic-generation microscope using eight-segment polarization-mode converter to observe three-dimensional molecular orientation

  Keisuke Yoshiki, Kanamaru Ryosuke, Mamoru Hashimoto, Tsutomu Araki, Nobuyuki Hashimoto

  OPTICS LETTERS 32 (12) 1680 - 1682 0146-9592 2007/06 [Refereed][Not invited]
   

  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
• A compact polarization converter to observe molecular orientation

  Mamoru Hashimoto, Keisuke Yoshiki, Ryosuke Kanamaru, Nobuyuki Hashimoto, Tsutomu Araki

  THREE-DIMENSIONAL AND MULTIDIMENSIONAL MICROSCOPY: IMAGE ACQUISITION AND PROCESSING XIV 6443 64430L  0277-786X 2007 [Refereed][Not invited]
   

  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.
• Second Harmonic Generation Microscopy with Polarization Pattern Control of Exaction Beam Using Liquid-Crystal Spatial-Light-Modulator

  Mamoru HASHIMOTO

  光学 36 (3) 143 - 148 2007 [Not refereed][Not invited]
  
• Jitter reduction of two synchronized picosecond mode-locked lasers using balanced cross-correlator with two-photon detectors

  Takeo Minamikawa, Naoki Tanimoto, Mamoru Hashimoto, Tsutomu Araki, Minoru Kobayashi, Katsumasa Fujita, Satoshi Kawata

  APPLIED PHYSICS LETTERS 89 (19) 191101  0003-6951 2006/11 [Refereed][Not invited]
   

  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.
• 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 高温学会レーザ加工学会 13 (2) 140 - 143 2006/04 [Not refereed][Not invited]
  
• SHG microscopy excited by polarization controlled beam for three-dimensional molecular orientation measurement

  K. Yoshiki, M. Hashimoto, T. Araki

  LASER BEAM SHAPING VII 6290 62900F  0277-786X 2006 [Refereed][Not invited]
   

  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.
• Finding of optimal calcium ion probes for fluorescence lifetime measurement

  K Yoshiki, H Azuma, K Yoshioka, M Hashimoto, T Araki

  OPTICAL REVIEW 12 (5) 415 - 419 1340-6000 2005/09 [Refereed][Not invited]
   

  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.
• 「平成17年度春季講演会・シンポジウム」報告

  橋本 守

  分光研究 = Journal of the spectroscopical research of Japan The Spectroscopical Society of Japan 54 (3) 180 - 180 0038-7002 2005/06/15
• Automatic pulse duration control of picose cond laser using two-photon absorption detector

  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 0021-4922 2005/06 [Refereed][Not invited]
   

  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.
• Multi-focus CARS microscopy using automatic pulse duration control system

  M Hashimoto, T Asada, T Araki, S Kawata

  2005 PACIFIC RIM CONFERENCE ON LASERS AND ELECTRO-OPTICS 660 - 661 2005 [Refereed][Not invited]
   

  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.
• Tip-enhanced near-field CARS microscopy for molecular nano-imaging

  S Kawata, T Ichimura, N Hayazawa, M Hashimoto, Y Inouye

  MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES V 5700 52 - 59 0277-786X 2005 [Refereed][Not invited]
   

  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.
• 10 fs レーザー用自己相関計の製作

  橋本 守, 浅田崇裕, 荒木 勉

  分光研究 The Spectroscopical Society of Japan 54 (1) 32 - 34 0038-7002 2005 [Refereed][Not invited]
  
• Second-harmonic-generation microscopy using excitation beam with controlled polarization pattern to determine three-dimensional molecular orientation

  K Yoshiki, M Hashimoto, T Araki

  JAPANESE JOURNAL OF APPLIED PHYSICS PART 2-LETTERS & EXPRESS LETTERS 44 (33-36) L1066 - L1068 0021-4922 2005 [Refereed][Not invited]
   

  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.
• Proposition of single molecular orientation determination using polarization controlled beam by liquid crystal spatial light modulators

  M Hashimoto, K Yamada, T Araki

  OPTICAL REVIEW 12 (1) 37 - 41 1340-6000 2005/01 [Refereed][Not invited]
   

  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.
• Tip-enhanced near-field CARS microscopy

  S Kawata, T Ichimura, N Hayazawa, Y Inouye, M Hashimoto

  JOURNAL OF NONLINEAR OPTICAL PHYSICS & MATERIALS 13 (3-4) 593 - 599 0218-8635 2004/12 [Not refereed][Invited]
   

  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.
• Tip-enhanced coherent anti-Stokes Raman scattering for vibrational nanoimaging

  T Ichimura, N Hayazawa, M Hashimoto, Y Inouye, S Kawata

  PHYSICAL REVIEW LETTERS 92 (22) 220801  0031-9007 2004/06 [Refereed][Not invited]
   

  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.
• Coherent anti-stokes Raman nano-imaging with metal-tip field enhancement

  N Hayazawa, T Ichimura, M Hashimoto, Y Inouye, S Kawata

  ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY 227 U280 - U280 0065-7727 2004/03 [Not refereed][Invited]
  
• Amplification of coherent anti-Stokes Raman scattering by a metallic nanostructure for a high resolution vibration microscopy

  N Hayazawa, T Ichimura, M Hashimoto, Y Inouye, S Kawata

  JOURNAL OF APPLIED PHYSICS 95 (5) 2676 - 2681 0021-8979 2004/03 [Refereed][Not invited]
   

  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.
• Application of tip-enhanced microscopy for nonlinear Raman spectroscopy

  T Ichimura, N Hayazawa, M Hashimoto, Y Inouye, S Kawata

  APPLIED PHYSICS LETTERS 84 (10) 1768 - 1770 0003-6951 2004/03 [Refereed][Not invited]
   

  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.
• Coherent anti-Stokes Raman spectroscopy for nano-imaging with a metallic near-field probe

  S Kawata, T Ichimura, N Hayazawa, M Hashimoto, Y Inouye

  NONLINEAR OPTICAL TRANSMISSION AND MULTIPHOTON PROCESSES IN ORGANICS II 5516 1 - 8 0277-786X 2004 [Refereed][Not invited]
   

  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.
• Tip-enhanced NSOM

  S Kawata, N Hayaazawa, T Yano, H Watanabe, T Ichimura, M Hashimoto, Y Inouye

  NANOBIOPHOTONICS AND BIOMEDICAL APPLICATIONS 5331 1 - 12 0277-786X 2004 [Refereed][Not invited]
   

  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.
• Local enhancement of coherent anti-Stokes Raman scattering by isolated gold nanoparticles

  T Ichimura, N Hayazawa, M Hashimoto, Y Inouye, S Kawata

  JOURNAL OF RAMAN SPECTROSCOPY 34 (9) 651 - 654 0377-0486 2003/09 [Refereed][Not invited]
   

  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.
• Development of Coherent Anti - Stokes Raman Scattering Microscopy

  HASHIMOTO Mamoru

  rle The Laser Society of Japan 31 (6) 375 - 379 0387-0200 2003/06/15 

  New multiphoton microscopy using CARS (Coherent Anti-Stokes Raman Scattering) spectroscopy is described. In CARS microscopy, the image of molecular vibration that is sensitive to the molecular spices and conformation is obtainable without staining with three-dimensional resolution of sub-micrometer. In this article, optical property of CARS microscopy by the diffraction theory and the developed system using a picosecond tunable laser are described. The multi-focus excitation using a rotating-microlens array enables high speed spectral imaging.
• High Precision, White Nanosecond Xe Pulsed Light Source for High-Speed Stroboscopic Measurement

  ITAMI Shin, HASHIMOTO Mamoru, ARAKI Tsutomu

  Transactions of the Japan Society of Mechanical Engineers. Series C. 一般社団法人日本機械学会 69 (684) 210 - 215 0387-5024 2003 [Refereed][Not invited]
   

  We have developed a thyratron-gated, white nanosecond pulsed light source for high-speed stroboscopic measurement using a commercially available Xe shorr-arc lamp. To reduce time jitter, a grounded grid thyratron and an avalanche transistor are used for this light source. The grounded grid thyratron plays the role as switching component for high voltage and large current, and the avalanche transistor plays the role as main parts of fast trigger pulse generator. Time jitter was determined quantitatively using time to amplitude converter (TAC). The minimum value and maximum value of time jitter were 1.44 ns (FWHM : full width at half-maximum) and 4.5 ns (FWHM), respectively. As time jitter of the previous pulsed light source using grounded cathode thyratron was 8 ns, sufficient results were obtained. Intense light pulses as large as 85 W (peak value) of 15.3 ns duration (FWHM) was obtained from the proposed Xe lamp system. The reduction of the time jitter is suitable for an excitation light source in time-resolved spectroscopy and also for a stroboscopic illumination in a high-speed fluid experiment.
• Coherent Anti-Stokes Raman Scattering Microscopy

  HASHIMOTO Mamoru

  Japanese journal of optics 応用物理学会分科会日本光学会 31 (6) 496 - 498 0389-6625 2002/06/10
• Coherent anti-Stokes Raman scattering microscopy

  M Hashimoto

  ACTA HISTOCHEMICA ET CYTOCHEMICA 35 (2) 83 - 86 0044-5991 2002 [Refereed][Invited]
   

  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.
• Visual demonstration of calcium accumulation in human arteries of upper and lower limbs

  Y Tohno, S Tohno, Y Tateyama, Y Kida, T Yasui, M Hashimoto, T Araki

  BIOLOGICAL TRACE ELEMENT RESEARCH 81 (2) 115 - 125 0163-4984 2001/08 [Not refereed][Invited]
   

  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.
• Three-dimensional transfer functions of coherent anti-Stokes Raman scattering microscopy

  M Hashimoto, T Araki

  JOURNAL OF THE OPTICAL SOCIETY OF AMERICA A-OPTICS IMAGE SCIENCE AND VISION 18 (4) 771 - 776 0740-3232 2001/04 [Refereed][Not invited]
   

  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.
• Three-dimensional transfer functions of coherent anti-stokes raman scattering microscopy

  Mamoru Hashimoto, Tsutomu Araki

  Journal of the Optical Society of America A: Optics and Image Science, and Vision 18 (4) 771 - 776 1520-8532 2001 [Refereed][Not invited]
   

  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.
• Molecular vibration imaging in the fingerprint region by use of coherent anti-Stokes Raman scattering microscopy with a collinear configuration

  M Hashimoto, T Araki, S Kawata

  OPTICS LETTERS 25 (24) 1768 - 1770 0146-9592 2000/12 [Refereed][Not invited]
   

  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.
• Correlations of calcium accumulations in arteries, veins, cartilages, ligaments, and bones in single humans

  Y Tateyama, Y Takano, Y Tohno, Y Moriwake, S Tohno, M Hashimoto, T Araki

  BIOLOGICAL TRACE ELEMENT RESEARCH 74 (3) 211 - 221 0163-4984 2000/06 [Refereed][Not invited]
   

  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.
• Coherent Anti-Stokes Raman Scattering Microscopy

