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In vitro imaging of cells using peptide-conjugated quantum dots

机译：使用肽 - 共轭量子点的细胞体外成像

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Efficient intracellular delivery of quantum dots (QDs) in living cells and elucidating the mechanism of the delivery are essential for advancing the applications of QDs to in vivo imaging and in vivo photodynamic therapy. Here, we demonstrate that clathrin-mediated endocytosis is the most dominant pathway for the delivery of peptide-conjugated QDs. We selected an insect neuropeptide, allatostatin (AST1), conjugated with CdSe-ZnS QDs, and investigated the delivery of the conjugate in living cells. We evaluated the contributions of clathrin-mediated endocytosis, receptor-mediated endocytosis, and charge-based cell penetration to the delivery of QD605-AST1 conjugates by flow cytometry and fluorescence video microscopy. The delivery was suppressed by ～ 57% in inhibiting phosphoinositide 3-kinase with wortmannin, which blocks the formation of clathrin-coated vesicles, and by ～ 45% in incubating the cells at 4°C . Also, we identified clathrin-mediated endocytosis by two-color experiment to find colocalization of QD560-labeled clathrin heavy-chain antibody and QD605-AST1. We further observed reduction of the galanin receptor-mediated delivery of QD605-AST1 by ～ 8% in blocking the cells with a galanin antagonist, and reduction of charge-based cell penetration delivery by ～30% in removing the positive charge from arginine in the peptide and suppressing the cell-surface negative charge from glycosaminoglycan.
机译：高效细胞内递送量子点（QDS）在活细胞中并阐明递送机制对于推进QDS在体内成像和体内光动力疗法中的应用至关重要。在这里，我们证明克拉辛介导的内吞作用是递送肽缀合的QD的最占优势途径。我们选择了昆虫神经肽，阿拉特抑素（AST1），与Cdse-ZnS QD缀合，并研究了活细胞中的缀合物的递送。通过流式细胞术和荧光视频显微镜评估了通过流式细胞术和荧光视频显微镜对克拉仑介导的内吞作用，受体介导的内吞作用，受体介导的内吞作用和基于电荷的细胞渗透到QD605-AST1缀合物的贡献。抑制磷酸膦酸碱基三激酶抑制磷酸亚胺，抑制磷酸肌键，其阻断克拉林涂层的囊泡的形成，递送〜45％，在4℃下孵育细胞。此外，我们通过双色实验鉴定了克拉仑介导的内吞作用，以找到QD560标记的克拉辛重链抗体和QD605-AST1的分层化。我们进一步观察到将QD605-AST1的环肽受体介导的QD605-AST1的递送减少〜8％在阻止细胞与甘司蛋白拮抗剂，并将电荷的细胞渗透递送降低〜30％在除去精氨酸中的正电荷时肽和抑制来自糖氨基氨基聚糖的细胞表面负电荷。

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