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使用液滴微流控芯片-电感耦合等离子体质谱法研究金纳米粒子被单细胞摄取的情况。

Study on uptake of gold nanoparticles by single cells using droplet microfluidic chip-inductively coupled plasma mass spectrometry.

机构信息

Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, China.

Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, China.

出版信息

Talanta. 2019 Aug 1;200:398-407. doi: 10.1016/j.talanta.2019.03.075. Epub 2019 Mar 20.

DOI:10.1016/j.talanta.2019.03.075
PMID:31036201
Abstract

Cellular uptake of Au NPs is the premise for their application in drug delivery, cell/tissue imaging and quantitation analysis of biomarkers in the cells. However, conventional methods for cell analysis may lead to the loss of important information from abnormal or atypical cells in the cell population. In this work, with 15, 30 and 60 nm Au NPs@citric acid and Au NPs@DNA as model materials, the cellular uptake of Au NPs in single HeLa cells is studied by using an on-line droplet chip inductively coupled plasma mass spectrometry hyphenated technique. It is found that the average amount of Au uptake for Au NPs@citric acid is higher than that for Au NPs@DNA at high incubation concentrations. The distribution of the number of uptaken Au NPs in single HeLa cells shows a great difference when HeLa cells are incubated with different Au NPs. The percentage of cells uptaking Au NPs in the cell population also reveals a difference in cellular uptake between Au NPs@citric acid and Au NPs@DNA at the single-cell level. To explain the abovementioned phenomenon, the endocytosis mechanisms of Au NPs are investigated. Clathrin-mediated endocytosis is found to be the major internalization pathway for 15 and 30 nm Au NPs@DNA.

摘要

细胞内摄取 Au NPs 是其在药物输送、细胞/组织成像和细胞内生物标志物定量分析中应用的前提。然而,传统的细胞分析方法可能导致细胞群体中异常或非典型细胞的重要信息丢失。在这项工作中,以 15、30 和 60nm Au NPs@citric acid 和 Au NPs@DNA 为模型材料,使用在线液滴芯片电感耦合等离子体质谱联用技术研究了 Au NPs 在单个 HeLa 细胞中的摄取。结果发现,在高孵育浓度下,Au NPs@citric acid 的 Au 摄取平均量高于 Au NPs@DNA。当 HeLa 细胞用不同的 Au NPs 孵育时,单个 HeLa 细胞中摄取的 Au NPs 数量的分布有很大的差异。细胞群体中摄取 Au NPs 的细胞百分比也揭示了在单细胞水平上,Au NPs@citric acid 和 Au NPs@DNA 之间的细胞摄取存在差异。为了解释上述现象,研究了 Au NPs 的内吞作用机制。发现网格蛋白介导的内吞作用是 15 和 30nm Au NPs@DNA 的主要内化途径。

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