Two-photon STED nanoscopy realizing 100-nm spatial resolution utilizing high-peak-power sub-nanosecond 655-nm pulses
Two-photon STED nanoscopy realizing 100-nm spatial resolution utilizing high-peak-power sub-nanosecond 655-nm pulses
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DOI：
10.1364/boe.10.003104
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发表时间：
2019-07-01
影响因子：
3.4
通讯作者：
Nemoto, Tomomi
中科院分区：
文献类型：
--
作者：
Ishii, Hirokazu;Otomo, Kohei;Nemoto, Tomomi
我们利用高峰值功率的亚纳秒655 nm脉冲，开发了双光子激发受激发射耗尽（STED）纳米显微镜。STED脉冲具有理想的光学特性和足够的脉冲能量，在具有可控元件的紧凑设置中实现了70 nm的空间分辨率。在生物学应用方面，我们筛选了合适的荧光染料或蛋白质，实现了固定原代培养神经元突触前蛋白簇的亚100 nm空间分辨率成像，而没有严重的光漂白。我们期望这种方法能够使代表活细胞和组织生理功能的生物分子的超微结构和簇动力学可视化。(C) 2019年美国光学学会根据OSA开放获取出版协议的条款
We developed two-photon excitation stimulated emission depletion (STED) nanoscopy using high-peak-power sub-nanosecond 655-nm pulses. The STED pulse exhibited ideal optical properties and sufficient pulse energy to realize a 70-nm spatial resolution in the compact setup With electrically controllable components. For biological applications, we screened suitable fluorescent dyes or proteins and realized the sub-100 nm spatial resolution imaging of presynaptic protein clusters in fixed primary cultured neurons Without severe photobleaching. We expect this method to enable visualization of ultrastructures and the cluster dynamics of biomolecules representing physiological functions in living cells and tissue. (C) 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement