Vicidomini Giuseppe, Hernández Ivan Coto, d'Amora Marta, Zanacchi Francesca Cella, Bianchini Paolo, Diaspro Alberto
Department of Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genoa, Italy.
Department of Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genoa, Italy; Department of Physics, University of Genoa, Via Dodecaneso 33, 16146 Genoa, Italy.
Methods. 2014 Mar 15;66(2):124-30. doi: 10.1016/j.ymeth.2013.06.029. Epub 2013 Jun 29.
Stimulation emission depletion (STED) microscopy breaks the spatial resolution limit of conventional light microscopy while retaining its major advantages, such as working under physiological conditions. These properties make STED microscopy a perfect tool for investigating dynamic sub-cellular processes in living organisms. However, up to now, the massive dissemination of STED microscopy has been hindered by the complexity and cost of its implementation. Gated CW-STED (gCW-STED) substantially helps solve this problem without sacrificing spatial resolution. Here, we describe a versatile gCW-STED microscope able to speedily image the specimen, at a resolution below 50 nm, with light intensities comparable to the more complicated all-pulsed STED system. We show this ability on calibration samples as well as on biological samples.
受激发射损耗（STED）显微镜突破了传统光学显微镜的空间分辨率限制，同时保留了其主要优点，如可在生理条件下工作。这些特性使STED显微镜成为研究活生物体中动态亚细胞过程的理想工具。然而，到目前为止，STED显微镜的广泛应用一直受到其实施复杂性和成本的阻碍。门控连续波STED（gCW-STED）在不牺牲空间分辨率的情况下极大地有助于解决这一问题。在此，我们描述了一种通用的gCW-STED显微镜，它能够以低于50纳米的分辨率对样品进行快速成像，其光强度与更复杂的全脉冲STED系统相当。我们在校准样品以及生物样品上展示了这种能力。
