Research Progress in Microbial Single Cell Separation Methods

doi:10.13560/j.cnki.biotech.bull.1985.2022-1506

Abstract

Abstract:

Most microorganisms in nature are in an uncultured state，which are called “microbial dark matter”. With the development of technology innovations in microbial single-cell separation, significant progress has been made in applying new technologies and methods to the challenges of microbial pure-culture. These new separation and culture strategies will greatly promote the development of microbial resources research. Despite the increasing achievements related to metagenomics and genomics, the isolation and cultivation of single microbial cells are still essential to systematically study their ecological functions, genetic evolution, and so on. This paper mainly summarizes the principle and application of membrane diffusion culture, microfluidic sorting, fluorescence activated cell sorting, single cell Raman sorting, optical tweezers technology, micromanipulation technology and other single cell separation technologies currently used or under development, as well as their advantages and disadvantages in microbial single cell isolation and culture. At the same time, the paper also discusses development and application prospects of single-cell separation technologies in the future.

Key words: microbial dark matter, single cell separation, uncultured microbes, selective isolation of microorganisms

ZHANG Kun, YAN Chang, TIAN Xin-peng. Research Progress in Microbial Single Cell Separation Methods[J]. Biotechnology Bulletin, 2023, 39(9): 1-11.

Figures/Tables 4

References 71

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分离方法 Separation method		优点 Advantages	不足 Disadvantages
高通量分离方法	膜扩散培养法	原位培养微生物，增加可培养率	耗时久，工作量大
High throughput separation methods	微流控技术	高通量，结构简单，成本低	通用性及特异性低
	单细胞拉曼分选	分选效率高，无创，无标记	仪器要求较高，拉曼光谱比较弱
	荧光激活细胞分选	高通量，效率高，灵敏度高	成本高，仪器要求较高，损伤细胞
靶向分离方法	Live-FISH	靶向分离细胞效果较强	耗时久，细胞破坏性大，成活率低
Targeted separation methods	反向基因组学	靶向分离细胞，适用性广泛	需要依赖抗体标记和流式分选
其他方法	光镊技术	精度高、效率高	会损伤细胞，成本高
Other methods	显微操作技术	适用范围广，可分离选定细胞	效率低，容易损伤细胞

分离方法 Separation method		优点 Advantages	不足 Disadvantages
高通量分离方法	膜扩散培养法	原位培养微生物，增加可培养率	耗时久，工作量大
High throughput separation methods	微流控技术	高通量，结构简单，成本低	通用性及特异性低
	单细胞拉曼分选	分选效率高，无创，无标记	仪器要求较高，拉曼光谱比较弱
	荧光激活细胞分选	高通量，效率高，灵敏度高	成本高，仪器要求较高，损伤细胞
靶向分离方法	Live-FISH	靶向分离细胞效果较强	耗时久，细胞破坏性大，成活率低
Targeted separation methods	反向基因组学	靶向分离细胞，适用性广泛	需要依赖抗体标记和流式分选
其他方法	光镊技术	精度高、效率高	会损伤细胞，成本高
Other methods	显微操作技术	适用范围广，可分离选定细胞	效率低，容易损伤细胞