Study of a Paraboloidal Lens-based Surface Plasmon Resonance Instrument for Bioaffinity Interactions of Molecules in a Biochip
Date Issued
2004
Date
2004
Author(s)
Lee, Shu-Sheng
DOI
zh-TW
Abstract
The instrument design for surface plasmon resonance (SPR) coupled heavily with the principle of surface plasmon waves. When a light beam impinges onto the interface between the dielectric media and metal media at a critical angle, a surface plasmon evanescent wave is produced. The value of the induced critical angle correlates with the permittivity of the media. The optical properties of the media can be obtained by calculating the amount of the angle shift. There are four optical configurations typically adopted for constructing the SPR, which are angular modulation, wavelength modulation, intensity modulation, and phase interrogation. In this study, angular modulation and phase information retrieval were chosen to pursue the further advancement of this technique. If the biomecular layers are taken as optical thin layers, the biomolecular interactions can be detected by SPR. Then the bio interaction curves were observed to analyze both the kinetic and static equilibrium reactions.
In a typical optical mechanism, design to arrive at precise incident angle control is most difficult since a tiny displacement associated with a large radius is needed. By extending the many years of optical system design experience within our research team and by introducing some innovative ideas, a design based on a paraboloidal lens-based surface plasmon resonance instrument has been created. In this new design, the angle control can be transformed to a stage-type movement control. The 0.001 degrees incident angle control can be easily obtained by today’s precision servomotor stage management. The SPR instrument was shown to be feasible. Furthermore, using such a system to measure the SPR critical angle shift during a bio-molecular reaction on a biochip; the bio-chemical reaction is analyzed by using the measurement results obtained. Not only the ELISA (Enzyme-Linked Immunosorbent Assays) interaction was done, but also BIA (Biomolecular Interaction Analysis) experiments with human IgG and anti-IgG were successfully measured. The SPR angle shift to the time plots were obtained. The reflection index variations were also achieved. The RIeq/C versus RIeq showed the kinetic affinity between human IgG and anti-IgG is 4.9380×10-5.
Since some of the kinetic affinity data measured is slightly different from the values obtained from previous studies, some ways to improve the measurement resolution were demonstrated. Besides, the design for phase interrogation SPR instrument developed by extending quadrature interferometer technique was also proposed and pursued. Suggestions on further improving its function were proposed as well.
Subjects
角度調制
拋物面鏡
分子親和力
表面電漿共振
相位調制
酵素連結免疫分析反應
Quadrature 干涉架構
生物分子交互作用分析
Angular Modulation
Bioaffinity
Surface Plasmon Resonance
Quadrature Inteferometer Technique
ELISA
Phase Interrogation
Paraboloidal Lens-based
BIA
Type
thesis
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