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

• Published in: Scientific reports 2019-03, Vol.9 (1), p.4157-4157, Article 4157
• Main Authors: King, Damien, Glynn, MacDara, Cindric, Sandra, Kernan, David, O’Connell, Tríona, Hakimjavadi, Roya, Kearney, Sinéad, Ackermann, Tobias, Berbel, Xavier Munoz, Llobera, Andreu, Simonsen, Ulf, Laursen, Britt E., Redmond, Eileen M., Cahill, Paul A., Ducrée, Jens
• Format: Article
• Language: English
• Subjects: 13/106; 13/2; 13/31; 13/62; 13/95; 14/1; 14/10; 14/34; 14/63; 631/80/2373; 639/624/1111/55; 692/4019/592/75; Cardiovascular diseases; CD31 antigen; Cell activation; Cell Shape; Cytology; E-selectin; E-Selectin - genetics; E-Selectin - metabolism; Endothelial cells; Flow cytometry; Flow Cytometry - instrumentation; Flow Cytometry - methods; Fluorescence; Human Umbilical Vein Endothelial Cells - cytology; Human Umbilical Vein Endothelial Cells - drug effects; Human Umbilical Vein Endothelial Cells - metabolism; Humanities and Social Sciences; Humans; Inflammation; Intercellular adhesion molecule 1; Intercellular Adhesion Molecule-1 - genetics; Intercellular Adhesion Molecule-1 - metabolism; Light scattering; Lipopolysaccharides; Lipopolysaccharides - pharmacology; Microfluidics; Morphology; multidisciplinary; Platelet Endothelial Cell Adhesion Molecule-1 - genetics; Platelet Endothelial Cell Adhesion Molecule-1 - metabolism; Progenitor cells; Questioning; Science; Science (multidisciplinary); Tumor Necrosis Factor-alpha - pharmacology; Tumor necrosis factor-TNF; Tumor necrosis factor-α; Tumors
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-40612-8

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 .