Relocating Single Molecules in Super-Resolved Fluorescence Lifetime Images near a Plasmonic Nanostructure
Relocating Single Molecules in Super-Resolved Fluorescence Lifetime Images near a Plasmonic Nanostructure
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DOI：
10.1021/acsphotonics.9b01317
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发表时间：
2020-02-01
期刊：
影响因子：
7
通讯作者：
Krachmalnicoff, Valentina
中科院分区：
文献类型：
--
作者：
Blanquer, Guillaume;van Dam, Bart;Krachmalnicoff, Valentina
单分子定位显微镜是一种强大的技术，具有巨大的潜力，研究轻物质在纳米尺度上的相互作用。纳米结构环境可以改变单个分子的荧光发射，并且诱导的衰减速率改变可以被恢复以映射局部光学态密度（LDOS）。然而，修改的发射器的点扩散函数（PSF）可能会导致其定位错误，设置一个主要的限制，这种方法的可靠性。在本文中，我们解决了这一问题，同时映射的位置和衰变率的单分子和排序事件的衰变率和PSF的大小。借助于数值模拟，我们能够推断偶极子的取向和检索的真实的位置的错位发射。我们已经应用我们的单分子荧光寿命成像显微镜（smFLIM）的方法来研究的LDOS修改的银纳米线超过10 μ m（2）的视野与单分子定位精度类似于15 nm。这是可能的，由于EMCCD相机和单光子雪崩二极管阵列的组合使用，使多路复用和超分辨荧光寿命成像。
Single-molecule localization microscopy is a powerful technique with vast potential to study light matter interactions at the nanoscale. Nanostructured environments can modify the fluorescence emission of single molecules, and the induced decay-rate modification can be retrieved to map the local density of optical states (LDOS). However, the modification of the emitter's point spread function (PSF) can lead to its mislocalization, setting a major limitation to the reliability of this approach. In this paper, we address this by simultaneously mapping the position and decay rate of single molecules and by sorting events by their decay rate and PSF size. With the help of numerical simulations, we are able to infer the dipole orientation and to retrieve the real position of mislocalized emitters. We have applied our approach of single-molecule fluorescence lifetime imaging microscopy (smFLIM) to study the LDOS modification of a silver nanowire over a field of view of 10 mu m(2) with a single-molecule localization precision of similar to 15 nm. This is possible thanks to the combined use of an EMCCD camera and an array of single-photon avalanche diodes, enabling multiplexed and super-resolved fluorescence lifetime imaging.