首页> 美国卫生研究院文献>Journal of Visualized Experiments : JoVE >Imaging Metals in Brain Tissue by Laser Ablation - Inductively Coupled Plasma - Mass Spectrometry (LA-ICP-MS)
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Imaging Metals in Brain Tissue by Laser Ablation - Inductively Coupled Plasma - Mass Spectrometry (LA-ICP-MS)

机译:通过激光消融-电感耦合等离子体-质谱(LA-ICP-MS)对脑组织中的金属成像

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摘要

Metals are found ubiquitously throughout an organism, with their biological role dictated by both their chemical reactivity and abundance within a specific anatomical region. Within the brain, metals have a highly compartmentalized distribution, depending on the primary function they play within the central nervous system. Imaging the spatial distribution of metals has provided unique insight into the biochemical architecture of the brain, allowing direct correlation between neuroanatomical regions and their known function with regard to metal-dependent processes. In addition, several age-related neurological disorders feature disrupted metal homeostasis, which is often confined to small regions of the brain that are otherwise difficult to analyze. Here, we describe a comprehensive method for quantitatively imaging metals in the mouse brain, using laser ablation - inductively coupled plasma - mass spectrometry (LA-ICP-MS) and specially designed image processing software. Focusing on iron, copper and zinc, which are three of the most abundant and disease-relevant metals within the brain, we describe the essential steps in sample preparation, analysis, quantitative measurements and image processing to produce maps of metal distribution within the low micrometer resolution range. This technique, applicable to any cut tissue section, is capable of demonstrating the highly variable distribution of metals within an organ or system, and can be used to identify changes in metal homeostasis and absolute levels within fine anatomical structures.
机译:金属在整个生物中无处不在,其生物学作用取决于特定解剖区域内的化学反应性和丰度。在大脑中,金属具有高度分隔的分布,这取决于它们在中枢神经系统中发挥的主要功能。对金属的空间分布进行成像已为大脑的生化结构提供了独特的见解,从而使神经解剖区域与它们在依赖金属的过程有关的已知功能之间具有直接的相关性。此外,一些与年龄有关的神经系统疾病的特征是金属稳态失调,金属稳态通常局限于大脑的小区域,否则很难分析。在这里,我们描述了一种使用激光消融-电感耦合等离子体-质谱(LA-ICP-MS)和专门设计的图像处理软件对小鼠大脑中的金属进行定量成像的综合方法。我们将重点介绍铁,铜和锌,这是大脑中三种最丰富且与疾病相关的金属,我们描述了样品制备,分析,定量测量和图像处理中的基本步骤,以生成低微米范围内的金属分布图分辨率范围。该技术适用于任何切开的组织切片,能够证明金属在器官或系统中的高度可变分布,并可用于识别金属稳态和精细解剖结构中绝对水平的变化。

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