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Vertical nanopillars for in situ probing of nuclear mechanics in adherent cells

Nature Nanotechnology volume 10, pages 554–562 (2015)Cite this article

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Abstract

The mechanical stability and deformability of the cell nucleus are crucial to many biological processes, including migration, proliferation and polarization. In vivo, the cell nucleus is frequently subjected to deformation on a variety of length and time scales, but current techniques for studying nuclear mechanics do not provide access to subnuclear deformation in live functioning cells. Here we introduce arrays of vertical nanopillars as a new method for the in situ study of nuclear deformability and the mechanical coupling between the cell membrane and the nucleus in live cells. Our measurements show that nanopillar-induced nuclear deformation is determined by nuclear stiffness, as well as opposing effects from actin and intermediate filaments. Furthermore, the depth, width and curvature of nuclear deformation can be controlled by varying the geometry of the nanopillar array. Overall, vertical nanopillar arrays constitute a novel approach for non-invasive, subcellular perturbation of nuclear mechanics and mechanotransduction in live cells.

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Figure 1: The nuclear envelope deforms around nanopillars, creating a non-invasive platform for studying in situ nuclear deformation.
Figure 2: Quantitative analysis of nanopillar-induced nuclear deformation.
Figure 3: Nanopillar-induced nuclear deformation shows a strong dependence on nuclear stiffness.
Figure 4: Nanopillar-induced nuclear deformation depends on the integrity of cytoskeletal components.
Figure 5: Nanopillar geometry determines the depth and shape of nuclear deformation.
Figure 6: Finite-element analysis suggests that nanopillar-induced deformation is a cumulative result of both the actin cap above the nucleus and actin accumulated around the nanopillar.

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Acknowledgements

This work was supported by the National Science Foundation (award no. 1055112 and 1344302), the National Institutes of Health (grant no. DP2NS082125), a Searle Scholar award, a Packard Science and Engineering Fellowship (to B.C.) and a National Defense Science and Engineering Graduate Fellowship (to Z.L.). The authors thank H. Worman at Columbia University for providing the GFP-Sun2 plasmid. The authors thank M. Lin at Stanford University for helping with the confocol microscope.

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Authors and Affiliations

  1. Department of Chemistry, 333 Campus Drive, Stanford, 94305, California, USA

    Lindsey Hanson, Hsin-Ya Lou, Praveen Chowdary & Bianxiao Cui

  2. Department of Materials Science and Engineering, 496 Lomita Mall, Stanford, 94305, California, USA

    Wenting Zhao, Seok Woo Lee & Yi Cui

  3. Department of Applied Physics, 348 Via Pueblo, Stanford University, Stanford, 94305, California, USA

    Ziliang Carter Lin

  4. Stanford Institute for Materials and Energy Sciences, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025, California, USA

    Yi Cui

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Contributions

L.H., B.C. and Y.C. designed experiments. L.H., W.Z., H.L. and Z.L. carried out the experiments. L.H. and S.W.L. designed and carried out the simulations. L.H. and P.C. designed and carried out the data analysis. All authors contributed to scientific planning, discussions and writing of the manuscript.

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Correspondence to Yi Cui or Bianxiao Cui.

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Hanson, L., Zhao, W., Lou, HY. et al. Vertical nanopillars for in situ probing of nuclear mechanics in adherent cells. Nature Nanotech 10, 554–562 (2015). https://doi.org/10.1038/nnano.2015.88

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