Broadband plasmonic nanoantennas for multi-color nanoscale dynamics in living cells

TL;DR

This paper introduces broadband aluminum nanoantennas for multicolor, high-resolution imaging of molecular interactions in living cell membranes, enabling simultaneous detection and analysis of multiple biomolecules at the nanoscale.

Contribution

The study demonstrates the use of broadband aluminum nanoantennas for multicolor, nanoscale imaging of molecular interactions in living cells, surpassing limitations of narrow resonance gold structures.

Findings

Resolved nanoscale transient receptor interactions (~60 nm)

Detected individual receptor bursts underneath the antenna

Differentiated molecular diffusion within nanoclusters

Abstract

Recently, the implementation of plasmonic nanoantennas has opened new possibilities to investigate the nanoscale dynamics of individual biomolecules in living cell. However, studies have yet been restricted to single molecular species as the narrow wavelength resonance of gold-based nanostructures precludes the simultaneous interrogation of different fluorescently labeled molecules. Here we exploited broadband aluminum-based nanoantennas carved at the apex of near-field probes to resolve nanoscale-dynamic molecular interactions on intact living cell membranes. Through multicolor excitation, we simultaneously recorded fluorescence fluctuations of dual-color labeled transmembrane receptors known to form nanoclusters in living cells. Fluorescence cross-correlation studies revealed transient interactions between individual receptors in regions of ~60 nm. Moreover, the high signal-to-background ratio provided by the antenna illumination allowed us to directly detect fluorescent bursts arising from the passage of individual receptors underneath the antenna. Remarkably, by reducing the illumination volume below the characteristic receptor nanocluster sizes, we resolved molecular diffusion within nanoclusters and distinguished it from nanocluster diffusion. Spatiotemporal characterization of transient interactions between molecules is crucial to understand how they communicate with each other to regulate cell function. Our work demonstrates the potential of broadband photonic antennas to study multi-molecular events and interactions in living cell membranes with unprecedented spatiotemporal resolution.