集成电极的复合混沌混合芯片的快速制备及表征

试剂的有效混合是化学和生化反应的前提条件,因此混合成为微全分析系统的重要功能单元.本文介绍了一种通道中集成氧化铟锡(ITO)电极的聚二甲基硅氧烷(PDMS)-玻璃复合混沌混合芯片的快速制备方法,对PDMS的混合通道表面进行了硅溶胶改性.首次用集成的电极和酸碱反应过程中的电导变化对其混合效果进行了评价.利用玻璃各向同性刻蚀特点,一次曝光和一次刻蚀得到了混沌混合微通道结构的母板.利用光辅助原位聚合的方法快速制备了与母板微结构互补的聚甲基丙烯酸甲酯(PMMA)阳模,利用PI)MS原位聚合的方法复制得到了与玻璃母板结构相同的混沌混合通道结构的芯片.用电导法对芯片的混合有效性进行了表征.PDMS复制芯片与ITO玻璃永久封合即得全透明的复合混沌混合芯片.本文介绍的加工方法无需二次曝光及SU-8光刻胶,容易多次复制,在微流控分析芯片中将有广泛应用.     

用于PDMS微芯片塑性成型的SU-8模具制作工艺的优化

PDMS是制作微流控芯片的主要材料.PDMS芯片制作的主要方法是模塑法,模塑法要求有良好的塑性成型模具.SU-8以其良好的微加工特性,目前已广泛应用于微机械结构的制作,也用于PDMS塑性成型的模具.本文根据模具的特殊性,如平整、无裂纹、可多次使用等要求,研究了影响SU-8模具结构与基底材料硅片的黏附性和形成裂纹的因素,优化了SU-8微模具加工工艺,在以0.5℃/min进行升降温、210 mJ/cm2的曝光剂量、200℃条件下硬烘30min条件下得到较好的SU-8模具,提供了一种快速、复用性高、低成本的PDMS微芯片塑性成型的SU-8模具的制作方法.     

基于微流控芯片技术的小尺寸微米级液滴制备

利用微流控芯片技术制备的皮升级微量液滴,作为独立的微反应器,由于其比表面大,高通量等优势,在生物、医学、化学、物理等领域得到了广泛应用。本工作利用软光刻技术制备流聚焦型微流控芯片,研究了微流控芯片中连续相和分散相的流速对微量液滴尺寸的影响。结果显示,增加连续相流速时,微量液滴的尺寸减小;而加快分散相流速时,微量液滴的尺寸增大。当微流控芯片的通道尺寸固定后,由于分散相和连续相的界面张力不变,通过改变连续相和分散相的流速,微量液滴的尺寸范围有限,本工作中微量液滴的尺寸为几百微米至25μm。本工作探究了微流控芯片中如何制备尺寸小于25μm的微量液滴的方法。通过添加活性剂,改变连续相和分散相的界面张力,可实现制备尺寸为10μm的微量液滴。本工作所利用的微量液滴制备方法,制备的10μm大小液滴具备更高的比表面积,反应活性将会更大、在药物释放,颜色显示等领域将有广阔应用前景。     

塑料微流控芯片的注塑成型

有别于传统的微流控芯片压塑成型方法,本文提出注塑成型加工塑料微流控芯片的新工艺.采用UV-LIGA技术制作成型微通道的型芯,设计制造了微流控芯片注塑模具.充模试验表明,如何使微通道复制完全是微流控芯片注塑成型的主要技术难点.模拟与理论分析表明,熔体在微通道处出现滞流现象是复制不完全的主要原因;搭建了可视化装置对此加以试验验证.利用正交试验方法进行充模试验,研究各工艺参数对微通道复制度的影响.试验表明模具温度对提高微通道复制度起决定性作用;注射速度和熔体温度是次要因素,而注射压力相对其他因素影响力较差,但必须保持在一个较高的水平.依此形成塑料微流控芯片的注塑成型工艺,对于宽80μm、深50μm截面的微通道而言,可使微通道复制度由70%提高到90%,满足使用要求.     

