Multiplex imaging of Rho family GTPase activities in living cells

Désirée Spiering, Louis Hodgson

Research output: Chapter in Book/Report/Conference proceedingChapter

18 Citations (Scopus)

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 publicationMethods in Molecular Biology
Pages215-234
Number of pages20
Volume827
DOIs
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
Volume827
ISSN (Print)10643745

Fingerprint

rho GTP-Binding Proteins
Biosensing Techniques
Optical Devices
Coloring Agents
Fluorescence

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Spiering, D., & Hodgson, L. (2012). Multiplex imaging of Rho family GTPase activities in living cells. In Methods in Molecular Biology (Vol. 827, pp. 215-234). (Methods in Molecular Biology; Vol. 827). https://doi.org/10.1007/978-1-61779-442-1_15

Multiplex imaging of Rho family GTPase activities in living cells. / Spiering, Désirée; Hodgson, Louis.

Methods in Molecular Biology. Vol. 827 2012. p. 215-234 (Methods in Molecular Biology; Vol. 827).

Research output: Chapter in Book/Report/Conference proceedingChapter

Spiering, D & Hodgson, L 2012, Multiplex imaging of Rho family GTPase activities in living cells. in Methods in Molecular Biology. vol. 827, Methods in Molecular Biology, vol. 827, pp. 215-234. https://doi.org/10.1007/978-1-61779-442-1_15
Spiering D, Hodgson L. Multiplex imaging of Rho family GTPase activities in living cells. In Methods in Molecular Biology. Vol. 827. 2012. p. 215-234. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-61779-442-1_15
Spiering, Désirée ; Hodgson, Louis. / Multiplex imaging of Rho family GTPase activities in living cells. Methods in Molecular Biology. Vol. 827 2012. pp. 215-234 (Methods in Molecular Biology).
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