修飾負電微米粒子之紙晶片應用於豬X-Y精子分選

隨著畜牧業的高度發展和乳製品的高度需求，牲畜經濟儼然變成一個炙手可熱的方向，但是現今的研究方向和已存在的方式卻不十分著墨，因此本篇論文想要發展一個方式用來分選豬的XY精子，讓牲畜的下一代可以決定是生公的或母的，如此一來便有非常龐大的經濟價值。生醫晶片已經是非常成熟的技術，舉凡蛋白質、細菌和病毒，有無數的文獻利用晶片來達成不管是篩選或是檢測的目的，因此本篇研究想要利用晶片來達成分選精子的目標，但同時又要開創一個利用新的基材-紙來製作。傳統用來分選的方法如pure sperm和白蛋白液注所得出的分選效率都不十分顯著，但像是效果很好的電泳分離法和流式細胞儀，儘管效果明顯，由於實驗過程中會傷害精子，導致回收出來的精子存活率低，因此要如何在提高分選效率的同時又能維持精子的存活率也是本篇研究的重點。利用製成簡易的優勢來製作紙晶片，過程中只需使用印表機和烘箱在紙上製造出流道，再利用表面修飾帶負電的微米例子，最後再使用靜電產生器對表面做改質，就能完成整個晶片。檢測時只需把原液直接滴入晶片，靜待一分鐘就能完成實驗，最後再把跑完晶片的液體拿去流式細胞儀做分析。本研究能夠達到將死活精子分離開來，而且能夠確保活精子裡面都是品質比較好的，最後X精子和Y精子的分選比例結果可以達到61:39。

關鍵字

紙晶片；精子； XY分選

並列摘要

As long as the blooming of livestock and high demand for dairy, livestock economy seems to turn into a hot direction. But the research and methods nowadays are not very value, so this paper wants to develop a method to differentiate the X and Y sperm of farrow. In that we can choose the offspring of livestock, which will become a huge property. Biochip is already become a mature way to select or detect not only protein but also germ and virus. So this paper wants to use the theory of biochip to differentiate the sperm in X and Y, but also wants to open up a new base-paper to make the chip. Traditional methods for differentiation efficiency like puresperm and albumin are not very powerful. Although the high efficiency ways like electrophoresis and flow cytometric will harm to the sperm. Because electrophoresis way will use electricity direct among the sperm so it will harm to them and flow cytometerichave to use the fluorescent dye. So these ways will make the survival rate of sperm be very low. In conclusion, this paper will focus on how to raise not only the differentiation efficiency but also the survival rate. This paper has the advantage of easy fabrication, just need printer and oven to make the chip on the filter paper. After the fabrication, spin the polystyrenebeads that modified with negative functional groups on the surface of the chip. At last, use plasma treatment on the chip. When run the experiment, just drop the sperm of farrow on the chip, and then wait for one minute to collect the liquid in the specific position. At last, use the flow cytometric to analysis. This research can achieve to separate the live sperm from dead and live sperm is in good quality. The differentiation efficiency can reach to 61:39 between X and Y sperm.

並列關鍵字

無資料

參考文獻

1. 胡一君、游智勝、林明瑜、胡恆蒼、陳順源，“微型生醫晶片發展與應用”，儀科中心簡訊 69 期，台灣，中華民國94年。

2. T. Ichiki et al, “Development of bio-MEMS devices for single cell expression analysis,” in Microprocesses and Nanotechnology Conference, 2001 International, 2001, pp. 190-191.

7. Satoru Kaneko et al, “Human X- and Y-bearing sperm differ in cell surface sialic acid content”, Biochemical and biophysical research communications, 1984

8. Dr Hamid Gourabi et al, “Separation of X- and Y-bearing human spermatozoa by sperm isolation medium gradients evaluated by FISH”, Reproductive BioMedicine, 2009

10. D.L. Garner et al, “Hoechst 33342: The dye that enabled differentiation of living X-and Y-chromosome bearing mammalian sperm”, Theriogenology, 2009

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修飾負電微米粒子之紙晶片應用於豬X-Y精子分選

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