Microfluidic-chip Based Research On FISH Detection Technology

With the continuous development of society and industry,the incidence of a series of diseases(such as leukemia,also known as blood cancer)has continued to increase.Leukemia is divided into many types,and different types correspond to different treatment methods.Therefore,accurate determination and classification of the patient's disease type in prime time is particularly important for effective treatment of the condition.At present,the application of Fluorescence In Situ Hybridization(FISH)in the detection and typing of leukemia has achieved certain results and FISH is widely used in the fields of blood tumor diagnosis,solid tumor diagnosis,and prenatal diagnosis,etc.However,the application of domestic FISH detection technology faces some bottlenecks,including slow detection speed,high detection cost,and lack of automated equipment,which severely limits the promotion and application of FISH detection technology in the field of accurate detection.The microfluidic chip has the advantages of small size,high throughput,fast response,easy integration and automation.And the microfluidic chip as the core device to construct the FISH detection system is the ideal choice to solve the existing defects of the FISH detection technology(cumbersome processes,long detection time,excessive consumption of expensive reagents,insufficient detection flux,etc.).Therefore,designing a microfluidic chip that can be adapted to a fully automatic FISH staining system is a major subject of current application research.Based on FISH detection technology and detection process,this paper makes the following studies:(1)Understand the development status and trends of FISH detection technology,and analyze the problems of FISH detection technology in the application field,especially in the determination and typing of leukemia;(2)Complete the design and construction of the embedded slope etching device,including the design of the main control circuit,program design and writing,and the design of the etching fixture to realize the automation of the slope chip etching process;(3)Analyze the advantages and disadvantages of the existing equipment,combine the design requirements of the fully automatic FISH dyeing system,complete the design and optimization of the FISH chip based on the slope structure,and complete the design and construction of the pre-experiment platform;(4)Complete the design and research of the micro-dam array-based FISH chip based on the defects of the slope-based FISH chip,including the design and optimization of thechip structure,the formulation of the process route,the selection of manufacturing methods and materials,and the optimization of key process parameters,Determine the best specimen processing method,and complete the chip test;(5)Analyze the workflow design of the full-automatic FISH staining system and set up a semi-automatic FISH staining system,complete the chip FISH detection experiment verification,analyze and summarize the experimental results,and finally propose a summary and prospect.The chip designed in this paper satisfies the requirements of the full-automatic FISH staining system equipment while solving the main problems of the above-mentioned FISH detection technology in practical applications,and has the following innovations:(1)High detection efficiency,a single chip can detect four groups of samples at the same time,and the detection time is shortened to one sixth;(2)Closed trace reaction chamber;(3)The detection cost is low and the amount of reagents is small,especially the amount of expensive probes is reduced to one quarter;(4)Physical methods to achieve the separation and capture of target cells.