电化学发光检测微流控芯片电学特性分析

本文介绍了电化学发光检测微流控芯片的电学特性,包括电极的阻抗特性,表面磁场特性,电极表面涡流特性以及等效电容分析。电化学以及电化学发光检测通常采用三电极体系,由于加工在微小的尺度上,所以宏观尺度下的电学模型以及电学特性已经不再适用。本文提出了一种新的电化学发光检测微流控芯片电学特性分析的方法,根据微流控芯片与电极的结构与尺寸,建立的微流控芯片电极等效电路模型。不仅可以得出电极间的幅频特性曲线,还可以用于测试外加物理场如电场,磁场或力场和几何参数对芯片电极灵敏度,重复度与线性度的影响。微流控芯片电极等效电路模型与幅频特性分析为研究电化学与电化学发光检测提供了理论依据,为解释微电极的电学特性提供了更好的方法。     

基于激光诱导的细胞荧光成像系统研制

分析了激光诱导荧光法检测钙离子浓度的原理.并利用微流控芯片在细胞培养和检测上的独特优越性,设计实现了基于微流控芯片的测量细胞内钙离子浓度变化的显微荧光成像系统.在对微流控芯片技术研究的基础上设计制作了微流控芯片,并设计了显微系统、快速波长切换系统、CCD成像系统等.利用这套显微荧光成像系统对活体细胞的荧光图像进行采集....     

基于多电容传感法的多电极阵列微流控芯片内 高浓度粒子迁移检测

在浓度密集粒子下对血细胞进行检测与处理可以形成具有特异性的靶向血细胞,对生物和医疗系统的发展具有重要意义。已开发的多电极阵列微流控芯片的电极分布于5个菱形横截面,每个横截面嵌入12个均匀分布的电极。利用该微流控芯片的特殊结构,采用多电容传感法,在大量高浓度粒子中测量了标准聚苯乙烯颗粒的浓度。实验结果表明,在低密度0.3 vol%～1.5 vol%的情况下,粒子迁移率随浓度的增加而成比例地增加。而在中密度1.5 vol%～3.0 vol%的情况下,粒子迁移率增加较少,而在足够密集浓度大于3.0 vol%的情况下,由于颗粒间的相互作用,无论浓度如何变化,粒子迁移率几乎保持恒定。     

无透镜微流控成像流动细胞检测与计数系统

在未来面向个人化的生物医疗诊断中,实时的细胞检测与计数具有重要需求.现有的细胞检测和计数系统例如流式细胞仪和血细胞计数器不适用于小型化流动细胞实时检测和计数.通过将CMOS图像传感器芯片和微流控芯片结合,提出了一种用于流动细胞检测和计数的无透镜微流控成像系统,与用于计数静态细胞的其它无透镜微流控成像系统不同,该系统可以通过基于时域差分的运动检测算法检测和计数微流体通道中连续流动的细胞样本.测试结果表明:该系统可以对微流控通道中流动的人体骨髓基质细胞实现自动检测和计数,并具有-6.53％的低统计错误率.该系统提供了面向未来即时应用的细胞检测和计数解决方案.     

存在故障的数字微流控芯片重构后测试

数字微流控芯片广泛用于生命科学领域,它对可靠性的要求很苛刻。由于数字微流控芯片的可重构性,在测试诊断的故障数小于一定比例时,电极阵列会被重构以撇开故障单元继续使用,而对于重构后的不规则电极单元,必须在使用前做强健完备的测试。首次提出对重构后的不规则电极单元进行并行测试:将重构电极阵列分为多个等大子阵列,每个子阵列分配1个测试液滴进行并行测试,目标为最小化测试时间。本文将测试时间最小化问题转化为分发池的分配问题,并为该NP完全问题建立ILP模型,计算最优测试时间。实验结果表明,该方法避免了重复诊断,最小化了故障后重构芯片的测试时间,获得了较好的测试效果。     

基于FPGA的生物电阻抗成像系统设计

根据电阻抗断层成像技术要求,设计了以Spartan3E系列XC3S500E FPGA为核心的16电极生物电阻抗成像系统,系统嵌入8 bit微处理器PicoBlaze实现逻辑控制并产生激励信号实现高速A/D采集及实现数字解调,通过RS232将采集数据传输到PC机,重建人体内部的电阻率分布或其变化图像。为广泛应用研究电阻抗断层成像技术提供一种硬件设计方案。     

基于柔性印刷工艺的表面肌电电极阵列装置的设计

设计了一种基于柔性印刷工艺的表面肌电电极阵列装置。该电极阵列由12个直径1.2mm的镀金圆电极分成两列组成,内部电极间距为3mm。电极载体材料(聚酰亚胺,厚50μm)具有较高的机械柔性,表面镀金(厚度2μm)的电极具有较低的阻抗,特制的聚酯双面胶带用于可重复使用的电极阵列装置的固定。在单指力量输出任务时记录指浅屈肌的多通道表面肌电(surface Electromyogram,sEMG)信号的实验中得到了稳定的基线和较好的sEMG信号。初步的实验结果表明,设计的这种低成本、体积小的高密度电极阵列装置能用于表面肌肉空间sEMG信号的检测。     

