超支化聚酰胺酯改性聚甲基丙烯酸甲酯微流控芯片的制备及其在生物分子分离检测中的应用

利用亲水性超支化聚酰胺酯通过化学键合的方法对聚甲基丙烯酸甲酯(PMMA)微流控芯片的表面进行改性。对改性后PMMA微流控芯片的表面进行了接触角的测定,利用扫描电子显微镜(SEM)和体视显微镜观察了改性后芯片的表面形貌。结果表明,改性后的PMMA微流控芯片表面形成了一层均匀、致密、连续的亲水性涂层,芯片表面的亲水性得到了明显提高,接触角由未改性时的89.9°降低到29.5°。改性后芯片的电渗流较之改性前明显降低。利用芯片对腺苷和L-赖氨酸两种生物分子进行了分离检测。两种生物分子实现了完全分离,所得到的检测峰峰形尖锐,分离清晰。对腺苷和L-赖氨酸的分离柱效(理论塔板数)分别高达8.44×104 塔板/m和9.82×104 塔板/m,分离度(Rs)达到5.31,均远远高于未改性的芯片。改性后的芯片具有良好的分离时间重现性。本研究为提高PMMA微流控芯片的亲水性和应用范围提供了一种新的有效方法。     

聚甲基丙烯酸甲酯电泳芯片电化学检测氨基酸

采用集成铜微电极的聚甲基丙烯酸甲酯(PMMA)电泳芯片,实现对氨基酸的分离与检测。考察了制备的铜微电极对氨基酸的催化活性以及检测电位和分离场强对分离效果的影响。在优化条件(30mmol／L NaOH缓冲溶液中,检测电位为0．7V(vs.Ag／AgCl),进样与分离电场强度均为50V／cm)下,用3cm长的通道实现了精氨酸(Arg)与亮氨酸(Leu)的分离。     

一种连接石英毛细管的聚甲基丙烯酸甲酯电泳芯片的制作及其应用

利用模拟仿真软件模拟了死体积对于电泳分离的影响,并提出了一种实现聚甲基丙烯酸甲酯(Poly-methylmethacrylate,PMMA)电泳芯片与毛细管的最小死体积连接方法.将紫外吸收检测的检测窗口设计在石英毛细管上,使该PMMA芯片上的电泳分析可以直接采用紫外吸收检测器进行检测.利用该芯片对维生素B2进行了电泳分析,理论塔板数为73000/m;联磺甲氧苄啶片中的3种组分的分离度为4.5和1.9,3个峰迁移时间的RSD依次为1.0%,1.4%和1.0%,峰高的RSD依次为4.1%,3.0%和4.1%(n=5).     

通用型激光诱导荧光微流控芯片分析仪的研制与性能考察

设计和研制了一种通用型激光诱导荧光微流控芯片分析仪.检测部分按共聚焦检测原理设计,采用CCD(电荷耦合器件)监测通道,三维自动调节聚焦,发射波长滤光片可方便地更换以适应多种染料选择,能分别显示进样和分离通道2条电流-时间曲线.考察了该分析仪的检测灵敏度、检测极限和线性范围,显示了分析灵敏度高,检测限低和线性范围宽等特点,在自制注塑型PMMA塑料芯片上实现了φX174Haedi-gesT_dNA片段的分离测定和烟叶act基因PCR产物的分析     

聚甲基丙烯酸甲酯微流控分析芯片的简易热压制作法

提出聚甲基丙烯酸甲酯(PMMA)微流控分析芯片的一种简易热压制作法,研究了镍基、单晶硅和玻璃3种阳模制备芯片及芯片的封合条件.采用扫描电镜(SEM)和电荷耦合检测器(CCD)对PMMA芯片的微通道及其横截面形貌进行了表征.SEM图和CCD图表明实现了热压封接.测定了PMMA芯片的伏安曲线和电渗流,其电渗流值与文献报道值基本一致.本法制作的PMMA芯片用于电泳分离Cy5荧光染料,峰高RSD为2.2%(n=11),理论塔板数7.4×10⁴m^-1.     

基于智能手机的纸微流控电化学农药检测芯片的研究

纸质微流控装置的出现为低成本化学分析提供了一种简单而实用的平台.本研究开发了一种基于浓差电池原理的新型电化学检测纸质芯片,通过智能手机的辅助实现了农药的检测.检测芯片由色谱纸喷蜡打印制作而成.加入样品与芯片上的预加试剂反应5 min,然后将丝网印刷的电极层置于芯片上,利用模具的重力作用使电极层与纸芯片的两极紧密接触,再通过智能手机的USB读取装置获取芯片的电位,并由电位-农药浓度关系得到检测结果.使用此芯片实现了农药敌百虫(三氯磷酸酯)的快速、简便、可自供电的电化学定量检测,检出限为0.89 μ mol/L.     

