Atomic Force Microscopy and High-Content Analysis: Two Innovative Technologies for Dissecting the Relationship Between Epithelial–Mesenchymal Transition-Related Morphological and Structural Alterations and Cell Mechanical Properties

  • Stephen T. BuckleyEmail author
  • Anthony M. Davies
  • Carsten Ehrhardt
Part of the Methods in Molecular Biology book series (MIMB, volume 784)


Epithelial–mesenchymal transition (EMT) is a complex series of cellular reprogramming events culminating in striking alterations in morphology towards an invasive mesenchymal phenotype. Increasingly, evidence suggests that EMT exerts a pivotal role in pathophysiological situations including fibrosis and cancer. Core to these dynamical changes in cellular polarity and plasticity is discrete modifications in cytoskeletal structure. In particular, newly established actin-stress fibres supplant a preceding system of highly organised cortical actin. Although cumulative studies have contributed to elucidation of the detailed signalling pathways that underpin this elaborate molecular process, there remains a deficiency regarding its precise contribution to cellular biomechanics. The advent of atomic force microscopy (AFM) and high-content analysis (HCA) provides two innovative technologies for dissecting the relationship between EMT-related morphological and structural alterations and cell mechanical properties. AFM permits acquisition of high resolution topographical images and detailed analysis of cellular viscoelasticity while HCA facilitates a comprehensive and perspicacious assessment of morphological changes. In combination, they offer the possibility of novel insights into the dynamic traits of transitioning cells. Herein, a detailed protocol describing AFM and HCA techniques for evaluation of transforming growth factor-β1-induced EMT of alveolar epithelial cells is provided.

Key words

Cytoskeleton Epithelial–mesenchymal transition Atomic force microscopy High-content analysis microscopy 



STB is funded by an IRCSET Government of Ireland Postgraduate Scholarship in Science, Engineering and Technology. This work has been funded in part by a Strategic Research Cluster grant (07/SRC/B1154) under the National Development Plan co-funded by EU Structural Funds and Science Foundation Ireland.


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Stephen T. Buckley
    • 1
    Email author
  • Anthony M. Davies
    • 2
  • Carsten Ehrhardt
    • 1
  1. 1.School of Pharmacy & Pharmaceutical SciencesTrinity College DublinDublinIreland
  2. 2.Department of Clinical MedicineTrinity College DublinDublinIreland

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