Kado, Masataka; Kishimoto, Maki; Tamotsu, Satoshi*; Yasuda, Keiko*; Aoyama, Masato*; Tone, Shigenobu*; Shinohara, Kunio*
AIP Conference Proceedings 1696, p.020019_1 - 020019_4, 2016/01
Soft X-ray microscope is a very powerful tool to observe cellular organelles of living biological cells and many works have demonstrated imaging of inner structures of the cells. However the inner structures are very complicated and it is difficult to identify the organelles obtained with the soft X-ray microscopes. We have proposed a hybrid imaging method with a soft X-ray microscope and a fluorescence microscope that is to observe the same biological cells with the both microscopes at the same time. Using the information of the cellular organelles obtained with the fluorescence microscope, inner structures obtained with the soft X-ray microscope are accurately identified. We have observed living biological cells by the hybrid imaging method. Since the soft X-ray microscope has higher spatial resolution than that of the fluorescence microscope, fine structures of the cellular organelles in the living biological cells were discussed.
Namba, Shinichi*; Hasegawa, Noboru; Kishimoto, Maki; Nishikino, Masaharu; Ishino, Masahiko; Kawachi, Tetsuya
AIP Advances (Internet), 5(11), p.117101_1 - 117101_10, 2015/11
To characterize the temporal evolution of ultrashort X-ray pulses emitted by laser plasmas using a pump-probe method, a magnetic bottle time-of-flight electron spectrometer is constructed. The design is determined by numerical calculations of a mirror magnetic field and of the electron trajectory in a flight tube. The performance of the spectrometer is characterized by measuring the electron spectra of xenon atoms irradiated with a laser-driven plasma X-ray pulse. In addition, as a preliminary experiment, two-color above-threshold ionization (ATI) is conducted, in which xenon atoms are simultaneously irradiated with an X-ray laser pump and an IR laser probe. The correlation in the intensity of the Auger sideband spectra and in the time delay of the two laser pulses yields an X-ray pulse width of 5.7 ps, in good agreement with the value obtained using an X-ray streak camera.
Kado, Masataka; Kishimoto, Maki; Tamotsu, Satoshi*; Yasuda, Keiko*; Shinohara, Kunio*
Journal of Physics; Conference Series, 463, p.012056_1 - 012056_4, 2013/10
A contact X-ray microscope coupled with a high intense laser plasma soft X-ray source has been developed and in situ observations of cellular organelles have been conducted. The soft X-ray source were generated by a high power laser pulse onto a thin foiled gold target at the photon numbers of 1.310 photons/sr to be able to capture an image of live wet biological cells. The cells were cultured on PMMA photoresists that were formed on transparent glass plates to make optical microscope observation possible. The cells were observed by both of optical microscope and soft X-ray microscope. The obtained soft X-ray images were directly compared with corresponding fluorescent optical images. Cellular organelles such as mitochondria and cytoskeleton in the soft X-ray images were identified referencing the information obtained from the fluorescent images.
Kado, Masataka; Kishimoto, Maki; Tamotsu, Satoshi*; Yasuda, Keiko*; Aoyama, Masato*; Shinohara, Kunio*
Proceedings of SPIE, Vol.8849, p.88490C_1 - 88490C_7, 2013/09
We have proposed to use a fluorescence microscope to identify the cellular organelles in the images obtained with the soft X-ray microscope observing the same cells with both microscopes. The cells were stained with several fluorescent dyes such as Mito-tracker, Phalloidin, and DAPI and after taking many fluorescence images of cellular organelles the cells were exposed to the flash soft X-rays. The obtained soft X-ray images and fluorescence images of the cells were directly compared and each of the cellular organelles such as mitochondria, actin filaments, and chromosomes in the soft X-ray images was clearly identified. Since the soft X-ray microscope has higher spatial resolution than that of the fluorescence microscope, fine structures of the cellular organelles in the hydrated biological cells were observed for the first time.
Kishimoto, Maki; Kado, Masataka; Ishino, Masahiko; Tamotsu, Satoshi*; Yasuda, Keiko*; Shinohara, Kunio*
AIP Conference Proceedings 1465, p.43 - 47, 2012/07
We have been developing a picosecond single shot soft X-ray contact microscopy system for observing the nanometer-scale inner structure of the living biological specimen in a hydrated condition. The microscopy system consists of an intense IR pump laser system for generating soft X-rays and X-ray microscope chamber. The pump laser system employs OPCPA technique to generate water-window X-rays effectively. The X-ray microscope chamber is composed of a vacuum chamber, a focusing lens, a metal film target, an in-vacuum type sample holder. The soft X-rays from the laser-induced plasma generated by pump laser pulse illuminates bio-specimens on the PMMA photo resist set in the in-vacuum sample holder. The photo resist is developed and the X-ray transmission imageis read out by AFM. We took X-ray images of hydrated Leydig cells from mouse testicle and demonstrated that the developed X-ray microscopy system has a spatial resolution of about 100 nm.