功能型微流控芯片实验室的高通量和规模集成

以单一十字通道为基本构成,毛细管电泳为主体性能的第一代微流控芯片,曾经活跃了很长一段时间,至今仍是很大一部分同类芯片中的主流技术.我们曾在自行设计研制的波长为532 nm的激光诱导荧光电泳芯片仪上,分别采用自行设计研制的玻璃芯片、PMMA和PDMS塑料芯片,开展了DNA、蛋白质、糖蛋白的分离和手性药物的拆分研究,并完成了SARS病毒基因反转录多重PCR检测,癌症病人P16基因甲基化DNA诊断和髙血压基因筛査等的研究和相应的一定规模(分别为18例、159例和226例)的实际样品的测试,实现了微流控芯片系统的初级功能化.     

流体驱动智能电动控制仪用于PMMA微流控芯片电渗流的测量

采用自制的流体驱动智能电动控制仪作为电渗驱动控制源,结合电流跟踪法测量聚甲基丙烯酸甲酯(PMMA)芯片的电渗流,讨论了PMMA形成电渗的机理,给出了PMMA芯片通道电场强度与电渗速度的关系曲线。     

塑料芯片毛细管电泳电化学检测系统及其性能评价

近年来,高分子芯片毛细管电泳技术发展迅速,以聚甲基丙烯酸甲酯(PMMA)为代表的塑料电泳芯片由于其低廉的制作成本与良好的电渗性能,已经成为芯片电泳技术发展的一个重要方向,电化学检测具有灵敏度高、选择性好和易于微型化等优点,因此在塑料芯片电泳领域中具有较好的应用前景。     

CO2激光直写加工聚合物微流体芯片的建模研究

在CO2激光直写聚合物(PMMA)微流体芯片加工原理基础上,对激光直写PMMA微流道的数学模型进行了研究.采用能量守恒原理对激光直写PMMA微流道成型进行分析,结果表明,其微流道的形状呈高斯函数型分布,并获得了微流道深度和激光功率、加工速度的函数关系及微流道截面形状的数学模型.实验数据较好地验证了所建模型的正确性,该模型对CO2激光直写聚合物(PMMA)微流体芯片的加工工艺具有很好的指导作用.     

Influence of soluble polymer polyvinylpyrrolidone on separation of small peptides and amino acids by microchip-based capillary electrophoresis

In microchip-based capillary electrophoresis, the resolution and separation efficiency of small peptides and amino acids can be noticeably improved by adding a low molecular weight (30,000) soluble polymer additive, polyvinypyrrolidone in the separation medium. Several separation conditions such as injection time and electrophoretic buffer have been investigated and optimized. Using an electro-stacking scheme, the resolution and separation efficiency of small peptides and amino acids can be enhanced significantly. Under the optimal conditions, the separation of fluorescein isothiocyanate Isomer I-labeled small peptides and amino acids was successfully achieved within 100 s.     

Analysis of multiplex PCR fragments with PMMA microchip

Microchip electrophoresis is a promising technique for analysis of bio-molecules. It has the advantages of fast analysis, high sensitivity, high resolution and low-cost of samples. Plastic chip has the potential of mass production for clinical use for its advantages in biocompatibility and low cost. In this work, the method for fabrication of poly(methyl methacrylate) (PMMA) chip was described, and conditions for DNA separation were investigated with the chip. The PMMA microchip was used for detection of multiplex PCR products of 18 and 36 cases with SARS and hepatitis B virus infection under optimized separation conditions. Microchip electrophoresis showed higher sensitivity, higher resolution and less time consumption when compared with gel electrophoresis. The microchip electrophoresis with PMMA chip provided a rapid, sensitive and reliable method for analysis of multiplex PCR products.     

毛细管电泳微芯片在临床尿蛋白检测中的应用研究

用微芯片毛细管电泳法对临床患者尿蛋白进行了分离, 初步探讨了用于判断肾损伤的应用前景. 以pH 10.3, 75 mmol•L^－1的硼酸盐缓冲液作为芯片电泳缓冲体系, 利用蛋白质的紫外吸收特性, 在210 nm波段检测吸光度并进行信号收集和分析. 研究两种添加剂对提高尿蛋白分离效率的影响, 分析了肾病综合症、妊娠高血压症、风湿性心脏病和多发性骨髓瘤等患者尿样本, 并与美国Helena电泳系统分析结果对比, 得到了较一致的结果.     

Is capillary electrophoresis on microchip devices able to genotype uridine diphosphate glucuronosyltransferase 1A1 TATA‐box polymorphisms?

In this commentary, we focused our attention on capillary electrophoresis. It achieves the efficient separation of molecular species by the application of high voltages to samples in solution. Actually, capillary electrophoresis can be performed on microchip devices, based on an automated and miniaturized electrophoresis system, based on lab‐on‐a‐chip technology. By this technology it is possible to separate nucleic acid fragments (DNA or RNA) with respect to sizing accuracy and sizing resolution. Currently, two automated capillary electrophoresis on microchips devices are available: the Agilent 2100 Bioanalyzer and the Experion? Automated Electrophoresis System. In this study, we evaluated if the CE is able to distinguish the three uridine diphosphate glucuronosyltransferase 1A1 TATA‐box genotypes.     

