集成核酸提取的实时荧光PCR微全分析系统

集成核酸提取的实时荧光PCR微全分析系统将核酸提取、PCR扩增与实时荧光检测进行整合,在同一块微流控芯片上实现了核酸分析过程的全自动和全封闭,具有试剂用量少、分析速度快、操作简便等优点。本研究采用微机械加工技术制作集成核酸提取微流控芯片的阳极模,使用组合模具法和注塑法制作具有3D通道的PDMS基片,与玻璃基底通过等离子体键合封装成集成核酸提取芯片。构建了由微流体速度可调节(0~10 mL/min)的驱动控制装置、温控精度可达0.1℃的TEC温控平台、CCD检测功能模块等组成的微全分析系统。以人类血液裂解液为样品,采用硅胶膜进行芯片上核酸提取。系统根据设置好的时序自动执行,以2 mL/min的流体驱动速度完成20μL裂解液上样、清洗;以1 mL/min的流体驱动速度完成DNA洗脱,抽取PCR试剂与之混合注入到反应腔。提取的基因组DNA以链上内参基因GAPDH为检测对象,并以传统手工提取为对照,在该系统平台上进行PCR扩增和熔解曲线分析实验。片上PCR扩增结果显示,扩增曲线明显,Ct值分别为25.3和26.9。扩增产物进行熔解曲线分析得到的熔解温度一致,均为89.9℃。结果表明,此系统能够自动化、全封闭的在微流控芯片上完成核酸提取、PCR扩增与实时定量分析。     

玻璃-PDMS复合芯片微流控气动微阀的研制

报道了一种基于玻璃-PDMS复合芯片微流控气动微阀的制作方法,该方法较Unger等^[1]的方法更为简单易行,并克服了PDMS芯片气动微阀刚性不足和与外流路连接困难的缺点,提高了微阀的可靠性.     

整体式PDMS电泳芯片快速成型及高灵敏化学发光检测氨基酸

通过再铸模法将聚二甲基硅氧烷(PDMS)预聚物固化在由微细金属丝构成的微流体孔道的印模中,一次成型制作了整体式PDMS芯片.将所制作的芯片与化学发光检测器集成构建了微芯片毛细管电泳分析系统.初步考察了不经过衍生化时该系统分离检测氨基酸的性能.实验结果表明,精氨酸和天门冬氨酸在80s内完全分离,分离度为2.45,精氨酸的浓度检测限为3.50μmol/L.     

聚二甲基硅氧烷/玻璃微流控芯片永久性粘合方法及其微通道表面改性研究及其应用

近年来,聚二甲基桂氧烷[Poly(dimethylsilloxane),PDMS]基质微流控芯片因其透光性能好,价格便宜,加工容易,适合大规模生产,成为微全分析系统(Micro total analysis system,μ-TAS)发展的一个热点^[1].PDMS易于复制微通道形状,且具有较高的保真度,省去了玻璃芯片刻蚀的复杂过程;而玻璃具有易于集成功能单元,散热性能好的优点,PDMS-玻璃杂合微流控芯片同时结合了PDMS和玻璃的优点,具有良好的发展前景^[2].     

微流控电阻抗检测系统构建及其应用

凝血时间检测主要用于凝血障碍性疾病的初步诊断、抗凝药物的监测以及外科手术前的评估,是医生对患者自身凝血功能进行评价的一项重要指标。本文应用微流控电阻抗技术,提出了一种基于叉指电极的微流体凝血时间检测方法。使用软刻蚀方式制作微流控芯片,将固化后带有微通道的聚二甲基硅氧烷(PDMS)与带有刻蚀电极的导电玻璃键合,搭建微流控电阻抗检测系统。本实验通过对血浆活化部分凝血活酶时间(APTT)的检测,得出血液凝固过程中阻抗值的变化曲线,引入微积分概念,确定血液凝固时间。     

