本文選題：介電泳 + 流體動(dòng)力學(xué)��； 參考：《中北大學(xué)》2017年碩士論文

【摘要】：單細(xì)胞分辨率下細(xì)胞生物化學(xué)、物理特性的研究與表征,能夠有效揭示個(gè)體細(xì)胞在結(jié)構(gòu)功能與狀態(tài)上的差異性,對(duì)理解不同細(xì)胞新陳代謝、細(xì)胞性疾病的病理特征、細(xì)胞間的相互作用、細(xì)胞與介質(zhì)環(huán)境的相互作用等具有十分重要的意義。作為研究單細(xì)胞的重要手段,基于微流控芯片的單細(xì)胞排列與控制技術(shù)受到了愈來愈多的關(guān)注,然而目前大多數(shù)的單細(xì)胞排列與控制技術(shù)不僅成本高昂、而且效率很低,嚴(yán)重影響了單細(xì)胞排列與控制技術(shù)的推廣與應(yīng)用。本文結(jié)合介電泳的非侵入、免標(biāo)記、易控制與流體動(dòng)力學(xué)的高效率、低成本,通過仿真優(yōu)化和實(shí)驗(yàn)測(cè)試,設(shè)計(jì)制作了三明治式流體增強(qiáng)介電泳單細(xì)胞排列與控制芯片,實(shí)現(xiàn)了高通量陣列化的單細(xì)胞排列與控制,主要內(nèi)容如下:首先,對(duì)介電泳、絕緣介電泳和流體動(dòng)力學(xué)的基本理論做了分析,得出了介電泳力的公式及影響其大小與方向的因素、絕緣微墻對(duì)空間非均勻電場(chǎng)分布的影響及微墻對(duì)層流的影響;然后建立了等效細(xì)胞多層球殼模型,對(duì)細(xì)胞進(jìn)行受力分析得到流體增強(qiáng)介電泳捕獲與釋放細(xì)胞的原理;最后對(duì)OCI細(xì)胞的頻響特性進(jìn)行計(jì)算得出了不同電導(dǎo)率、不同介電常數(shù)溶液對(duì)細(xì)胞介電響應(yīng)的影響,為單細(xì)胞排列與控制奠定了理論基礎(chǔ)。其次,根據(jù)介電泳與流體動(dòng)力學(xué)的基本理論,利用計(jì)算機(jī)輔助軟件AutoCAD、Solidworks建立了三明治式流體增強(qiáng)介電泳單細(xì)胞排列與控制芯片的結(jié)構(gòu)并提出了其兩種工作模式,之后使用有限元軟件COMSOL Multiphysics仿真分析了芯片內(nèi)部電場(chǎng)和流場(chǎng)的分布,并通過仿真不同尺寸結(jié)構(gòu)對(duì)電場(chǎng)和流場(chǎng)的影響,得出了最佳的結(jié)構(gòu)尺寸。再次,根據(jù)微流控芯片選材原則,選定了各部分的加工材料與工藝,并根據(jù)加工方案,采用光刻法制作了上下層ITO微電極及SU-8微墻陣列,使用旋涂法制作了PDMS流體通道與導(dǎo)管接頭,最后使用氧等離子鍵合技術(shù)對(duì)芯片進(jìn)行了封裝。最后,搭建了微流控芯片實(shí)驗(yàn)測(cè)試平臺(tái),制作了用于連接芯片與外部信號(hào)源的PCB轉(zhuǎn)接板。培養(yǎng)人類急性髓細(xì)胞性白血病細(xì)胞(OCI-AML3),并用叉指電極優(yōu)化使OCI細(xì)胞發(fā)生介電泳行為的頻率與幅值后,通過熒光染色和處理OCI細(xì)胞,在制作的流體增強(qiáng)介電泳芯片中實(shí)現(xiàn)了OCI單細(xì)胞的陣列化捕獲與選擇性釋放,且單細(xì)胞捕獲效率大于95%。
[Abstract]:The study and characterization of the biochemical and physical properties of cells at single cell resolution can effectively reveal the differences in the structure, function and state of individual cells, and can be used to understand the pathological characteristics of different cell metabolism and cellular diseases.The interaction between cells and the interaction between cells and mediators is of great significance.As an important means to study single cell, the single cell arrangement and control technology based on microfluidic chip has been paid more and more attention. However, most of the single cell arrangement and control technology is not only expensive, but also inefficient.It seriously affects the popularization and application of single cell arrangement and control technology.In this paper, a sandwich fluid enhanced medium electrophoresis single cell array and control chip is designed and fabricated by simulation, optimization and experimental test, combining with the non invasive, label free, easy control and hydrodynamic high efficiency and low cost of medium electrophoresis.High throughput arrayed single cell alignment and control are realized. The main contents are as follows: firstly, the basic theories of dielectric electrophoresis, dielectric electrophoresis and hydrodynamics are analyzed.The formula of medium electrophoresis force and the factors influencing its size and direction, the influence of insulating microwall on spatial inhomogeneous electric field distribution and the effect of microwall on laminar flow are obtained, and the equivalent cell multilayer spherical shell model is established.The principle of fluid enhanced dielectric electrophoresis was obtained to capture and release the cells. Finally, the effects of different conductivity and dielectric constant solution on the dielectric response of OCI cells were calculated.It lays a theoretical foundation for the arrangement and control of single cell.Secondly, according to the basic theory of dielectric electrophoresis and fluid dynamics, the structure of sandwich fluid enhanced dielectric electrophoresis single cell arrangement and control chip was established by using computer aided software AutoCAD Solidworks, and two working modes were proposed.The distribution of electric field and flow field in the chip is simulated by using the finite element software COMSOL Multiphysics, and the optimum structure size is obtained by simulating the influence of different size structure on the electric field and flow field.Thirdly, according to the material selection principle of microfluidic chip, the machining materials and technology of each part are selected. According to the processing scheme, the upper and lower layer ITO microelectrode and SU-8 microwall array are fabricated by photolithography.The PDMS fluid channel and conduit joint were fabricated by spin-coating method. Finally, the chip was encapsulated by oxygen plasma bonding technology.Finally, the experimental test platform of microfluidic chip is built, and the PCB switch board is made to connect the chip with the external signal source.Human acute myeloid leukemia cells (OCI-AML3) were cultured, and the frequency and amplitude of dielectric electrophoresis behavior of OCI cells were optimized by cross finger electrode. The OCI cells were stained and treated by fluorescence staining.The single cell array capture and selective release of OCI were realized in the fluid enhanced dielectric electrophoresis chip, and the efficiency of single cell capture was greater than 95%.
【學(xué)位授予單位】：中北大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TN402

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相關(guān)期刊論文 前3條

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本文編號(hào)：1754489