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Quantum dot mediated imaging of atherosclerosis

机译:量子点介导的动脉粥样硬化成像

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摘要

The progression of atherosclerosis is associated with leukocyte infiltration within lesions. We describe a technique for the ex vivo imaging of cellular recruitment in atherogenesis which utilizes quantum dots (QD) to color-code different cell types within lesion areas. Spectrally distinct QD were coated with the cell-penetrating peptide maurocalcine to fluorescently-label immunomagnetically isolated monocyte/macrophages and T lymphocytes. QD-maurocalcine bioconjugates labeled both cell types with a high efficiency, preserved cell viability, and did not perturb native leukocyte function in cytokine release and endothelial adhesion assays. QD-labeled monocyte/macrophages and T lymphocytes were reinfused in an ApoE~(-/-) mouse model of atherosclerosis and age-matched controls and tracked for up to four weeks to investigate the incorporation of cells within aortic lesion areas, as determined by oil red O (ORO) and immunofluorescence ex vivo staining. QD-labeled cells were visible in atherosclerotic plaques within two days of injection, and the two cell types colocalized within areas of subsequent ORO staining. Our method for tracking leukocytes in lesions enables high signal-to-noise ratio imaging of multiple cell types and biomarkers simultaneously within the same specimen. It also has great utility in studies aimed at investigating the role of distinct circulating leukocyte subsets in plaque development and progression.
机译:动脉粥样硬化的进展与病变内的白细胞浸润有关。我们描述了一种用于动脉粥样硬化中细胞募集的离体成像技术,该技术利用量子点(QD)对病变区域内的不同细胞类型进行颜色编码。光谱独特的QD涂有穿透细胞的肽maurocalcine,以荧光标记免疫磁性分离的单核细胞/巨噬细胞和T淋巴细胞。 QD-maurocalcine生物共轭物可高效标记两种细胞类型,保留细胞活力,并且在细胞因子释放和内皮粘附试验中不会干扰天然白细胞的功能。将QD标记的单核细胞/巨噬细胞和T淋巴细胞重新注入动脉粥样硬化和年龄匹配的对照组的ApoE〜(-/-)小鼠模型中,并追踪长达四周以研究主动脉病变区域内细胞的掺入情况,具体方法如下:油红O(ORO)和离体免疫荧光染色。 QD标记的细胞在注射后的两天内在动脉粥样硬化斑块中可见,并且两种细胞类型共定位在随后的ORO染色区域内。我们跟踪病变中白细胞的方法可在同一样本中同时对多种细胞类型和生物标记物进行高信噪比成像。它在旨在研究不同循环白细胞亚群在斑块发育和进程中的作用的研究中也具有很大的实用性。

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