【摘要】：細(xì)胞操作技術(shù)對(duì)細(xì)胞的研究起到了重要的推動(dòng)作用，介電電泳具有非接觸、易控制優(yōu)點(diǎn)，為細(xì)胞研究提供了一種無(wú)損操作方法。本文以介電電泳技術(shù)和微流控芯片為基礎(chǔ)，分析了生物細(xì)胞介電電泳運(yùn)動(dòng)控制機(jī)理，設(shè)計(jì)了細(xì)胞陣列化排列生物芯片，并在實(shí)驗(yàn)上實(shí)現(xiàn)了細(xì)胞陣列化排列，對(duì)細(xì)胞的高通量檢測(cè)和細(xì)胞間相互作用的研究具有一定的理論意義和應(yīng)用價(jià)值。本文主要研究?jī)?nèi)容如下： 首先，對(duì)介電電泳的基本理論和細(xì)胞的介電電泳運(yùn)動(dòng)控制機(jī)理進(jìn)行了分析。分析推導(dǎo)了交流電場(chǎng)中粒子的介電電泳力的公式，并研究了電場(chǎng)頻率、溶液的電導(dǎo)率和介電常數(shù)對(duì)介電電泳的影響；然后根據(jù)細(xì)胞的特點(diǎn)，分析了多層式結(jié)構(gòu)球體的介電電泳響應(yīng)；最后分析了流體中的粒子受力情況，為細(xì)胞的介電電泳運(yùn)動(dòng)控制奠定了理論基礎(chǔ)。 其次，根據(jù)介電電泳的原理，設(shè)計(jì)了細(xì)胞排列介電電泳芯片的整體結(jié)構(gòu)，芯片為三層結(jié)構(gòu)，頂部和底部均為電極層，中間為微通道。采用COMSOLMutiphysics仿真分析了芯片中的電勢(shì)、電場(chǎng)強(qiáng)度及介電電泳的分布，并優(yōu)化了芯片的結(jié)構(gòu)參數(shù)，確定了電極的寬度為20μm，間距為40μm，頂部和底部電極的間距為50μm。 結(jié)合微流控芯片的特點(diǎn)，加工與封裝介電電泳細(xì)胞排列芯片。介紹了ITO電極和PDMS通道的加工工藝過(guò)程，最后給出介電電泳細(xì)胞排列芯片的封裝方法。 再次，，結(jié)合介電電泳芯片的特殊需求，搭建了介電電泳實(shí)驗(yàn)系統(tǒng)；然后對(duì)酵母菌細(xì)胞進(jìn)行了正、負(fù)介電電泳操控實(shí)驗(yàn)，并討論了交流電壓的幅值、頻率以及溶液電導(dǎo)率等因素對(duì)酵母菌細(xì)胞介電電泳的影響。實(shí)驗(yàn)發(fā)現(xiàn)，酵母菌細(xì)胞的介電電泳響應(yīng)符合雙層模型，隨電場(chǎng)頻率的增加，依次經(jīng)歷了負(fù)介電電泳、正介電電泳及負(fù)介電電泳三個(gè)階段，確定細(xì)胞排列實(shí)驗(yàn)的交流電壓頻率為1MHz，幅值為8Vp-p，溶液的電導(dǎo)率為5μS/cm。 最后，在優(yōu)化選取介電電泳操控參數(shù)下，應(yīng)用加工的細(xì)胞排列芯片對(duì)酵母菌細(xì)胞進(jìn)行了排列實(shí)驗(yàn)，酵母菌細(xì)胞受到正介電電泳力，向電場(chǎng)較強(qiáng)的區(qū)域運(yùn)動(dòng)，即頂部電極和底部電極的交叉點(diǎn)處，驗(yàn)證了芯片對(duì)細(xì)胞排列的可行性。
[Abstract]:Cell manipulation technology plays an important role in cell research. Dielectric electrophoresis has the advantages of non-contact and easy to control, which provides a non-destructive operation method for cell research. Based on dielectric electrophoretic technology and microfluidic chip, the motion control mechanism of dielectric electrophoretic of biological cells is analyzed, cell array biochip is designed, and cell array is realized experimentally. The study of cell high throughput detection and intercellular interaction has certain theoretical significance and application value. The main contents of this paper are as follows: firstly, the basic theory of dielectric electrophoresis and the mechanism of cell motion control are analyzed. The formula of dielectric electrophoretic force of particles in alternating current field is deduced, and the effects of electric field frequency, conductivity of solution and dielectric constant on dielectric electrophoresis are studied. Then, according to the characteristics of cells, the dielectric electrophoretic response of multilayer structure spheres is analyzed. Finally, the stress of particles in the fluid is analyzed, which lays a theoretical foundation for the control of dielectric electrophoresis motion of cells. Secondly, according to the principle of dielectric electrophoretic, the whole structure of cell array dielectric electrophoresis chip is designed. The chip is composed of three layers, the top and bottom are electrode layer, and the middle is microchannel. The distribution of electric potential, electric field intensity and dielectric electrophoresis in the chip was analyzed by COMSOLMutiphysics simulation. The structure parameters of the chip were optimized. The electrode width was 20 渭 m, the spacing was 40 渭 m, and the distance between the top and bottom electrodes was 50 渭 m. Based on the characteristics of microfluidic chips, the dielectric electrophoretic cell array chips were fabricated and packaged. The fabrication process of ITO electrode and PDMS channel is introduced. Finally, the encapsulation method of dielectric electrophoretic cell array chip is given. Thirdly, according to the special requirement of dielectric electrophoresis chip, a dielectric electrophoretic experiment system is built. Then the positive and negative dielectric electrophoretic manipulation experiments were carried out on yeast cells, and the effects of the amplitude of AC voltage, frequency and solution conductivity on the dielectric electrophoresis of yeast cells were discussed. The results showed that the dielectric electrophoretic response of yeast cells was in accordance with the double-layer model. With the increase of electric field frequency, the response of yeast cells went through three stages: negative dielectric electrophoresis, positive dielectric electrophoresis and negative dielectric electrophoresis. The AC voltage frequency of cell arrangement experiment is 1 MHz, the amplitude is 8 Vp-p, and the conductivity of the solution is 5 渭 S / cm ~ (-1). Finally, under the optimized selection of dielectric electrophoretic manipulation parameters, the yeast cells were arranged by the processed cell array chip. The yeast cells were subjected to positive dielectric electrophoretic force and moved to the region with strong electric field. The cross point of top electrode and bottom electrode verifies the feasibility of cell arrangement.
【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】：2015
【分類號(hào)】：TN492

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本文編號(hào)：2402741