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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/79695完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 朱士維(Shi-Wei Chu) | |
| dc.contributor.author | Hung-Yu Chen | en |
| dc.contributor.author | 陳鴻羽 | zh_TW |
| dc.date.accessioned | 2022-11-23T09:07:51Z | - |
| dc.date.available | 2022-02-21 | |
| dc.date.available | 2022-11-23T09:07:51Z | - |
| dc.date.copyright | 2022-02-21 | |
| dc.date.issued | 2022 | |
| dc.date.submitted | 2022-02-07 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/79695 | - |
| dc.description.abstract | 為了了解大腦是如何運作的，全腦功能性成像技術是不可或缺的。在各種技術中，光學顯微術由於能捕抓個別神經元訊號的微米級空間解析度與毫秒級時間解析度而十分受歡迎，但是它典型的穿透深度僅有一毫米。因此，對於全腦研究，果蠅成為最具潛力的模式生物，它的腦小到理論上可以被光學顯微鏡所穿透。此外，它將近一半的腦神經結構性圖譜已經在Flycircuit被繪製，這個高比例完成度的結構圖譜成為果蠅功能成像珍貴的參考。然而，實際上，我們發現光學顯微鏡在果蠅腦中遇到意外的光損害，這造成我們無法對其做全腦成像，而其原因就源自於果蠅腦中的氣管。氣管裡的空氣與周圍組織的折射率的巨大差異，使得光遭遇強烈的像差/散射，因此無法抵達大腦的深層。 在本篇研究中，我們從醫學研究中的液體通氣(liquid ventilation)取得靈感，提出三種液體填充的方法試圖能降低氣管所造成的像差/散射。在所有方法中，滲透方法(osmosis method)成效最好，藉由間接改變果蠅腦內的滲透壓，使組織液因滲透壓差而流進微氣管裡取代原本的空氣。我們經由共厄焦顯微鏡及電刺激來分別檢視滲透方法對於影像穿透深度的提升及應用在功能性成像上的可行性。不過，我們的結果顯示，並非所有果蠅腦成像的穿透深度都有增強，我們歸因於那些會影響微氣管中液體的複雜生理現象。這這些現象需要活體氣管成像的技術，可能可以進一步檢測滲透方法。總而言之，這個研究為我們提供了能增加果蠅腦內光學穿透深度的方法，並且為全腦功能連接體研究鋪路。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-23T09:07:51Z (GMT). No. of bitstreams: 1 U0001-0602202218544900.pdf: 12072777 bytes, checksum: 1c80532b5a6a34717d167fed007007c4 (MD5) Previous issue date: 2022 | en |
| dc.description.tableofcontents | "口試委員會審定書 i 謝辭 ii 摘要 iii Abstract iv Contents vi Figure list ix Table list xiii Chapter 1. Introduction 1 1.1. From brain to connectome 1 1.2. Contemporary methods for whole brain functional connectome study 4 1.2.1. Technique for functional connectome: Why optical microscopy? 4 1.2.2. Animal models for whole brain connectome study: Why choosing Drosophila? 7 1.3. Bottleneck and approaches for Drosophila whole brain functional connectomics 9 1.3.1. Bottleneck: Shallow penetration depth due to aberration/scattering from trachea 11 1.3.2. Published approaches for better penetration in Drosophila brain 15 1.3.3. New approach for reducing reduce aberration/scattering from trachea: liquid filling method 17 1.4. Aim: Enhance optical penetration depth in Drosophila brain - minimizing aberration/scattering from trachea by liquid filling method 19 Chapter 2. Principle 20 2.1. Limited penetration depth in Drosophila brain: Effect from aberration and scattering 20 2.1.1. Effect from aberration and scattering in Drosophila brain 20 2.1.2. Quantify the penetration depth 23 2.2. Enhance penetration via aberration/scattering elimination: Osmosis method 26 2.3. Optical imaging technique for checking the penetration depth: Confocal laser scanning microscope 33 2.4. Optical imaging technique for imaging tracheas: Third harmonic generation microscopy 36 Chapter 3. Method 39 3.1. Sample preparation 39 3.1.1. Drosophila 39 3.1.2. Mounting the fly 40 3.2. Optics setup 43 3.2.1. Confocal laser scanning microscope 43 3.2.2. Third-harmonic generation microscope 43 3.3. Improve the penetration depth: liquid filling method 45 3.3.1. Liquid selection: PFCs, PFC emulsion, PBS 45 3.3.2. Filling method: Capillary method, Injection method, Osmosis method 48 3.4. Extend to functional imaging: survival test 53 3.4.1. Stimulation setup for survival test 53 3.4.2. Stimulation protocol 57 3.5. Experiment protocol for osmosis method and survival test 58 3.6. Analysis method for penetration depth 59 Chapter 4. Experiments and Results 65 4.1. THG imaging for trachea 65 4.2. Liquid filling method: PFCs and capillary method 67 4.3. Liquid filling method: PFC emulsion and injection method 69 4.4. Liquid filling method: PBS and osmosis method 71 4.4.1. Drosophila brain imaging with 100% PBS 71 4.4.2. Brain imaging in different concentration PBS from 90% to 70% 73 Chapter 5. Discussion 78 5.1. The feasibility and the advantage of THG microscopy for imaging trachea 78 5.2. The reliability and the shortcomings of our liquid-filling model 81 5.2.1. Compare our model with the published result 81 5.2.2. Compare our model with our results 84 5.3. Penetration enhancement via osmosis method 86 Chapter 6. Conclusion and Perspective 90 Chapter 7. Supplementary 91 7.1. Detailed results of brain images with hypotonic mounting medium 91 7.2. Optimal wavelength for Drosophila brain imaging 107 Reference 111 " | |
| dc.language.iso | en | |
| dc.title | 藉由液體填充法減少氣管像差/散射以增加果蠅腦光學穿透深度 | zh_TW |
| dc.title | Enhance optical penetration depth in Drosophila brain – minimizing aberration/scattering from trachea by liquid filling method | en |
| dc.date.schoolyear | 110-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 江安世(Jeng-Wei Tjiu),朱麗安(I-Ping Tu),賈世璿(Snow H. Tseng) | |
| dc.subject.keyword | 功能成像,光學顯微術,果蠅,氣管,像差/散射,滲透壓, | zh_TW |
| dc.subject.keyword | Functional imaging,optical microscope,Drosophila,trachea,aberration/scattering,osmolarity, | en |
| dc.relation.page | 117 | |
| dc.identifier.doi | 10.6342/NTU202200302 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2022-02-09 | |
| dc.contributor.author-college | 理學院 | zh_TW |
| dc.contributor.author-dept | 物理學研究所 | zh_TW |
| 顯示於系所單位： | 物理學系 | |
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