Novel continuous particle sorting in microfluidic chip utilizing cascaded squeeze effect

This article presents a novel technique for the continuous sorting and collection of microparticles in a microfluidic chip using a cascaded squeeze effect. In the proposed approach, microparticles of different sizes are separated from the sample stream using sheath flows and are then directed to specific side channels for collection. The sheath flows required to separate the particles are generated using a single high voltage supply integrated with a series of variable resistors designed to create electric fields of different intensities at different points of the microchip. Numerical simulations are performed to analyze the electrical potential contours and flow streamlines within the microchannel. Experimental trials show that the microchip is capable of continuously separating microparticles with diameters of 5, 10 and 20 μm, respectively. To further evaluate the performance of the microchip, a sample composed of yeast cells and polystyrene beads is sorted and collected. The results indicate that the microchip achieves a recovery ratio of 87.7% and a yield ratio of 94.1% for the yeast cells and therefore attains a comparable performance to that of a large-scale commercial flow cytometer. Importantly, the high performance of the microchip is achieved without the need for a complex control system or for sophisticated actuation mechanisms such as embedded microelectrodes, ultrasonic generators, or micropumps, and so forth.     

Decreasing microfluidic evaporation loss using the HMDL method: open systems for nucleic acid amplification and analysis

Evaporation is of great importance when dealing with microfluidic devices with open air/liquid interfaces due to the large surface-to-volume ratio. For devices utilizing a thermal reaction (TR) reservoir to perform a series of biological and chemical reactions, excessive heat-induced microfluidic evaporation can quickly lead to reaction reservoir dry out and failure of the overall device. In this study, we present a simple, novel method to decrease heat-induced fluid evaporation within microfluidic systems, which is termed as heat-mediated diffusion-limited (HMDL) method. This method does not need complicated thermal isolation to reduce the interfacial temperature, or external pure water to be added continuously to the TR chamber to compensate for evaporation loss. The principle of the HMDL method is to make use of the evaporated reaction content to increase the vapor concentration in the diffusion channel. The experimental results have shown that the relative evaporation loss (V _loss/V _ini) based on the HMDL method is not only dependent on the HMDL and TR region’s temperatures (T _HMDL and T _TR), but also on the HMDL and TR’s channel geometries. Using the U-shaped uniform channel with a diameter of 200 μm, the V _loss/V _ini within 60 min is low to 5% (T _HMDL = 105°C, T _TR = 95°C). The HMDL method can be used to design open microfluidic systems for nucleic acid amplification and analysis such as isothermal amplification and PCR thermocycling amplification, and a PCR process has been demonstrated by amplifying a 135-bp fragment from Listeria monocytogenes genomic DNA.     

Design,simulation and experiment of electroosmotic microfluidic chip for cell sorting

A microfluidic cell sorting chip has been developed using micromachining technology, where electroosmotic flow (EOF) is exploited to drive and switch cells. For this electroosmotically driven system, it is found that the effect of induced hydrostatic pressure caused by unequal levels in solution reservoirs is not negligible. In this work, the numerical simulation of EOF and opposing pressure induced flow in microchannels is presented and the velocity profiles in the microchannels are measured experimentally using microparticle imaging velocimetry (PIV) system. The result shows that the final resulting velocity is the superposition of the two flows. A total volume of 0.305 μl is transported in the 50 μm microchannel and the back flow occurs after 240 s transportation. The task of sorting cells is realized at the switching structure by adjusting the electric fields in the microchannels. The performance of the cell sorting chip is optimized by investigating the effect of different switching structures. A Y-junction switching structure with 90° switching angle is highly recommended with simulated leakage distance of 53 μm and switching time of 8 ms.     

An integrated silicon sensor with microfluidic chip for monitoring potassium and pH

We present ion-sensitive field effect transistor-based sensors, integrated with a microfluidic chip, for monitoring pH and potassium cations. The sensor is strategically located at the base of a well so that the response time of the device depends both on the mean flow through the device and the diffusion coefficient of the analyte being monitored. This would enable monitoring of ions in the presence of larger molecules. The dependence of the device response time on diffusive transport of analytes was examined through a numerical study of the flow field and the passive diffusion of a chemical species. The predicted device response time was compared with the experimental measurements and reasonable agreement found. The general dependence of device response time on geometry, flow rate, and analyte diffusion coefficient was derived. These devices can be used with biological fluids where monitoring of pH and cations provide vital information about the well-being of patients.     

