Sidenstein Sven C, D'Este Elisa, Böhm Marvin J, Danzl Johann G, Belov Vladimir N, Hell Stefan W
Max Planck Institute for Biophysical Chemistry, Department of NanoBiophotonics, Am Fassberg 11, 37077 Göttingen, Germany.
Sci Rep. 2016 May 25;6:26725. doi: 10.1038/srep26725.
Superresolution fluorescence microscopy of multiple fluorophores still requires development. Here we present simultaneous three-colour stimulated emission depletion (STED) nanoscopy relying on a single STED beam at 620 nm. Toggling the STED beam between two or more power levels ("multilevelSTED") optimizes resolution and contrast in all colour channels, which are intrinsically co-aligned and well separated. Three-colour recording is demonstrated by imaging the nanoscale cytoskeletal organization in cultured hippocampal neurons. The down to ~35 nm resolution identified periodic actin/betaII spectrin lattices along dendrites and spines; however, at presynaptic and postsynaptic sites, these patterns were found to be absent. Both our multicolour scheme and the 620 nm STED line should be attractive for routine STED microscopy applications.
多荧光团的超分辨率荧光显微镜技术仍有待发展。在此，我们展示了一种基于620nm单一受激发射损耗（STED）光束的同步三色受激发射损耗纳米显微镜技术。在两个或更多功率水平之间切换STED光束（“多级STED”）可优化所有颜色通道中的分辨率和对比度，这些通道在本质上是共对准且分隔良好的。通过对培养的海马神经元中的纳米级细胞骨架组织进行成像，展示了三色记录。低至约35nm的分辨率确定了沿树突和棘的周期性肌动蛋白/βII血影蛋白晶格；然而，在突触前和突触后位点，未发现这些模式。我们的多色方案和620nm STED谱线对于常规STED显微镜应用都应具有吸引力。
