嵌入式繼光鏡顯微超頻譜影像系統應用在口腔癌切片及活體之設計及研究

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謝耀方(Yao-Fang Hsieh) 查詢紙本館藏

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光電科學與工程學系

論文名稱

嵌入式繼光鏡顯微超頻譜影像系統應用在口腔癌切片及活體之設計及研究
(Research and Design of Finite Conjugate Embedded Relay Lens Microscopic Hyperspectral Imaging System (ERL-MHIS) on Oral Cancer in-vitro and in-vivo Application)

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摘要(中)

口腔癌的五年存活率約57%，若能早期發現，則能提升到80%左右。目前診斷口腔癌的方式仍以切片為主，但切片的判斷會有人為經驗上的誤差，因此病理醫生希望能找出量化診斷的儀器來輔助診斷口腔癌。由於顯微超頻譜影像系統能同時提供切片的組織形態資訊以及光譜資訊，近來被使用在in-vitro口腔癌的檢測，但因其掃描結構複雜且高倍率影像品質不佳，影響診斷準確率。因此本研究發展一套嵌入式繼光鏡超頻譜影像系統，並將其結合顯微鏡成為嵌入式繼光鏡顯微超頻譜影像系統，利用繼光鏡的遠心特性，將本來在顯微鏡物平面的掃描機制轉移到像平面，使用繼光鏡來掃描影像，如此掃描解析度不用隨著放大倍率而改變，不僅增加掃描時的穩定性且將掃描結構簡化。並使用20個老鼠切片來做初步的儀器驗證，得到靈敏度88.9%及特異度87.6%。此外，分別使用雞尾酒方法與組織分層法診斷29個及34個人類病理切片，來驗證這套系統的臨床適用性，前者得到靈敏度90%及特異度87.8%，後者得到靈敏度94.12%及特異度91.18%。最後，本研究也將此系統結合光纖內視鏡，做初步的臨床in-vivo實驗。

摘要(英)

The 5-years survival rate of oral cancer is approximately 57%; however, if the oral cancer is discovered in the early stage, the 5-years survival rates can be increased to 80%. Biopsy is a conventional approach for diagnosing oral cancer. In addition to allow pathologists to diagnose the stage of cancer accurately, a biopsy also allows them to prescribe the most appropriate treatment. However, the difference in experience and subjectivity between pathologists might affect their diagnostic accuracy. Therefore, the pathologist hopes to find an approach to quantitatively diagnose oral cancer.
A microscopic hyperspectral imaging system (MHIS), which can simultaneously present the tissue image and the spectral information of each pixel on an image, has been developed to aid physician to quantitatively diagnose oral cancer. However, the MHIS has some hardware issues, such as the unstable scanning mechanism, complex mechanical structure, and inconvenient alignment. Hence, this study develops a novel embedded relay lens microscopic hyperspectral imaging system (ERL-MHIS), which is composed of the proposed ERL-HIS and a microscope, to improve the hardware issues by optical transferring the scanning plane from objective plane to imaging plane.
In this study, the development of the ERL-MHIS is discussed and the capability of the system is firstly demonstrated by diagnosing early stage oral cancer of twenty mice biopsies. The sensitivity and specificity are 88.9% and 87.6%, respectively. Then, twenty-nine oral cancer (fourteen in early stage) patient’s biopsies are diagnosed using cocktail approach, which diagnoses the biopsy according to the patient’s condition. The sensitivity and specificity are 90% and 87.8%, respectively. Finally, the tissue stratification approach, which combines the morphological and spectral information of biopsy to discriminate cancer form normal biopsies, is used to diagnose thirty-four patient’s biopsies. The sensitivity and specificity are 94.12% and 91.18%, respectively. Moreover, we also connect the ERL-HIS with the fiberscope for in-vivo diagnosis.

