School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, China.
PLoS One. 2012;7(6):e40003. doi: 10.1371/journal.pone.0040003. Epub 2012 Jun 27.
In this report, a Ti:Sapphire oscillator was utilized to realize synchronization-free stimulated emission depletion (STED) microscopy. With pump power of 4.6 W and sample irradiance of 310 mW, we achieved super-resolution as high as 71 nm. With synchronization-free STED, we imaged 200 nm nanospheres as well as all three cytoskeletal elements (microtubules, intermediate filaments, and actin filaments), clearly demonstrating the resolving power of synchronization-free STED over conventional diffraction limited imaging. It also allowed us to discover that, Dylight 650, exhibits improved performance over ATTO647N, a fluorophore frequently used in STED. Furthermore, we applied synchronization-free STED to image fluorescently-labeled intracellular viral RNA granules, which otherwise cannot be differentiated by confocal microscopy. Thanks to the widely available Ti:Sapphire oscillators in multiphoton imaging system, this work suggests easier access to setup super-resolution microscope via the synchronization-free STED.
在本报告中，我们利用掺钛蓝宝石振荡器实现了无需同步的受激发射损耗（STED）显微镜。在 4.6 W 的泵浦功率和 310 mW 的样品辐照度下，我们实现了高达 71nm 的超分辨率。通过无需同步的 STED，我们成功地对 200nm 纳米球以及三种细胞骨架元件（微管、中间丝和肌动蛋白丝）进行了成像，清晰地证明了无需同步的 STED 相对于传统的衍射极限成像的分辨率优势。此外，我们还发现，Dylight 650 比常用于 STED 的荧光染料 ATTO647N 具有更好的性能。此外，我们还应用无需同步的 STED 对荧光标记的细胞内病毒 RNA 颗粒进行了成像，这些颗粒在共聚焦显微镜下无法区分。由于多光子成像系统中广泛使用掺钛蓝宝石振荡器，本工作表明通过无需同步的 STED 更容易实现超分辨率显微镜的搭建。
