Long-term single cell analysis of S. pombe on a microfluidic microchemostat array

Area of Science:

• Cell Biology
• Microbiology
• Biophysics

Background:

• Schyzosaccharomyces pombe is a key model organism for cell cycle research.
• Existing methods lack the capacity for high-throughput single-cell analysis of S. pombe growth and division.

Purpose of the Study:

• To develop a novel automated microfluidic platform for high-resolution, long-term single-cell analysis of S. pombe.
• To characterize cell cycle dynamics and size homeostasis in S. pombe under defined conditions.

Main Methods:

• Development of an automated microfluidic device for on-chip cultivation of S. pombe.
• Creation of an image processing pipeline to quantify cell length, division time, and elongation rate.
• Analysis of over 100,000 cell division events and reconstruction of multi-generational lineages.

Main Results:

• The platform enabled analysis of thousands of cells and tens of thousands of division events over 50 hours.
• Transient changes in cell length were observed during temperature shifts, depending on the cell cycle phase.
• Cell size homeostasis is regulated, with larger cells dividing more rapidly to normalize size in the next generation.

Conclusions:

• The microfluidic platform provides unprecedented detail for studying S. pombe cell cycle dynamics.
• This tool is valuable for investigating the molecular mechanisms of cell size control.
• The platform is broadly applicable to single-cell studies requiring large-scale, precise measurements and lineage tracking.