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

Evan P. Perillo, Yen-Liang Liu, Khang Huynh, Cong Liu, Chao-Kai Chou, Mien-Chie Hung, Hsin-Chih Yeh () and Andrew K. Dunn ()
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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
Mien-Chie Hung: The University of Texas MD Anderson Cancer Center
Hsin-Chih Yeh: The University of Texas at Austin
Andrew K. Dunn: The University of Texas at Austin

Nature Communications, 2015, vol. 6, issue 1, 1-8

Abstract: 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.

Date: 2015
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DOI: 10.1038/ncomms8874

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