  HASHIMOTO mamoru

  Bunko Kenkyu The Spectroscopical Society of Japan 49 (2) 51 - 61 0038-7002 2000 [Refereed][Not invited]
  
• Coherent anti-stokes Raman scattering microscope

  M Hashimoto, T Araki

  18TH CONGRESS OF THE INTERNATIONAL COMMISSION FOR OPTICS: OPTICS FOR THE NEXT MILLENNIUM, TECHNICAL DIGEST 3749 496 - 497 0277-786X 1999 [Refereed][Not invited]
   

  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.
• Real-time pursuit of crystal growth by millisecond time-resolved multichannel Fourier transform infrared spectroscopy

  Mamoru Hashimoto, Hiro-O. Hamaguchi

  Applied Spectroscopy 52 (2) 222 - 225 0003-7028 1998 [Refereed][Not invited]
   

  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.
• Time-resolved infrared study of ground-state phototautomer formed in the excited-state proton transfer of 7-hydroxyquinoline in methanol

  K Tokumura, M Natsume, T Nakagawa, M Hashimoto, T Yuzawa, H Hamaguchi, M Itoh

  CHEMICAL PHYSICS LETTERS 271 (4-6) 320 - 326 0009-2614 1997/06 [Refereed][Not invited]
   

  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.
• Temporal emission characteristics of light emitting-diodes for high-speed pulsed current

  Y. Fujisawa, M. Hashimoto, T. Araki

  Journal of the Illuminating Engineering Institute of JAPAN 81 (8A) 656 - 663 1997/05 [Refereed][Not invited]
  
• An ultraviolet nanosecond light pulse generator using a light emitting diode for test of photodetectors

  Tsutomu Araki, Yasumitsu Fujisawa, Mamoru Hashimoto

  Review of Scientific Instruments 68 (3) 1365 - 1368 0034-6748 1997 [Refereed][Not invited]
   

  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.
• Construction of a multichannel Fourier transform infrared spectrometer for single-event time-resolved spectroscopy

  M Hashimoto, HO Hamaguchi

  APPLIED SPECTROSCOPY 50 (8) 1030 - 1033 0003-7028 1996/08 [Refereed][Not invited]
   

  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.
• Nanosecond time resolved infrared spectroscopy and its application to photomolecular science.

  M. Hashimoto, T. Yuzawa, H. Hamaguchi

  Jacso Report 37 (3) 27 - 33 0916-3492 1995/06 [Refereed][Invited]
  
• Structure of the Twisted-Intramolecular-Charge-Transfer Excited Singlet and Triplet States of 4-(Dimethylamino)benzonitrile As Studied by Nanosecond Time-Resolved Infrared Spectroscopy

  Mamoru Hashimoto, Hiro-o Hamaguchi

  The Journal of Physical Chemistry 99 (20) 7875 - 7877 0022-3654 1995/05 [Refereed][Not invited]
  
• Signal to noise ratio of multi-channel Fourier-transform spectroscopy

  M. Hashimoto, S. Kawata

  J. Spectrosc. Soc. Jpn. The Spectroscopical Society of Japan 41 (5) 317 - 326 0038-7002 1992/08 [Refereed][Not invited]
   

  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.
• Multichannel fourier-transform infrared spectrometer

  Mamoru Hashimoto, Satoshi Kawata

  Applied Optics 31 (28) 6096 - 6101 2155-3165 1992 [Refereed][Not invited]
   

  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.
• MULTICHANNEL FT-IR SPECTROMETER WITH A 4096-ELEMENT INFRARED CCD

  Satoshi Kawata, Mamoru Hashimoto

  ANALYTICAL SCIENCES 7 575 - 576 0910-6340 1991 [Refereed][Not invited]
   

  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)

Books etc

• CARS Microscopy: Implementation of nonlinear vibrational spectroscopy for far-field and near-field imaging

  Mamoru Hashimoto, Taro Ichimura, Katsumasa Fujita 
  2012 

  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.
• 顕微分光法 ナノ・マイクロの世界を見る分光法

  (Contributor赤外・ラマン顕微分光法)
  講談社サイエンティフィク 2009
• バイオイメージングがわかる

  橋本守, 福島修一郎, 荒木勉 (ContributorLIM蛍光寿命イメージング)
  2005
• Biological Imaging and Sensing

  S. Kawata, O. Nakamura, T. Kaneko, M. Hashimoto, K. Goto, N. I. Smith, T. Sugiura, I. Fujimasa, H. Matsumoto (ContributorBiological imaging and sensing from basic techniques to clinical application)
  Springer 2004 1-68
• Handbook of Vibrational Spectroscopy

  Mamoru Hashimoto, Teturo Yuzawa, Chihiro Kato, Koichi Iwata, Hiro‐o Hamaguchi (ContributorFast time-resolved mid-infrared spectroscopy using grating spectrometers)
  John Wiley & Sons, Ltd 2001 666-676

Conference Activities & Talks

• CMOSイメージセンサーを用いたマルチプレックス誘導ラマン散乱分光法”, "新しいイメージングを実現する最先端CMOSイメージセンサ

  De Xing Lioe, Shukri Bin Korakkottil, Kunhi Mohd, 本間 宗一郎, 大和 尚記, 安富 啓太, 香川 景一郎, 橋本 守, 川人 祥二

  Optics & Photonics Japan 2023  2023/11
• Development of Lissajous scanning fluorescence endomicroscope using polarization-maintaining fiber

  N. Yamato, M. Hashimoto

  OSJ-JSAP Joint Symposia on Optics, Optics & Photonics Japan 2023  2023/11
• Imaging of trans lipid-mobilization by coherent Raman microspectroscopy  [Not invited]

  S. Homma, N. Yamato, M. Hashimoto

  OSJ-JSAP Joint Symposia on Optics, Optics & Photonics Japan 2023  2023/11
• Label-free imaging by coherent Raman scattering endoscopy  [Invited]

  M. Hashimoto

  2023 International Workshop on the New Frontiers in Convergence Science and Technology of Hokkaido University - Seoul National Universty 26th Joint Symposium  2023/11
• 第二高調波発生関節鏡の開発とビーム走査手法の検討  [Invited]

  橋本守

  第21回医用分光学研究会  2023/10
• 光ファイバーレーザーを用いた無染色コラーゲン可視化間接鏡の開発

  橋本 守, 大和 尚紀, 松田 陸

  第62回日本生体医工学会北海道支部大会  2023/10
• Multiplex coherent anti-Stokes Raman 散乱分光顕微鏡を用いた脂肪様細胞のトランス脂肪動員観察  [Not invited]

  本間 宗一郎, 大和 尚記, 橋本 守

  第84回応用物理学会秋季学術講演会  2023/09
• Lipid metabolism imaging by elliptical spots scanning multiplex coherent anti-stokes raman scattering spectromicroscopy  [Invited]

  M. Hashimoto

  8th Asian Spectroscopy Conference 2023  2023/09
• Examination of polarization compensation method using two electro-optic modulators  [Not invited]

  R. Ikazaki, Y. Kawasaki, N. Yamato, M. Hashimoto

  Abstracts of the 58th Hokkaido Branch of the Japan Society of Applied Physics / the 19th Hokkaido Branch of the Optical Society of Japan  2023/01
• 楕円スポット走査マルチプレックスCARS 顕微分光イメージング  [Invited]

  橋本守

  第19 回医用分光学研究会  2021/11
• Ybファイバーレーザーを用いた第二高調波発生関節鏡の開発

  松田陸, 橋本守

  Optics & Photonics Japan 2021  2021/10
• 楕円スポット型コヒーレント反ストークスラマン散乱分光顕微鏡のCARS光強度増強効果の実験的検証  [Not invited]

  本間宗一郎, 橋本守

  第82回応用物理学会秋季学術講演会  2021/09
• Real-time nerve extraction using coherent raman scattering rigid endoscope and deep learning  [Not invited]

  N. Yamato, M. Matsuya, H. Niioka, J. Miyake, M. Hashimoto

  The 60 th Annual Conference of Japan Society for Medical and Biological Engineering, and The Anuual Meeting of Japan Biomagnetism and Bioelectromagnetics Society  2021/06
• Evaluation of controlled release from doxorubicin-loaded nanoparticles under ultrasound irradiation

  M. Shinzato, Y. Kato, M. Hashimoto, N. Kudo

  The 60 th Annual Conference of Japan Society for Medical and Biological Engineering, and The Anuual Meeting of Japan Biomagnetism and Bioelectromagnetics Society  2021/06
• CARS硬性内視鏡の神経イメージングと深層学習による神経抽出ー実時間イメージングに向けた短時間露光神経画像の画像解析ー  [Not invited]

  大和尚記, 松谷真奈, 新岡宏彦, 三宅淳, 橋本守

  第68回応用物理学会春季学術講演会  2021/03
• 非線形ラマン散乱を用いた顕微内視鏡の開発  [Invited]

  橋本守

  4大学医工連携オンラインセミナー ～光×超音波×近赤外蛍光による医工連携イメージングと健康長寿への道～（onLine）  2021/02
• 楕円スポットを用いたコヒーレント反ストークスラマン散乱分光顕微鏡による信号増強効果の実験的検証

  本間 宗一郎, 阿部 隆爾, 橋本 守

  第 56 回 応用物理学会北海道支部／第 17 回 日本光学会北海道支部 合同学術講演会  2021/01
• 光ファイバーバンドルを用いた非線形ラマン散乱顕微内視鏡  [Invited]

  橋本守, 小川拓希

  第18回医用分光学研究会  2020/11
• 非線形ラマン散乱内視鏡に向けた光ファイバーバンドル内での四光波混合発生の低減法  [Not invited]

  小川拓希, 橋本守

  Optics & Photonics Japan 2020  2020/11
• 光ファイバーバンドルを用いた非線形ラマン散乱内視鏡の開発  [Invited]

  橋本守

  北海道大学 新技術説明会  2020/11
• 神経を可視化するコヒーレントアンチストークスラマン散乱硬性内視鏡の可搬化のためのファイバーレーザー光源の開発

  松田陸, 大和尚記, 橋本守

  第59回日本生体医工学会北海道支部大会  2020/10
• 非線形ラマン散乱硬性内視鏡と深層学習による無染色な末梢神経イメージングの検討  [Invited]

  大和尚記, 松谷真奈, 新岡宏彦, 三宅淳, 橋本守

  日本蛍光ガイド手術研究会第3回学術集会  2020/10
• 非線形ラマン散乱硬性内視鏡と深層学習による神経イメージング装置の開発  [Not invited]

  大和尚記, 松谷真奈, 新岡宏彦, 三宅淳, 橋本守

  レーザー学会第547回研究会  2020/10
• High-speed hyperspectral imaging of living cells with beam scanning and slit acquisition polarization coherent {anti-Stokes} {Raman} scattering microspectroscopy

  Ryuji Abe, Mamoru Hashimoto

  The 81st JSAP Autumn Meeting  2020/09
• 非線形ラマンイメージングの高速化 –深層学習と楕円形スポット–  [Invited]

  橋本 守

  日本学術振興会 生体ひかりイメージング技術と応用第185委員会  2020/09
• ノイズ除去のアンサンブル学習による非線形ラマン硬性内視鏡神経イメージングの高速化

