微流體生醫晶片用來分選精子的方法粗略可分為利用穩定層流和利用逆向流場兩類：前者利用穩定層流的方式將具有活動力的精子從起始層流分出在後端出口收集，但缺點是收集到具有活動力的精子不能區分其活動力的高低；後者利用逆向流場的方式由於精子有能夠抵抗流場的特性，因此不具有活動力的精子幾乎可以完全帶走，但缺點是逆向流場所產生的剪應力會削弱精子的活動能力。
本篇所提出之漸擴型微流道晶片，利用精子會抵抗流場之特性採順向流場將精子從進樣口處帶往分選區，縮短精子在流道內運動之距離。流體進入流道內後會因為流道寬度不同而導致流體速度減緩，流道寬度從100μm漸擴至1000μm，漸擴角度為5∘，具有活動力的精子有抵抗流場之特性能停留在流道內，而不具有活動力的精子則會隨著流體被帶往出口處。實驗方法依液體驅動力可分為液壓差被動式及幫浦穩壓主動式兩種方式。液壓差被動式則是以液壓差為2mm高度之時分選區最窄處可產生流體速度為240μm/s，依據精子的運動速度將分別停流於分選區不同之位置；幫浦穩壓主動式則是設定注射式幫浦之流量在0.0648ml/hr，可依公式計算出分選區最窄處產生流速為180μm/s。分選後依取樣方式可分為分層推出和側枝取樣兩種方式，分層推出法為在流道內分別給予一股流體，第一次先將不具活動力的精子推出，第二次再將具有活動力的精子推出，側枝取樣法則是直接從分選區旁加上兩側枝，當精子分選完成後直接從旁取出。實驗結果發現，以被動式載入精子分選率可達78.7%，而取樣方法則為側枝取樣分選率可達84.8%。
本研究所提出之微流體系統晶片可避免削弱精子的活動力，並可控制取出的精子活動力品級，為多重門檻篩選之微流體系統晶片。

The methods applied in separating sperms in microfluidic bio-chip can divide in steady laminar flow and reversing flow. Due to motile sperms can across the original laminar to the selection laminar, the motile sperms can be collected in the outlet. The disadvantage of the former way is that we can not tell the sperms motility apart. Sperms can against the flow by nature property, thus, separating the motile sperms by using reversing flow can select motile sperms. However the shearing force in the reversing flow will cripple the sperms motility. The thesis presents a novel chip that diffuser type channel will slow down the fluidic velocity in channel, the motile sperm will stay in the chamber owe to their nature property and the non-motile ones will drift to outlet with flow. By controlling the flow velocity in channel can select the motility of sperm which stayed in channel. The motility of sperm will not cripple, the classification of sperm can be determined as well.

The thesis presents a novel diffuser type microfluidic chip, based on the sperm can against the fluidic. We use the forward flow instead of the reverse flow to reduce the distance which sperm swims to expanding area. The fluidic velocity in channel will decrease with the vary channel width. The channel width in 100μm increase to 1000μm and the diffuse angle is 5∘. The motile sperm can against flow and stay in the channel while non-motile sperm will drift to the outlet. The fluidic driving force can be concluded into “pressure height passive way” and “continuous steady pressure active way”. The pressure height in 2mm can generate the fluidic velocity in 240μm/s in the passive way and the sperms will stay at the specific position based on their positions. Setting the syringe pump flow rate at 0.0648ml/hr in the active way and the velocity can be calculated into 180μm/s. There are two way can be applied to take the sperm out. One way is by controlling the fluid velocity. The nonmotile sperm can be washed out at the first time while the motile sperm can be washed out at the second time. Another way is adding two branches at the expanding area and the sperm will be taken out from the branches. As the results show, the live and dead ratio in passive way reaches in 78.7% and the live sperm taken from the branches ups to 84.8%.

The thesis presents a microfluidic device can separate the motile sperm but not to hurt the sperm moltility, moreover the sperm motility can be classfied. It can be considered as multi-criterion separation chip.

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