摘要
细胞融合是细胞工程最基本的核心技术之一,是细胞工程研究的重要内容。细胞融合技术已逐步从动物细胞的融合扩展到植物细胞、微生物细胞,甚至真菌的融合,导致了细胞工程技术的新突破。尽管细胞融合技术已经取得了长足进展,但仍然存在着诸多问题。针对目前正在使用的细胞融合技术的不足,本论文研究了一种微型化、可视化、集成化程度高、低成本、不会对细胞产生毒害作用、低电压驱动的细胞融合方法,并研究了与之密切相关的电介质电泳效应。这些研究工作对于发展细胞融合的相关理论和技术具有积极的作用,是在国内具有开拓性和创新性的工作。
本论文的主要研究内容如下:
(1)本论文应用细胞工程中的细胞融合理论、电介质物理理论、电场对细胞的作用理论,结合MEMS(Micro Electro Mechanical systems,微机电系统)技术,研究了一种新型的细胞融合方法。设计并制作完成一种基于MEMS技术和细胞电场效应的、可用于细胞融合的新型芯片——细胞电场效应融合芯片(Cell in Electrical-field Fusion Microchip,CEFM)。在理论分析和实验研究的基础上,并充分参考细胞融合电场诱导法中的相关技术参数,确定了芯片工作的电学条件。设计、制作完成驱动芯片工作的外围电路子系统,驱动细胞电场效应融合芯片正常工作。
(2)根据电介质物理理论和电场理论,在具体研究中特别注意了细胞的电场效应,研究了与CEFM密切相关的细胞电介质电泳效应,构建了细胞在CEFM中的电场效应模型(包括CEFM中悬浮液细胞的电学模型、CEFM中悬浮液细胞的电偶极子模型等)。并在理论分析基础之上,结合细胞电场效应融合芯片的研究进行了相关实验研究。相关工作成果指导了细胞融合芯片及其驱动电路的研制,这部分内容的理论分析与实验研究是CEFM相关研究的第一环节,也是CEFM相关研究中的基础研究环节。
本论文取得了以下主要研究结果:
(1)在关于细胞融合芯片中细胞排队方法的研究中,本论文先后研究了基于MEMS的微小室阵列和微电极阵列两种细胞融合芯片的设计思路和方案,并通过实验验证了微电极阵列细胞融合芯片的可行性;
(2)根据霍奇金-赫克利斯模型利用类比分析法构建起CEFM中悬浮液细胞的电学模型,从电介质物理Maxwell-Wagner的界面极化理论出发,构建了CEFM中悬浮液细胞的电偶极子模型。并进一步构建了CEFM细胞悬浮液中细胞的介电
The technology of cell fusion is one of elementary technologies of cellular engineering, and the basic method for the research of cellular engineering. Cell fusion technology has extended from the fusion of animal cell to the fusion of plant cell, the fusion of microbe, even the fusion of epiphyte. Absolutely, it results new breakthroughs for cellular engineering. However, there are many hampers, which embarrassed greatly cellular engineering more extensive and advanced development. As the case stands, to solve these crucial problems of cell fusion, a innovative method of cell fusion based on MEMS(Micro Electro Mechanical Systems)and the effect of cell in electric field was put forward. The research work is pioneer and innovative at home.
This dissertation accomplishes the research work as below:
(1) The theory of cell fusion, dielectric physics, the effect of cell in electric field, and MEMS technology have been concerned to develop a newly method used for cell fusion. A newly microchip, which is used to realize cell fusion and based on MEMS and effect of cell in electric field, have been designed and fabricated. Based on theoretical analysis, experimental investigation, and the data used in electrofusion, the technical parameters of circuit have been made certain. And utilizing the technique of software and electronics, the circuit used to driving the CEFM work well has been designed and made.
(2) According to the theory of dielectric physics and electric field, especially the effect of cell in electric field, dielectrophoresis, which associated with the CEFM closely, was investigated. The model of the effect of cell in the CEFM was set up, which includes the electric model of suspension cell in the CEFM, the dipole model of the suspension cell in the CEFM. Based on theoretical analysis and research works about the CEFM, experiments were made carefully. The electrical parameters about driving the microchip have been figured out. The work directed greatly the research of the cell fusion chip and the driving circuit. Theoretical investigation of dielectrophoresis about the CEFM has been analyzed carefully. This part of investigation is not only the first step but also the keystone of the CEFM research.
The results of the thesis could be drawn out as follows:
(1) Studying about the alignment of cell in the CEFM, the design scheme of the
引文
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