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標題: 血液於旋轉微流道晶片沉降分離與型態之可視化實驗
Human peripheral blood sediments、separates and forms in centrifugal microchip : visualization experiments
作者: 梁博欣
Laing, Po-Shin
關鍵字: Microfluidics channel;旋轉微流道;visualization;Hct(Hematocrit);cells form;可視化;血球容積計;血球型態
出版社: 機械工程學系所
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本實驗設計之旋轉微流道生物晶片,可以一次檢測4組檢體,並利用三種實驗機制來達成檢測目的,省去換血片之時間與步驟。本研究以旋轉時產生的離心力驅動血液,探討血液在旋轉微流道晶片上的分離率與血球現象。實驗採用CNC加工PMMA(Polymethyl methacrylate)方式製作實驗晶片,再經由熱壓的方式將晶片結合。流道的設計包含Metering chamber與Observation chamber結構,流道深度分別為100、150與300 μm,寬度則為100至300 μm的範圍。實驗主要觀查晶片上血液在Metering chamber中分離成血漿血球的比率與Observation chamber中血球之型態,同時藉由Hct(Hematocrit)實驗、光學偵測與螢光偵測的方式觀察。Hct實驗則以He-Ne Laser及光二極體同步觸發CCD攝影機擷取影像;光學偵測則以光學顯微鏡觀察影像;螢光偵測則以螢光顯微鏡並搭配旋轉平台與Cooling CCD攝影機擷取影像。本實驗發現,在Hct實驗下晶片的旋轉方向為順時針旋轉,轉速為3000 rpm旋轉時間為3分鐘,此時全血分離成血漿血球的比率可以當作血球容積計來檢測貧血。在螢光偵測時,則是利用濃度為1 mM的螢光染劑在螢光顯微鏡放大150倍下來判定血球是否為活體細胞。在光學偵測時晶片上片厚度必須在0.5 mm才可以觀測到放大400倍的效果,實驗中必需利用DPBS(Dulbecco''s Phosphate buffered Saline without Calcium or Magnesium)溶液將血液稀釋成30倍才可清楚看見血球的形態,利用血球型態則可以檢測肝病與腎病。

This study reports three experiments to detect four specimens all at once in the rotating microfluidic chip. The advantage of the experiments is saving the time and steps of changing blood smears. This experimental study utilizes the centrifugal force for driving the bloodstream to study the human whole blood separation and cells phenomenon in the chip. The testing chip is fabricated on PMMA (Polymethyl methacrylate) using CNC machining and hot plat bonding. The microfluidics of the testing chip structure composed of metering chamber and observation chamber. The channel depths of the chambers were 100, 150 and 300 μm and the wide ranges were 100 to 300 μm. The Hct (Hematocrit) experiment, using optical microscope detection and fluorescence microscope detection, could be used to complete the experiment at the same time. The Hct experiment utilizes He-Ne laser and photodiode to trigger the CCD camera to take the image. When the chip rotated at rotation speed of 3000 rpm clockwise for 3 minutes, the results show the microfluidics can be used as hematocrit to detect anemia. In the detection with fluorescence microscope at 150 X magnifies live cells of blood can be distinguished. At a higher magnification of 400 X, forms of cells can be clearly observed. Under above-mentioned conditions, we could detect the liver complaint, nephropathy and anemia by distinguishing forms of cells in the rotating microfluidic chip.
其他識別: U0005-2608200816534000
Appears in Collections:機械工程學系所

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