Research And Application Of Microfluidic Chip And Automated Instrumentation Based On Immunofluorescence Detection

Various infectious diseases,such as historical smallpox to SARS,and the new crown pneumonia(COVID-19),which began spreading in 2019,pose a serious threat to global public health security,causing enormous losses in all aspects of human life and health,as well as global economy and society.For infectious disease detection,polymerase chain reaction(PCR),based on the principle of nucleic acid amplification,is one of the accepted standard method.Specific immune reactions based on antigen-antibodies can also enable the rapid detection of infectious diseases.The research in this paper focuses on two aspects.On the one hand,combining the immunoassay method with a microfluidic chip,with the flexible fluidic drive of the microfluidic chip,helps to simplify the sample pre-treatment steps in the assay process.A microfluidic chip based on an elastic reaction chamber was constructed for the joint detection of multiple indicators.The elastic reaction chamber is a storage and incubation chamber for reaction samples,and at the same time,it can realize simplified fluidic driving by its elastic deformation.In particular,the realization of parallel fluid driving for multi-target detection significantly reduces the complexity of the chip and the device.Ultimately,several steps such as fluid quantification drive,sample mixing and dilution,and paper strip detection were integrated with each other,thus realizing the combined detection of multiple indicators.Another type is a multi-channel chip fully integrated immunoassay microfluidic chip.Another type of microfluidic chip is the multi-channel fully integrated immunoassay microfluidic chip.Based on the design idea of integrating multiple functional modules and a multi-functional reaction chamber,the POCT microfluidic chip was constructed.The microfluidic chip can automatically and continuously complete multiple steps required for nodule detection: long time sample incubation,quantitative fluid drive,red blood cell filtration,strip chromatography,and strip detection,realizing the "sample in-result out" detection process.Compared with the traditional method,it requires less testing equipment and simple manual steps,which is conducive to simple,efficient and convenient nodule detection.On the other hand,researching innovative methods and instruments of immunofluorescence detection based on Opto-mechatronics,especially highthroughput and automated immunofluorescence detection technology and instruments,can achieve efficient and reliable immunofluorescence detection.The study optimized the scheduling strategy of immunofluorescence highthroughput detection based on the turntable method and obtained the ideal system detection efficiency.The study optimized the fluorescence signal filtering algorithm and combined with the frequency domain filtering method to improve the signal-to-noise ratio of fluorescence signal and enhance the key detection performance.In addition,the search and identification algorithm was optimized for the test and control lines of immunochromatographic paper strips,which significantly improves the accuracy of the T-line and C-line recognition.In particular,the accuracy of T-line identification of weakly positive samples has been improved,and the sensitivity and reproducibility of assay have been improved.Finally high-performance,high-throughput immunofluorescence detection was achieved.