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Automated single-molecule imaging in living cells

Author

Listed:
  • Masato Yasui

    (Laboratory for Cell Signaling Dynamics, RIKEN BDR)

  • Michio Hiroshima

    (Laboratory for Cell Signaling Dynamics, RIKEN BDR
    Cellular Informatics Laboratory, RIKEN)

  • Jun Kozuka

    (Laboratory for Cell Signaling Dynamics, RIKEN BDR)

  • Yasushi Sako

    (Cellular Informatics Laboratory, RIKEN)

  • Masahiro Ueda

    (Laboratory for Cell Signaling Dynamics, RIKEN BDR
    Osaka University)

Abstract

An automated single-molecule imaging system developed for live-cell analyses based on artificial intelligence-assisted microscopy is presented. All significant procedures, i.e., searching for cells suitable for observation, detecting in-focus positions, and performing image acquisition and single-molecule tracking, are fully automated, and numerous highly accurate, efficient, and reproducible single-molecule imaging experiments in living cells can be performed. Here, the apparatus is applied for single-molecule imaging and analysis of epidermal growth factor receptors (EGFRs) in 1600 cells in a 96-well plate within 1 day. Changes in the lateral mobility of EGFRs on the plasma membrane in response to various ligands and drug concentrations are clearly detected in individual cells, and several dynamic and pharmacological parameters are determined, including the diffusion coefficient, oligomer size, and half-maximal effective concentration (EC50). Automated single-molecule imaging for systematic cell signaling analyses is feasible and can be applied to single-molecule screening, thus extensively contributing to biological and pharmacological research.

Suggested Citation

  • Masato Yasui & Michio Hiroshima & Jun Kozuka & Yasushi Sako & Masahiro Ueda, 2018. "Automated single-molecule imaging in living cells," Nature Communications, Nature, vol. 9(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-05524-7
    DOI: 10.1038/s41467-018-05524-7
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    Cited by:

    1. Daisuke Watanabe & Michio Hiroshima & Masato Yasui & Masahiro Ueda, 2024. "Single molecule tracking based drug screening," Nature Communications, Nature, vol. 15(1), pages 1-14, December.
    2. Michael G. Sugiyama & Aidan I. Brown & Jesus Vega-Lugo & Jazlyn P. Borges & Andrew M. Scott & Khuloud Jaqaman & Gregory D. Fairn & Costin N. Antonescu, 2023. "Confinement of unliganded EGFR by tetraspanin nanodomains gates EGFR ligand binding and signaling," Nature Communications, Nature, vol. 14(1), pages 1-17, December.
    3. Tatsat Banerjee & Satomi Matsuoka & Debojyoti Biswas & Yuchuan Miao & Dhiman Sankar Pal & Yoichiro Kamimura & Masahiro Ueda & Peter N. Devreotes & Pablo A. Iglesias, 2023. "A dynamic partitioning mechanism polarizes membrane protein distribution," Nature Communications, Nature, vol. 14(1), pages 1-24, December.

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