Reduced background autofluorescence for cell imaging using nanodiamonds and lanthanide chelates

Research output: Contribution to journalArticleResearchpeer-review

Abstract

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.

LanguageEnglish
Article number4521
Pages1-14
Number of pages14
JournalScientific Reports
Volume8
Issue number1
DOIs
Publication statusPublished - 14 Mar 2018

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Nanodiamonds
Lanthanoid Series Elements
Optical Imaging
Europium
E-Selectin
Staining and Labeling
Biological Phenomena
Glycoproteins
Nitrogen
Endothelial Cells
Fluorescence
Inflammation
Antibodies

Cite this

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title = "Reduced background autofluorescence for cell imaging using nanodiamonds and lanthanide chelates",
abstract = "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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Reduced background autofluorescence for cell imaging using nanodiamonds and lanthanide chelates. / Cordina, Nicole M.; Sayyadi, Nima; Parker, Lindsay M.; Everest-Dass, Arun; Brown, Louise J.; Packer, Nicolle H.

In: Scientific Reports, Vol. 8, No. 1, 4521, 14.03.2018, p. 1-14.

Research output: Contribution to journalArticleResearchpeer-review

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