毫米级水溶性芯材胶囊的微流控制备方法研究

为开发水溶性芯材胶囊制备新技术,基于微/毫流控技术设计了一种三维结构流动聚焦微通道芯片,采用该芯片制备了水包油包水(W/O/W)型双重乳液,以特种改性环氧丙烯酸酯为中间相,通过在线光固化方式进行中间相的固化,从而制备出粒径均一、颗粒圆整、包封率高的毫米级水溶性芯材胶囊,并研究了流速等因素对乳液形成过程、乳液粒径的影响。当三相流速分别为10mL/h、12mL/h、110mL/h时,所制备的微胶囊平均粒径为2.02mm,分散度小于3%。微/毫流控技术具有较好的稳定性和可操控性,实验装置具有易安装、易拆卸、便于清洗及可重复多次利用等优点。选用绿色环保、高效的光固化方式实现了胶囊壁的快速固化和高效封装。微/毫流控技术设备简单、容易操作,速度快、通量高,溶剂浪费少,达到节能降耗和环境保护的目的,适用于工业推广。     

新型纳米磁色谱微流体芯片

通过标准光刻电铸工艺,在玻璃基片上制备了不同形状的顺磁性镍铁微柱阵列作为微流控通道内的磁力元件;通过SU-8胶光刻模板PDMS快速成型制备了微流控通道结构,通过O2 Plasma 处理和显微镜下对准,实现在玻璃基片上的永久封装,制备了磁分离微流控芯片;在微流控通道中引入含有微磁珠的溶液,通过外加磁场和流动式进样,观察微磁珠的对磁场的响应情况以及层流中的磁珠分离情况及捕获,并进行了DNA提取实验.     

类荷叶表面疏水结构的材料表面制备

利用纳米/微米复模成型的方法,制备了荷叶表面微乳突状疏水结构的聚乙烯醇(PVA)和聚苯乙烯(PS)阴模模具,并利用阴模复模成型在聚二甲基硅烷(PDMS)表面制备了类荷叶的表面结构.扫描电子显微镜(SEM)的观察表明PDMS材料表面上制得的类荷叶结构与荷叶表面的微乳突结构有较好的一致性,而PVA阴模在保持微观结构上更有优势.通过对水滴在PDMS材料表面接触角的测量,证明了在制备有类荷叶表面结构的PDMS材料表面上,水滴的接触角可以得到显著提高.     

聚合物微流控芯片的注射成型

针对注塑成型微流控芯片过程中出现翘曲变形和微通道复制精度不高等缺陷,采用正交分析法,仿真优化了芯片厚度方向上的翘曲变形;基于翘曲优化结果,实验研究了微注射成型微流控芯片过程中模具温度、熔体温度和注射速度对微通道变形的影响。结果表明,保压时间和保压压力对微流控芯片的翘曲变形影响最大,而模具温度对微通道变形影响最为显著。采用优化的工艺参数,所成型的芯片微通道具有较高的复制度,无明显翘曲变形,可满足使用要求。     

PDMS微流控电泳芯片的快速制备

采用Protel软件绘制微流控沟道的形状,利用电路板制作技术加工出模具.该芯片由PDMS基片和PDMS盖片组成,微流控沟道位于基片上,深度和宽度分别为75μm和100μm,由盖片对其进行密封.考察了有绝缘漆模具和无绝缘漆模具制作的芯片的电泳分离情况.在所制作的PDMS微流控电泳芯片上对用异硫氰酸酯荧光素标记的氨基酸进行了电泳分离,当信噪比S/N=3时,最小检测浓度达到0.8×10-11mol/L.     

微流控芯片基片与盖片一体化注塑成型研究

针对微流控芯片基片与盖片的结构特点,提出了定模先行抽芯机构,设计制造了微流控芯片基片与盖片一体化注塑成型模具,并进行注塑成型试验研究.结果表明:定模先行抽芯机构可以有效解决盖片上圆孔状储液池成型与脱模的技术难题,如何使微通道复制完全是微流控芯片基片注塑成型的主要技术难点;模具温度对提高微通道复制度起决定性作用,注射速度和熔体温度是次要因素,而注射压力相对其他因素影响力较差,但必须保持在一个较高的水平,依此形成塑料微流控芯片的注塑成型工艺规范.     

