Abstract:Most microorganisms are still unculturable in vitro, therefore studies of the “microbial dark matter” have great significance for investigating the evolution of microorganisms and making the most of microbial resources. Research on the isolation and cultivation of uncultivated microorganisms has become a hot research topic in the world. In this paper, we introduce the principle and development of different single cell sorting techniques, such as micromanipulation, fluorescence activated cell sorting, microfluidic cell sorting, optical tweezer, laser-induced forward transfer, to isolate and culture microorganisms. According to the advantages and disadvantages of each sorting technology, we mainly describe the advantages of laser-induced forward transfer and prospect the application of this technology to the research of uncultivated microorganisms.