Passive microfluidic devices for plasma extraction from whole human blood

Our goal is to analyze and compare different continuous microfluidic principles dedicated to plasma extraction from hardly diluted human blood for lab-on-chip applications. First, the strengths and weaknesses of various emerging passive microfluidic methods (microfiltration- and centrifugation-based methods) were analyzed. Various devices were designed, microfabricated and tested with beads or blood. Filtration may be efficient, but with a high sample dilution, low flow rate and optimized geometry. Due to fast cell clogging, this remains a short-term solution. Separation effects resulting from centrifugal acceleration in curved channel flows are hindered by Dean vortices and anyhow are not pronounced with blood. An innovative device is then proposed and investigated experimentally. This is based on the lateral migration of red cells and the resulting cell-free layer, which is used to supply geometric singularities (an ear-cavity or a corner-edge) and locally enhance the clear plasma region. A maximum extraction of 10.7% is obtained for 1/20 diluted blood, injected at 100 μL/min in the corner-edge design.     

An automatic microfluidic system for rapid screening of cancer stem-like cell-specific aptamers

This study reports a microfluidic system which automatically performs the systematic evolution of ligands by exponential enrichment (SELEX) process for rapid screening of aptamers which are specific to cancer stem-like cells. The system utilizes magnetic bead-based techniques to select DNA aptamers and has several advantages including a rapid, automated screening process, and less consumption of cells and reagents. By integrating a microfluidic control module, a magnetic bead-based aptamer extraction module, and a temperature control module, the entire Cell-SELEX process can be performed in a shorter period of time. Compared with the traditional Cell-SELEX process, this microfluidic system is more efficient and consumes fewer sample volumes. It only takes approximately 3 days for an entire Cell-SELEX process with 15 screening runs, which is relatively faster than that of a traditional Cell-SELEX process (1 week for 15 rounds). The binding affinity of this resulting specific aptamer was measured by a flow cytometric analysis to have a dissociation constant (K _d) of 15.32 nM. The capture rate for cancer stem-like cells using the specific aptamer-conjugated bead is better than that using Ber-EP4 antibody-conjugated bead. This microfluidic system may provide a powerful platform for the rapid screening of cell-specific aptamers.     

An integrated microfluidic system for fast, automatic detection of C-reactive protein

C-reactive protein (CRP) is a well-known inflammation marker in human beings. This study reports a new microfluidic system for fast, automatic detection of CRP. It contains pneumatic micropumps, a vortex-type micromixer, a pneumatic micro-injector and several microvalves to automatically perform the entire protocol for CRP detection. This includes sample/reagent transportation, incubation between the target CRP and a CRP-specific aptamer, washing processes, and the chemiluminescence development process. In addition, the chemiluminescence signal is measured by using a custom-made optical system which consists of a photomultiplier tube, a portable air compressor and eight electronic magnet valves to quantify the concentration of CRP. When compared to previous works, not only can this new microfluidic system automatically perform the entire process via a new integrated micro-injector and new micropumps, but a new CRP-specific DNA aptamer with a higher affinity and specificity is also used for CRP measurement. Experimental data show that the developed system can automatically complete the entire protocol within 30 min with a detection limit of 0.0125 mg/L, which is superior to previous published results. Moreover, this study also measures CRP concentration from clinical samples to verify the performance of the developed microfluidic system. The results indicate that the measured CRP concentrations from human serums are consistent with those using a benchtop system. The developed system can also detect CRP concentrations from human whole blood without any external sample pretreatment process. This microfluidic system may be promising for point-of-care applications for CRP detection in the future.     

水体氨氮原位快速检测智能传感器的研制

介绍了氨氮传感器的检测原理,给出了智能氨氮传感器的结构设计和电路设计方法,重点讲述了氨气敏和铵离子传感器的标定算法以及多传感器数据融合算法,并对其性能进行了测试和分析。该智能氨氮传感器是集成了氨气敏、铵离子和pH温度探头的复合传感器,可以用于实现水体氨氮含量的原位快速检测。     

A bead-based microfluidic system for joint detection in TORCH screening at point-of-care testing

Microsystem Technologies - A concise bead-based microfluidic system has been developed for joint detection in TORCH screening at point-of-care testing. Assisted by five functionalized polycarbonate...     

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.     

Capillary flow-driven blood plasma separation and on-chip analyte detection in microfluidic devices

We report a comprehensive review on the capillary flow-driven blood plasma separation and on-chip analyte detection in microfluidic devices. Blood plasma separation is the primary sample preparation step prior to most biochemical assays. Conventionally, centrifugation is used for the sample preparation process. There are numerous works reporting blood plasma separation in microfluidic devices which aim at miniaturizing the sample preparation procedure. Capillary-based blood plasma separation shows promise in actualizing point-of-care diagnostic devices for applications in resource-limited settings including military camps and rural areas. In this review, the devices have been categorized based on active and passive plasma separation techniques used for the separation of plasma from capillary-driven blood sample. A comparison between different techniques used for blood plasma separation is outlined. On-chip detection of analytes present in the separated plasma obtained using some of these reported devices is also presented and discussed.     