高聚物微流控芯片上集成化气动微阀的研制

报道了一种新型的聚甲基丙烯酸甲酯(PMMA)/聚二甲基硅氧烷(PDMS)复合芯片。该芯片采用PMMA-PDMS…PDMS-PMMA的四层构型,以在芯片上集成气动微阀。具有液路和控制通道网路的PMMA基片与PDMS弹性膜间采用不可逆封接,分别形成液路半芯片和控制半芯片,而2个半芯片则依靠PDMS膜间的粘性实现可逆封接,组成带有微阀的全芯片。这种制备方法解决了制备PMMA-PDMS-PMMA三层结构芯片的封接难题,封接过程简单可靠。其控制部分和液路部分可以单独更换,可进一步降低使用成本,尤其适合一次性应用场合。初步实验表明:该微阀具有良好的开关性能和耐用性。     

A novel three-dimensional microfluidic platform for on chip multicellular tumor spheroid formation and culture

The formation of three-dimensional (3D) multicellular cell spheroids such as microspheres and embryoid bodies has recently gained much attention as a useful cell culture technique, but few studies have investigated the suitability of glass for spheroids formation and culture. In this work, we present a novel three-dimensional microfluidic device made of poly(dimethylsiloxane) (PDMS) and glass for the easy and rapid synthesis and culture of tumor spheroid. The cell culture unit is composed of an array of microwells on the bottom of a glass plate, bigger microwells and elastomeric microchannels on the top of a PDMS plate. Cell suspension can be easily introduced into the cell culture unit and exchange with the external liquid environment by the microfluidic channels. A single tumor spheroid can be formed and cultured in each glass cell culture chamber, the surface of which was modified with poly(vinyl alcohol) to render it to be resistant to cell adhesion. As the cell culture medium could be replaced, spheroids of the human breast cancer (MCF-7) cells were cultured on the chip for 3 days, reaching the diameters of about 150 μm. Furthermore, the MCF-7 cells were successfully cultured on the chip in 2D and 3D culture modes. Results have shown that glass is well suitable for multicellular tumor spheroids culture. The established platform provides a convenient and rapid method for tumor spheroid culture, which is also adaptable for anticancer drug screening and fundamental biomedical research in cell biology.     

An integrated genetic analysis microfluidic platform with valves and a PCR chip reusability method to avoid contamination

Clinical diagnostics and genomic research often require performing numerous genetic tests. While microfluidic devices provide a low-cost alternative to such demands, integrated microfluidic devices are fabricated using expensive technology not always affordable for single use. However, carryover cross-contamination (CXC) concerns (i.e. either false positive or false negative PCR data) in PCR chips prevent reuse, defying much of the advantages of miniaturized systems developed using expensive MEMS processing. In this work, we present an integrated and reusable PCR–CE glass microfluidic chip capable of multi-chamber PCR and sequential CE, with emphasis on a unique chip reusability approach to avoid CXC. For reliable PCR, the surface of the chamber is re-configured from its virgin hydrophilic (CA < 20°) to hydrophobic (CA > 110°) by silanization. To then extend this silanization method as a chip reusability technique, the silanization coating is ‘stripped and re-silanized’ (SRS) to create a fresh coating prior to each successive PCR run. Experimental confirmation of the effectiveness of SRS method in avoiding the CXC is demonstrated using plasmid DNA and HIV-1 infected DNA samples. We also present passive plug microvalves incorporated in the chip to enable fluid/vapor retention during the PCR and controlled fluid flow from the PCR chamber to the CE section for further analysis.     

An effective PDMS microfluidic chip for chemiluminescence detection of cobalt (II) in water

A microfluidic chip for the chemiluminescence detection of cobalt (II) in water samples, based on the measurement of light emitted from the cobalt (II) catalysed oxidation of luminol by hydrogen peroxide in basic aqueous solution, is presented. The microfluidic chip was designed and fabricated from polydimethylsiloxane using micro-molding method. Optimized reagents conditions were found to be 5.0 × 10^?4 mol/L luminol, 1.0 × 10^?2 mol/L hydrogen peroxide, and 8.0 × 10^?2 mol/L sodium hydroxide. The system can perform fully automated detection with a reagent consumption of only 2.4 μL each time. The linear range of the cobalt (II) ions concentration was 1.0 × 10^?10–1.0 × 10^?3 mol/L and the detection limit was 5.6 × 10^?11 mol/L with the S/N ratio of 3. The relative standard deviation was 4.6 % for 1.0 × 10^?5 mol/L cobalt (II) ions (n = 10).     

