title: Wide-field imaging and flow cytometric analysis of cancer cells in blood by fluorescent nanodiamond labeling and time gating.
abstract: Nanodiamonds containing high density ensembles of negatively charged nitrogen-vacancy (NV(-)) centers are promising fluorescent biomarkers due to their excellent photostability and biocompatibility. The NV(-) centers in the particles have a fluorescence lifetime of up to 20 ns, which distinctly differs from those (<10 ns) of cell and tissue autofluorescence, making it possible to achieve background-free detection in vivo by time gating. Here, we demonstrate the feasibility of using fluorescent nanodiamonds (FNDs) as optical labels for wide-field time-gated fluorescence imaging and flow cytometric analysis of cancer cells with a nanosecond intensified charge-coupled device (ICCD) as the detector. The combined technique has allowed us to acquire fluorescence images of FND-labeled HeLa cells in whole blood covered with a chicken breast of ~0.1-mm thickness at the single cell level, and to detect individual FND-labeled HeLa cells in blood flowing through a microfluidic device at a frame rate of 23 Hz, as well as to locate and trace FND-labeled lung cancer cells in the blood vessels of a mouse ear. It opens a new window for real-time imaging and tracking of transplanted cells (such as stem cells) in vivo.
authors.affiliation: 1] Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 106, Taiwan [2].
authors.forename: Yuen Yung
authors.initials: YY
authors.lastname: Hui
authors.affiliation: 1] Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 106, Taiwan [2] Department of Chemistry, National Taiwan University, Taipei 106, Taiwan [3].
authors.forename: Long-Jyun
authors.initials: LJ
authors.lastname: Su
authors.affiliation: Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 106, Taiwan.
authors.forename: Oliver Yenjyh
authors.initials: OY
authors.lastname: Chen
authors.affiliation: Department of Chemistry, National Taiwan University, Taipei 106, Taiwan.
authors.forename: Yit-Tsong
authors.initials: YT
authors.lastname: Chen
authors.affiliation: Institute of Biomedical Engineering, National Taiwan University, Taipei 100, Taiwan.
authors.forename: Tzu-Ming
authors.initials: TM
authors.lastname: Liu
authors.affiliation: Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 106, Taiwan.
authors.forename: Huan-Cheng
authors.initials: HC
authors.lastname: Chang
mesh_terms: D000818:Animals; D058948:Cell Tracking; D002645:Chickens; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008807:Mice, Inbred BALB C; D046210:Microfluidic Analytical Techniques; D008856:Microscopy, Fluorescence; D058612:Nanodiamonds / Q000737:chemistry*; D009368:Neoplasm Transplantation; D009360:Neoplastic Cells, Circulating / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D058612:Nanodiamonds