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Journal Articles

Correlative imaging of live biological cells with a soft X-ray microscope and a fluorescence microscope

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

 Times Cited Count:3 Percentile:85.61

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.

Journal Articles

Nano-meter size modification of metal surfaces induced by soft X-ray laser single pulse

Ishino, Masahiko; Faenov, A.*; Tanaka, Momoko; Pikuz, T.; Tamotsu, Satoshi*; Hasegawa, Noboru; Nishikino, Masaharu; Starikov, S. V.*; Stegailov, V. V.*; Norman, G.*; et al.

X-Ray Lasers 2012; Springer Proceedings in Physics, Vol.147, p.121 - 124, 2014/00

 Times Cited Count:0 Percentile:0

Journal Articles

In situ observation of cellular organelles with a contact X-ray microscope

Kado, Masataka; Kishimoto, Maki; Tamotsu, Satoshi*; Yasuda, Keiko*; Shinohara, Kunio*

Journal of Physics; Conference Series, 463, p.012056_1 - 012056_4, 2013/10

 Times Cited Count:14 Percentile:96.95

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 1.3$$times$$10$$^{15}$$ 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.

Journal Articles

Nano-meter scale modifications on material surfaces induced by soft X-ray laser pulse irradiations

Ishino, Masahiko; Faenov, A.*; Tanaka, Momoko; Tamotsu, Satoshi*; Pikuz, T.; Hasegawa, Noboru; Nishikino, Masaharu; Inogamov, N.*; Skobelev, I.*; Fortov, V.*; et al.

Proceedings of SPIE, Vol.8849, p.88490F_1 - 88490F_8, 2013/09

 Times Cited Count:2 Percentile:73.17

Journal Articles

Imaging of fine structures of cellular organelles in hydrated biological cells by a soft X-ray microscope combined with a fluorescence microscope

Kado, Masataka; Kishimoto, Maki; Tamotsu, Satoshi*; Yasuda, Keiko*; Aoyama, Masato*; Shinohara, Kunio*

Proceedings of SPIE, Vol.8849, p.88490C_1 - 88490C_7, 2013/09

 Times Cited Count:1 Percentile:55.89

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.

Journal Articles

Observations of surface modifications induced by the multiple pulse irradiation using a soft picosecond X-ray laser beam

Ishino, Masahiko; Faenov, A. Ya.*; Tanaka, Momoko; Tamotsu, Satoshi*; Hasegawa, Noboru; Nishikino, Masaharu; Pikuz, T.; Kaihori, Takeshi*; Kawachi, Tetsuya

Applied Physics A, 110(1), p.179 - 188, 2013/01

 Times Cited Count:26 Percentile:71.46(Materials Science, Multidisciplinary)

Journal Articles

Interaction of soft X-ray laser pulse radiation with aluminum surface; Nano-meter size surface modification

Ishino, Masahiko; Faenov, A.*; Tanaka, Momoko; Hasegawa, Noboru; Nishikino, Masaharu; Tamotsu, Satoshi*; Pikuz, S.*; Inogamov, N. A.*; Zhakhovsky, V. V.*; Skobelev, I.*; et al.

AIP Conference Proceedings 1465, p.236 - 240, 2012/07

 Times Cited Count:2 Percentile:63.75

We irradiated the focusing soft X-ray laser pulses having a wavelength of 13.9 nm and the duration of 7 ps to aluminum surface. After the irradiation process, the irradiated surface was observed with a scanning electron microscope. The surface modifications caused by soft X-ray laser single pulse exposure were clearly seen. In addition, it was found that the conical structures having around 100 nm in diameters were formed in the shallow features. The nano-meter size modified surface structures on aluminum induced by the soft X-ray laser pulse is interesting as the newly surface structure. Hence, the soft X-ray laser beam would be a candidate for a tool of micromachining. We also provide a thermomechanical modeling of the soft X-ray laser interaction with aluminum briefly to explain the surface modification.

Journal Articles

Development of single shot soft X-ray contact microscopy system for nano-scale dynamics measurement of living biological specimen

Kishimoto, Maki; Kado, Masataka; Ishino, Masahiko; Tamotsu, Satoshi*; Yasuda, Keiko*; Shinohara, Kunio*

AIP Conference Proceedings 1465, p.43 - 47, 2012/07

 Times Cited Count:9 Percentile:94.57

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.

Journal Articles

Observation of organelle by a laser plasma X-ray microscope

Kado, Masataka; Kishimoto, Maki; Ishino, Masahiko; Tamotsu, Satoshi*; Yasuda, Keiko*; Shinohara, Kunio*

AIP Conference Proceedings 1465, p.246 - 250, 2012/07

 Times Cited Count:5 Percentile:85.25

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.

Journal Articles

Nanomodification of gold surface by picosecond soft X-ray laser pulse

Norman, G.*; Starikov, S.*; Stegailov, V.*; Fortov, V.*; Skobelev, I.*; Pikuz, T.; Faenov, A.*; Tamotsu, Satoshi*; Kato, Yoshiaki*; Ishino, Masahiko; et al.

