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Rho GTPases pp 215-234 | Cite as

Multiplex Imaging of Rho Family GTPase Activities in Living Cells

  • Désirée Spiering
  • Louis Hodgson
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 827)

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.

Key words

Rho GTPase Dynamics Live-cell imaging FRET Biosensors Multiplex 

Notes

Acknowledgments

This work was funded by GM093121 (D.S. and L.H.) and “Sinsheimer Foundation Young Investigator Award” (L.H.).

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Anatomy and Structural Biology and Gruss-Lipper Biophotonics CenterAlbert Einstein College of Medicine of Yeshiva UniversityBronxUSA

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