Compartmentalized microchannel array for high-throughput analysis of single cell polarized growth and dynamics

Area of Science:

• Cell Biology
• Microfluidics
• Mycology

Background:

• Studying polarized growth is difficult due to cellular branching and uncontrolled environments in traditional assays.
• Advanced techniques are needed for precise control over temporal and spatial conditions during growth observations.

Purpose of the Study:

• To develop and demonstrate a high-performance microfluidic system for studying polarized growth.
• To enable detailed analysis of cellular heterogeneity, nuclear dynamics, and gene expression in filamentous fungi.

Main Methods:

• Development of a versatile microfluidic device with tunable environmental control.
• Utilizing the filamentous fungus Neurospora crassa as a model organism.
• High-resolution imaging and quantitative analysis of cellular processes at the single-hyphal compartment level.

Main Results:

• The microfluidic system provided enhanced temporal and spatial control for observing polarized growth.
• Direct visualization and analysis of cellular heterogeneity and nuclear dynamics were achieved.
• Quantitative gene expression dynamics were measured in response to nutrient changes with single-hyphal compartment resolution.

Conclusions:

• The developed microfluidic system offers a robust platform for studying polarized growth in filamentous fungi.
• The technology is adaptable for diverse biosystems exhibiting polarized growth, with potential applications in bioenergy and human health.
• This system significantly advances the ability to interrogate complex cellular behaviors under precisely controlled conditions.