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Quantitative visualization of stable isotope-labeled chromosome using isotope nanoscope
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| Title: | Quantitative visualization of stable isotope-labeled chromosome using isotope nanoscope |
| Other Titles: | 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 |
| Authors: | 永田, 康祐1 Browse this author |
| Authors(alt): | Nagata, Kosuke1 |
| Issue Date: | 25-Mar-2021 |
| Publisher: | Hokkaido University |
| Abstract: | 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. |
| Conffering University: | 北海道大学 |
| Degree Report Number: | 甲第14366号 |
| Degree Level: | 博士 |
| Degree Discipline: | 理学 |
| Examination Committee Members: | (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 |
| Degree Affiliation: | 理学院（自然史科学専攻） |
| Type: | theses (doctoral) |
| URI: | http://hdl.handle.net/2115/81966 |
| Appears in Collections: | 課程博士 (Doctorate by way of Advanced Course) > 理学院(Graduate School of Science)
学位論文 (Theses) > 博士 （理学）
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