High-throughput microfluidics to control and measure signaling dynamics in single yeast cells.

ABSTRACT: Microfluidics coupled to quantitative time-lapse fluorescence microscopy is transforming our ability to control, measure and understand signaling dynamics in single living cells. Here we describe a pipeline that incorporates multiplexed microfluidic cell culture, automated programmable fluid handling for cell perturbation, quantitative time-lapse microscopy and computational analysis of time-lapse movies. We illustrate how this setup can be used to control the nuclear localization of the budding yeast transcription factor Msn2. By using this protocol, we generate oscillations of Msn2 localization and measure the dynamic gene expression response of individual genes in single cells. The protocol allows a single researcher to perform up to 20 different experiments in a single day, while colle

SUBMITTER: Hansen AS 

PROVIDER: S-EPMC4593625 | biostudies-literature | 2015 Aug

REPOSITORIES: biostudies-literature