Bergermann Fabian, Alber Lucas, Sahl Steffen J, Engelhardt Johann, Hell Stefan W
Opt Express. 2015 Jan 12;23(1):211-23. doi: 10.1364/OE.23.000211.
Stimulated Emission Depletion (STED) nanoscopy enables multi-color fluorescence imaging at the nanometer scale. Its typical single-point scanning implementation can lead to long acquisition times. In order to unleash the full spatiotemporal resolution potential of STED nanoscopy, parallelized scanning is mandatory. Here we present a dual-color STED nanoscope utilizing two orthogonally crossed standing light waves as a fluorescence switch-off pattern, and providing a resolving power down to 30 nm. We demonstrate the imaging capabilities in a biological context for immunostained vimentin fibers in a circular field of view of 20 µm diameter at 2000-fold parallelization (i.e. 2000 "intensity minima"). The technical feasibility of massively parallelizing STED without significant compromises in resolution heralds video-rate STED nanoscopy of large fields of view, pending the availability of suitable high-speed detectors.
受激发射损耗（STED）纳米显微镜能够实现纳米级的多色荧光成像。其典型的单点扫描方式可能导致采集时间过长。为了充分发挥STED纳米显微镜的时空分辨率潜力，并行扫描是必不可少的。在此，我们展示了一种双色STED纳米显微镜，它利用两个正交交叉的驻波光作为荧光关闭图案，分辨率可达30纳米。我们在生物学背景下展示了其成像能力，即在直径为20 µm的圆形视野中对免疫染色的波形蛋白纤维进行成像，并行度达到2000倍（即2000个“强度最小值”）。在不显著降低分辨率的情况下大规模并行化STED的技术可行性预示着大视野视频速率STED纳米显微镜的实现，这有待合适的高速探测器的出现。
