外吐體純化與檢視系統研製

The Development of an Exosomes Purification and Inspection System

古原煌(Yuan-Huang Gu)

指導教授 ： 章明

中原大學/工學院/機械工程研究所/碩士(2017年)

https://doi.org/10.6840/cycu201700661

摘要

癌症多年來為國人十大死因之榜首，若能提早發現並予以治療，將使存活機率大大提升。由於當癌細胞開始增生或將要進行移轉時，存在於體液中的外吐體會異常大量分泌，若能開發出一套簡易式檢測體液外吐體數量變化的設備，就可能達到癌症預警之效。
本研究研製了一套結合外吐體純化稀釋取樣與奈米粒子數量檢視之系統，先透過包覆聚乙二醇之四氧化三鐵奈米粒子抓取血清內部之雜蛋白質，利用磁力將粒子吸附至底部以取得純化之外吐體上清液，再將上清液與去離子水稀釋混合，最後將稀釋後待測液放入奈米粒子數量檢視系統中，即可藉由雷射光照射外吐體粒子產生之瑞利散射光檢測外吐體的數量。
本研究中的檢視系統係由傳統的顯微鏡改裝而成，使用波長405nm雷射作為光源，並以3D列印技術製作暗場觀測平台。實驗結果顯示在各種濃度中所測得的外吐體數量與商用Nanosight所測得的數量值誤差在10%內，且此外吐體純化稀釋取樣設備能夠在3分鐘內自動完成純化稀釋取樣與數量檢視，故此低成本系統有機會開發成為一套幫助人類早期發現癌細胞的儀器設備。

關鍵字

外吐體 ； 癌症預警 ； 瑞利散射 ； 低成本

並列摘要

For a long time, cancer is a disease that leads to paramount mortality, if we earlier can detect and give appropriate treatment, the more higher survival rate will be greatly improved. When cancer cells begin to hyperplasia or transfer, exosomes will be abnormally secreted in the body. So, if we can develop a simple inspection to detect the quantity of exosomes, perhaps that we just build an early warning for cancers.
In this study, we developed a system for purification, dilution sampling of exosomes, and inspection of the number of nanoparticles. First of all, capturing the proteins inside the serum through Polyethylene glycol (PEG)-coated Fe_3 O_4 nanoparticles, then the particles were attached to the bottom by using magnetic force to obtain the exosomes supernatant, the supernatant was diluted with deionized water, and finally put the diluted measuring fluid into the nanoparticles inspection system, the exosomes was measured by rapid image processing after Rayleigh scattered light produced by laser irradiation of exosomes.
In this study, the Exosome Purification and Dilution Sampling System are not only able to automatically complete the purification and dilution sampling within 3 minutes, but also can effectively reduce the human factors. Besides, the inspection system only need to improve the traditional microscope, and use laser and 3D printing technology to create a dark field observation platform, to inspect nanoparticles quickly. It has low-cost and easy to popularize characteristics, have the opportunity to develop into an equipment to help humans early detection of cancer cells.

並列關鍵字

Exosome ； cancer warning ； Rayleigh scattering ； low cost

參考文獻

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