Nano-imaging of trace metals by synchrotron X-ray fluorescence into dopaminergic single cells and neurite-like processes
Nano-imaging of trace metals by synchrotron X-ray fluorescence into dopaminergic single cells and neurite-like processes
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DOI：
10.1039/b802242a
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发表时间：
2008-01-01
影响因子：
3.4
通讯作者：
Ortega, Richard
中科院分区：
文献类型：
--
作者：
Carmona, Asuncion;Cloetens, Peter;Ortega, Richard
金属组被定义为金属和类金属在不同物种和细胞隔室中的分布。在亚细胞水平上检测微量元素是一项具有挑战性的任务，需要复杂的分析发展。在这项研究中，我们报告了如何使用最近在欧洲同步辐射设施开发的X射线荧光纳米探针在高空间分辨率下在多巴胺能细胞的亚细胞室中进行化学元素成像。高的空间分辨率是使用的概念，聚焦到一个90 nm的探头由多层反射镜弯曲在柯克帕特里克-贝兹几何的二次源。这种原始的设置被应用于单个多巴胺能细胞的痕量金属映射，选择作为参与帕金森病的退行性细胞的体外模型。这种细胞模型能够在暴露于神经生长因子时分化并延伸神经突样过程。获得了两个重要结果。首先，铁以颗粒形式分布到多巴胺囊泡中，主要存在于初级神经突生长和远端。第二，分化细胞产生的薄的神经突样突起积累铜、锌和少量铅。总的来说，单个神经元细胞的高分辨率成像提供了独特的信息，以了解微量金属在神经化学中的作用。
The metallome has been defined as the distribution of metals and metalloids among the different species and cell compartments. The detection of trace elements at the subcellular level is a challenging task that requires sophisticated analytical developments. In this study, we report how chemical element imaging was performed in subcellular compartments of dopaminergic cells at high spatial resolution using the X-ray fluorescence nanoprobe recently developed at the European Synchrotron Radiation Facility. High spatial resolution is obtained using the concept of a secondary source focused to a 90 nm probe by multilayer mirrors bent in Kirkpatrick - Baez geometry. This original setup was applied for trace metal mapping of single dopaminergic cells, chosen as an in vitro model of degenerative cells involved in Parkinson's disease. This cellular model is able to differentiate upon exposure to nerve growth factor and to extend neurite-like processes. Two important results were obtained. First, iron is distributed in a granular form into dopamine vesicles, found mainly in primary neurite outgrowths and distal ends. Second, thin neurite-like processes produced by differentiated cells accumulate copper, zinc, and to a minor extent lead. Overall, the high resolution imaging of single neuronal cells offers unique information to understand the role of trace metals in neurochemistry.