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William Carson

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Proceedings Article | 15 March 2016 Paper

Fluorescent nanodiamonds and their use in biomedical research

Lorena Suarez-Kelly, Isaac Rampersaud, Charles Moritz, Amanda Campbell, Zhiwei Hu, Masfer Alkahtani, Fahad Alghannam, Phillip Hemmer, William Carson, Arfaan Rampersaud

Proceedings Volume 9762, 976205 (2016) https://doi.org/10.1117/12.2218612

KEYWORDS: Diamond, Luminescence, Medical research, Microscopy, Flow cytometry, Crystals, Nitrogen, Chemical species, Carbon, Near infrared, Molecules, Confocal microscopy, Visualization, Biomedical optics

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Nanodiamonds containing color-centers produce non-quenching fluorescence that is easily detected. This makes them useful for cellular, proteomic and genomic applications. However, fluorescent nanodiamonds have yet to become popular in the biomedical research community as labeling reagents. We discuss production of nanodiamonds with distinct color-centers and assess their biocompatibility and techniques for bioconjugation. Fluorescent diamonds were fabricated by electron irradiation of high-pressure, high-temperature micron-sized diamonds which generated diamonds with vacancy-related defects (V). These diamonds were annealed to create nitrogen vacancy (NV)-centers then following a milling step were fractionated into nanoparticle sizes of 30, 60, and 95 nm. Optical characterization of Vand NV-center diamonds demonstrated fluorescence in two distinct green and red channels, respectively. In vitro studies demonstrated that these nanodiamonds are biocompatible and readily taken up by murine macrophage cells. Quantification of NV-center nanodiamond uptake by flow cytometry, showed that uptake was independent of nanodiamond size. Confocal microscopy demonstrated that NV-center nanodiamonds accumulate within the cytoplasm of these cells. NV-center nanodiamonds were then conjugated with streptavidin using a short polyethylene chain as linker. Conjugation was confirmed via a catalytic assay employing biotinylated-horseradish peroxidase. We present a technique for large-scale production of biocompatible conjugated V- or NV-center nanodiamonds. Functional testing is essential for standardization of fluorescent nanodiamond bioconjugates and quality control. Large-scale production of bioconjugated fluorescent nanodiamonds is crucial to their development as novel tools for biological and medical applications.

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