以超解析影像技術觀測端粒長度

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Title:	以超解析影像技術觀測端粒長度
Other Titles:	Observation of telomere lengths by super-resolution imaging technology
Authors:	林蔚青 LIN, WEI-CHING
Contributors:	張柏齡 CHANG, PO-LING 化學系
Keywords:	端粒長度;超解析螢光顯微鏡 Telomere length;Super-resolution fluorescence microscopy
Date:	2019
Issue Date:	2019-12-16T02:21:27Z (UTC)
Abstract:	端粒（telomere）是位於染色體末端由重複核酸序列組合而成的結構，用於維持染色體的完整性，而端粒的長度會隨著細胞分裂或是環境壓力而逐漸縮短，研究指出端粒長度的變化是導致人體衰老及癌症產生的重要指針，因此端粒長度變化的測量是癌症預防的重要手段。近年來報導了許多方法來測量端粒長度，例如末端限制性片段分析（terminal restriction fragmentation，TRF），定量即時聚合酶連鎖反應（quantitative real-time polymerase chain reaction，Q-PCR），定量螢光原位雜合（quantitative fluorescence in situ hybridization，Q-FISH）及流式細胞螢光原位雜合（flow cytometry and flow fluorescence in situ hybridization，Flow-FISH）等。這些方法都有各自的優點，但卻有一個共通的弊病──無法測量線性化端粒DNA 的絕對長度，因此本實驗發展利用超解析影像技術來觀測端粒的絕對長度。目前我們已經成功將端粒DNA 線性沉積在疏水性玻片表面，透過端粒螢光標記探針定位測量端粒絕對長度，同時我們也成功分離出單顆細胞DNA 並嘗試將其完整的線性沉積在玻片上，希望未來能夠在單顆細胞的水平上測量出細胞端粒長度。 Telomere is the structure composed by repeating nucleic acid sequences at the end of a chromosome, and which to maintain the integrity of the chromosome. The length of the telomere is gradually shortened when the cell divides or the environmental pressure. The study shows that the change of the telomere length is an important indicator of human aging and cancer. Therefore, the measurement of telomere length changes is crucial means of cancer prevention. In recent years, a lot of methods have been reported to measure the length of telomere, such as terminal restriction fragmentation (TRF), quantitative real-time polymerase chain reaction (QPCR), quantitative fluorescein Quantitative fluorescence in situ hybridization (Q-FISH) and flow cytometry and flow fluorescence in situ hybridization (Flow-FISH). All above the mentions, these methods have their own advantages, however, there is a common drawback—the absolute length of linearized telomere DNA cannot be measured. Hence, this experiment develops the techniques of super-resolution image to measure the absolute length of telomeres. We have successfully deposited telomere DNA on the surface of hydrophobic slides and measured the absolute length of telomeres through telomere fluorescent labeled probes at the present time. Simultaneously, we also achieved to isolate the DNA of a single cell and tried to completely deposit it on slides. It is hope that we can measure the telomere lengths at the level of single cell.
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