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首页> 外文会议>World biomaterials congress >Label-free 3D analysis of biological tissue with micron spatial and 240k mass resolution using a new SIMS hybrid mass analyser
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Label-free 3D analysis of biological tissue with micron spatial and 240k mass resolution using a new SIMS hybrid mass analyser

机译：使用新型SIMS混合质量分析仪进行微米级空间分辨率和240k质量分辨率的生物组织的无标记3D分析

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Introduction: Time-of-flight secondary ion mass spectrometry (TOF-SIMS) is an established, highly sensitive analytical technique for mass spectrometry (MS) imaging applications with a lateral resolution below 100 nm. Monitoring the uptake of nanoparticles, drugs or other chemicals into cells are only a few examples for the application of this label-free chemical analysis technique. Chemical information is obtained by bombarding the surface with a focused primary ion beam and analysing the generated secondary ions in a TOF mass analyzer. However in complex biological samples identification of unknown compounds can be hampered by mass interferences and a high number of possible candidates for a single mass peak. In order to overcome these limitations, the 3D nanoSIMS project is developing a revolutionary new SIMS instrument that combines the high lateral resolution and speed associated with TOF-SIMS with the high mass resolution and high mass accuracy of an orbital trapping mass analyser. The instrument is equipped with a newly developed gas cluster ion beam column allowing a lateral resolution down to the micron level. First results obtained from different biomaterials are presented here. Materials and Methods: Amiodarone-dosed NR8383 cells and a native coronal mouse brain section were analysed using either a 20 keV argon gas or a bismuth cluster primary ion beam. For MS of the generated secondary ions a new hybrid SIMS instrument was utilized, featuring a hybrid mass analyser that combines a fast TOF analyser (TOFSIMS.5, ION-TOF GmbH, Muenster, Germany) with an orbital trapping analyser (QExactive™ HF, Thermo Scientific, Bremen, Germany). Results and Discussion: fig. 1: TOF-SIMS image showing the uptake of the drug amiodarone into a single macrophage The high lateral resolution TOF-SIMS image in Fig. 1 shows the sub-cellular distribution of iodine, a moiety of the drug amiodarone (green) with respect to the nucleus of the cell (red). First gas cluster OrbitrapTM SIMS images from mouse brain sections demonstrate the simultaneous localization and identification of various compounds. On tissue, the high mass resolution (FWHM of 240 000 at m/z 200) was used to identify lipids and metabolites with sub-ppm mass accuracy. Identification of numerous lipid signals at the single cell level could be easily performed using exact mass measurement and comparison with databases. Additionally ions were selectively fragmented by tandem MS (MS/MS) in order to confirm chemical structure or help on assignment of unknown signals. Conclusion: With this unique instrument numerous applications in the field of biomaterials can be studied, including interactions between tissue and biomaterials and process control in fabrication. Laterally resolved, label-free, and non-targeted imaging of biomaterials at micron resolution is now possible.
机译：简介：飞行时间二次离子质谱仪（TOF-SIMS）是一种成熟的，高度灵敏的分析技术，适用于质谱（MS）成像应用，其横向分辨率低于100 nm。监测这种纳米颗粒，药物或其他化学物质进入细胞的摄取只是应用这种无标记化学分析技术的几个例子。通过用聚焦的一次离子束轰击表面并在TOF质量分析仪中分析产生的二次离子来获得化学信息。但是，在复杂的生物样品中，未知化合物的鉴定会受到质量干扰和单个质谱峰的大量可能候选物的阻碍。为了克服这些限制，3D nanoSIMS项目正在开发一种革命性的新型SIMS仪器，该仪器将与TOF-SIMS相关的高横向分辨率和速度与轨道阱质量分析仪的高质量分辨率和高质量精度相结合。该仪器配备了新开发的气体团簇离子束色谱柱，可实现低至微米级别的横向分辨率。本文介绍了从不同生物材料获得的第一个结果。材料和方法：使用20 keV氩气或铋团簇主离子束分析了胺碘酮给药的NR8383细胞和天然冠状小鼠脑组织。对于产生的二次离子的质谱，使用了一种新型混合SIMS仪器，其特点是混合质量分析仪将快速TOF分析仪（TOFSIMS.5，ION-TOF GmbH，德国明斯特，德国）与轨道阱分析仪（QExactive™HF， Thermo Scientific，不来梅，德国）。结果与讨论： 1：TOF-SIMS图像显示药物胺碘酮被单个巨噬细胞摄取图1中的高横向分辨率TOF-SIMS图像显示了碘的亚细胞分布，碘是药物胺碘酮相对于绿色的一部分细胞核（红色）。来自小鼠脑部的第一批气体簇OrbitrapTM SIMS图像证明了各种化合物的同时定位和鉴定。在组织上，高质量分辨率（在m / z 200时FWHM为240 000）可用于以亚ppm质量精度鉴定脂质和代谢物。使用精确的质量测量并与数据库进行比较，可以轻松地在单个细胞水平上鉴定大量脂质信号。另外，离子通过串联质谱仪（MS / MS）选择性破碎，以确认化学结构或帮助分配未知信号。结论：使用这种独特的仪器，可以研究生物材料领域的许多应用，包括组织和生物材料之间的相互作用以及制造过程中的过程控制。现在可以进行微米分辨率的生物材料的横向分辨，无标记和非靶向成像。

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