Microbial single-cell growth response at defined carbon limiting conditions

Growth is one of the most fundamental characteristics of life, but detailed knowledge regarding growth at nutrient limiting conditions remains scarce. In recent years progress in microfluidic single-cell analysis and cultivation techniques has given insights into many fundamental growth characteristics such as growth homeostasis, aging and cell division of microbial cells. Using microfluidic single-cell cultivation technologies we examined how single-cell growth at defined carbon conditions, ranging from strongly limiting conditions (0.01 mmol L−1) to a carbon surplus (100 mmol L−1), influenced cell-to-cell variability. The experiments showed robust growth of populations at intermediate concentrations and cell-to-cell variability was higher at low and high carbon concentrations, among an isogenic population. Single-cell growth at extremely limiting conditions led not only to significant variability of division times, but also to an increased number of cells that did not pursue growth. Overall, the results demonstrate that cellular behaviour shows robust, Monod-like growth, with significant cell-to-cell heterogeneity at extreme limiting conditions, resembling natural habitats. Due to this significant influence of the environment on cellular physiology, more carefulness needs to be given future microfluidic single-cell experiments. Consequently, our results lay the foundation for the re-interpretation and design of workflows for future experiments aiming at an improved understanding of cell growth mechanisms