【作者】 吕君江；

【导师】 徐溢；

【作者基本信息】 重庆大学 ， 药物化学， 2006， 硕士

【摘要】 本文将微流控芯片技术的高度集成能力、高效快速的分离能力、强大的流体操纵能力与酶免疫反应的高特异性及化学发光检测的高灵敏度相结合,研究了一种基于微流控芯片技术的血清地高辛临床检测方法。根据电泳芯片针对性强的特点,从芯片电泳基础理论出发,通过计算机模拟考察芯片电泳过程的电场及流场的分布,根据芯片电泳实验结果考察化学因素(化学反应、吸附、溶剂汽化)对芯片电泳过程的影响,对模拟计算结果进行验证,建立以样品性质和检测原理为起始参数的计算机模拟辅助电泳芯片的设计方法。针对目前需要常规检测的治疗窗口狭窄的强心甙类药物地高辛,提出血清地高辛酶免疫化学发光检测电泳芯片的整体设计方案,根据设计制作出电泳芯片,建立血清地高辛酶免疫化学发光芯片电泳检测方法,以期用于血清地高辛的临床检测。具体研究内容及相关结论包括:采用ConventorWare软件的微流体模块,对不同构型微混合管道的被动混合过程进行模拟计算和罗丹名6G发光成相实验,通过三维流场模拟图象及电场分布图象结合实验所得图象的分析,以混合效率为指标,讨论了不同结构微管道中层流混合过程的混合效果;对各混合微管道中罗丹明6G样品与缓冲液混合过程的混合效率进行计算,根据混合效率-混合长度曲线,论证和对比了引入不同构型的微管道混合方式下的混合效果。并根据模拟计算提出了一种结构和操作简单,高效而且可以用于电渗驱动混合的多级层流混合器的设计,用于血清地高辛电泳芯片的柱前免疫反应器。选择L-精氨酸和L-苯丙氨酸为分离样品体系,对十字形管道简单进样、夹流进样过程的电场分布和流场分布进行了模拟计算。通过讨论用于芯片电泳过程模拟的数学模型及其简化和求解方法,用十字形微管道实体模型模拟并优化了简单进样过程的进样时间和回流电压,得出简单进样需要有足够的充样时间使样品节的分布达到稳定,以反映样品本体溶液的浓度;通过模拟计算得到的三维图象以及样品浓度分布曲线和Cmax/σ-聚焦比曲线,综合分析得到夹流进样的聚焦比率应该在1.1-1.6范围内。根据电泳实验提出氨基酸样品基本参数,通过模拟计算考察了进样管道宽度和进样时间对进样方差的贡献;根据分离度与分离长度拟合曲线来确定电泳芯片的有效分离长度;对化学发光柱后衍生管道施加的夹流电压进行了模拟优化,得出氨基酸体系分离分析的电泳芯片设计方案和操作参数为:进样管道宽度为分离管道宽度的1/2,简单进样充样时间应大于5s,分离管道有效分离长度为30mm,衍生夹流比1.0-1.6。根据模拟优化结果提出的电泳芯片设计方案,采更多还原

【Abstract】 This work concerned to combine the highly integration, powerful separation, and skillful fluid manipulation of electrophoresis microchip with the specifity of immunoassay and the high sensitivity of chemiluminescence detection to develop a clinical detection method of serum digoxin based on microfluidic chips. Mathematical model of electrophoresis on microchip was taken to calculate and simulate the electric and flow field distributions in microchannel,which was validated by experimens. Electrophoresis and fluorescence photograming experiments on chip were conducted to investigate the effects of chemical reaction, reagents adsorption, and solutes volatilization on electrophoresis process, of which the results could not be obtained through simulation only. Combining the computer simulation with relative experiments, the computer assisted chip design procedure based on sample properties and detection principle was established. Focusing on the cardiac glycoside digoxin which has narrow therapeutic range and requires frequent monitoring, the chip applied to chemiluminescent enzyme immunoassay of serum digoxin was designed by the established design approach. The pre-column enzyme immuno reaction- electrophoresis separation- post-column chemiluminescence detection method of digoxin was developed, with the expectation to apply to clinical assay of digoxin. The research details and relative conclusions were inlcluding:ConventorWare, was employed to simulate the passive mixing processes within micro channels of different dimensions and structures. The simulation results were compared with the experimental images of Rhodamine 6G mixing process in glass-PDMS micro channels. According to the fluidic and electric fields distributions, mixing effects of laminar flow in micro channels of different geometric pattern were compared and discussed basing on the calculated mixing efficiency. The mixing process of Rhodamine 6G with buffer was performed and its mixing efficiency was calculated using MATLAB. In terms of the mixing efficiency versus mixing length curves, the contribution of different dimensional turns to mixing capability was quantified and validated. Furthermore, a new electro kinetic multiple laminar mixer with high performance was presented and validated by simulation and experiments, which only needed simpler fabrication and operation compared to the traditional one, which was applied to the pre-column immuno reactor of digoxin microchip.更多还原