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 μm2 with a single-molecule localization precision of ∼ 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.
Relocating Single Molecules in Super-Resolved Fluorescence Lifetime Images near a Plasmonic Nanostructure
Gulinatti A.;Acconcia G.;
2020-01-01
Abstract
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 μm2 with a single-molecule localization precision of ∼ 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.File | Dimensione | Formato | |
---|---|---|---|
2020_ACS_RelocatingSingleMolecules.pdf
Accesso riservato
Descrizione: Paper
:
Publisher’s version
Dimensione
3.06 MB
Formato
Adobe PDF
|
3.06 MB | Adobe PDF | Visualizza/Apri |
11311-1149164_Gulinatti.pdf
accesso aperto
:
Post-Print (DRAFT o Author’s Accepted Manuscript-AAM)
Dimensione
1.07 MB
Formato
Adobe PDF
|
1.07 MB | Adobe PDF | Visualizza/Apri |
I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.