不可逆封合微流控芯片中高活性蛋白质阵列的制备

保持生物分子的高活性是在不可逆封合微流控芯片中构筑微阵列芯片的关键问题.首先,利用MEMS技术和表面修饰方法制作了一种聚二甲基硅氧烷(PDMS)/玻璃芯片.应用光刻技术制作了PDMS盖片上的通道,同时用光刻剥离技术制作了玻璃基片上的金膜图案.进而,使用双官能团修饰剂3-氨丙基三甲氧基硅氧烷(APTMS)在玻璃基体和金膜图案上进行选择性表面修饰以吸附形成蛋白质阵列,并在其上覆盖一层水溶性聚乙烯醇(PVA)来保护蛋白质,既可避免其在加热处理过程中的高温伤害,又能防止在PDMS盖片与玻璃基片进行不可逆封合过程中的氧等离子体轰击造成的活性伤害.然后,通入水溶液冲洗除去PVA膜.使用荧光显微镜和原子力显微镜(AFM)考察蛋白质阵列质量,并结合免疫反应实验和细胞捕获固定实验评估蛋白质阵列的活性.结果表明,使用该方法可在不可逆封合的微流控芯片制作中构筑具有直径为200μm的高分辨率蛋白质阵列图案,蛋白质保持高的免疫活性,且可用于固定Hela细胞.     

Electrohydrodynamic generation and delivery of monodisperse picoliter droplets using a poly(dimethylsiloxane) microchip

We developed a drop-on-demand microdroplet generator for the discrete dispensing of biosamples into a bioanalytical unit. This disposable PDMS microfluidic device can generate monodisperse droplets of picoliter volume directly out of a plane sidewall of the microfluidic chip by an electrohydrodynamic mechanism. The droplet generation was accomplished without using either an inserted capillary or a monolithically built-in tip. The minimum droplet volume was approximately 4 pL, and the droplet generation was repeatable and stable for at least 30 min, with a typical variation of less than 2.0% of drop size. The Taylor cone, which is usually observed in electrospray, was suppressed by controlling the surface wetting property of the PDMS device as well as the surface tension of the sample liquids. A modification of the channel geometry right before the opening of the microchannel also enhanced the continuous droplet generation without applying any external pumping. A simple numerical simulation of the droplet generation verified the importance of controlling the surface wetting conditions for the droplet formation. Our microdroplet generator can be effectively applied to a direct interface of a microfluidic chip to a biosensing unit, such as AMS, MALDI-MS or protein microarray-type biochips.     

基于组织液超滤提取的MEMS血糖检测传感器

基于超滤原理提取组织液、并对其进行后续葡萄糖检测,是实现长期血糖持续监测的一种有效途径．本文提出一种可用于组织液超滤提取及葡萄糖持续检测的传感器微系统．该系统主要由微流控底座和葡萄糖传感器芯片组成．其中微流控底座由PDMS微通道、SU一8单向阀等微加工器件组成,在压力作用下可完成组织液提取及将检测过的组织液排出的功能．采用体硅加工方法制作葡萄糖传感器芯片微型腔体及腔体底部的微孔膜,研制出具有扩散控制功能的三电极检测芯片,并在其上通过琼脂糖包埋方法固定葡萄糖氧化酶、基于电化学原理实现葡萄糖浓度的检测．实验结果表明,该系统可以实现液体的灵活提取,并且葡萄糖检测响应时间小于5s,在0．4V工作电压下线性测量范围达0．2～20mmol／L,灵敏度为9．76nA／（mmol·L-1）,相关系数为0．9954．多次测量5mmoL／L样本,差异系数3．48％．可见该传感系统具有较好的稳定性,并且体积小、易于集成,有望用于组织液灵活提取及其葡萄糖持续监测．     

