Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform

Damien King, MacDara D. Glynn, Sandra Cindric, David Kernan, Tríona O’Connell, Roya Hakimjavadi, Sinéad Kearney, Tobias Ackermann, Xavier Munoz Berbel, Andreu Llobera, Ulf Simonsen, Britt E. Laursen, Eileen M. Redmond, Paul A. Cahill, Jens Ducrée

    Research output: Contribution to journalArticleResearch

    1 Citation (Scopus)

    Abstract

    © 2019, The Author(s). Cellular activation and inflammation leading to endothelial dysfunction is associated with cardiovascular disease (CVD). We investigated whether a single cell label-free multi parameter optical interrogation system can detect endothelial cell and endothelial progenitor cell (EPC) activation in vitro and ex vivo, respectively. Cultured human endothelial cells were exposed to increasing concentrations of tumour necrosis factor alpha (TNF-α) or lipopolysaccharide (LPS) before endothelial activation was validated using fluorescence-activated cell sorting (FACS) analysis of inflammatory marker expression (PECAM-1, E-selectin and ICAM-1). A centrifugal microfluidic system and V-cup array was used to capture individual cells before optical measurement of light scattering, immunocytofluorescence, auto-fluorescence (AF) and cell morphology was determined. In vitro, TNF-α promoted specific changes to the refractive index and cell morphology of individual cells concomitant with enhanced photon activity of fluorescently labelled inflammatory markers and increased auto-fluorescence (AF) intensity at three different wavelengths, an effect blocked by inhibition of downstream signalling with Iκβ. Ex vivo, there was a significant increase in EPC number and AF intensity of individual EPCs from CVD patients concomitant with enhanced PECAM-1 expression when compared to normal controls. This novel label-free ‘lab on a disc’ (LoaD) platform can successfully detect endothelial activation in response to inflammatory stimuli in vitro and ex vivo.
    Original languageEnglish
    Article number4157
    Pages (from-to)4157
    Number of pages14
    JournalScientific Reports
    Volume9
    Issue number1
    DOIs
    Publication statusPublished - 11 Mar 2019

    Keywords

    • DYSFUNCTION
    • EXPRESSION
    • ICAM-1
    • INFLAMMATION
    • MICROSCOPY
    • NF-KAPPA-B
    • OXIDATIVE STRESS
    • PROTEINS
    • REFRACTIVE-INDEX
    • TNF-ALPHA

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