  大和尚記, 新岡宏彦, 橋本守

  第59回日本生体医工学会大会  2020/05
• High-speed hyperspectral imaging of living cells with slit-scanning multiplex coherent {anti-Stokes} {Raman} scattering microspectroscopy using elliptical focal spot

  Ryuji Abe, Kyotaro Horio, Shun Kizawa, Mamoru Hashimoto

  The 67th JSAP spring Meeting. Meeting was canceld (Abstract was published).  2020/03
• 非線形ラマン硬性鏡による神経イメージの人工知能解析  [Invited]

  橋本守

  レーザー学会学術講演会第40回年次大会  2020/01
• 二波長の後方散乱光を用いた無侵襲静脈血中脂質濃度推定法の提案  [Not invited]

  下川部一真, 橋本守, 加藤祐次

  レーザー学会学術講演会第40回年次大会  2020/01
• Comparison of deep learning models to improve imaging speed of coherent {Raman} scattering rigid endoscopy

  Naoki Yamato, Niioka Hirohiko, Jun Miyake, Mamoru Hashimoto

  The 55th JSAP Hokkaido Branch The 16th OSJ Hokkaido Branch Joint Meeting  2020/01
• 非線形ラマン散乱硬性内視鏡と深層学習による神経イメージング  [Not invited]

  大和尚記, 松谷真奈, 工藤信樹, 新岡宏彦, 三宅淳, 橋本守

  第32回日本内視鏡外科学会総会  2019/12
• Nerve imaging and segmentation used by coherent Raman endoscopy and deep learning  [Invited]

  M. Hashimoto, N. Yamato, M. Matsuya, H. Niioka, J. Miyake

  Biomedical Raman Imaging 2019  2019/11
• スリット走査マルチプレックスCARS顕微鏡の開発  [Invited]

  橋本守, 木澤駿

  第17回医用分光学研究会  2019/11
• Live-cell observation with slit-scanning multiplex {C},{A},{R},{S} microspectroscopy using an elliptical focal spot  [Not invited]

  Shun Kizawa, Mamoru Hashimoto

  The 80th JSAP Autumn Meeting 2019  2019/09
• 転移学習を用いた深層学習による非線形ラマン像からの神経セグメンテーション  [Not invited]

  松谷真奈, 大和尚記, 新岡彦, 工藤信樹, 三宅淳, 橋本守

  第80回応用物理学会秋季学術講演会  2019/09
• コヒーレント反ストークスラマン散乱硬性鏡の開発と神経検出への応用  [Invited]

  橋本守

  第3期 第5回レーザー学会「レーザーバイオ医療」技術専門委員会  2019/07
• Deep Learningと転移学習を用いたCARS硬性内視鏡イメージングの高速化  [Not invited]

  新岡 宏彦, 大和 尚記, 三宅 淳, 橋本 守

  レーザ顕微鏡研究会第44回講演会・シンポジウム  2019/07
• 非線形ラマン散乱硬性鏡による神経イメージングの転移学習を用いた高速化  [Not invited]

  大和尚記, 新岡宏彦, 橋本守

  第58回日本生体医工学会大会  2019/06
• Nerve segmentation from fluorescence images by using transferred deep learning

  Mana Matsuya, Naoki Yamato, Hirohiko Niioka, Mamoru Hashimoto

  Japanese Society of Molecular Imaging 2019  2019/05
• 楕円スポットを用いたスリット走査型マルチプレックスコヒーレントアンチストークスラマン散乱分光顕微鏡の開発  [Not invited]

  木澤駿, 橋本守

  第66回応用物理学会春季学術講演会  2019/03
• CARS 硬性鏡による神経イメージング  [Invited]

  橋本守

  第39回レーザー学会学術講演会 年次大会  2019/01
• 非線形ラマン散乱硬性鏡を用いた神経イメージング  [Invited]

  橋本守

  第57回日本生体医工学会大会  2018/06
• 非線形ラマンを用いたラベルフリージング  [Invited]

  橋本守

  一般社団法人レーザー学会学術講演会第３８回年次大会  2018/01
• Development of coherent anti-Stokes Raman scattering rigid endoscope for label-free nerve imaging during an operation

  Hirose Keigo, Aoki Takuya, Fukushima Shuichiro, Sakaguchi Yoshiyuki, Furukawa Taichi, Hashimoto Mamoru

  Transactions of Japanese Society for Medical and Biological Engineering  2016  Japanese Society for Medical and Biological Engineering
   

  We have developed a coherent anti-Stokes Raman scattering (CARS) rigid endoscope for label-free nerve imaging during an operation. The CARS rigid endoscope consists of a single-mode fiber which delivers two color lasers, a galvano scanning system, and homemade ϕ12 mm lens tubes which include two relay lenses and an objective lens. The label-free imaging of rat sciatic nerve is demonstrated. To attain CARS image with high- speed, correcting chromatic aberration is focused on. The split between the focal spots of two lasers passed through the lens tubes is 21 μm and a numerical simulation shows that the intensity of CARS signal by correcting the chromatic aberration is 2.3 times as strong as non-correcting. The rigid endoscope has been modified to suppress the chromatic aberration by using two single-mode fibers which deliver the two lasers individually. The experimental result shows the increase of CARS intensity at least 1.5 times.

• Label free bioimaging using nonlinear coherent Raman microscopy  [Not invited]

  Mamoru Hashimoto

  IEEE CPMT Symposium Japan 2015: Packaging is Everywhere, ICSJ 2015  2015/12  Institute of Electrical and Electronics Engineers Inc.
   

  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.
• Nonlinear Raman microscopy for biomedical applications(Workshop on microscopy, biology, medicine, and advanced CMOS imagers)

  Hashimoto Mamoru, Kawahito Shoji

  ITE Technical Report  2015/11  The Institute of Image Information and Television Engineers
   

  Nonlinear Raman imaging enables high-speed visualization of the biomolecules without staining. We have developed a multimodal microscopy and an endoscopy systems observing nonlinear Raman scattering. The developed systems are applied to the imaging of atherosclerosis legion. The performances of label-free imaging are demonstrated. Lock-in image sensor for parallel excitation/detection of stimulated Raman imaging is also demonstrated.
• Label free bioimaging using nonlinear coherent Raman microscopy  [Invited]

  M. Hashimoto

  2015 IEEE CPMT Symposium Japan (ICSJ)  2015/11
• Nonlinear Raman microscopy for biomedical applications  [Invited]

  M. Hashimoto, S. Kawahito

  2015/11
• PS2-3 Label free imaging of atherosclerotic lesions using stimulated Raman scattering, second harmonic generation, and two-photon fluorescence microscopy(PS2: Poster Short Presentation II,Poster Session)

  Aoki Takuya, Tao Tomoyo, Fukushima Syuichiro, Araki Tsutomu, Hashimoto Mamoru

  Proceedings of the ... Asian Pacific Conference on Biomechanics : emerging science and technology in biomechanics  2015/09  The Japan Society of Mechanical Engineers
  
• PS2-14 OBSERVATION OF CELLULLAR RESPONSE TO OXYGEN TENSION USING MICROFLUIDIC DEVICES(PS2: Poster Short Presentation II,Poster Session)

  Fujitaka Naoya, Fukushima Shuichiro, Hashimoto Mamoru, Funamoto Kenichi, Araki Tsutomu

  Proceedings of the ... Asian Pacific Conference on Biomechanics : emerging science and technology in biomechanics  2015/09  The Japan Society of Mechanical Engineers
  
• Plasmonic-nanoparticle-enhanced hyper-Raman spectroscopy  [Not invited]

  B. Ranjan, M. Hashimoto, Y. Saito, P. Verma

  2015/09
• Label free imaging of atherosclerotic lesions using stimulate Raman scattering, second harmonic generation, and two-photon fluorescence microscopy  [Not invited]

  T. Aoki, T. Tao, S. Fukushima, T. Araki, M. Hashimoto

  Abstract of the 8th Asian-Pacific Conference on Biomechanics  2015/09
• Discrete fourier transform infrared spectroscopy using precisely periodic pulse  [Not invited]

  Y.-D. Hsieh, S. Okubo, H. Inaba, M. Hashimoto, T. Yasui

  2015/05
• Imaging of atherosclerosis legion using multimodal nonlinear optical microscopy  [Invited]

  M. Hashimoto

  2015/05
• Biological high-resolution imaging in wet-condition using cathodoluminescence microscopy  [Not invited]

  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  2015/05
• 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  [Not invited]

  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

  2015/03
• Discrete Fourier Transform Infrared Spectroscopy Using Precisely Periodic Pulse  [Not invited]

  Y. -D. Hiseh, S. Okubo, H. Inaba, M. Hashimoto, T. Yasui

  2015 CONFERENCE ON LASERS AND ELECTRO-OPTICS (CLEO)  2015  IEEE
   

  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.
• Highly luminescent rare-earth doped Y2O3 ceramics in near-infrared region by codoping Li+ ions and its application for imaging  [Not invited]

  J. Yamasaki, S. Fukushima, H. Niioka, T. Araki, M. Hashimoto, J. Miyake

  2014/12
• Gadolinium oxide doped rare earth nanophosphors for trimodal imaging  [Not invited]

  D. T, K. Dung, S. Fukushima, H. Niioka, M. Ichimiya, M. Ashida, T. Araki, M. Hashimoto, J. Miyake

  2014/12
• Observation of organic nonlinear optical crystal by multiplex fourth order Raman microscope  [Not invited]

  C. Ninagawa, T. furukawa, H. Niioka, T. Araki, M. Hashimoto

  2014/12
• A low-noise stimulated Raman scattering CMOS imager using high-speed lock-in pixels(Poster session,2nd Asian Image Sensors and Imaging Systems Symposium)

  Lioe De Xing, Mars Kamel, Han Sang Man, Takasawa Taishi, Yasutomi Keita, Kagawa Keiichiro, Hashimoto Mamoru, Kawahito Shoji

  ITE Technical Report  2014/11  The Institute of Image Information and Television Engineers
   

  We demonstrate a low-noise stimulated Raman scattering (SRS) CMOS imager using high-speed lock-in pixels. To detect small SRS signal under an extremely large offset due to probing laser signal of which the ratio is 10^<-5>, a lock-in pixel using a high-speed demodulator, low-pass filter, and multiple-sampling amplifier is designed. The residual offset and low-frequency noise are reduced by two steps; one is sampling phase adjusting in the multiple-sampling amplifier and the other is a double modulation technique. The double modulation effectively reduces the offset and low-frequency noise, or 1/f noise. A SRS CMOS imager chip for the proof of concept is implemented and a successful operation is demonstrated. The detectable AC signal level under the large DC offset is measured to be smaller than ratio of 10^<-4>. This finding suggests that the detection of a very small SRS signal from a biological sample is feasible using this technique.
• 近赤外光・電子顕微鏡による相関バイオイメージング  [Not invited]

  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

  2014/10
• Observation of arteriosclerotic lesion using coherent nonlinear Raman imaging  [Not invited]

