期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 112, 期 5, 页码 1350-1355出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1421632111
关键词
quantum dots; in vivo imaging; single cell imaging; in vivo cytometry; multiphoton microscopy
资金
- National Institute of Health [R01-CA126642, R01-CA115767, P01-CA080124, R01-CA096915, U54-CA151884, P41-EB015871-26A1, R21-CA139168, R01-CA159258]
- Department of Defense Breast Cancer Research Innovator Award [W81XWH-10-1-0016]
- American Cancer Society [RSG-11-073-01-346TBG]
- Susan G.Komen Foundation
- Samsung Scholarship
- Massachusetts Institutes of Technology Department of Chemistry Instrumentation Facility [CHE-980806, DBI-9729592]
- Institute for Soldier Nanotechnology [W911NF-07-D-0004]
- National Science Foundation-Materials Research Science and Engineering Centers Program [DMR-0117795]
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 (similar to 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 (Sca(1+) 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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