A Label-Free Multisensing Immunosensor Based on Imaging Ellipsometry

An immunosensor based on imaging ellipsometry and its potential applications was demonstrated in this paper. It has been proven a fast, reliable, and convenient method to quantify the thickness distribution of protein layers or detect protein concentration in solution. Combined with a protein chip, the immunosensor was able to detect multiple analytes simultaneously without any labeling. Preliminary results demonstrated how this immunosensor could be used to monitor several independent biospecific binding processes in real-time and in situ conditions.

Feasibility of Protein a for the Oriented Immobilization of Immunoglobulin on Silicon Surface for a Biosensor with Imaging Ellipsometry

The feasibility of using protein A to immobilize antibody on silicon surface for a biosensor with imaging ellipsometry was presented in this study. The amount of human IgG bound with anti-IgG immobilized by the protein A on silicon surface was much more than that bound with anti-IgG immobilized by physical adsorption. The result indicated that the protein A could be used to immobilize antibody molecules in a highly oriented manner and maintain antibody molecular functional configuration on the silicon surface. High reproducibility of the amount of antibody immobilization and homogenous antibody adsorption layer on surfaces could be obtained by this immobilization method. Imaging ellipsometry has been proven to be a fast and reliable detection method and sensitive enough to detect small changes in a molecular monolayer level. The combination of imaging ellipsometry and surface modification with protein A has the potential to be further developed into an efficient immunoassay protein chip.

Preliminary Investigation of Fluid Flow in Meso-Scale Rectangular Tube

The fluid flow associated with micro and meso scale devices is currently of interest. Experiments were performed to study the fluid flow in meso-scale channels. A straight flow tube was fabricated with 1.0x4.0mm^2 in rectangular cross section and 200mm in length, which was made of quartz for flow visualization and PIV measurements. Reynolds numbers were ranged from 311 to over 3105. The corresponding pressure drop was from 0.65KPa to over 16.58KPa between the inlet and outlet of the tube. The micro PIV was developed to measure the velocity distribution in the tube. A set of microscope object lens was mounted ahead of CCD camera to obtain optimized optical magnification on the CCD chip. The velocity distributions near the outlet of the tube were measured to obtain full-developed flow. A CW laser beam was focused directly on the test section by a cylinder lens to form a small light sheet. Thus, high power density of light was formed on the view region. It is very important to the experiment while the velocity of the flow reaches to a few meters per second within millimeter scale. In this case, it is necessary to reduce exposure time to microseconds for PIV measurements. In the present paper, the experimental results are compared with the classical theories.

The Nonlinear Phenomena Of Thin Polydimethylsiloxane (Pdms) Films In Electrowetting

Electrowetting (EW) is an effective way to manipulate small volume liquid in micro- and nano-devices, for it can improve its wettability. Since the late 1990s, electrowetting-on-dielectric (EWOD) has been used widely in bio-MEMS, lab-on-a-chip, etc. Polydimethlsiloxane (PDMS) is extensively utilized as base materials in the fabrication of biomedical micro- and nano-devices. The properties of thin PDMS films used as dielectric layer in EW are studied in this paper. The experimental results show that the thin PDMS films exhibit good properties in EWOD. As to PDMS films with different thicknesses, a threshold voltage and a hysteresis were observed in the EIWOD experiments.

Quantifying Cell Binding Kinetics Mediated By Surface-Bound Blood Type B Antigen To Immobilized Antibodies

Cell adhesion is crucial to many biological processes, such as inflammatory responses, tumor metastasis and thrombosis formation. Recently a commercial surface plasmon resonance (SPR)-based BIAcore biosensor has been extended to determine cell binding mediated by surface-bound biomolecular interactions. How such cell binding is quantitatively governed by kinetic rates and regulating factors, however, has been poorly understood. Here we developed a novel assay to determine the binding kinetics of surface-bound biomolecular interactions using a commercial BIAcore 3000 biosensor. Human red blood cells (RBCs) presenting blood group B antigen and CM5 chip bearing immobilized anti-B monoclonal antibody (mAb) were used to obtain the time courses of response unit, or sensorgrams, when flowing RBCs over the chip surface. A cellular kinetic model was proposed to correlate the sensorgrams with kinetic rates. Impacts of regulating factors, such as cell concentration, flow duration and rate, antibody-presenting level, as well as pH value and osmotic pressure of suspending medium were tested systematically, which imparted the confidence that the approach can be applied to kinetic measurements of cell adhesion mediated by surface-bound biomolecular interactions. These results provided a new insight into quantifying cell binding using a commercial SPR-based BIAcore biosensor.

