現今，世界各地的醫療機構都會使用富含血小板的血漿來治療患有某些先天性和後天性血小板功能不全的患者。但血小板只能在室溫（25℃）下儲存血小板，這是因為低溫環境會使血小板無法長時間在體內循環。因此，血小板難以用於“先天性血小板功能缺陷”的治療。然而，當血小板在室溫下儲存時，血小板可能容易受到細菌污染，從而縮短血小板的保質期，造成血小板的浪費。在本研究中，我們懷疑血小板不能在低溫下儲存的部分原因可能是由於血小板衍生的細胞外囊泡（EV）。
我們實驗室開發了一種免疫親和紙基裝置，可以用於分離的細胞外囊泡。因此，在本研究中，我們使用紙基裝置連續捕獲源自血小板的細胞外囊泡並必較不同儲存溫度的結果，以闡明血小板、血小板細胞外囊泡和溫度之間的關係。而最終目標將是解決血小板儲存問題。
在分析實驗初步結果後，我們發現紙基裝置本身會影響血小板的行為。因此，此論文另開發一種由PDMS製作的免疫親和多孔膜裝置，以取代紙基裝置。我們將其命名為血小板細胞外囊泡捕手（PEVC）。

Nowadays, medical institutions around the world will use platelet rich plasma to treat patients who have had some nature and nurture symptoms that have led to platelet insufficiency. But it can only store platelet at room temperature (25℃) that was because the low-temperature environment will prevent platelets from circulating in the body for a long time. Thus, the platelet was difficult to be used for the treatment of “congenital platelet function defects”. However, when the platelet was stored at room temperature, the platelet may be prone to bacteria contamination, which shortens the shelf life of the platelets and results in the waste of platelets. In this report, we suspected that the reason why platelets cannot be stored at low temperature may be partially due to extracellular vesicles (EV) derived from the platelet.
In the beginning, we have developed an immunoaffinity paper-based device that can isolate variable extracellular vesicles. So, in this study, we have used the paper-based device to continuously catch extracellular vesicles derived from the platelets and stored under different conditions to elucidate the relationship between platelet, platelet EV, and temperature. Then a final goal would be to solve the platelet storage problem.
After analysing the experiment results, we have discovered that the paper-based device itself would influence the platelet behaviour. Thus, a new design of immunoaffinity porous film would make by PDMS to replace the paper-based device. And we named it the platelet extracellular vesicle catcher (PEVC).

Abstract..i
List of figures..iv
List of tables..vi
Purpose..1
Chapter..1
Introduction..2
1-1 Extracellular vesicle..2
1-2 Platelet extracellular vesicle..5
1-3 Platelet characteristic..8
1-4 Cold-stored platelets physiology..10
1-5 Extracellular vesicle isolation process..11
1-6 Technology for Polydimethylsiloxane (PDMS) microfluidic..13
1-7 Self-assembled monolayers technology..14
Chapter 2. Experiment design..17
2-1. Paper-based device fabrication and function test..18
2-2. Optimal the electron microscope experiment..20
2-3. Optimal PEV isolation process..20
2-4. EV absorption ability test for experiment instrument..20
2-5. Platelet aggregation test by spectrophotometer..21
2-6. Main experiment..21
2-7. PEVC fabrication and function test..23
Chapter 3. Experiment method..25
3-1 HEK293T PalmGP EV isolation..25
3-2 Paper-based device fabrication and function test..26
3-3 Scan electron microscope image for EV on paper-based device..32
3-4 Optimal PEV isolation process..34
3-5 EV absorption ability test for experiment instrument..36
3-6 Platelet aggregation test by spectrophotometer..38
3-7 Main experiment..39
3-8 PEVC fabrication, surface modification and function test..41
Chapter 4. Results and discussion..46
4-1. Function test of paper-based device by PE-biotin..46
4-2. Saturation test of paper-based device by PE-biotin..47
4-3. Paper-based device EV catching test..48
4-4. Scan electron microscope image for EV..49
4-5. Optimal PEV isolation process..51
4-6. EV absorption ability on 6-well plate..51
4-7. EV pass and absorption ability on the 1μm insert..52
4-8. Main experiment..53
4-9. PEVC fabrication and function test..58
Chapter 5. Summary..62
Chapter 6. Future work..64
Chapter 7. Reference..66

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