The Integration Of A Micro-zone Reflectometric Interference Spectroscopy Detection System And Its Application In The Detection Of Molecular Interactions

After years of development,Reflectometric Interference Spectroscopy(RIfS)has become a mature detection technology,which plays an increasingly important role in the fields of physics,chemistry,biology,medicine,public safety and environmental detection.With the development of thin film technology,a variety of nanostructured thin films with good optical properties have become a research hotspot.Many biomolecular interactions can be completed in nanostructures of thin films and these reactions can be detected in real time by optical means.The combination of RIfS and mature thin film technology has a good application prospect and would play a key role in the biomedical field.Based on the original RIfS system,a micro-zone reflectometric interference spectrum detection system is constructed in this paper to realize the real-time detection of the reflectometric interference spectrum in the micro scale.The signal intensity,signal-to-noise ratio and stability are optimized.The response of nanostructured films under different dielectric conditions is explored and the real-time monitoring of the interaction process of different molecules in nanopores is realized.The above works broaden the idea for the improvement of RIfS technology and the realization of real-time detection of molecular interaction.The specific work contents are as follows:1.The inverted microscope was used to construct the micro-zone reflectometric interference spectroscopy system.The system was optimized after the improvement and adjustment of field of view aperture,optical fiber collimator,light source,etc.The stability,sensitivity and signal-to-noise ratio of the system were improved.Aluminum alloy sample tank and acrylic flow cell were designed and manufactured to meet the experimental requirements.In situ real-time analysis of micro-zone reflection interference spectrum was realized.2.The ordered porous polystyrene film was combined with the micro-zone reflectometric interference spectroscopy system.The response of ordered porous polystyrene film to the refractive index of different media was studied.The advantage of micro-zone method in real-time detection with high signal-to-noise ratio was proved through concentration gradient experiment.By monitoring the optical thickness of ordered porous polystyrene film in different solvents,the slight swelling effect of ordered porous polystyrene film in lower alcohols was confirmed3.The binding process of staphylococcal protein A(SPA)and immunoglobulin G(IgG)in nanopores was investigated by using ordered porous polystyrene film as sensor.The change of IgG activity after high temperature treatment was analyzed with silica colloidal crystal film as substrate.In addition,the difference between tracing diffraction peak and tracing interference peak in detecting protein adsorption behavior was studied and analyzed,which proved that tracing interference peak is a more sensitive method.The real-time detection and analysis of the molecular interaction process in the porous film were realized by using the micro-zone reflectometric interference spectroscopy detection system.