Multiplex imaging of Rho family GTPase activities in living cells

Désirée Spiering, Louis Hodgson

Research output: Chapter in Book/Report/Conference proceedingChapter

21 Scopus citations

Abstract

Here, we provide procedures for imaging the Rho GTPase biosensors in both single and multiplex acquisition modes. The multiplex approach enables the direct visualization of two biosensor readouts from a single living cell. Here, we take as an example a combination of the RhoA biosensor based on a CFP/YFP FRET modality and the Cdc42 biosensor based on organic dyes that change fluorescence as a function of the local solvent polarity. We list the required optical components as well as cellular manipulation techniques necessary to successfully image these two ratiometric biosensors in a single living cell.

Original languageEnglish (US)
Title of host publicationRho GTPases
Subtitle of host publicationMethods and Protocols
EditorsFrancisco Rivero
Pages215-234
Number of pages20
DOIs
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
Volume827
ISSN (Print)1064-3745

Keywords

  • Biosensors
  • Dynamics
  • FRET
  • Live-cell imaging
  • Multiplex
  • Rho GTPase

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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