导　　师： 杨宜民;章云

学科专业： 081101

授予学位： 博士

作　　者： ;

机构地区： 广东工业大学

摘　　要： 显微注射是生物工程的基因导入中常用的方法，它导入外源基因的效率高。采用该法导入的外源基因往往是在细胞期就能整合到受体染色体上，发育成的体细胞和生殖细胞一般都能整合上外源基因，使外源基因在受体细胞上能获得好的表达，因此它日益受到生物界科研人员的信赖。目前80％以上的转基因动物模型都是利用显微注射来构建完成的。 显微注射技术对胚胎发育有着关键性的影响，如注射剂量的不同将直接影响到胚胎体外发育的存活率乃至影响整个转基因动物的成功率。采用手动方式进行微注射量控制，存在效率低、准确性差以及受人为因素影响较大等不足，限制了这项技术的推广及应用。 目前国外各大生物仪器制造公司纷纷推出了性能较为先进的微注射量控制器，使注射量控制中的手工劳动的程度大大降低，实现了向半自动化控制的转化，但这些设备高昂的价格往往让国内的许多从事基因工程的单位和机构难以接受。为了追赶世界先进水平，同时也为了开发具有中国特色的显微注射设备，以推动我国基因工程的发展，本论文在深入调查了解国外先进产品，以及从理论及实验方面对DNA微流体的动力学特性进行研究的基础上，致力于微注射量控制器的设计与研制，以及显微视觉检测技术的研究，以便对其进行更有效、更精确的定量控制。 围绕上述内容，本文进行了以下几方面的研究工作： /(1/)查阅并分析了国内外大量的转基因微注射技术及操作设备和微流体控制方面的文献，在对DNA微流体的动力学特性理论研究的基础上，进行了微注射量的理论建模及实验研究。 /(2/)对微注射量控制器进行了系统、参数及容差等方面的方案设计，并在原有的基础上进行了改进，构� Microinjection is often used in Gene Introduction in the Biological Engineering. Through it, the exterior genes are often effectively introduced into the chromosome of the embryonic period's receptors, and then will be integrated so that when the receptorsomatic and germ cells grow up, they will have a good expression. Therefore, microinjection is attracting more and more attention of the researchers in the biological fields. By far, about 80/% or more transgenic animals are constructed through microinjection. The technique of microinjection is significant to embryo's development. For example, the different volume of microinjection will have direct effect on the in vitro survival rate of the embryos and even the successful rate of the transgenic animals. There are some shortcomings in the microinjection whose volume is controlled by manual, such as less effective, less precise and more sensitive to artificial factors, etc. All these keep the microinjection operation from being more popular and applicable. At present some volume controllers with high-qualities which can argely decrease the manual work of the microinjection volume control are put up with by several international biological companies. With the new devices, microinjection is turned into a half-automatic one. However, these devices are too expensive to many domestic research associations and societies of the gene engineering. On the purpose of catching up with the advanced level of the world, and developing microinjection devices of ourselves to promote the Chinese gene engineering, this paper is dedicated to the design and fabrication of the microinjection volume controller and the research on the detection of microinjection volume by micro-vision, based on looking into the advanced products mentioned above and researches on the dynamical characteristics of the DNA micro-fluid from both theoretical and experimental aspects. Several research works are done in this paper according to the topics mentioned above: 1

关 键 词： 转基因 显微注射 微注射量 气压电控式 温度梯度 效应 显微图像 特征提取

领　　域： [自动化与计算机技术] [自动化与计算机技术]