Fluctuation analysis of activity biosensor images for the study of information flow in signaling pathways

Marco Vilela, Nadia Halidi, Sebastien Besson, Hunter Elliott, Klaus Hahn, Jessica Tytell, Gaudenz Danuser

Research output: Chapter in Book/Report/Conference proceedingChapter

19 Scopus citations


Comprehensive understanding of cellular signal transduction requires accurate measurement of the information flow in molecular pathways. In the past, information flow has been inferred primarily from genetic or protein-protein interactions. Although useful for overall signaling, these approaches are limited in that they typically average over populations of cells. Single-cell data of signaling states are emerging, but these data are usually snapshots of a particular time point or limited to averaging over a whole cell. However, many signaling pathways are activated only transiently in specific subcellular regions. Protein activity biosensors allow measurement of the spatiotemporal activation of signaling molecules in living cells. These data contain highly complex, dynamic information that can be parsed out in time and space and compared with other signaling events as well as changes in cell structure and morphology. We describe in this chapter the use of computational tools to correct, extract, and process information from time-lapse images of biosensors. These computational tools allow one to explore the biosensor signals in a multiplexed approach in order to reconstruct the sequence of signaling events and consequently the topology of the underlying pathway. The extraction of this information, dynamics and topology, provides insight into how the inputs of a signaling network are translated into its biochemical or mechanical outputs.

Original languageEnglish (US)
Title of host publicationFluorescence Fluctuation Spectroscopy (FFS), Part B
PublisherAcademic Press Inc.
Number of pages24
ISBN (Print)9780124055391
StatePublished - 2013

Publication series

NameMethods in Enzymology
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988


  • Biosensor
  • Correlation
  • FRET
  • Image analysis
  • Information flow
  • Signal transduction

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology


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