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A new technique for reversible permeabilization of live cells for intracellular delivery of quantum dots

机译:活细胞可逆通透性用于量子点细胞内递送的新技术

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摘要

A major challenge with the use of quantum dots (QDs) for cellular imaging and biomolecular delivery is the attainment of QDs freely dispersed inside the cells. Conventional methods such as endocytosis, lipids based delivery and electroporation are associated with delivery of QDs in vesicles and/or as aggregates that are not monodispersed. In this study, we demonstrate a new technique for reversible permeabilization of cells to enable the introduction of freely dispersed QDs within the cytoplasm. Our approach combines osmosis driven fluid transport into cells achieved by creating a hypotonic environment and reversible permeabilization using low concentrations of cell permeabilization agents like Saponin. Our results confirm that highly efficient endocytosis-free intracellular delivery of QDs can be accomplished using this method. The best results were obtained when the cells were treated with 50 μg ml~(-1) Saponin in a hypotonic buffer at a 3:2 physiological buffer:DI water ratio for 5 min at 4°C.
机译:使用量子点(QD)进行细胞成像和生物分子传递的主要挑战是获得自由分散在细胞内的QD。诸如内吞作用,基于脂质的递送和电穿孔的常规方法与QD在囊泡中的递送和/或作为未单分散的聚集体有关。在这项研究中,我们展示了一种可逆的细胞通透性新技术,能够在细胞质内引入自由分散的QD。我们的方法结合了渗透作用驱动的液体向细胞内的运输,方法是通过创建低渗环境和使用低浓度的细胞透化剂(如皂素)来实现可逆的透化作用。我们的结果证实,使用这种方法可以完成QDs的高效无内吞作用。当在低渗缓冲液中以3:2生理缓冲液:去离子水的比率在4°C下用50μgml〜(-1)皂苷处理细胞5分钟时,可获得最佳结果。

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