Relocating Single Molecules in Super-Resolved Fluorescence Lifetime Images near a Plasmonic Nanostructure

被引:19
作者
Blanquer, Guillaume [1 ]
van Dam, Bart [1 ]
Gulinatti, Angelo [2 ]
Acconcia, Giulia [2 ]
De Wilde, Yannick [1 ]
Izeddin, Ignacio [1 ]
Krachmalnicoff, Valentina [1 ]
机构
[1] PSL Univ, CNRS, Inst Langevin, ESPCI Paris, F-75005 Paris, France
[2] Politecn Milan, Dipartimento Elettron Informaz & Bioingn, I-20133 Milan, Italy
关键词
single-molecule localization microscopy (SMLM); single-molecule fluorescence lifetime imaging microscopy (smFLIM); point spread function (PSF); decay rate; local density of optical states (LDOS); mislocalization; OPTICAL RECONSTRUCTION MICROSCOPY; LIGHT-MATTER INTERACTIONS; LOCAL-DENSITY; DIFFRACTION; ACCURACY; ANTENNA; STATES;
D O I
10.1021/acsphotonics.9b01317
中图分类号
TB3 [工程材料学];
学科分类号
0805 ; 080502 ;
摘要
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.
引用
收藏
页码:393 / 400
页数:15
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