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離心微流體平台應用於全血中自然殺手細胞分離與數位計數 = = Isola...

洪宗源

離心微流體平台應用於全血中自然殺手細胞分離與數位計數 = = Isolation and Digital Counting of Natural Killer Cells from Blood Via Centrifugal Microfluidic Platform /

紀錄類型:	書目-語言資料,印刷品 : Monograph/item
正題名/作者:	離心微流體平台應用於全血中自然殺手細胞分離與數位計數 =/ 洪宗源.
其他題名:	Isolation and Digital Counting of Natural Killer Cells from Blood Via Centrifugal Microfluidic Platform /
其他題名:	Isolation and Digital Counting of Natural Killer Cells from Blood Via Centrifugal Microfluidic Platform.
作者:	洪宗源
出版者:	雲林縣 :國立虎尾科技大學 , : 民113.07.,
面頁冊數:	[12], 68面 :圖, 表 ; : 30公分.;
附註:	指導教授: 郭如男.
標題:	COMSOL Multiphysics. -
電子資源:	電子資源

離心微流體平台應用於全血中自然殺手細胞分離與數位計數 = = Isolation and Digital Counting of Natural Killer Cells from Blood Via Centrifugal Microfluidic Platform /
洪宗源

離心微流體平台應用於全血中自然殺手細胞分離與數位計數 =Isolation and Digital Counting of Natural Killer Cells from Blood Via Centrifugal Microfluidic Platform /Isolation and Digital Counting of Natural Killer Cells from Blood Via Centrifugal Microfluidic Platform.洪宗源. - 初版. - 雲林縣 :國立虎尾科技大學 ,民113.07. - [12], 68面 :圖, 表 ;30公分.

指導教授: 郭如男.

碩士論文--國立虎尾科技大學自動化工程系碩士班.

含參考書目.

自然殺手細胞對免疫系統至關重要，但傳統密度梯度分離方法效率低且花費時間長。而離心微流體技術具有少量樣本、微小體積、低成本和快速分離的優勢，能精確的分離目標細胞，實現高效率細胞分離和檢測，為免疫細胞的研究提供了一個極為實用的工具。本研究結合方波型混合流道和細胞分選流道於離心平台上進行全血中自然殺手細胞的分離。在微流體晶片上首先利用方波型混合流道混合血液樣本和細胞團聚試劑，混合完成後會在靜置槽中等待樣本和試劑充分的結合，讓血液中除了自然殺手細胞外的所有細胞團聚成尺寸較大的玫瑰花結。接著利用細胞分選流道將自然殺手細胞分離出來到收集槽中，分選完成後利用Python OpenCV建構的視覺辨識系統來應用於數位計數自然殺手細胞的數量，將目標收集槽的螢光影像進行多項影像處理，包括灰階轉換、二值化、數量偵測等步驟，檢測結果表明能準確計算目標粒子的數量得到系統的決定係數R2 = 0.9979。微流體裝置首先透過COMSOL Multiphysics模擬來驗證可行性，模擬結果顯示方波型混合流道轉速在1500 rpm時有最佳混合效率為93.5%，細胞分選流道轉速在3000 rpm時有最佳分離效率為90.0%。晶片透過黃光微影製程技術來製作離心微流體裝置的SU-8母模，再透過PDMS翻模和接合來完成離心微流體裝置，將製作完成的離心微流體裝置應用於方波型流道混合和全血中自然殺手細胞的分離實驗，實驗結果顯示方波型混合流道的混合效率為93.3%，全血中自然殺手細胞的分離效率為84.9%、純度為80.0%及存活率為93.9%。.

(平裝)Subjects--Topical Terms:

