Fabrication Of 2D Micro/nanobowl Arrays And Their Biochip Application

Biochip technology has broad application prospects in biomedical engineering such as cell screening,protein expression,gene sequencing,and nucleic acid detection,and has become a research hotspot in related fields at home and abroad.Cell array is an important platform for high-throughput detection and screening of biochips.It is widely used in high-throughput screening and single-cell sequencing research in biopharmaceutical and clinical fields,and has gradually become a hot spot in the field of biochips.Among them,the micro/nanobowl array has good application value in the cell array chip due to its unique physical and chemical properties and structural characteristics.In this paper,we explore the micro/nanosphere array structure design,preparation and modification methods,and controls the micro-structure by deposition and etching parameters,in order to achieve microbial single cell array screening and precise positioning.The goal is to achieve high-throughput detection and screening of single cells.In this paper,we have studied a large-area two-dimensional micro/nanosphere array as a platform for microarray chips,using microbial cells as research objects to realize cell detection and screening techniques.In order to verify the application value of the micro/nanobowl array in the cell array chip,the fluorescent microspheres were firstly used for pre-experimental placement in the micro/nanobowl.The laser confocal microscope and electron microscope were used to characterize and found that the regularly ordered fluorescent microspheres array were formed on the micro/nanobowl array.Then,cocci(S.aureus,S.suis)bacilli(E.coli,S.flexneri)and spores(B.anthracis spores)were used as the representative,and the intravital implantation experiment was carried out to study the influence of bacteria status,bacteria shape and bacteria size.When a suspension of droplets containing monodisperse bacteria is applied to the array of micro/nanobowl,if the size of the bacteria is smaller than the size of the bowl,the bacteria enter the bowl and form a regular array of single cells.If the size of the bacteria is larger than the size of the micro/nanobowl,such as chain S.suis,long bacillus,bacteria are difficult to enter the micro/nanobowl array to form an array,and the bacteria are placed horizontally on the wall of the micro/nanobowl array.Some bacilli can be vertically inserted into the micro/nanobowl at different angles to form a bacterial array.With fluorescent labeling and confocal microscopy,it is possible to efficiently locate and screen individual bacteria on large-area substrates loaded with millions of bacteria.Combined with micromanipulation techniques,efficient bacterial selection can be achieved.This study demonstrates a simple,accurate,and cost-effective microbial high-throughput detection and screening technique based on micro/nanosphere arrays.