Pump-free multi-well-based microfluidic system for high-throughput analysis of size-control relative genes in budding yeast.

Abstract

Time-lapse single cell imaging by microscopy can provide precise cell information such as the cell size, the cell cycle duration, protein localization and protein expression level. Usually, a microfluidic system is needed for these measurements in order to provide a constant culture environment and confine the cells so that they grow in a monolayer. However, complex connections are required between the channels inside the chip and the outside media, and a complex procedure is needed for loading of cells, thereby making this type of system unsuitable for application in high-throughput single cell scanning experiments. Here we provide a novel and easily operated pump-free multi-well-based microfluidic system which enables the high-throughput loading of many different budding yeast strains into monolayer growth conditions just by use of a multi-channel pipette. Wild type budding yeast (Saccharomyces cerevisiae) and 62 different budding yeast size control relative gene deletion strains were chosen for scanning. We obtained normalized statistical results for the mother cell doubling time, daughter cell doubling time, mother cell size and daughter cell size of different gene deletion strains relative to the corresponding parameters of the wild type cells. Meanwhile, we compared the typical cell morphology of different strains and analyzed the relationship between the cell genotype and phenotype. This method which can be easily used in a normal biology lab may help researchers who need to carry out the high-throughput scanning of cell morphology and growth.