生物细胞介电电泳运动控制机理及细胞排列生物芯片的研究
发布时间：2019-01-06 11:33
【摘要】：细胞操作技术对细胞的研究起到了重要的推动作用，介电电泳具有非接触、易控制优点，为细胞研究提供了一种无损操作方法。本文以介电电泳技术和微流控芯片为基础，分析了生物细胞介电电泳运动控制机理，设计了细胞阵列化排列生物芯片，并在实验上实现了细胞阵列化排列，对细胞的高通量检测和细胞间相互作用的研究具有一定的理论意义和应用价值。本文主要研究内容如下： 首先，对介电电泳的基本理论和细胞的介电电泳运动控制机理进行了分析。分析推导了交流电场中粒子的介电电泳力的公式，并研究了电场频率、溶液的电导率和介电常数对介电电泳的影响；然后根据细胞的特点，分析了多层式结构球体的介电电泳响应；最后分析了流体中的粒子受力情况，为细胞的介电电泳运动控制奠定了理论基础。 其次，根据介电电泳的原理，设计了细胞排列介电电泳芯片的整体结构，芯片为三层结构，顶部和底部均为电极层，中间为微通道。采用COMSOLMutiphysics仿真分析了芯片中的电势、电场强度及介电电泳的分布，并优化了芯片的结构参数，确定了电极的宽度为20μm，间距为40μm，顶部和底部电极的间距为50μm。 结合微流控芯片的特点，加工与封装介电电泳细胞排列芯片。介绍了ITO电极和PDMS通道的加工工艺过程，最后给出介电电泳细胞排列芯片的封装方法。 再次，，结合介电电泳芯片的特殊需求，搭建了介电电泳实验系统；然后对酵母菌细胞进行了正、负介电电泳操控实验，并讨论了交流电压的幅值、频率以及溶液电导率等因素对酵母菌细胞介电电泳的影响。实验发现，酵母菌细胞的介电电泳响应符合双层模型，随电场频率的增加，依次经历了负介电电泳、正介电电泳及负介电电泳三个阶段，确定细胞排列实验的交流电压频率为1MHz，幅值为8Vp-p，溶液的电导率为5μS/cm。 最后，在优化选取介电电泳操控参数下，应用加工的细胞排列芯片对酵母菌细胞进行了排列实验，酵母菌细胞受到正介电电泳力，向电场较强的区域运动，即顶部电极和底部电极的交叉点处，验证了芯片对细胞排列的可行性。
[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.
【学位授予单位】：中北大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TN492
本文编号：2402741
[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.
【学位授予单位】：中北大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TN492
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