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.

Biosensor with Total Internal reflection Imaging Ellipsometry

In order to monitor multiple protein reaction processes simultaneously, a biosensor based on imaging ellipsometry operated in the total internal reflection mode is proposed. It could be realised as an automatic analysis for protein interaction processes with real-time label-free method. Its principle and methodology as well as a demonstration for its applications are presented.

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.

Influence of Electrostatic Interaction on Fibrinogen Adsorption on Gold Studied by Imaging Ellipsometry Combined with Electrochemical Methods

Imaging ellipsometry was combined with electrochemical methods for studying electrostatic interactions of protein and solid surfaces. The potential of zero charge for gold-coated silicon wafer/solution interfaces wad determined by AC impedance method. The potential of the gold-coated silicon wafer was controlled at the potential of zero charge, and the adsorption of fibrinogen on the potential-controlled and non-controlled surfaces was measured in real time at the same time by imaging ellipsometry The effect of electrostatic interaction was studied by comparing the difference between the potential of controlled adsorption and the Potential of noncontrolled adsorption. It was shown that the rate of fibrinogen adsorption on the potentiostatic surface was faster than that on the nonpotentiostatic surface. The electrostatic influence on fibrinogen adsorption on the gold-coated silicon wafer was weak, so the hydrophobic interaction should be the major affinity.

Refractive index and thickness analysis of natural silicon dioxide film growing on silicon with variable-angle spectroscopic ellipsometry

The refractive index and thickness of SiO2 thin films naturally grown on Si substrates were determined simultaneously within the wavelength range of 220-1100 nm with variable-angle spectroscopic ellipsometry. Different angles of incidence and wavelength ranges were chosen to enhance the analysis sensitivity for more accurate results. Several optical models describing the practical SiO2-Si system were investigated, and best results were obtained with the optical model, including an interface layer between SiO2 and Si, which proved the existence of the interface layer in this work as described in other publications.

Competitive protein adsorption studied with atomic force microscopy and imaging ellipsometry

The adsorption and competitive adsorption of collagen and bovine serum albumin (BSA) were directly visualized and quantified using atomic force microscopy (AFM) and imaging ellipsometry. Chemically modified silicon surfaces were used as hydrophilic and hydrophobic substrates. The results showed that collagen and BSA in single component solution adsorbed onto a hydrophobic surface two times more than that onto a hydrophilic surface. The competitive adsorption between collagen and BSA showed that serum albumin preferentially adsorbed onto a hydrophobic surface, while collagen on a hydrophilic surface. In the binary solution of BSA (1 mg/ml BSA) and collagen (0.1 mg/ml), nearly 100% of the protein adsorbed onto the hydrophobic surface was BSA, but on the hydrophilic surface only about 6% was BSA. Surface affinity was the main factor controlling the competitive adsorption.

Silicon Surface Modification with a Mixed Silanes Layer to Immobilize Proteins for Biosensor with Imaging Ellipsometry

One kind of surface modification method on silicon wafer was presented in this paper. A mixed silanes layer was used to modify silicon surface and rendered the surface medium hydrophobic. The mixed silanes layer contained two kinds of compounds, aminopropyltriethoxysilane (APTES) and methyltriethoxysilane (NITES). A few of APTES molecules in the layer was used to immobilize covalently human immunoglobulin G (IgG) on the silicon surface. The human IgG molecules immobilized covalently on the modified surface could retain their structures well and bind more antibody molecules than that on silicon surface modified with only APTES. This kind of surface modification method effectively improved the sensitivity of the biosensor with imaging ellipsometry.

Optimization Of Off-Null Ellipsometry For Air/Solid Interfaces

The optimization of off-null ellipsometry is described with emphasis on the improvement of sample thickness sensitivity. Optimal conditions are dependent on azimuth angle settings of the polarizer, compensator, and analyzer in a polarizer-compensator-sample-analyzer ellipsometer arrangement. Numerical simulation utilized offers an approach to present the dependence of the sensitivity on the azimuth angle settings, from which optimal settings corresponding to the best sensitivity are derived. For a series of samples of SiO2 layer (thickness in the range of 1.8-6.5 nm) on silicon substrate, the theory analysis proves that sensitivity at the optimal settings is increased 20 times compared to that at null settings used in most works, and the relationship between intensity and thickness is simplified as a linear type instead of the original nonlinear type, with the relative error reduced to similar to 1/100 at the optimal settings. Furthermore the discussion has been extended toward other factors affecting the sensitivity of the practical system, such as the linear dynamic range of the detector, the signal-to-noise ratio and the intensity from the light source, etc. Experimental results from the investigation Of SiO2 layer on silicon substrate are chosen to verify the optimization. (c) 2007 Optical Society of America.

