Study On Peptide Nucleic Acid GeneChip Detection System Based On Surface Plasmon Resonance

Most of the biomolecular recognition process operate at interfaces. SPR(Surface Plasmon Resonance)technology is used to measure biomolecular interactions in real time with high sensitivity and without labels. SPR imaging provides great convenience of monitoring adsorption across a large area of sensor surface with high resolution.Peptide nucleic acid (PNA) is a nucleic acid mimics design by CAD, in which the sugar phosphate backbone of natural nucleic acid is replaced by a synthetic peptide backbone formed by N-(2-aminoethyl)-glycine units. PNA can recognize DNA and RNA specifically and PNA-DNA, PNA-RNA complexes are more stable than corresponding DNA-DNA, DNA-RNA. Also, PNA is superior to DNA and RNA in recognizing single nucleotide polymorphism. These characteristics are very suiltable for clinical requirements, has the possibility of main hybridization mode in the fulture.Gold nanoparticle has been applied in the trace technique of life science for a long time. Due to its unique physical, chemical quality and biocompatibility, gold nanoparticle has also been widely used in GeneChip detection technology recently.In particular, as the discrete nanogold has no quality of adsorption, which used as signal report material is regardesd as a significant development in genechip detection technology.This work is supported by the NSFC (National Natrual Science Funding of China).The purpose is to design and manufacture a high-throughput SPR system. Here we give the theoretical analysis of surface plasmon wave, dipict the PNA and nanogold principle and applications.This dissertation describes the development of high-throughput techniques based on SPR imaging to study hybridizaion between PNA and DNA .In this experiment, PNA was immobilized on nanoglod surface. It is of great practical significance to develop a home-made SPR detection system based on PNA probe and nanogold particles. We give a research of SPR principle on film optics from the point of view of external field and internal field, present the resonance model based on SPW and ATR wave. The theoretical foundation is basis of design and fabrication of SPR detection system and SPR biosensor.The home-made high-throughput SPR detection system, which includes optic unit, biosense unit, flow unit, image acquisition-processing unit. It is fabricated on angular interrogation, which has high angular flexible and fulifill wide range measurement. Realized by laser beam expand methods.We also show the method of FIA (Flow Injection Analysis) and realized through peristaltic pump, six-way valve and flow cell.CCD array detector, frame-grabber card, image acquisition software constitute the image acquisition unit. The image processing software can store, display, process GeneChip image. Image preprocessing, half-automatic gridding or automatic gridding (image segmentation) are important tasks before data analysis. Orientation information measurement of image and wavelet theoretics algorithms are employed in GeneChip...