Conventional microbial cell cultivation techniques are typically labor intensive, low throughput, and poorlyparallelized, rendering them inefficient. The development of automated, modular microbial cell micro-cultivation systems, particularly those employing droplet microfluidics, have gained attention for their high-throughput, highly paralellized and efficient cultivation capabilities. Here, we report the development of a microbial microdroplet culture system (MMC), which is an integrated platform for automated, high-throughput cultivation and adaptive evolution of microorganisms. We demonstrated that the MMC yielded both accurate and reproducible results for the manipulation and detection of droplets. The superior performance of MMC for microbial cell cultivation was validated by comparing the growth curves of six microbial strains grown in MMC, conventional shake flasks or well plates. The highest incipient growth rate for all six microbial strains was achieved by using MMC. We also conducted an 18-day process of adaptive evolution of methanol-essential Escherichia coli strain in MMC and obtained two strains exhibiting higher growth rates compared with the parent strain. Our study demonstrates the power of MMC to provide an efficient and reliable approach for automated, high-throughput microbial cultivation and adaptive evolution.

中文翻译：

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微生物微滴培养系统 (MMC)：用于自动化、高通量微生物培养和适应性进化的集成平台。

传统的微生物细胞培养技术通常劳动密集型、产量低、并行性差，效率低下。自动化、模块化微生物细胞微培养系统的开发，特别是那些采用液滴微流体技术的系统，因其高通量、高度并行化和高效的培养能力而受到关注。在这里，我们报告了微生物微滴培养系统 (MMC) 的开发，该系统是用于微生物自动化、高通量培养和适应性进化的集成平台。我们证明了 MMC 为液滴的操纵和检测产生了准确和可重复的结果。通过比较在 MMC 中生长的六种微生物菌株的生长曲线，验证了 MMC 在微生物细胞培养方面的优越性能，常规摇瓶或孔板。所有六种微生物菌株的最高初始生长率都是通过使用 MMC 实现的。我们还在 MMC 中进行了为期 18 天的甲醇必需大肠杆菌菌株的适应性进化过程，并获得了与亲本菌株相比具有更高生长速率的两种菌株。我们的研究证明了 MMC 为自动化、高通量微生物培养和适应性进化提供有效和可靠方法的能力。