高聚物芯片毛细管电泳——电化学检测系统的研究

20世纪90年代初发展起来的芯片毛细管电泳技术是现代分析化学领域的发展前沿与研究热点,代表着分析仪器与技术的微型化、自动化、集成化与便携化的发展方向,在生物医学、高通量药物合成与筛选、环境监测和卫生检疫等众多领域有着广阔的应用前景.本论文围绕高聚物毛细管电泳芯片的制备和芯片电泳电化学检测系统的搭建与表征开展了一系列创新性的研究工作:(1)采用光刻与化学刻蚀技术首次制作了不锈钢金属阳模,并详细研究了光刻与蚀刻步骤中的主要工艺参数对不锈钢阳模质量的影响.与传统电铸法制作镍金属阳模相比,本法制作工艺简单、快速,不需要昂贵的仪器设备,制作成本低;与传统的玻璃或硅阳模相比,不锈钢阳模硬度高,寿命长,可多次重复使用.     

微流体芯片动态调控蛋白水溶液pH或Ca~(2+)浓度

模拟蚕和蜘蛛的纺丝系统,以载玻片(75×25mm)为基片,SU-8为阳模材料,聚二甲基硅氧烷(PDMS)为微通道(宽500μm,深100μm)构筑材料,利用光刻及模塑成型等技术设计、制备了微流体芯片,并应用于再生丝素蛋白(RSF)水溶液组成的动态调控。该芯片利用微流体的层流特性使RSF水溶液与pH缓冲液或钙离子缓冲液在微通道内平行流动而不发生混合。在水溶液中RSF质量分数不变的前提下,通过离子扩散实现了流动场下RSF水溶液pH值或钙离子浓度的动态调控。     

一种玻璃微流控芯片的制作方法研究

研究了以ITO膜为掩膜的玻璃微芯片的制作方法和玻璃-玻璃键合技术,并详细讨论了腐蚀条件对掩膜的性能、玻璃的蚀刻速率和微通道表面形貌的影响.总结出了该制作方法与传统玻璃芯片的制作方法相比具有的特点和优势.开发出了一种成本低且简易的玻璃芯片制作方法.     

微泵驱动的微流控芯片流动注射气体扩散分离系统的研究(英文)

本文提出一种毛细-蒸发作用力微泵作为液流驱动力的微流控芯片流动注射气体扩散分离检测系统,该系统可以连续、选择性地检测溶液中的NH+4。微流控芯片为六层结构,包含五层聚二甲基硅氧烷(PDMS)及一层玻璃。采用PDMS加工透气膜,并与其它PDMS层和玻璃层通过等离子体处理后封接。液芯波导管用作检测流通池,以提高检测光程。通过检测溶液中NH+4验证了微流控流动注射系统的性能。系统的检出限(3σ)为40μmol·L-1,分析通量可达60h-1,试样消耗仅100nL。     

声表面波实现微流体垂向输运

提出了声表面波实现微流体垂向输运方法,使得纸基微流器件具有前处理操作功能.在128°旋转Y切割X传播方向的LiNbO3基片上光刻叉指换能器对和反射栅,纸基微流器件通过贴合于压电基片表面的PDMS置放于距压电基片上方2mm处,经功率放大器放大的RF信号加到叉指换能器对上,激发的两相声表面波使得压电基片上待分析微流体垂向运...     

Fabrication of microfluidic systems in poly(dimethylsiloxane)

Microfluidic devices are finding increasing application as analytical systems, biomedical devices, tools for chemistry and biochemistry, and systems for fundamental research. Conventional methods of fabricating microfluidic devices have centered on etching in glass and silicon. Fabrication of microfluidic devices in poly(dimethylsiloxane) (PDMS) by soft lithography provides faster, less expensive routes than these conventional methods to devices that handle aqueous solutions. These soft-lithographic methods are based on rapid prototyping and replica molding and are more accessible to chemists and biologists working under benchtop conditions than are the microelectronics-derived methods because, in soft lithography, devices do not need to be fabricated in a cleanroom. This paper describes devices fabricated in PDMS for separations, patterning of biological and nonbiological material, and components for integrated systems.     

