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

Nat Protoc. 2015 Aug;10(8):1181-97. doi: 10.1038/nprot.2015.079. Epub 2015 Jul 9.


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 collecting data for thousands of single cells. Compared with other protocols, the present protocol is relatively easy to adopt and of higher throughput. The protocol can be widely used to control and monitor single-cell signaling dynamics in other signal transduction systems in microorganisms.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • DNA-Binding Proteins / metabolism*
  • High-Throughput Screening Assays*
  • Microfluidic Analytical Techniques*
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Signal Transduction*
  • Time-Lapse Imaging
  • Transcription Factors / metabolism*


  • DNA-Binding Proteins
  • MSN2 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors