A microfluidic system for studying ageing and dynamic single-cell responses in budding yeast

• Published in: PloS one Vol. 9; no. 6; p. e100042
• Main Authors: Crane, Matthew M, Clark, Ivan B N, Bakker, Elco, Smith, Stewart, Swain, Peter S
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 20.06.2014; Public Library of Science (PLoS)
• Subjects: Aging; Baking yeast; Biology; Biology and Life Sciences; Cell cycle; Cell Division; Computational fluid dynamics; Daughters; Decision making; Design; Engineering and Technology; Fluid flow; Gene expression; Injections; Kinases; Localization; Measuring instruments; Microfluidic Analytical Techniques - methods; Microfluidics; Physical Sciences; Proteins; Research and Analysis Methods; Saccharomyces cerevisiae; Saccharomycetales - cytology; Single-Cell Analysis - methods; Stochasticity; Stress response; Time Factors; Transcription factors; Yeast
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0100042

Recognition of the importance of cell-to-cell variability in cellular decision-making and a growing interest in stochastic modeling of cellular processes has led to an increased demand for high density, reproducible, single-cell measurements in time-varying surroundings. We present ALCATRAS (A Long-term Culturing And TRApping System), a microfluidic device that can quantitatively monitor up to 1000 cells of budding yeast in a well-defined and controlled environment. Daughter cells are removed by fluid flow to avoid crowding allowing experiments to run for over 60 hours, and the extracellular media may be changed repeatedly and in seconds. We illustrate use of the device by measuring ageing through replicative life span curves, following the dynamics of the cell cycle, and examining history-dependent behaviour in the general stress response.