Simple and cheap microfluidic devices for the preparation of monodisperse emulsions

Droplet microfluidics, which can generate monodisperse droplets or bubbles in unlimited numbers, at high speed and with complex structures, have been extensively investigated in chemical and biological fields. However, most current methods for fabricating microfluidic devices, such as glass etching, soft lithography in polydimethylsiloxane (PDMS) or assembly of glass capillaries, are usually either expensive or complicated. Here we report the fabrication of simple and cheap microfluidic devices based on patterned coverslips and microscope glass slides. The advantages of our approach for fabricating microfluidic devices lie in a simple process, inexpensive processing equipment and economical laboratory supplies. The fabricated microfluidic devices feature a flexible design of microchannels, easy spatial patterning of surface wettability, and good chemical compatibility and optical properties. We demonstrate their utilities for generation of monodisperse single and double emulsions with highly controllable flexibility.     

Three-dimensional interconnected microporous poly(dimethylsiloxane) microfluidic devices

This technical note presents a fabrication method and applications of three-dimensional (3D) interconnected microporous poly(dimethylsiloxane) (PDMS) microfluidic devices. Based on soft lithography, the microporous PDMS microfluidic devices were fabricated by molding a mixture of PDMS pre-polymer and sugar particles in a microstructured mold. After curing and demolding, the sugar particles were dissolved and washed away from the microstructured PDMS replica revealing 3D interconnected microporous structures. Other than introducing microporous structures into the PDMS replica, different sizes of sugar particles can be used to alter the surface wettability of the microporous PDMS replica. Oxygen plasma assisted bonding was used to enclose the microstructured microporous PDMS replica using a non-porous PDMS with inlet and outlet holes. A gas absorption reaction using carbon dioxide (CO(2)) gas acidified water was used to demonstrate the advantages and potential applications of the microporous PDMS microfluidic devices. We demonstrated that the acidification rate in the microporous PDMS microfluidic device was approximately 10 times faster than the non-porous PDMS microfluidic device under similar experimental conditions. The microporous PDMS microfluidic devices can also be used in cell culture applications where gas perfusion can improve cell survival and functions.     

A disposable planar peristaltic pump for lab-on-a-chip

We demonstrate a simple planar peristaltic pump fabricated in poly(dimethylsiloxane) (PDMS) via soft lithography and suitable for microfluidic integration.     

Fabrication of circular microfluidic channels by combining mechanical micromilling and soft lithography

The fabrication of microfluidic channels with complex three-dimensional (3D) geometries presents a major challenge to the field of microfluidics, because conventional lithography methods are mainly limited to rectangular cross-sections. In this paper, we demonstrate the use of mechanical micromachining to fabricate microfluidic channels with complex cross-sectional geometries. Micro-scale milling tools are first used to fabricate semi-circular patterns on planar metallic surfaces to create a master mold. The micromilled pattern is then transferred to polydimethylsiloxane (PDMS) through a two-step reverse molding process. Using these semi-circular PDMS channels, circular cross-sectioned microchannels are created by aligning and adhering two channels face-to-face. Straight and serpentine-shaped microchannels were fabricated, and the channel geometry and precision of the metallic master and PDMS molds were assessed through scanning electron microscopy and non-contact profilometry. Channel functionality was tested by perfusion of liquid through the channels. This work demonstrates that micromachining enabled soft lithography is capable of fabricating non-rectangular cross-section channels for microfluidic applications. We believe that this approach will be important for many fields from biomimetics and vascular engineering to microfabrication and microreactor technologies.     

Microfabrication of polydimethylsiloxane electrospray ionization emitters

Microfabricated polydimethylsiloxane (PDMS) emitters for electrospray ionization mass spectrometry (ESI-MS) were implemented as tips along the edge of the PDMS device by three methods which utilize soft lithography processes. These microfabrication methods for producing PDMS emitters as an integral part of a microfluidic device will facilitate development of more complex microfluidic analysis systems using ESI-MS.     

