Thermus thermophilus polyploid cells directly imaged by X-ray laser diffraction.

IF 0.8 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of General and Applied Microbiology Pub Date : 2023-11-15 Epub Date: 2023-06-10 DOI:10.2323/jgam.2023.06.002
Akihiro Suzuki, Toshiyuki Moriya, Tairo Oshima, Ying Yang, Yoshiya Niida, Kensuke Tono, Makina Yabashi, Tetsuya Ishikawa, Yasumasa Joti, Yoshinori Nishino, Yoshitaka Bessho
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Abstract

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.

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用x射线激光衍射直接成像嗜热热菌多倍体细胞。
根据分子生物学实验,据报道,嗜热热菌是多倍体,每个细胞携带4到5个相同的基因组拷贝。为了直接检测该细菌的多倍体,我们利用x射线自由电子激光(XFEL)衍射进行了活细胞成像,并观察了其内部结构。飞秒XFEL脉冲的使用使活的、未受损细胞的快照成为可能。为了成功地进行XFEL成像,我们开发了一种细菌培养方法,使用富含淀粉和酪蛋白的培养基,产生比聚焦的XFEL光束尺寸短的棒状细胞,略小于2µm。当在成熟的培养基中培养时,嗜热t细胞的长度通常为~4µm,不到其通常长度的一半。我们将活细胞放置在微液体外壳阵列中,并连续地将每个外壳暴露在单个XFEL脉冲中。利用相干衍射成像技术,通过迭代相位恢复计算,成功地获得了细胞图像。重建的细胞图像显示了5个峰,最有可能是类核,在多倍体细胞中排列成一行,没有间隙。这项研究表明,XFELs为观察活的、微米大小的多倍体细菌细胞的内部纳米结构提供了一种新的方法。
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来源期刊
Journal of General and Applied Microbiology
Journal of General and Applied Microbiology 生物-生物工程与应用微生物
CiteScore
2.40
自引率
0.00%
发文量
42
审稿时长
6-12 weeks
期刊介绍: JGAM is going to publish scientific reports containing novel and significant microbiological findings, which are mainly devoted to the following categories: Antibiotics and Secondary Metabolites; Biotechnology and Metabolic Engineering; Developmental Microbiology; Environmental Microbiology and Bioremediation; Enzymology; Eukaryotic Microbiology; Evolution and Phylogenetics; Genome Integrity and Plasticity; Microalgae and Photosynthesis; Microbiology for Food; Molecular Genetics; Physiology and Cell Surface; Synthetic and Systems Microbiology.
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