Addressable Microfluidic Polymer Chip for DNA‐Directed Immobilization of Oligonucleotide‐Tagged Compounds

A microfluidic polymer chip for the self‐assembly of DNA conjugates through DNA‐directed immobilization is developed. The chip is fabricated from two parts, one of which contains a microfluidic channel produced from poly(dimethylsiloxane) (PDMS) by replica‐casting technique using a mold prepared by photolithographic techniques. The microfluidic part is sealed by covalent bonding with a chemically activated glass slide containing a DNA oligonucleotide microarray. The dimension of the PDMS–glass microfluidic chip is equivalent to standard microscope slides (76 × 26 mm²). The DNA microarray surface inside the microfluidic channels is configured through conventional spotting, and the resulting DNA patches can be conveniently addressed with compounds containing complementary DNA tags. To demonstrate the utility of the addressable surface within the microfluidic channel, DNA‐directed immobilization (DDI) of DNA‐modified gold nanoparticles (AuNPs) and DNA‐conjugates of the enzymes glucose oxidase (GOx) and horseradish peroxidase (HRP) are carried out. DDI of AuNPs is used to demonstrate site selectivity and reversibility of the surface‐modification process. In the case of the DNA–enzyme conjugates, the patterned assembly of the two enzymes allows the establishment and investigation of the coupled reaction of GOx and HRP, with particular emphasis on surface coverage and lateral flow rates. The results demonstrate that this addressable chip is well suited for the generation of fluidically coupled multi‐enzyme microreactors.     

Poly(dimethylsiloxane)-based microchip for two-dimensional solid-phase extraction-capillary electrophoresis with an integrated electrospray emitter tip

A microchip in poly(dimethylsiloxane) (PDMS) for in-line solid-phase extraction-capillary electrophoresis-electrospray ionization-time-of-flight mass spectrometry (SPE-CE-ESI-TOF-MS) has been developed and evaluated. The chip was fabricated in a novel one-step procedure where mixed PDMS was cast over steel wires in a mold. The removed wires defined 50-microm cylindrical channels. Fused-silica capillaries were inserted into the structure in a tight fit connection. The inner walls of the inserted fused-silica capillaries and the PDMS microchip channels were modified with a positively charged polymer, PolyE-323. The chip was fabricated in a two-level cross design. The channel at the lower level was packed with 5-microm hyper-cross-linked polystyrene beads acting as a SPE medium used for desalting. The upper level channel acted as a CE channel and ended in an integrated emitter tip coated with conducting graphite powder to facilitate the electrical contact for sheathless ESI. An overpressure continuously provided fresh CE electrolyte independently of the flows in the different levels. Further studies were carried out in order to investigate the electrophoretic and flow rate properties of the chip. Finally, six-peptide mixtures, in different concentrations, dissolved in physiological salt solution was injected, desalted, separated, and sprayed into the mass spectrometer for analysis with a limit of detection in femtomole levels.     

基于转写技术的微针阵列制备与强度特性测试

为达到低成本、批量化制备微针阵列的目的,提出了一种分别制备微针针尖模具和微针立柱模具的微针模具制备方法.制备微针针尖硅模具是采用湿法刻蚀方法,SU-8微针的立柱部分则采用套刻工艺制备.以此模具为母版,采用聚二甲基硅氧烷(PDMS)二次转写技术获得PDMS二次母版.以PDMS母版为模具,分别用浇铸法制备了3种不同聚合物材料(左旋聚乳酸(PLLA),聚苯乙烯(PS),透明质酸(HA))的微针阵列;还以PDMS母版为模具,用电铸法制备了金属Ni微针阵列.该微针阵列的密度约为300根针/cm2.对制备的4种微针进行力学特性测试,实验结果表明,加工出的微针有足够的力学强度,可用于无痛注射.     