基于图像处理的微流控芯片智能定位系统

针对微流控芯片分析系统中微管道检测手动定位方式定位精度低、耗时费力及无法完成跟踪定位等缺点，设计了一种基于图像处理技术的微流控芯片智能定位系统。系统采用形态学方法、细化算法及Radon变换等相关图像处理方法将芯片平面图上微管道的节点提取出来，生成邻接表，以完成对微流控芯片的智能跟踪定位，并通过定位结果对邻接表进行反馈修正。实验表明所提出的智能跟踪定位方法在对微流控芯片进行跟踪定位时效果良好。     

数字微流控生物芯片布局的拟人遗传组合算法

数字微流控生物芯片布局问题是芯片设计的关键问题,它是在二维微流控阵列上为每个操作布局一个合适的物理位置,以达到完成所有操作的微流控阵列总面积最小和总时间最短两个目标。构建了拟人遗传组合算法,应用拟人启发式算法来控制数字微流控模块的布局过程,用遗传算法对布局结果进行多目标优化,以多元体液检测为实例,模拟了数字微流控生物芯片的布局优化过程。实验结果表明该算法不仅达到了优化目标,且优于并行混合模拟退火算法。     

适用于暗场生化传感系统的微流控芯片的研制与验证

为了提高暗场检测的效率、获取高质量的传感信息,基于实验设备的物理尺寸以及实验环境,通过COMSOL Multiphysics仿真,并采取键合工艺,研制了一种适用于暗场生化传感系统的微流控芯片。该芯片可在同一暗场视野下观察到四个反应区域,实现对多种样品的并行检测。同时由于该设计的连通性,四个反应区域可以两两进行精确对照,对样本检测结果进行定性定量对比。实验中,在暗场显微成像系统下捕获到微流控芯片通入各反应区域的传感单元纳米粒子,测出散射光谱,验证其可行性。暗场显微成像系统和微流控芯片的结合,实现了自动化、集成化的实时原位并行检测,极大地提高了实验效率。     

Thin-film electrode based droplet detection for microfluidic systems

We report on a droplet-producing microfluidic system with electrical impedance-based detection. The microfluidic devices are made of polydimethylsiloxane (PDMS) and glass with thin film electrodes connected to an impedance-monitoring circuit. Immiscible fluids containing the hydrophobic and hydrophilic phases are injected with syringe pumps and spontaneously break into water-in-oil droplet trains. When a droplet passes between a pair of electrodes in a medium having different electrical conductivity, the resulting impedance change signals the presence of the particle for closed-loop feedback during processing. The circuit produces a digital pulse for input into a computer control system. The droplet detector allows estimation of a droplet's arrival time at the microfluidic chip outlet for dispensing applications. Droplet detection is required in applications that count, sort, and direct microfluidic droplets. Because of their low cost and simplicity, microelectrode-based droplet detection techniques should find applications in digital microfluidics and in three-dimensional printing technology for rapid prototyping and biotechnology.     

Human blood microfluidic test chip for imaging,label-free biosensor

We designed and evaluated a microfluidic test chip for human blood filtration and imaging label-free detection of multiple biomarkers. The microfluidic chip has a total size of 75 mm × 25 mm × 2 mm. It is realized as an assembly of a plastic chip and a functionalized photonic crystal slab on a glass substrate. The plastic chip contains capillary channels, a filter membrane and a cavity open on one side. The photonic crystal chip is bonded with an adhesive foil to the open cavity. Human blood filtration is demonstrated. We determined that fluorescently labelled particles of diameter 3 µm or larger are filtered out by the system. Refractometric measurements are performed with the test chip in combination with a compact imaging read-out system to investigate the system response to refractive index changes. Flow dynamics in the sensor cavity are imaged for replacing water by isopropanol. Finally, the binding of 500 nm biotin dissolved in phosphate buffer saline to a photonic crystal surface locally functionalized with streptavidin is demonstrated.     

Development of a biological detection platform utilizing a modular microfluidic stack

The goal of this project is to build a miniaturized, user-friendly cytometry setup (Datta et al. in Microfluidic platform for education and research. COMS, Baton Rouge, 2008; Frische et al. in Development of an miniaturized flow cytometry setup for visual cell inspection and sorting. Baton Rouge, Project Report, 2008) by combining a customized, microfluidic device with visual microscope inspection to detect and extract specific cells from a continuous sample flow. We developed a cytological tool, based on the Coulter particle counter principle, using a microelectrode array patterned on a borosilicate glass chip as electrical detection set-up which is fully embedded into a polymeric multi-layer microfluidic stack. The detection takes place between pairs of coplanar Cr/Au microelectrodes by sensing an impedance change caused by particles continuously carried within a microfluidic channel across the detection area under laminar flow conditions. A wide frequency range available for counting provides information on cell size, membrane capacitance, cytoplasm conductivity and is potentially of interest for in-depth cell diagnostic e.g. to detect damaged or cancerous cells and select them for extraction and further in-depth analysis.     