Kado, Masataka; Kishimoto, Maki; Ishino, Masahiko; Tamotsu, Satoshi*; Yasuda, Keiko*; Shinohara, Kunio*
AIP Conference Proceedings 1465, p.246 - 250, 2012/07
Contact X-ray microscopy has a potential to image wet biological specimens in natural condition. It is very important to identify obtained features in the X-ray images, since X-ray microscopes have potential to image features that have not been visualized yet. We have proposed to compare the X-ray images of the biological specimens with the fluorescence images and to identify the features found in the X-ray images based on the features found in the fluorescence images. Comparing the X-ray images to the fluorescence images of the set biological cells, fine structures of the mitochondria in the X-ray images have been able to be identified.
Kawachi, Tetsuya; Hasegawa, Noboru; Nishikino, Masaharu; Ishino, Masahiko; Imazono, Takashi; Oba, Toshiyuki; Kaihori, Takeshi; Kishimoto, Maki; Ochi, Yoshihiro; Tanaka, Momoko; et al.
X-Ray Lasers 2010; Springer Proceedings in Physics, Vol.136, p.15 - 24, 2011/12
This paper reviews recent improvement in the source development of laser-driven X-ray lasers and the applications in the research fields of material science, laser processing, X-ray imaging, and radiation damage in biological cells. In the application for material science, we have firstly observed temporal correlation between the domain structures of ferro-electric substance under the Curie temperature. In the laser processing, new X-ray laser interferometer reveals us the nano-scale surface distortion of substance pumped by a femto-second optical pulse. In the X-ray diffraction image, we have taken several static images of micro-structure of samples: now we are trying to extend the objective to nano-scale dynamics using pump and probe method. In the radiation damage of biological cells, we observed double strand break in DNA using X-ray laser exposure; this results are compared with the case using incoherent several KeV X-ray exposure.
Hasegawa, Noboru; Ochi, Yoshihiro; Kawachi, Tetsuya; Terakawa, Kota*; Tomita, Takuro*; Yamamoto, Minoru; Nishikino, Masaharu; Oba, Toshiyuki; Kaihori, Takeshi; Imazono, Takashi; et al.
X-Ray Lasers 2010; Springer Proceedings in Physics, Vol.136, p.353 - 358, 2011/12
The understanding of the dynamics of the initial process is important for the micro processing and welding by the ultra-short laser pulse. The X-ray laser is suitable for probing this initial process because it has short wavelength (Ni-like Ag, 13.9 nm) and short duration (7 ps). For this investigation, the origin of time of the pumping pulse is quite important. In this study, we used the scintillation plate and the plasma gate technique to realize the spatial and temporal synchronization of the pump and probe pulses. For the spatial alignment, a CsI scintillation plate that was set at the sample position was illuminated by both the X-ray laser pulse, and the fluorescence light were detected by the CCD camera. For the temporal synchronization, we set a thin foil at the sample position. We measured the transmission of the X-ray laser while changing a temporal delay of the pumping laser with respect to the time of X-ray laser pulse to obtain the origin of the irradiation time.
Kado, Masataka; Ishino, Masahiko; Tamotsu, Satoshi*; Yasuda, Keiko*; Kishimoto, Maki; Nishikino, Masaharu; Kinjo, Yasuhito*; Shinohara, Kunio*
AIP Conference Proceedings 1365, p.391 - 394, 2011/09
Contact X-ray microscopy has achieved a single-shot imaging of wet biological specimens in natural condition and succeeded in imaging live mouse macrophages with hair-like structures, which was not observed before. It is very important to identify obtained features in the X-ray images, since X-ray microscopes have potential to image features that have not been visualized yet. Here, we demonstrate to image the same biological specimens both by confocal laser microscopy and soft X-ray microscopy. Staining biological specimens with well-established techniques makes easy to identify features in the fluorescence images obtained with confocal laser microscope. Comparing the X-ray images of the specimens with the fluorescence images, features found in the fluorescence images could also be identified in the X-ray images. Comparing the X-ray images to the fluorescence images, fine structures of the actin filaments in the X-ray images were identified.
