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Open AccessArticle

A Combined AFM and Lateral Stretch Device Enables Microindentation Analyses of Living Cells at High Strains

1
Institute of Complex Systems: Biomechanics (ICS-7), Forschungszentrum Jülich, 52428 Jülich, Germany
2
Institute of Biology, Division of Cell and Developmental Biology, University of Leipzig, 04103 Leipzig, Germany
*
Author to whom correspondence should be addressed.
Methods Protoc. 2019, 2(2), 43; https://doi.org/10.3390/mps2020043
Received: 17 April 2019 / Revised: 13 May 2019 / Accepted: 21 May 2019 / Published: 24 May 2019
(This article belongs to the Special Issue Novel Approaches in Mechanobiology Research)
Mechanical characterization of living cells undergoing substantial external strain promises insights into material properties and functional principles of mechanically active tissues. However, due to the high strains that occur in physiological situations (up to 50%) and the complex structure of living cells, suitable experimental techniques are rare. In this study, we introduce a new system composed of an atomic force microscope (AFM), a cell stretching system based on elastomeric substrates, and light microscopy. With this system, we investigated the influence of mechanical stretch on monolayers of keratinocytes. In repeated indentations at the same location on one particular cell, we found significant stiffening at 25% and 50% strain amplitude. To study the contribution of intermediate filaments, we used a mutant keratinocyte cell line devoid of all keratins. For those cells, we found a softening in comparison to the wild type, which was even more pronounced at higher strain amplitudes. View Full-Text
Keywords: cell mechanics; cell stretching; atomic force microscopy; strain stiffening; cytokeratin network mechanics cell mechanics; cell stretching; atomic force microscopy; strain stiffening; cytokeratin network mechanics
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Ahrens, D.; Rubner, W.; Springer, R.; Hampe, N.; Gehlen, J.; Magin, T.M.; Hoffmann, B.; Merkel, R. A Combined AFM and Lateral Stretch Device Enables Microindentation Analyses of Living Cells at High Strains. Methods Protoc. 2019, 2, 43.

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