Development of a facile droplet-based single-cell isolation platform for cultivation and genomic analysis in microorganisms
文献类型： 外文期刊
作者： Zhang, Qiang 1 ; Wang, Tingting 1 ; Zhou, Qian 1 ; Zhang, Peng 1 ; Gong, Yanhai 1 ; Gou, Honglei 1 ; Xu, Jian 1 ; Ma, Bo 1 ;
作者机构： 1.Chinese Acad Sci, CAS Key Lab Biofuels, Single Cell Ctr, Qingdao 266101, Peoples R China
2.Chinese Acad Sci, Shandong Key Lab Energy Genet, Qingdao Inst Bioenergy & Bioproc Technol, Qingdao 266101, Peoples R China
3.Chinese Acad Fishery Sci, Key Lab Sustainable Dev Marine Fisheries, Minist Agr, Yellow Sea Fisheries Res Inst, Qingdao 266071, Peoples R China
期刊名称：SCIENTIFIC REPORTS （ 影响因子：4.379； 五年影响因子：5.133 ）
ISSN： 2045-2322
年卷期： 2017 年 7 卷
页码：
收录情况： SCI
摘要： Wider application of single-cell analysis has been limited by the lack of an easy-to-use and low-cost strategy for single-cell isolation that can be directly coupled to single-cell sequencing and single-cell cultivation, especially for small-size microbes. Herein, a facile droplet microfluidic platform was developed to dispense individual microbial cells into conventional standard containers for downstream analysis. Functional parts for cell encapsulation, droplet inspection and sorting, as well as a chip-totube capillary interface were integrated on one single chip with simple architecture, and control of the droplet sorting was achieved by a low-cost solenoid microvalve. Using microalgal and yeast cells as models, single-cell isolation success rate of over 90% and single-cell cultivation success rate of 80% were demonstrated. We further showed that the individual cells isolated can be used in high-quality DNA and RNA analyses at both gene-specific and whole-genome levels (i.e. real-time quantitative PCR and genome sequencing). The simplicity and reliability of the method should improve accessibility of single-cell analysis and facilitate its wider application in microbiology researches.
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