微流体数字化细胞显微注射实验研究

【作者】 兰海英；

【导师】 侯丽雅；

【作者基本信息】 南京理工大学 ， 机械制造及其自动化， 2008， 硕士

【摘要】 生物技术的发展以及细胞注射技术的广泛应用,对实验手段和实验设备提出了越来越高的要求。比如,设备操作的容易程度、设备对微操作对象的影响大小、微注射量的控制精度等问题。在细胞注射过程中,对微操作要求迅速准确,注射速度快;细胞刺膜过程对细胞的变形影响小、微注射量一般要求达到pL级。目前还没有相关设备和手段来完全解决这些问题,南京理工大学微系统研究室提出的“微流体数字化技术”在这方面取得了突破性进展。本文是针对数字化细胞微注射仪的细胞注射实验研究,该注射仪是基于“微流体数字化技术”研发的。利用微系统研究室研发的细胞工程用微针制备仪器,在大量制备微针实验的基础上,制作出符合细胞注射要求的微针。对微针进行表面疏水化以及清洗处理,可以提高微注射针的注射性能;利用图像处理技术可以精确测出其物理参数。此外,对数字化微注射仪的进针、退针性能进行测试;重点研究了注射仪的注射性能,得到不同参数下的注射量,并对其影响因素进行分析。在此基础上,对三种具有代表性的卵细胞进行微注射操作,通过实验探索注射驱动参数,得出注射成功率较高的参数范围。最后对注射过荧光标记物的细胞进行荧光检测,证实微注射仪确实将注射药液注入细胞内。更多还原

【Abstract】 With the development of biotechnology and the wider application of cell injection technology, the necessity of qualified experimental means and facilities closes behind. For example, the facility of operation and fidelity of microinjection dose, and so on. Generally, quick and accurate micro operations, velocity of injection, small transfiguration of cell and microinjection dose to the degree of pL are required in executing cell injection. There are no good equipment and means to resolve these issues yet. So it’s very important to research on this respect. "Digitalization of micro fluids technology" from micro system lab of Nanjing University of Science and Technology has a breakthrough at this respect.This paper is for experimental research of cell injection by using digital micro-injection device which is based on "digitalization of micro fluids technology". The micro needle preparation devices which are used in Cell Engineering and researched by micro-system lab can create micro-injection needles. The micro-injection needles are fabricated by a large number of micro-needle preparation experiments. The performances of injecting droplets are enhanced after the surface hydrophobic and wash treatment. The micro-needles’ precise physical parameters are measured by image processing technology. Then, some tests are made to detect the performance about advance and retreat of digital micro-injection device. Its injection performance are researched to get different parameters of the injection, and then the factors of the injection quantity are analyzed. On this condition, author carry out micro injection experiment on three typical and different cells. Search after drive parameter of injection, and then get the parameter range of the high ration of successful injecting. Finally, confirmed that micro-injection device can inject drug into the cell by fluorescence detecting on the cell which is injected fluorescent marker.更多还原