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Acoustic tweezers for high-throughput single-cell analysis

Nature Protocols volume 18, pages 2441–2458 (2023)Cite this article

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Abstract

Acoustic tweezers provide an effective means for manipulating single cells and particles in a high-throughput, precise, selective and contact-free manner. The adoption of acoustic tweezers in next-generation cellular assays may advance our understanding of biological systems. Here we present a comprehensive set of instructions that guide users through device fabrication, instrumentation setup and data acquisition to study single cells with an experimental throughput that surpasses traditional methods, such as atomic force microscopy and micropipette aspiration, by several orders of magnitude. With acoustic tweezers, users can conduct versatile experiments that require the trapping, patterning, pairing and separation of single cells in a myriad of applications ranging across the biological and biomedical sciences. This procedure is widely generalizable and adaptable for investigations in materials and physical sciences, such as the spinning motion of colloids or the development of acoustic-based quantum simulations. Overall, the device fabrication requires ~12 h, the experimental setup of the acoustic tweezers requires 1–2 h and the cell manipulation experiment requires ~30 min to complete. Our protocol is suitable for use by interdisciplinary researchers in biology, medicine, engineering and physics.

Key points

  • The procedural steps cover the fabrication of the microfluidic chamber, the interdigital transducer and the acoustic tweezers, followed by the experimental setup, the culturing of cells and their chemically induced perturbations and the manipulation of single cells and particles, data acquisition and analysis.

  • Acoustic tweezers enable the fast, high-throughput handling of single cells and particles, and outperforms alternative methods such as atomic force microscopy, optical tweezers or micropipettes.

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Fig. 1: Flow diagram of the workflow.
Fig. 2: Device fabrication and experimental setup of the acoustic tweezers.
Fig. 3: Cell culture and chemical perturbations.
Fig. 4: Anticipated results with the acoustic tweezers.

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Data availability

The main data discussed in this protocol are available in the supporting primary research paper30. The raw datasets are available for research purposes from the corresponding authors upon reasonable request.

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Acknowledgements

We acknowledge support from the Shared Materials Instrumentation Facility at Duke University. We acknowledge support from the National Institutes of Health (R01GM145960 (L.P.L), R01GM141055 (T.J.H.), R01GM132603 (T.J.H.), R01HD103727 (T.J.H.), R01GM143439 (T.J.H.), R01GM135486 (T.J.H.), R44AG063643 (T.J.H.), R44OD024963 (T.J.H.), R44HL140800 (T.J.H.), R21HD102790 (T.J.H.), U18TR003778 (T.J.H.) and UH3TR002978 (T.J.H.)), the National Science Foundation (ECCS-1807601 (T.J.H.), MCB-2042704 (T.J.H.) and CMMI-2104295 (T.J.H.)) and the National Science Foundation Graduate Research Fellowship (1644868 (J.R.)).

Author information

Authors and Affiliations

  1. Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC, USA

    Shujie Yang, Joseph Rufo, Ruoyu Zhong, Zeyu Wang & Tony Jun Huang

  2. Department of Biomedical Engineering, Duke University, Durham, NC, USA

    Joseph Rich

  3. Renal Division and Division of Engineering in Medicine, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Luke P. Lee

  4. Department of Bioengineering, Department of Electrical Engineering and Computer Science, University of California, Berkeley, Berkeley, CA, USA

    Luke P. Lee

  5. Institute of Quantum Biophysics, Department of Biophysics, Sungkyunkwan University, Suwon, South Korea

    Luke P. Lee

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Contributions

S.Y., L.P.L. and T.J.H. designed the research. S.Y. performed the research. Z.W. performed the western blot analysis. S.Y., J. Rufo, R.Z., J. Rich, L.P.L., Z. W. and T.J.H. analyzed data. S.Y., R.Z. and L.P.L. drew the figures. S.Y., J. Rufo, J. Rich, L.P.L. and T.J.H. wrote the paper. S.Y., J. Rich, L.P.L. and T.J.H. revised the manuscript.

Corresponding authors

Correspondence to Luke P. Lee or Tony Jun Huang.

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Competing interests

T.J.H. has cofounded a start-up company, Ascent Bio-Nano Technologies Inc., to commercialize technologies involving acoustofluidics and acoustic tweezers. The remaining authors declare no competing interests.

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Nature Protocols thanks Weiwei Cui, Kishan Dholakia, Tiziano Serra and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Related links

Yang, S. et al. Nat. Mater. 21, 540–546 (2022): https://doi.org/10.1038/s41563-022-01210-8

Supplementary information

Supplementary Information

Supplementary Figs. 1–4, Table 1, Notes 1 and 2 and Appendix 1.

Reporting Summary

Supplementary Video 1

Fabrication process of the acoustic tweezers.

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Yang, S., Rufo, J., Zhong, R. et al. Acoustic tweezers for high-throughput single-cell analysis. Nat Protoc 18, 2441–2458 (2023). https://doi.org/10.1038/s41596-023-00844-5

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