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通过荧光相关光谱法对荧光标记的糖金纳米粒子的细胞内动力学进行定量研究。

A quantitative study of the intracellular dynamics of fluorescently labelled glyco-gold nanoparticles via fluorescence correlation spectroscopy.

机构信息

Biosurfaces Unit, Laboratory 1, CIC biomaGUNE, Paseo Miramón 182 C, San Sebastián, 20009, Spain.

出版信息

Small. 2014 Jul 9;10(13):2602-10. doi: 10.1002/smll.201303604. Epub 2014 Mar 17.

Abstract

The dynamic behaviour of gold nanoparticles functionalised with glucose (Glc-Au NPs) has been studied here by means of fluorescence correlation spectroscopy (FCS). Meaningful data on the state of aggregation and dynamics of Glc-Au NPs fluorescently-labelled with HiLyte Fluor647 (Glc-Au-Hi NPs) in the intracellular environment were obtained. Moreover, the work presented here shows that FCS can be used to visualise the presence of single NPs or NP aggregates following uptake and to estimate, locally, NP concentrations within the cell. FCS measurements become possible after applying a "prebleaching" methodology, when the immobile NP fraction has been effectively removed and thus significant FCS data has been recorded. In this study, Glc-Au-Hi NPs have been incubated with HepG2 cells and their diffusion time in the intracellular environment has been measured and compared with their diffusion value in water and cell media.

摘要

通过荧光相关光谱(FCS)研究了葡萄糖功能化的金纳米粒子(Glc-Au NPs)的动态行为。通过使用 HiLyte Fluor647 标记的 Glc-Au NPs(Glc-Au-Hi NPs)在细胞内环境中获得了有关聚集状态和动态的有意义数据。此外,本文的工作表明,FCS 可用于可视化摄取后单个 NPs 或 NP 聚集体的存在,并局部估计细胞内的 NP 浓度。在应用“预漂白”方法后,可以进行 FCS 测量,此时已经有效地去除了不移动的 NP 部分,从而记录了有意义的 FCS 数据。在这项研究中,将 Glc-Au-Hi NPs 与 HepG2 细胞孵育,并测量了它们在细胞内环境中的扩散时间,并将其与在水中和细胞培养基中的扩散值进行了比较。

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