<i>Thermus thermophilus</i> polyploid cells directly imaged by X-ray laser diffraction
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- Suzuki Akihiro
- Research Institute for Electronic Science, Hokkaido University
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- Moriya Toshiyuki
- Institute of Environmental Microbiology, Kyowa-Kako Co., Ltd.
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- Oshima Tairo
- Institute of Environmental Microbiology, Kyowa-Kako Co., Ltd.
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- Yang Ying
- Research Institute for Electronic Science, Hokkaido University
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- Niida Yoshiya
- Research Institute for Electronic Science, Hokkaido University
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- Tono Kensuke
- RIKEN SPring-8 Center Japan Synchrotron Radiation Research Institute/SPring-8
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- Yabashi Makina
- RIKEN SPring-8 Center
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- Ishikawa Tetsuya
- RIKEN SPring-8 Center
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- Joti Yasumasa
- RIKEN SPring-8 Center Japan Synchrotron Radiation Research Institute/SPring-8
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- Nishino Yoshinori
- Research Institute for Electronic Science, Hokkaido University
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- Bessho Yoshitaka
- RIKEN SPring-8 Center Center for Interdisciplinary AI and Data Science, Ochanomizu University
Abstract
<p>Thermus thermophilus is reportedly polyploid and carries four to five identical genome copies per cell, based on molecular biological experiments. To directly detect polyploidy in this bacterium, we performed live cell imaging by X-ray free-electron laser (XFEL) diffraction and observed its internal structures. The use of femtosecond XFEL pulses enables snapshots of live, undamaged cells. For successful XFEL imaging, we developed a bacterial culture method using a starch- and casein-rich medium that produces a predominance of rod-shaped cells shorter than the focused XFEL beam size, which is slightly smaller than 2 µm. When cultured in the developed medium, the length of T. thermophilus cells, which is typically ~4 µm, was less than half its usual length. We placed living cells in a micro-liquid enclosure array and successively exposed each enclosure to a single XFEL pulse. A cell image was successfully obtained by the coherent diffractive imaging technique with iterative phase retrieval calculations. The reconstructed cell image revealed five peaks, which are most likely to be nucleoids, arranged in a row in the polyploid cell without gaps. This study demonstrates that XFELs offer a novel approach for visualizing the internal nanostructures of living, micrometer-sized, polyploid bacterial cells. </p>
Journal
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- The Journal of General and Applied Microbiology
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The Journal of General and Applied Microbiology 69 (2), 125-130, 2023
Applied Microbiology, Molecular and Cellular Biosciences Research Foundation