College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Institute of Molecular and Nano Science, Shandong Normal University , Jinan, Shandong 250014, P. R. China.
Anal Chem. 2017 Apr 18;89(8):4559-4565. doi: 10.1021/acs.analchem.6b05045. Epub 2017 Apr 4.
Various intracellular metal ions have closely related functional roles in the nervous system. An excess or deficiency of essential metal ions can contribute to neurodegenerative diseases. Thus, the detection of various metal ions in neurons is important for diagnosing and monitoring these diseases. In particular, single-cell analysis of multiple metal ions allows us to not only understand the cellular heterogeneity and differentiation but also determine the actual relationships among multiple metal ions in each individual cell. Aiming at the low efficient single-cell manipulation and interference of complex biological matrices within cells in the existing method for single-cell metal ion detection, in this manuscript, we present a convenient, sensitive, and reliable method to simultaneously identify and quantify multiple metal ions at the single-cell level using a microfluidic system. Using the combination of on-chip electrophoresis separation and multicolor fluorescence detection, we achieved the simultaneous analysis of Na, K, Ca, and Mg in single PC-12 cells and studied changes in these four metal ions in Aβ-treated PC-12 cells, which is a model of Alzheimer's disease (AD). The data showed that metal ions imbalances in neuron-like cells may be associated with AD induced by Aβ. This method paves the way for multiple metal ion detection in single neuron-like cells, and the results provide insights regarding synergistic function of multiple metal ions in regulation of neurological diseases at the single-cell level.
各种细胞内金属离子在神经系统中具有密切相关的功能作用。必需金属离子的过量或缺乏都可能导致神经退行性疾病。因此，检测神经元中的各种金属离子对于诊断和监测这些疾病非常重要。特别是，对多种金属离子进行单细胞分析不仅可以帮助我们了解细胞的异质性和分化，还可以确定每个细胞内多种金属离子之间的实际关系。针对现有单细胞金属离子检测方法中存在的细胞内复杂生物基质对单细胞操作和干扰效率低的问题，本研究提出了一种使用微流控系统在单细胞水平上同时识别和定量多种金属离子的简便、灵敏、可靠的方法。我们结合片上电泳分离和多色荧光检测，实现了对单个 PC-12 细胞中 Na、K、Ca 和 Mg 的同时分析，并研究了 Aβ 处理的 PC-12 细胞中这四种金属离子的变化，这是阿尔茨海默病 (AD) 的模型。研究数据表明，神经元样细胞中的金属离子失衡可能与 Aβ 诱导的 AD 有关。该方法为单细胞中多种金属离子的检测铺平了道路，结果为单细胞水平上调节神经疾病的多种金属离子的协同作用提供了新的见解。
