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High-throughput ballistic injection nanorheology to measure cell mechanics

Nature Protocols volume 7, pages 155–170 (2012)Cite this article

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Abstract

High-throughput ballistic injection nanorheology is a method for the quantitative study of cell mechanics. Cell mechanics are measured by ballistic injection of submicron particles into the cytoplasm of living cells and tracking the spontaneous displacement of the particles at high spatial resolution. The trajectories of the cytoplasm-embedded particles are transformed into mean-squared displacements, which are subsequently transformed into frequency-dependent viscoelastic moduli and time-dependent creep compliance of the cytoplasm. This method allows for the study of a wide range of cellular conditions, including cells inside a 3D matrix, cell subjected to shear flows and biochemical stimuli, and cells in a live animal. Ballistic injection lasts <1 min and is followed by overnight incubation. Multiple particle tracking for one cell lasts <1 min. Forty cells can be examined in <1 h.

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Figure 1: Minimal cell death is induced by ballistic injection of nanoparticles into the cytoplasm of adherent cells.
Figure 2: Biolistic machine used to introduce submicron particles inside cells for high-throughout ballistic nanorheology (htBIN) analysis.
Figure 3: Image processing steps taken to enhance the high-resolution tracking of fluorescent nanoparticles in the cytoplasm of live cells.
Figure 4: htBIN analysis of normal and cancer human ovarian epithelial cells.

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Acknowledgements

Normal human ovarian epithelial cells (OSE10) and ovarian cancer cells (OVCAR3) were provided by I.-M. Shih (Department of Pathology, Johns Hopkins University School of Medicine). This work was supported in part by NIH grants U54CA143868 and R21CA137686, and RO1GM084204. We thank members of the Wirtz and Tseng labs for technical advice and reagents.

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Author notes

  1. Pei-Hsun Wu and Christopher M Hale: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, The Johns Hopkins University, Baltimore, Maryland, USA

    Pei-Hsun Wu, Christopher M Hale, Wei-Chiang Chen, Jerry S H Lee & Denis Wirtz

  2. Johns Hopkins Physical Sciences-Oncology Center, The Johns Hopkins University, Baltimore, Maryland, USA

    Pei-Hsun Wu, Christopher M Hale, Wei-Chiang Chen, Yiider Tseng & Denis Wirtz

  3. Center for Strategic Scientific Initiatives, Office of the Director, National Cancer Institute, US National Institutes of Health (NIH), Bethesda, Maryland, USA

    Jerry S H Lee

  4. Department of Chemical Engineering, University of Florida, Gainesville, Florida, USA

    Yiider Tseng

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Contributions

P.-H.W., C.M.H. and W.-C.C. conducted experiments. P.-H.W., C.M.H., J.S.H.L., Y.T. and D.W. designed the experiments, analyzed the results and wrote the paper.

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Correspondence to Yiider Tseng or Denis Wirtz.

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Wu, PH., Hale, C., Chen, WC. et al. High-throughput ballistic injection nanorheology to measure cell mechanics. Nat Protoc 7, 155–170 (2012). https://doi.org/10.1038/nprot.2011.436

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