Table of Contents
ISRN Cell Biology
Volume 2012 (2012), Article ID 513546, 9 pages
http://dx.doi.org/10.5402/2012/513546
Research Article

An Approach to Visualize the Deformation of the Intermediate Filament Cytoskeleton in Response to Locally Applied Forces

Department of Physics and Department of Biology, University of Ottawa, MacDonald Hall,150 Louis Pasteur, Ottawa, ON, Canada K1N 6N5

Received 28 August 2011; Accepted 25 September 2011

Academic Editors: P. Lavia and C. C. Uphoff

Copyright © 2012 Jiashan Wang and Andrew E. Pelling. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The intermediate filament (IF) cytoskeleton plays an important role in integrating biomechanical pathways associated with the actin and microtubule cytoskeleton. Vimentin is a type III IF protein commonly found in fibroblast cells and plays a role in transmitting forces through the cytoskeleton. Employing simultaneous laser scanning confocal and atomic force microscopy (AFM), we developed a methodology to quantify the deformation of the GFP-vimentin-labeled IF cytoskeleton as a function of time in response to force application by the AFM. Over short times (seconds), IFs deformed rapidly and transmitted force throughout the entire cell in a highly complex and anisotropic fashion. After several minutes, mechanically induced displacements of IFs resemble basal movements. In well-adhered cells the deformation of IFs is highly anisotropic as they tend to deform away from the longitudinal axis of the cell. This study demonstrates that simultaneous AFM and LSCM can be employed to track the deformation and dissipation of force through the IF cytoskeleton.