Cargando…

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

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...

Descripción completa

Detalles Bibliográficos
Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6411894/
https://www.ncbi.nlm.nih.gov/pubmed/30858536
http://dx.doi.org/10.1038/s41598-019-40612-8
_version_ 1783402477669842944
author 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
author_facet 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
author_sort King, Damien
collection PubMed
description 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.
format Online
Article
Text
id pubmed-6411894
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-64118942019-03-13 Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform 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 Sci Rep Article 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. Nature Publishing Group UK 2019-03-11 /pmc/articles/PMC6411894/ /pubmed/30858536 http://dx.doi.org/10.1038/s41598-019-40612-8 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
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
Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform
title Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform
title_full Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform
title_fullStr Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform
title_full_unstemmed Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform
title_short Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform
title_sort label-free multi parameter optical interrogation of endothelial activation in single cells using a lab on a disc platform
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6411894/
https://www.ncbi.nlm.nih.gov/pubmed/30858536
http://dx.doi.org/10.1038/s41598-019-40612-8
work_keys_str_mv AT kingdamien labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT glynnmacdara labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT cindricsandra labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT kernandavid labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT oconnelltriona labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT hakimjavadiroya labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT kearneysinead labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT ackermanntobias labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT berbelxaviermunoz labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT lloberaandreu labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT simonsenulf labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT laursenbritte labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT redmondeileenm labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT cahillpaula labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform
AT ducreejens labelfreemultiparameteropticalinterrogationofendothelialactivationinsinglecellsusingalabonadiscplatform