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Han, H.S.* ; Niemeyer, E.* ; Huang, Y.* ; Kamoun, W.S.* ; Martin, J.D.* ; Bhaumik, J.* ; Chen, Y.* ; Roberge, S.* ; Cui, J. ; Martin, M.R.* ; Fukumura, D.* ; Jain, R.K.* ; Bawendi, M.G.* ; Duda, D.G.*

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

Proc. Natl. Acad. Sci. U.S.A. 112, 1350-1355 (2015)

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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.

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