Journal of Applied Physics, 112(1), p.013104_1 - 013104_8, 2012/07

 Times Cited Count:41 Percentile:82.44(Physics, Applied)

Journal Articles

Surface modifications of metals induced by soft X-ray laser pulse irradiations

Ishino, Masahiko; Faenov, A. Y.*; Tanaka, Momoko; Hasegawa, Noboru; Nishikino, Masaharu; Tamotsu, Satoshi*; Pikuz, T.; Oba, Toshiyuki*; Kaihori, Takeshi; Kawachi, Tetsuya

Journal of Laser Micro/Nanoengineering, 7(2), p.147 - 151, 2012/05

 Times Cited Count:5 Percentile:28.15(Nanoscience & Nanotechnology)

We irradiated the soft X-ray laser (SXRL) pulses having a wavelength of 13.9 nm, the duration time of 7 ps to aluminum (Al), copper (Cu) and gold (Au) surfaces. After the irradiation processes of SXRL beam with laser flux of $$sim$$14 mJ/cm$$^{2}$$ for Al case and of $$sim$$21 mJ/cm$$^{2}$$ for Cu and Au cases, the modified surfaces were observed with the visible microscope, the scanning electron microscope, and the atomic force microscope. The surface modifications caused by the SXRL irradiations were clearly seen on the surfaces, and it was found that the conical structures having around 100 nm in diameters were formed on the Al surface under a single pulse shot. The conical structures were formed in the features with the average depth of about 40 nm, and this value was in accordance with the attenuation length of the SXRL beam for Al. The modified structure on Al surface induced by SXRL pulse irradiations is different from those of Cu and Au surfaces. The modified structure formed on Al surface induced by the SXRL pulse exposure is interesting as the newly structure. Hence, the SXRL beam would be a candidate for a tool of micromachining, which enable to fabricate of three dimensional structures with nano-meter size on Al surface.

Journal Articles

Observation of organelles in Leydig cells by contact soft X-ray microscopy with a laser plasma X-ray source

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

 Times Cited Count:7 Percentile:89.95

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.

Journal Articles

Development of a specimen holder combined with ultra thin film laser plasma X-ray source for compact contact-type soft X-ray microscope to observe hydrated living biologocal cells

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

 Times Cited Count:0 Percentile:0.01

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.

Journal Articles

Flash imaging of fine structures of cellular organelles by contact X-ray microscopy with a high intensity laser plasma X-ray source

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.

Journal Articles

Laser ablation of gold; Experiment and atomistic simulation

Starikov, S. V.*; Stegailov, V. V.*; Norman, G. E.*; Fortov, V. E.*; Ishino, Masahiko; Tanaka, Momoko; Hasegawa, Noboru; Nishikino, Masaharu; Oba, Toshiyuki*; Kaihori, Takeshi; et al.

JETP Letters, 93(11), p.642 - 647, 2011/04

 Times Cited Count:25 Percentile:75.67(Physics, Multidisciplinary)

Journal Articles

Nanoscale surface modifications and formation of conical structures at aluminum surface induced by single shot exposure of soft X-ray laser pulse

Ishino, Masahiko; Faenov, A. Ya.*; Tanaka, Momoko; Hasegawa, Noboru; Nishikino, Masaharu; Tamotsu, Satoshi*; Pikuz, T. A.*; Inogamov, N. A.*; Zhakhovskii, V. V.*; Skobelev, I. Yu.*; et al.

Journal of Applied Physics, 109(1), p.013504_1 - 013504_6, 2011/01

 Times Cited Count:33 Percentile:77.1(Physics, Applied)

no abstracts in English

Journal Articles

Observation of actin filaments in Leydig cells with a contact-type soft X-ray microscope with laser plasma X-ray source

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.

Journal Articles

Observations of the intense soft X-ray emissions from ultra thin Au films irradiated with high contrast laser pulses

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

 Times Cited Count:2 Percentile:72.88

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$$^{5}$$ photons/$$mu$$m$$^{2}$$ 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.

Oral presentation

Observations of living cells by use of a soft X-ray microscope and direct comparison of cell images obtained with a fluorescence microscope and a soft X-ray microscope

Ishino, Masahiko; Kado, Masataka; Kishimoto, Maki; Nishikino, Masaharu; Hasegawa, Noboru; Oba, Toshiyuki; Kawachi, Tetsuya; Tamotsu, Satoshi*; Yasuda, Keiko*; Yamamoto, Yoshimasa*; et al.

no journal, , 

no abstracts in English

Oral presentation

Biological imaging by soft X ray, 2; Imaging of organelles

Kado, Masataka; Ishino, Masahiko; Kishimoto, Maki; Shinohara, Kunio*; Tamotsu, Satoshi*; Yasuda, Keiko*; Yamamoto, Yoshimasa*; Kinjo, Yasuhito*

no journal, , 

Contact soft X-ray microscopes with laser plasma X-ray sources are anticipated technology to be able to observe live hydrated biological cells with high spatial resolution. However there were no works reported which identified specific organelles clearly, because the obtained X-ray images are too complicated due to overlapping of cell structures. We have observed the same biological cells with both of a contact soft X-ray microscope and a confocal laser microscope and compared images obtained with both microscopes in order to identify organelles obtained with the soft X-ray microscope. As the results we succeeded to identify actin filaments and mitochondria clearly and found that the organelles obtained with the soft X-ray microscope were more detailed than those with the confocal laser microscope.

51 (Records 1-20 displayed on this page)