Protocol to perform dynamic microfluidic single-cell cultivation of C. glutamicum

Area of Science:

• Microbiology
• Biotechnology
• Biochemical Engineering

Background:

• Dynamic single-cell cultivation is crucial for understanding microbial physiology.
• Existing methods often lack the resolution to observe rapid cellular responses.
• Corynebacterium glutamicum is an important industrial microorganism.

Purpose of the Study:

• To present a protocol for a novel microfluidic chip for dynamic single-cell cultivation.
• To enable real-time observation of Corynebacterium glutamicum responses to environmental stimuli.
• To provide a versatile system adaptable to other cell-wall containing organisms.

Main Methods:

• Design and fabrication of a polydimethylsiloxane (PDMS)-based microfluidic chip.
• Establishment of flow profiles and biological preparation for cultivation.
• Time-lapse imaging to monitor cellular behavior under dynamic conditions.

Main Results:

• Successful dynamic microfluidic single-cell cultivation of Corynebacterium glutamicum.
• Observation of rapid cellular responses to environmental changes within seconds.
• Demonstration of the system's adaptability for other microorganisms and soluble factors.

Conclusions:

• The developed PDMS microfluidic chip offers a powerful tool for dynamic single-cell studies.
• This protocol facilitates high-resolution analysis of microbial responses to fluctuating environments.
• The system has broad applicability in microbial research and biotechnology.