首页> 美国卫生研究院文献>Springer Open Choice >Metabolite localization by atmospheric pressure high-resolution scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging in whole-body sections and individual organs of the rove beetle Paederus riparius
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Metabolite localization by atmospheric pressure high-resolution scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging in whole-body sections and individual organs of the rove beetle Paederus riparius

机译:大气压甲虫Paederus riparius全身切片和单个器官中大气压高分辨率扫描微探针基质辅助激光解吸/电离质谱成像的代谢物定位

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摘要

Mass spectrometry imaging provides for non-targeted, label-free chemical imaging. In this study, atmospheric pressure high-resolution scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging (AP-SMALDI MSI) was used for the first time to describe the chemical distribution of the defensive compounds pederin, pseudopederin, and pederon in tissue sections (16 μm thick) of the rove beetle Paederus riparius. The whole-insect tissue section was scanned with a 20-μm step size. Mass resolution of the orbital trapping mass spectrometer was set to 100,000 at m/z 200. Additionally, organ-specific compounds were identified for brain, nerve cord, eggs, gut, ovaries, and malpighian tubules. To confirm the distribution of the specific compounds, individual organs from the insect were dissected, and MSI experiments were performed on the dissected organs. Three ganglia of the nerve cord, with a dimension of 250–500 μm, were measured with 10-μm spatial resolution. High-quality m/z images, based on high spatial resolution and high mass accuracy were generated. These features helped to assign mass spectral peaks with high confidence. Mass accuracy of the imaging experiments was <3 ppm root mean square error, and mapping of different compound classes from a single experiment was possible. This approach improved the understanding of the biochemistry of P. riparius. Concentration differences and distributions of pederin and its analogues could be visualized in the whole-insect section. Without any labeling, we assigned key lipids for specific organs to describe their location in the body and to identify morphological structures with a specificity higher than with staining or immunohistology methods.Electronic supplementary materialThe online version of this article (doi:10.1007/s00216-014-8327-1) contains supplementary material, which is available to authorized users.
机译:质谱成像可提供非目标,无标记的化学成像。在这项研究中,首次使用大气压高分辨率扫描微探针基质辅助激光解吸/电离质谱成像(AP-SMALDI MSI)来描述防御性化合物pederin,pudopederin和pederon在组织中的化学分布硬甲虫Paederus riparius的切片(16μm厚)。整个昆虫组织切片的扫描步长为20μm。轨道阱质谱仪的质量分辨率设为m / z 200时的100,000。此外,还针对大脑,神经线,卵,肠,卵巢和马尔福管确定了器官特异性化合物。为了确认特定化合物的分布,解剖了昆虫的各个器官,并在解剖的器官上进行了MSI实验。以10μm的空间分辨率测量了神经索的三个神经节,大小为250–500μm。生成了基于高空间分辨率和高质量精度的高质量m / z图像。这些功能有助于以高可信度分配质谱峰。成像实验的质量精度为<3 ppm均方根误差,可以从一个实验中绘制不同化合物类别的图谱。这种方法提高了对河滨对虾生物化学的认识。在整个昆虫区中可以观察到pederin及其类似物的浓度差异和分布。没有任何标签,我们为特定器官分配了关键脂质,以描述其在体内的位置并鉴定具有比染色或免疫组织学方法更高特异性的形态结构。电子补充材料本文的在线版本(doi:10.1007 / s00216-014) -8327-1)包含补充材料,授权用户可以使用。

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