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从单个组织部分使用 MALDI-MSI 和 LCM-microPOTS 平台增强空间蛋白质组学和代谢学

Marija Veličković¹, Le Z Day¹, Kevin J Zemaitis¹

¹Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99354, United States.

Analytical chemistry

|October 31, 2025

概括

研究人员开发了新的单组织截面 (STS) 工作流程,用于空间多组学,使其能够同时进行代谢物和蛋白质分析. 这保持了分子忠实性,并增强了组织生物学洞察力.

科学领域:

生物化学生物化学
分子生物学分子生物学
分析化学分析化学

背景情况:

基于空间分辨率质谱 (MS) 的多组学对于理解组织生物学至关重要.
目前的方法通常需要多个组织切片,由于切片间的可变性,有可能导致分子数据的相关性.
维护细胞或区域特定的分子忠实性是多组学组织分析的一个重大挑战.

研究的目的:

从单个组织部位 (STS) 开发全面的多态分析 (multiomics) 的工作流程.
通过使用不同的MS模式,从STS中实现空间代谢学和蛋白质学.
克服多部分方法的局限性,改善分子数据相关性.

主要方法:

开发了金属辅助的电绝缘基板,用于STS多组学.
使用矩阵辅助激光脱/电离-MS成像 (MALDI-MSI) 进行代谢物成像.
采用激光捕获微解剖 (LCM) 与微滴准备相结合,用于随后的蛋白质组造型.
用不同的MS仪器 (timsTOF,FTICR) 调查铜和金涂PEN基板.

主要成果:

从单个树根组织部分成功执行了非向的空间代谢和蛋白质组.
检测到>140个代谢物和6571个独特的蛋白质使用铜背PEN幻灯片与MALDI-timsTOF-MS.
分析了>170种代谢物,并使用MALDI-FTICR-MS.金PEN片识别了6542种独特的蛋白质.
取得了与独立的欧米克分析相似的结果,证明了工作流程的有效性.

结论:

开发的STS工作流程允许从单个组织部分进行高分辨率,多omics的分析.
这些方法保持了分子真实性,并为相关的多组学研究提供了新的机会.
这种方法增强了在组织内揭示复杂生物见解的能力.

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