Surface Plasmon Resonance to Study Cell Signaling and GPCR Functional Selectivity in Live Cells

Bourassa, Philippe; Söllradl, Thomas; Maltais, Jean-Sébastien; Charette, Paul G.; Gendron, Louis; Grandbois, Michel

doi:10.1007/978-1-4939-2617-6_10

Cited by 1 publication

(1 citation statement)

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“…Most commonly used techniques employ fluorescent probes to specifically mark structures and molecular targets, as it is the case for total internal reflection fluorescence (TIRF) and surface plasmon enhanced fluorescence (SPEF) (Chabot et al, 2013;Goedhart and Gadella, 2009;He et al, 2009;Liebermann and Knoll, 2000;Ni et al, 2018). However, while being highly specific techniques, the use of labelling may impact the cells (Progatzky et al, 2013), which is not the case for non-invasive labelfree methods (Bourassa et al, 2015;Bousse, 1996;Chabot et al, 2009;Fang, 2011;Pancrazio et al, 1999). Cell activity has been successfully monitored using several label-free biosensing methods, in particular by electrochemical impedance spectroscopy (Giaever and Keese, 1993;McGuiness, 2007), resonant Waveguide Grating (Fang et al, 2006;Ferrie et al, 2010), surface plasmon resonance (Chabot et al, 2013(Chabot et al, , 2012(Chabot et al, , 2009Giebel et al, 1999;Maltais et al, 2012;Peterson et al, 2009;Wang et al, 2012) and metal-clad waveguides (Söllradl et al, 2018b(Söllradl et al, , 2018a(Söllradl et al, , 2017.…”

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confidence: 99%

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

Banville

Moreau

Chabot

et al. 2019

Biosensors and Bioelectronics

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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.

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