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

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

SUBMITTER: Vishwasrao HD 

PROVIDER: S-EPMC3296026 | biostudies-literature | 2012 Mar

REPOSITORIES: biostudies-literature