Advanced easySTED microscopy based on two-photon excitation by electrical modulations of light pulse wavefronts

Kohei Otomo, Terumasa Hibi, Yi Cheng Fang, Jui Hung Hung, Motosuke Tsutsumi, Ryosuke Kawakami, Hiroyuki Yokoyama, Tomomi Nemoto

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

We developed a compact stimulated emission depletion (STED) two-photon excitation microscopy that utilized electrically controllable components. Transmissive liquid crystal devices inserted directly in front of the objective lens converted the STED light into an optical vortex while leaving the excitation light unaffected. Light pulses of two different colors, 1.06 and 0.64 μm, were generated by laser diode-based light sources, and the delay between the two pulses was flexibly controlled so as to maximize the fluorescence suppression ratio. In our experiments, the spatial resolution of this system was up to three times higher than that obtained without STED light irradiation, and we successfully visualize the fine microtubule network structures in fixed mammalian cells without causing significant photo-damage.

Original languageEnglish
Article number#318596
Pages (from-to)2671-2680
Number of pages10
JournalBiomedical Optics Express
Volume9
Issue number6
DOIs
Publication statusPublished - 2018 Jun 1

ASJC Scopus subject areas

  • Biotechnology
  • Atomic and Molecular Physics, and Optics

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    Otomo, K., Hibi, T., Fang, Y. C., Hung, J. H., Tsutsumi, M., Kawakami, R., Yokoyama, H., & Nemoto, T. (2018). Advanced easySTED microscopy based on two-photon excitation by electrical modulations of light pulse wavefronts. Biomedical Optics Express, 9(6), 2671-2680. [#318596]. https://doi.org/10.1364/BOE.9.002671