超支化聚胺-酯改性聚二甲基硅氧烷微流控芯片的紫外检测应用

采用超支化聚胺-酯对经过氧气氛处理的PDMS微流控芯片表面进行改性.成功地将超支化聚胺-酯涂覆到PDMS表面,使其表面的接触角由108°±1°降到32°±20°,改善了其亲水性;改性过后通道内的电渗流得到了有效抑制,远低于未改性通道内的电渗流.同时,将芯片通过专门设计的通道与毛细管连接在一起,在紫外检测波长214nm,...     

聚二甲基硅氧烷微流控芯片的紫外光照射表面处理研究

研究了紫外光化学表面改性对聚二甲基硅氧烷(PDMS)微流控芯片的片基间粘接力及毛细管通道电渗流性能的影响.PDMS片基经紫外光射照后,粘接力增强,可实现PDMS芯片的永久性封合,同时亲水性得到改善,通道中的电渗流增大.与文献报道的等离子体表面处理方法比较,采用紫外光表面处理,设备简单,操作方便,耗费少,是一种简单易行的聚二甲基硅氧烷芯片表面处理方法.     

功能化PDMS芯片的制作及其在氨基酸电泳分离中的应用

PDMS芯片表面具有强疏水性,不仅使水溶液很难充满其管道,电渗流不易控制~([1～3]),而且由于生物分子在其表面的强烈吸附还会导致芯片受污染,这些问题影响了PDMS在微流控中的应用.因此,PDMS芯片表面修饰已成为微全分析研究的热点之一.     

异质材料微流体通道电渗流热效应

本文对玻璃和聚二甲基硅氧烷(PDMS)材料制作的微流控芯片电渗流焦耳热效应进行数值研究.采用双电层的Poisson-Boltzmann方程,液体运动的Navier-Stokes方程和液-固耦合系统的热传导方程研究二维微通道电渗流的温度特性.研究发现:由于材料属性的差别,温度场和速度场在微通道断面存在不均匀性.微通道表面的温升会降低双电层的电荷密度.热效应会对电渗流速度场产生影响,并诱导压强梯度和改变外电场在微通道的变化特征.     

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。     

聚甲基丙烯酸甲酯微流控芯片分离DNA片段的初步研究

自从1995年Mathies^[1]首次将微流控芯片毛细管凝胶电泳用于基因测序研究以来,DNA片段的分离已成为微流控芯片应用的重要领域之一.最早应用于DNA分析的微流控芯片是玻璃芯片,聚合物微流控芯片以其品种多、成本低、易于加工,与玻璃芯片相比具有封接温度大大降低,微通道内电渗流显著减小等特点,已被成功应用于DNA片段的分离^[2,3].     

激光诱导荧光微流控芯片分析仪的研制

研制了一种基于激光诱导荧光检测方法的微流控芯片分析仪。该分析仪使用玻璃基质聚二甲基硅氧烷(PDMS)微流控芯片,可一次性进行12通道的电泳分离实验。仪器采用共聚焦式光路结构,并可通过检测由微流控芯片反射的激光信息,控制步进电机实现芯片的自动精确定位。实验结束自动保存数据,绘制分离图谱。。对9种不同长度的50 bp DNA Ladder片段进行电泳分离及数据分析,耗时在5 min内,且分离效果良好。     

微流控芯片单细胞进样,溶膜及分离检测胞内谷胱甘肽

单细胞分析对重大疾病的早期诊断等方面有重要意义[1].微流控分析芯片的网络结构和微米级的通道尺寸适合于单细胞进样、溶膜和分离分析.但目前的报道主要集中在细胞培养、计数和筛选[2].我们在十字通道微流控芯片上,通过调节储液池的液面高度和细胞悬液密度,使单细胞逐个通过芯片进样通道和分离通道之间的区域,再结合控制电渗流方向,使单细胞固定在分离通指定位置,然后用电泳缓冲液结合高电场实现细胞快速溶膜,接着进行电泳分离和LIF检测.实现了单个血红细胞内谷胱甘肽(GSH)的高效分离及定量分析.     

聚二甲基硅氧烷芯片粘接强度的实验研究

聚二甲基硅氧烷(PDMS)材料广泛地应用于制作微流控芯片.本文研究了PDMS预聚体与固化剂的配比、固化温度和固化时间、固化模具以及紫外光照射等重要因素对PDMS芯片封接强度的影响,得到PDMS芯片封接的最佳条件为:基片和盖片所用PDMS预聚体与固化剂的最佳质量配比为10∶1,最佳固化温度为75℃,固化时间为40 min;采用不同材料模具制作PDMS片,其表面均方根粗糙度控制着芯片的粘接强度.在研究的三种模具材料中,用有机玻璃模具制作的PDMS片间的粘接强度最高,用玻璃模具制作的PDMS片间粘接强度最小;PDMS片经紫外光照射表面处理后,粘接强度会增加.     

