Recently, the team led by Zhu Daochen from Jiangsu University Student Material Energy Research Institute published a research paper titled "Single cell mica droplet screening microfluidic system enables high-throughput isolation and * * * of lignin grading bacteria from the terminal gut" in Insect Science《Microfluidic system based on single-cell microliter droplets to assist in high-throughput separation and cultivation of termite intestinal lignin degrading bacteria》. The study willHigh throughput micro upgraded droplet culture omics system (MISS cell)Applied to the study of termite gut microbiota, it provides a powerful methodological tool for exploring difficult to cultivate microbial functional resources.

1、 Research background:

Lignin is one of the key components of plant biomass, and its efficient degradation and conversion are the difficulties in biomass resource utilization. As a natural lignocellulose degradation system, termite gut contains a large amount of uncultured microbial resources, especially in lignin degradation, which has great potential. However, the traditional agar plate method is difficult to comprehensively obtain the microbial diversity, which seriously restricts the development and utilization of related microbial resources.

2、 Research methods:

This study focuses on higher termites（Nasutiterms tiantongensis）For the sample, the intestinal microbiota suspension was cultured using the following two methods:

Traditional agar plate method:
Using lignin assingleSolid culture medium for carbon source separation by streaking.

● MISS cell method:
Based on droplet microfluidic technology, the gut microbiota is monodisperse into independent micro upgraded droplets.Each droplet is a closed and independent liquid confined space, effectively avoiding competition between colonies and providing an ideal environment for the growth of weak and difficult to cultivate strains.

The study used 16S rRNA gene amplicon sequencing technology to systematically compare and analyze the diversity of strains obtained by two methods.

3、 Research results:

1. Significant improvement in separation efficiency：

MISS Cell System Separated Together477 piecesLignin degrading bacterial colonies, while traditional methods only obtain73 of themThe number of bacterial colonies increased by about 6.5 times, demonstrating its significant advantage in microbial isolation efficiency.

2. Enhanced ability to capture species diversity：

① Two methods obtained a total of 97 operational taxonomic units (OTUs). Among them, there are 31 MISS cell methods (31.96%) and 20 traditional methods (20.62%).

② The total number of OTUs obtained by the MISS cell system is 16.7% higher than traditional methods.

③ Compared with the traditional agar plate method, the microbial community obtained by MISS cell culture has higher diversity, indicating that this system is an effective tool for obtaining uncultured microbial resources.

3. Differences in community structure:
Both methods successfully captured major bacterial groups such as Proteobacteria and Firmicutes, but at the genus level, MISS cells exhibited a richer species composition, successfully obtaining multiple known functional bacterial genera including Streptomyces and Pseudomonas, as well as a large number of unclassified microbial groups.

Research conclusions:
This study indicates that,MISS Cell SystemThrough its unique micro droplet culture system, lignin degrading bacteria in termite intestines can be more efficiently and comprehensively isolated, effectively breaking through the technical limitations of traditional culture methods. This technology provides reliable technical support for resource mining and functional development of difficult to cultivate microorganisms in complex environmental samples.

About MISS Cell

The Single cell Microlite droplet Culture Omics System (MISS cell culture omics) is a miniaturized high-throughput single-cell culture and sorting equipment developed based on droplet microfluidics technology. It can process about 5000-20000 droplets (500-4000 monoclonal antibodies) in a single run experiment. After droplet generation, it is stored in a highly permeable pipeline for incubation (0-60 days), and finally detected and sorted by optical signals (OD, fluorescence, chemiluminescence, etc.) to achieve separation and cultivation of environmental bacterial communities at the single-cell level. The target droplets are sorted and stored in a porous plate.

Four Major Advantages

1Droplet system

The core advantage of MISS cell lies in its unique micro upgrade droplet system design, where micro droplets are independent reactors that achieve higher species richness compared to traditional agar plate separation and cultivation methods

2Efficient Separation and Cultivation

MISS cells create independent and uniform droplet culture environments for each microbial cell, which can effectively avoid cross contamination and competition inhibition between colonies, providing optimal growth conditions for low abundance and difficult to cultivate strains

3High degree of automation

After the MISS cell is loaded onto the sample machine, the entire process of preparation, incubation, detection, sorting, and target droplet collection is automated

4Rich application scenarios

MISS cells are used for high-throughput screening of bacteria, yeast, mold, actinomycetes, algae, and other microorganisms; Isolation and screening of environmental microorganisms; Cultivation and screening of microorganisms in special gas environments such as anaerobic bacteria and nitrogen fixing bacteria; Enzyme evolution, antibody screening, drug screening, synthetic biology and other scenarios can be efficiently applied