Method for flow measurement in microfluidic channels based on electrical impedance spectroscopy

We have developed and characterized two novel micro flow sensors based on measuring the electrical impedance of the interface between the flowing liquid and metallic electrodes embedded on the channel walls. These flow sensors are very simple to fabricate and use, are extremely compact and can easily be integrated into most microfluidic systems. One of these devices is a micropore with two tantalum/platinum electrodes on its edges; the other is a micro channel with two tantalum/platinum electrodes placed perpendicular to the channel on its walls. In both sensors the flow rate is measured via the electrical impedance between the two metallic electrodes, which is the impedance of two metal–liquid junctions in series. The dependency of the metal–liquid junction impedance on the flow rate of the liquid has been studied. The effects of different parameters on the sensor’s outputs and its noise behavior are investigated. Design guidelines are extracted and applied to achieve highly sensitive micro flow sensors with low noise.     

Impedance spectra of patch clamp scenarios for single cells immobilized on a lab-on-a-chip

A simple method based on impedance spectroscopy (IS) was developed to distinguish between different patch clamp modes for single cells trapped on microapertures in a patch clamp microchannel array designed for patch clamping on cultured cells. The method allows detecting via impedance analysis whether the cell membrane is ruptured (and culturing prevented) or the cell is still in the attached mode. A modular microfluidic lab-on-a-chip device based on planar patch clamp technology was used to capture multiple individual cells on an array of microapertures. The comparison of the measured and simulated impedance spectra proved that the presented method could distinguish between a cell-attached mode and a whole-cell mode even with low-quality seals. In physiological conditions, the capacitance of HeLa cells was measured to ~38 pF. The first gigaseal was recorded and maintained for 40 min. Once whole-cell configurations were established, trapped cells were superfused with a 140 mM KCl aqueous solution: the change in the measured cell impedance revealed a capacitance decrease to ~27.5 pF that could be due either to a change in the cell size or to the reduced charge separation across the cell membrane. After incubating the chip for 24 h, HeLa cells adhered and grew on the chip surface but did not survive when trapped on the microapertures. The microfluidic system proved to work as a micro electrophysiological analysis system, and the IS-based method can be used for further studies on the post-trapping strength of the seal between the microapertures and the trapped cells to be cultured.     

Thread-based microfluidic system for detection of rapid blood urea nitrogen in whole blood

This study develops a thread-based microfluidic device with variable volume injection capability and 3-dimensional (3D) detection electrodes for capillary electrophoresis electrochemical (CE–EC) detection of blood urea nitrogen (BUN) in whole blood. A poly methyl methacrylate (PMMA) substrate with concave 3D electrodes produced by the hot embossing method is used to enhance the sensing performance of the CE–EC system. Results show that the chip with 3D sensing electrodes exhibits a measured current response nine times higher and signal-to-noise ratio five times higher when compared to the peak responses obtained using a chip with conventional 2D sensing electrodes. In addition, the developed thread-based microfluidic system is capable of injecting variable sample volumes into the separation thread simply by wrapping the injection thread different numbers of times around the separation thread. The peak S/N ratio can be further enhanced with this simple approach. Results also indicate that the CE–EC system exhibits good linear dynamic range for detecting a urea sample in concentrations from 0.1 to 10.0 mM (R ² = 0.9848), which is suitable for adoption in detecting the BUN concentration in human blood (1.78–7.12 mM). Separation and detection of the ammonia ions converted from BUN in whole blood is successfully demonstrated in the present study, with the developed thread-based microfluidic system providing a low-cost, high-performance method for detecting BUN in human blood.     

基于FPGA的高性能三维电阻抗成像系统

构建以FPGA为核心控制器的两层32电极的高性能三维电阻抗成像系统,详细描述了系统的软硬件设计、系统性能测试及成像试验,图像重建采用共轭梯度算法。测试结果表明,系统测量精度达0.082%,系统空间分辨率达0.51%,信噪比达60.3 dB。在盛有盐水的实验盐水槽进行成像试验,结果表明该系统能够准确识别待测区域目标物的个数、位置、大小等信息。系统的构建为深入研究三维电阻抗成像等关键技术提供可靠的硬件平台。     

Electrorotation chip consisting of three-dimensional interdigitated array electrodes

We have developed an electrorotation (ER) chip that has a sandwich structure in which interdigitated array (IDA) electrodes are arranged face-to-face. These IDA electrodes on the top and bottom of the chip were orthogonally arranged to form over 2000 square regions having rotating electric fields between the IDA electrodes. Since rotating electric fields can be generated by arranging the electrical connections to produce a π/2 phase difference between adjacent electrodes, a large number of measurement areas for ER were incorporated within a single ER chip. The ER properties of glass microrods at the individual measurement areas were investigated using this ER chip. The present ER chip was found to be a useful tool for performing high-throughput assays to analyze the dielectric properties of microparticles.