聚对苯二甲酸乙二酯微流控芯片的制备和表面改性

高聚物微流控芯片具有制备简单、成本低、可批量生产等突出优点而有望成为一次性器件,近年来在国内外引起了同行们的兴趣.但是,高聚物表面的憎水性、对有机分子的强吸附性、电渗流的不稳定性等又成为高聚物芯片在微流控分析领域得以广泛应用的障碍.表面改性是改善高聚物芯片分析特性的有效途径.     

Multilayer poly(vinyl alcohol)-adsorbed coating on poly(dimethylsiloxane) microfluidic chips for biopolymer separation

A poly(dimethylsiloxane) (PDMS) microfluidic chip surface was modified by multilayer-adsorbed and heat-immobilized poly(vinyl alcohol) (PVA) after oxygen plasma treatment. The reflection absorption infrared spectrum (RAIRS) showed that 88% hydrolyzed PVA adsorbed more strongly than 100% hydrolyzed one on the oxygen plasma-pretreated PDMS surface, and they all had little adsorption on original PDMS surface. Repeating the coating procedure three times was found to produce the most robust and effective coating. PVA coating converted the original PDMS surface from a hydrophobic one into a hydrophilic surface, and suppressed electroosmotic flow (EOF) in the range of pH 3-11. More than 1,000,000 plates/m and baseline resolution were obtained for separation of fluorescently labeled basic proteins (lysozyme, ribonuclease B). Fluorescently labeled acidic proteins (bovine serum albumin, beta-lactoglobulin) and fragments of dsDNA phiX174 RF/HaeIII were also separated satisfactorily in the three-layer 88% PVA-coated PDMS microchip. Good separation of basic proteins was obtained for about 70 consecutive runs.     

Poly(oxyethylene) based surface coatings for poly(dimethylsiloxane) microchannels

Control of surface properties in microfluidic systems is an indispensable prerequisite for successful bioanalytical applications. Poly(dimethylsiloxane) (PDMS) microfluidic devices are hampered from unwanted adsorption of biomolecules and lack of methods to control electroosmotic flow (EOF). In this paper, we propose different strategies to coat PDMS surfaces with poly(oxyethylene) (POE) molecules of varying chain lengths. The native PDMS surface is pretreated by exposure to UV irradiation or to an oxygen plasma, and the covalent linkage of POE-silanes as well as physical adsorption of a triblock-copolymer (F108) are studied. Contact angle measurements and atomic force microscopy (AFM) imaging revealed homogeneous attachment of POE-silanes and F108 to the PDMS surfaces. In the case of F108, different adsorption mechanisms to hydrophilic and hydrophobic PDMS are discussed. Determination of the electroosmotic mobilities of these coatings in PDMS microchannels prove their use for electrokinetic applications in which EOF reduction is inevitable and protein adsorption has to be suppressed.     

Surface modification with BSA blocking based on in situ synthesized gold nanoparticles in poly(dimethylsiloxane) microchip

A stable BSA blocking poly(dimethylsiloxane) (PDMS) microchannel was prepared based on in situ synthesized PDMS–gold nanoparticles composite films. The modified microchip could successfully suppress protein adsorption. The assembly was followed by contact angle, charge-coupled device (CCD) imaging, electroosmotic flow (EOF) measurements and electrophoretic separation methods. Contact angle measurements revealed the coated surface was hydrophilic, water contact angle for coated chips was 45.2° compared to a water contact angle for native PDMS chips of 88.5°. The coated microchips exhibited reproducible and stable EOF behavior. With FITC-labeled myoglobin incubation in the coated channel, no fluorescence was observed with CCD image, and the protein exhibited good electrophoretic effect in the modified microchip.     

一种直接测定微流控芯片电渗流速度的新方法

随着微芯片技术的成熟,越来越迫切地需要有一个准确而简洁的电渗流速度的检测方法。根据荧光物质罗丹明123（Rh123）在不同pH缓冲溶液中迁移时间的变化,推导出Rh123在pH 9和10条件下分别有中性分子存在,而中性分子的移动速度等于电渗流速度,因此建立了直接以Rh123中性分子为标记物测定电渗流速度的方法。通过直接检测Rh123中性分子的迁移时间,计算得出所用玻璃微流控芯片在pH 9.3和pH 10.1的电渗流速度为3.9×10-4 cm2/(s·V)和4.1×10-4 cm2/(s·V),与经典方法对照无明显差异。     

Poly(dimethylsiloxane)-based microfluidic device with electrospray ionization-mass spectrometry interface for protein identification

An easy method to fabricate poly(dimethylsiloxane) (PDMS)-based microfluidic chips for protein identification by tandem mass spectrometry is presented. This microchip has typical electrophoretic microchannels, a flow-through sampling inlet, and a sheathless nanoelectrospray ionization (ESI) interface. The surface of the microchannel was modified with 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS) and the generated electroosmotic flow under acidic buffer condition used for the separation was found to be more stable compared to that generated by the microchannel without modification. The feasibility of the device for flow-through sampling, separation, and ESI-MS/MS analysis was demonstrated by the analysis of a standard mixture composed of three tryptic peptides. Results show that four peaks corresponding to three peptide standards and acetylated products of the standard peptide were well resolved and the deduced sequences were consistent with those expected. Furthermore, the compatibility of this device with other miniaturized devices to integrate the whole process was also explored by connecting a miniaturized enzymatic digestion cartridge and a desalting cartridge in series to the sampling inlet of the microchip for the identification of a model protein, beta-casein.     

