Feeling for Filaments: Quantification of the Cortical Actin Web in Live Vascular Endothelium.

ABSTRACT: Contact-mode atomic force microscopy (AFM) has been shown to reveal cortical actin structures. Using live endothelial cells, we visualized cortical actin dynamics simultaneously by AFM and confocal fluorescence microscopy. We present a method that quantifies dynamic changes in the mechanical ultrastructure of the cortical actin web. We argue that the commonly used, so-called error signal imaging in AFM allows a qualitative, but not quantitative, analysis of cortical actin dynamics. The approach we used comprises fast force-curve-based topography imaging and subsequent image processing that enhances local height differences. Dynamic changes in the organization of the cytoskeleton network can be observed and quantified by surface roughness calculations and automated morphometrics. Upon treat

SUBMITTER: Kronlage C 

PROVIDER: S-EPMC4547164 | biostudies-literature | 2015 Aug

REPOSITORIES: biostudies-literature