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一种基于二硫化钼纳米片和银纳米簇的纳米探针,用于细胞内三磷酸腺苷的成像和定量分析。

A nanoprobe based on molybdenum disulfide nanosheets and silver nanoclusters for imaging and quantification of intracellular adenosine triphosphate.

作者信息

Xu Yan, Kang Qi, Yang Bin, Chen Beibei, He Man, Hu Bin

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

Anal Chim Acta. 2020 Oct 16;1134:75-83. doi: 10.1016/j.aca.2020.08.011. Epub 2020 Aug 20.

Abstract

Adenosine triphosphate (ATP), as a high-energy phosphate compound that stores and releases energy in living cells, has an irreplaceable role in many physiological processes and maintenance of biological functions, and can be used as an indicator of many diseases. In this work, a composite nanoprobe, silver nanocluster (AgNC) @ molybdenum disulfide (MoS), was designed to achieve in situ fluorescence imaging and quantitative analysis of intracellular ATP in HeLa cells by fluorescence spectrometry and inductively coupled plasma mass spectrometry (ICP-MS). The probe was constructed based on the adsorption of DNA-AgNCs by MoS nanosheets, and the DNA-AgNCs were prepared with the ATP aptamer as a template, whose fluorescence was initially quenched by MoS. When the probe was incubated into the cells, intracellular ATP recognized the aptamer sequence and caused the DNA-AgNCs to fall off the MoS nanosheets, resulting in fluorescence recovery. Here, AgNCs not only acted as a fluorescence label for imaging, but also as an element tag for quantitative analysis of intracellular ATP with the detection of Ag by ICP-MS. The ATP in HeLa cells detected by this method was 24.6 ± 1.7 nmol L, which was in good agreement with the test result of the ATP test kit (20.4 ± 0.8 nmol L). The proposed method has potential application in medical clinical diagnosis and evaluation of the body's metabolic level via fluorescence imaging and ICP-MS detection of intracellular ATP.

摘要

三磷酸腺苷(ATP)作为一种在活细胞中储存和释放能量的高能磷酸化合物,在许多生理过程和生物功能维持中具有不可替代的作用,并且可作为多种疾病的指标。在本工作中,设计了一种复合纳米探针,即银纳米簇(AgNC)@二硫化钼(MoS),通过荧光光谱法和电感耦合等离子体质谱法(ICP-MS)实现对HeLa细胞内ATP的原位荧光成像和定量分析。该探针基于MoS纳米片对DNA-AgNCs的吸附构建而成,DNA-AgNCs是以ATP适配体为模板制备的,其荧光最初被MoS淬灭。当将探针孵育到细胞中时,细胞内的ATP识别适配体序列并导致DNA-AgNCs从MoS纳米片上脱落,从而使荧光恢复。在此,AgNCs不仅作为成像的荧光标记,还作为通过ICP-MS检测Ag对细胞内ATP进行定量分析的元素标签。用该方法检测的HeLa细胞中的ATP为24.6±1.7 nmol L,与ATP检测试剂盒的测试结果(20.4±0.8 nmol L)吻合良好。所提出的方法通过对细胞内ATP的荧光成像和ICP-MS检测,在医学临床诊断和人体代谢水平评估方面具有潜在应用价值。

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