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Relative Quantification in Imaging of a Peptide on a Mouse Brain Tissue by Matrix-Assisted Laser Desorption Ionization

机译:通过基质辅助激光解吸电离在小鼠脑组织上的肽成像中的相对定量

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摘要

It is well-known that the number of analyte ions generated by matrix-assisted laser desorption ionization (MALDI) is not directly proportional to the analyte concentration at the irradiated spot. This is an obstacle to acquiring quantitatively meaningful maps for materials in a tissue by MALDI imaging. The problem worsens as the matrix suppression due to contaminants in the sample increases. In this work, we use a peptide as an example and show that we can overcome this problem by utilizing three guidelines derived from our recent studies on the generation of reproducible MALDI spectra. First is to acquire MALDI spectra under a temperature-controlled condition. Second is to keep the matrix suppression below an experimentally determined limit, and the third is to construct the image map using the peptide-to-matrix ion abundance ratio rather than the peptide ion abundance. The strategy works well for contaminated tissue samples and generates quantitatively meaningful maps. Also, it is demonstrated that a preposterous map can be generated when the peptide ion abundance is used in the construction of the map.
机译:众所周知,由基质辅助激光解吸电离(MALDI)生成的分析物离子的数量与照射点处的分析物浓度不直接成正比。这是通过MALDI成像获取组织中材料的定量有意义的图的障碍。随着样品中污染物的增加,基质抑制作用使问题恶化。在这项工作中,我们以肽为例,表明我们可以通过利用我们最近对可再现MALDI光谱生成的最新研究得出的三个指导方针来克服此问题。首先是在温度控制的条件下获取MALDI光谱。第二个是将基质抑制作用保持在实验确定的极限以下,第三个是使用肽与基质离子的丰度比而不是肽离子的丰度来构建图像图。该策略适用于受污染的组织样本,并生成定量有意义的图。而且,证明了当在肽图的构建中使用肽离子丰度时可以产生荒谬的图。

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