改善了鼠标器官组织中极性和H标记代谢物的MALDI-MS成像
Siva Swapna Kasarla1, Antonia Fecke1, Karl William Smith1
1Leibniz-Institut für Analytische Wissenschaften─ISAS─e.V., Otto-Hahn-Str. 6b, Dortmund 44227, Germany.
Analytical chemistry
|May 13, 2025
概括
我们开发了一种新的六洗方法,以提高小鼠器官中极性代谢物成像的灵敏度. 这种方法增强了检测,并确保了稳定同位素标记的追踪研究的准确空间定位.
科学领域:
- 代谢学 代谢学 代谢学
- 生物医学成像技术 生物医学成像技术
- 分析化学 分析化学
背景情况:
- 使用稳定同位素标记 (SIL) 追踪成像小极性代谢物和分析它们的体内动态,对于理解生物化学途径至关重要.
- 代谢物成像方面的挑战包括低电离效率,离子抑制,以及在SIL研究中需要改进样本准备和灵敏度.
- 溶剂预处理可以提高代谢物成像灵敏度,但需要优化和空间验证以防止代谢物移位.
研究的目的:
- 优化一种简单的溶剂预处理方法,以增强对不同小鼠组织中极性代谢物的成像.
- 通过使用激光微解剖与液体染色学-协同质谱学 (LMD-LC-MS/MS) 结合,在预处理后验证代谢物的空间定位.
- 为了提高对极性代谢物的矩阵辅助激光消耗/电离质谱成像 (MALDI-MSI) 的灵敏度和稳定性.
主要方法:
- 在MALDI-MSI之前,优化"基本素"溶剂洗预处理组织截面.
- 在五个不同的小鼠器官中应用优化的方法:脏,心脏,大脑,肝脏和棕色脂肪组织.
- 使用LMD-LC-MS/MS对51种代谢物的区域特定量化进行代谢物局部化的空间验证.
主要成果:
- 基本的六洗方法显著提高了对广泛的极性和2H标记代谢物的敏感性 (高达几折).
- 该方法在各种组织类型中表现出有效性,包括脏,心脏,大脑,肝脏和棕色脂肪组织.
- LMD-LC-MS/MS分析证实了素洗后代谢物的精确空间分布,验证了MALDI-MSI的发现.
结论:
- 一个简单的"基本素"洗预处理增强了MALDI-MSI对小鼠器官中极性代谢物的敏感性.
- 开发的方法提供了一个强大的工作流程,用于对代谢物分布的敏感和空间验证的成像.
- 这种方法促进了对活体中的代谢物动态和生化途径的先进研究.
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