July 2010 Nature publishes articles on the use of high-throughput microfluidics to study the operation of single-cell biological systems
Introduction
In a dynamic environment, cells work through the interaction of genetic pathways. Mammalian immune responses are a striking example of the synergy of different types of cells. Cell-to-cell communication is primarily mediated by the formation of temporal and spatial concentration gradients of signal molecules, which requires cells to respond to a wide range of signal intensities. The article microfluidic cell culture using high-throughput (high-throughput microfluidic cell culture) and fluorescence microscopy, quantitative analysis of gene expression studies and mathematical models in the end means to assess how a single mammalian cell signaling molecules to different concentrations of TNF - α produces a corresponding response. The value of high-throughput gene quantification at the single-cell level to study how biological systems work is highly recognized.
Letter
Nature 466, 267-271 (8 July 2010) | doi:10.1038/nature09145; Received 29 December 2009; Accepted 28 April 2010; Published online 27 June 2010
Single-cell NF- κ B dynamics reveal digital activation and analogue information processing Sava ş Tay1,2,4, Jacob J. Hughey1,4, Timothy K. Lee1, Tomasz Lipniacki3, Stephen R. Quake1,2 & Markus W. Covert1
1. Â Department of Bioengineering, Stanford University, Stanford, California 94305, USA
2. Â Howard Hughes Medical Institute, Stanford, California 94305, USA
3. Â Institute of Fundamental Technological Research, Warsaw 02-106, Poland
4. Â These authors contributed equally to this work.
Abstract
Cells operating in dynamic environments using extraordinary communication capabilities that emerge from the interactions of genetic circuitry. The mammalian immune response is a striking example of the coordination of different cell types 1. Cell-to-cell communication is primarily mediated by signalling molecules that form spatiotemporal concentration Gradients, requiring cells to respond to a wide range of signal intensities2. Here we use high-throughput microfluidic cell culture3 and fluorescence microscopy, quantitative gene expression analysis and mathematical modelling to investigate how single mammalian cells respond to different concentrations of the signaling molecule tumour-necrosis Factor (TNF) -α , and relay information to the gene expression programs by means of the transcription factor nuclear factor (NF)- κ B. We measured NF- κ B activity in thousands of live cells under TNF- α doses covering four orders Of magnitude. We find, in contrast to population-level studies with Bulk assays2, that the activation is heterogeneous and is a digital process at the single-cell level with the cells responding at lower doses. Cells also encode a subtle set of analogue parameters to modulate the outcome; these parameters include NF- κ B peak intensity We developed a stochastic mathematical model that reproduces both the digital and analogue dynamics as well as most gene expression profiles at all measured conditions, constituting a broadly applicable model for TNF- α -induced NF- κ B signalling In various types of cells. These results highlight the value of high-throughput quantitative measurements with single-cell resolution in understanding how biological systems operate.
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