@inbook{d2a44a3aa14c41508a4beec62846bd5b,
title = "Using fluorescence resonance energy transfer-based biosensors to probe Rho GTPase activation during phagocytosis",
abstract = "The p21-family members of Rho GTPases are important for the control of actin cytoskeleton dynamics, and are critical regulators of phagocytosis. The three-dimensional structure of phagosomes and the highly compartmentalized nature of the signaling mechanisms during phagocytosis require high-resolution imaging using ratiometric biosensors to decipher Rho GTPase activities regulating phagosome formation and function. Here we describe methods for the expression and ratiometric imaging of FRET-based Rho GTPase biosensors in macrophages during phagocytosis. As an example, we show Cdc42 activity at the phagosome over Z-serial planes. In addition, we demonstrate the usage of a new, fast, and user-friendly deconvolution package that delivers significant improvements in the attainable details of Rho GTPase activity in phagosome structures.",
keywords = "Biosensors, Deconvolution, FRET, Macrophages, Phagosome, Ratiometric imaging, Z-stack",
author = "Veronika Miskolci and Louis Hodgson and Dianne Cox",
note = "Publisher Copyright: {\textcopyright} Springer Science+Business Media New York 2017.",
year = "2017",
doi = "10.1007/978-1-4939-6581-6_9",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "125--143",
booktitle = "Methods in Molecular Biology",
}