Ishino, Masahiko; Kado, Masataka; Shinohara, Kunio*; Yamamoto, Yoshimasa*; Hirai, Itaru*; Kishimoto, Maki; Nishikino, Masaharu; Hasegawa, Noboru; Tamotsu, Satoshi*; Yasuda, Keiko*; et al.
Proceedings of SPIE Europe Optics + Optoelectronics 2011, Vol.8139, p.81390R_1 - 81390R_8, 2011/09
Ultra thin gold films are favorable laser plasma targets for a soft X-ray microscopy, because the thin films emit intense soft X-rays at the wavelength of water window region. Using rear side emissions, the distance between the X-ray source and the specimens can be reduced so that the X-ray flux on specimens increases. The microscope system can be designed to be compact when the specimen holder and X-ray source are combined in one piece. The biological specimen holder combined with an ultra thin film target has been developed. This X-ray microscope system needs not any X-ray optics which causes a decrease in X-ray photons for imaging. X-ray images of hydrated living cells have been obtained successfully by use of the newly developed specimen holder. Specimen holder combined with plasma X-ray source will be a key component of a compact soft X-ray microscope using in a laboratory.
Kado, Masataka; Ishino, Masahiko; Kishimoto, Maki; Tamotsu, Satoshi*; Yasuda, Keiko*; Kinjo, Yasuhito*; Shinohara, Kunio*
Proceedings of SPIE Europe Optics + Optoelectronics 2011, Vol.8139, p.81390O_1 - 81390O_7, 2011/09
Laser plasma X-ray sources have high intensity and short pulse duration, and are suitable for X-ray microscopy in biology. They make wet live biological specimens possible to be imaged with a single shot X-ray exposure and several works have been done to image them. However there were no reports on the imaging of fine structures of cellular organelles in a live biological cell since higher X-ray intensity is needed for it. We have developed a high intensity laser plasma X-ray source, cooperating it with contact X-ray microscopy, and observed fine structures of cellular organelles in a wet biological cells. Comparing the X-ray images and the fluorescence images of cellular organelles such as actin filaments and mitochondria we have been clearly able to identify organelles in the X-ray images and observed fine structures.
Inogamov, N. A.*; Anisimov, S. I.*; Petrov, Y. V.*; Khokhlov, V. A.*; Zhakhovskii, V. V.*; Faenov, A. Ya.*; Pikuz, T.; Fortov, V. E.*; Skobelev, I. Y.*; Kato, Yoshiaki*; et al.
Journal of Optical Technology, 78(8), p.473 - 480, 2011/08
Inogamov, N. A.*; Zhakhovsky, V. V.*; Faenov, A. Ya.*; Khokhlov, V. A.*; Shepelev, V. V.*; Skobelev, I. Y.*; Kato, Yoshiaki*; Tanaka, Momoko; Pikuz, T. A.*; Kishimoto, Maki; et al.
Applied Physics A, 101(1), p.87 - 96, 2010/10
Kado, Masataka; Ishino, Masahiko; Tamotsu, Satoshi*; Yasuda, Keiko*; Kishimoto, Maki; Nishikino, Masaharu; Kinjo, Yasuhito*; Shinohara, Kunio*
Denki Gakkai Rombunshi, C, 130(10), p.1774 - 1778, 2010/10
Actin filaments in Leydig cells from mouse testes have been observed with a contact-type soft X-ray microscope with laser plasma X-ray source. The Leydig cells were fixed with paraformaldehyde, stained with Phalloidin, and observed with a confocal laser microscope prior to the observation with X-ray microscope. Obtained images by both of the confocal laser microscopy and the X-ray microscopy were directly compared and revealed that not only position of actin filaments but also the shapes can be identified each other. The actin filaments in the X-ray images were clearly recognized and their structures were obtained in more detail compared to those in the confocal laser microscope images.
Inogamov, N. A.*; Faenov, A. Ya.*; Zhakhovskii, V. V.*; Skobelev, I. Y.*; Khokhlov, V. A.*; Kato, Yoshiaki*; Tanaka, Momoko; Pikuz, T. A.*; Kishimoto, Maki; Ishino, Masahiko; et al.