  Mamoru Hashimoto, Shoji Kawahito

  Transactions of Japanese Society for Medical and Biological Engineering  2014/08  Japan Soc. of Med. Electronics and Biol. Engineering
   

  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.
• Histological assessment of atherosclerosis by nonlinear optical multimodal microscopy  [Not invited]

  Tomoyo Tao, Harsono Cahyadi, Shuichiro Fukushima, Mamoru Hashimoto, Tsutomu Araki

  Transactions of Japanese Society for Medical and Biological Engineering  2014/08  Japan Soc. of Med. Electronics and Biol. Engineering
   

  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.
• 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  [Not invited]

  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

  2014/07
• Cathodoluminescence and Fluorescent Bioimaging with Using Rare-Earth Doped Nanophorspohrs  [Invited]

  Niioka Hirohiko, Furukawa Taichi, Mamoru Hashimoto

  顕微鏡  2014/04
• カソードルミネッセンス顕微鏡と光学顕微鏡の融合  [Invited]

  新岡宏彦, 古川太一, 福島昌一郎, 一宮正義, 三宅 淳, 芦田昌明, 荒木 勉, 橋本 守

  日本顕微鏡学会バイオメディカルニュ－マイクロスコ－プ分科会 平成25年度シンポジウム講演会  2014/03
• 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  [Not invited]

  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

  2014/03
• Synthesis of nanophosphors for correlative cathodoluminescence, up-conversion, and near-infrared bioimaging  [Not invited]

  S. Fukushima, T. Furukawa, H. Niioka, M. Ichimiya, J. Miyake, M. Ashida, T. Araki, M. Hashimoto

  2014/03
• Visible to near-infrared luminescent nanoparticles for multimodal bioimaging on nanometer to millimeter scale  [Not invited]

  S. Fukushima, T. Furukawa, H. Niioka, M. Ichimiya, J. Miyake, M. Ashida, T. Araki, M. Hashimoto

  Japan Taiwan Bilateral Conference on Biomedical and Plasmonic Imaging  2014/02
• 1F16 Histological assessment of atherosclerosis by nonlinear optical multimodal microscopy

  TAO Tomoyo, FUKUSHIMA Shuichiro, HASHIMOTO Mamoru, ARAKI Tsutomu

  バイオエンジニアリング講演会講演論文集  2014/01  The Japan Society of Mechanical Engineers
  
• Development of nonlinear optical multimodal microscopy system for histological assessment of atherosclerosis

  Tao Tomoyo, Harsono Cahyadi, Fukushima Shuichiro, Hashimoto Mamoru, Araki Tsutomu

  BME  2014  Japanese Society for Medical and Biological Engineering
   

  We developed a nonlinear optical multimodal microscopy system for diagnostic tool of atherosclerosis. Nonlinear optical microscopy is based on the nonlinear optcal phenomena originated from optical properties of materials. It avoids the need for invasive dyes. In particular, coherent Raman scattering (CRS) techniques provide label-free detection due to the intrinsic vibrational signatures of the specific molecules. Therefore it offers a non-invasive imaging of lipid in atherosclerotic plaques. The second harmonic generation (SHG) is exclusively sensitive to noncentrosymmetric materials. Hence, it enables a label-free detection of collagen which is known to be unpredictably formed around the plaques. Therefore, CRS/SHG multimodal microscopy system is a potential tool for quantitative diagnosis of atherosclerosis.
  In this study, we provided a histological assessment of abnormally produced collagen in aorta samples of mice, and our results indicated that the collagen had a correlation with the progression of lesion. Furthermore, we performed CRS/SHG imaging with the developed system.
• B204 Correlative bioimaging with near-infrared and electron microscopy

  Shoichiro FUKUSHIMA, Hirohiko NIIOKA, Masayoshi ICHIMIYA, Masaaki ASHIDA, Jun MIYAKE, Tsutomu ARAKI, Mamoru HASHIMOTO

  The Proceedings of the JSME Conference on Frontiers in Bioengineering  2014  Japan Society of Mechanical Engineers
  
• Lock-in Pixels Readout Circuit Using a High Speed Lateral Electric Field Modulator with Differential Charge Accumulation for Stimulated Raman Scattering Imager  [Not invited]

  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)  2014  IEEE
   

  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.
• 3P-247 Development of near infrared phosphor probes for deep tissue imaging toward the application for regenerative medicine  [Not invited]

  Niioka Hirohiko, Fukushima Syoichiro, Hashimoto Mamoru, Araki Tsutomu, Onoshima Daisuke, Yukawa Hiroshi, Baba Yoshinobu, Miyake Jun

  日本生物工学会大会講演要旨集  2014  日本生物工学会
  
• Tens nanometer scale cathodoluminescence bioimaging with rare-earth doped nanophosphors  [Not invited]

  Shoichiro Fukushima, Hirohiko Niioka, Masayoshi Ichimiya, Masayoshi Ichimiya, Jun Miyake, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto

  JSAP-OSA Joint Symposia, JSAP 2014  2014/01
• Observation of anhydrated and hydrated DAST crystals using multiplex fourth order Raman microscope  [Not invited]

  Chikako Ninagawa, Hirohiko Niioka, Tsutomu Araki, Mamoru Hashimoto

  JSAP-OSA Joint Symposia, JSAP 2014  2014/01
• Multimodal bioimaging probes based on lanthanide doped Gd2O3 nanophosphors  [Not invited]

  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  2014/01
• 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  [Not invited]

  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

  2013/11
• 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  [Not invited]

  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

  2013/11
• Upconversion fluorescence and CL imaging for multiscale biological imaging  [Not invited]

  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  2013/11
• Bimodal biological observation with luminescence emitted under electron beam and near-infrared light irradiation  [Not invited]

  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  2013/11
• A220 Observation of atherosclerosis by second harmonic generation microscopy

  TAO Tomoyo, FUKUSHIMA Shuichiro, HASHIMOTO Mamoru, ARAKI Tsutomu

  Proceedings of the ... JSME Conference on Frontiers in Bioengineering  2013/10  The Japan Society of Mechanical Engineers
  
• 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  [Not invited]

  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

  2013/10
• Upconversion Nanophosphors for Correlative CL and Fluorescent Imaging  [Not invited]

  H. Niioka, T. Furukawa, S. Fukushima, M. Ichimiya, T. Nagata, J. Miyake, M. Ashida, T. Araki, M. Hashimoto

  2013/10
• Towards multicolor correlative light and cathodoluminescence imaging with using upconversion nanophosphors  [Not invited]

  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  2013/09
• 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  [Not invited]

  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

  2013/07
• A High-Speed Modulation Lock-in Pixel Imager Using Differential Charge Accumulation for Stimulated Raman Scattering Spectroscopy

  MARS Kamel, BEAK Guseul, HANG Sang Man, TAKASAWA Taishi, YASUTOMI Keita, KAGAWA Keiichiro, HASHIMOTO Mamoru, KAWAHITO Shoji

  電気学会研究会資料. BMS, バイオ・マイクロシステム研究会 = The papers of Technical Meeting on Bio Micro Systems, IEE Japan  2013/03
• 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  [Not invited]

  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

  2013/03
• 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  [Not invited]

  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

  2013/03
• Synthesis of Rare-earth Doped Nano Phosphors for Biological Cathodoluminescence Imaging  [Not invited]

  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)  2013/03
• C212 Synthesis of bimodal nanophosphors with cathodoluminescence and upconversion luminescence for multi-scale bioimaging  [Not invited]

  Shoichiro FUKUSHIMA, Taichi FURUKAWA, Hirohiko NIIOKA, Mamoru Hashimoto

  The Proceedings of the JSME Conference on Frontiers in Bioengineering  2013  Japan Society of Mechanical Engineers
  
• Rare-earth Doped Y2O3 Nanophosphors Synthesized for Bio-imaging with Using CL and Fluorescence Microscopy  [Not invited]

  H. Niioka, T. Furukawa, M. Ichimiya, T. Nagata, M. Ashida, T. Araki, M. Hashimoto

  8th Handai Nanoscience and Nanotechnology International Symposium  2012/12
• A Draining-Only Modulation CMOS Image Sensor for Fluorescence Lifetime Imaging

  YASUTOMI Keita, LI Zhuo, KAGAWA Keiichiro, NIIOKA Hirohiko, HASHIMOTO Mamoru, KAWAHITO Shoji

  ITE Technical Report  2012/11  The Institute of Image Information and Television Engineers
   

  The paper presents a time-resolved CMOS image sensor with Draining Only Modulation(DOM) pixels for fluorescence lifetime imaging. The proposed DOM pixels, which enables signal charge transfer without any transfer gates, provides high-speed charge modulation and repetitive accumulation at very low light level. The prototype imager demonstrates loss-free charge accumulation and fluorescence lifetime measurements for different fluorescent samples.
• 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  [Not invited]

  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

  2012/11
• OS3-1-3 Cellular imaging with using cathodoluminescence and rare earth doped nanophosphors

  Furukawa Taichi, Niioka Hirohiko, Ichimiya Masayoshi, Nagata Tomohiro, Ashida Masaaki, Araki Tsutomu, Hashimoto Mamoru

  マイクロ・ナノ工学シンポジウム  2012/10  The Japan Society of Mechanical Engineers
   

  Bio molecular imaging is important to clarify cellular functions. Cathodoluminescence (CL) is light emission from the materials excited by accelerated electron beam, and CL microscopy has the potential to enable color imaging of individual biomolecular distributions with using immunolabeling at high spatial resolution. Because of electron beam excitation the spatial resolution reaches about 10 nm. In this study, we demonstrated CL imaging for cells using rare-earth doped nanophosphors at high spatial resolution.
• 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  [Not invited]

  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

  2012/09
• Multimodal Imaging via Light Microscopy and Cathodoluminescence Microscopy for Biological Specimens with Rare-earth Doped Nanophosphors  [Not invited]

  T. Furukawa, H. Niioka, M. Ichimiya, T. Nagata, M. Ashida, T. Araki, M. Hashimoto

  JSAP-OSA joint Symposia 2012, the 73th Autumn Meeting  2012/09
• 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  [Invited]

  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)  2012/05
• Well-dispersed nanophosphors for cathodoluminescence microscopy produced by using laser ablation method  [Not invited]

  T. Furukawa, H. Niioka, M. Ichimiya, T. Nagata, M. Ashida, T. Araki, M. Hashimoto

  Program and Abstract Book, Focus on Micrscopy 2012 (FOM2012)  2012/04
• 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  [Not invited]

  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

  2012/03
• 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  [Not invited]

  新岡宏彦, 古川太一, 一宮正義, 芦田昌明, 荒木 勉, 橋本 守

  2012/02
• 7D32 Intracellular imaging of anticancer drug using CARS microscopy

  IKEDA Kouhei, CAHYADI Harsono, NIIOKA Hirohiko, ARAKI Tsutomu, HASHIMOTO Mamoru

  バイオエンジニアリング講演会講演論文集  2012/01  The Japan Society of Mechanical Engineers
  
• 7D45 Observation of biological molecules using hyper-Raman microspectroscopy

  KANOH Hiroto, NIIOKA Hirohiko, ARAKI Tsutomu, HASHIMOTO Mamoru

  バイオエンジニアリング講演会講演論文集  2012/01  The Japan Society of Mechanical Engineers
  
• 7D44 Distinction between triglyceride deposit cardiomyovasculopathy and ischemic cardiomyopathy with Raman microscopy and multivariable analysis

