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用于在活细胞中快速实时追踪溶酶体运动和监测三磷酸腺苷(ATP)的基于碳点(CD)的荧光探针。

Carbon dot (CD)-based fluorescent probes for rapid and real-time tracking lysosomes movement and ATP monitoring in live cell.

作者信息

Ni Chuyao, Zhou Zihao, Cao Jun, Pan Jiaqi, Li Chaorong, Zheng Yingying

机构信息

Zhejiang Key Laboratory of Quantum State Control and Optical Field Manipulation, Department of Physics, Zhejiang Sci-Tech University, Hangzhou, 310018, China.

出版信息

Mikrochim Acta. 2025 Sep 11;192(10):658. doi: 10.1007/s00604-025-07537-8.

Abstract

The detection of lysosomal dysfunction and abnormal adenosine triphosphate (ATP) levels is crucial due to their roles in various cellular disorders and tumor progression, respectively. In this paper, carbon dot (CD)-based fluorescent probes (CDs-1 and CDs-2) with rapid lysosomal targeting and real-time lysosomal monitoring capability were designed. Through a hydrothermal method, the different nitrogen doping contents of CDs-1 and CDs-2 were controlled by varying the ethylenediamine content while the lysosome-targeting ability derived from their neutral red precursor was preserved. CDs-1 and CDs-2 rapid and effectively targeted lysosomes in both H1975 and 4T1 cells, with Pearson's colocalization coefficients of 0.94 and 0.88 (CDs-1) and 0.84 and 0.84 (CDs-2), closely matching the staining patterns of commercial lysosomal markers. CDs-1 demonstrated high specificity and sensitivity for ATP detection, displaying a linear detection range of 4-44 μM with a detection limit as low as 2.86 μM. CDs-1 enables  monitoring of intracellular ATP level changes in the live cells under etoposide stimulation, while CDs-2 exhibited outstanding photostability and enabled real-time and long-term tracking of lysosomal movement. This study establishes CD-based fluorescent probes as valuable tools for investigating lysosomal function, with potential applications in cell biology research, disease mechanism studies, and drug screening platforms.

摘要

由于溶酶体功能障碍和三磷酸腺苷(ATP)水平异常分别在各种细胞紊乱和肿瘤进展中发挥作用,因此对它们的检测至关重要。在本文中,设计了具有快速溶酶体靶向和实时溶酶体监测能力的基于碳点(CD)的荧光探针(CDs-1和CDs-2)。通过水热法,通过改变乙二胺含量来控制CDs-1和CDs-2的不同氮掺杂含量,同时保留源自其中性红前体的溶酶体靶向能力。CDs-1和CDs-2在H1975和4T1细胞中均能快速有效地靶向溶酶体,Pearson共定位系数分别为0.94和0.88(CDs-1)以及0.84和0.84(CDs-2),与商业溶酶体标记物的染色模式密切匹配。CDs-1对ATP检测表现出高特异性和灵敏度,线性检测范围为4-44μM,检测限低至2.86μM。CDs-1能够监测依托泊苷刺激下活细胞内ATP水平的变化,而CDs-2表现出出色的光稳定性,能够实时和长期跟踪溶酶体的运动。本研究将基于CD的荧光探针确立为研究溶酶体功能的有价值工具,在细胞生物学研究、疾病机制研究和药物筛选平台中具有潜在应用。

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