基于滑移芯片的双浓度梯度反应阵列  

Microfluidic Slipchip-based Reaction Microarray with Dual Concentration Gradient

作　　者：鄢兴华 胡亮[1] 李颖[1] 冯晓均[1] 刘笔锋[1] 

机构地区：[1]华中科技大学生命科学与技术学院,武汉430074

出　　处：《分析化学》2015年第10期1520-1525,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金资助项目(No.21475049;31471257;21275060)~~

摘　　要：建立了一种基于微流控滑移芯片的反应微阵列体系,并应用于双浓度梯度研究。在此反应微阵列芯片中,两种试剂分别通过芯片进样形成多级浓度梯度,进一步在正交方向上形成双浓度梯度的微反应阵列,经过滑动、接触和混合,简便实现多路反应。采用不同色素和荧光染料对系统进行了表征,确认了方法的可靠性,在此基础上,考察了β-半乳糖苷酶(β-gal)的反应条件。将45 nmol/Lβ-gal和45μmol/L底物荧光素二半乳糖苷(FDG)分别注入芯片上下层,在芯片通道中各自形成浓度梯度,滑移后进行不同酶浓度和不同底物浓度的反应测定。结果表明,随着底物浓度的提高,反应产物逐渐增多;一定底物浓度下,产物随酶浓度升高而逐渐增多,但很快到达平台期。芯片上酶与底物的浓度对反应的影响与芯片外基本一致。此芯片具有结构简单、操作简便和消耗低等特点,适用于多因素和多水平的复杂反应分析。Microarray technology has attracted increasing attentions in the field of bioanalysis,thanks to its advantages including high throughput and low sample and reagent consumptions. In this study,we developed a slipchip-based microfluidic system, in which sample and reagent could form two separate concentration gradients orthogonally. After sliding,samples and reagents of various concentrations could be mixed for multiplexed reactions. We validated the developed microfluidic system with food dyes and fluorescein. We further applied the method to investigate the reaction conditions involving β-galactosidase（ β-gal）. Solutions of45 nmol / L β-gal and 45 μmol / L FDG（ Fluorescein di-β-D-Galactoside） were injected into the upper layer and lower layer of the slipchip respectively. After the formation of two concentration gradients,one-step sliding of the upper layer enabled mixing and reacting of the enzyme and the substrate with different concentrations. As a result,the amount of the reaction products increased with the increasing concentrations of substrate. At a given concentration of substrate,the products rapidly reached a plateau with increasing concentrations of enzyme. These results were consistent with those obtained in 384-well plate. This method was easy to operate with low sample and reagent consumption, allowing rapid establishment of two orthogonal concentration gradients for multiplexed reactions.

关 键 词：微流控 滑移芯片 双浓度梯度 Β-半乳糖苷酶 

分 类 号：Q81[生物学—生物工程]