Determination of SARS-coronavirus by a microfluidic chip system

We have developed a new experimental system based on a microfluidic chip to determine severe acute respiratory syndrome coronavirus (SARS-Cov). The system includes a laser-induced fluorescence microfluidic chip analyzer, a glass microchip for both polymerase chain reaction (PCR) and capillary electrophoresis, a chip thermal cycler based on dual Peltier thermoelectric elements, a reverse transcription-polymerase chain reaction (RT-PCR) SARS diagnostic kit, and a DNA electrophoretic sizing kit. The system allows efficient cDNA amplification of SARS-CoV followed by electrophoretic sizing and detection on the same chip. To enhance the reliability of RT-PCR on SARS-CoV detection, duplex PCR was developed on the microchip. The assay was carried out on a home-made microfluidic chip system. The positive and the negative control were cDNA fragments of SARS-CoV and parainfluenza virus, respectively. The test results showed that 17 positive samples were obtained among 18 samples of nasopharyngeal swabs from clinically diagnosed SARS patients. However, 12 positive results from the same 18 samples were obtained by the conventional RT-PCR with agarose gel electrophoresis detection. The SARS virus species can be analyzed with high positive rate and rapidity on the microfluidic chip system.

Simultaneous Laser-Induced Fluorescence And Contactless-Conductivity Detection For Microfluidic Chip

A combined detection system involving simultaneous LIF and contactless-conductometric measurements at the same place of the microfluidic chip was described. The LIF measurement was designed according to the confocal principle and a moveable contactless-conductivity detector was used in (CD)-D-4. Both measurements were mutually independent and advantageous in analyses of mixtures. Various experimental parameters affecting the response were examined and optimized. The performances were demonstrated by simultaneous detection of Rhodamine B. And the results showed that the combined detection system could be used sensitively and reliably. (C) 2008 Yong Yu. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Developing A Microfluidic-Based System To Quantify Cell Capture Efficiency

Micro-fabrication technology has substantial potential for identifying molecular markers expressed on the surfaces of tissue cells and viruses. It has been found in several conceptual prototypes that cells with such markers are able to be captured by their antibodies immobilized on microchannel substrates and unbound cells are flushed out by a driven flow. The feasibility and reliability of such a microfluidic-based assay, however, remains to be further tested. In the current work, we developed a microfluidic-based system consisting of a microfluidic chip, an image grabbing unit, data acquisition and analysis software, as well as a supporting base. Specific binding of CD59-expressed or BSA-coupled human red blood cells (RBCs) to anti-CD59 or anti-BSA antibody-immobilized chip surfaces was quantified by capture efficiency and by the fraction of bound cells. Impacts of respective flow rate, cell concentration, antibody concentration and site density were tested systematically. The measured data indicated that the assay was robust. The robustness was further confirmed by capture efficiencies measured from an independent ELISA-based cell binding assay. These results demonstrated that the system developed provided a new platform to effectively quantify cellular surface markers effectively, which promoted the potential applications in both biological studies and clinical diagnoses.

Integrated lectin affinity microfluidic chip for glycoform separation

Lectin affinity chromatography was miniaturized into a microfluidic format, which results in improvement of performance, as compared to the conventional method. A lectin affinity monolith column was prepared in the microchannel of a microfluidic chip. The porous monolith was fabricated by UV-initiated polymerization of ethylene dimethacrylate (EDMA) and glycidyl methacrylate (GMA) in the presence of porogeneities, followed by immobilization of pisum sativum agglutinin (PSA) on the monolith matrix. Using electroosmosis as the driven force, lectin affinity chromatographies of three kinds of glycoprotein, turkey ovalbumin (TO), chicken ovalbumin (CO), and ovomucoid (OM), were carried out on the microfluidic system. All the glycoproteins were successfully separated into several fractions with different affinities toward the immobilized PSA. The integrated system reduces the time required for the lectin affinity chromatography reaction to similar to3%, thus, the overall analysis time from 4 h to 400 s. Only 300 pg of glycoprotein is required for the whole separation process. Moreover, troublesome operations for lectin affinity chromatography are simplified.

Injection by hydrostatic pressure in conjunction with electrokinetic force on a microfluidic chip

A simple method was developed for injecting a sample on a cross-form microfluidic chip by means of hydrostatic pressure combined with electrokinetic forces. The hydrostatic pressure was generated simply by adjusting the liquid level in different reservoirs without any additional driven equipment such as a pump. Two dispensing strategies using a floating injection and a gated injection, coupled with hydrostatic pressure loading, were tested. The fluorescence observation verified the feasibility of hydrostatic pressure loading in the separation of a mixture of fluorescein sodium salt and fluorescein isothiocyanate. This method was proved to be effective in leading cells to a separation channel for single cell analysis.

面向微粒操纵的介电泳芯片系统研究

针对目前在纳米器件及传感器的制造中尚无对大量粒子进行有效操纵的方法,我们利用介电泳方法对大量微粒进行定位和传输操纵,介绍了利用MEMS工艺进行介电泳芯片加工的过程以及整个观测与实验系统的建立,通过有限元软件对传统介电泳和行波介电泳中电极阵列的电场分布进行求解,并在该实验系统下实现了对微通道中的悬浮高度和微粒的运动速度的测量.该实验系统的研究为液体环境下微纳颗粒的装配和分离提供了一条有效的技术路径.