  MATSUMURA Naokazu, NIIOKA Hirohiko, IKEDA Yoshihiko, HIRANO Ken-ichi, ARAKI Tsutomu, HASHIMOTO Mamoru

  バイオエンジニアリング講演会講演論文集  2012/01  The Japan Society of Mechanical Engineers
  
• 7D31 Development of CARS microscopy using picosecond high speed wavelength scanning laser  [Not invited]

  IWATSUKA Junichi, MINAMIKAWA Takeo, NIIOKA Hirohiko, ARAKI Tsutomu, HASHIMOTO Mamoru

  The Proceedings of the Bioengineering Conference Annual Meeting of BED/JSME  2012  The Japan Society of Mechanical Engineers
  
• 7D46 Photodamage to cells in multi-focus nonlinear Raman microscopy  [Not invited]

  MURAKAMI Yoshinori, MINAMIKAWA Takeo, ARAKI Tsutomu, HASHIMOTO Mamoru

  The Proceedings of the Bioengineering Conference Annual Meeting of BED/JSME  2012  The Japan Society of Mechanical Engineers
  
• Super-resolution imaging of nano phosphors via cathodoluminescence microscopy for biological imaging  [Not invited]

  H. Niioka, T. Furukawa, M. Ichimiya, M. Ashida, T. Araki, M. Hashimoto

  2011/12
• 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  [Not invited]

  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

  2011/11
• Super-Resolution Imaging for Cells by using Cathodoluminescence  [Not invited]

  T. Furukawa, H. Niioka, M. Ichimiya, M. Ashida, T. Araki, M. Hashimoto

  2011/09
• 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  [Not invited]

  古川太一, 新岡宏彦, 一宮正義, 芦田昌明, 荒木 勉, 橋本 守

  2011/07
• T0302-2-3 Noncontact and nondestructive measurement of strain using three-dimensional molecular orientation measurement under microscope

  YOSHIKI Keisuke, Masuda Kyosuke, HASHIMOTO Mamoru, HASHIMOTO Nobuyuki, KURIHARA Makoto, Namazu Takahiro, INOUE Shozo

  The proceedings of the JSME annual meeting  2010  The Japan Society of Mechanical Engineers
   

  We developed a system to analyze microscopic deformation, which can visualize inhomogeneous deformation in MEMS(Micro Electro Mechanical Systems) devices by using single-molecule markers. As MEMS devices become smaller, their mechanical properties seem to become inhomogeneous. This causes an undesirable distribution of deformation and stress, and decreases the reliability of the MEMS devices. We distributed fluorescent single molecules discretely on the microstructure of a MEMS device, and traced their relative displacements and rotations by observation under a three-dimensional orientation microscope, which provided information about normal, torsional, and bending displacements.
• B201 Label-free and real-time CARS imaging of living cell reactions in laser-induced ablation  [Not invited]

  MINAMIKAWA Takeo, NIIOKA Hirohiko, ARAKI Tsutomu, HASHIMOTO Mamoru

  The Proceedings of the JSME Conference on Frontiers in Bioengineering  2009  The Japan Society of Mechanical Engineers
  
• Realtime biomolecular imaging by multifocus CARS microscopy  [Not invited]

  2009
• High-speed non-staining biomolecular imaging by nonlinear Raman microspectroscopy  [Not invited]

  2009
• Non-staining Visualization of Collagen by Second Harmonic Generation Microscopy

  FUKUSHIMA Shuichiro, YASUI Takeshi, HASHIMOTO Mamoru, ARAKI Tsutomu

  可視化情報学会誌. Suppl.  2008/07
• 1203 Second harmonic generation imaging of organic molecule on metal by radial polarization excitation

  Ashida Koichiro, Yoshiki Keisuke, Hashimoto Mamoru, Araki Tsutomu

  関西支部講演会講演論文集  2008/03  The Japan Society of Mechanical Engineers
  
• 602 Label-free, high-speed imaging with real-time CARS microscope  [Not invited]

  MINAMIKAWA Takeo, HASHIMOTO Mamoru, ARAKI Tsutomu

  The Proceedings of Conference of Kansai Branch  2008  The Japan Society of Mechanical Engineers
  
• 329 Development of second-harmonic-generation microscope to determine 3-dimensional molecular orientation with 3-dimensional spatial resolution

  YOSHIKI Keisuke, HASHIMOTO Mamoru, ARAKI Tsutomu

  バイオエンジニアリング講演会講演論文集  2006/01  The Japan Society of Mechanical Engineers
  
• Three dimensional polarization control and its application to SHG imaging  [Not invited]

  K. Yoshiki, M. Hashimoto, T. Araki

  2005 Pacific Rim Conference on Lasers & Electro-Optics  2005/07  IEEE
  
• Multi-focus CARS microscopy using automatic pulse duration control system  [Not invited]

  Proceedings of International Conference on Quantum Electronics 2005 and the Pacific Rim Conference on Lasers and Electro-Optics 2005 (IQEC/CLEO-PR 2005)  2005
• 金属プローブを用いた近接場CARSイメージング  [Not invited]

  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  2004/05
• 局所プラズモン増強効果を用いた高空間分解CARSイメージング  [Not invited]

  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  2004/03
• チップ増強非線形ラマン散乱を用いた近接場振動分光  [Not invited]

  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  2004/03
• Coherent anti-stokes Raman nano-imaging with metal-tip field enhancement  [Not invited]

  N Hayazawa, T Ichimura, M Hashimoto, Y Inouye, S Kawata

  ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY  2004/03  AMER CHEMICAL SOC
  
• Coherent anti-Stokes Raman microscope for identification of cellular molecule  [Not invited]

  Proceedings of the Second Asian and Pacific Rim Symposium on Biophotonics 234-235  2004
• Fluorescence lifetime properties of various calcium ion indicators  [Not invited]

  Proc. ICO-14  2004
• 金ナノ微粒子によるコヒーレントアンチストークスラマン散乱の局所増強  [Not invited]

  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  2003/07
• 金属ナノ微粒子による局所増強コヒーレントアンチストークスラマン散乱  [Not invited]

  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  2003/03
• Coherent anti-Stokes Raman scattering microscopy using near IR exitation and UV excitation

  Ueda Kei, Hashimoto Mamoru, Araki Tsutomu, Kawata Satoshi

  バイオエンジニアリング講演会講演論文集  2003/01  The Japan Society of Mechanical Engineers
  
• Confocal Fluorescence Lifetime Imaging by Asynchronous Sampling

  AZUMA Hiroki, INAMOTO Hirohisa, HASHIMOTO Mamoru, ARAKI Tsutomu, CHIKAMORI Kensuke

  関西支部講演会講演論文集  2003  The Japan Society of Mechanical Engineers
   

  We have developed a confocal microscope for fluorescence lifetime imaging by asynchronous sampling. This system consists of a fast response photomultiplicr, a fast sampling module, and a low-pass filter and a colnfocal microscope. Since imaging by laser scanning and detection is asynchronous with laser excitation. It is possible to attach the detection system to the commercial confocal microscopy system. We also observed the relationship between the concentration of Ca^<2+> and the fluorescence lifetime of fluo-3,and confirmed that the fluorescence lifetime is sensitive to the concentration of Ca^<2+>.
• Coherent anti-Stokes Raman scattering microscopy

  HASHIMOTO Mamoru

  Electron-microscopy  2002/11  日本電子顕微鏡学会
  
• Coherent Anti-Stokes Raman Scattering Microscopy

  HASHIMOTO Mamoru, ARAKI Tsutomu, KAWATA Satoshi

  IEICE technical report. ME and bio cybernetics  2002/10  The Institute of Electronics, Information and Communication Engineers
   

  New multiphoton microscopy using CARS (coherent anti-Stokes Raman scattering) spectroscopy is described. In CARS microscopy, the image of molecular vibration that is sensitive to the molecular spices and conformation is obtainable without staining with three-dimensional resolution of sub-micrometer. 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.
• Resonance enhancement of coherent anti-Stokes Raman scattering microscopy

  Ueda Kei, Hashimoto Mamoru, Araki Tsutomu, Kawata Satoshi

  Proceedings of the JSME Bioengineering Conference and Seminar  2002  The Japan Society of Mechanical Engineers
  
• Capillary electrophoresis system using a fluorescence labeled cell as a sensor probe

  FURUMOTO Masatsugu, SATO Tatsuya, HASHIMOTO Mamoru, YASUI Takeshi, ARAKI Tsutomu

  バイオエンジニアリング講演会講演論文集  2001/01  The Japan Society of Mechanical Engineers
  
• Molecule Vibrational Imaging by CARS microscopy

  HASHIMOTO Mamoru, INOUE Keigo, FUJITA Katsumasa, NAKAMURA Osamu, KAWATA Satoshi, ARAKI Tsutomu

  バイオエンジニアリング講演会講演論文集  2001/01  The Japan Society of Mechanical Engineers
  
• Multi-spectral imaging by multi-focus CARS microscopy  [Not invited]

  Proceedings of Multi-dimensional Microscopy 2001, 23  2001
• 1117 Change of Ca content in tissue of human blood vessel with aging and region

  ARAKI Tsutomu, TATEYAMA Yutaka, HASHIMOTO Mamoru, MASUDA Mitsuhiko, TOHNO Yoshiyuki

  関西支部講演会講演論文集  2000/03  The Japan Society of Mechanical Engineers
   

  We have determined Ca contents in human vessels by Ar-microwave induced plasma atomic emission spectroscopy. 0n the basis of this measurement, we have studied relation between Ca content and age, and that of between Ca content and vessel region. Ca contents are femoral artery > thoracic aorta > internal jugular vein > superior vena cava > inferior vena cava. In the case of arterial tissue, Ca content increases with aging. Ca contents in umbilical artery and umbilical vein, which are used as the reference to aging and mechanical stress, are the same level as those in adult veins. Ca contents in monkey arteries are much smaller than those in human vessels, and the difference between Ca content in upper limb and that in lower one is not evident. In the case of human, however, Ca content in artery of lower limb is larger than that of upper one, and especially large in the part of elbow, thigh and knee. These results suggest that the effect of mechanical stress is one important factor for increase of Ca content in the arterial tissue.
• Capillary Electrophoresis System using Biological Reaction of Single Cell as a Sensor Probe

  SATO Tatsuya, HASHIMOTO Mamoru, YASUI Takeshi, ARAKI Tsutomu

  The proceedings of the JSME annual meeting  2000  The Japan Society of Mechanical Engineers
   

