近年來世界各地的新興感染症層出不窮，由於微生物不斷的突變，造成抗生素使用的極大困難。如何提升診斷效率與正確地使用藥物，是專家學者致力研究的領域。介電泳應用於操控細胞、微生物、病毒與DNA等的技術已有多例被結合在微流體晶片或感測器中，能夠快速與精準地操控並分析特定目標是技術伸張的要點。本論文將介電泳技術結合微流體晶片，進行微生物與其藥敏性的快速檢測。研究架構主要分為三個子題：(1)以灰階影像處理技術進行微生物的定量於介電泳晶片。生物微粒或細胞經由連續流體傳輸且被介電泳力影響而捕捉濃縮在特定區域，在最佳條件的控制下，能夠使捕捉效率達90%以上。實驗結果再以灰階影像處理技術進行量化，經換算之後便能得知樣本濃度。首先以聚苯乙烯微粒為樣本進行實驗，確認實驗的可行性之後；便以念珠菌種(Candida glabrata & Candida ablicans)進行生物檢體的量化實驗。樣本濃度的最佳檢測範圍約在105~107 CFU/mL，且實驗耗時不超過30分鐘。此實驗結果也與標準計數法的結果有高度的關聯性；因此，此系統對於微生物檢體的量化分析具有一定的可信度。(2)以介電泳法進行微生物的藥物敏感性試驗。此研究將介電泳技術應用於致病菌的抗藥性檢測與最小抑制菌濃度的判定，並希望達到快速且正確地診斷。以乙內醯胺類(-lactam)抗生素抑制細菌的細胞壁合成，且造成菌體延長的現象，造成細菌的介電泳特性產生變化。實驗結果發現當藥物濃度能夠抑制細菌生長時，其介電泳的特徵頻率會有顯著地變化；若是抗藥性菌株受藥物作用仍會持續進行細胞分裂，且菌體延長不明顯，故其特徵頻率並無改變。此方法不僅能夠辨知細菌的抗藥性與否，且在不同藥物濃度作用之後，能利用其特徵頻率的變化，快速判讀抗生素對於致病菌的最小抑制濃度。此外，整個實驗流程的耗時不超過兩個鐘頭，能夠大幅地縮短檢測時間。在檢測方法的可行性確定之後，進一步針對重要臨床致病菌的純化菌株(5株大腸桿菌、5株克雷白氏肺炎桿菌)進行藥敏性試驗，並與培養液稀釋法具有一致的結果。此方法已證明能適用於乙內醯胺類對革蘭氏陰性桿菌的藥敏性試驗且大幅縮短檢測的時間。此外晶片的製作十分簡易且所需成本低。針對革蘭氏陽性菌的檢測，未來也著手進行試驗。革蘭氏陰性菌(大腸桿菌與克雷白氏菌)為臨床感染症中的重要致病菌常帶有多重抗藥性，而乙內醯胺類抗生素為使用藥物的大宗。因此若能成功地應用在臨床診斷，相較於傳統的培養法可大幅縮短檢測時間(1-2小時)，對於醫師的用藥指引有很大的幫助，亦可減少藥物濫用情形。(3)微粒在波形結構上的電動力學之探討。此章節主要在討論微粒在波浪型結構上，受到非均勻的交流電場作用下，誘發電動力學的現象(介電泳與交流電滲流)。首先在指叉式電極陣列上施加交流電場，使電極上的感光薄膜材料受介電泳力作用而產生波形結構，且藉由施加的電壓大小來控制其振幅，最後以紫外光照射使薄膜固化。第二階段的實驗是將此波形結構應用於粒子的操控。藉由調整交流電場的頻率或波形，使得粒子受到介電泳力或交流電滲流的作用，進而達到粒子的傳輸、定位或濃縮等目的。另外，不同尺寸的粒子於波形結構上能受交流電滲流與拖曳力的作用而進行分離。未來希望能夠藉由設計不同的波形結構，進而應用在生物細胞的排列、定位、分離及濃縮。

Nowadays, emerging infectious diseases seem to appear in an endless stream all over the world, and the mutation of microbes makes problems in the use of antibiotics. How to improve the diagnostic efficiency and reduce the antibiotic misuse were important issues that researchers are studying. Dielectrophoresis (DEP) for the manipulation of cells, microbes, viruses and DNA, has been widely integrated into microfluidic chips or min devices for manipulating specific targets accurately. In this thesis, three subjects will be introduced, I. Microbial quantification based on grayscale image processing in a DEP chip The first part is the quantification of bioparticles based on the grayscale image processing (GIP) in the DEP-based microfluidic chip. The polystyrene beads were used as the sample particles to test for the feasibility of this approach; and then, the Candida cells were tested by GIP method to compare the standard counting of hemocytometer. The detection range of sample concentration is around 105-107 cells/ml, and the experimental time does not exceed 30 min. Ⅱ. Dielectrophoresis-based antibiotic susceptibility test The second part is the screening of the antibiotic susceptibility of Gram-negative bacteria (GNB) based the changes in their DEP behaviors. The -lactam antibiotics, whose function is to inhibit the cell wall synthesis, were used as the sample drugs to treat GNB. The -lactam-induced cell elongation happens in drug susceptible bacteria, and the significant change in DEP behavior can be observed at the inhibitory concentration within two hours. The antibiotic susceptibilities of important clinical pathogens (Escherichia coli and Klebsella pneumoniae) to -lactam antibiotics were tested by dielectrophoresis-based antibiotic susceptibility test (d-AST), and compared with the results of the standard culture method. In summary, the techniques of electrokinetics and microfluidics were integrated into one device for rapidly detecting bacteria and their drug sensitivity. Most importantly, the time needed of bacteria growth can be reduced from days to hours, and the occurrence of multidrug resistant bacteria also can be avoided. Ⅲ. AC electrokinetic motions of colloidal particles on the electrically-induced wave structure In the last subject, we demonstrated the electrically-assisted lithography that the wave structures induced by non-uniform electric fields were fabricated on the interdigitated electrode (IDE) arrays, and then colloidal particles were manipulated on the wave structure. In low frequency (~30 kHz), AC electroosmosis (ACEO) is predominant that particles were transported along valleys of the wave structure. The DEP force became stronger as the frequency raised to hundreds of kHz (~200 kHz), and then particles were collected at peaks of wave structures. Particles also can be concentrated at specific positions via changing the AC waveform. In addition, the size-dependant particle separation on the wave structure was also performed. Different size particles (0.5 and 2 m) can be separated into two populations due to the joint effect of drag force and DEP force, as they were transported by ACEO along the valley of the wave structure. The wave structures were fabricated rapidly and simply through the DEP-assisted lithography; and further, the joint AC electrokinetic phenomena of particles under the non-uniform electric fields were observed. The cell assembly, pattern, transportation and separation are potential applications to be going to achieve.

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