Chip-based bioassay using bacterial sensor strains immobilized in three-dimensional microfluidic network

A whole-cell bioassay has been performed using Escherichia coli sensor strains immobilized in a chip assembly, in which a silicon substrate is placed between two poly(dimethylsiloxane) (PDMS) substrates. Microchannels fabricated on the two separate PDMS layers are connected via perforated microwells on the silicon chip, and thus, a three-dimensional microfluidic network is constructed in the assembly. Bioluminescent sensor strains mixed with agarose are injected into the channels on one of the two PDMS layers and are immobilized in the microwells by gelation. Induction of the firefly luciferase gene expression in the sensor strains can be easily carried out by filling the channels on the other layer with sample solutions containing mutagen. Bioluminescence emissions from each well are detected after injection of luciferin/ATP mixtures into the channels. In this assay format using two multichannel layers and one microwell array chip, the interactions between various types of samples and strains can be monitored at each well on one assembly in a combinatorial fashion. Using several genotypes of the sensor strains or concentrations of mitomycin C in this format, the dependence of bioluminescence on these factors was obtained simultaneously in the single screening procedure. The present method could be a promising on-chip format for high-throughput whole-cell bioassays.     

Fabrication of topologically complex three-dimensional microfluidic systems in PDMS by rapid prototyping

This paper describes a procedure for making topologically complex three-dimensional microfluidic channel systems in poly(dimethylsiloxane) (PDMS). This procedure is called the "membrane sandwich" method to suggest the structure of the final system: a thin membrane having channel structures molded on each face (and with connections between the faces) sandwiched between two thicker, flat slabs that provide structural support. Two "masters" are fabricated by rapid prototyping using two-level photolithography and replica molding. They are aligned face to face, under pressure, with PDMS prepolymer between them. The PDMS is cured thermally. The masters have complementary alignment tracks, so registration is straightforward. The resulting, thin PDMS membrane can be transferred and sealed to another membrane or slab of PDMS by a sequence of steps in which the two masters are removed one at a time; these steps take place without distortion of the features. This method can fabricate a membrane containing a channel that crosses over and under itself, but does not intersect itself and, therefore, can be fabricated in the form of any knot. It follows that this method can generate topologically complex microfluidic systems; this capability is demonstrated by the fabrication of a "basketweave" structure. By filling the channels and removing the membrane, complex microstructures can be made. Stacking and sealing more than one membrane allows even more complicated geometries than are possible in one membrane. A square coiled channel that surrounds, but does not connect to, a straight channel illustrates this type of complexity.     

侧壁四电极电容耦合非接触电导检测器(英文)

研制了一种集成于硅基电泳芯片分离沟道末端侧壁的新型四电极电容耦合非接触电导检测器.研究了该电导检测器的等效模型,对等效电路模型中的参数进行了公式推导,并讨论了影响电导检测响应灵敏度的相关因素.采用深刻蚀及离子注入加工技术制得了用于电导检测的立体电极.制作了基于锁相放大原理的信号处理电路,对该电导检测的频率响应及灵敏度进行了测试分析.实验结果表明,当激励信号频率为300 kHz时,该电导检测器具有最佳线性度;不同浓度Na+溶液响应电压差值为5 mV;检测限达到10-8mol/L;且成功实现了Na+和Li+混合无机阳离子的电泳分离在线检测.     

微流控芯片表面修饰及在蛋白质富集中的应用

应用分子自组装技术，在SiO2表面衍生活泼醛基，在SiO2表面有125nm的衍生物，衍生的醛基和氨基发生共价反应而将入免疫球蛋白G固定在二氧化硅表面，抗原抗体反应显示固定的抗体有活性。应用微加工技术加工含交叉排列的椭圆形微柱阵列的微流控芯片，有效增加内表面积和流体接触机会，用同样修饰方法修饰微流控芯片内表面并固定人免疫球蛋白G，流经管道的相应抗抗体和其发生反应而被吸附在管道表面，实现对该抗抗体的亲和富集，富集后荧光密度增加15倍。表面修饰技术能实现蛋白质在二氧化硅表面的固定并保持其生物活性，结合微流控芯片能实现对相应蛋白质的微量富集。