High-Throughput Single-Cell Cultivation on Microfluidic Streak Plates |
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| Authors: | Cheng-Ying Jiang Libing Dong Jian-Kang Zhao Xiaofang Hu Chaohua Shen Yuxin Qiao Xinyue Zhang Yapei Wang Rustem F Ismagilov Shuang-Jiang Liu Wenbin Du |
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| Institution: | University of California—Davis;aState Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China;bDepartment of Chemistry, Renmin University of China, Beijing, China;cDivision of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California, USA |
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| Abstract: | 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. |
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