纸尿裤标识快速视觉检测系统研究

为解决纸尿裤生产线上人工检测效率低下的问题,利用Visual C++6.0开发了尿裤标识视觉检测系统.采用彩色工业相机,实时抓取产品图像,对纸尿裤弹性腰围位置和左右腰帖的角度做出分析和判断.弹性腰围检测通过对图像进行亮度、对比度调整,HSI空间颜色提取,动态二值化等方法将弹性腰围从背景中分离出来,通过统计目标区域中有效点的个数判定腰围的位置.腰帖检测采用动态二值化和边缘检测相结合的方法,提取腰帖和背景的边界线,通过扫描法获得腰帖角度.实验证明:该系统可以有效解决光照不足、低对比度、褶皱干扰等问题,稳定地实现纸尿裤标识的在线检测.     

Multi-step microfluidic system for blood plasma separation: architecture and separation efficiency

This document presents a multi-step microfluidic system designed for passive and continuous separation of human blood plasma. This system is based on lateral migration of red blood cells, which leads to a cell-free layer close to the wall. The geometry of the plasma separation unit used in this system was determined by research conducted by Sollier et al. in Biomed Microdevices 12:485–497 (2010). It makes the most of geometric singularities to increase the size of the cell-free layer and optimise the quantity of plasma recovered. This article endeavours to show the importance of architectural fluidic connection upstream of the cell on plasma yield. The design of the chip was then modified to remove its connection role. It was then possible to consider installing the yield cell in series. The approach used for the overall optimisation of the system is presented in the article. In the case of two successive patterns, the increase in pure (diluted) plasma yield ranges from 18 to 25 % for 1:20 diluted blood, and the quality of the plasma obtained is compared to traditional separation methods.     

A paper-based microfluidic Dot-ELISA system with smartphone for the detection of influenza A

An automated, portable, and integrated paper-based microfluidic system has been developed for influenza A detection with smartphone at point-of-care (POC) settings. The low-cost paper-based microfluidic chip consists of a reagent storage and reaction modules. The storage module, which consists of a couple of reagent chambers with dispensation channels, is responsible for reagent storage and release. The reaction module consists of an absorbent pad and a nitrocellulose (NC) membrane which is functionalized with specific monoclonal antibodies on a test and control spots for immunoassay detection. Microfluidic Dot-ELISA is performed when the dispensed reagent flows through the NC membrane at a controllable speed and reaches the absorbent pad because of the gravity and capillary force without active pumping. A smartphone is used to capture image from the NC membrane with its own camera and process the image with an intelligent algorithm of custom application software which is developed with Java. With a smartphone, the detection result can be displayed and transmitted to other medical agencies if necessary. Experimental results show that, compared with the traditional methods, more convenient and efficient influenza A detection can be achieved with the developed paper-based POC microfluidic chip with the assistance of smartphone.     

Utilization of nanoparticles in microfluidic systems for optical detection

An increasing interest has been shown in microfluidic systems due to their properties including low consumption of reagents, short analysis time and easy integration. However, despite of these advantages over conventional methods, some limitations in sensitivity and selectivity still exist in microfluidic systems. Recently advancements in nanotechnology offer some new approaches for the detection of target analytes with high sensitivity and selectivity. As a result, it is an appropriate method to enhance the detection sensitivity through a combination between microfluidic system and nanotechnology. Optical detection is a dominant technique in microfluidics because of its noninvasive nature and easy coupling. Numerous studies that integrate optical microfluidic system with nanotechnology have been reported in recent years. Therefore, optical microfluidic systems in combination with nanomaterials (NMs) are reviewed in our work. We illustrate the functions of different NMs in optical microfluidic systems and the efforts of different researchers to improve the performance of devices. After the introduction of different nanoparticle-based optical detection methods, challenges and future directions in the development of nanoparticle-based optical detection schemes in microfluidics have also been discussed.     

A microfluidic device for thermal particle detection

We demonstrate the use of heat to count microscopic particles. A thermal particle detector (TPD) was fabricated by combining a 500-nm-thick silicon nitride membrane containing a thin-film resistive temperature detector with a silicone elastomer microchannel. Particles with diameters of 90 and 200 μm created relative temperature changes of 0.11 and ?0.44 K, respectively, as they flowed by the sensor. A first-order lumped thermal model was developed to predict the temperature changes. Multiple particles were counted in series to demonstrate the utility of the TPD as a particle counter.     

