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In vitro imaging of cells using peptide-conjugated quantum dots

机译：使用肽缀合的量子点对细胞进行体外成像

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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：量子点（QDs）在活细胞中的有效胞内传递以及阐明其传递机理对于推进QD在体内成像和体内光动力疗法中的应用至关重要。在这里，我们证明网格蛋白介导的内吞作用是肽偶联QD传递的最主要途径。我们选择了一种昆虫神经肽，阿托伐他汀（AST1），与CdSe-ZnS QDs缀合，并研究了缀合物在活细胞中的传递。我们通过流式细胞术和荧光视频显微镜评估了网格蛋白介导的内吞作用，受体介导的内吞作用和基于电荷的细胞渗透对QD605-AST1偶联物的递送的贡献。在用渥曼青霉素抑制磷酸肌醇3-激酶中，其递送被抑制了-57％，这阻止了网格蛋白包被的囊泡的形成，并且在-4℃温育细胞时被-45％抑制了递送。此外，我们通过双色实验鉴定了网格蛋白介导的内吞作用，以发现QD560标记的网格蛋白重链抗体和QD605-AST1的共定位。我们进一步观察到，在用甘丙肽拮抗剂阻断细胞时，甘丙肽受体介导的QD605-AST1传递降低了-8％，而在去除精氨酸中的正电荷后，基于电荷的细胞渗透传递降低了-30％。肽和抑制糖胺聚糖产生的细胞表面负电荷。

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