Abstract
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.
Highlights
Motorized fluorescence microscopy combined with high-throughput microfluidic chips is a powerful method to obtain information about different biological processes in cell biology studies
Increasing numbers of scientists have realized that high-throughput studies of proteomic dynamics of single cells rather than steady behavior experiments are needed to determine the functions of biological networks[1,2]
The 96 injection wells of different cell strains are arrayed as four lines on two sides of the chip separated by 3 mm
Summary
Motorized fluorescence microscopy combined with high-throughput microfluidic chips is a powerful method to obtain information about different biological processes in cell biology studies. 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. In the past decade, motorized fluorescence microscopy has become a powerful tool for chemical analysis and quantitative biological measurements because it can obtain accurate real-time images of cell morphology[12], such as the expression level and location of proteins, cell shape, and other dynamic changes In these studies, microfluidic devices are often used to control the cell location, cell growth direction and micro environment to obtain high-quality cell images[13,14,15,16,17]. If Taylor’s device was modified to study the dynamics of proteins in one signaling pathway (usually one signaling pathway contains tens to hundreds of proteins), it would be too complicated because it would require hundreds of pipelines in one chip
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