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Fluorescent-Based Quantitative Measurements of Signal Transduction in Single Cells

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Book cover Design and Analysis of Biomolecular Circuits

Abstract

Budding yeast (Saccharomyces cerevisiae) has been widely used as a model system to study fundamental biological processes. Genetic and biochemical approaches have allowed in the last decades to uncover the key components involved in many signaling pathways. Generally, most techniques measure the average behavior of a population of cells, and thus miss important cell-to-cell variations. With the recent progress in fluorescent proteins, new avenues have been opened to quantitatively study the dynamics of signaling in single living cells. In this chapter, we describe several techniques based on fluorescence measurements to quantify the activation of biological pathways. Flow cytometry allows for rapid quantification of the total fluorescence of a large number of single cells. In contrast, microscopy offers a lower throughput but allows to follow with a high temporal resolution the localization of proteins at sub-cellular resolution. Finally, advanced functional imaging techniques such as FRET and FCS offer the possibility to directly visualize the formation of protein complexes or to quantify the activity of proteins in vivo. Together these techniques present powerful new approaches to study cellular signaling and will greatly increase our understanding of the regulation of signaling networks in budding yeast and beyond.

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Acknowledgements

We would like to thank Reinhard Dechant for critical reading of the manuscript. Work in the laboratory of M.P. is supported by Unicellsys, SPMD, the SystemsX.ch initiative (YeastX and LiverX projects), the ETHZ and the Swiss National Science Foundation (SNF).

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Authors and Affiliations

  1. Department of Biology, Institute of Biochemistry, Schaffmattstr 18, CH 8093-Zürich, ETH Zürich, Switzerland

    Serge Pelet

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  1. Serge Pelet
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  2. Matthias Peter
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Correspondence to Serge Pelet .

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Editors and Affiliations

  1. , Automatic Control Laboratory, ETH Zurich, Physikstrasse 3, Zurich, 8092, Switzerland

    Heinz Koeppl

  2. ARCES, Università di Bologna, Via Toffano 2/2, Bologna, 40125, Italy

    Gianluca Setti

  3. Università di Napoli Federico II, Via Claudio 21, Napoli, 80125, Italy

    Mario di Bernardo

  4. , Department of Electrical and Computer En, Boston University, Saint Mary's St. 8, Boston, 02446, Massachusetts, USA

    Douglas Densmore

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Pelet, S., Peter, M. (2011). Fluorescent-Based Quantitative Measurements of Signal Transduction in Single Cells. In: Koeppl, H., Setti, G., di Bernardo, M., Densmore, D. (eds) Design and Analysis of Biomolecular Circuits. Springer, New York, NY. https://doi.org/10.1007/978-1-4419-6766-4_17

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  • DOI: https://doi.org/10.1007/978-1-4419-6766-4_17

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