Role Of Vertical Component Of Surface Tension Of The Droplet On The Elastic Deformation Of Pdms Membrane

Poly(dimethylsiloxane) (PDMS) has been widely used in lab-on-a-chip and micro- total analysis systems (mu-TAS), thus wetting and electrowetting behaviors of PDMS are of great importance in these devices. PDMS is a kind of soft polymer material, so the elastic deformation of PDMS membrane by a droplet cannot be neglected due to the vertical component of the interfacial tension between the liquid and vapor, and this vertical component of liquid-vapor surface tension is also balanced by the stress distribution within the PDMS membrane. Such elastic deformation and stress distribution not only affect the exact measurement of contact angle, but also have influence on the micro-fluidic behavior of the devices. Using ANSYS code, we simulated numerically the elastic deformation and stress distribution of PDMS membrane on a rigid substrate due to the liquid-vapor surface tension. It is found that the vertical elastic deformation of the PDMS membrane is on the order of several tens of nanometers due to the application of a droplet with a diameter of 2.31 mm, which is no longer negligible for lab-on-a-chip and mu-TAS. The vertical elastic deformation increases with the thickness of the PDMS membrane, and there exists a saturated membrane thickness, regarded as a semi-infinite membrane thickness, and the vertical elastic deformation reaches a limiting value when the membrane thickness is equal to or thicker than such saturated thickness. (C) Koninklijke Brill NV, Leiden, 2008.

Measuring Binding Kinetics Of Surface-Bound Molecules Using The Surface Plasmon Resonance Technique

Surface plasmon resonance (SPR) technology and the Biacore biosensor have been widely used to measure the kinetics of biomolecular interactions in the fluid phase. In the past decade, the assay was further extended to measure reaction kinetics when two counterpart molecules are anchored on apposed surfaces. However, the cell binding kinetics has not been well quantified. Here we report development of a cellular kinetic model, combined with experimental procedures for cell binding kinetic measurements, to predict kinetic rates per cell. Human red blood cells coated with bovine serum albumin and anti-BSA monoclonal antibodies (mAbs) immobilized on the chip were used to conduct the measurements. Sensor-grams for BSA-coated RBC binding onto and debinding from the anti-BSA mAb-immobilized chip were obtained using a commercial Biacore 3000 biosensor, and analyzed with the cellular kinetic model developed. Not only did the model fit the data well, but it also predicted cellular on and off-rates as well as binding affinities from curve fitting. The dependence of flow duration, flow rate, and site density of BSA on binding kinetics was tested systematically, which further validated the feasibility and reliability of the new approach. Crown copyright (c) 2008 Published by Elsevier Inc. All rights reserved.

Development Of Biosensor Based On Imaging Ellipsometry

The concept of biosensor based on imaging ellipsometry was proposed ten years ago. Its principle and the methodology as well as some solutions to problems which have to be faced during the development are mentioned. Its properties of phase sensitive, high throughput and fast sampling, as well as label-free, sensitivity better than 1 ng/ml for Immunoglobulin G, and real-time analysis for protein interaction process, etc. provide a potential for applications in biomedicine field. The recent biosensing development with total internal reflection imaging ellipsometry is presented also. [GRAPHICS] An example of 48 protein arrays in matrix. (C) 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

用改进的混合方法分析微电子封装热应变场

介绍一种可用于微电子封装局部应变场分析的实验/计算混合方法，该方法结合了有限元的整体/局部模型和实时的激光云纹干涉技术，利用激光云纹干涉技术所测得的应变场来校核有限元整体模型的计算结果，并用整体模型的结果作为局部模型的边界条件，对实验难以确定的封装结构局部位置的应力、应变场进行分析。用这种方法对可控坍塌倒装封装结构在热载荷作用下焊球内的应变场分布进行了分析，结果表明该方法能够提供封装结构内应力-应变场分布的准确和可靠的结果，为微电子封装的可靠性分析提供重要的依据。

Study of Interaction Force between Antigen and Antibody Using Flow Chamber Method

A parallel plate flow chamber was used to study the interaction force between human IgG (immobilized on a chip surface as ligand) and goat anti-human IgG (immobilized on microspheres surface as receptor). First, it was demonstrated that the binding force between the microspheres and the chip surface came from the bio-specific interaction between the antigen and the antibody. Secondly, it was obtained that the critical shear rate to detach microspheres from the chip surface increases with the ligand surface concentration. Finally, two models to estimate the antigen-antibody bond strength considering bonds' positions were proposed and analyzed.

