Nanoplasmonics-enhanced label-free imaging of endothelial cell monolayer integrity

Frederic Banville (1, 2) , Julien Moreau (3) , Kevin Chabot (1, 2) , Andrea Cattoni (4) , Ulrike Fröhlich (5) , Jean-François Bryche (1, 2) , Stéphane Collin (4) , Paul Charette (1, 2) , Michel Grandbois (1) , Michael Canva (1, 2)

1 LN2 - Laboratoire Nanotechnologies Nanosystèmes
2 3IT - Institut Interdisciplinaire d'Innovation Technologique [Sherbrooke]
3 Laboratoire Charles Fabry / Biophotonique
4 LPN - Laboratoire de photonique et de nanostructures
5 UdeS - Université de Sherbrooke

Surface plasmon resonance imaging (SPRI) is a powerful label-free imaging modality for the analysis of morphological dynamics in cell monolayers. However, classical plasmonic imaging systems have relatively poor spatial resolution along one axis due to the plasmon mode attenuation distance (tens of μm, typically), which significantly limits their ability to resolve subcellular structures. We address this limitation by adding an array of nanostructures onto the metal sensing surface (25 nm thick, 200 nm width, 400 nm period grating) to couple localized plasmons with propagating plasmons, thereby reducing attenuation length and commensurately increasing spatial imaging resolution, without significant loss of sensitivity or image contrast. In this work, experimental results obtained with both conventional unstructured and nanostructured gold film SPRI sensor chips show a clear gain in spatial resolution achieved with surface nanostructuring. The work demonstrates the ability of the nanostructured SPRI chips to resolve fine morphological detail (intercellular gaps) in experiments monitoring changes in endothelial cell monolayer integrity following the activation of the cell surface protease-activated receptor 1 (PAR1) by thrombin. In particular, the nanostructured chips reveal the persistence of small intercellular gaps (< 5 μm 2) well after apparent recovery of cell monolayer integrity as determined by conventional unstructured surface based SPRI. This new high spatial resolution plasmonic imaging technique uses low-cost and reusable patterned substrates and is likely to find applications in cell biology and pharmacology by allowing label-free quantification of minute cell morphological activities associated with receptor dependent intracellular signaling activity.

Mots clés: spatial resolution; cell-based biosensing; Label-free microscopy; cell signaling; endothelium; plasmonics; nanostructured surface

Domaines: Optique / photonique; Micro et nanotechnologies/Microélectronique

Frederic Banville, Julien Moreau, Kevin Chabot, Andrea Cattoni, Ulrike Fröhlich, et al.. Nanoplasmonics-enhanced label-free imaging of endothelial cell monolayer integrity. Biosensors and Bioelectronics, 2019, 141, pp.111478. ⟨10.1016/j.bios.2019.111478⟩. ⟨hal-02184086⟩