1450033
COMSOL Multiphysics.

離心微流體平台應用於全血中自然殺手細胞分離與數位計數 = = Isolation and Digital Counting of Natural Killer Cells from Blood Via Centrifugal Microfluidic Platform /
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245 1 0 $a 離心微流體平台應用於全血中自然殺手細胞分離與數位計數 = $b Isolation and Digital Counting of Natural Killer Cells from Blood Via Centrifugal Microfluidic Platform / $c 洪宗源.
246 1 1 $a Isolation and Digital Counting of Natural Killer Cells from Blood Via Centrifugal Microfluidic Platform.
250 $a 初版.
260 # $a 雲林縣 : $b 國立虎尾科技大學 , $c 民113.07.
300 $a [12], 68面 : $b 圖, 表 ; $c 30公分.
500 $a 指導教授: 郭如男.
500 $a 學年度: 112.
502 $a 碩士論文--國立虎尾科技大學自動化工程系碩士班.
504 $a 含參考書目.
520 3 $a 自然殺手細胞對免疫系統至關重要，但傳統密度梯度分離方法效率低且花費時間長。而離心微流體技術具有少量樣本、微小體積、低成本和快速分離的優勢，能精確的分離目標細胞，實現高效率細胞分離和檢測，為免疫細胞的研究提供了一個極為實用的工具。本研究結合方波型混合流道和細胞分選流道於離心平台上進行全血中自然殺手細胞的分離。在微流體晶片上首先利用方波型混合流道混合血液樣本和細胞團聚試劑，混合完成後會在靜置槽中等待樣本和試劑充分的結合，讓血液中除了自然殺手細胞外的所有細胞團聚成尺寸較大的玫瑰花結。接著利用細胞分選流道將自然殺手細胞分離出來到收集槽中，分選完成後利用Python OpenCV建構的視覺辨識系統來應用於數位計數自然殺手細胞的數量，將目標收集槽的螢光影像進行多項影像處理，包括灰階轉換、二值化、數量偵測等步驟，檢測結果表明能準確計算目標粒子的數量得到系統的決定係數R2 = 0.9979。微流體裝置首先透過COMSOL Multiphysics模擬來驗證可行性，模擬結果顯示方波型混合流道轉速在1500 rpm時有最佳混合效率為93.5%，細胞分選流道轉速在3000 rpm時有最佳分離效率為90.0%。晶片透過黃光微影製程技術來製作離心微流體裝置的SU-8母模，再透過PDMS翻模和接合來完成離心微流體裝置，將製作完成的離心微流體裝置應用於方波型流道混合和全血中自然殺手細胞的分離實驗，實驗結果顯示方波型混合流道的混合效率為93.3%，全血中自然殺手細胞的分離效率為84.9%、純度為80.0%及存活率為93.9%。.
520 3 $a Natural killer cells are critical to the immune system, but the traditional density gradient separation method is inefficient and time-consuming. With the advantages of small sample size, small volume, low cost and rapid separation, the centrifugal microfluidic technology can accurately separate target cells and achieve high-efficiency cell separation and detection, providing an extremely practical tool for the study of immune cells. In this study, natural killer cells in blood were separated by combining the square wave mixing channel and cell sorting channel on the centrifugal platform. Firstly, the blood sample and a cell aggregation reagent were mixed in the a mixing channel, and then the sample and the reagent were fully combined in a standing tank, so that all cells in blood except natural killer cells can be aggregated into a larger rosette. Then, the natural killer cells were separated in the cell sorting channel and then stored into the collection tank. After sorting, the visual identification system constructed by Python OpenCV was used to count the number of natural killer cells digitally. The fluorescence image of the target collection tank was processed by several steps, including gray-scale conversion, binarization, quantity detection and so on. The detection results showed that the system’s determination coefficient R2 of 0.9979 can be obtained by accurately calculating the number of target particles. Firstly, the feasibility of the microfluidic device was verified by COMSOL Multiphysics simulation. The simulation results showed that the mixing efficiency was the best (93.5%) when the velocity of the mixing channel was 1500 rpm, and the separation efficiency was the best (90.0%) when the velocity of the cell sorting channel was 3000 rpm. The SU-8 master mold of the centrifugal microfluidic device was made by the chip through the yellow-light photolithography process technology, and then the centrifugal microfluidic device was completed through PDMS mold turnover and bonding. The device was applied to the test. The test results showed that the mixing efficiency of the mixing channel was 93.3%, the separation efficiency, the purity and the survival rate of natural killer cells in blood were 84.9%, 80.0% and 93.9%, respectively..
563 $a (平裝)
650 # 4 $a COMSOL Multiphysics. $3 1450033
650 # 4 $a natural killer cell. $3 1450047
650 # 4 $a mixing channel. $3 1450046
650 # 4 $a centrifugal microfluidic. $3 1450045
650 # 4 $a 自然殺手細胞. $3 1450028
650 # 4 $a 混合流道. $3 1450044
650 # 4 $a 離心微流體. $3 1450043
856 7 # $u https://handle.ncl.edu.tw/11296/v9xav2 $z 電子資源 $2 http