A Compact Direct Digital Frequency Synthesizer for System-on-chip

A compact direct digital frequency synthesizer (DDFS) for system-on-chip (SoC) is developed in this paper. For smaller chip size and lower power consumption, the phase to sine mapping data is compressed by using sine symmetry technique, sine-phase difference technique, quad line approximation (QLA) technique and quantization and error read only memory (QE-ROM) technique. The ROM size is reduced by 98 % using the techniques mentioned above. A compact DDFS chip with 32-bit phase storage depth and a 10-bit on-chip digital to analog converter(DAC) has been successfully implemented using standard 0.35um CMOS process. The core area of the DDFS is 1.6mm(2). It consumes 167 mW at 3.3V, and its spurious free dynamic range (SFDR) is 61dB.

A Label-Free Protein Microfluidic Array for Parallel Immunoassays

A label-free protein microfluidic array for immunoassays based on the combination of imaging ellipsometry and an integrated microfluidic system is presented. Proteins can be patterned homogeneously on substrate in array format by the microfluidic system simultaneously. After preparation, the protein array can be packed in the microfluidic system which is full of buffer so that proteins are not exposed to denaturing conditions. With simple microfluidic channel junction, the protein microfluidic array can be used in serial or parallel format to analyze single or multiple samples simultaneously. Imaging ellipsometry is used for the protein array reading with a label-free format. The biological and medical applications of the label-free protein microfluidic array are demonstrated by screening for antibody–antigen interactions, measuring the concentration of the protein solution and detecting five markers of hepatitis B.

Microfluidic effects of transporting signaling components in cell coculture chips

A theoretical model has been developed to investigate the microfluidic transport of the signaling chemicals in the cell coculture chips. Using an epidermal growth factor (EGF)-like growth factor as the sample chemical, the effects of velocities and channel geometry were studied for the continuous-flow microchannel bioreactors. It is found that different perfusion velocities must be applied in the parallel channels to facilitate the communication, i.e., transport of the signaling component, between the coculture channels. Such communication occurs in a unidirectional way because the signaling chemicals can only flow from the high velocity area to the low velocity area. Moreover, the effect of the transport of the signaling component between the coculture channels on the growth of the monolayer cells and the multicellular tumor spheroid (MTS) in the continuous-flow coculture environment were simulated using 3D models. The numerical results demonstrated that the concentration gradients will induce the heterogeneous growth of the cells and the MTSs, which should be taken into account in designing the continuous-flow perfusion bioreactor for the cell coculture research.

Identification and characterization of Bacillus anthracis by multiplex PCR on DNA chip

Bacillus anthracis can be identified by detecting virulence factor genes located on two plasmids, pXO1 and pXO2. Combining multiplex PCR with arrayed anchored primer PCR and biotin-avidin alkaline phosphatase indicator system, we developed a qualitative DNA chip method for characterization of B. anthracis, and simultaneous confirmation of the species identity independent of plasmid contents. The assay amplifies pag gene (in pXO1), cap gene (in pXO2) and Ba813 gene (a B. anthracis specific chromosomal marker), and the results were indicated by an easy-to-read profile based on the color reaction of alkaline phosphatase. About 1 pg of specific DNA fragments on the chip wells could be detected after PCR. With the proposed method, the avirulent (pXO1(+)/2(-), pXO1(-)/2(+) and pXO1(-)/2(-)) strains of B. anthracis and distinguished 'anthrax-like' strains from other B. cereus group bacteria were unambiguously identified, while the genera other than Bacillus gave no positive signal. (C) 2004 Elsevier B.V. All rights reserved.

A Complex BPF with On Chip Auto-tuning Architecture for Wireless Receivers

A 3(rd) order complex band-pass filter (BPF) with auto-tuning architecture is proposed in this paper. It is implemented in 0.18um standard CMOS technology. The complex filter is centered at 4.092MHz with bandwidth of 2.4MHz. The in-band 3(rd) order harmonic input intercept point (IIP3) is larger than 16.2dBm, with 50 Omega as the source impedance. The input referred noise is about 80uV(rms). The RC tuning is based on Binary Search Algorithm (BSA) with tuning accuracy of 3%. The chip area of the tuning system is 0.28 x 0.22 mm(2), less than 1/8 of that of the main-filter which is 0.92 x 0.59 mm(2). After tuning is completed, the tuning system will be turned off automatically to save power and to avoid interference. The complex filter consumes 2.6mA with a 1.8V power supply.

A Complex BPF with On Chip Auto-tuning Architecture for Low-IF Receivers

A 3(rd) order complex band-pass filter (BPF) with auto-tuning architecture is proposed in this paper. It is implemented in 0.18 mu m standard CMOS technology. The complex filter is centered at 4.092MHz with bandwidth of 2.4MHz. The in-band 3(rd) order harmonic input intercept point (IIP3) is larger than 19dBm, with 50 Omega as the source impedance. The input referred noise is about 80 mu V-rms. The RC tuning is based on Binary Search Algorithm (BSA) with tuning accuracy of 3%. The chip area of the tuning system is 0.28x0.22mm(2), less than 1/8 of that of the main-filter which is 0.92x0.59mm(2). After tuning is completed, the tuning system will be turned off automatically to save power and to avoid interference. The complex filter consumes 2.6mA with a 1.8V power supply.

A low power integrated CMOS transmitter for BCI system

The prototype wafer of a low power integrated CMOS Transmitter for short-range biotelemetry application has been designed and fabricated, which is prospective to be implanted in the human brain to transfer the extracted neural information to the external computer. The transmitter consists of five parts, a bandgap current regulator, a ring oscillator, a buffer, a modulator and a power transistor. High integration and low power are the most distinct criteria for such an implantable integrated circuit. The post-simulation results show that under a 3.3 V power supply the transmitter provides 100.1 MHz half-wave sinusoid current signal to drive the off-chip antenna, the output peak current range is -0.155 mA similar to 1.250 mA, and on-chip static power dissipation is low to 0.374 mW. All the performances of the transmitter satisfy the demands of wireless real-time BCI system for neural signals recording and processing.