Plasmon-enhanced fluorescence correlation spectroscopy for super-localized detection of nanoscale subcellular dynamics

Biosens Bioelectron. 2021 Jul 15:184:113219. doi: 10.1016/j.bios.2021.113219. Epub 2021 Apr 22.

Abstract

In this report, we investigate plasmon-enhanced imaging fluorescence correlation spectroscopy (p-FCS). p-FCS takes advantage of extreme light confinement by localization at nanogap-based plasmonic nanodimer arrays (PNAs) for enhanced signal-to-noise ratio (SNR) and improved precision by registration with surface plasmon microscopy images. Theoretical results corroborate the enhancement by PNAs in the far-field. Near-field scanning optical microscopy was used to confirm near-field localization experimentally. Experimental confirmation was also conducted with fluorescent nanobeads. The concept was further applied to studying the diffusion dynamics of lysosomes in HEK293T cells stimulated by phorbol 12-myristate 13-acetate treatment. It was found that lysosomes demonstrate stronger super-diffusive behavior with relatively weaker sub-diffusion after stimulation. SNR measured of p-FCS was improved by 9.77 times over conventional FCS. This report is expected to serve as the foundation for an enhanced analytical tool to explore subcellular dynamics.

Keywords: Fluorescence correlation spectroscopy; Lysosome; Nanogap; Plasmonic nanodimer arrays; Plasmonics; Super-localization.

MeSH terms

  • Biosensing Techniques*
  • Diffusion
  • HEK293 Cells
  • Humans
  • Microscopy, Fluorescence
  • Spectrometry, Fluorescence