同位体ナノスコープによる安定同位体標識された染色体の定量的な可視化
| Autor: | Nagata, Kosuke |
|---|---|
| Přispěvatelé: | 圦本, 尚義, 中川, 光弘, 川野, 潤, 馬上, 謙一 |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Popis: | Visualization of DNA strands and chromosomes have been used for the studies of DNA replication, resistance for mutagenesis and DNA repairing, and developed by using radio isotopes or analogs to label nucleotides throughout the cell cycles. However, these labels incorporated into DNA strand restrict the analysis of long-term dynamics of chromosomes because of its mutagenesis resulting in the mutations in cells. We applied stable isotopes for the nucleotide labeling to analyze spontaneous functions of chromosome. In this study, the imaging performance of isotope nanoscope and the consistence of the changes of the label amounts in the chromosomes were studied to evaluate the traceability of stable isotope labels. Firstly, the diameters of primary ion beam with aberration correction mode and non-correction mode were compared with the primary ion current to assess the imaging performance of isotope nanoscope. Aberration corrected primary ion beam showed smaller diameter and twice higher current density at a given lateral resolution. As a result, isotope nanoscope with aberration corrected primary ion beam indicated the abilities to produce fine and dense primary ion beam with lower acceleration voltage resulting in higher lateral resolutions and sensitivities and lower sample disruptions compared with conventional imaging mass spectrometers. Then, the chromosomes in human cultured cell labeled by U-13C6-Glucose throughout the cell cycles were visualized by isotope nanoscope. The heterogeneities of carbon isotope abundance were observed in each chromosome, and they reflected the semi-conservative replication of DNA strands. The distributions of stable isotope labels also suggested dispersive histone-incorporations into the chromatids. This nonmutagenic and subcellular resolution imaging may be useful to analyze natural functions of organelles with in single cells without radio isotope or analog usages. (主査) 教授 圦本 尚義, 教授 中川 光弘, 准教授 川野 潤, 助教 馬上 謙一 理学院（自然史科学専攻） |
| Databáze: | OpenAIRE |
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