高通量自动化微生物微液滴进化培养与筛选技术及其装备化

Technology development and instrumentation of a high-throughput and automated microbial microdroplet culture system for microbial evolution and screening

摘要:

液滴微流控由于可以快速生成大量微液滴，并实现单个液滴独立的控制，每个液滴都可以作为独立的单元进行微生物培养，因此在微生物的高通量培养方面具有独特的应用优势。然而现有研究多停留在实验室搭建和使用阶段，存在操作要求高、影响因素多、缺乏自动化集成技术等关键问题，制约了液滴微流控技术在微生物研究中的应用。文中以解决液滴微流控技术用于微生物培养的装备化问题为目标，系统研究了微流控各单元模块的结构与功能，通过对液滴的发生、培养、检测、分割、融合、分选等多种操作的开发与集成，成功研制出了小型一体化、全自动高通量的微生物微液滴培养 (Microbial Microdroplet Culture system，MMC) 装备系统，可用于微生物的生长曲线测定、适应性进化、单因素多水平分析及代谢物检测等，为面向微生物菌种高效选育的进化培养和筛选提供了高通量仪器平台。

关键词:液滴微流控，微生物培养，高通量，微液滴，微生物进化，筛选

Abstract:

Since microdroplets are able to be generated rapidly in large amount and each droplet can be well controlled as an independent micro-cultivator, droplet microfluidic technology can be potentially used in the culture of microorganisms, and provide the microbial culture with high throughput manner. But its application mostly stays in the laboratory-level building and using for scientific research, and the wide use of droplet microfluidics in microbial technology has been limited by the key problems that the operation for microdroplets needs high technical requirements with wide affecting factors and the difficulties in integration of automatic microdroplet instrumentation. In this study, by realizing and integrating the complicated operations of droplet generation, cultivation, detection, splitting, fusion and sorting, we design a miniaturized, fully automated and high-throughput microbial microdroplet culture system (MMC). The MMC can be widely used in microbial growth curve test, laboratory adaptive evolution, single factor and multi-level analysis of microbial culture, metabolite detection and so on, and provide a powerful instrument platform for customized microbial evolution and screening aiming at efficient strain engineering.

Key words:droplet microfluidic, microbial culture, high-throughput, droplet, microbialevolution, screening