Stratz Simone, Verboket Pascal Emilio, Hasler Karina, Dittrich Petra Stephanie
ETH Zurich, Department of Chemistry and Applied Biosciences, Zurich, Switzerland.
ETH Zurich, Department of Biosystems Science and Engineering, Basel, Switzerland.
Electrophoresis. 2018 Feb;39(3):540-547. doi: 10.1002/elps.201700253. Epub 2017 Oct 4.
Here, we present a multifunctional microfluidic device whose integrative design enables to combine cell culture studies and quantitative single cell biomolecule analysis. The platform consists of 32 analysis units providing two key features; first, a micrometer-sized trap for hydrodynamic capture of a single Saccharomyces cerevisiae (S. cerevisiae) yeast cell; second, a convenient double-valve configuration surrounding the trap. Actuating of the outer valve with integrated opening results in a partial isolation in a volume of 11.8 nL, i.e. the cell surrounding fluid can be exchanged diffusion-based without causing shear stress or cell loss. Actuation of the inner ring-shaped valve isolates the trapped cell completely in a small analysis volume of 230 pL. The device was used to determine the growth rate of yeast cells (S. cerevisiae) under under optimum and oxidative stress conditions. In addition, we successfully quantified the cofactor beta-nicotinamide adenine dinucleotide phosphate (NAD(P)H) in single and few cells exposed to the different microenvironments. In conclusion, the microdevice enables to analyze the influence of an external stress factor on the cellular fitness in a fast and more comprehensive way as cell growth and intracellular biomolecule levels can be investigated.
在此，我们展示了一种多功能微流控装置，其一体化设计能够将细胞培养研究与定量单细胞生物分子分析相结合。该平台由32个分析单元组成，具有两个关键特性：第一，一个微米级的阱，用于通过流体动力学捕获单个酿酒酵母（S. cerevisiae）酵母细胞；第二，围绕阱的便捷双阀配置。激活带有集成开口的外阀会导致在11.8 nL的体积内实现部分隔离，即细胞周围的流体可以基于扩散进行交换，而不会引起剪切应力或细胞损失。激活内环阀可将捕获的细胞完全隔离在230 pL的小分析体积中。该装置用于确定酵母细胞（S. cerevisiae）在最佳条件和氧化应激条件下的生长速率。此外，我们成功地对暴露于不同微环境的单个细胞和少量细胞中的辅因子β - 烟酰胺腺嘌呤二核苷酸磷酸（NAD(P)H）进行了定量。总之，该微装置能够以快速且更全面的方式分析外部应激因素对细胞适应性的影响，因为可以研究细胞生长和细胞内生物分子水平。
