title: High-Throughput Single-Cell Cultivation on Microfluidic Streak Plates.
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.
authors: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
authors: Cheng-Ying
authors: CY
authors: Jiang
authors: Department of Chemistry, Renmin University of China, Beijing, China.
authors: Libing
authors: L
authors: Dong
authors: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
authors: Jian-Kang
authors: JK
authors: Zhao
authors: Department of Chemistry, Renmin University of China, Beijing, China.
authors: Xiaofang
authors: X
authors: Hu
authors: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
authors: Chaohua
authors: C
authors: Shen
authors: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
authors: Yuxin
authors: Y
authors: Qiao
authors: Department of Chemistry, Renmin University of China, Beijing, China.
authors: Xinyue
authors: X
authors: Zhang
authors: Department of Chemistry, Renmin University of China, Beijing, China.
authors: Yapei
authors: Y
authors: Wang
authors: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California, USA.
authors: Rustem F
authors: RF
authors: Ismagilov
authors: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China wenbin@im.ac.cn liusj@im.ac.cn.
authors: Shuang-Jiang
authors: SJ
authors: Liu
authors: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China wenbin@im.ac.cn liusj@im.ac.cn.
authors: Wenbin
authors: W
authors: Du
mesh_terms: D004926:Escherichia coli / Q000254:growth & development* / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D011084:Polycyclic Aromatic Hydrocarbons / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000254:growth & development* / Q000378:metabolism
chemical_list: D011084:Polycyclic Aromatic Hydrocarbons