Developing a low-cost milliliter-scale chemostat array for precise control of cellular growth

Area of Science:

• Microbiology
• Biotechnology
• Synthetic Biology

Background:

• Multiplexed milliliter-scale chemostats are valuable tools for cell physiology studies and evolution experiments.
• Accurate characterization of flow rate parameters (accuracy, precision, operational range) is lacking in existing chemostat devices.
• Chemostats enable controlled nutrient limitation by pumping fresh medium and removing effluent at equal rates.

Purpose of the Study:

• To develop and characterize a multiplexed milliliter-scale chemostat system for precise control of cellular growth.
• To assess the accuracy, precision, and operational range of flow rates in the developed chemostat array.
• To demonstrate the utility of the chemostat system for accurate and precise quantification of population phenotypes.

Main Methods:

• Repurposing a published multiplexed culturing device to create a milliliter-scale chemostat array.
• Independently controlling flow rates across eight chambers for a wide range of population doubling times (3-13 hours).
• Evaluating flow rate accuracy and precision without expensive feedback systems.

Main Results:

• Flow rates were highly precise, with a maximal coefficient of variation below 3% across chambers.
• Average flow rates were accurate, with deficits of 3%-6% for 13-h and 0.6%-1.0% for 3-h doubling times, primarily due to evaporation.
• The device successfully enabled accurate and precise quantification of population phenotypes.

Conclusions:

• Precise control of cellular growth was achieved using a low-cost, milliliter-scale chemostat array.
• The high precision of the chemostat system reduces measurement errors in biological processes.
• This technology offers a cost-effective solution for microbial research and evolution experiments.