The 3D OrbiSIMS-label-free metabolic imaging with subcellular lateral resolution and high mass-resolving power
The 3D OrbiSIMS-label-free metabolic imaging with subcellular lateral resolution and high mass-resolving power
复制标题
DOI：
10.1038/nmeth.4504
复制
发表时间：
2017-12-01
期刊：
影响因子：
48
通讯作者：
Gilmore, Ian S.
中科院分区：
文献类型：
--
作者：
Passarelli, Melissa K.;Pirkl, Alexander;Gilmore, Ian S.
我们报告了一种用于无标记生物医学成像的3D OrbiSIMS仪器的开发。它结合了二次离子质谱的高空间分辨率（西姆斯;无机物质低于200 nm，生物分子低于2 mm）和Orbitrap的高质量分辨能力（m/z 200时> 240，000）。这使得外源性和内源性代谢物能够以亚细胞分辨率在3D中可视化。我们对神经递质γ-氨基丁酸、多巴胺和多巴胺的分布进行了成像，在小鼠海马体中具有很高的光谱置信度。我们还对29种磺基鞘糖脂和45种甘油磷脂的亚细胞定位进行了初步注释和绘图，并用串联质谱法确认了脂质的身份。我们使用大鼠肺泡巨噬细胞与不同浓度的药物胺碘酮孵育，证明了单细胞代谢组学分析，我们观察到磷脂种类和胆固醇的上调与胺碘酮的蓄积相关。
We report the development of a 3D OrbiSIMS instrument for label-free biomedical imaging. It combines the high spatial resolution of secondary ion mass spectrometry (SIMS; under 200 nm for inorganic species and under 2 mm for biomolecules) with the high mass-resolving power of an Orbitrap (> 240,000 at m/z 200). This allows exogenous and endogenous metabolites to be visualized in 3D with subcellular resolution. We imaged the distribution of neurotransmitters-gamma-aminobutyric acid, dopamine and serotonin-with high spectroscopic confidence in the mouse hippocampus. We also putatively annotated and mapped the subcellular localization of 29 sulfoglycosphingolipids and 45 glycerophospholipids, and we confirmed lipid identities with tandem mass spectrometry. We demonstrated single-cell metabolomic profiling using rat alveolar macrophage cells incubated with different concentrations of the drug amiodarone, and we observed that the upregulation of phospholipid species and cholesterol is correlated with the accumulation of amiodarone.