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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 董成淵 | zh_TW |
| dc.contributor.advisor | Chen-Yuan Dong | en |
| dc.contributor.author | 黃品毓 | zh_TW |
| dc.contributor.author | Pin-Yu Huang | en |
| dc.date.accessioned | 2025-08-18T16:06:28Z | - |
| dc.date.available | 2025-08-19 | - |
| dc.date.copyright | 2025-08-18 | - |
| dc.date.issued | 2025 | - |
| dc.date.submitted | 2025-08-06 | - |
| dc.identifier.citation | [1] 衛生福利部統計處, "民國112年死因統計結果摘要表," 2023.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/98689 | - |
| dc.description.abstract | 頭頸癌為世界第七大癌症，每年造成全球四十餘萬人死亡。傳統療法對於末期的病患的成效有限，五年存活率僅約40%，而近期免疫療法的進展為癌症治療帶來一線曙光。免疫檢查點抑制劑 (immune checkpoint inhibitor, ICI) 為常用於頭頸癌的免疫療法藥物，透過阻斷免疫細胞表面的免疫檢查點與癌細胞結合，達到刺激免疫系統清除病患體內腫瘤的效果。然而，並非所有病患對免疫療法皆有反應。在預測病患的投藥後反應方面，當前以PD-L1做為生物指標的檢測方式僅有20~30% 的準確率。因此，尋找能有效預測病患反應的生物指標是當前免疫療法研究的重要課題。
在本研究中，我們試圖從腫瘤微環境(tumor microenvironment, TME)中免疫細胞的表現量尋找潛在的生物指標。從接受免疫療法的頭頸癌病患中，我們取各五位反應者與無反應者在投藥前的腫瘤石蠟切片作為樣品，以多重免疫螢光染色對組織內的6種免疫細胞、9種潛在生物指標進行標記，進而比較兩組病患細胞表現量的差異。我們搭建了四頻道的螢光顯微系統，對樣品進行全切片掃描(Whole Slide Imaging)，並以影像計算細胞數量，分析各種細胞在腫瘤內部的占比。 根據10組樣品的結果顯示，腫瘤內CD8+ T 細胞的比例在兩組病患間有顯著差異，與過往研究的結果相同，而腫瘤相關巨噬細胞 (TAM) 則無如過往研究呈現相關性。 本研究使用的系統尚須許多改進以滿足當前臨床研究的需求，未來將試圖增加頻道數、減少曝光時間以加快篩檢速度。 | zh_TW |
| dc.description.abstract | Head and neck squamous cell carcinoma (HNSCC) is the seventh most common cancer in the world. Due to poor prognosis, it has become the serious health issue in Taiwan. Recently, the progress in the immunotherapy shows effectiveness to the treatment of HNSCC. Patients receiving immune check point inhibitor (ICI) can be treated. However, only around 20 to 30% of the patients respond to the treatment. Therefore, searching for biomarkers that better predict the efficacy of immunotherapy has become an important issue is current research.
