Elastomeric sensor surfaces for high-throughput single-cell force cytometry

Nat Biomed Eng. 2018 Feb;2(2):124-137. doi: 10.1038/s41551-018-0193-2. Epub 2018 Feb 6.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Asthma / pathology
  • Cell Differentiation
  • Cells, Cultured
  • Elastomers / chemistry*
  • Fluorescent Dyes / chemistry
  • Formoterol Fumarate / pharmacology
  • Heterocyclic Compounds, 4 or More Rings / pharmacology
  • Humans
  • Macrophages / cytology
  • Macrophages / metabolism
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / metabolism
  • Microscopy, Fluorescence
  • Myocardial Contraction / drug effects
  • Myocytes, Smooth Muscle / cytology
  • Myocytes, Smooth Muscle / metabolism
  • Phagocytosis / drug effects
  • Single-Cell Analysis / methods*

Substances

  • Elastomers
  • Fluorescent Dyes
  • Heterocyclic Compounds, 4 or More Rings
  • blebbistatin
  • Formoterol Fumarate