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Quantum dot/antibody conjugates for in vivo cytometric imaging in mice.


ABSTRACT: Multiplexed, phenotypic, intravital cytometric imaging requires novel fluorophore conjugates that have an appropriate size for long circulation and diffusion and show virtually no nonspecific binding to cells/serum while binding to cells of interest with high specificity. In addition, these conjugates must be stable and maintain a high quantum yield in the in vivo environments. Here, we show that this can be achieved using compact (?15 nm in hydrodynamic diameter) and biocompatible quantum dot (QD) -Ab conjugates. We developed these conjugates by coupling whole mAbs to QDs coated with norbornene-displaying polyimidazole ligands using tetrazine-norbornene cycloaddition. Our QD immunoconstructs were used for in vivo single-cell labeling in bone marrow. The intravital imaging studies using a chronic calvarial bone window showed that our QD-Ab conjugates diffuse into the entire bone marrow and efficiently label single cells belonging to rare populations of hematopoietic stem and progenitor cells (Sca1(+)c-Kit(+) cells). This in vivo cytometric technique may be useful in a wide range of structural and functional imaging to study the interactions between cells and between a cell and its environment in intact and diseased tissues.

SUBMITTER: Han HS 

PROVIDER: S-EPMC4321304 | biostudies-literature | 2015 Feb

REPOSITORIES: biostudies-literature

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Quantum dot/antibody conjugates for in vivo cytometric imaging in mice.

Han Hee-Sun HS   Niemeyer Elisabeth E   Huang Yuhui Y   Kamoun Walid S WS   Martin John D JD   Bhaumik Jayeeta J   Chen Yunching Y   Roberge Sylvie S   Cui Jian J   Martin Margaret R MR   Fukumura Dai D   Jain Rakesh K RK   Bawendi Moungi G MG   Duda Dan G DG  

Proceedings of the National Academy of Sciences of the United States of America 20150120 5


Multiplexed, phenotypic, intravital cytometric imaging requires novel fluorophore conjugates that have an appropriate size for long circulation and diffusion and show virtually no nonspecific binding to cells/serum while binding to cells of interest with high specificity. In addition, these conjugates must be stable and maintain a high quantum yield in the in vivo environments. Here, we show that this can be achieved using compact (∼15 nm in hydrodynamic diameter) and biocompatible quantum dot (  ...[more]

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