Capillary zone electrophoresis of amino acids on a hybrid poly(dimethylsiloxane)-glass chip

Poly(dimethylsiloxane) (PDMS)-PDMS and hybrid PDMS-glass devices have been characterized and compared in terms of current-voltage linearity, contact angle, electroosmotic velocity, electroosmotic mobility, and electrokinetic potential in dependence on the surface treatment. The hybrid PDMS-glass microfluidic devices have further been tested as on-chip capillary electrophoresis systems for the separation of fluorescently labeled amino acids. It has been demonstrated that different methods of surface pretreatment of the PDMS-glass devices result in significantly different separation performance, with plate numbers varying from 650 to 57 000 in dependence on the surface state and the nature of the amino acids. Electrophoretic separations of amino acids have been achieved within tens of seconds with detection limits of less than 2 microM (approximately 2 x 10(-16) to 2.5 x 10(-16) mol quantities at injection volumes of 110-120 pL). The detected amounts of fluorescein isothiocyante (FITC)-amino acids are at least ten times lower, since the amino acid:FITC ratio is 10:1 mol. The results demonstrate the perspective of such hybrid PDMS-glass microfluidic systems and the methods to modify their surfaces for on-chip separation methods for biomolecules.     

A simple method for preparation of macroporous polydimethylsiloxane membrane for microfluidic chip-based isoelectric focusing applications

A new, simple method was reported to prepare PDMS membranes with micrometer size pores for microfluidic chip applications. The pores were formed by adding polystyrene and toluene into PDMS prepolymer solution prior to spin-coating and curing. The resulting PDMS membrane has a thickness of around 10 μm and macropores with a diameter ranging from 1 to 2 μm measured using scanning electron microscope (SEM) imaging. This PDMS membrane was validated by integrating it with PDMS microfluidic chips for protein separation using isoelectric focusing mechanism coupled with whole channel imaging detection (IEF-WCID). It has been shown that five standard pI markers and a mixture of two proteins, myoglobin and β-lactoglobulin, can be separated using these chips. The results indicated that this macroporous PDMS membrane can replace the dialysis membrane in PDMS chips for the IEF-WCID technique. The preparation method of macroporous PDMS membrane may be potentially applied in other fields of microfluidic chips.     

聚二甲基硅氧烷基质微流控芯片封接技术的研究

考察了聚二甲基硅氧烷(Polydimethylsiloxane,PDMS)预聚体与固化剂间的配比、固化温度及固化时间对PDMS芯片封接强度的影响,得出PDMS芯片封接的最佳条件基片和盖片所用PDMS预聚体与固化剂质量配比分别为10∶1与5∶1,固化温度为75℃,固化时间分别为35～50min和25～40min,封接后继续加热60min.在该条件下封接制作的微芯片历经半年50多次的分析、冲洗及抽液后未见明显损坏,足以满足一般分析任务的要求,并将芯片成功用于两种氨基酸的快速毛细管电泳分离.     

Electroosmotic flow in a poly(dimethylsiloxane) channel does not depend on percent curing agent

Poly(dimethylsiloxane) (PDMS) microfluidic devices were prepared from different ratios of "curing agent" (which contains silicon hydride groups) to "base" (which contains vinyl-terminated noncross-linked PDMS), to determine the effect of this ratio on electroosmotic flow (EOF). In fabricating devices for this purpose, a novel method for permanently enclosing PDMS channels was developed. As a supplement to the microfluidic method, the inner walls of capillaries were coated with PDMS formed from varying ratios of curing agent to base. EOF was found to be constant for PDMS formed with each ratio, which implies that the negative surface charges do not arise from chemical species present only in the base or the curing agent.     

Microwave plasma treatment of polymer surface for irreversible sealing of microfluidic devices

Microwave plasma was generated in a glass bottle containing 2-3 Torr of oxygen for plasma treatment of a polymer surface. A "kitchen microwave oven" and a dedicated microwave digestion oven were used as the power source. Poly(dimethylsiloxane)(PDMS) slabs treated by a 30 W plasma for 30-60 s sealed irreversibly to form microfluidic devices that can sustain solution flow of an applied pressure of 42 psi without leaking. Experimental set up and conditions for the production of a homogeneous plasma to activate the PDMS surface for irreversible sealing are described in detail. The surface of a microwave plasma-treated PDMS slab was characterized using atomic force microscopy (AFM) and attenuated total reflection-Fourier Transform infrared spectroscopy (ATR-FTIR). The plasma-treated surface bears silica characteristics.