Measuring Cell–Cell Tugging Forces Using Bowtie-Patterned mPADs (Microarray Post Detectors)

  • Daniel M. Cohen
  • Mike T. Yang
  • Christopher S. Chen
Part of the Methods in Molecular Biology book series (MIMB, volume 1066)


Cells generate traction forces upon adhesion to the extracellular matrix as well as to neighboring cells. These forces are important for the growth and maintenance of adhesion structures such as focal adhesions and adherens junctions, and may play roles in tissue development. Here, we describe a method for measuring the tugging force transmitted across the cell–cell junction between two paired cells.

Key words

Cell–cell tugging force Cell–cell junctions Micropatterning mPADs Traction force 



This work was supported by NIH grants HL73305 and through the RESBIO program (Integrated Technology Resource for Polymeric Biomaterials, NIH grant EB001046).


  1. 1.
    Lee J, Leonard M, Oliver T (1994) Traction forces generated by locomoting keratocytes. J Cell Biol 127:1957–1964PubMedCrossRefGoogle Scholar
  2. 2.
    Pelham RJ, Wang Y (1999) High resolution detection of mechanical forces exerted by locomoting fibroblasts on the substrate. Mol Biol Cell 10:935–945PubMedCrossRefGoogle Scholar
  3. 3.
    Balaban NQ, Schwarz US, Riveline D et al (2001) Force and focal adhesion assembly: a close relationship studied using elastic micropatterned substrates. Nat Cell Biol 3:466–472PubMedCrossRefGoogle Scholar
  4. 4.
    Tan JL, Tien J, Pirone DM et al (2003) Cells lying on a bed of microneedles: an approach to isolate mechanical force. Proc Natl Acad Sci USA 100:1484–1489Google Scholar
  5. 5.
    Perret E, Leung A, Feracci H et al (2004) Trans-bonded pairs of E-cadherin exhibit a remarkable hierarchy of mechanical strengths. Proc Natl Acad Sci USA 101:16472–16477Google Scholar
  6. 6.
    Baumgartner W, Hinterdorfer P, Ness W et al (2009) Cadherin interaction probed by atomic force microscopy. Proc Natl Acad Sci USA 97:4005–4010Google Scholar
  7. 7.
    Panorchan P, Thompson MS, Davis KJ et al (2006) Single-molecule analysis of cadherin-mediated cell-cell adhesion. J Cell Sci 119:66–74PubMedCrossRefGoogle Scholar
  8. 8.
    Potard US, Butler JP, Wang N (1997) Cytoskeletal mechanics in confluent epithelial cells probed through integrins and E-cadherins. Am J Physiol 272:C1654–1663PubMedGoogle Scholar
  9. 9.
    Ko KS, Arora PD, McCulloch CA (2001) Cadherins mediate intercellular mechanical signaling in fibroblasts by activation of stretch-sensitive calcium-permeable channels. J Biol Chem 276:35967–35977PubMedCrossRefGoogle Scholar
  10. 10.
    Felsenfeld DP, Choquet D, Sheetz MP (1996) Ligand binding regulates the directed movement of beta1 integrins on fibroblasts. Nature 383:438–440PubMedCrossRefGoogle Scholar
  11. 11.
    Galbraith CG, Yamada KM, Sheetz MP (2002) The relationship between force and focal complex development. J Cell Biol 159:695–705PubMedCrossRefGoogle Scholar
  12. 12.
    Ohashi T, Kiehart DP, Erickson HP (2002) Dual labeling of the fibronectin matrix and actin cytoskeleton with green fluorescent protein variants. J Cell Sci 115:1221–1229PubMedGoogle Scholar
  13. 13.
    Smith ML, Gourdon D, Little WC et al (2007) Force-induced unfolding of fibronectin in the extracellular matrix of living cells. PLoS Biol 5:e268PubMedCrossRefGoogle Scholar
  14. 14.
    Kong HJ, Polte TR, Alsberg E et al (2005) FRET measurements of cell-traction forces and nano-scale clustering of adhesion ligands varied by substrate stiffness. Proc Natl Acad Sci USA 102:4300–4305Google Scholar
  15. 15.
    Grashoff C, Hoffman BD, Brenner MD et al (2010) Measuring mechanical tension across vinculin reveals regulation of focal adhesion dynamics. Nature 466:263–266PubMedCrossRefGoogle Scholar
  16. 16.
    Liu Z, Tan JL, Cohen DM et al (2010) Mechanical tugging force regulates the size of cell-cell junctions. Proc Natl Acad Sci USA 107:9944–9949PubMedCrossRefGoogle Scholar
  17. 17.
    Nelson CM, Liu WF, Chen CS (2007) Manipulation of cell-cell adhesion using bowtie-shaped microwells. Methods Mol Biol 370:1–10PubMedCrossRefGoogle Scholar
  18. 18.
    Maruthamuthu V, Sabass B, Schwarz US et al (2011) Cell-ECM traction force modulates endogenous tension at cell-cell contacts. Proc Natl Acad Sci USA 108:4708–4713PubMedCrossRefGoogle Scholar
  19. 19.
    Miyake Y, Inoue N, Nishimura K et al (2006) Actomyosin tension is required for correct recruitment of adherens junction components and zonula occludens formation. Exp Cell Res 312:1637–1650PubMedCrossRefGoogle Scholar
  20. 20.
    Shewan AM, Maddugoda M, Kraemer A et al (2005) Myosin 2 is a key Rho kinase target necessary for the local concentration of E-cadherin at cell-cell contacts. Mol Biol Cell 16:4531–4542PubMedCrossRefGoogle Scholar
  21. 21.
    Huveneers S, Oldenburg J, Spanjaard E et al (2012) Vinculin associates with endothelial VE-cadherin junctions to control force-dependent remodeling. J Cell Biol 196:641–652PubMedCrossRefGoogle Scholar
  22. 22.
    Yonemura S, Wada Y, Watanabe T et al (2010) alpha-Catenin as a tension transducer that induces adherens junction development. Nat Cell Biol 12:533–542PubMedCrossRefGoogle Scholar
  23. 23.
    Yang MT, Reich DH, Chen CS (2011) Measurement and analysis of traction force dynamics in response to vasoactive agonists. Integr Biol (Camb) 3:663–674CrossRefGoogle Scholar
  24. 24.
    Yang MT, Fu J, Wang YK et al (2011) Assaying stem cell mechanobiology on microfabricated elastomeric substrates with geometrically modulated rigidity. Nat Protoc 6:187–213PubMedCrossRefGoogle Scholar
  25. 25.
    Sniadecki NJ, Chen CS (2007) Microfabricated silicone elastomeric post arrays for measuring traction forces of adherent cells. Methods Cell Biol 83:313–328PubMedCrossRefGoogle Scholar
  26. 26.
    Desai RA, Yang MT, Sniadecki NJ et al (2007) Microfabricated post-array-detectors (mPADs): an approach to isolate mechanical forces. J Vis Exp 8:e311Google Scholar
  27. 27.
    Shen K, Qi J, Kam LC (2008) Microcontact printing of proteins for cell biology. J Vis Exp 22:e1065Google Scholar
  28. 28.
    Shaw SK, Bamba PS, Perkins BN et al (2001) Real-time imaging of vascular endothelial-cadherin during leukocyte transmigration across endothelium. J Immunol 167:2323–2330PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Daniel M. Cohen
    • 1
  • Mike T. Yang
    • 1
  • Christopher S. Chen
    • 1
  1. 1.Department of BioengineeringUniversity of PennsylvaniaPhiladelphiaUSA

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