Moneron Gael, Medda Rebecca, Hein Birka, Giske Arnold, Westphal Volker, Hell Stefan W
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
Opt Express. 2010 Jan 18;18(2):1302-9. doi: 10.1364/OE.18.001302.
We report on fast beam-scanning stimulated-emission-depletion (STED) microscopy in the visible range using for resolution enhancement compact, low cost and turn-key continuous wave (CW) fiber lasers emitting at 592 nm. Spatial resolutions of 35 to 65 nm in the focal plane are shown for various samples including fluorescent nanoparticles, immuno-stained cells with a non-exhaustive selection of 5 commonly used organic fluorescent markers, and living cells expressing the yellow fluorescent protein Citrine. The potential of the straightforward combination of CW-STED and fast beam scanning is illustrated in a movie of the endoplasmic reticulum (ER) of a living cell, composed of 100 frames (6 microm x 12 microm), each of them acquired in a time shorter than 0.2 s.
我们报道了在可见光范围内的快速光束扫描受激发射损耗（STED）显微镜技术，该技术使用紧凑、低成本且交钥匙式的连续波（CW）光纤激光器来增强分辨率，这些激光器在592 nm波长处发射。对于各种样品，包括荧光纳米颗粒、用5种常用有机荧光标记物进行非详尽选择免疫染色的细胞以及表达黄色荧光蛋白柠檬黄的活细胞，展示了焦平面上35至65 nm的空间分辨率。连续波STED与快速光束扫描的直接结合的潜力在一个活细胞内质网（ER）的视频中得到了体现，该视频由100帧（6微米×12微米）组成，每帧的采集时间短于0.2秒。
