QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

First published online 1 November 2005
doi: 10.1242/jcs.02653

This Article Figures Only Full Text Full Text (PDF) All Versions of this Article: jcs.02653v1 118/22/5315    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Related articles in JCS Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Franz, C. M. Articles by Müller, D. J. Search for Related Content PubMed PubMed Citation Articles by Franz, C. M. Articles by Müller, D. J.

Analyzing focal adhesion structure by atomic force microscopy

Center of Biotechnology, University of Technology Dresden, Tatzberg 49, 01307 Dresden, Germany

^* Author for correspondence (e-mail: mueller{at}biotec.tu-dresden.de)

Accepted 18 August 2005

Atomic force microscopy (AFM) can produce high-resolution topographic images of biological samples in physiologically relevant environments and is therefore well suited for the imaging of cellular surfaces. In this work we have investigated focal adhesion complexes by combined fluorescence microscopy and AFM. To generate high-resolution AFM topographs of focal adhesions, REF52 (rat embryo fibroblast) cells expressing YFP-paxillin as a marker for focal adhesions were de-roofed and paxillin-positive focal adhesions subsequently imaged by AFM. The improved resolution of the AFM topographs complemented the optical images and offered ultrastructural insight into the architecture of focal adhesions. Focal adhesions had a corrugated dorsal surface formed by microfilament bundles spaced 127±50 nm (mean±s.d.) apart and protruding 118±26 nm over the substratum. Within focal adhesions microfilaments were sometimes branched and arranged in horizontal layers separated by 10 to 20 nm. From the AFM topographs focal adhesion volumes could be estimated and were found to range from 0.05 to 0.50 µm³. Furthermore, the AFM topographs show that focal adhesion height increases towards the stress-fiber-associated end at an angle of about 3°. Finally, by correlating AFM height information with fluorescence intensities of YFP-paxillin and F-actin staining, we show that the localization of paxillin is restricted to the ventral half of focal adhesions, whereas F-actin-containing microfilaments reside predominantly in the membrane-distal half.

Key words: AFM, Focal adhesions, Paxillin

Related articles in JCS:

Taking the lid off focal adhesions

JCS 2005 118: 2104. [Full Text]  

This article has been cited by other articles:

				A. Nicolas, A. Besser, and S. A. Safran Dynamics of Cellular Focal Adhesions on Deformable Substrates: Consequences for Cell Force Microscopy Biophys. J., July 15, 2008; 95(2): 527 - 539. [Abstract] [Full Text] [PDF]

				J. Helenius, C.-P. Heisenberg, H. E. Gaub, and D. J. Muller Single-cell force spectroscopy J. Cell Sci., June 1, 2008; 121(11): 1785 - 1791. [Abstract] [Full Text] [PDF]

				C. Le Clainche and M.-F. Carlier Regulation of Actin Assembly Associated With Protrusion and Adhesion in Cell Migration Physiol Rev, April 1, 2008; 88(2): 489 - 513. [Abstract] [Full Text] [PDF]

				H. Shroff, C. G. Galbraith, J. A. Galbraith, H. White, J. Gillette, S. Olenych, M. W. Davidson, and E. Betzig Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes PNAS, December 18, 2007; 104(51): 20308 - 20313. [Abstract] [Full Text] [PDF]