High-speed separation of proteins by sodium dodecyl sulfate-capillary gel electrophoresis with partial translational spontaneous sample injection

In this study, we developed a picoliter-scale partial translational spontaneous injection approach which is suitable for high-speed protein separation under sodium dodecyl sulfate-capillary gel electrophoresis mode. On the basis of this approach, we built a high-speed CE system for protein separation based on a short capillary and slotted-vial array. The system has the advantages of simple structure, ease of building without the requirement of microfabricated devices, convenient operation, and low cost. Under the optimized conditions, picoliter-scale sample plugs (corresponding to ～65?μm plug length) were obtained, which ensured both the high speed and the high efficiency in protein separation. Five fluorescein isothiocyanate labeled proteins including myoglobin, egg albumin, bovine serum albumin, phosphorylase b, and myosin were separated within 60?s with an effective separation length of 1.5?cm. Theoretical plates per meter ranging from 2.58×10? to 1.28×10? (corresponding to 0.78-3.88?μm plate height) were obtained. The separation speed and separation efficiency of the present system are comparable to those of most microchip-based capillary electrophoresis systems for protein separation. The relative standard deviations of the migration times were in the range of 0.9-1.3% (n=5). Good linear relationships between log relative molecular mass and migration time were obtained in the molecular weigh range of 17,200-500,000, which demonstrate the present system can be applied in protein relative molecular mass determination.     

微流控芯片电泳分离血清中小而密低密度脂蛋白的研究

应用微流控芯片电泳,以40 mmol/L Tricine(pH9.8)作为电泳缓冲体系,十二烷基硫酸钠(SDS)作为添加剂(0.1 mmol/L SDS样品溶液,0.02 mmol/L SDS分离缓冲液),分离血清小而密低密度脂蛋白(sdLDL)。研究荧光染料硝基苯并噁二唑-C6-酰基鞘胺醇(NBD C6-ceramide)与脂蛋白结合的特异性、饱和性以及血清保存和检测时间对脂蛋白电泳行为的影响;探讨SDS有效降低蛋白吸附,提高血清脂蛋白分辨率的作用。冠心病(CHD)组sdLDL检出率(75%)显著高于对照组(6%,P<0.01)。该法具有简易、快速、高效等优点,可望成为CHD危险性评估的常规分析手段。     

Integrated circuit microchip system with multiplex capillary electrophoresis module for DNA analysis

In this paper, we describe the use of an integrated circuit (IC) microchip system as a detector in multiplex capillary electrophoresis (CE). This combination of multiplex capillary gel electrophoresis and the IC microchip technology represents a novel approach to DNA analysis on the microchip platform. Separation of DNA ladders using a multiplex CE microsystem of four capillaries was monitored simultaneously using the IC microchip system. The IC microchip-CE system has advantages such as low cost, rapid analysis, compactness, and multiplex capability, and has great potential as an alternative system to conventional capillary array gel electrophoresis systems based on charge-coupled device (CCD) detection.     

Quantitative analysis by microchip capillary electrophoresis: current limitations and problem-solving strategies

Obstacles and possible solutions for the application of microchip capillary electrophoresis in quantitative analysis are described and critically discussed. Differences between the phenomena occurring during conventional capillary electrophoresis and microchip-based capillary electrophoresis are pointed out, with particular focus on electrolysis, bubble formation, clogging, surface interactions, injection and aspects related to the power supply. Current drawbacks are specified and improvements for successful quantitative microchip capillary electrophoresis are suggested.     

卤代乙酸及其结构相近化合物的高效毛细管电泳分离

氟、氯、溴等卤代乙酸是结构非常相近的离子型化合物,对它们的分离测定比较困难。用高效毛细管电泳法在碱性或酸性缓冲液条件下可将它们分离。在酸性缓冲液条件下,可提高有机酸分离的选择性。较低的操作电压有利于提高阴离子的分离度,而改变温度对分离度的影响不大。     

Towards an integrated microfluidic device for spaceflight clinical diagnostics Microchip-based solid-phase extraction of hydroxyl radical markers

A microchip-based solid-phase extraction method for biological fluid small molecule analysis has been developed. Using a commercially available copolymer packed into a microchip channel, extraction and preconcentration of 2,3-dihydroxybenzoic acid (DHBA) and 2,5-DHBA from saliva was achieved. The metabolites, formed from salicylic acid by reactive oxygen species, can be used as markers of oxidative stress. The results show high recovery of both metabolites (>90+/-15% for spiked saliva) with an 80-fold concentration enhancement possible. The eluent is directly analyzed using capillary electrophoresis, with good resolution for the two metabolites. This study demonstrates the feasibility of future integrated microdevices for spaceflight small molecule biomarker analysis.