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利用共聚焦激光扫描显微镜与激光烧蚀 ICPMS 联用对 HepG2 细胞中的铜转运蛋白 1 进行双模成像。

Dual-mode imaging of copper transporter 1 in HepG2 cells by hyphenating confocal laser scanning microscopy with laser ablation ICPMS.

机构信息

Research Center for Analytical Sciences, Department of Chemistry, College of Sciences, Northeastern University, Shenyang, 110819, Liaoning, China.

出版信息

Anal Bioanal Chem. 2021 Feb;413(5):1353-1361. doi: 10.1007/s00216-020-03097-3. Epub 2021 Jan 6.

Abstract

Copper transporter 1 (CTR1) is a transport protein involved in copper and cisplatin uptake. The visualization of cellular CTR1 migration and its redistribution is highly important in copper/cisplatin exposure/transport. However, to the best of our knowledge, this is a highly challenging task. Herein, a dual-mode imaging strategy for CTR1 is developed by hyphenating confocal laser scanning microscopy (CLSM) and laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS) with a fluorescent/elemental bifunctional tag conjugated with anti-CTR1 antibody. The tag consists of rhodamine B and zirconium metal-organic frameworks (Zr-MOF) for CLSM fluorescence imaging and LA-ICPMS element imaging for a same group of HepG2 cells in a designated visual zone. This dual-mode imaging strategy facilitates visualization of CTR1 migration and meanwhile provides information of CTR1 redistribution in HepG2 cells by uptake of divalent copper or cisplatin. The present dual-mode imaging strategy provides in-depth information for the elucidation of CTR1 involved biological processes. Graphical abstract.

摘要

铜转运蛋白 1(CTR1)是一种参与铜和顺铂摄取的转运蛋白。细胞内 CTR1 迁移及其重分布的可视化在铜/顺铂暴露/转运中非常重要。然而,据我们所知,这是一项极具挑战性的任务。在此,通过将共聚焦激光扫描显微镜(CLSM)和激光烧蚀电感耦合等离子体质谱(LA-ICPMS)与荧光/元素双功能标签偶联抗 CTR1 抗体相连接,开发了一种用于 CTR1 的双模成像策略。该标签由罗丹明 B 和锆金属-有机骨架(Zr-MOF)组成,用于 CLSM 荧光成像,以及 LA-ICPMS 元素成像,用于在指定的可视区域对同一组 HepG2 细胞进行成像。这种双模成像策略不仅有助于可视化 CTR1 的迁移,还通过摄取二价铜或顺铂为 HepG2 细胞中 CTR1 的再分布提供了信息。本双模成像策略为阐明 CTR1 参与的生物学过程提供了深入的信息。图摘要。

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