Reduced background autofluorescence for cell imaging using nanodiamonds and lanthanide chelates

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 met...

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Bibliographic Details
Published in:Scientific reports 2018-03, Vol.8 (1), p.4521-14, Article 4521
Main Authors: Cordina, Nicole M, Sayyadi, Nima, Parker, Lindsay M, Everest-Dass, Arun, Brown, Louise J, Packer, Nicolle H
Format: Article
Language:English
Subjects:
Binding sites
Chelates
E-selectin
Endothelial cells
Fluorophores
Glycoproteins
I.R. radiation
Inflammation
Ligands
Point defects
Polyethylene glycol
Stains & staining
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Summary: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.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-018-22702-1