Formation Mechanism Of Lamellar Chips During Machining Of Bulk Metallic Glass

The uniqThe unique lamellar chips formed in turning–machining of a Vit 1 bulk metallic glass (BMG) are found to be due to repeated shearband formation in the primary shear zone (PSZ). A coupled thermomechanical orthogonal cutting model, taking into account force, free volume and energy balance in the PSZ, is developed to quantitatively characterize lamellar chip formation. Its onset criterion is revealed through a linear perturbation analysis. Lamellar chip formation is understood as a self-sustained limit-cycle phenomenon: there is autonomous feedback in stress, free volume and temperature in the PSZ. The underlying mechanism is the symmetry breaking of free volume flow and source, rather than thermal instability. These results are fundamentally useful for machining BMGs and even for understanding the physical nature of inhomogeneous flow in BMGs.ue lamellar chips formed in turning–machining of a Vit 1 bulk metallic glass (BMG) are found to be due to repeated shearband.

Deformation Of Pdms Membrane And Microcantilever By A Water Droplet: Comparison Between Mooney-Rivlin And Linear Elastic Constitutive Models

In this paper, we studied the role of vertical component Of Surface tension of a water droplet on the deformation of membranes and microcantilevers (MCLs) widely used in lab-on-a-chip and micro-and nano-electromechanical system (MEMS/NEMS). Firstly, a membrane made of a rubber-like material, poly(dimethylsiloxane) (PDMS), was considered. The deformation was investigated using the Mooney-Rivlin (MR) model and the linear elastic constitutive relation, respectively. By comparison between the numerical solutions with two different models, we found that the simple linear elastic model is accurate enough to describe such kind of problem, which would be quite convenient for engineering applications. Furthermore, based on small-deflection beam theory, the effect of a liquid droplet on the deflection of a MCL was also studied. The free-end deflection of the MCL was investigated by considering different cases like a cylindrical droplet, a spherical droplet centered on the MCL and a spherical droplet arbitrarily positioned on the MCL. Numerical simulations demonstrated that the deflection might not be neglected, and showed good agreement with our theoretical analyses. (C) 2008 Elsevier Inc. All rights reserved.

Electrowetting On A Lotus Leaf

Electrowetting on dielectrics has been widely used to manipulate and control microliter or nanoliter liquids in micro-total-analysis systems and laboratory on a chip. We carried out experiments on electrowetting on a lotus leaf, which is quite different from the equipotential plate used in conventional electrowetting. This has not been reported in the past. The lotus leaf is superhydrophobic and a weak conductor, so the droplet can be easily actuated on it through electrical potential gradient. The capillary motion of the droplet was recorded by a high-speed camera. The droplet moved toward the counterelectrode to fulfill the actuation. The actuation speed could be of the order of 10 mm/s. The actuation time is of the order of 10 ms.

Electrowetting on a lotus leaf

Electrowetting on dielectrics has been widely used to manipulate and control microliter or nanoliter liquids in micro-total-analysis systems and laboratory on a chip. We carried out experiments on electrowetting on a lotus leaf which is quite different from the equipotential plate used in conventional electrowetting. This has not been reported in the past. The lotus leaf is superhydrophobic and a weak conductor so the droplet can be easily actuated on it through electrical potential gradient. The capillary motion of the droplet was recorded by a high-speed camera. The droplet moved toward the counterelectrode to fulfill the actuation. The actuation speed could be of the order of 10 mm/s. The actuation time is of the order of 10 ms.

Dual Confocal Laser-Induced Fluorescence/Moveable Contactless Conductivity Detector For Capillary Electrophoresis Microchip

A new dual simultaneous detector was developed for capillary electrophoresis microchip. Confocal laser-induced fluorescence (LIF) and moveable contactless conductivity detection (MCCD) were combined together for the first time. The two detection systems shared a common detection cell and could respond simultaneously. They were mutually independent and advantageous in analyses of mixtures containing organic and inorganic ions. The confocal LIF had high sensitivity and the MCCD could move along the separation channel and detect in different positions of the channel. The detection conditions of the dual detector were optimized. Rhodamine B was used to evaluate the performance of the dual detector. The limit of detection of the confocal LIF was < 5 nM, and that of the MCCD was 0.1 mu M. The dual detector had highly sensitivity and could offer response easily, rapidly and simultaneously.