Abstract
The inability to measure mechanical forces within cells has been limiting our understanding of how mechanical information is processed on the molecular level. In this chapter, we describe a method that allows the analysis of force propagation across distinct proteins within living cells using Förster resonance energy transfer (FRET)-based biosensors.
Katharina Austen, Carleen Kluger, and Andrea Freikamp have contributed equally to this chapter.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Hoffman BD, Grashoff C, Schwartz MA (2011) Dynamic molecular processes mediate cellular mechanotransduction. Nature 475:316–323
Bao G, Suresh S (2003) Cell and molecular mechanics of biological materials. Nat Mater 2:715–725
Wang JH, Li B (2009) Application of cell traction force microscopy for cell biology research. Methods Mol Biol 586:301–313
Rowat AC (2009) Physical properties of the nucleus studied by micropipette aspiration. Methods Mol Biol 464:3–12
Mackay JL, Kumar S (2013) Measuring the elastic properties of living cells with atomic force microscopy indentation. Methods Mol Biol 931:313–329
Kasza KE, Vader D, Koster S et al (2011) Magnetic twisting cytometry. Cold Spring Harb Protoc 2011:pdp prot5599
Grashoff C, Hoffman BD, Brenner MD et al (2010) Measuring mechanical tension across vinculin reveals regulation of focal adhesion dynamics. Nature 466:263–266
Doyle AD, Yamada KM (2010) Cell biology: Sensing tension. Nature 466:192–193
Jares-Erijman EA, Jovin TM (2006) Imaging molecular interactions in living cells by FRET microscopy. Curr Opin Chem Biol 10:409–416
Jares-Erijman EA, Jovin TM (2003) FRET imaging. Nat Biotechnol 21:1387–1395
Morton PE, Parsons M (2011) Measuring FRET using time-resolved FLIM. Methods Mol Biol 769:403–413
Becker W (2012) Fluorescence lifetime imaging – techniques and applications. J Microsc 247:119–136
Nelson MD, Fitch DH (2011) Overlap extension PCR: an efficient method for transgene construction. Methods Mol Biol 772:459–470
Heckman KL, Pease LR (2007) Gene splicing and mutagenesis by PCR-driven overlap extension. Nat Protoc 2:924–932
Swift S, Lorens J, Achacoso P, et al (2001) Rapid production of retroviruses for efficient gene delivery to mammalian cells using 293T cell-based systems. In: John E. Coligan et al. (eds.) Current protocols in immunology. Chapter 10, Unit 10 17C
Digman MA, Caiolfa VR, Zamai M et al (2008) The phasor approach to fluorescence lifetime imaging analysis. Biophys J 94:L14–16
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Austen, K., Kluger, C., Freikamp, A., Chrostek-Grashoff, A., Grashoff, C. (2013). Generation and Analysis of Biosensors to Measure Mechanical Forces Within Cells. In: Baudino, T. (eds) Cell-Cell Interactions. Methods in Molecular Biology, vol 1066. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-604-7_15
Download citation
DOI: https://doi.org/10.1007/978-1-62703-604-7_15
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-603-0
Online ISBN: 978-1-62703-604-7
eBook Packages: Springer Protocols