Nitrogen-doped carbon dots as a ratiometric fluorescent probe for determination of the activity of acid phosphatase, for inhibitor screening, and for intracellular imaging
Nitrogen-doped carbon dots as a ratiometric fluorescent probe for determination of the activity of acid phosphatase, for inhibitor screening, and for intracellular imaging
复制标题
氮掺杂碳点作为比率荧光探针，用于测定酸性磷酸酶的活性、用于抑制剂筛选和细胞内成像
DOI：
10.1007/s00604-019-3600-9
复制
发表时间：
2019
影响因子：
5.7
通讯作者：
Lin Liqing
中科院分区：
文献类型：
--
作者：
Zhu Zhenmao;Lin Xiaoyun;Wu Lina;Zhao Chengfei;Li Shaoguang;Liu Ailin;Lin Xinhua;Lin Liqing
介绍了用荷叶汁和乙二胺的混合物水热法制备绿色荧光氮掺杂碳点（N-CDs）的方法。N-CDs具有均匀的粒径、良好的分散性和水溶性。在316 nm和366 nm光激发下，它们表现出双荧光，发射峰分别在415 nm和509 nm处。它们带正电荷，具有较低的细胞毒性。这使它们成为荧光测定和生物成像的绝佳选择。建立了测定酸性磷酸酶（ACP）活性的比例分析法。它是基于六偏磷酸钠（NaPO3）6对N-CDs荧光的聚集诱导猝灭（AIQ）。ACP对（NaPO3）6的酶解导致（NaPO3）6的分解和荧光的恢复。测量两个波长（415 nm和509 nm）的荧光比，可以广泛消除背景干扰和波动信号。该方法的检测限为0.43 U·L−1，检测范围为1 ~ 50 U·L−1ACP。它成功地应用于(a)加标血清样品中ACP的测定，(b) ACP抑制剂的筛选，(c) HePG2细胞中ACP的成像。图解：氮掺杂碳点（N-CDs）的合成及其在基于聚集诱导猝灭和酶解的比值荧光法测定酸性磷酸酶（ACP）中的应用。
The author describe a method for preparation of green fluorescent nitrogen-doped carbon dots (N-CDs) through hydrothermal treatment of a mixture of lotus leaf juice and ethylenediamine (EDA). The N-CDs have uniform size, good dispersibility and water solubility. Under 316 and 366 nm photoexcitation, they show dual fluorescence with emission peaks at 415 and 509 nm, respectively. They are positively charge and display low cytotoxicity. This makes them an excellent choice for fluorometric assays and for bioimaging. A ratiometric assay was developed for the determination of the activity of acid phosphatase (ACP). It is based on the aggregation- induced quenching (AIQ) of the fluorescence of the N-CDs by sodium hexametaphosphate (NaPO3)6. Enzymatic hydrolysis of (NaPO3)6by ACP leads to the disintegration of (NaPO3)6and to the restoration of fluorescence. The measurement of the ratio of fluorescence at two wavelengths (415 and 509 nm), background interference and fluctuating signals can be widely eliminated. The method works in the 1–50 U·L−1ACP activity range and has a detection limit of 0.43 U·L−1. It was successfully applied (a) to the determination of ACP in spiked serum samples, (b) to ACP inhibitor screening, and (c) to imaging of ACP in HePG2 cells.Graphical abstractSchematic presentation of the synthesis of nitrogen-doped carbon dots (N-CDs), and their application to the ratiometric fluorometric determination of acid phosphatase (ACP) based on the aggregation-induced quenching and enzymatic hydrolysis.