Design And Fabrication Of Arrayed High-throughput Cell Culture Chip

Microarray analysis technology is a novel and rapidly developing analytical method in recent years. It is an integrated product of multidiscipline knowledge such as micro-electro-mechanical system (MEMS), biochemistry, analytical chemistry, computer science, and so on.This thesis is mainly focused on the principle, methods and applications of the fabrication of microfluidic chip and the design, fabrication and optimization of a high-throughput cell-culture chip. The main contents include:1. The fabricating technique of the microfluidic chip2. The research and application progress microfluidic chip based high-throughput cell culture3. The design of an array based high-throughput cell-culture chip4. Experimental validation and chip structure improvementThe completed tasks mainly include:1. Describe the material choice and fabricating technique for different materials in the fabrication of microfluidic chip; introduce the improvement of the fabricating techniques on some kinds of materials did by our laboratory and the quartz, glass and PDMS chips fabricated by using improved technique by myself; summarize the applications of microfluidic chips in high-throughput cell culture, traveling-wave electrophoresis, and high-throughput drug screening.2. Present the design of the high-throughput cell-culture microfluidic chip: there are cell-culture chamber with U-style dam and tile-like mixing microchannels which are both based on tri-layer structures. SU8 photo-resist and PDMS were used as chip materials and"multi-photolithography and eventual development"process flow was proposed. Meanwhile, finite-element analytical software COMSOL, which could be used in couple of multiple physical fields, was introduced to simulate velocity field and the coupling of velocity field, convection and diffusion field in micro-chamber and mixing channel. Using the simulation results, the structures of cell culture chip was optimized.3. According to the experimental design, four microstructures including injector, medium mixed channels, cell-culture chamber, valve control structure, were designed and fabricated. Many instruments such as contact angle instrument, SEM, microplate reader were utilized to examine the contact angle, sizes of each microchannel and mixing efficiency. They were also used for the evaluation of the intercepting efficiency of the chamber for cells.Based on the systematical description of the fabricating principle and method of microfluidic chip, the array based cell-culture chip system for high-throughput cell culture was deeply studied. A set of feasible design, component fabrication, and verifying method was also set up.The study in this thesis could provide a credible platform for other application in microfluidic cell-culture chip based high-throughput manipulation, for example, culture and observe fused cells after high-throughput cell electro-fusion; culture and observe separated cells by high-throughput traveling-wave electrophoresis, and detect high-throughput cell by using laser confocal microscope. The microfluidic cell-culture chip designed and fabricated in this thesis has been validated repeatedly to be of good repeatability and consistency. Thus, it has established a solid foundation for the wide application of the microfluidic cell-culture chip.