Long-Term and Programmable Bacterial Subculture in Completely Automated Microchemostats

Area of Science:

• Microbiology
• Biotechnology
• Bioengineering

Background:

• Conventional macroscale chemostats have limitations in controlling microbial growth and population dynamics.
• Simultaneous continuous growth and active population control are challenging due to microbial metabolic communication.

Purpose of the Study:

• To develop a novel microchemostat for improved study of microorganism growth behavior.
• To enable simultaneous continuous growth, active population control, and dynamic physicochemical stimulation.
• To achieve long-term bacterial subculture with precise population manipulation and ultralow reagent consumption.

Main Methods:

• Development of a microfluidic device enabling reversible bacterial isolation and continuous chemical refreshment.
• Implementation of automated and programmed control of bacterial growth and subculture conditions.
• Integration of in situ measurement and feedback control for bacterial growth and population dynamics.

Main Results:

• Demonstrated manipulation of bacterial populations from single cells to ultrahigh densities.
• Achieved long-term subculture (720 hours) with ultralow reagent consumption, the longest reported microchemostat culture.
• Successfully performed various subculture programming modes sequentially within a single microchemostat.

Conclusions:

• The novel microchemostat offers a powerful platform for studying microbial growth and population control.
• It enables precise, automated, and long-term microbial cultivation with unprecedented control.
• The system is broadly applicable to diverse microbial studies including nutrient optimization, genetic induction, and evolutionary adaptation.