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Monolayer surface chemistry enables 2-colour single molecule localisation microscopy of adhesive ligands and adhesion proteins

Nanofabricated and nanopatterned surfaces have revealed the sensitivity of cell adhesion to nanoscale variations in the spacing of adhesive ligands such as the tripeptide arginine-glycine-aspartic acid (RGD). To date, surface characterisation and cell adhesion are often examined in two separate expe...

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Autores principales: Lu, Xun, Nicovich, Philip R., Zhao, Manchen, Nieves, Daniel J., Mollazade, Mahdie, Vivekchand, S. R. C., Gaus, Katharina, Gooding, J. Justin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6102261/
https://www.ncbi.nlm.nih.gov/pubmed/30127420
http://dx.doi.org/10.1038/s41467-018-05837-7
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author Lu, Xun
Nicovich, Philip R.
Zhao, Manchen
Nieves, Daniel J.
Mollazade, Mahdie
Vivekchand, S. R. C.
Gaus, Katharina
Gooding, J. Justin
author_facet Lu, Xun
Nicovich, Philip R.
Zhao, Manchen
Nieves, Daniel J.
Mollazade, Mahdie
Vivekchand, S. R. C.
Gaus, Katharina
Gooding, J. Justin
author_sort Lu, Xun
collection PubMed
description Nanofabricated and nanopatterned surfaces have revealed the sensitivity of cell adhesion to nanoscale variations in the spacing of adhesive ligands such as the tripeptide arginine-glycine-aspartic acid (RGD). To date, surface characterisation and cell adhesion are often examined in two separate experiments so that the localisation of ligands and adhesion proteins cannot be combined in the same image. Here we developed self-assembled monolayer chemistry for indium tin oxide (ITO) surfaces for single molecule localisation microscopy (SMLM). Cell adhesion and spreading were sensitive to average RGD spacing. At low average RGD spacing, a threshold exists of 0.8 RGD peptides per µm(2) that tether cells to the substratum but this does not enable formation of focal adhesions. These findings suggest that cells can sense and engage single adhesive ligands but ligand clustering is required for cell spreading. Thus, our data reveal subtle differences in adhesion biology that may be obscured in ensemble measurements.
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spelling pubmed-61022612018-08-22 Monolayer surface chemistry enables 2-colour single molecule localisation microscopy of adhesive ligands and adhesion proteins Lu, Xun Nicovich, Philip R. Zhao, Manchen Nieves, Daniel J. Mollazade, Mahdie Vivekchand, S. R. C. Gaus, Katharina Gooding, J. Justin Nat Commun Article Nanofabricated and nanopatterned surfaces have revealed the sensitivity of cell adhesion to nanoscale variations in the spacing of adhesive ligands such as the tripeptide arginine-glycine-aspartic acid (RGD). To date, surface characterisation and cell adhesion are often examined in two separate experiments so that the localisation of ligands and adhesion proteins cannot be combined in the same image. Here we developed self-assembled monolayer chemistry for indium tin oxide (ITO) surfaces for single molecule localisation microscopy (SMLM). Cell adhesion and spreading were sensitive to average RGD spacing. At low average RGD spacing, a threshold exists of 0.8 RGD peptides per µm(2) that tether cells to the substratum but this does not enable formation of focal adhesions. These findings suggest that cells can sense and engage single adhesive ligands but ligand clustering is required for cell spreading. Thus, our data reveal subtle differences in adhesion biology that may be obscured in ensemble measurements. Nature Publishing Group UK 2018-08-20 /pmc/articles/PMC6102261/ /pubmed/30127420 http://dx.doi.org/10.1038/s41467-018-05837-7 Text en © The Author(s) 2018 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
Lu, Xun
Nicovich, Philip R.
Zhao, Manchen
Nieves, Daniel J.
Mollazade, Mahdie
Vivekchand, S. R. C.
Gaus, Katharina
Gooding, J. Justin
Monolayer surface chemistry enables 2-colour single molecule localisation microscopy of adhesive ligands and adhesion proteins
title Monolayer surface chemistry enables 2-colour single molecule localisation microscopy of adhesive ligands and adhesion proteins
title_full Monolayer surface chemistry enables 2-colour single molecule localisation microscopy of adhesive ligands and adhesion proteins
title_fullStr Monolayer surface chemistry enables 2-colour single molecule localisation microscopy of adhesive ligands and adhesion proteins
title_full_unstemmed Monolayer surface chemistry enables 2-colour single molecule localisation microscopy of adhesive ligands and adhesion proteins
title_short Monolayer surface chemistry enables 2-colour single molecule localisation microscopy of adhesive ligands and adhesion proteins
title_sort monolayer surface chemistry enables 2-colour single molecule localisation microscopy of adhesive ligands and adhesion proteins
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6102261/
https://www.ncbi.nlm.nih.gov/pubmed/30127420
http://dx.doi.org/10.1038/s41467-018-05837-7
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