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利用基于 CD 的比率荧光纳米探针实时监测活细胞内源性半胱氨酸水平。

Real-time monitoring of endogenous cysteine levels in living cells using a CD-based ratiometric fluorescent nanoprobe.

机构信息

Key Laboratory of Theoretical Organic Chemistry and Functional Molecule of Ministry of Education, Hunan Provincial Key Laboratory of Controllable Preparation and Functional Application of Fine Polymers, Hunan Province College Key Laboratory of QSAR/QSPR, Institute of Functional Materials, School of Chemistry and Chemical Engineering, Hunan University of Science and Technology, Xiangtan, 411201, Hunan, China.

State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha, 410082, China.

出版信息

Anal Bioanal Chem. 2018 Jul;410(18):4379-4386. doi: 10.1007/s00216-018-1091-x. Epub 2018 Apr 30.

Abstract

A simple and readily available fluorescent probe is needed for the real-time monitoring of endogenous cysteine (Cys) levels in living cells, as such a probe could be used to study the role of Cys in related diseases. Herein, we report the first fluorescent probe based on carbon dots (CDs-FITA) for the selective and ratiometric imaging of endogenous Cys in live cells. In this ratiometric fluorescent probe, a fluorescein derivative (FITA) that recognizes Cys is covalently linked to the surfaces of carbon dots (CDs); employing CDs greatly improves the water solubility of the probe. Acrylate on FITA is selectively cleaved by Cys in aqueous solution under mild conditions, leading to a dramatic increase in the fluorescence from fluorescein. The probe therefore allows the highly selective ratiometric fluorescent detection of Cys even in the presence of various interferents. The as-prepared CDs-FITA showed excellent performance when applied to detect Cys in blood serum. In addition, due to its negligible cytotoxicity, the CDs-FITA can also be utilized for the real-time monitoring of endogenous cysteine (Cys) levels in living cells. Graphical abstract Illustration of the CD-based probe for Cys imaging in living cells.

摘要

需要一种简单且易于获得的荧光探针来实时监测活细胞内内源性半胱氨酸(Cys)的水平,因为这种探针可用于研究 Cys 在相关疾病中的作用。在此,我们报告了第一个基于碳点(CDs-FITA)的荧光探针,用于活细胞中内源性 Cys 的选择性和比率成像。在这个比率荧光探针中,识别 Cys 的荧光素衍生物(FITA)通过共价键连接到碳点(CDs)的表面;使用 CDs 极大地提高了探针的水溶性。在水溶液中,FITA 上的丙烯酰胺在温和条件下被 Cys 选择性切割,导致荧光素的荧光显著增强。因此,该探针即使在存在各种干扰物的情况下,也能够高度选择性地进行比率荧光检测 Cys。所制备的 CDs-FITA 在用于检测血清中的 Cys 时表现出优异的性能。此外,由于其细胞毒性可以忽略不计,因此 CDs-FITA 还可以用于实时监测活细胞内内源性半胱氨酸(Cys)的水平。

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