Three-dimensional mass spectral imaging of HeLa-M cells - sample preparation, data interpretation and visualisation
Three-dimensional mass spectral imaging of HeLa-M cells - sample preparation, data interpretation and visualisation
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DOI：
10.1002/rcm.4944
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发表时间：
2011-04-01
影响因子：
2
通讯作者：
Vickerman, John C.
中科院分区：
文献类型：
--
作者：
Fletcher, John S.;Rabbani, Sadia;Vickerman, John C.
飞行时间二次离子质谱仪(ToFSIMS)正被越来越多地应用于生物系统的研究，可以利用其化学特性和高横向分辨率成像能力。在这里，我们报告了两种细胞样品制备方法的比较，并展示了它们如何影响使用我们配备C(60)初级离子束的新型Buncher-ToF仪器(J105 3D化学成像仪)进行生物细胞三维(3D)成像的ToFSIMS分析结果。对细胞进行固定、冷冻干燥和非固定、冷冻水化分析。结论是，在分析过程中将细胞保持在非固定的冷冻-水合状态有助于减少化学再分布，产生更清晰的光谱，并改善2D和3D成像中的化学对比度。包括对数据解释的见解，我们提出了使用多变量分析技术对数据进行3D重建的方法。版权所有(C)2011 John Wiley&Sons，Ltd.
Time-of-flight secondary ion mass spectrometry (ToFSIMS) is being applied increasingly to the study of biological systems where the chemical specificity of mass spectrometry and the high lateral resolution imaging capabilities can be exploited. Here we report a comparison of two cell sample preparation methods and demonstrate how they influence the outcome of the ToFSIMS analysis for three-dimensional (3D) imaging of biological cells using our novel buncher-ToF instrument (J105 3D Chemical Imager) equipped with a C(60) primary ion beam. Cells were analysed fixed and freeze-dried and non-fixed, frozen-hydrated. It is concluded that maintaining the cells in a non-fixed frozen-hydrated state during the analysis helps reduce chemical redistribution, producing cleaner spectra and improved chemical contrast in both 2D and 3D imaging. Insights into data interpretation are included and we present methods for 3D reconstruction of the data using multivariate analysis techniques. Copyright (C) 2011 John Wiley & Sons, Ltd.