High-throughput tracking of single yeast cells in a microfluidic imaging matrix

Area of Science:

• Cellular dynamics and systems biology
• Microfluidics and live cell imaging
• Yeast genetics and signaling pathways

Background:

• Conventional live cell imaging lacks throughput and precise control for non-adherent cells.
• Studying yeast signaling requires robust cell tracking and large-scale analysis under perturbations.
• Existing methods hinder system-level investigations of cellular response dynamics.

Purpose of the Study:

• To present a high-throughput microfluidic imaging system for tracking single yeast cells.
• To enable programmable chemical control and multi-generational analysis.
• To facilitate system-level studies of signaling networks in non-adherent organisms.

Main Methods:

• Developed a microfluidic device with 128 independent imaging experiments.
• Immobilized yeast cells using mechanical clamping and agarose gel polymerization.
• Implemented programmable medium exchange via diffusion for precise chemical control.

Main Results:

• Achieved high-resolution imaging and robust single-cell tracking over multiple generations.
• Successfully analyzed yeast pheromone signaling across 8 genotypes and 16 conditions.
• Demonstrated lineage-dependent effects influencing cellular decision-making variability.

Conclusions:

• The microfluidic system enhances throughput and control for live cell imaging studies.
• Lineage history plays a significant role in cellular responses near decision thresholds.
• This technology is valuable for systems-level analysis of non-adherent cell populations.