關鍵字(中)

★ 超頻譜
★ 顯微鏡
★ 癌症
★ 切片
★ 活體

關鍵字(英)

★ Hyperspectral
★ Microscopy
★ Cancer
★ Biopsy
★ in-vivo

論文目次

中文摘要 I
Abstract II
Acknowledgements III
Contents IV
List of Figures VII
List of Tables XI
Chapter 1. Introduction 1
Chapter 2. Spectroscopy 4
2-1 Introduction of spectroscopy…………………………………………………………….15
2-2 Classification of spectroscopy by the interaction between radiation and substance ..……5
2-2-1 Absorption spectroscopy………………………………..…………………………5
2-2-2 Emission spectroscopy………………………………………………………..…...6
2-2-3 Scattering spectroscopy……..……………………………………………………..9
Chapter 3. Fundamentals of light-matter interactions ….12
3-1 Interactions between light and molecule………………………………………………...12
3-2 Interactions between light and cells……………………………………………………..13
3-3 Interactions between light and bulk matter……………………………………………...15
3-4 Interactions between light and tissues…………………………………………………...16
Chapter 4. Optical imaging modalities for biomedical applications 19
4-1 Optical imaging modalities classified to absorption spectroscopy……………………...19
4-1-1 Transmission microscopy………………………………………………………...19
4-2 Optical imaging modalities classified to scattering spectroscopy………………………21
4-2-1 Optical coherence tomography (OCT)…………………………………………...21
4-2-2 Confocal microscopy…………………………………………………………….23
4-2-3 Coherent anti-stokes Raman scattering (CARS)…………………………………24
4-3 Optical imaging modalities classified to emission spectroscopy……………………….25
4-3-1 Multiphoton microscopy…………………………………………………………25
4-3-2 Fluorescence lifetime imaging microscopy (FLIM)……………………………..27
4-3-3 Multispectral image………………………………………………………………29
4-3-4 Hyperspectral image……………………………………………………………...30
Chapter 5. The proposed novel hyperspectral imaging systems 36
5-1 Introduction of proposed in-vitro and in-vivo hyperspectral imaging systems………….36
5-2 Disadvantages of conventional microscopic hyperspectral system……………………..36
5-3 Finite conjugate relay lens………………………………………………………………39
5-3-1 Design principle………………………………………………………………….39
5-3-2 Simulations of relay lens…………………………………………………………40
5-3-3 Experiments of relay lens………………………………………………………...43
5-4 Embedded Relay Lens Microscopic Hyperspectral Imaging System (ERL-MHIS)……45
5-4-1 Operational principle of ERL-HIS……………………………………………….45
5-4-2 Operational Principle of ERL-MHIS…………………………………………….47
5-4-3 Analysis and calibration of ERL-MHIS………………………………………….51
5-4-4 Experiments and results of ERL-MHIS………………………………………….52
5-5 Embedded Relay Lens Fiberscopic Hyperspectral Imaging System (ERL-FHIS) ……...56
Chapter 6. ERL-MHIS for in-vitro oral cancer examination 59
6-1 Introduction of oral cancer………………………………………………………………60
6-2 Mice oral cancer biopsy examination…………………………………………………...61
6-2-1 Biopsy preparation and data collection…………………………………………..61
6-2-2 Data processing and analytical methods…………………………………………63
6-2-3 Results and discussion……………………………………………………………65
6-3 Human oral cancer biopsy examination by cocktail approach………………………….70
6-3-1 Patient biopsy preparation………………………………………………………..70
6-3-2 Five spectral methods for diagnosing oral cancer………………………………..73
6-3-3 Cocktail approach………………………………………………………………...75
6-3-4 Results and discussions…………………………………………………………..76
6-4 Human oral cancer biopsy examination by tissue classification………………………..81
6-4-1 Patient biopsy and image preparations…………………………………………...81
6-4-2 Cell classification on spectral domain……………………………………………82
6-4-3 Tissue stratification analysis on spatial domain………………………………….86
6-4-4 Results and discussions………………………………..………………………....92
Chapter 7. ERL-FHIS for in-vivo oral cancer examination 97
7-1 Patient samples preparations and image acquisition………………………………….....97
7-2 Data processing and analysis…………………………………………………………....97
7-3 Results and discussions………………………………………………………………..100
Chapter 8. Conclusions and future works 102
List of References 106
Personal publications 116

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指導教授

歐陽盟、李正中(Mang Ou-Yang Cheng-Chung Lee)

審核日期

2014-11-10

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