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High throughput Single-cell Cultivation on Microfluidic Streak Plates

Jiang, Cheng-Ying and Dong, Libing and Zhao, Jian-Kang and Hu, Xiaofang and Shen, Chaohua and Qiao, Yuxin and Zhang, Xinyue and Wang, Yapei and Ismagilov, Rustem F. and Liu, Shuang-Jiang and Du, Wenbin (2016) High throughput Single-cell Cultivation on Microfluidic Streak Plates. Applied and Environmental Microbiology, 82 (7). pp. 2210-2218. ISSN 0099-2240. PMCID PMC4807504. https://resolver.caltech.edu/CaltechAUTHORS:20160216-130203405

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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.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1128/AEM.03588-15DOIArticle
http://aem.asm.org/content/82/7/2210PublisherArticle
http://aem.asm.org/content/82/7/2210/suppl/DCSupplementalPublisherSupplemental Material
ORCID:
AuthorORCID
Ismagilov, Rustem F.0000-0002-3680-4399
Du, Wenbin0000-0002-7401-1410
Additional Information:© 2016 American Society for Microbiology. Received 4 November 2015; Accepted 19 January 2016; Accepted manuscript posted online 5 February 2016. We thank Natasha Shelby and Liang Ma for help in editing the manuscript. We declare no conflict of interest. This work is supported by the National Natural Science Foundation of China (31230003, 21205134, 21377161, and 31470221), the program of China Ocean Mineral Resources R&D Association (DY125-15-R-02), the Strategic Priority Research Program of Chinese Academy of Sciences (XDB15040102), and the National High Technology Research and Development Program of China (2012AA092103).
Funders:
Funding AgencyGrant Number
National Natural Science Foundation of China31230003
National Natural Science Foundation of China21205134
National Natural Science Foundation of China21377161
National Natural Science Foundation of China31470221
Program of China Ocean Mineral Resources R&D AssociationDY125-15-R-02
Chinese Academy of SciencesXDB15040102
National High Technology Research and Development Program of China2012AA092103
Issue or Number:7
PubMed Central ID:PMC4807504
Record Number:CaltechAUTHORS:20160216-130203405
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20160216-130203405
Official Citation:High-Throughput Single-Cell Cultivation on Microfluidic Streak Plates Cheng-Ying Jiang, Libing Dong, Jian-Kang Zhao, Xiaofang Hu, Chaohua Shen, Yuxin Qiao, Xinyue Zhang, Yapei Wang, Rustem F. Ismagilov, Shuang-Jiang Liu, and Wenbin Du Appl. Environ. Microbiol. April 2016 82:7 2210-2218; Accepted manuscript posted online 5 February 2016, doi:10.1128/AEM.03588-15
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:64512
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:17 Feb 2016 22:28
Last Modified:09 Mar 2020 13:18

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