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Quantifying cell-generated mechanical forces within living embryonic tissues

A Corrigendum to this article was published on 27 February 2014

This article has been updated


Cell-generated mechanical forces play a critical role during tissue morphogenesis and organ formation in the embryo. Little is known about how these forces shape embryonic organs, mainly because it has not been possible to measure cellular forces within developing three-dimensional (3D) tissues in vivo. We present a method to quantify cell-generated mechanical stresses exerted locally within living embryonic tissues, using fluorescent, cell-sized oil microdroplets with defined mechanical properties and coated with adhesion receptor ligands. After a droplet is introduced between cells in a tissue, local stresses are determined from droplet shape deformations, measured using fluorescence microscopy and computerized image analysis. Using this method, we quantified the anisotropic stresses generated by mammary epithelial cells cultured within 3D aggregates, and we confirmed that these stresses (3.4 nN μm−2) are dependent on myosin II activity and are more than twofold larger than stresses generated by cells of embryonic tooth mesenchyme, either within cultured aggregates or in developing whole mouse mandibles.

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Figure 1: Oil microdroplets as force transducers.
Figure 2: Measure of cell-generated mechanical stresses in epithelial and mesenchymal cell aggregates.
Figure 3: Ensemble statistics of droplet deformations in cell aggregates.
Figure 4: Measurement of cell-generated mechanical stresses in living mandibles.
Figure 5: Statistics of droplet deformations in living mandibles.

Change history

  • 05 February 2014

    In the version of this article initially published, the current affiliation of author Ralph Sperling was not included. His current affiliation is the Fraunhofer ICT-IMM, Mainz, Germany. The error has been corrected in the HTML and PDF versions of the article.


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We thank the SysCODE consortium for postdoctoral financial support for O.C. and for interesting discussions with several of its members. We thank C. Jorcyk (Boise State University) for providing premalignant mammary epithelial M28 cells and B. Ristenpart for the Matlab code used to analyze data obtained with the pendant drop method. O.C. thanks all members of the Ingber lab for their help and support, J. Gros for help with imaging, and F. Aguet for help with SteerableJ plugins. R.A.S. gratefully acknowledges funding from the German Research Foundation (Sp 1282/1-1). This work was supported by US National Institutes of Health grant RL1 DE019023-01 (to D.E.I.), the Wyss Institute for Biologically Inspired Engineering at Harvard University, the MacArthur Foundation and the Harvard NSF-MRSEC (L.M.).

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Authors and Affiliations



D.E.I., O.C. and L.M. defined the project; O.C. conceived of the droplets as force transducers; O.C. and D.E.I. designed the technique; T.M. and D.O. provided dissected mouse mandibles; O.C. and S.H. microinjected droplets into mouse mandibles; O.C., R.A.S. and D.A.W. designed and synthesized new fluorocarbon-hydrocarbon block copolymers; O.C. and A.G.B. did the initial tests of the technique using cell aggregates; O.C. performed force measurements in cell-drop aggregates and living mouse mandibles; O.C. performed confocal measurements; O.C. analyzed the data; D.O. and R.M. provided transgenic mice; and O.C., L.M. and D.E.I. wrote the paper.

Corresponding authors

Correspondence to Otger Campàs or Donald E Ingber.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1 and 2 and Supplementary Notes 1–3 (PDF 4059 kb)

Effect of myosin II inhibition on droplet deformations

Time-lapse showing the effect of myosin II inhibition on droplet deformations. Myosin II was inhibited using blebbistatin (see Online Methods: 'Perturbation of cellular forces with drugs'). The drug was added at t = 0. Droplets rounded up as a consequence on myosin II inhibition, indicating a substantial decrease in the ability of cells to generate forces. (MP4 1457 kb)

Effect of actin polymerization inhibition on droplet deformations

Time-lapse showing the effect of actin polymerization inhibition on droplet deformations. Actin polymerization was inhibited using cytochalasin D (see Online Methods: 'Perturbation of cellular forces with drugs'). The drug was added at t = 0. Droplets rounded up as a consequence on actin polymerization inhibition, indicating a substantial decrease in the ability of cells to generate forces. (MP4 1464 kb)

Effect of cell disruption on droplet deformations

Time-lapse showing the effect of cell disruption on droplet deformations. Cells were disrupted with the detergent sodium dodecyl sulfate (see Online Methods: 'Perturbation of cellular forces with drugs'). The drug was added at t = 0. Cell aggregates disassembled completely in the presence of the drug and droplets rounded up immediately as a consequence, indicating that cell-generated forces were causing the droplet deformations. (MP4 1137 kb)

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Campàs, O., Mammoto, T., Hasso, S. et al. Quantifying cell-generated mechanical forces within living embryonic tissues. Nat Methods 11, 183–189 (2014).

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