High-throughput single-cell analysis for the proteomic dynamics study of the yeast osmotic stress response.
High-throughput single-cell analysis for the proteomic dynamics study of the yeast osmotic stress response.
复制标题
用于酵母渗透应激反应蛋白质组动力学研究的高通量单细胞分析
DOI：
10.1038/srep42200
复制
发表时间：
2017-02-09
影响因子：
4.6
通讯作者：
Luo C
中科院分区：
文献类型：
--
作者：
Zhang R;Yuan H;Wang S;Ouyang Q;Chen Y;Hao N;Luo C
在细胞生物学研究中，电动荧光显微镜结合高通量微流控芯片是获得不同生物过程信息的有力手段。通常，为了在不同的环境下观察不同的菌株，高通量微流控芯片需要复杂的准备工作。在这项研究中，我们设计了一种新的，易于操作的高通量微流控系统，观察96个不同的GFP标记的酵母菌株在一个可切换的培养条件或24个不同的GFP标记的酵母菌株在四个平行的可切换的培养条件。多移液管是芯片中高通量细胞图案化所需的唯一附加设备。仅需8个连接即可控制96个条件。使用这些装置，酵母应激反应途径的蛋白质组动力学进行了仔细研究的基础上单细胞数据。提出了一种新的方法来表征蛋白质组动力学使用单细胞的数据，并与以前的方法相比，新技术应该是有用的研究底层控制网络。我们的方法提供了一个简单和系统的方法来研究信号通路在单细胞水平。
Motorized fluorescence microscopy combined with high-throughput microfluidic chips is a powerful method to obtain information about different biological processes in cell biology studies. Generally, to observe different strains under different environments, high-throughput microfluidic chips require complex preparatory work. In this study, we designed a novel and easily operated high-throughput microfluidic system to observe 96 different GFP-tagged yeast strains in one switchable culture condition or 24 different GFP-tagged yeast strains in four parallel switchable culture conditions. A multi-pipette is the only additional equipment required for high-throughput patterning of cells in the chip. Only eight connections are needed to control 96 conditions. Using these devices, the proteomic dynamics of the yeast stress response pathway were carefully studied based on single-cell data. A new method to characterize the proteomic dynamics using a single cell’s data is proposed and compared to previous methods, and the new technique should be useful for studying underlying control networks. Our method provides an easy and systematic way to study signaling pathways at the single-cell level.