Microfluidic chip-based single-cell liquid droplet high-throughput screening method

Abstract

The present invention discloses a microfluidic chip-based single-cell liquid droplet high-throughput screening method, which comprises: (1) generating a microbial single-cell liquid droplet on a microfluidic chip; (2) removing the unwrapped microbial liquid droplet; (3) fusing the cultured microbial single-cell liquid droplet and a small molecule compound biosensor on the microfluidic chip; and (4) making the biosensor respond to the metabolite in the fused liquid droplet, and carrying out fluorescence detection and sorting. According to the present invention, the single-cell liquid droplet culture is performed at the microfluidic chip, such that thousands of the micro-bioreactors are constructed on the chip so as to trace the growth characteristics of the microbial cells to be screened and screen; and through the liquid droplet fusion technology, the fluorescent labeling is performed on the microbial cell liquid droplet to be screened by the small molecule metabolite biosensor so as to screen the target extracellular product.

Description

technical field [0001] The invention relates to a cell screening method, in particular to a microfluidic chip-based single-cell droplet high-throughput screening method. Background technique [0002] In recent years, Microfluidic Chip (MC) technology has developed vigorously and gradually entered the era of industrialization. Microfluidic technology has the advantages of fast, high efficiency, and low consumption, and has broad prospects in many application fields such as biology, high-throughput drug screening, and disease diagnosis. Among them, Fluorescence-Activated Droplet Sorting (FADS) based on microfluidic technology has attracted great attention from researchers in the field of high-throughput screening. Its core is to wrap the sample to be tested and the fluorescent marker together in the droplet (water-in-oil), so as to realize the flow detection and sorting of the sample to be tested. In my country, researchers from the Tianjin Institute of Industrial Biotechnol...

AI Technical Summary

Problems solved by technology

However, these works still face the following technical defects in solving the high-throughput screening of industrial microorganisms: (1) Lack of single-cell culture technology, the culture process of microorganisms is performed in a deep-well plate system, and its throughput becomes the bottleneck of the overall screening efficiency (2) The fluorescent labeling system adopts the traditional double-enzyme coupling method, which has high cost, poor system compatibility, and limited application range

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