  We have constructed a capillary electrophoresis (CES) system using living cells as the sensor tip of CES to monitor the change of cellar environment. In the present system, yeast cell stained with fluorescence dye was applied as the sensor cell. Fluorescence intensity of the probe cell changes according to the change of cellar environment. To confirm the performance of the proposed CES system, the pollutant (Hg^<2+>) and the biological substance (acetylcholine) were applied. Fluorescence intensity of Rhodamine123 (membrane potential sensitive dye for mitochondria), decreased gradually due to effect of the pollutant, while that of Fluo-3 (calcium sensitive dye) increased.
• Changes of autofluorescence in human dentine caused by caries

  NAKAMOTO Kazunobu, YASUI Takeshi, HASHIMOTO Mamoru, ARAKI Tsutomu

  Proceedings of the JSME Bioengineering Conference and Seminar  2000  The Japan Society of Mechanical Engineers
  
• Effect of the detector side pinhole on 3D optical properties of CARS microscopy  [Not invited]

  Proceedings of Focus on Microscopy 2000  2000
• Molecular Vibrational Imaging in the Fingerprint Region by Cars Microscopy  [Not invited]

  Proceedings of Focus on Microscopy 2000, p.~4  2000
• 同期式ナノ秒Xeアークランプ低ジッター化とその応用  [Not invited]

  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  1997

MISC

• Tens nanometer scale cathodoluminescence bioimaging with rare-earth doped nanophosphors

  Shoichiro Fukushima, Hirohiko Niioka, Masayoshi Ichimiya, Masayoshi Ichimiya, Jun Miyake, Masaaki Ashida, Tsutomu Araki, Mamoru Hashimoto  JSAP-OSA Joint Symposia, JSAP 2014  2014/01  [Not refereed][Not invited]
  
• Observation of anhydrated and hydrated DAST crystals using multiplex fourth order Raman microscope

  Chikako Ninagawa, Hirohiko Niioka, Tsutomu Araki, Mamoru Hashimoto  JSAP-OSA Joint Symposia, JSAP 2014  2014/01  [Not refereed][Not invited]
  
• 3P-247 Development of near infrared phosphor probes for deep tissue imaging toward the application for regenerative medicine

  Niioka Hirohiko, Fukushima Syoichiro, Hashimoto Mamoru, Araki Tsutomu, Onoshima Daisuke, Yukawa Hiroshi, Baba Yoshinobu, Miyake Jun  日本生物工学会大会講演要旨集  66-  256  -256  2014  [Not refereed][Not invited]
  
• Multimodal bioimaging probes based on lanthanide doped Gd2O3 nanophosphors

  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  2014/01  [Not refereed][Not invited]
  
• 生体相関イメージングを目指したCUPLナノ蛍光体の合成

  福島 昌一郎, 橋本 守  ホソカワ粉体工学振興財団年報  22-  117  -120  2014
• Towards multicolor correlative light and cathodoluminescence imaging with using upconversion nanophosphors

  S. Fukushima, T. Furukawa, H. Niioka, M. Ichimiya, M. Ichimiya, T. Nagata, J. Miyake, M. Ashida, T. Araki, M. Hashimoto  Optics InfoBase Conference Papers  2013/12  [Not refereed][Not invited]
  
• 7D31 Development of CARS microscopy using picosecond high speed wavelength scanning laser

  IWATSUKA Junichi, MINAMIKAWA Takeo, NIIOKA Hirohiko, ARAKI Tsutomu, HASHIMOTO Mamoru  The Proceedings of the Bioengineering Conference Annual Meeting of BED/JSME  2012-  (0)  _7D31  -1_-_7D31-2_  2012  [Not refereed][Not invited]
  
• 7D46 Photodamage to cells in multi-focus nonlinear Raman microscopy

  MURAKAMI Yoshinori, MINAMIKAWA Takeo, ARAKI Tsutomu, HASHIMOTO Mamoru  The Proceedings of the Bioengineering Conference Annual Meeting of BED/JSME  2012-  (0)  _7D46  -1_-_7D46-2_  2012  [Not refereed][Not invited]
  
• B201 Label-free and real-time CARS imaging of living cell reactions in laser-induced ablation

  MINAMIKAWA Takeo, NIIOKA Hirohiko, ARAKI Tsutomu, HASHIMOTO Mamoru  The Proceedings of the JSME Conference on Frontiers in Bioengineering  2009-  (0)  93  -94  2009  [Not refereed][Not invited]
  
• 602 Label-free, high-speed imaging with real-time CARS microscope

  MINAMIKAWA Takeo, HASHIMOTO Mamoru, ARAKI Tsutomu  The Proceedings of Conference of Kansai Branch  2008-  (0)  _6  -2_  2008  [Not refereed][Not invited]
  
• 研究費を獲得しよう

  橋本 守  光学  31-  (11)  843  -844  2002/11/10
• Optical Measurements

  HASHIMOTO Mamoru  光学  31-  (5)  395  -395  2002/05/10

Industrial Property Rights

• 特許6817623:Polarization control device and polarization control method    2021/01/20

  Keisuke Yoshiki, Mamoru Hashimoto  University of Hyogo
• 特許6425242:変調光検出のＳＮ比を向上する方法    2018/11/21

  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    2016/11/30

  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    2016/09/06

  新岡 宏彦, 橋本 守, 古川 太一  国立大学法人大阪大学
• 特許5926055:波長走査パルス光同期システムおよびその制御方法  2016/05/25

  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    2015/12/09

  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    2012/01/25

  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

Awards & Honors

• 2014/07 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
• 2010/02 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
• 2009/08 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
• 2009/03 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
• 2006/01 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
• 2002/05 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

Research Grants & Projects

• Advanced non-invasive optical measurement techniques for the constant monitoring of blood lipid components

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2023/04 -2026/03 

  Author : 加藤 祐次, 清水 孝一, 松村 健太, 橋本 守
• Near-infrared light delivery in deep tissue using vivo helical optical waveguide formed by a topological ultrasound beam

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2022/06 -2025/03 

  Author : 橋本 守, 工藤 信樹, 加藤 祐次
• Development of coherent Raman scattering microendoscope and application to unstained in-situ diagnosis

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)

  Date (from‐to) : 2021/04 -2024/03 

  Author : 橋本 守, 七戸 俊明, 新岡 宏彦

   

  色素の静脈投与なしに癌を内視鏡検査で確定診断するために，上部消化管内視鏡(胃カメラ)の 鉗子孔に挿入可能な非線形ラマン散乱プローブ顕微内視鏡の開発を目指している．非線形ラマン散乱の一種であるコヒーレント反ストークスラマン散乱（CARS）を用いることで，無染色・高空間分解能・高速な細胞，細胞核の形状観測を可能とする．本年は，CARSプローブ顕微内視鏡におけるキーデバイスである，2波長板（DWP）の設計を行なった．DWPは，直交する偏光を持つ２波長の励起光を，互いに平行な偏光へと変換するデバイスである．このために，709 nmnの励起光に対してはリタデーションをゼロとし，888 nmの励起光に対してはλ/2となるように設計する必要がある．２波長に対してこのような特性を持つ波長板は，2種類の１軸性結晶を用いれば可能であるが高コストとなるため，1種類の結晶で設計可能かシミュレーションした．結晶には水晶を用い，発生するCARS光強度を最大となるように結晶の厚さを最適化したところ，0.1980 mmで，完全に平行な光を用いて励起した際の99%のCARS信号が得られることが分かった．また，同様に90%以上のCARS信号が得られるラマンシフト領域は，2,530 to 2,980 cm-1であることが分かった．したがって，このDWPでCH伸縮振動に対応する高波数領域のCARS信号を十分取得できることが分かった．また，励起光源としてファイバーレーザーベースの光源を作成し1550 nmでのパルス発振を確認できた．これまでに開発してきた原理検証システムを用いて，ポリスチレンビーズや培養細胞やラット組織の観測を行なった．ポリスチレンビーズの観測は行えたが，培養細胞や組織のCARS信号を得ることはできなかった．
• Development of transcutaneous optical measurement of blood particulate concentration for non-invasive monitoring of blood components

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2020/04 -2023/03 

  Author : 加藤 祐次, 清水 孝一, 橋本 守, 松村 健太

   

  本研究の大きな目標である血中中性脂肪濃度の無侵襲光学計測の実現のため、計測手法は生体透過性の高い可視から近赤外の波長領域での後方散乱光の空間分解計測を基礎としている。前年度まで、計測対象の一つとしている前腕部静脈の数値モデルに対するモンテカルロ・シミュレーションにより、静脈の形状・位置に対して光源と検出器の距離を調整する必要性が分かったため、センサー配置の最適化について検討した。また、推定アルゴリズム精度のさらなる向上のため、静脈が走行していない箇所での周囲媒質のみの光学特性推定を組み合わせて、個人差や計測箇所による変動を抑えたロバスト性の向上を進めた。さらに、メディカルフォトニクス社の協力により試作機を用いて、これまで準備してきた実験ファントムを用いて実測し、シミュレーションや光拡散理論と比較検討を行った。その結果シミュレーションで検討してきた推定アルゴリズムに対して修正指針を得た。 一方、毛細血管を対象とした光電式脈波計測法の応用について、スマートフォンアプリおよび多色のLEDとPDからなる計測モジュールを用い、前年度までの検討を踏まえて接触圧力をコントロールしつつ、脂肪食摂取前後の光電脈派信号の変化について評価した。その結果、現状では脂質濃度変化に対する光電脈波信号の変化は見出すことはできなかった。今後、脈波信号の解析について単純な信号強度の変化のみではなく、周波数解析等による解析を要することが課題となった。さらに、ハードウェア面で通常の光電脈波計測のみならず、干渉法に基づくスペックル計測法の適用等の計測法の拡張についても今後の課題とし、基礎実験データを得た。
• 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

  Date (from‐to) : 2020/06 -2021/05
• Artificial intelligence pathological diagnosis by hyperspectral nonlinear Raman scattering imaging

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2017/04 -2020/03 

  Author : Hashimoto Mamoru

   

  By reducing the noise of observation data and improvement of observation time using deep learning, we succeeded in the imaging rate improvement of 1.6–1.2 image/min. to 12.5-4.0 image/min. In tissue classification by machine learning of nonlinear Raman scattering images, pre-training with fluorescence images significantly improved the segmentation ability. Besides, we have developed a new microscope for acquiring a large number of hyperspectral nonlinear Raman scattering images and succeeded in increasing the speed 14 times and reducing the excitation light peak irradiance 1/12 compared to the conventional method. It was shown that hyperspectral images of cultured cells were classified by deep learning, and that unsupervised learning could classify cells with different culture conditions.
• Non-invasion observation of the cell culture process by nonlinear optical microscopy

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2015/04 -2019/03 

  Author : FUKUSHIMA Shuichiro

   

  The importance of controlling microscopic culture environment in cellular scale, was verified using microfluidic devices. By using an oxygen concentration controlled device, it was possible to realize the co-culture of the cells in different oxygen environment, which is difficult to control by the conventional methods. Moreover, the usefulness of nonlinear optical microscopy was shown as a method of non-invasion observation of the cell culture process. Visualization and deformation analysis of the substrate structure using second harmonic generation light are effective for the examination of the mechanical environmental factor, when the collagen gel is made to be culture substrate.
• Super-resolution Imaging of nonlinear Raman atomic force imaging

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Challenging Research (Exploratory)

  Date (from‐to) : 2017/06 -2019/03 

  Author : Hashimoto Mamoru

   

  We have developed a super-resolution molecular vibration imaging microscope that combines atomic force microscopy (AFM) with nonlinear Raman scattering beyond the diffraction limit of light. The volume expansion accompanying the molecular vibrational excitation via nonlinear Raman scattering process is observed by AFM. In order to focus the laser beam near the probe of AFM, it is necessary to detect volume expansion with high sensitivity while irradiation of the probe. Therefore, we developed a system in which the frequency difference between the pulse oscillations of the two lasers is synchronized with the resonance of the cantilever so that the two pulse lights are simultaneously irradiated to the sample only when the probe contacts the sample. Using the developed device, we successfully detected a signal that seems to be volume expansion induced by nonlinear Raman scattering.
• Elucidation of the glycated collagen reaction in dentin and its application to next dental treatment

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2016/04 -2019/03 

  Author : Miura Jiro, Araki Tsutomu

   

  Granular calcification is formed in the dental pulp of diabetic patients. Histopathologically, these are thought to be by-products of severe diabetes. In this study, the pathological calcification in dental pulp of Type2 diabetes model rat was evaluated using electron microscopy and immunohistochemical analysis of pentosidine, a kind of Advanced Glycation End products (AGEs) which is thought to be related calcification .The composition of pulp stone was examined with Energy Dispersive X-ray Spectoroscopy. In the type2 diabetes model rats, it was found that pathogenic calcification in pulp occurs with the high blood glucose level for 10 weeks in SDT fatty rats. Pentosidine which is a kind of AGEs strongly expresses in the pulp of diabetic rats. And granular calcification was also observed in the surrounding area of pulp stone. From these results, Glycation was related with calcification in dental pulp, suggesting that these are related to ectopic calcification in diabetic patients.
• Development of fluorescence lifetime measurement for glycation screening for diabetes in vivo

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2016/04 -2018/03 

  Author : Miura Jiro

   

  Glycation, namely the Maillard reaction, is a non-enzymatic reaction between protein or lipid and reducing sugar, resulting in formation of advanced glycation end-products (AGEs). Because glycation of collagen progresses under physiological conditions in living organisms, glycation is one of the most important processes in human aging. Several AGEs bind collagen and act as cross-links between collagen fibrils, thereby changing the fluorescent lifetime of collagen-rich tissues. The purpose of this study is to identify AGEs in human dentin using electron microscopy, time-resolved fluorescence microscopy, mechanical testing and chemical analyses. Our findings suggest the potential influence of cross-linking on diabetic and aging tissue.
• Frequency-swept discrete Fourier transform spectroscopy ~demonstration and application in THz and near-infrared region~

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)

  Date (from‐to) : 2014/04 -2017/03 

  Author : YASUI TAKESHI

   

  In this research, we demonstrated improved spectral resolution by overcoming the time window size limitation using a mode-locked terahertz (THz) or near-infrared (NIR) 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 only limited by the spectral interleaving interval.
• 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」

  Date (from‐to) : 2010/12 -2017/03 

  Author : 橋本 守
• Crystallization monitoring using higher order nonlinear Raman scattering microscopy

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2013/04 -2016/03 

  Author : Hashimoto Mamoru, FUKUSHIMA Shuichiro, NIIOKA Hirohiko

   

  We propose to use fourth order coherent Raman scattering (FOCRS) microspectroscopy for monitoring the crystallization process. The crystallization is a process in the saturated solution. The even order nonlinear phenomenon is prohibited from the solution and provides only the information of crystallized part. FOCRS also provides the information of crystal and the molecular information by the molecular vibration. We have developed a multiplex FOCRS microspectroscopy system using supercontinuum light generated by a photonic crystal fiber. We succeed in observation of FOCRS hyper-spectral image of a DAST (4-dimethylamino-N-methyl-4-stilbazolium tosylate) crystal in saturated solution. We also observed FOCRS and CARS (coherent anti-Stokes Raman scattering) hyper-spectral images of hydrated and anhydrated DAST crystals. CARS bands are clearly observed for both of crystals, but only the anhydrated DAST crystals show FOCRS bands. As a result, the selectivity of FOCRS was declared.
• Multimodal molecular imaging system and its application for investigation of in vivo cell/tissue

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)

  Date (from‐to) : 2011/04 -2014/03 

  Author : ARAKI Tsutomu, HASHIMOTO Mamoru, FUKUSHIMA Shuichiro, NIIOKA Hirohiko

   

  Aims of this research project are development and application of novel imaging method that can informs molecular morphology and function of biological specimens. In the present research, we have focused on the second harmonic generation (SHG) microscopy and cathode luminescence(CL) microscopy.A polarization resolved, high-speed SHG microscope was developed. In in vivo measurement with this microscope, age related dermis collagen image were observed. Also we demonstrated SHG imaging of tissue-engineered cartilage composed of rabbit chondrocytes and type I collagen gel. From this, we confirmed that it is suitable for evaluating the quality of tissue-engineered cartilage. In the CL microscopy, three kinds of rare-earth-doped nanophosphors were synthesized and injected into macrophages. The spatial distribution of nanophosphors was visualized by using a scanning electron microscope CL system.
• 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：

  Date (from‐to) : 2011/04 -2013/03 

  Author : 橋本 守
• Realtime CARS imaging of cellular response by stimulation of laser ablation

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2010 -2012 

  Author : HASHIMOTO Mamoru, FUKUSHIMA Shuichiro, NIIOKA Hirohiko

   

  We have developed a multifocus coherent anti-Stokes Raman scattering microscopy system combined with a laser-beam ablation system. The developed system visualizes sample response by laser stimulation without staining at vide-rate. The laser beam for ablation is controlled independently of observation. The developed system was applied to cellular response by membrane disruption and crystallizationmonitoring by photon pressure.
• Development and application of real-time molecular microscope based on non-linear optical effect for biological specimen

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)

  Date (from‐to) : 2008 -2010 

  Author : ARAKI Tsutomu, HASHIMOTO Mamoru, FUKUSHIMA Shuichirou, KINOーOKA Masahiro, IWATA Tetsuo, YASUI Takeshi

   

  Aims of this research project are to develop new microscopes utilizing non-linear optical effect induced by an ultra-short pulse laser and to observe molecular imaging of the biological specimen without unwanted staining treatment. We have developed a high-quality SHG microscope and CARS microscope, and following results have been obtained using these microscopes : (1) Age-dependency of collagen distribution in human skin was observed with the SHG microscope. This microscope was also applied to evaluate quality of cartilage cultured in collagen gel. (2) CARS image of Hela cell was demonstrated. Also repair of cell membrane was traced in time based on the CARS images.
• Nonlinear Raman microscopy to detect membrane protein

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2007 -2009 

  Author : HASHIMOTO Mamoru, FUKUSHIMA Shuichiro, FUJITA Katsumasa, YOSHIKI Keisuke

   

  We developed a multi-focus excitation nonlinear Raman microscope using a microlens array scanner for real-time molecular imaging. Parallel exposure of a specimen with light from two highly controlled ps mode-locked lasers (jitter of 30fs, point-by-point wavelength scan within 300ms) and parallel detection with an image sensor enabled real-time imaging. We demonstrated real-time Raman imaging of living cells (frame rate of 10fps). We also combined the developed system with the laser ablation to destruct the cell membrane, and demonstrated to observe the membrane repairing process. We added the polarization pattern control system to a nonlinear Raman microscopy system to detect membrane proteins by control the direction of electro-magnetic filed on the focus spot. The possibility of the selective detection of ordered molecules was confirmed with the imaging of liquid crystal.
• 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

  Date (from‐to) : 2006/01 -2008/12 

  Author : 橋本 守
• Nano-spectroscopy of molecular -metal complex

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2007 -2008 

  Author : INOUYE Yasushi, VERMA Prabhat, HASHIMOTO Mamoru, FUJITA Katsumasa

   

  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

  Date (from‐to) : 2006/04 -2007/03 

  Author : 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

  Date (from‐to) : 2006 -2007 

  Author : 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

  Date (from‐to) : 2006 -2007 

  Author : 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.
• Development of staining-free molecular imaging microscope for biological molecules and its application for quality control of cultured cells

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)

  Date (from‐to) : 2005 -2007 

  Author : ARAKI Tsutomu, HASHIMOTO Mamoru, KINO-OKA Masahiro, YASUI Takeshi, IWATA Tetsuo, TOHNO Yoshiyuki

   

  Aims of this research project are to develop new microscopes by which molecular imaging of the biological specimen can be obtained without unwanted staining treatment, and to apply these microscopes for quality control of the cultured cells. The results are written as follows. (1) We have developed a high-speed SHG microscope and measured in situ tomographic images of collagen molecules in human dermis through the skin. The measurement time to obtain one image was shortened to one second that value is about 1/60 of the conventional equipment. (2) We have developed another SHG microscope with a controlled polarization pattern to detect 3D / molecular orientation. Using this microscope, we have demonstrated the 3D collagen profiles in Achilles tendon. (3) Deformation and remodeling of collagen gel due to endotherial cells have been observed by SHG microscopy in order to apply that microscopy for quality control of the cell culture. (4) We have developed a high-speed CARS microscopy system. Using this system, a real-time imaging of liposomes has been demonstrated. (5) We have measured calcium contents in various vascular walls and analyzed the heterogeneity of the contents based on biomechanical effect. (6) A new quantification technique for calcium ion concentration has been proposed based on fluorescence lifetime measurement. The waveform distortion caused by the frequency band width limitation of the instruments has been compensated by a newly proposed algorithm.
• 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

  Date (from‐to) : 2005/09 -2006/08 

  Author : 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

  Date (from‐to) : 2005/06 -2006/08 

  Author : 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

  Date (from‐to) : 2006/01 -2006/03 

  Author : 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

  Date (from‐to) : 2003/04 -2005/03 

  Author : 橋本 守
• Van der Waals'force-controlled nano Raman scattering spectroscopy

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2004 -2005 

  Author : INOUYE Yasushi, HASHIMOTO Mamoru, FUJITA Katsumasa

   

  We found spectral shift of specific Raman band of carbon nanotubes (CNTs) when van der Waals's force is applied onto the CNTs. We measured Raman spectrum of single CNT bundle while applying stress on it with silver coated cantilever tip of atomic force microscope, and observed spectral change of Raman band of CNT in situ. When we apply stress from 0nN to 2.4nN successively, lower peak of G-band (〜1600cm-1) was red-shifted by 18cm-1. G-band reflects electric property of CNT and has two adjacent peaks. On the other hand, no peak shift was observed in higher peak of G-band. This phenomenon originates from the fact that two peaks are corresponding to the vibrational modes of different vibrational direction, and these two vibrational modes have different shearing stresses with respect to each other. Furthermore, we observed peak shift of radial breathing mode (RBM) around 200cm-1〜300cm-1 at most 5cm-1. RBM depends on the diameter of CNT. This peak shift can be attributed to the fact that the cross-section of CNT is deformed into ellipse shape by applied uniaxial stress on the CNT. We also observed intensity of all Raman band are enhanced when applying stress. This suggests that the band gap energy of CNT comes near to the resonance energy(2.33eV) because of the deformation of CNT along the long axis of CNT, thus contribution of resonant Raman effect is increased. We also performed vibrational calculation of molecule-metal cluster including a number of metal atom based on density functional theory. The calculation showed the experimentally observed band shift by complex formation mechanism is well reproduced if the metal cluster contains at least four Ag atoms.
• 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

