Jiang Cheng-Ying, Dong Libing, Zhao Jian-Kang, Hu Xiaofang, Shen Chaohua, Qiao Yuxin, Zhang Xinyue, Wang Yapei, Ismagilov Rustem F, Liu Shuang-Jiang, Du Wenbin
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Department of Chemistry, Renmin University of China, Beijing, China.
Appl Environ Microbiol. 2016 Feb 5;82(7):2210-8. doi: 10.1128/AEM.03588-15.
This paper describes the microfluidic streak plate (MSP), a facile method for high-throughput microbial cell separation and cultivation in nanoliter sessile droplets. The MSP method builds upon the conventional streak plate technique by using microfluidic devices to generate nanoliter droplets that can be streaked manually or robotically onto petri dishes prefilled with carrier oil for cultivation of single cells. In addition, chemical gradients could be encoded in the droplet array for comprehensive dose-response analysis. The MSP method was validated by using single-cell isolation of Escherichia coli and antimicrobial susceptibility testing of Pseudomonas aeruginosa PAO1. The robustness of the MSP work flow was demonstrated by cultivating a soil community that degrades polycyclic aromatic hydrocarbons. Cultivation in droplets enabled detection of the richest species diversity with better coverage of rare species. Moreover, isolation and cultivation of bacterial strains by MSP led to the discovery of several species with high degradation efficiency, including four Mycobacterium isolates and a previously unknown fluoranthene-degrading Blastococcus species.
本文介绍了微流控划线平板（MSP），这是一种在纳升静态液滴中进行高通量微生物细胞分离和培养的简便方法。MSP方法基于传统的划线平板技术，通过使用微流控装置生成纳升液滴，这些液滴可以手动或通过机器人划线到预先填充有载油的培养皿上以培养单细胞。此外，可以在液滴阵列中编码化学梯度以进行全面的剂量反应分析。通过对大肠杆菌进行单细胞分离和对铜绿假单胞菌PAO1进行抗菌药敏试验，验证了MSP方法。通过培养一个降解多环芳烃的土壤群落，证明了MSP工作流程的稳健性。在液滴中培养能够检测到最丰富的物种多样性，并更好地覆盖稀有物种。此外，通过MSP分离和培养细菌菌株导致发现了几种具有高降解效率的物种，包括四种分枝杆菌分离株和一种以前未知的降解荧蒽的芽球菌属物种。
