荧光成像-电感耦合等离子体质谱关联定量分析细胞内铜纳米粒子

引用本文: 郑安琪, 郝亚男, 果婷婷, 舒杨, 王建华. 荧光成像-电感耦合等离子体质谱关联定量分析细胞内铜纳米粒子[J]. 分析化学, 2020, 48(10): 1359-1366. doi: 10.19756/j.issn.0253-3820.201270 shu

Citation: ZHENG An-Qi, HAO Ya-Nan, GUO Ting-Ting, SHU Yang, WANG Jian-Hua. Correlation of Fluorescence Imaging with Inductively Coupled Plasma-Mass Spectrometry for Quantification of Intracellular Copper Nanoparticles[J]. Chinese Journal of Analytical Chemistry, 2020, 48(10): 1359-1366. doi: 10.19756/j.issn.0253-3820.201270 shu

荧光成像-电感耦合等离子体质谱关联定量分析细胞内铜纳米粒子

东北大学理学院化学系, 沈阳 110819
基金项目:
本文系国家自然科学基金项目（Nos.21974018，21727811）和中央高校基本科研业务费（Nos.N2005015，N2005027）资助

摘要: 建立了对细胞摄入的铜纳米粒子（GuNPs）进行定量分析的荧光成像法。以近红外发光的谷胱甘肽-铜纳米粒子（GSH-CuNPs）与MCF-7细胞孵育，利用共聚焦显微镜在激发波长559 nm、发射波长600~700 nm条件下对MCF-7细胞内的GSH-CuNPs进行荧光成像。细胞经裂解、离心超滤后，收集GSH-CuNPs，采用电感耦合等离子体质谱（ICP-MS）对铜元素进行定量分析。通过SPSS统计分析关联胞内平均荧光强度与ICP-MS的定量检测数据，建立平均荧光强度与细胞中CuNPs的定量关系；再以ICP-MS测定细胞内铜元素的总含量，可获得由GSH-CuNPs转化为其它形式的铜含量。采用本方法研究了MCF-7细胞中GSH-CuNPs的代谢/转化过程。结果表明，GSH-CuNPs与细胞孵育50 min后，细胞内CuNPs含量由172.74 fg/cell降低至18.66 fg/cell，且GSH-CuNPs主要代谢转化为可溶性铜。

关键词:

English

Correlation of Fluorescence Imaging with Inductively Coupled Plasma-Mass Spectrometry for Quantification of Intracellular Copper Nanoparticles

Department of Chemistry, College of Sciences, Northeastern University, Shenyang 110819, China
Received Date: 13 May 2020
Revised Date: 21 July 2020
Fund Project:
This work was supported by the National Natural Science Foundation of China (Nos. 21974018, 21727811) and the Fundamental Research Funds for the Central Universities (Nos. N2005015, N2005027).

Abstract: A fluorescence imaging protocol was developed for the quantification analysis of intracellular glutathione-copper nanoparticles (GSH-CuNPs). After culturing MCF-7 cells with near infrared luminescent GSH-CuNPs, the fluorescence imaging of intracellular CuNPs was achieved at λ_ex/λ_em=559/(600-700) nm. The cells were then subjected to lysis and ultrafiltration to collect the GSH-CuNPs. The copper content in the collected GSH-CuNPs was quantified by inductively coupled plasma-mass spectrometry (ICP-MS). Thereafter, the fluorescence density in the imaging was correlated with the analytical data by ICP-MS with SPSS statistics analysis to develop a linear calibration between the fluorescence intensity and the intracellular content of GSH-CuNPs. The developed protocol here was applied to the investigation of GSH-CuNPs metabolism/transformation in live MCF-7 cells, and it was found that with incubation time of 50 min, the content of GSH-CuNPs decreased from 172.74 fg/cell to 18.66 fg/cell and the majority of GSH-CuNPs were metabolized and transformed into soluble copper species.

Key words:

Copper nanoparticles
/ Cell
/ Fluorescence imaging
/ Quantification
/ Inductively coupled plasma-mass spectrometry

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

荧光成像-电感耦合等离子体质谱关联定量分析细胞内铜纳米粒子

English

Correlation of Fluorescence Imaging with Inductively Coupled Plasma-Mass Spectrometry for Quantification of Intracellular Copper Nanoparticles

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

荧光成像-电感耦合等离子体质谱关联定量分析细胞内铜纳米粒子

English

Correlation of Fluorescence Imaging with Inductively Coupled Plasma-Mass Spectrometry for Quantification of Intracellular Copper Nanoparticles

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

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CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

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