Biomolecular Imaging with a C60-SIMS/MALDI Dual Ion Source Hybrid Mass Spectrometer: Instrumentation, Matrix Enhancement, and Single Cell Analysis
Biomolecular Imaging with a C60-SIMS/MALDI Dual Ion Source Hybrid Mass Spectrometer: Instrumentation, Matrix Enhancement, and Single Cell Analysis
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DOI：
10.1007/s13361-014-0978-9
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发表时间：
2014-11-01
影响因子：
3.2
通讯作者：
Sweedler, Jonathan V.
中科院分区：
文献类型：
--
作者：
Lanni, Eric J.;Dunham, Sage J. B.;Sweedler, Jonathan V.
我们描述了一台混合MALDI/C-60-SIMS Q-TOF质谱仪和相应的样品制备方案，用于对哺乳动物脊髓、单个无脊椎动物神经元和培养的神经元网络中完整的生物分子及其片段进行成像。横向空间分辨率为10微米，进一步提高到1微米是可行的，足以解决神经元网络中细胞的生长和相互连接。这种混合仪器的高质量分辨率(913,000 FWHM)和串联质谱仪能力使其能够自信地识别细胞代谢物。升华合适的基质2，5-二羟基苯甲酸，显著增强了哺乳动物神经组织中完整的甘油磷脂离子的离子信号强度，有助于获得低丰度生物分子的高质量离子图像。这些结果表明，C-60-SIMS和MALDI质谱学的结合为需要高空间和高质量分辨率的完整生物分子成像的研究提供了特别的好处，例如单细胞、亚细胞细胞器和细胞群落的调查。
We describe a hybrid MALDI/C-60-SIMS Q-TOF mass spectrometer and corresponding sample preparation protocols to image intact biomolecules and their fragments in mammalian spinal cord, individual invertebrate neurons, and cultured neuronal networks. A lateral spatial resolution of 10 mu m was demonstrated, with further improvement feasible to 1 mu m, sufficient to resolve cell outgrowth and interconnections in neuronal networks. The high mass resolution (913,000 FWHM) and tandem mass spectrometry capability of this hybrid instrument enabled the confident identification of cellular metabolites. Sublimation of a suitable matrix, 2,5-dihydroxybenzoic acid, significantly enhanced the ion signal intensity for intact glycerophospholipid ions from mammalian nervous tissue, facilitating the acquisition of high-quality ion images for low-abundance biomolecules. These results illustrate that the combination of C-60-SIMS and MALDI mass spectrometry offers particular benefits for studies that require the imaging of intact biomolecules with high spatial and mass resolution, such as investigations of single cells, subcellular organelles, and communities of cells.