Digital microfluidic design and optimization of classic and new fluidic functions for lab on a chip systems

This paper deals with microfluidic studies for lab-on-a-chip development. The first goal was to develop microsystems immediately usable by biologists for complex protocol integrations. All fluid operations are performed on nano-liter droplet independently handled solely by electrowetting on dielectric (EWOD) actuation. A bottom-up architecture was used for chip design due to the development and validation of elementary fluidic designs, which are then assembled. This approach speeds up development and industrialization while minimizing the effort in designing and simplifying chip-fluidic programming. Dispensing reproducibility for 64 nl droplets obtained a CV below 3% and mixing time was only a few seconds. Ease of the integration was demonstrated by performing on chip serial dilutions of 2.8-folds, four times. The second part of this paper concerns the development of new innovative fluidic functions in order to extend EWOD-actuated digital fluidics’ capabilities. Experiments of particle dispensing by EWOD droplet handling are reported. Finally, work is shown concerning the coupling of EWOD actuation and magnetic fields for magnetic bead manipulation.     

A simple 96 well microfluidic chip combined with visual and densitometry detection for resource-poor point of care testing

There is a well-recognized need for low cost biodetection technologies for resource-poor settings with minimal medical infrastructure. Lab-on-a-chip (LOC) technology has the ability to perform biological assays in such settings. The aim of this work is to develop a low cost, high-throughput detection system for the analysis of 96 samples simultaneously outside the laboratory setting. To achieve this aim, several biosensing elements were combined: a syringe operated ELISA lab-on-a-chip (ELISA-LOC) which integrates fluid delivery system into a miniature 96-well plate; a simplified non-enzymatic reporter and detection approach using a gold nanoparticle-antibody conjugate as a secondary antibody and silver enhancement of the visual signal; and carbon nanotubes (CNT) to increase primary antibody immobilization and improve assay sensitivity. Combined, these elements obviate the need for an ELISA washer, electrical power for operation and a sophisticated detector. We demonstrate the use of the device for detection of Staphylococcal enterotoxin B, a major foodborne toxin using three modes of detection, visual detection, CCD camera and document scanner. With visual detection or using a document scanner to measure the signal, the limit of detection (LOD) was 0.5 ng/ml. In addition to visual detection, for precise quantitation of signal using densitometry and a CCD camera, the LOD was 0.1 ng/ml for the CCD analysis and 0.5 ng/ml for the document scanner. The observed sensitivity is in the same range as laboratory-based ELISA testing. The point of care device can analyze 96 samples simultaneously, permitting high throughput diagnostics in the field and in resource poor areas without ready access to laboratory facilities or electricity.     

Development of carbon dot based microplate and microfluidic chip immunoassay for rapid and sensitive detection of HIV-1 p24 antigen

A highly sensitive, precisely specific, environmentally friendly, high-throughput, microwell-plate and microchip-based sandwich assay was developed to detect HIV-1 p24 antigen, a protein biomarker using fluorescent carbon dots. High quantum yield carbon dots were synthesized using citric acid and ethylenediamine as carbon and nitrogen sources by a single-step hydrothermal reaction. The desired amine groups confirmed by FTIR on the carbon dots were coupled to streptavidin by amine–amine coupling reaction using glutaraldehyde. The detection range of the carbon dot based immunoassay (CDIA) was found to be between 20 and 1000 pg/mL in a linear dose-dependent manner. CDIA tested for HIV negative plasma samples showed no false positive results in the detection of HIV-1 p24 antigen. The CDIA was extended to develop a microfluidic carbon dot immunoassay (μCDIA) which exhibited analytical sensitivity in the range of 30–1000 pg/mL. The CDIA and μCDIA can easily be adapted to a lab-on-a-chip platform for use in resource limited settings and can also be multiplexed for the detection of other pathogens like TB and Hepatitis.     

Microfabrication of single-use plastic microfluidic devices for high-throughput screening and DNA analysis

 Modern drug discovery and genomic analysis depend on rapid analysis of large numbers of samples in parallel. The applicability of microfluidic devices in this field needs low cost devices, which can be fabricated in mass production. In close collaboration, Greiner Bio-One and Forschungszentrum Karlsruhe have developed a single-use plastic microfluidic capillary electrophoresis (CE) array in the standardized microplate footprint. Feasibility studies have shown that hot embossing with a mechanical micromachined molding tool is the appropriate technology for low cost mass fabrication. A subsequent sealing of the microchannels allows sub-microliter sample volumes in 96-channel multiplexed microstructures. Received: 16 May 2001 / Accepted: 3 July 2001     

蓝牙BlueCore芯片空间应用可行性试验与分析

提出采用蓝牙BlueCore无线传感器搭建自组织网络进行航天器上的数据无线传输,研究了应用于航天器时影响系统性能和可靠性的关键因素-Bluecore芯片的空间环境工作性能,对其进行了高低温循环和抗电子辐照环境试验。试验结果表明：该芯片可以工作在-70-100℃的温度环境,并可以耐受不低于26krad（Si）的电子辐照,试验数据可以作为航天器的设计和工程应用参考。     

LED芯片检测系统的噪声分析及信号处理研究

分析了LED芯片非接触检测系统噪声信号的功率谱,研究了噪声信号的抑制方法.针对现有检测系统存在检测信噪比低的不足,提出对交变电压信号进行相关检测,采用相关检测后,检测系统的信噪比可以提高20 dB以上.实验结果与仿真结果吻合.该研究对于改进检测系统的性能以满足对多种芯片的正确检测有重要意义.     

