Crane Matthew M, Clark Ivan B N, Bakker Elco, Smith Stewart, Swain Peter S
SynthSys - Center for Synthetic and Systems Biology and School of Biological Sciences, University of Edinburgh, Edinburgh, United Kingdom.
Scottish Microelectronics Center, University of Edinburgh, Edinburgh, United Kingdom.
PLoS One. 2014 Jun 20;9(6):e100042. doi: 10.1371/journal.pone.0100042. eCollection 2014.
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.
认识到细胞间变异性在细胞决策中的重要性以及对细胞过程随机建模的兴趣日益浓厚,导致在时变环境中对高密度、可重复的单细胞测量的需求增加。我们展示了ALCATRAS(长期培养和捕获系统),这是一种微流控装置,能够在明确且可控的环境中对多达1000个出芽酵母细胞进行定量监测。子细胞通过流体流动被移除,以避免拥挤,从而使实验能够运行超过60小时,并且细胞外培养基可以在数秒内反复更换。我们通过复制寿命曲线测量衰老、跟踪细胞周期动态以及检查一般应激反应中的历史依赖性行为来说明该装置的用途。