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Multimodal two-photon imaging using a second harmonic generation-specific dye

Author

Listed:
  • Mutsuo Nuriya

    (Keio University
    Graduate School of Environment and Information Sciences, Yokohama National University)

  • Shun Fukushima

    (Graduate School of Pure and Applied Sciences and Tsukuba Research Center for Interdisciplinary Materials Science (TIMS), University of Tsukuba)

  • Atsuya Momotake

    (Graduate School of Pure and Applied Sciences and Tsukuba Research Center for Interdisciplinary Materials Science (TIMS), University of Tsukuba)

  • Takanori Shinotsuka

    (Keio University)

  • Masato Yasui

    (Keio University)

  • Tatsuo Arai

    (Graduate School of Pure and Applied Sciences and Tsukuba Research Center for Interdisciplinary Materials Science (TIMS), University of Tsukuba)

Abstract

Second harmonic generation (SHG) imaging can be used to visualize unique biological phenomena, but currently available dyes limit its application owing to the strong fluorescent signals that they generate together with SHG. Here we report the first non-fluorescent and membrane potential-sensitive SHG-active organic dye Ap3. Ap3 is photostable and generates SH signals at the plasma membrane with virtually no fluorescent signals, in sharp contrast to the previously used fluorescent dye FM4-64. When tested in neurons, Ap3-SHG shows linear membrane potential sensitivity and fast responses to action potentials, and also shows significantly reduced photodamage compared with FM4-64. The SHG-specific nature of Ap3 allows simultaneous and completely independent imaging of SHG signals and fluorescent signals from various reporter molecules, including markers of cellular organelles and intracellular calcium. Therefore, this SHG-specific dye enables true multimodal two-photon imaging in biological samples.

Suggested Citation

  • Mutsuo Nuriya & Shun Fukushima & Atsuya Momotake & Takanori Shinotsuka & Masato Yasui & Tatsuo Arai, 2016. "Multimodal two-photon imaging using a second harmonic generation-specific dye," Nature Communications, Nature, vol. 7(1), pages 1-10, September.
    • Handle: RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms11557
    DOI: 10.1038/ncomms11557
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