In this research, we established a four-channel multiplex immunofluorescence microscope to search potential biomarker in the tumor microenvironment (TME). Formalin-fixed, paraffin-embedded samples from 5 responders and 5 non-responders of HNSCC patients were studied. 9 markers were labeled in 3 cycles, and whole slide imaging was performed on 10 samples. The number of six types of immune cells within tumor was calculated through image cytometry. The intratumoral CD8+ T cell ratio shows a significant difference between two groups, which in accordance the result in previous research. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2025-08-18T16:06:28Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2025-08-18T16:06:28Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 誌謝 ii
摘要 iii Abstract iv CONTENTS v LIST OF FIGURES vii LIST OF TABLES viii Chapter 1 Introduction 1 1.1 Health Significance of Head and Neck Squamous Cell Carcinoma (HNSCC) 1 1.2 Current Treatment of HNSCC in the Clinics 1 1.3 Promise and Challenges of Immunotherapy in HNSCC Treatment 2 1.4 Immunotherapy Biomarker Search by Multiplex Immunofluorescence Microscopy 3 1.5 Biomarker Considerations for HNSCC Immunotherapy 4 Chapter 2 Principles of Multiplexed Immuno-fluorescence Microscopy (MIFM) 6 2.1 Fluorescence for Biomarker Detection 6 2.1.1 Advantages of Fluorescence Detection in Biomarker Search 6 2.1.2 Molecular and Optical Characteristics of Fluorophores 6 2.2 Resolution and Magnification Considerations in MIFM 8 2.2.1 Point Spread Function and Required Resolution 8 2.2.2 Magnification and Field of View 14 2.3 Multiplex Immunofluorescence Microscopy 15 2.3.1 Chemistry of Cyclic Labeling and Bleaching 15 2.3.2 Choice of Bleaching Agent 16 Chapter 3 Materials and Methods 17 3.1 Design of Wide Field Fluorescence Microscope (WFFM) 17 3.1.1 Excitation Source and Filters 18 3.1.2 Optical Path Design 19 3.1.3 Image Stitching for Large Area Imaging 20 3.2 Preparation of HNSCC Tissue Sections 23 3.2.1 Consideration of HNSCC Patient Specimens 23 3.2.2 Sectioning and Deparaffinization 23 3.2.3 Tissue Attachment and Preprocessing 24 3.2.4 Cyclic Labeling, Imaging and Bleaching 24 Chapter 4 Results and Discussion 26 4.1 Calibration of Wide Field Fluorescence Microscope 26 4.1.1 Estimate of Fluorescence Detection Efficiency 26 4.1.2 Field Flatness Correction 27 4.1.3 Field Size Calibration 29 4.1.4 Imaging of Fluorescent Microspheres in the Four-Channel System 29 4.1.5 Autofluorescence Removal of Tissue Sections 30 4.2 Multiplex Immunofluorescent Landscape in HNSCC Patients 32 4.2.1 Immunofluorescent Biomarkers Distribution in Responding Patients 32 4.2.2 Immunofluorescent Biomarkers Distribution in Non-Responding Patients 34 4.2.3 Comparison of Intratumoral immune cell ratio between responders and non-responders 35 Chapter 5 Conclusion 37 5.1 Future work 37 Appendix 39 Reference 56 | - |
| dc.language.iso | en | - |
| dc.subject | 頭頸癌 | zh_TW |
| dc.subject | 免疫療法 | zh_TW |
| dc.subject | 生物指標 | zh_TW |
| dc.subject | 多重免疫螢光染色 | zh_TW |
| dc.subject | 全切片掃描 | zh_TW |
| dc.subject | 腫瘤微環境 | zh_TW |
| dc.subject | biomarkers | en |
| dc.subject | Head and neck squamous cell carcinoma (HNSCC) | en |
| dc.subject | tumor microenvironment | en |
| dc.subject | whole slide imaging | en |
| dc.subject | cyclic multiplexed immunofluorescence microscopy | en |
| dc.subject | immunotherapy | en |
| dc.title | 免疫螢光顯微術用於癌症免疫療法之生物指標研究 | zh_TW |
| dc.title | Multiplex Immunofluorescence Microscopy for Biomarker Search in Cancer Immunotherapy | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 113-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 婁培人;陳永芳;朱士維 | zh_TW |
| dc.contributor.oralexamcommittee | Pei-Jen Lou;Yang-Fang Chen;Shi-Wei Chu | en |
| dc.subject.keyword | 頭頸癌,免疫療法,生物指標,多重免疫螢光染色,全切片掃描,腫瘤微環境, | zh_TW |
| dc.subject.keyword | Head and neck squamous cell carcinoma (HNSCC),immunotherapy,biomarkers,cyclic multiplexed immunofluorescence microscopy,whole slide imaging,tumor microenvironment, | en |
| dc.relation.page | 59 | - |
| dc.identifier.doi | 10.6342/NTU202502886 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2025-08-12 | - |
| dc.contributor.author-college | 理學院 | - |
| dc.contributor.author-dept | 應用物理研究所 | - |
| dc.date.embargo-lift | N/A | - |
| 顯示於系所單位： | 應用物理研究所 | |
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