Reduced background autofluorescence for cell imaging using nanodiamonds and lanthanide chelates

Nicole M. Cordina, Nima Sayyadi, Lindsay M. Parker, Arun Everest-Dass, Louise J. Brown, Nicolle H. Packer*

*Corresponding author for this work

Research output: Contribution to journalArticle

12 Citations (Scopus)
10 Downloads (Pure)

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.

Original languageEnglish
Article number4521
Pages (from-to)1-14
Number of pages14
JournalScientific Reports
Volume8
Issue number1
DOIs
Publication statusPublished - 14 Mar 2018

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