Reduced background autofluorescence for cell imaging using nanodiamonds and lanthanide chelates

Cordina, NM; Sayyadi, N; Parker, LM; Everest-Dass, A; Brown, LJ; Packer, NH

Reduced background autofluorescence for cell imaging using nanodiamonds and lanthanide chelates

Cordina, NM Sayyadi, N

Parker, LM Everest-Dass, A Brown, LJ Packer, NH

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Publication Type:: Journal Article
Citation:: Scientific Reports, 2018, 8 (1)
Issue Date:: 2018-12-01

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Field	Value	Language
dc.contributor.author	Cordina, NM	en_US
dc.contributor.author	Sayyadi, N https://orcid.org/0000-0002-0044-2170	en_US
dc.contributor.author	Parker, LM	en_US
dc.contributor.author	Everest-Dass, A	en_US
dc.contributor.author	Brown, LJ	en_US
dc.contributor.author	Packer, NH	en_US
dc.date.available	2018-02-22	en_US
dc.date.issued	2018-12-01	en_US
dc.identifier.citation	Scientific Reports, 2018, 8 (1)	en_US
dc.identifier.uri	http://hdl.handle.net/10453/137858
dc.description.abstract	© 2018 The Author(s). Bio-imaging is a key technique in tracking and monitoring important biological processes and fundamental biomolecular interactions, however the interference of background autofluorescence with targeted fluorophores is problematic for many bio-imaging applications. This study reports on two novel methods for reducing interference with cellular autofluorescence for bio-imaging. The first method uses fluorescent nanodiamonds (FNDs), containing nitrogen vacancy centers. FNDs emit at near-infrared wavelengths typically higher than most cellular autofluorescence; and when appropriately functionalized, can be used for background-free imaging of targeted biomolecules. The second method uses europium-chelating tags with long fluorescence lifetimes. These europium-chelating tags enhance background-free imaging due to the short fluorescent lifetimes of cellular autofluorescence. In this study, we used both methods to target E-selectin, a transmembrane glycoprotein that is activated by inflammation, to demonstrate background-free fluorescent staining in fixed endothelial cells. Our findings indicate that both FND and Europium based staining can improve fluorescent bio-imaging capabilities by reducing competition with cellular autofluorescence. 30 nm nanodiamonds coated with the E-selectin antibody was found to enable the most sensitive detective of E-selectin in inflamed cells, with a 40-fold increase in intensity detected.	en_US
dc.relation.ispartof	Scientific Reports	en_US
dc.relation.isbasedon	10.1038/s41598-018-22702-1	en_US
dc.rights	info:eu-repo/semantics/openAccess
dc.subject.mesh	Lanthanoid Series Elements	en_US
dc.subject.mesh	Chelating Agents	en_US
dc.subject.mesh	Fluorescent Dyes	en_US
dc.subject.mesh	Microscopy, Fluorescence	en_US
dc.subject.mesh	Protein Binding	en_US
dc.subject.mesh	Fluorescence	en_US
dc.subject.mesh	Molecular Imaging	en_US
dc.subject.mesh	Nanodiamonds	en_US
dc.subject.mesh	Signal-To-Noise Ratio	en_US
dc.subject.mesh	Biomarkers	en_US
dc.title	Reduced background autofluorescence for cell imaging using nanodiamonds and lanthanide chelates	en_US
dc.type	Journal Article
utslib.citation.volume	1	en_US
utslib.citation.volume	8	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology/School of Biomedical Engineering
utslib.copyright.status	open_access	*
pubs.issue	1	en_US
pubs.publication-status	Published	en_US
pubs.volume	8	en_US

Abstract:

© 2018 The Author(s). Bio-imaging is a key technique in tracking and monitoring important biological processes and fundamental biomolecular interactions, however the interference of background autofluorescence with targeted fluorophores is problematic for many bio-imaging applications. This study reports on two novel methods for reducing interference with cellular autofluorescence for bio-imaging. The first method uses fluorescent nanodiamonds (FNDs), containing nitrogen vacancy centers. FNDs emit at near-infrared wavelengths typically higher than most cellular autofluorescence; and when appropriately functionalized, can be used for background-free imaging of targeted biomolecules. The second method uses europium-chelating tags with long fluorescence lifetimes. These europium-chelating tags enhance background-free imaging due to the short fluorescent lifetimes of cellular autofluorescence. In this study, we used both methods to target E-selectin, a transmembrane glycoprotein that is activated by inflammation, to demonstrate background-free fluorescent staining in fixed endothelial cells. Our findings indicate that both FND and Europium based staining can improve fluorescent bio-imaging capabilities by reducing competition with cellular autofluorescence. 30 nm nanodiamonds coated with the E-selectin antibody was found to enable the most sensitive detective of E-selectin in inflamed cells, with a 40-fold increase in intensity detected.

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http://hdl.handle.net/10453/137858