As cells with aberrant force-generating phenotypes can directly lead to disease, cellular force-generation mechanisms are high-value targets for new therapies. Here, we show that single-cell force sensors embedded in elastomers enable single-cell force measurements with ~100-fold improvement in throughput than was previously possible. The microtechnology is scalable and seamlessly integrates with the multi-well plate format, enabling highly parallelized time-course studies. In this regard, we show that airway smooth muscle cells isolated from fatally asthmatic patients have innately greater and faster force-generation capacity in response to stimulation than healthy control cells. By simultaneously tracing agonist-induced calcium flux and contractility in the same cell, we show that the calcium level is ultimately a poor quantitative predictor of cellular force generation. Finally, by quantifying phagocytic forces in thousands of individual human macrophages, we show that force initiation is a digital response (rather than a proportional one) to the proper immunogen. By combining mechanobiology at the single-cell level with high-throughput capabilities, this microtechnology can support drug-discovery efforts for clinical conditions associated with aberrant cellular force generation.

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  • Correction 12 March 2018

    In the version of this Article originally published, in Fig. 1a, all cells in the top schematic were missing, and in the bottom-left schematic showing multiple pattern shapes, two cells were missing in the bottom-right corner. This figure has now been updated in all versions of the Article.


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The work was supported by the National Institutes of Health Director’s New Innovator Award 1DP2OD007113, the David and Lucile Packard Fellowship and the National Institutes of Health/National Institute of Biomedical Imaging and Bioengineering R21 Award 1R21EB024081-01. The authors thank C. Walthers for providing the primary mouse intestinal SMCs used in the Supplementary Videos, Y. Wang and J. Z. Lee for providing the neonatal rat ventricular myocytes, and O. Adeyiga for performing bloods draws over the course of six months to facilitate primary macrophage culture. All microfabrication steps were completed using equipment provided by the Integrated Systems Nanofabrication Cleanroom at the California NanoSystems Institute at the University of California, Los Angeles.

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Author notes

  1. These authors contributed equally: Ivan Pushkarsky and Peter Tseng.


  1. Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA, USA

    • Ivan Pushkarsky
    • , Peter Tseng
    • , Dylan Black
    • , Lyndon Warfe
    • , Jonathan Lin
    •  & Dino Di Carlo
  2. Department of Electrical Engineering and Computer Science, University of California, Los Angeles, Irvine, CA, USA

    • Peter Tseng
  3. California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA, USA

    • Bryan France
    • , Robert Damoiseaux
    •  & Dino Di Carlo
  4. Rutgers Institute for Translational Medicine and Science, Child Health Institute, Rutgers University, New Brunswick, NJ, USA

    • Cynthia J. Koziol-White
    • , William F. Jester Jr
    •  & Reynold A. Panettieri Jr
  5. Department of Microbiology, Immunology and Molecular Genetics and The Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, USA

    • Ryan K. Trinh
    •  & Sherie L. Morrison
  6. Division of Dermatology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA

    • Philip O. Scumpia
  7. Department of Molecular and Medicinal Pharmacology, University of California, Los Angeles, Los Angeles, CA, USA

    • Robert Damoiseaux
  8. Department of Mechanical Engineering, University of California, Los Angeles, Los Angeles, CA, USA

    • Dino Di Carlo


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P.T., D.D.C. and I.P. conceived the methods. I.P., R.D., P.O.S., S.L.M., R.A.P., C.J.K.-W. and D.D.C. designed the experiments. I.P. performed all the experiments, developed the multi-well embodiment, optimized the protocols and wrote the image analysis software. D.B. assisted with the substrate preparation and macrophage differentiation procedures. L.W. assisted with the substrate preparation. R.K.T. maintained the chimeric antibody stocks. S.L.M. supplied all the chimeric antibodies. J.L. constructed the finite element method model. R.D. supplied the high-throughput screening (HTS) equipment for dose-response experiments and provided extensive guidance and technical advice on HTS procedures and developing the multi-well plate embodiment. B.F. assisted with the HTS equipment and drug administration. W.F.J. maintained the donor HASM cells. P.O.S. performed the macrophage and dendritic cells differentiation and advised the experimental procedures. I.P., R.D., P.O.S., S.L.M., R.A.P., C.J.K.-W. and D.D.C. interpreted the results. I.P. and D.D.C. wrote the manuscript. R.D., P.O.S., S.L.M., R.A.P. and C.J.K.-W. helped revise the manuscript.

Competing interests

I.P., P.T. and D.D.C. are named inventors on a patent application by the University of California, Los Angeles that covers the technology described in this study. I.P., R.D. and D.D.C. have a financial interest in Forcyte Biotechnologies, which aims to commercialize FLECS technology.

Corresponding author

Correspondence to Dino Di Carlo.

Supplementary information

  1. Supplementary Information

    Supplementary figures, tables and video captions.

  2. Life Sciences Reporting Summary

  3. Supplementary Text

    MATLAB computer code used to evaluate micropattern displacements and calcium dye intensity.


  1. Supplementary Video 1

    Fibronectin–fibrinogen patterns created using our sacrificial dextran method.

  2. Supplementary Video 2

    A second example of fibronectin–fibrinogen patterns created using the sacrificial dextran method.

  3. Supplementary Video 3

    Patterns contracted by adhered HeLa cells relax after the addition of the myosin inhibitor blebbistatin. Time-lapsed video.

  4. Supplementary Video 4

    HASM cells treated with 1 μM bradykinin contract significantly beyond tonic levels within 30 minutes.

  5. Supplementary Video 5

    HASM cells treated with 100 nM endothelin-1 contract significantly beyond tonic levels within 30 minutes.

  6. Supplementary Video 6

    Visualization of calcium flux within HASM cells seeded on arrays of FLECS micropatterns after the addition of Hist.

  7. Supplementary Video 7

    Time-lapsed video of human monocyte-derived macrophages engaged in frustrated phagocytosis of an IgG-opsonized patterned surface.

  8. Supplementary Video 8

    Representative video (real time) of a phasically contracting neonatal rat ventricular myocyte.

  9. Supplementary Video 9

    Representative video (real time) of a pattern phasically contracted by an adhered and spontaneously beating neonatal rat ventricular myocyte.

  10. Supplementary Video 10

    Representative video (real time) of a pattern phasically contracted by an adhered and beating neonatal rat ventricular myocyte paced with pulsed electric fields at frequencies of 1 Hz and 2 Hz.

  11. Supplementary Video 11

    Fibronectin–fibrinogen patterns created using adsorption rather than our sacrificial dextran method. Time-lapse video.

  12. Supplementary Video 12

    Second example of fibronectin–fibrinogen patterns created using adsorption rather than our sacrificial dextran method. Time-lapse video.

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