| 论文题目: | High throughput Single-cell Cultivation on Microfluidic Streak Plates |
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| 作者: | Jiang Cheng-Ying, Dong Libing, Zhao Jian-Kang, Hu Xiaofang, Shen Chaohua, Qiao Yuxin, Zhang Xinyue, Wang Yapei, Ismagilov Rustem F, Liu Shuang-Jiang*, and Du Wenbin*. |
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| 刊物名称: | Appl Environ Microbiol |
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| 年份: | 2016 |
| 影响因子: | 4.359 |
| 论文下载: | http://aem.asm.org/content/early/2016/02/01/AEM.03588-15 |
| 摘要: | 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 conventional streak plate technique 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 gradient could be encoded in the droplet array for comprehensive dose-response analysis. The MSP method was validated using single-cell isolation of Escherichia coli and antimicrobial susceptibility testing of Pseudomonas aeruginosa PAO1. The robustness of MSP workflow was demonstrated by cultivating a soil community that degrades polycyclic aromatic hydrocarbons (PAH). Cultivation in droplets enabled detection of the richest species diversity with better coverage of the rare species. Moreover, isolation and cultivation of bacterial strains with 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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