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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：89.93(Microscopy)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.
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.
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.
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
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.
