研究目的
Investigating the spectral reshaping of single dye molecules coupled to single plasmonic nanoparticles to understand the light-matter interactions and the effects of local environment on fluorescence emission.
研究成果
Single-molecule hyperspectral imaging reveals that dyes with emission bluer than the LSPR experience a red-shifting of the emission spectrum, and dyes with emission redder than the LSPR experience a blue-shifting. The extent of spectral reshaping agrees with electromagnetic simulations, demonstrating the power of single-molecule hyperspectral imaging in studying light-matter interactions.
研究不足
The study does not adjust for mislocalization of molecules due to coupling with the nanoparticle, which affects the apparent position of a molecule. Differences between experimental results and simulation may also be attributed to particle heterogeneity.
1:Experimental Design and Method Selection:
Single-molecule hyperspectral super-resolution fluorescence imaging was used to measure the emission spectrum and super-resolved position of fluorophores near gold nanoparticles (AuNPs).
2:Sample Selection and Data Sources:
Glass coverslips decorated with AuNPs and four fluorescent dyes (BDP-FL, BDP-R6G, Cy3, Cy
3:5) with different spectral overlaps with the AuNP scattering spectrum were used. List of Experimental Equipment and Materials:
Inverted microscope (Olympus IX-71),
4:4 NA 100× oil-immersion objective (Olympus UPLSAPO 100XO), 488-nm laser (Coherent Sapphire), EMCCD cameras (Andor iXon 897, Andor iXon 887), spectrometer (Princeton Instruments SP-2300i). Experimental Procedures and Operational Workflow:
Dye solutions were introduced over the substrate, and single-molecule images and spectra were acquired upon stochastic adsorption of dye molecules onto the coverslip.
5:Data Analysis Methods:
Background subtraction using SMALL-LABS method, super-localization and spectral deconvolution with two-term Gaussian fitting.
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