Rapid fabrication of microchannels using microscale plasma activated templating (microPLAT) generated water molds

Poly(dimethylsiloxane) (PDMS) is a common material used in fabricating microfluidic devices. The predominant PDMS fabrication method, soft lithography, relies on photolithography for fabrication of micropatterned molds. In this technical note, we report an alternative molding technique using microscale PLasma Activated Templating (microPLAT). The use of photoresist in soft lithography is replaced by patterned water droplets created using microPLAT. When liquid PDMS encapsulates patterned water and then solidifies, the cavities occupied by water become structures such as microchannels. Using this method, device fabrication is less time consuming, more cost efficient and flexible, and ideal for rapid prototyping. An additional important feature of the water-molding process is that it yields structural profiles that are difficult to achieve using photolithography.     

Components for integrated poly(dimethylsiloxane) microfluidic systems

This review describes the design and fabrication of microfluidic systems in poly(dimethylsiloxane) (PDMS). PDMS is a soft polymer with attractive physical and chemical properties: elasticity, optical transparency, flexible surface chemistry, low permeability to water, and low electrical conductivity. Soft lithography makes fabrication of microfluidic systems in PDMS particularly easy. Integration of components, and interfacing of devices with the user, is also convenient and simpler in PDMS than in systems made in hard materials. Fabrication of both single and multilayer microfluidic systems is straightforward in PDMS. Several components are described in detail: a passive chaotic mixer, pneumatically actuated switches and valves, a magnetic filter, functional membranes, and optical components.     

Integrated electrofluidic circuits: pressure sensing with analog and digital operation functionalities for microfluidics

Microfluidic technology plays an essential role in various lab on a chip devices due to its desired advantages. An automated microfluidic system integrated with actuators and sensors can further achieve better controllability. A number of microfluidic actuation schemes have been well developed. In contrast, most of the existing sensing methods still heavily rely on optical observations and external transducers, which have drawbacks including: costly instrumentation, professional operation, tedious interfacing, and difficulties of scaling up and further signal processing. This paper reports the concept of electrofluidic circuits - electrical circuits which are constructed using ionic liquid (IL)-filled fluidic channels. The developed electrofluidic circuits can be fabricated using a well-developed multi-layer soft lithography (MSL) process with polydimethylsiloxane (PDMS) microfluidic channels. Electrofluidic circuits allow seamless integration of pressure sensors with analog and digital operation functions into microfluidic systems and provide electrical readouts for further signal processing. In the experiments, the analog operation device is constructed based on electrofluidic Wheatstone bridge circuits with electrical outputs of the addition and subtraction results of the applied pressures. The digital operation (AND, OR, and XOR) devices are constructed using the electrofluidic pressure controlled switches, and output electrical signals of digital operations of the applied pressures. The experimental results demonstrate the designed functions for analog and digital operations of applied pressures are successfully achieved using the developed electrofluidic circuits, making them promising to develop integrated microfluidic systems with capabilities of precise pressure monitoring and further feedback control for advanced lab on a chip applications.     

Design and fabrication of chemically robust three-dimensional microfluidic valves

A current problem in microfluidics is that poly(dimethylsiloxane) (PDMS), used to fabricate many microfluidic devices, is not compatible with most organic solvents. Fluorinated compounds are more chemically robust than PDMS but, historically, it has been nearly impossible to construct valves out of them by multilayer soft lithography (MSL) due to the difficulty of bonding layers made of "non-stick" fluoropolymers necessary to create traditional microfluidic valves. With our new three-dimensional (3D) valve design we can fabricate microfluidic devices from fluorinated compounds in a single monolithic layer that is resistant to most organic solvents with minimal swelling. This paper describes the design and development of 3D microfluidic valves by molding of a perfluoropolyether, termed Sifel, onto printed wax molds. The fabrication of Sifel-based microfluidic devices using this technique has great potential in chemical synthesis and analysis.