Single-cell microliter-droplet screening system (MISS Cell): An integrated platform for automated high-throughput microbial monoclonal cultivation and picking

被引:9
|
作者
Jian, Xingjin [1 ,2 ]
Guo, Xiaojie [1 ,2 ]
Cai, Zhengshuo [1 ,2 ]
Wei, Longfeng [3 ]
Wang, Liyan [4 ]
Xing, Xin-hui [1 ,2 ,5 ]
Zhang, Chong [1 ,2 ,5 ]
机构
[1] Tsinghua Univ, Inst Biochem Engn, Dept Chem Engn, Beijing 100084, Peoples R China
[2] Tsinghua Univ, Key Lab Ind Biocatalysis, Minist Educ, Beijing, Peoples R China
[3] Guizhou Univ, Inst Agrobioengn, Coll Life Sci, Guiyang, Peoples R China
[4] Luoyang TMAXTREE Biotechnol Co Ltd, Luoyang, Peoples R China
[5] Tsinghua Univ, Ctr Synthet & Syst Biol, Beijing, Peoples R China
基金
中国国家自然科学基金;
关键词
automated operations; colony picking; high-throughput; microbial cultivation; MISS Cell; single-cell microliter-droplet screening system; TEMPERATURE PLASMA ARTP; BACTERIAL-GROWTH; PLATE; CULTURE; MICROFLUIDICS; MEDIA;
D O I
10.1002/bit.28300
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Solid plates have been used for microbial monoclonal isolation, cultivation, and colony picking since 1881. However, the process is labor- and resource-intensive for high-throughput requirements. Currently, several instruments have been integrated for automated and high-throughput picking, but complicated and expensive. To address these issues, we report a novel integrated platform, the single-cell microliter-droplet screening system (MISS Cell), for automated, high-throughput microbial monoclonal colony cultivation and picking. We verified the monoclonality of droplet cultures in the MISS Cell and characterized culture performance. Compared with solid plates, the MISS Cell generated a larger number of monoclonal colonies with higher initial growth rates using fewer resources. Finally, we established a workflow for automated high-throughput screening of Corynebacterium glutamicum using the MISS Cell and identified high glutamate-producing strains. The MISS Cell can serve as a universal platform to efficiently produce monoclonal colonies in high-throughput applications, overcoming the limitations of solid plates to promote rapid development in biotechnology.
引用
收藏
页码:778 / 792
页数:15
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