Nanomanipulation of single influenza virus using dielectrophoretic concentration and optical tweezers for single virus infection to a specific cell on a microfluidic chip

A major problem when analyzing bionanoparticles such as influenza viruses (approximately 100 nm in size) is the low sample concentrations. We developed a method for manipulating a single virus that employs optical tweezers in conjunction with dielectrophoretic (DEP) concentration of viruses on a microfluidic chip. A polydimethylsiloxane microfluidic chip can be used to stably manipulate a virus. The chip has separate sample and analysis chambers to enable quantitative analysis of the virus functions before and after it has infected a target cell. The DEP force in the sample chamber concentrates the virus and prevents it from adhering to the glass substrate. The concentrated virus is transported to the sample selection section where it is trapped by optical tweezers. The trapped virus is transported to the analysis chamber and it is brought into contact with the target cell to infect it. This paper describes the DEP virus concentration for single virus infection of a specific cell. We concentrated the influenza virus using the DEP force, transported a single virus, and made it contact a specific H292 cell.     

A novel integrated microfluidic platform to perform fluorescence in situ hybridization for chromosomal analysis

The fluorescence in situ hybridization (FISH) technique has been commonly employed to detect the chromosomal abnormalities. However, applications of this technique are limited due to its lengthy process and labor-intensive sample preparation. In this study, a novel integrated microfluidic chip capable of performing the entire FISH protocol automatically was reported. This novel technique can achieve several advantages, including reduce the consumption of bio-samples and reagents, automation and rapid analysis compared to the conventional method. In this study, several functional microfluidic devices were integrated on a single chip to perform automatic FISH on the microfluidic platform. Experimental data demonstrated that the developed microfluidic system successfully provided superior performance for probing the chromosomal abnormality of cells. Furthermore, the novel microfluidic system performed the entire process automatically within 3 h, where the conventional method required 10 h to perform the entire protocol manually. This data indicated superior performance of the novel method. Our findings conclude that the novel integrated FISH protocol is more convenient to perform large quantities of samples, which can be used in clinical trials.     

Structural and microfluidic analysis of hollow side-open polymeric microneedles for transdermal drug delivery applications

In this paper, we present a new design of hollow, out-of-plane polymeric microneedle with cylindrical side-open holes for transdermal drug delivery (TDD) applications. A detailed literature review of existing designs and analysis work on microneedles is first presented to provide a comprehensive reference for researchers working on design and development of micro-electromechanical system (MEMS)-based microneedles and a source for those outside the field who wish to select the best available microneedle design for a specific drug delivery or biomedical application. Then, the performance of the proposed new design of microneedles is numerically characterized in terms of microneedle strength and flow rate at applied inlet pressures. All the previous designs of hollow microneedles have side-open holes in the lumen section with no integrated reservoir on the same chip. We have proposed a new design with side-open holes in the conical section to ensure drug delivery on skin insertion. Furthermore, the present design has an integrated drug reservoir on the back side of the microneedles. Since MEMS-based, hollow, side-open polymeric microneedles with integrated reservoir is a new research area, there is a notable lack of applicable mathematical models to analytically predict structural and fluid flow under various boundary conditions. That is why, finite element (FE) and computational fluid dynamic (CFD) analysis using ANSYS rather than analytical systems has been used to facilitate design optimization before fabrication. The analysis has involved simulation of structural and CFD analysis on three-dimensional model of microneedle array. The effect of axial and transverse loading on the microneedle during skin insertion is investigated in the stress analysis. The analysis predicts that the resultant stresses due to applied bending and axial loads are in the safe range below the yield strength of the material for the proposed design of the microneedles. In CFD analysis, fluid flow rate and pressure drop in the microneedles at applied inlet pressures are numerically and theoretically investigated. The CFD analysis predicts uniform flow through the microneedle array for each microneedle. Theoretical and numerical results for the flow rate and pressure drop are in close agreement with each other, thereby validating the CFD analysis. For the proposed design of microneedles, feasible fabrication techniques such as micro-hot embossing and ultraviolet excimer laser methods are proposed. The results of the present theoretical study provide valuable benchmark and prediction data to fabricate optimized designs of the polymeric, hollow microneedles, which can be successfully integrated with other microfluidic devices for TDD applications.