Advanced easySTED microscopy based on two-photon excitation by electrical modulations of light pulse wavefronts.
Advanced easySTED microscopy based on two-photon excitation by electrical modulations of light pulse wavefronts.
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DOI：
10.1364/boe.9.002671
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发表时间：
2018-06-01
影响因子：
3.4
通讯作者：
Nemoto T
中科院分区：
文献类型：
--
作者：
Otomo K;Hibi T;Fang YC;Hung JH;Tsutsumi M;Kawakami R;Yokoyama H;Nemoto T
我们开发了一种紧凑的受激发射耗尽（STED）双光子激发显微镜，利用电控组件。直接插入物镜透镜前面的透射液晶装置将STED光转换成光学涡旋，同时使激发光不受影响。利用半导体激光器光源产生1.06和0.64 μm两种不同颜色的光脉冲，并灵活控制两个脉冲之间的延迟时间，使荧光抑制比达到最大。在我们的实验中，该系统的空间分辨率比没有STED光照射的情况下高出三倍，并且我们成功地可视化了固定的哺乳动物细胞中的精细微管网络结构，而不会造成显着的光损伤。
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.