Contributions to Plasma Physics, 51(4), p.361 - 366, 2010/05
Ishino, Masahiko; Kado, Masataka; Nishikino, Masaharu; Shinohara, Kunio*; Tamotsu, Satoshi*; Yasuda, Keiko*; Hasegawa, Noboru; Kishimoto, Maki; Oba, Toshiyuki; Kawachi, Tetsuya
Proceedings of SPIE, Vol.7589, p.75891B_1 - 75891B_8, 2010/02
Soft X-ray microscopes operating in the water window are capable of imaging living hydrated biological specimens. Laser produced plasmas are attractive soft X-ray sources, because of their short duration time. Based on the minimum dose calculation, soft X-ray photons more than 10 photons/m at the sample surface are needed to acquire an image of the biological specimens with spatial resolution up to 100 nm. The observations of soft X-ray emissions from laser produced plasmas using ultra thin film targets have been carried out. Au thin films were irradiated by a high contrast Nd:glass laser pulses. The spectral properties of emitted soft X-rays were monitored by an X-ray spectrograph from the rear side with respect to the surface of laser irradiation. The observed emission intensities had an obvious dependence on the film thickness, and the most intense emissions were obtained at the thickness of 28 nm. The experimental results have suggested that the most of the laser energy irradiated is absorbed by the film target, and it is resulted an efficient energy deposition from laser to X-rays.
Kishimoto, Maki; Namikawa, Kazumichi*; Sukegawa, Kota*; Yamatani, Hiroshi*; Hasegawa, Noboru; Tanaka, Momoko
Review of Scientific Instruments, 81(1), p.013905_1 - 013905_5, 2010/01
We developed a new soft X-ray speckle intensity correlation spectroscopy system by use of a single shot high brilliant plasma soft X-ray laser. The plasma soft X-ray laser is characterized by several picoseconds in pulse width, more than 90% special coherence, and 10 soft X-ray photons within a single pulse. We developed a Michelson type delay pulse generator using a soft X-ray beam splitter to measure the intensity correlation of X-ray speckles from materials and succeeded in generating double coherent X-ray pulses with picosecond delay times. Moreover, we employed a high-speed soft X-ray streak camera for the picosecond time-resolved measurement of X-ray speckles. We performed the X-ray speckle intensity correlation measurements for probing the relaxation phenomena of polarizations in polarization clusters in the paraelectric phase of the ferroelectric material BaTiO near its Curie temperature and verified its performance.
Ochi, Yoshihiro; Kawachi, Tetsuya; Hasegawa, Noboru; Nishikino, Masaharu; Oba, Toshiyuki; Tanaka, Momoko; Kishimoto, Maki; Kaihori, Takeshi; Nagashima, Keisuke; Sugiyama, Akira
Japanese Journal of Applied Physics, 48(12), p.120212_1 - 120212_3, 2009/12
We have demonstrated the fully spatial coherent 13.9-nm laser with output energy of submicroJoule by means of the double target geometry by using new driver system. The power amplifiers providing tow beams with 10-J energy and picoseconds duration at the repetition rate of 0.1 Hz. The obtained 13.9-nm laser pulse with 10 coherent photons will enable us to make single-shot pump-probe observation of non-periodic ultrafast phenomena.
Faenov, A. Y.; Inogamov, N. A.*; Zhakhovskii, V. V.*; Khokhlov, V. A.*; Nishihara, Katsunobu*; Kato, Yoshiaki*; Tanaka, Momoko; Pikuz, T. A.*; Kishimoto, Maki; Ishino, Masahiko; et al.
Applied Physics Letters, 94(23), p.231107_1 - 231107_3, 2009/06
Hasegawa, Noboru; Kawachi, Tetsuya; Sasaki, Akira; Yamatani, Hiroshi; Kishimoto, Maki; Ochi, Yoshihiro; Tanaka, Momoko; Nishikino, Masaharu; Kunieda, Yuichi; Nagashima, Keisuke; et al.
Journal of Physics; Conference Series, 163(1), p.012062_1 - 012062_4, 2009/06
The precise knowledge about the wavelength and the spectral width of the lasing line is important for the applications of X-ray lasers, and especially for the spectral width, it is good benchmark of the atomic code because it depends on the electron collisional excitation and de-excitation rate coefficient. Only a few measurements of the spectral width of the laser line have been reported, because the spectral width of the X-ray laser is so narrow that the required spectral resolution is quite high. In this study, we took the nickel-like molybdenum X-ray laser as an example, and measure the spectral width by use of the high resolution spectrometer in order to compare it with a theoretical one.