  Date (from‐to) : 2004 -2005 

  Author : 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.
• Development of molecular imaging microscope for biological specimen and its application for tissue examination

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2003 -2004 

  Author : ARAKI Tsutomu, HASHIMOTO Mamoru, YASUI Takeshi, TOHNO Yoshiyuki, IWATA Tetsuo

   

  Aim of this research project is to develop an effective method that can sense objective biomolecles without staining and to apply this method to identification of the situation of the molecles. For ths purpose, we have focused on non-linear optical effect in the tissue that is induced by the irradiation with high-intensity very short duration pulsed laser light such as a femtosecond laser. In the actual research, we have developed three microscopes ; fluorescence lifetime imaging microscope in nanosecond range, a collagen sensitive microscope using second harmonic generation light (SCG light) and a coherent anti Stokes coherent Roman scattering (CARS) microscope. The most successful one is the SHG microscope. Based on the reflection-type microscope with a probe light spot of 10 micrometers in diameter and the polarization measurement of SHG light induced by collagen molecules, we have determined distribution of collagen fiber orientation in various human tissues. The resulting SHG data in human skin imply that the reticular dermis posses a tangled structure of collagen fibers which is consistent with the result of the anatomical examination of the skin. We have improved the SHG microscope to obtain 3D dissection image using the con-focal optics and the 3D sample scanning stage. With this microscope, tomograph of the collage fiber orientation in the human skin and bone have been measured. Performance of the fluorescence lifetime imaging microscope was demonstrated by the measurement of the autofluorescence in liposome. Also we have measured the distribution of hyaluronic acid around an erythrocyte with CARS microscope.
• 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)

  Date (from‐to) : 2003 -2004 

  Author : 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

  Date (from‐to) : 2001/04 -2003/03 

  Author : 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

  Date (from‐to) : 2001/04 -2003/03 

  Author : 橋本 守
• Development and application of a novel capillary chromatography system utilizing vital reaction of a single cell

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2001 -2002 

  Author : ARAKI Tsutomu, TOHNO Yoshiyuki, YASUI Takeshi, HASHIMOTO Mamoru, CHIKAMORI Kensuke

   

  We have proposed a biosensor chromatography system in conjunction with a capillary electrophoresis (CE) chromatography and a living cell to monitor the cellular environment. As the sensor cell for bio-CE system, Saccharomyces cerevisiae (SC) was used, SC was stained with rhodamine 123 (Rhl23) that is sensitive to electric potential of the mitochondria membrane. The cell was enveloped inside the capillary, and resultant Rh123 fluorescence image was measured with a fluorescence microscope and a CCD system. In order to avoid unwanted electric field of 5 kV to the sensor cell, a micro-crack that works as the electric by-pass is attractive. To make the by-pass, micro-processing by a femtosecond (fs) laser was applied We have achieved to hole the glass wall of the capillary tube with a few micrometer in diameter, however the processing has not completed yet because of lack of the precision for the laser beam focusing inside the small wall. Further trial is required to succeed in the production of the micro hole. When the pulsed light from the fs laser is irradiated to the biological tissue, second harmonic generation light (SHG light) is induced. Based on this phenomenon, we have constructed a polarization SHG light detection system. We found that orientation of the collagen fiber has been informed by the SHG light. As another trial of the cell sensor system, we have utilized a Paramecium caudatum as the sensor cell to monitor the water pollution based on the analysis of the cell moving profile in a micro-chamber. The paramecium cell showed characteristic moving profiles due to the pollutant such as heavy metal ions and acid. We have also studied the emission characteristics of a white light emitting diode to evaluate applicability to the compact light source in the present sensor system.
• 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

  Date (from‐to) : 2000/04 -2001/03 

  Author : 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

  Date (from‐to) : 1999/04 -2001/03 

  Author : 橋本 守
• Development of photometric detection system for tissue aging and study the localization of the aging in tissue

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2000 -2001 

  Author : ARAKI Tsutomu, TOHNO Yoshiyuki, YASUI Takeshi, HASHIMOTO Mamoru, YAMADA Gen

   

  The aim of this project is to develop an effective photometry system that can detect tissue aging of living body and to study the localization of tissue aging. The results of the project are described in the following. (1) A time-resolved confocal-microscope system with subnanosecond-resolution has been developed. Three-dimensional kinetic fluorescence image can be measured with this system. Image of calcium concentration in an yeast cell has been demonstraed based on the fluorescence lifetime image measurement. (2) Fluorescent collagen has been extracted from human dentin using acetic acid. The fluorescence spectra of the tooth-extracted collagen was consistent with that of the dentin tissue. Also the fluorescence intensity of the tooth-extracted collagen increased with the age, and with reaction time in the ribose solution. We have concluded that the main fluorophore of the age dependent fluorescence emission is AGE (Advanced Glycosylation Endproduct) that is generated in tissue collagen by the reaction with reducing sugar. (3) The second-harmonic generation light (SHG-light) detection system has been constructed. SHG-light is sensitive to tissue collagen conformation. We found that orientation of the collagen fiber has been informed by the SHG light. (4) Calcium content at bifurcation of the human brachial artery has been measured with an Ar-microwave-induced plasma emission spectrophotometer, and a possible correlation between Ca content and mechanical property of the artery wall has been considered. We have found that the portions in which Ca deposition is significant are the regions of low wall shear stress, and concluded that arteriosclerosis occurs in the regions of low wall share stress.
• 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：

  Date (from‐to) : 1999/11 -2000/10 

  Author : 橋本 守
• 細胞の持つ優れた分子選択能力を活用したバイオセンサー型クロマトグラフィの開発

  日本学術振興会：科学研究費助成事業

  Date (from‐to) : 1999 -2000 

  Author : 荒木 勉, 橋本 守

   

  細胞の環境感知能力と光学的手法による情報・収集技法を併用した生物マイクロセンサー方式のキャピラリー電気泳動システムの開発を目的としている。細胞に薬物や環境汚染物質などの刺激が加わるとミトコンドリア膜電位が変化したり、細胞質にカルシウムが放出されることに着目した。具体的には、細胞刺激物質の検出をCaを指標にして行う。センサー細胞として前年度には酵母菌を用い、Ca指示蛍光色素であるFluo-3で染色したが、キャピラリー両端にかかる電界(約150V/cm)によって蛍光強度が漸減するため、応用性に問題があった。そこで今年度は2波長励起による比測定法を採用した。そのための専用の顕微蛍光測光システムを製作した。センサー細胞には人の白血球を用いた。全血より白血球を分離し、Ca指示色素Fura-2で染色する。細胞を長さ31cm,内径25μmのガラス管(キャピラリー)内の中央付近に付着させ、キャピラリー両端に2-10kVの直流電圧をかける。はじめに蛍光強度の電界依存性を調査したが、印加電圧が4.5kVまでは影響がなかった。次にサンプル槽にアセチルコリンを注入すると、4.5kVの電圧ではアセチルコリンが約7分かかって細胞まで到達し、細胞を刺激する。これによって細胞内Caが上昇して、蛍光信号が得られた。その際、2波長情報からCa濃度が定量できた。さらにMgイオンについても、その検出のためにMag-fura-2の染色を試みた。以上の結果より、細胞をセンサーチップとした新しい電気泳動システムが実現可能であることがわかった。これらの結果を日本機械学会にて発表した。
• Detection of tissue aging by nanosecond self-fluorescence caused by AGE (Advanced Glycosylation Endproduct)

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 1999 -2000 

  Author : TSUTOMU Araki, GEN Yamada, YOSHIYUKI Tohno, MAMORU Hashimoto

   

  The aim of this project is to develop photometry system that can detect tissue aging of living body. The results of the project are described in the following. (1) The tissue fluoresces blue light by illumination with UV-light. The observed fluorescence decay time was in nanosecond range when illuminated with impulse UV-light. On the human dentine and human arterial tissue, the fluorescence intensity increased and fluorescence decay time decreased with increase of age, respectively. These results show that tissue fluorescence can be used as an effective indicator for aging. (2) The fluorescence decay curve had two decay components whose lifetimes are 2.8 ns and 9 ns. The intensity of 2.8 ns component showed strong age dependency, but 9 ns one showed weak dependency (3) We assumed that the main fluorophore of the age dependent fluorescence emission is AGE (Advanced Glycosylation Endproduct) that is generated in tissue collagen by the reaction with reducing sugar. To confirm this, type I collagen plate, human dentin section, rat tail tendon were incubated with ribose solution. Resultant fluorescence intensity and fluorescence decay time increased and decreased, respectively, with increase of the incubation time. The 2.8 ns component more increased than the 9 ns one. (4) To inform the change of collagen characteristics due to AGE, a new technique that can visualize the conformation change of molecules in living sample has been developed. This technique is based on the coherent anti-Stokes Raman spectroscopy (CARS). To establish the microscopic CARS method, coherent and optical transfer functions of the CARS system have been studied.
• Visualization and quantification of intracellular magnesium ion to determine the influence of magnesium to pragnancy circulation

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 1998 -1999 

  Author : ARAKI Tsutomu, HASHIMOTO Mamoru, YAMAZAKI Mineo, TOHNO Yoshiyuki

   

  One aim of this project is to develop a microphotometric measurement system that can visualize and quantify the intracellular magnesium ion level, and to apply this system into the study on the dynamics of magnesium ion during the pregnancy circulation. To visualize the intracellular magnesium ion, a fluorescent probe (Mag-fra2) is used in conjunction with a micro spectrophotometer that has two excitation wavelengths to take an intensity ratio of the fluorescence emission. Because the calcium ion level and the magnesium ion level is competetive in the cell, both calcium level and magnesium level must be measured with the same apparatus. The test apparatus has been completed. Relationship between the magnesium ion level in thrombocyte and the pregnancy has been investigated in preeclamptic pragnant women. The project has been extended to quantify the calcium in human blood vessels. The calcium is quantified by atomic emission spectroscoy (AES). For this, a microwave-induced coupled AES system has been made. The calcium content of lower limbs was higher than that of upper limbs. This result indicates that correlation between Ca content and arteriosclerosis maybe exists. Ca was much contained in parts of veinlet and joint in the artery comparing with other portion. This indicates that correlation between Ca content and mechanical stress such as exercise (living custom), blood pressure, and blood turbulent flow maybe exists. Also a new technique that can visualize the conformation change of molecules in living sample has been developed. This technique is based on the microscopic optics and coherent anti-Stokes Raman spectroscopy (CARS). To establish the microscopic CARS method, optical transfer function of the CARS system has been studied.

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