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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊東霖(T-Tony Yang) | |
| dc.contributor.author | Jia-Hui Wang | en |
| dc.contributor.author | 王嘉會 | zh_TW |
| dc.date.accessioned | 2022-11-24T03:38:20Z | - |
| dc.date.available | 2021-08-04 | |
| dc.date.available | 2022-11-24T03:38:20Z | - |
| dc.date.copyright | 2021-08-04 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-08-02 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81244 | - |
| dc.description.abstract | 單分子定位顯微鏡 (SMLM) 技術的最新發展使研究人員能夠在納米尺度上研究大分子結構。超分辨率技術的分辨率通過有效地銳化點擴散函數打破了光學顯微鏡的衍射極限。 單分子定位顯微鏡在眾多超解析方法中實現了十至二十納米的卓越分辨率，並從包含數十萬個定位的點雲數據集重建了不受到衍射限制的超解析影像。能夠對蛋白質進行共定位分析的多色光學顯微分析對研究者來說至關重要。儘管已經有許多用於多色成像的方法被提出，但其中大多數都需要多個激發波長和收光波段，通過連續採集或光譜分解。 目前尚未有技術可以在單一激發波長和收光波段，同時獲得多色的無色像差超解析影像。本研究提出了一種基於dSTORM（direct stochastic optical reconstruction microscopy）上的共時雙色成像策略，具有單個收光波段，可最效地減少色像差。本研究闡述了 Alexa fluor 647 和 ATTO647N 的光動力學，並開發了一種數據驅動的螢光染料分辨方法。此外，這種方法可以被應用於任何 dSTORM 成像系統，而無需額外的光學組件來分離重疊光譜內的螢光團。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T03:38:20Z (GMT). No. of bitstreams: 1 U0001-2807202111000600.pdf: 3592982 bytes, checksum: b3070c0426ba98d7d04ab2e4f0c6bc1e (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | CONTENTS 審定書 i 致謝 ii 中文摘要 iii ABSTRACT iv CONTENTS v LIST OF FIGURES viii LIST OF TABLES xi Chapter 1 Introduction 1 1.1 Background 1 1.2 Literature review 2 1.2.1 Resolution limitation of optical microscopy 2 1.2.2 Superresolution methods 3 1.2.3 dSTORM superresolution technique 5 1.2.4 Multicolor SMLM imaging 7 1.3 Research motivation 9 1.4 Thesis structure 11 Chapter 2 Experimental Design 12 2.1 Characterization of far-red dye candidates 12 2.1.1 Evaluation of fluorophore candidates 12 2.1.2 AF647 and ATTO647N as fluorophore candidates 14 2.2 Quantification of photo-switching kinetics of fluorophores 19 2.2.1 Switching kinetics of AF647 and ATTO647N 19 2.2.2 Fast Fourier transfer of Switching kinetics 25 Chapter 3 Materials and Methods 30 3.1 Cell culture 30 3.2 Immunostaining 30 3.3 dSTORM imaging 32 3.4 Data analysis 34 Chapter 4 Results and Discussions 38 4.1 Validation of the chromatic aberration-free two-color imaging 38 4.1.1 Fluorophore separation of AF647 and ATTO647N 38 4.1.2 Separation method based on support vector machine (SVM) 40 4.2 Optimization of fluorophore separation 45 4.2.1 On-state and off-state analysis for color separation 45 4.2.2 Separation method based on ensemble forest method 48 4.3 Superresolution imaging of cellular structure 56 4.3.1 dSTORM imaging of Arl13b and Nup98 56 4.3.2 dSTORM imaging of FBF1 and ARL13b 58 4.3.3 dSTORM imaging of FBF1 and Ninein 60 4.4 Discussion 61 Chapter 5 Conclusion 65 Chapter 6 Future work 67 6.1 Multicolor imaging 67 6.2 Image quality improvement 68 Reference 69 | |
| dc.language.iso | en | |
| dc.subject | 多色成像 | zh_TW |
| dc.subject | dSTORM | zh_TW |
| dc.subject | 超分辨率 | zh_TW |
| dc.subject | 共時 | zh_TW |
| dc.subject | 無像差 | zh_TW |
| dc.subject | simultaneous | en |
| dc.subject | dSTORM | en |
| dc.subject | multicolor imaging | en |
| dc.subject | aberration free | en |
| dc.subject | superresolution | en |
| dc.title | 共時無色相差雙色單分子定位顯微技術之設計與實驗 | zh_TW |
| dc.title | Design and Experiment for the Chromatic Aberration Free Two-Color Single-Molecule Localization Microscopy | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 黃念祖(Hsin-Tsai Liu),陳奕帆(Chih-Yang Tseng) | |
| dc.subject.keyword | dSTORM,超分辨率,共時,無像差,多色成像, | zh_TW |
| dc.subject.keyword | dSTORM,superresolution,simultaneous,aberration free,multicolor imaging, | en |
| dc.relation.page | 83 | |
| dc.identifier.doi | 10.6342/NTU202101837 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-08-02 | |
| dc.contributor.author-college | 電機資訊學院 | zh_TW |
| dc.contributor.author-dept | 生醫電子與資訊學研究所 | zh_TW |
| 顯示於系所單位： | 生醫電子與資訊學研究所 | |
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