Literature DB >> 22404943

In vivo imaging of the actin polymerization state with two-photon fluorescence anisotropy.

Harshad D Vishwasrao1, Pierre Trifilieff, Eric R Kandel.   

Abstract

Using two-photon fluorescence anisotropy imaging of actin-GFP, we have developed a method for imaging the actin polymerization state that is applicable to a broad range of experimental systems extending from fixed cells to live animals. The incorporation of expressed actin-GFP monomers into endogenous actin polymers enables energy migration FRET (emFRET, or homoFRET) between neighboring actin-GFPs. This energy migration reduces the normally high polarization of the GFP fluorescence. We derive a simple relationship between the actin-GFP fluorescence polarization anisotropy and the actin polymer fraction, thereby enabling a robust means of imaging the actin polymerization state with high spatiotemporal resolution and providing what to the best of our knowledge are the first direct images of the actin polymerization state in live, adult brain tissue and live, intact Drosophila larvae.
Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.

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Year:  2012        PMID： 22404943      PMCID： PMC3296026          DOI： 10.1016/j.bpj.2012.01.031

Source DB:  PubMed          Journal:  Biophys J        ISSN： 0006-3495            Impact factor:   4.033


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