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.

Optimization of Off-Null Ellipsometry for Optical Biosensor Applications

The optimization of off-null ellipsometry is described with emphasis on the improvement of resolution for visualizing biomolecule layers. For optical biosensor with layer thickness below 6.5 nm, a numerical simulation for the dependence of resolution on the azimuth settings of polarizer and analyzer is presented first. For comparison, three different resolutions are given at three azimuth settings which are near null and far away from null condition, respectively. Furthermore, the square or linear approximation relationship between the intensity and the layer thickness are also given at these settings. The difference among their accuracy is up to 100 times or so. Experimental results of the biosensor sample verify the optimization.

The Development of Biosensor with Imaging Ellipsometry

The concept of biosensor with imaging ellipsometry was proposed about ten years ago. It has become an automatic analysis technique for protein detection with merits of label-free, multi-protein analysis, and real-time analysis for protein interaction process, etc. Its principle, andrelated technique units, such as micro-array, micro-fluidic and bio-molecule interaction cell, sampling unit and calibration for quantitative detection as well as its applications in biomedicine field are presented here.

An auto-focusing method for imaging ellipsometry system

An auto-focusing method based on the image brightness gradient sharpness function is presented for imaging ellipsometry system, in which the image plane of the thin-film specimen is not perpendicular to the optical axis. The clear image of a specimen with large area is obtained by moving the imaging sensor in optical axis direction and around its sensitive surface centre successively. The experimental results demonstrate its feasibility.

Phage M13Ko7 Detection With Biosensor Based On Imaging Ellipsometry And Afm Microscopic Confirmation

A rapid detection and identification of pathogens is important for minimizing transfer and spread of disease. A label-free and multiplex biosensor based on imaging ellipsometry (BIE) had been developed for the detection of phage M13KO7. The surface of silicon wafer is modified with aldehyde, and proteins can be patterned homogeneously and simultaneously on the surface of silicon wafer in an array format by a microfluidic system. Avidin is immobilized on the surface for biotin-anti-M13 immobilization by means of interaction between avidin and biotin, which will serve as ligand against phage M13KO7. Phages M13KO7 are specifically captured by the ligand when phage M13KO7 solution passes over the surface, resulting in a significant increase of mass surface concentration of the anti-M13 binding phage M13KO7 layer, which could be detected by imaging ellipsometry with a sensitivity of 10(9) pfu/ml. Moreover, atomic force microscopy is also used to confirm the fact that phage M13KO7 has been directly captured by ligands on the surface. It indicates that BIE is competent for direct detection of phage M13KO7 and has potential in the field of virus detection. (C) 2008 Elsevier B.V. All rights reserved.

Detection of hepatitis B virus markers using a biosensor based on imaging ellipsometry

A biosensor based on imaging ellipsometry (BIE) has been developed and validated in 169 patients for detecting five markers of hepatitis B virus (HBV) infection. The methodology has been established to pave the way for clinical diagnosis, including ligand screening, determination of the sensitivity, set-up of cut-off values (CoVs) and comparison with other clinical methods. A matrix assay method was established for ligand screening. The CoVs of HBV markers were derived with the help of receiver operating characteristic curves. Enzyme-linked immunosorbent assay (ELISA) was the reference method. Ligands with high bioactivity were selected and sensitivities of 1 ng/mL and 1 IU/mL for hepatitis B surface antigen (HBsAg) and surface antibody (anti-HBs) were obtained respectively. The CoVs of HBsAg, anti-HBs, hepatitis B e antigen, hepatitis B e antibody and core antibody were as follows: 15%, 18%, 15%, 20% and 15%, respectively, which were the percentages over the values of corresponding ligand controls. BIE can simultaneously detect up to five markers within 1 h with results in acceptable agreement with ELISA, and thus shows a potential for diagnosing hepatitis B with high throughput.

Simultaneous phase-shifting ellipsometry based on grating beamsplitter

Simultaneous phase-shifting ellipsometry based on a grating beamsplitter is presented. In the corresponding setup, an orthogonal grating and analyzer array are used. The latter is composed of four separate analyzers arranged in a 2x2 grid, the polarization axes of which are set to 0, 45, 90, and 135 deg. A mask allows only four diffracted beams of the fist order, having the same optical intensities, to pass. Each beam is incident on one of the analyzers of the array. The intensities of the four beams are simultaneously detected by a quadrant detector. The ellipsometric parameters are obtained using the four intensity signals. The feasibility of simultaneous phase shifting ellipsometry is thus demonstrated. (C) 2008 Society of Photo-Optical Instrumentation Engineers.