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

Mutsuo Nuriya, Shun Fukushima, Atsuya Momotake, Takanori Shinotsuka, Masato Yasui, Tatsuo Arai

Research output: Contribution to journalArticle

23 Citations (Scopus)

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.

Original languageEnglish
Article number11557
JournalNature Communications
Volume7
DOIs
Publication statusPublished - 2016 May 9

Fingerprint

Harmonic generation
Photons
harmonic generations
Coloring Agents
dyes
Imaging techniques
Membrane Potentials
photons
Biological Phenomena
Fluorescent Dyes
Organelles
Action Potentials
membranes
Cell Membrane
Calcium
Neurons
Membranes
organelles
Cell membranes
neurons

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemistry(all)
  • Physics and Astronomy(all)

Cite this

Multimodal two-photon imaging using a second harmonic generation-specific dye. / Nuriya, Mutsuo; Fukushima, Shun; Momotake, Atsuya; Shinotsuka, Takanori; Yasui, Masato; Arai, Tatsuo.

In: Nature Communications, Vol. 7, 11557, 09.05.2016.

Research output: Contribution to journalArticle

Nuriya, Mutsuo ; Fukushima, Shun ; Momotake, Atsuya ; Shinotsuka, Takanori ; Yasui, Masato ; Arai, Tatsuo. / Multimodal two-photon imaging using a second harmonic generation-specific dye. In: Nature Communications. 2016 ; Vol. 7.
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