title: 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.
authors.affiliation: Departments of Chemistry and.
authors.forename: Hee-Sun
authors.initials: HS
authors.lastname: Han
authors.affiliation: Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Niemeyer
authors.affiliation: Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114.
authors.forename: Yuhui
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114.
authors.forename: Walid S
authors.initials: WS
authors.lastname: Kamoun
authors.affiliation: Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114 Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139; and.
authors.forename: John D
authors.identifier: http://orcid.org/0000-0002-9828-8203
authors.initials: JD
authors.lastname: Martin
authors.affiliation: Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114.
authors.forename: Jayeeta
authors.initials: J
authors.lastname: Bhaumik
authors.affiliation: Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114.
authors.forename: Yunching
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Roberge
authors.affiliation: Departments of Chemistry and.
authors.forename: Jian
authors.initials: J
authors.lastname: Cui
authors.affiliation: Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114.
authors.forename: Margaret R
authors.initials: MR
authors.lastname: Martin
authors.affiliation: Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114.
authors.forename: Dai
authors.initials: D
authors.lastname: Fukumura
authors.affiliation: Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114 jain@steele.mgh.harvard.edu mgb@mit.edu.
authors.forename: Rakesh K
authors.initials: RK
authors.lastname: Jain
authors.affiliation: Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114 jain@steele.mgh.harvard.edu mgb@mit.edu.
authors.forename: Moungi G
authors.initials: MG
authors.lastname: Bawendi
authors.affiliation: Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114.
authors.forename: Dan G
authors.initials: DG
authors.lastname: Duda
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D001672:Biocompatible Materials; D051379:Mice; D008822:Mice, Transgenic; D045663:Quantum Dots
chemical_list: D000906:Antibodies; D001672:Biocompatible Materials
keywords: in vivo cytometry; in vivo imaging; multiphoton microscopy; quantum dots; single cell imaging