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首页> 外文期刊>Journal of mass spectrometry: JMS >Mapping the fly Malpighian tubule lipidome by imaging mass spectrometry
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Mapping the fly Malpighian tubule lipidome by imaging mass spectrometry

机译：通过成像质谱法测定蝇膜小管脂质体

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Matrix-assisted laser/desorption ionization imaging mass spectrometry (MALDI IMS) is an analytical technique for understanding the spatial distribution of biomolecules across a sample surface. Originally employed for mammalian tissues, this technology has been adapted to study specimens as diverse as microbes and cell cultures, food such as strawberries, and invertebrates including the vinegar fly Drosophila melanogaster. As an ideal model organism, Drosophila has brought greater understanding about conserved biological processes, organism development, and diseased states and even informed management practices of agriculturally and environmentally important species. Drosophila displays anatomically separated renal (Malpighian) tubules that are the physiological equivalent to the vertebrate nephron. Insect Malpighian tubules are also responsible for pesticide detoxification. In this article, we first describe an effective workflow and sample preparation method to study the phospholipid distribution of the Malpighian tubules that initially involves the manual microdissection of the tubules in saline buffer followed by a series of washes to remove excess salt and enhances the phospholipid signals prior to matrix deposition and IMS at 25-mu m spatial resolution. We also established a complementary methodology for lipid IMS analysis of whole-body fly sections using a dual-polarity data acquisition approach at the same spatial resolution after matrix deposition by sublimation. Both procedures yield rich signal profiles from the major phospholipid classes. The reproducibility and high-quality results offered by these methodologies enable cohort studies of Drosophila through MALDI IMS.
机译：矩阵辅助激光/解吸电离成像质谱（MALDI IMS）是用于理解样品表面的生物分子的空间分布的分析技术。本技术最初用于哺乳动物组织，这项技术适用于将标本作为微生物和细胞培养物等多样化，草莓等食物，以及无脊椎动物，包括醋蝇果蝇马铃薯。作为一种理想的模型生物，果蝇对保守的生物过程，生物发展和患病状态甚至知情的经营管理实践带来了更大的了解。果蝇显示解剖学分离的肾（Malpighian）小管，这些小管是脊椎动物肾的生理学等同物。昆虫Malpighian小管也负责农药排毒。在本文中，我们首先描述了一种有效的工作流程和样品制备方法，以研究Malpighian小管的磷脂分布，其最初涉及盐水缓冲液中小管的手动微小键，然后是一系列洗涤，以除去多余的盐并增强磷脂信号在矩阵沉积之前，在25-mu m空间分辨率下。我们还建立了通过在升华矩阵沉积的相同空间分辨率下使用双极性数据采集方法来建立一种互补方法，用于全身飞行部分。这两种程序都从主要磷脂等级产生了丰富的信号型材。这些方法提供的再现性和高质量的结果使通过MALDI IMS队列的果蝇研究。

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