Fluorescent-based quantitative measurements of signal transduction in single cells

Details

Serval ID
serval:BIB_CBA7FBEC6499
Type
A part of a book
Publication sub-type
Chapter: chapter ou part
Collection
Publications
Title
Fluorescent-based quantitative measurements of signal transduction in single cells
Title of the book
Design and Analysis of Biomolecular Circuits : engineering approaches to systems and synthetic biology
Author(s)
Pelet S., Peter M.
Publisher
Springer
Address of publication
New York
ISBN
978-1-4419-6765-7
Publication state
Published
Issued date
2011
Editor
Koeppl H., Densmore D., Setti G., di Bernardo M.
Chapter
17
Pages
369-393
Language
english
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.
Keywords
Cellular signaling, Fluorescent proteins, Microscopy, Flow cytometry, FRET, FCS
Create date
26/09/2012 11:39
Last modification date
20/08/2019 16:46
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