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

作者:Otomo Kohei; Hibi Terumasa; Fang Yi Cheng; Hung Jui Hung; Tsutsumi Motosuke; Kawakami Ryosuke; Yokoyama Hiroyuki; Nemoto Tomomi*
来源:Biomedical Optics Express, 2018, 9(6): 2671-2680.
DOI:10.1364/BOE.9.002671

摘要

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 mu m, 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.

  • 出版日期2018-6-1