A disposable single-use electrochemical sensor for the detection of uric acid in human whole blood

We report here a non-enzymatic detecting electrode strip for fast monitoring of uric acid in human whole blood. A single-use amperometric uric acid sensor strip, incorporating a three-electrode configuration, has been fabricated on a polypropylene substrate using low cost screen-printing (thick-film) technology. Both the working and counter electrodes were prepared by screen-printing commercial carbon ink. The integration on the same support of pseudo-reference electrode was obtained by screen-printing a commercial silver ink and subsequent electrochemical pretreatment. Simply by placing a 20 μl human whole blood drop on the strip is enough for uric acid analysis by square-wave voltammetry. Real human whole blood samples were analyzed by this method and compared to the phosphotungstic acid clinical test procedure with satisfactory results.     

Positioning system for particles in microfluidic structures

Fast continuous flow detection of biomolecules in lab-on-a-chip structures is a challenging task. Combining these molecules with small magnetic particles, the interaction between their stray field and, e.g., magneto-resistive sensors can be used to indirectly prove the biomolecules. To position the particles on top of a sensor array at the bottom of the flow channel, we propose a microfluidic structure of changing channel height combining hydrodynamic and gravitational effects. We present numerical calculations predicting an increase in the capture rate by more than 100% in comparison to a straight channel. We experimentally realize an optical analysis of the specific binding of biotin-functionalized Chemagen beads on a streptavidin-coated surface. To prove the binding is not due to the surface effects, a second uncoated bead species is employed.     

用于血样前处理的BioMEMS微流控芯片

基于BioMEMS技术,研制成三种血液样品前处理微流控芯片,分别介绍了血样前处理微流控芯片的原理、结构、制备技术以及样品前处理效果.基于错流过滤原理,设计了用于血细胞分离的错流过滤微结构,采用深刻蚀技术在硅片上刻蚀出直径为20μm,高度为50 μm的圆柱阵列;基于化学法破裂细胞,设计了用于血细胞破裂的夹流式微沟道,采用湿法腐蚀技术在硅片上腐蚀出深度约为80μm的微沟道;基于固相萃取原理,设计了用于DNA提纯的介孔固相载体,采用电化学阳极腐蚀技术在硅微沟道内表面制得表面积为300m2/g的介孔层.分别在芯片上实现了血细胞的分离、血细胞的破裂以及DNA的提纯.     

一种用于有机磷农药检测的微流控传感器

成功制作出一种用于检测有机磷农药的聚二甲基硅氧烷(PDMS)微流控传感器,该传感器集成了光纤和用于固定有机磷水解酶的SU-8圆柱.检测原理基于反应产物对光的吸收,不同浓度有机磷农药的实验结果显示:该传感器的线性相关系数为0.9934.实验结果展示了一种将酶固定在微流沟道的简单方法.简单的制备工艺和便携式的器件为集成化的...     

An integrated microfluidic system using mannose-binding lectin for bacteria isolation and biofilm-related gene detection

Molecular diagnosis of biofilm-related genes (BRGs) in common bacteria that cause periprosthetic joint infections may provide crucial information for clinicians. In this study, several BRGs, including ica, fnbA, and fnbB, were rapidly detected (within 1 h) with a new integrated microfluidic system. Mannose-binding lectin (MBL)-coated magnetic beads were used to isolate these bacteria, and on-chip nucleic acid amplification (polymerase chain reaction, PCR) was then performed to detect BRGs. Both eukaryotic and prokaryotic MBLs were able to isolate common bacterial strains, regardless of their antibiotic resistance, and limits of detection were as low as 3 and 9 CFU for methicillin-resistant Staphylococcus aureus and Escherichia coli, respectively, when using a universal 16S rRNA PCR assay for bacterial identification. It is worth noting that the entire process including bacteria isolation by using MBL-coated beads for sample pre-treatment, on-chip PCR, and fluorescent signal detection could be completed on an integrated microfluidic system within 1 h. This is the first time that an integrated microfluidic system capable of detecting BRGs by using MBL as a universal capturing probe was reported. This integrated microfluidic system might therefore prove useful for monitoring profiles of BRGs and give clinicians more clues for their clinical judgments in the near future.     

Thread-based probabilistic models for expert finding in enterprise Microblogs

Within the enterprise important conversations are shifting to Social Media Microblogs thus providing a new source for the acquisition of tacit knowledge useful for intelligent agents. While automated retrieval of candidate experts from Web documents has been well-researched, little exists that leverages social aspects of Microblogs for this purpose. We show that the experts and their related expertise can also be identified in the enterprise corpus. Analysis reveals the existence of problem-solving threads where the requestor seeking help often elicits responses from experts which consequently records tacit (experiential) knowledge. Here we present rules to automate the acquisition of this tacit knowledge. The rules are based on probabilistic models enhanced with linguistics to exploit the role patterns in threads. Heuristics also strengthen local evidence about associations between candidates, documents and (expertise) terms. Applying this, we demonstrate that the Enhanced Models can ameliorate the negative impact of Microblogs (such as sparse data, short content, and implicit associations). The experiments show that the candidate models significantly outperform the document models in the Microblog environment. This is different from previous research in Web environments. Examples also illustrate underlying insights and emphasize the ‘additive’ nature of expertise found in Social Media.