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Deep and high-resolution three-dimensional tracking of single particles using nonlinear and multiplexed illumination

Author

Listed:
  • Evan P. Perillo

    (The University of Texas at Austin)

  • Yen-Liang Liu

    (The University of Texas at Austin)

  • Khang Huynh

    (The University of Texas at Austin)

  • Cong Liu

    (The University of Texas at Austin)

  • Chao-Kai Chou

    (The University of Texas MD Anderson Cancer Center
    Center for Molecular Medicine and Graduate Institute of Cancer Biology, China Medical University)

  • Mien-Chie Hung

    (The University of Texas MD Anderson Cancer Center
    Center for Molecular Medicine and Graduate Institute of Cancer Biology, China Medical University)

  • Hsin-Chih Yeh

    (The University of Texas at Austin)

  • Andrew K. Dunn

    (The University of Texas at Austin)

Abstract

Molecular trafficking within cells, tissues and engineered three-dimensional multicellular models is critical to the understanding of the development and treatment of various diseases including cancer. However, current tracking methods are either confined to two dimensions or limited to an interrogation depth of ∼15 μm. Here we present a three-dimensional tracking method capable of quantifying rapid molecular transport dynamics in highly scattering environments at depths up to 200 μm. The system has a response time of 1 ms with a temporal resolution down to 50 μs in high signal-to-noise conditions, and a spatial localization precision as good as 35 nm. Built on spatiotemporally multiplexed two-photon excitation, this approach requires only one detector for three-dimensional particle tracking and allows for two-photon, multicolour imaging. Here we demonstrate three-dimensional tracking of epidermal growth factor receptor complexes at a depth of ∼100 μm in tumour spheroids.

Suggested Citation

  • Evan P. Perillo & Yen-Liang Liu & Khang Huynh & Cong Liu & Chao-Kai Chou & Mien-Chie Hung & Hsin-Chih Yeh & Andrew K. Dunn, 2015. "Deep and high-resolution three-dimensional tracking of single particles using nonlinear and multiplexed illumination," Nature Communications, Nature, vol. 6(1), pages 1-8, November.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms8874
    DOI: 10.1038/ncomms8874
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    Cited by:

    1. Larissa Kohler & Matthias Mader & Christian Kern & Martin Wegener & David Hunger, 2021. "Tracking Brownian motion in three dimensions and characterization of individual nanoparticles using a fiber-based high-finesse microcavity," Nature Communications, Nature, vol. 12(1), pages 1-7, December.
    2. Elias Amselem & Bo Broadwater & Tora Hävermark & Magnus Johansson & Johan Elf, 2023. "Real-time single-molecule 3D tracking in E. coli based on cross-entropy minimization," Nature Communications, Nature, vol. 14(1), pages 1-11, December.
    3. K. S. Vikrant & G. R. Jayanth, 2022. "Diamagnetically levitated nanopositioners with large-range and multiple degrees of freedom," Nature Communications, Nature, vol. 13(1), pages 1-11, December.
    4. Andrea Bucci & Giorgio Tortarolo & Marcus Oliver Held & Luca Bega & Eleonora Perego & Francesco Castagnetti & Irene Bozzoni & Eli Slenders & Giuseppe Vicidomini, 2024. "4D Single-particle tracking with asynchronous read-out single-photon avalanche diode array detector," Nature Communications, Nature, vol. 15(1), pages 1-12, December.

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