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Microdroplet-based system for culturing of environmental microorganisms using FNAP-sort
Kanako Saito1,2, Yuri Ota1,2, Dieter M Tourlousse2
1Department of Life Science and Medical Bioscience, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo, 162-0056, Japan.
This study introduces an integrated microfluidic system for efficiently isolating and culturing rare environmental microbes. The technology combines droplet microfluidics with fluorescent nucleic acid probe sorting (FNAP-sort) to recover growth-positive droplets, enabling high-purity microbial cultures.
Area of Science:
- Microbiology
- Biotechnology
- Environmental Science
Background:
- Droplet microfluidics enhances microbial culturing efficiency.
- Identifying and manipulating individual growth-positive droplets remains a challenge.
- A significant proportion of environmental microorganisms are currently uncultured.
Purpose of the Study:
- To develop an end-to-end workflow for isolating and culturing environmental microbes using droplet microfluidics.
- To overcome technical challenges in identifying and manipulating growth-positive droplets.
- To enable the culturing of rare and uncultured microorganisms.
Main Methods:
- Integration of microfluidic droplet technology with on-chip "fluorescent nucleic acid probe in droplets for bacterial sorting" (FNAP-sort).
- Utilizing FNAP-sort for recovery of growth-positive droplets.
- Employing droplet microdispensing for microbial isolation and culturing.
Main Results:
- Demonstrated an end-to-end workflow for microbial isolation and culturing.
- Successfully yielded high-purity cultures of rare microorganisms from a complex environmental microbiome.
- The system utilizes off-the-shelf, commercially available equipment.
Conclusions:
- The developed technique provides a robust method for culturing environmental microbes.
- The system's reliance on accessible equipment facilitates widespread adoption.
- This workflow can significantly contribute to culturing uncultured microorganisms across diverse biomes.
